March 2024
Dysthyroidism in patients with psychiatric disorders in Morocco
Soukaina Stati 1, 6, Ahmed Kharbach 2, 3, Mohamed Amine Baba 2, 4, Aya Kaoutar Rhaouti 1, 7, Aouatef Khallouk 1, 7, Majdouline Obtel 3, 5, Fouad Laboudi 1, 6, Abderrazzak Ouanass 1, 6
1 Department of Psychiatry, Ibn Sina University Hospital, Rabat, 2 Department of Public Health, High Institute of Nursing Professions and Health Technics, Agadir, 3 Department of Biostatistics, Faculty of Medicine and Pharmacy, Mohamed V University, Rabat, 4 Department of Public Health, Faculty of Medicine and Pharmacy, Ibn Zohr University, Agadir, 5 Department of Social Medicine Public Health, Faculty of Medicine and Pharmacy, Mohamed V University, Rabat, 6 Department of Psychiatry, Faculty of Medicine and Pharmacy, Mohamed V University, Rabat, 7 Departmentof Psychiatry , Faculty of Medicine and Pharmacy, Mohamed V University, Rabat, Morocco
DOI: 10.4328/ACAM.21353 Received: 2022-08-09 Accepted: 2022-09-21 Published Online: 2024-01-22 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):141-144
Corresponding Author: Mohamed Amine Baba, Department of Public Health, Laboratory of Health Sciences Research, Faculty of Medicine and Pharmacy, Ibn Zohr University, Agadir, Morocco. E-mail: babamedamine2@gmail.com P: +21 263 344 25 54 Corresponding Author ORCID ID: https://orcid.org/0000-0002-6660-9527
Aim: In this study, we aimed to explore the association between dysthyroidism and psychiatric disorders and to verify whether there are sociodemographic or family variables that can determine the likelihood of this comorbidity.
Material and Methods: A descriptive, analytical, cross-sectional and retrospective study, involved patients aged between 18 and 65 years, followed for thyroid pathologies comorbid with psychiatric disorders.
Results: The participants were women with the median age of 52 years, 69.4% of patients presented a mood disorder, 30.6% had another psychiatric disorder, 6.1% presented a clinical goiter with a normal biological assessment, 8.2% presented hyperthyroidism and 85.7% presented hypothyroidism. Age, family situation, obstetrical, family and surgical ATCD, therapeutic compliance of psychiatric and thyroid disease and pharmacological treatment of psychiatric disorders are variables that present a statistically significant difference with a P <0.05 between the two groups.
Discussion: Dysthyroidism and especially hypothyroidism is associated more with mood disorders than other psychiatric disorders in hospitalized patients.
Keywords: Mood Disorder, Dysthyroidism, Woman, Morocco
Introduction
The occurrence of mental disorders during endocrine pathologies has been established for more than a century. Conversely, psychological trauma may play a role in triggering certain endocrinopathies.
The association between thyroid function and psychiatric disorders, especially mood disorders, was recognized more than 200 years ago [1, 2]. Patients with mood disorders, including bipolar disorders and depressive disorders have a high prevalence of somatic comorbidity compared to the general population [3, 4]. Hypothyroidism is relatively common in these patients [4], this comorbidity would be correlated with a high risk of relapses and resistance to treatment, and suicide attempts [5, 6]. It is well established that the use of psychiatric drugs (eg lithium) increases the risk of thyroid dysfunction [3, 4, 7]
The role of thyroid dysfunction in depression is recognized [8]. Depressive symptoms often occur in patients with thyroid disorders and, conversely, patients with depression may have various thyroid abnormalities, including high or low levels of thyroid hormones [1].
Recent studies have confirmed a strong association between thyroid dysfunction in patients with mood disorders [9]. For example, Ojha et al reported that 21.4% of depressed patients had abnormal thyroid function, 11.4% met criteria for subclinical hypothyroidism, 5.7% for subclinical hyperthyroidism, and 4.3% for overt hypothyroidism [10]. Regarding the relationship between thyroid dysfunction and bipolar disorder, a recent study by Ying Zhao et al shows that early onset of bipolar disorder is correlated with higher rates of hypothyroidism [11].
In addition, thyroid dysfunction could have a significant influence on the cognition and emotion of those affected [1]. However, although previous studies have revealed associations between mood disorders and thyroid dysfunction, the roles of thyroid hormones, including free triiodothyronine (FT3), free thyroxine (FT4), (TSH), anti-thyroglobulin (TgAb) and anti-thyroid peroxidase antibodies (TPOAb) in the pathological mechanisms of mood disorders were not yet clear [1, 12].
The main objective of this study was to explore the association between dysthyroidism and psychiatric disorders, to confirm this association especially in patients with mood disorders and to verify whether there is a sociodemographic or family profile that may determine the likelihood of this comorbidity.
Material and Methods
A descriptive, cross-sectional, retrospective study made at the Ar-Razi Sale Psychiatric University Hospital in Morocco, from January 2014 to January 2020.
Data Collection
This study focused on patients whose age varied between 18 and 65 years. The female sex was chosen in our study due to a high frequency of this comorbidity in women compared to men during our research at the level of the Ar-Razi Psychiatric University hospital for patients followed for thyroid pathologies comorbid with psychiatric disorders whose diagnosis was based on the DSM 5 diagnostic criteria. The records of male patients, female patients under the age of 18 and records with missing data were excluded. The clinical data was collected using a pre-established exploitation sheet with socio-demographic data (age, origin, profession, socio-economic level, marital status), clinical data (personal and family history, circumstances of consultation, revealing psychiatric manifestations). The collection of clinical data was carried out with respect for the anonymity and confidentiality of patients with medical records from the archives of the Ar-Razi Psychiatric University Hospital.
Statistical analysis
Statistical analysis was carried out using JAMOVI software for Windows 2016. Qualitative variables were presented as frequencies and percentages, quantitative variables were presented in mean standard deviation (SD) or median (interquartile range, IQR). The Chi-square (x2) test or Fisher’s exact test were carried out according to their particular application conditions to identify the differences in the proportions of the categorical variables between the two groups (Group 1: dysthyroidism comorbid with a disorders, Group 2: dysthyroidism comorbid with another psychiatric disorder other than a mood disorder). In addition, multivariate logistic regression analyzes are used to identify risk factors. Independent variables presenting a statistically significant value with P < 0.05 between the two groups were taken into account in multivariate logistic regression.
Results
A total of 49 participants who met the study criteria were included in the study. The average age was 52 [41.59] years, more than half (53.1%) had a secondary education level, 55.1% were married, 95.9 % lived with a family, 55.1% did not have a profession. the majority were without antecedents 73.5%, and 28.6% without a child. More than half (69.4%) have a mood disorder, 30.6% had another psychiatric disorder (not a mood disorder). The majority (85.7%) had hypothyroidism, 6.1% had a clinical goiter with a normal biological assessment, 8.2% present hyperthyroidism.
Psychiatric disorder and dysthyroidism
By comparing the two groups and using the Chi-square (x2) test or Fisher’s exact test, we found that there is a statistically significant difference with a P <0.05 for age, family situation, obstetrical history, family and surgical, therapeutic compliance of psychiatric and thyroid disease and pharmacological treatment of psychiatric disorders see (Table 1).
Multinomial logistic regression
Using multivariate logistic regression and adjusting for confounding factors, concluding that being old, single, with surgical ATCDs and poor treatment compliance are risk factors for developing a mood disorder or even (Table 2).
Discussion
According to the results of our study, the participants were women with a median age of 52 [41.59] years, more than half (53.1%) had a secondary education level, the majority had no background 73.5%, 69.4% of patients presented a mood disorder, 30.6% had other psychiatric disorder (without mood disorder), 6.1% presented a clinical goiter with a normal biological assessment, 8.2% presented a hyperthyroidism and 85.7% presented a hypothyroidism.
Age, family situation, obstetrical, family and surgical history, therapeutic compliance with psychiatric and thyroid disease and pharmacological treatment of psychiatric disorders are variables that present a statistically significant difference with a P <0.05 between the two groups.
Risk factors for developing a mood disorder were older age, being single, a history of surgery and poor adherence to treatment for psychiatric disorders and dysthyroidism.
The female sex was chosen in our study due to a high frequency of this comorbidity in women compared to men. Several studies confirm our observation and found that women were associated with an increased risk of hypothyroidism comorbid with mood disorder [11].The same results were obtained in a retrospective cohort study, which included 13,017 subjects, and which revealed that the rate of depression was twice as high in women with TSH levels above 2.3 μIU/Ml [13].
In the present study the median age was 52 [41.59] years, and our results consistent with a recent study which showing that the prevalence of hypothyroidism in the subgroup with an age below 50 years was 8.53%, which was lower than that of patients over 50 years old, which was (10.8%) [11]. It is also it is consistent with another previous study that age was a risk factor for hypothyroidism [14].
Our results show that the association between dysthyroidism and psychiatric disorders is frequent, especially with mood disorders in 69.4% of patients, these results are consistent with the results of a recent study that shows an association between early onset of a psychiatric disorder and hypothyroidism, which may be present specifically in mood disorders, rather than non-specifically in other serious mental illnesses, as this association has not been observed in schizophrenia [11].
According to our results and the results of previous studies, there is a relationship between thyroid dysfunction and mood disorders. Thyroid hormones have multiple effects on body homeostasis and metabolism [15]. It is therefore not surprising that there are documented relationships between thyroid status and mood disorders. Specifically, thyroid disorders may increase the risk of mood disorders and the rate of depression is high in people with hypothyroidism [16].
Thyroid hormones exert a profound influence on behavior and appear capable of modulating the phenotypic expression of major mood disorders. Indeed, a recent study found that there is evidence that triiodothyronine (LT3) can accelerate antidepressant response to antidepressants, and that LT3 can also increase the response to antidepressants in depression resistant to medical treatment [17], always in the same study, adjunctive treatment with supraphysiological doses of levothyroxine (LT4) has shown its effectiveness in several studies, including rapid cycling and prophylaxis-resistant bipolar disorder, and in refractory acute unipolar or bipolar depression [17]. Another explanation for this comorbidity is related to immune system abnormalities that are seen in both thyroid and depressive disorders, but although the most common cause of thyroid disorders is autoimmune disease, dysregulated immune function, and not autoimmunity, is more clearly documented in depression [18, 19].
In addition to a high frequency of this comorbidity, resistance to antidepressant treatment is also documented. Bruce M. Cohen reported that depression in patients with high TSH levels is less severe but more resistant to treatment and he observed a weaker antidepressant response, especially to selective serotonin reuptake inhibitors, in patients with major depression and higher TSH levels, with no correlation of response to serum T 3 or T 4 levels [20].
The complex relationship between selective serotonin reuptake inhibitors and thyroid function remains clarified, on the one hand, hypothyroidism alters responses to SSRIs [20], and on the other hand, SSRIs cause a decrease in T4 levels, of free T4 and triiodothyronine (T3) without an appropriate compensatory response in TSH [21], one of the hypotheses to answer this question is that major depression is associated with a subtle chronobiological dysregulation of the hypothalamic-pituitary-thyroid axis ( HPT) [22].
There are several limitations to this study. First, no causal relationship could be established due to the cross-sectional design of this study. Future studies should use a longitudinal study design to better explore the causal relationship of these variables in patients with this comorbidity.
Second, the minimal number of patients was 49. Future research should include a wider range of patients with this comorbidity and both females and males.
Third, some confounding factors that may play an important role in mood disorders, and future research should incorporate more confounding factors to better elucidate the pathological mechanisms underlying the association between thyroid dysfunction and psychiatric disorders. Finally, we did not have a healthy control group in this study. Therefore, our results in this study should be considered preliminary, which merits replication in future studies before we can draw any conclusions.
Conclusion
Dysthyroidism and especially hypothyroidism are associated more with mood disorders than with other psychiatric disorders, and clinicians must look for this association, so as not to miss hypothyroidism which, without treatment, will aggravate the clinical picture of a disorder from mood to resistance to pharmacological treatment and increased prevalence of suicide.
Longitudinal cohort studies are needed to explore the clinical significance and potential mechanism of this association.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and human rights statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. No animal or human studies were carried out by the authors for this article.
Funding: None
Conflict of interest
None of the authors received any type of financial support that could be considered potential conflict of interest regarding the manuscript or its submission.
References
1. Hage MP, Azar ST. The link between thyroid function and depression. J Thyroid Res. 2012. DOI: 10.1155/2012/590648.
2. Medici M, Direk N, Visser WE, Korevaar TI, Hofman A, Visser T, et al. Thyroid function within the normal range and the risk of depression: a population-based cohort study. J Clin Endocrinol Metab. 2014;99(4):1213-19.
3. Correll C U, Detraux J, De Lepeleire J, De Hert M. Effects of antipsychotics, antidepressants and mood stabilizers on risk for physical diseases in people with schizophrenia, depression and bipolar disorder. World Psychiatry. 2015;14(2):119-36.
4. Firth J, Rosenbaum S, Galletly C, Siddiqi N, Stubbs B, Killackey E, Protecting physical health in people with mental illness–Authors’ reply. The Lancet Psychiatry. 2019;6(11):890-1.
5. Amann BL, Radua J, Wunsch C, König B, Simhandl C. Psychiatric and physical comorbidities and their impact on the course of bipolar disorder: a prospective, naturalistic 4-year follow-up study. Bipolar Disorders. 2017;19(3):225-34.
6. Cole DP, Thase ME, Mallinger AG, Soares JC, Luther JF, Kupfer DJ, et al. Slower treatment response in bipolar depression predicted by lower pretreatment thyroid function. American Journal of Psychiatry. 2002;159(1):116-21.
7. Hayes JF, Marston L, Walters K, Geddes JR, King M, Osborn DP. Adverse renal, endocrine, hepatic, and metabolic events during maintenance mood stabilizer treatment for bipolar disorder: a population-based cohort study. PLoS medicine. 2016;13(8).
8. Shen Y, Wu F, Zhou Y, Ma Y, Huang X, Ning Y, et al. Association of thyroid dysfunction with suicide attempts in first-episode and drug naive patients with major depressive disorder. Journal of affective disorders. 2019;259(3):180-5.
9. Charnsil C, Pilakanta S. Prevalence Of Thyroid Dysfunction And Its Relationship With The Severity Of Major Depressive Disorder (Mdd). ASEAN Journal of Psychiatry. 2017;18(1):1-5.
10. Ojha SP, Dhungana S, Chapagain M, Tulachan P. Association of thyroid dysfunction with depression in a teaching hospital. Journal of Nepal Health Research Council. 2013;11(23):30-4.
11. Zhao Y, Wang N, Chen J, Guan Q, Yuan X, Shen Y, et al. Association between early-onset affective disorders and hypothyroidism in a larger number of psychiatric drug-free patients. Journal of Affective Disorders. 2022;299:31-6.
12. Euesden J, Danese A, Lewis CM, Maughan BA bidirectional relationship between depression and the autoimmune disorders–New perspectives from the National Child Development Study. PloS One. 2017;12(3). DOI: 10.1371/journal.pone.0173015
13. Kim EY, Kim SH, Rhee SJ, Huh I, Ha K, Kim J, et al. Relationship between thyroid-stimulating hormone levels and risk of depression among the general population with normal free T4 levels. Psychoneuroendocrinology. 2015;58:114-19.
14. Leng O, Razvi S. Hypothyroidism in the older population. Thyroid Research. 2019;12(1):1-10.
15. Delitala AP, Scuteri A, Doria C. Thyroid hormone diseases and osteoporosis. Journal of clinical medicine. 2020;9(4):1034.
16. Samuels MH. Psychiatric and cognitive manifestations of hypothyroidism. Current opinion in endocrinology, diabetes, and obesity. 2014;21(5):377.
17. Bauer M, Whybrow PC. Role of thyroid hormone therapy in depressive disorders. Journal of Endocrinological Investigation. 2021;44(11):2341-47.
18. Ellul P, Gasnier M, Trebossen V, Gaillard R. Immune activation and deficit in neurotransmitters synthesis in treatment resistant depression: about a case of hashimoto encephalopathy. Clinical Psychopharmacology and Neuroscience. 2020;18(3):463-6.
19. Stein DJ, Naudé PJ, Berk M. Stress, depression, and inflammation: molecular and microglial mechanisms. Biological psychiatry. 2018;83(1):5-6.
20. Cohen BM, Sommer BR, Vuckovic A. Antidepressant-resistant depression in patients with comorbid subclinical hypothyroidism or high-normal TSH levels. American Journal of Psychiatry. 2018;175(7):598-604.
21. Caye A, Pilz LK, Maia AL, Hidalgo MP, Furukawa TA, Kieling C. The impact of selective serotonin reuptake inhibitors on the thyroid function among patients with major depressive disorder: a systematic review and meta-analysis. European Neuropsychopharmacology. 2020;33:139-45.
22. Duval F. Thyroid hormone treatment of mood disorders. Current Treatment Options in Psychiatry. 2018;5(4):363-76.
Download attachments: 10.4328.ACAM.21353
Soukaina Stati, Mohamed Amine Baba, Ahmed Kharbach, Aya Kaoutar Rhaouti, Aouatef Khallouk, Majdouline Obtel, Fouad Laboudi, Abderrazzak Ouanass. Dysthyroidism in patients with psychiatric disorders in Morocco. Ann Clin Anal Med 2024;15(3):141-144
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
Retrospective evaluation of lumbar puncture analysis in the pediatric intensive care unit
Melis Öztürk Aykaç 1, Mustafa Törehan Aslan 2, Nedim Samancı 3
1 Department of Pediatrics, 2 Department of Pediatrics, Division of Neonatology, 3 Department of Pediatrics, Division of Pediatric Intensive Care, Faculty of Medicine, Tekirdağ Namık University, Tekirdağ, Turkey
DOI: 10.4328/ACAM.21832 Received: 2023-07-26 Accepted: 2023-08-28 Published Online: 2023-12-29 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):145-148
Corresponding Author: Nedim Samancı, Department of Pediatrics, Division of Pediatric Intensive Care, Faculty of Medicine, Tekirdağ Namık University, Tekirdağ, Turkey. E-mail: samancitr@yahoo.com P: +90 532 295 71 98 Corresponding Author ORCID ID: https://orcid.org/0000-0002-3947-3492
This study was approved by the Ethics Committee of Tekirdag Namik Kemal University Non-Interventional Clinical Research Ethics Committee (Date: 2021-03-30, No: 2021.66.03.06)
Aim: Meningitis is a central nervous system infectious disease with inflammation of the leptomeninges and subarachnoid space. There is still a severe mortality and morbidity from meningitis. Therefore, it is essential to make an early diagnosis and start empirical treatment for the causative agent as soon as possible.
Material and Methods: Seventy patients diagnosed with meningitis in our hospital were included in our study. The anamnesis, physical examination, laboratory results, and treatment of the patients were retrospectively reviewed.
Results: The most common complaint of our patients was fever, with 84.3%. Meningeal irritation finding was positive in 86% of patients aged two years and older, while this rate was 22.4% in patients under two years of age. According to the meningeal irritation findings, a statistically significant difference was found in terms of neutrophil, lymphocyte, and platelet values (p<0.05). Neutrophil levels of those with positive signs of meningeal irritation were significantly higher than in those without symptoms. Lymphocyte and platelet values of those with positive signs of meningeal irritation were substantially lower than in those without. The median values were 11.5 g/dL for hemoglobin, 11220/uL for leukocytes, 321500/uL for platelets, 25.9 mg/dl for CRP, 29 mm/h for ESR, 61.2 mg/dL for CSF glucose, 37.4 mg/dL for CSF protein, and 100 for cell count.
Discussion: In 4 of our patients, the CSF culture detected the causative microorganism. In 3 of the culture-negative patients, the causative pathogen was detected in CSF PCR. The most frequently preferred empirical antimicrobial treatment initiated for patients is the combination of vancomycin and a third-generation cephalosporin.
Keywords: Meningitis, Fever, Meningeal Irritation Signs, Cerebrospinal Fluid
Introduction
Meningitis is a central nervous system infectious disease that describes the inflammation of the meninges and the cerebrospinal fluid in the subarachnoid space resulting in severe mortality and morbidity [1, 2]. Bacteria, viruses, and other infectious agents can cause meningitis [1]. Clinical findings and morphological, chemical, and microbiological examination of the cerebrospinal fluid can reveal the underlying cause. The main goal in treating meningitis is to diagnose and start treatment as soon as possible, to prevent or minimize the complications that may develop [3, 4]. This research was carried out in the pediatric intensive care unit of our hospital to evaluate the clinical findings, blood tests, chemical and microbiological examinations in the cerebrospinal fluid and to examine the demographic characteristics, application complaints, examination findings, and test results of the patients diagnosed with meningitis, the compatibility of the biochemical and microbiological parameters obtained from these results, and the contribution of all these results to the diagnosis [1, 5].
Material and Methods
Demographic characteristics, anamnesis, and physical examination findings, biochemical and microbiological data of 70 patients aged one month to 18 years who underwent lumbar puncture with the diagnosis of meningitis in Tekirdağ Namık Kemal University Hospital, Department of Pediatrics, Pediatric Intensive Care Unit were included in the study by retrospectively examining. Ethical approval was obtained from Tekirdag Namik Kemal University Non-Interventional Clinical Research Ethics Committee (Study protocol number: 2021.66.03.06, Date: 30.03.2021).
Statistical analyses
Statistical analyses were performed using a package program called SPSS (IBM SPSS Statistics 24). Frequency tables and descriptive statistics were used to interpret the findings. Parametric methods were used for measurement values suitable for a normal distribution. Using parametric methods, the “Independent Sample-t” test (t-table value) method was used to compare the measurement values of two independent groups. Non-parametric methods were used for the measurement values that did not conform to the normal distribution. By non-parametric methods, the “Mann-Whitney U” test (Z-table value) method was used to compare the measurement values of two independent groups. The Spearman correlation coefficient was used to examine the relationships of two quantitative variables that do not have a normal distribution. “Pearson-χ2” crosstabs were used to explore the relationships between two qualitative variables.
Results
It was determined that the mean age of the patients was 6.11±5.33 (years), and 27 (38.6%) were in the ≤1 age group. It was determined that 40 patients (57.2%) applied in the spring and summer months; 41 (58.6%) were male, 29 (41.4%) were female, and 5 (7.1%) had ventriculoperitoneal shunts (Table 1). A statistically significant difference was found in admission complaints regarding age (years) and head and neck pain status (Z=-6.431; p=0.0001). A statistically significant difference was found in age (years) according to MIB status at the first examination (Z=-5.829; p=0.0001). It was observed that the age of those with MIB was significantly higher than that of those without. A statistically significant correlation was found between the patient group under two years of age and the MIB status (χ2=25.677; p=0.0001). A statistically significant correlation was found between the patient group under two years of age and anterior fontanel status (χ2=35,758; p=0.0001). It was determined that the anterior fontanel of 17 individuals (63%) under the age oftwo years was open. Moreover, it was determined that those in the <2 age group were predominantly open (bomb and pulsatile), while those in the age group over two years old were all closed. Thirty-one of the patients (44.3%) had very intense leukocytes in the CSF culture, 64 (91.4%) had PMNL as the dominant cell, 66 (94.3%) had no growth in the CSF culture, four patients (5.7%) had change in their culture, two patients (50.0%) had change in CSF culture, pneumococcal growth was found in 3 (4.2%) of meningitis. There was no statistically significant difference in terms of leukocytes, neutrophils, lymphocytes, platelets, and C-Reactive Protein (CRP) according to the presence of fever in complaints at admission (p>0.05). A statistically significant difference was found in erythrocyte sedimentation rate (ESR) values according to the presence of fever in complaints at admission (Z=-2.442; p=0.015). It was observed that the ESR values in those who had an uproar in the application complaint were significantly higher than in those who did not have a fever. There was no statistically significant difference in leukocytes, neutrophils, CRP, and ESR values according to the presence of head and neck pain in the presenting complaint (p>0.05).
A statistically significant difference was found in lymphocyte and platelet values according to head and neck pain status (p<0.05). There was no statistically significant difference in the admission complaints regarding nausea, vomiting, sleepiness, leukocytes, neutrophils, platelets, CRP, and ESR values (p>0.05). There was no statistically significant difference in leukocytes, neutrophils, lymphocytes, CRP, and ESR according to seizure status at admission (p>0.05). However, a statistically significant difference was found in platelet values (t=-2.135; p=0.036). Platelet values of those with seizure complaints were significantly higher than in those without seizures. At the first examination, there was no statistically significant difference in leukocytes, CRP, and ESR according to the meningeal irritation findings in the patients (p>0.05). According to the meningeal irritation findings, a statistically significant difference was found in terms of neutrophil, lymphocyte, and platelet values (p<0.05). Neutrophil levels in those with positive signs of meningeal irritation were significantly higher than in those without symptoms of meningeal irritation. A positive, weak, and statistically significant correlation was found between the patients’ leukocyte values and CRP values (r=0.399; p=0.001). As CRP increases, leukocytes will increase. Likewise, it was observed that leukocyte values decreased as CRP decreased. A positive, weak, statistically significant relationship was found between ESR and CRP (r=0.440; p=0.0001). As CRP increases, ESR will increase. When the distribution of the patients’ biochemical and microscopic examination results was examined, the median cell number was f00/mm3, the median CSF protein was 61.2 mg/dl, and the median CRP was 25.9 mg/dl. The median leukocyte count was 11220, which is detailed in Table 2.
Discussion
Childhood meningitis has been one of the most feared infectious diseases since ancient times [1]. Acute bacterial meningitis, which continues to cause death and disability in children, is a neurological emergency, especially in developing countries with weak immunization studies [6]. Despite advances in antimicrobial treatments, the rate of development of acute bacterial meningitis in some developing countries varies between 16-32% and is still very high [7, 8].
Most of our study’s patients (38.6%) were one year old or younger. We think that this result is due to the immaturity of the immune response and the lumbar puncture to be performed primarily when no fever focus can be found in a patient younger than one year of age and the vaccine doses have not been completed yet. It has been stated that the only risk factor is age under 1 year, and the mortality rate under 1 year of age is high; therefore, this age group should be closely monitored. The same study emphasized that the reasons behind the uncertain seasonal bacterial meningitis patterns should be investigated further. Considering the application dates of the 70 patients in our study, according to the seasonal distribution we made, the total number of patients admitted in the spring and summer months (57.2%) is higher than the number of patients admitted in the autumn and winter months (42.8%). However, no significant difference was observed between the distributions when the seasons were evaluated alone.
The most common finding in bacterial meningitis is fever [8]. When the symptoms of 70 patients in our study were examined, the most common symptom was fever in 59 patients (84.3%). The most common symptom after turmoil is head and neck pain in 34 (48.6%) patients. A statistically significant difference was found in age (years) according to head and neck pain status in admission complaints. Those with head and neck pain are significantly older than those without. Considering both the relationship of anterior fontanel status with age and the requirement that children must be above a certain age to define head and neck pain as a complaint, it is customary to be statistically significant.
A statistically significant difference was found in admission complaints regarding age (years) and head and neck pain status (Z=-6.431; p=0.0001). It was observed that the ages of those with head and neck pain were significantly higher than those without. A statistically significant difference was found in age (years) according to MIB status at the first examination (Z=-5.829; p=0.0001). It was observed that the ages of those with MIB were significantly higher than those without.
In the study by Michos et al., in which they examined 506 patients with aseptic meningitis in Greece, the median cell number in the CSF was found to be 201/mm3, and PMNL was the dominant cell in 58.3% of the patients [9]. PMNL predominates in the CSF in approximately 90% of patients with bacterial meningitis. PMNL predominates in the early period in 20-75% of cases with viral meningitis [10]. In the CSF cell analysis of 70 patients in our study, the median cell number was 100 per mm3, and in the CSF analysis of 64 (94.3%) of the patients, PMNL was dominant when cell typing was performed. S.pneumoniae growth was detected in 2 of 4 patients with development.
In our study, five patients (7.1%) had ventriculoperitoneal shunts, and all meningitis patients with VP shunts were younger than one year old. Enterococci were found in CSF culture in 1 patient with VP shunt, and Methicillin-Resistant Staphylococcus hominis growth was detected in CSF culture in 1 patient. However, since the number of patients with VP shunt is limited, it is difficult to make a judgment.
The diagnosis of bacterial meningitis is confirmed by microbiological testing of CSF, which includes cytochemical analysis, cell count, microscopy, and culture [11, 12]. However, antibiotics before the lumbar puncture procedure may reduce the culture yield and cause diagnostic uncertainty [12, 13].
In our study, when the symptoms and signs were compared with some biochemical parameters, ESR values in patients with fever were statistically significantly higher than in those without fever. No statistically significant difference was found between the patients with and without fever for values of other leukocytes, neutrophils, lymphocytes, and CRP. Lymphocyte and platelet values in those with head and neck pain were statistically significantly lower than in those without head and neck pain. It was observed that neutrophil values in patients with seizures were statistically significantly higher than in those without seizures. Neutrophil values in those with meningeal irritation findings were statistically significantly higher than in those without, and lymphocyte and platelet values were lower.
Conclusion
The mean age of the patients was 6.11 ± 5.33 (years), and 37.2% were one year old or younger. When patients’ admission symptoms and physical examination findings are examined, the most common symptom is fever, with a rate of 84.3%. ESR values in patients with fever at admission are statistically significantly higher than in those without. No statistically significant difference was found between the patients with and without fever for other values of leukocytes, neutrophils, lymphocytes, and CRP. The most common symptom after turmoil is head and neck pain, which is seen in 48.6%, and a statistically significant difference was found in age (years) according to head and neck pain status in admission complaints (Z=-6.431; p=0.0001). A statistically significant difference was found in lymphocyte and platelet values according to head and neck pain status (p<0.05). The lymphocyte and platelet values in those with head and neck pain were significantly lower than in those without head and neck pain.
The meningeal irritation findings found a statistically significant difference in neutrophil, lymphocyte, and platelet values (p<0.05).
Growth was observed in the CSF culture of four patients, and S. pneumoniae growth was detected in 2 of them. In the other two patients with development, VP shunt was found. Enterococcus growth was seen in one of the CSF cultures, and Methicillin-Resistant Staphylococcus hominis growth was found in the CSF culture of the other patient. Five of the patients included in our study had VP shunt, and all of the patients with VP shunt were under one year old. In patients in whom the CSF PCR was studied, no growth was observed in the CSF culture of 3 patients, and N. meningitidis was found in one patient, Hib in one, and Parechovirus in the other in the CSF PCR results. The causative pathogen was detected by CSF culture and PCR methods in only seven of 70 patients in our study. Although the etiology cannot be determined, the patients are those whose meningitis is suspected by anamnesis, physical examination, and laboratory findings, and the diagnosis of meningitis is confirmed by the CSF’s cell count and the CSF’s biochemical analysis. For this reason, especially unexplained fever, headache, vomiting, and positive signs of meningeal irritation on physical examination, the presence of a bulging and pulsatile fontanelle in a child with open anterior fontanelle should bring meningitis to mind in the diagnosis, and especially if there are no contraindications, the presence of meningitis should be considered. If suspected, acyclovir treatment for HSV should be started.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: None
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Hasbun R. Progress and Challenges in Bacterial Meningitis: A Review. Jama. 2022;328(21):2147-54.
2. Van Hentenryck M, Schroeder AR, McCulloh RJ, Stave CD, Wang ME. Duration of Antibiotic Therapy for Bacterial Meningitis in Young Infants: A Systematic Review. Pediatrics. 2022;150(5):e2022057510 DOI: 10.1542/peds.2022-057510.
3. Silva GD, Guedes BF, Junqueira IR, Gomes HR, Vidal JE. Diagnostic and therapeutic approach to chronic meningitis in Brazil: A narrative review. Arq Neuropsiquiatr. 2022;80(11):1167-77.
4. Mijovic H, Sadarangani M. To LP or not to LP? Identifying the Etiology of Pediatric Meningitis. Pediatr Infect Dis J. 2019;38(6S Suppl. 1):S39-42.
5. de Almeida SM, Castoldi JR, Riechi SC. Comparison between cerebrospinal fluid biomarkers for differential diagnosis of acute meningitis. Diagnosis (Berl). 2023; 10(3):298-308 DOI: 10.1515/dx-2023-0013.
6. Kuti BP, Bello EO, Jegede TO, Olubosede O. Epidemiological, clinical and prognostic profile of childhood acute bacterial meningitis in a resource poor setting. J Neurosci Rural Pract. 2015;6(4):549-57.
7. Jarousha AM, Afifi AA. Epidemiology and Risk Factors Associated with Developing Bacterial Meningitis among Children in Gaza Strip. Iran J Public Health. 2014;43(9):1176-83.
8. Nickerson JW, Attaran A, Westerberg BD, Curtis S, Overton S, Mayer P. Fatal Bacterial Meningitis Possibly Associated with Substandard Ceftriaxone–Uganda, 2013. MMWR Morb Mortal Wkly Rep. 2016;64(50-51):1375-7.
9. Michos AG, Syriopoulou VP, Hadjichristodoulou C, Daikos GL, Lagona E, Douridas P, et al. Aseptic meningitis in children: Analysis of 506 cases. PloS one. 2007;2(7):e674.
10. Kanra G, Ceyhan M, Kara A. Meningitis II: Clinical findings. Çocuk Sağlığı ve Hastalıkları Dergisi/ Journal of Child Health and Diseases. 2003;46(2):128-38.
11. Polage CR, Cohen SH. State-of-the-Art Microbiologic Testing for Community-Acquired Meningitis and Encephalitis. J Clin Microbiol. 2016;54(5):1197-202.
12. Khumalo J, Nicol M, Hardie D, Muloiwa R, Mteshana P, Bamford C. Diagnostic accuracy of two multiplex real-time polymerase chain reaction assays for the diagnosis of meningitis in children in a resource-limited setting. PloS one. 2017;12(3):e0173948.
13. Abdelrahim NA, Mohammed N, Evander M, Ahlm C, Fadl-Elmula IM. Viral meningitis in Sudanese children: Differentiation, etiology and review of literature. Medicine. 2022;101(46):e31588.
Download attachments: 10.4328.ACAM.21832
Melis Öztürk Aykaç, Mustafa Törehan Aslan, Nedim Samancı. Retrospective evaluation of lumbar puncture analysis in the pediatric intensive care unit. Ann Clin Anal Med 2024;15(3):145-148
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
Effects of high-level noise on the consumption of anesthetic agents during total intravenous anesthesia
Ayse Mizrak 1, Elzem Sen 1, Lutfiye Pirbudak 1, İbrahim Erkutlu 2, Rauf Gul 1, Betul Kocamer Simsek 3
1 Department of Anesthesiology and Reanimation, Faculty of Medicine, Gaziantep University, 2 Department of Neurosurgery, Faculty of Medicine, Gaziantep University, 3 Department of Anesthesiology and Reanimation, Faculty of Medicine, Sanko University, Gaziantep, Turkey
DOI: 10.4328/ACAM.21917 Received: 2023-09-04 Accepted: 2024-01-08 Published Online: 2024-01-23 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):149-153
Corresponding Author: Elzem Sen, Department of Anesthesiology and Reanimation, Faculty of Medicine, Gaziantep University, 27310, Sahinbey, Gaziantep, Turkey. E-mail: drelzemsen@gmail.com P: +90 532 784 21 51 Corresponding Author ORCID ID: https://orcid.org/0000-0003-3001-7324
This study was approved by the Ethics Committee of Gaziantep University (Date: 2009-04-09, No: 2009/120)
Aim: The purpose of this study is to investigate the effects of high-level noise on propofol and remifentanyl consumption in patients who undergo cholecystectomy surgery under total intravenous anaesthesia (TIVA) during operation and on the satisfaction of patients and physicians.
Material and Methods: In this study, 90 patients who would undergo cholecystectomy surgery were randomized in 3 groups. It was planned as a double blind, prospective, randomized clinical trial. Propofol and remifentanyl infusion was started to obtain BIS values between 40-60 for all patients. The patients in Group N (n=30) were subjected to noise of normal operating room. The patients in Group S (n=30) were subjected to noise between 80-85 Db with headphone and for patients in Group Q subjection to noise of the operating room was prevented by covering the ears with headphones (n=30). Intraoperative total remifentanyl and propofol consumption of the patients, postoperative patient and physician satisfaction, postoperative side effects were recorded.
Results: Total remifentanyl and propofol used during surgery was significantly lower in Group Q than in Group N and Group S (p=0,0001, p=0,04). Postoperative patient satisfaction in Group Q was statistically higher than in Group N and Group S (p=0.001). Surgeon satisfaction in Group Q was also higher than in Group N and Group S (p=0.01).
Discussion: High noise level for patients who undergo cholecystectomy surgery under TIVA increases total remifentanyl and propofol consumption during operation. In addition, it decreases patient and surgeon satisfaction.
Keywords: Noise, Propofol Consumption, Remifentanil Consumption
Introduction
Noise is defined as an unpleasant and unwanted sound. Environmental noise pollution is regarded as a general stressor, increasing mental stress, the development of cerebral cardio- vascular disease, and the risk of hearing loss [1]. Previous studies reported that the normal noise level in operating room was between 51- 79 dB and maximum noise level was between 80- 119 dB [1,2]. A quality improvement project underlined that noise can no longer be ignored as an unchangeable certainty in the operating room. Although patients were generally satisfied with their care, anesthetists perceived that excessive noise in the operating room negatively affected their ability to perform in the operating room. Noise is partly ingrained in operating room culture, but noise reduction can be achieved through training [3].
The role of music in the treatment of preoperative anxiety in adult patients was investigated. Researchers concluded that listening music before surgery had anxiolytic effects [4]. And in parallel with this, it was reported that music decreased sedative and analgesic requirements and reduced BIS score. Kühlmann et al. [5] reported that music provided a clinically meaningful reduction in pain and has been shown to reduce postoperative analgesic use. Capenetto et al. [6] stated that the most important effect of music applied to surgical patients is related to its positive effect on their psychological aspects. Ligree et al. [7] showed that noise-cancellation headphones can reduce anxiety and improve sedation scores.
But, in another study they reported that adding music during surgery does not provide any additional benefit when noise reduction technology is used. [8]. Therefore, we planned our study to make the answers of these questions clearer. In our study, we investigated the effect of noise on propofol and remifentanyl consumption and, anxiety in patients applied TIVA and sedation level in the patient and the surgeon during perioperative period.
Material and Methods
The study was conducted after obtaining the written informed consent from the patients. Ninety patients with ASA (American Society of Anesthesiologists) I-II whose ages were between 18-65 who would underwent cholecystectomy surgery under TIVA (Total Intravenous Anaesthesia) were included in the study. The study was planned as a randomized, double blind, prospective clinical study. Written consents of the patients were received after a detailed explanation was made on the previous day before the study, about anesthesia method to be performed. The work presented has been performed in accordance with the most recent version of the Helsinki Declaration.
The patients with midazolam or remifentanyl sensitivity, who had history of psychiatric drug usage recently, who had active respiratory tract infection and ear infection, who were addict of alcohol, narcotics or drugs, who had renal, cardiac or liver dysfunction, who were pregnant and did not want to be included to the study, were not included in the study. Audible level of the patients before surgery were tested audiometrically and the patients with auditory deficit were excluded from the study.
Preoperative sedation was not applied to any of the patients. Peripheral venous catheter was inserted, non-invasive blood pressure, peripheric oxygen saturation (SPO2), ECG (electrocardiogram) and BIS (Bispektral Index) were monitored after all the patients were entered in the operating room. Than demografhic values were evaluated with modified Observer’s Assessment of Alertness/Sedation Score (OAA/S) 5 minutes before operation and at the 0th. minute. OAA/S: 0 = no response to painful stimulant; 1 = no response to delicate shaking and nudging; 2 = response only to delicate shaking and nudging; 3 = response only when called with noisy and repeatitive voice; 4 = lethargic response when called with normal voice; 5 = ready response when his/her name is called with normal voice [9]. Emergence agitation score (behaviour score) (1= sleeping, 2= awake, calm, 3= irritable, 4= inconsolable crying, 5= severe restlessness, disorientation) [10] and basal anxiety score were evaluated and also heart rate (HR) and mean arterial pressure (MAP) was recorded. The application of anesthesia was performed by a single assistant and the evaluation was performed by another individual blinded to the group of the patient. The patients were randomized according to the computer ranks.
The patients were intubated 2-4 minutes after anethesia induction was applied with standard doses of rocuronium bromide (Esmeron® vial 10 mg.ml-1, Organon, Oss, Holland) 0.5 mg/kg and propofol (Propofol 1% Fresenius®, 10 mg.ml-1, Fresenius Kabi AB, Uppsala, Sweden) 2-2.5 mg/kg, and they were connected to anesthesia machine. Maintenance of anesthesia was continued as TIVA by applying remifentanyl (Ultiva® 2 mg.ml-1, Glaxo Smith Kline, S.p.A, Italy) and propofol infusion with different perfusors (Braun Infusomat, Melsungen, Germany). Therefore, 0.01-0.1 µ/kg/min remifentanyl infusion and 6-10 mg/kg/hour propofol infusion were performed for BIS value to be in the range 40-60. Mechanical ventilation was performed with 50% O2 and 50% air. BIS measuring device (Aspect Medical Systems, Natick, MA) was used for monitorization. The patients were randomized into 3 groups according to the order in the computer. The patients in high noise group [Group S (n=30)] were subjected to noise in the environment together with induction (Noise of the alarm was increased, everybody spoke loudly, and music was listened to on the radio) and they were made to listen to traffic noise at the level of 80-85 dB with headphone. The sources of noise are different types of equipment; the conversations between the workers and the alarms of different equipment [11-13].
The ears were covered with headphones tightly in Group Q (silence group). Silence of the environment was maintained as far as possible. In the patient group where normal room noise was maintained [Group N (n=30)], the ears of the patients were left open so that they could be sensitive to the ambient noise. TIVA was terminated after surgical intervention was completed.
HR, MAP, and BIS values were recorded during the intraoperative period at minutes 0, 5, 10, 15, 30, 45, and 60 and HR, MAP, sedation score and anxiety scores were recorded at minutes 0 and 5 after they were awakened. Postoperative total remifentanyl and propofol consumption of the patients; operation duration, recovery duration from anesthesia (time passed from extubation until verbal communication is established in postoperative observation room), patient and surgeon satisfaction scores (0= not satisfied, 1= slightly satisfied, 2= satisfied, 3= very satisfied); side effects like postoperative nausea, vomiting, tachycardia, bradycardia (HR<50/dk), hypertension, hypotension (MAP<60 mmHg), coughing were also evaluated and recorded. Aneshesia and data collection were performed in each group by a person who had no information about the other group.
Statistical Analysis
SPSS (Statistical Package for Social Sciences) for Windows 16.0 statistics program was used in the assessment of the parameters studied. The demographic features of each group were compared by means of variance analysis. In order to analyze and compare the between-groups parametric data (comparison of MAP, HR, recovery time, duration of surgery, total remifentanil consumption and total propofol consumption) one-way ANOVA was used. The anxiety, sedation and satisfaction scores of the groups were compared by means of the Kruskal-Wallis test. The nausea-vomiting, bradycardia, tachycardia, hypertension, hypotension and coughing were compared with χ2 test. All the data were expressed as mean ± standard deviation or percentage % or median (minimum-maximum). The statistical significance level was determined to be meaningful at p<0.05. We did not calculate the sample size. However, for purposes of the power calculation, a 25% increase in consumption of propofol and remifentanyl was considered to be significant.
Ethical Approval
This study was approved by the Ethics Commitee of Gaziantep University (Date: 2009-04-09, No: 2009/120)
Results
There was not any significant difference between the groups with regards to demographic data, recovery time from anesthesia and surgery (p>0.05) (Table 1). Total remifentanyl consumption determined during operation in Group Q was statistically significantly lower than Group N and Group S (p=0.0001) (Table 1). Total propofol consumption determined during operation in Group Q was statistically significantly lower than Group N and Group S (p=0.04) (Table 1). There was no statistical difference between groups with regard to preoperative and postoperative anxiety scores and sedation scores (p>0.05) (Table 2).
The postoperative patient satisfaction score in Group Q was statistically significantly higher than Group N and Group S (p=0.001) (Table 2). Postoperative surgeon satisfaction score in Group Q was statistically significantly higher than Group N and Group S (p=0.01) (Table 2).
A statistically significant difference was not observed with regards to postoperative complications like nausea, vomiting, hypertension, hypotension tachycardia, bradycardia and coughing (p>0.05) (Table 3).
Discussion
We investigated the effects of high-level noise in patients who underwent cholecystectomy surgery under TIVA on propofol and remifentanyl consumed during operation and postoperative anxiety and sedation levels. We observed that high-level noise increased the total propofol and remifentanyl consumed during operation. Besides, we determined that patient and surgeon satisfaction was significantly higher in the silence group.
Bispectral index (BIS) is a useful monitor for the evaluation of sedation, hypnosis and loss of consciousness and for the decrease of drug consumption, for the prevention of awareness and for providing short recovery periods [14, 15]. TIVA ensures fast induction, balanced maintenance of anaesthesia and also decreases side effects like nausea, vomiting, and shivering. The best combination in TIVA is generally obtained with remifentanyl and propofol which is hypnotic and analgesic [16].
Many studies performed before, showed that the noise level in the hospitals is far above the recommended noise level [6]. WHO (World Health Organisation) recommends a noise level of up to 30 dB in operating rooms but in reality, the noise level in operating rooms is higher than this value [2]. The studies performed before stated that the average noise level in operating rooms was between 51-79 dB and the maximum level was between 80-119 dB [2]. In our operating room the average noise is 65 dB. In our study, we used traffic noise between 80-85 dB together with ambient noise in the noise group both to generate adequate and not to cause damage to the patients .
It is known that audio warnings organize stress responses [17, 18]. In some studies, it was stated that music had reduction effect on stress during surgery and on anxiety before surgery [6]. But, different from this, noise may cause harmful hormonal changes associated with stress response and secondary effects such as hypertension on circulatory physiology [1]. Noise is also a stress source for employees and may disturb the concentration and the mental activity of the employees. It may impair the personal performance of the surgeon during the operation. Disturbance of mental activity is one of the most important reasons for medical error and side effects and this subject should be taken seriously [17]. In other words, noise may be a detrimental factor not only for the patients but also for the employees of the hospital. [19]. Noise causes physiological reactions such as increased blood pressure and long-term exposure can cause cardiovascular, metabolic and mental health disorders [20]. As noise is an important discomforting factor, healthcare workers should be protected from excessive noise.
At the end of our study, we determined that noise increased intraoperative propofol and remifentanyl consumption. In most studies performed with patients who underwent surgery under anesthesia, it was stated that music had effects in the direction that decreased the consumption of sedatives and analgesics [21-23]. Perioperative music may reduce the need for postoperative opioids and intraoperative sedative medications. Therefore, as higher opioid dosage is associated with an increased risk of side effects and chronic opioid use, perioperative music may potentially improve patient outcomes and reduce medical costs. Although no side effects were observed, the use of perioperative music appears to be safe and patient-friendly, given the reported high patient satisfaction [23]. Giordano et al. [24] reported that preoperative music therapy could be an alternative to intravenous midazolam when aiming to promote a preoperative and postoperative state of anxiolysis and sedation in stomatology surgery, even if no differences were found in terms of the surgery-related stress response according to physiological and hormonal determinations.
These studies suggest that while music is theoretically expected to decrease stress hormones in patients under general anaesthesia, noise might increase the consumption of sedatives and analgesics by increasing stress hormones. An increase in stress hormones related to noise may induce hypertension and this shall increase the requirement for propofol and remifentanyl. The increase in stress hormones might be a reason for anxiety in both the patients who recover from anaesthesia and the staffs working in the same environment. Surgeon satisfaction and postoperative patient satisfaction were the highest without noise pollution.
Conclusion
We observed that noise increased consumption of propofol and remifentanyl in patients who underwent cholecystectomy surgery under TIVA. The consumption was determined as a minimum in silence group. Furthemore, patient and surgeon satisfaction was also higher in the silence group.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: None
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Fu VX, Oomens Pim, Merkus N, Jeekel J. The Perception and Attitude Toward Noise and Music in the Operating Room: A Systematic Review. J Surg Res. 2021;263(1):193-206.
2. Mcleod R, Myint-Wilks L, Davies SE, Elhassan HA. The Impact of Noise in the Operating Theatre: A review of the evidence . Ann R Coll Surg Engl. 2021;103(2):83-7.
3. Yu CV, Foglia J, Yen P, Montemurro T, Schwarz SKW, MacDonell S. Noise in the operating room during induction of anesthesia: Impact of a quality improvement initiative. Can J Anaesth. 2022;69(4):494-503.
4. Petot T, Bouscaren N, Maillard O, Huiart L, Boukerrou M, Reynaud D. Comparing the effects of self-selected music versus predetermined music on patient anxiety prior to gynaecological surgery: A study protocol for a randomised controlled trial. Trials. 2019;20(1):20.
5. Kühlmann AYR, de Rooij A, Kroese LF, van Dijk M, Hunink MGM, Jeekel J. Meta-analysis evaluating music interventions for anxiety and pain in surgery. Br J Surg 2018;105(7):773–83.
6. Capenetto P, LaMattina G, Quattropani MC. Music therapy and psychological-clinical impact in surgery: A systematic review. Health Psychol Res. 2022;10(4):38615.
7. Ligree N, Nanda S, Morwal S, Garg K. Effect of binaural beat music and noise cancelling headphones on intraoperative anxiety in patients undergoing spinal anaesthesia- A randomised controlled study. Indian J Anaesth. 2023;67(7):590-4.
8. Tran BW, Nowrouz MY, Dhillon SK, Xie KK, Breslin KM, Golladay GJ. The impact of music and noise-cancellation on sedation requirements during total knee replacement: A randomized controlled trial. Geriatr Orthop Surg Rehabil. 2020;11(1):1-7.
9. Schüttler J, Eisenried A, Lerch M, Fechner J, Jeleazcov C, Ihmsen H. Pharmacokinetics and pharmacodynamics of remimazolam (CNS 7056) after continuous infusion in healthy male volunteers: Part I. Pharmacokinetics and clinical pharmacodynamics. Anesthesiology. 2020;132(4):636-51.
10. Menser C, Smith H. Emergence agitation and delirium: Considerations for epidemiology and routine monitoring in pediatric patients. Local Reg Anesth. 2020;13(1):73-83.
11. Hampton T, Everett S, Sharma S, Krishnan M. Noise in the operating theatre. Br J Surg. 2021;108(5):203-4.
12. Giv MD, Sani KG, Alizadeh M, Valinejadi A, Majdabadi HA. Evaluation of noise pollution level in the operating rooms of hospitals: A study in Iran. Interv Med Appl Sci. 2017;9(2):61-6.
13. Hamad F, Moacdieh NM, Banat R, Lakissian Z, Al-Qaisi S, Zaytoun G, et al. Perceptions on music and noise in the operating room : A cross-sectional study. Int J Occup Saf Ergon. 2022;28(4):2168-72.
14. Oliveira CR, Bernardo WM, Nunes VM. Benefit of general anesthesia monitored by bispectral index compared with monitoring guided only by clinical parameters. Systematic review and meta-analysis. Braz J Anesthesiol. 2017;67(1):72-84.
15. Li Z, Cai J, Li J, Xu X, Zheng L. Comparative evaluation of the bispectral index (BIS) and BISpro during propofol anesthesia. J Int Med Res. 2021;49(4):3000605211001705.
16. Ramirez MF, Gan TJ. Total intravenous anaesthesia versus inhalation anesthesia: How do outcomes compare? Curr Opin Anaesthesiol. 2023:36(4):399-406.
17. Ukegjini K, Kastiunig T, Widmann B, Warschkow R, Steffen T. Impact of intraoperative noise measurement on the surgeon stress and patient outcomes. A single-center clinical trial with 664 patients. Surgery. 2020;167(5):843-851.
18. Baltin CT, Wilhelm H, Wittland M, Hoelscher AH, Stippel D, Astvatsatourov A. Noise patterns in visceral surgical procedures: analysis of second -by-second dBA data of 599 procedures over the course of one year. Sci Rep. 2020;10(1):3030.
19. Lee A, Torkamani-Azar M, Zheng B, Bednarik R. Unpacking the broad landscape of intraoperative stressors for clinical personnel: A mixed-methods systematic review. J Multidiscip Healthc. 2023;16(1):1953-77.
20. Salazar MR. Hypertension, a linchpin between environmental noise exposure and the development of cardiovascular disease? J Clin Hypertens (Greenwich). 2023;25(2):165-7.
21. Tajbakhsh A, Salimi S, Daftarian N, Abtahi D. Effect of music during general anesthesia on anesthetic consumption during vitrectomy surgery. Adv Biomed Res.2023;12:59.
22. Singh P, Arya A, Singh MK, Prakash R, Khan MP. Effect of spiritual music on old-age patients undergoing lower limb surgery under spinal anesthesia. Anesth Essays Res. 2022;16(2):208-212
23. Fu VX, Oomens P, Klimek M, Verhofstad MHJ, Jeekel J. The effect of perioperative music on medication requirement and hospital length of stay: A meta-analysis. Ann Surg. 2020;272(6):961-972.
24. You S, Xu F, Wu Y, Qin S, Shu B, Chen Y, Chen Y, Huang H, Duan G. Effect of noise isolation using noise-cancelling headphones during laparoscopic surgery for postoperative pain reduction: A randomized clinical trial. J Clin Anesth. 2024;92:111286.
Download attachments: 10.4328.ACAM.21917
Ayse Mizrak, Elzem Sen, Lutfiye Pirbudak, İbrahim Erkutlu, Rauf Gul, Betul Kocamer Simsek. Effects of high-level noise on the consumption of anesthetic agents during total intravenous anesthesia. Ann Clin Anal Med 2024;15(3):149-153
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
Risk factors for neonatal respiratory distress : Moroccan retrospective study
Latifa Mochhoury 1, Fatima Dahmanı 2, Touria Essayagh 1, Ikram Marc 1, Amina Barkat 3
1 Department of science Laboratory of Health Sciences and Technologies, Higher institute of Health Sciences, Hassan First University, Settat, 2 Department of Hematology, Faculty of Medicine and Pharmacy Research Team in hemalology, Mohammed V. University, Rabat, 3 Department of Neonatology Research Team in Maternal and Child Health and Nutrition, Faculty of Medicine and Pharmacy, Mohammed V University, Rabat, Morocco
DOI: 10.4328/ACAM.21998 Received: 2023-09-27 Accepted: 2023-11-06 Published Online: 2023-12-06 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):154-159
Corresponding Author: Latifa Mochhoury, Department of science Laboratory of Health Sciences and Technologies, Higher institute of Health Sciences, Hassan First University, Settat Morocco. E-mail: latifa.mochhoury@uhp.ac.ma P: +212 620 303 5 82 Corresponding Author ORCID ID: https://orcid.org/0000-0002-5019-2296
This study was approved by the Ethics Committee of CERB of the Faculty of Medicine and Pharmacy (Date: 2021-24-02, No: C64/20)
Aim: The aim of the study was to determine the risk factors for neonatal respiratory distress.
Material and Methods: This retrospective analytical study was conducted at the National Reference Center for Neonatology and Nutrition at Children’s Hospital, University Hospital Centre IbnSina of Rabat from January 1, 2021 to December 31, 2022.
During the study period, 630 newborns who met the inclusion criteria were included in 2 groups: neonates with respiratory distress (n=421) and neonates without respiratory distress (n=209). The identification of risk factors was carried out using bivariate as well as multivariate analyses
Results: 630 births were collected during this period Neonatal respiratory distress was multifactorial. Statistical analysis could reveal mostly maternal anemia (OR = 18.10; CI95 (7.5 -43.55); p ˂ 0.05), diabetes (OR = 3.65; CI 95 (1.98-6.72); p = 0.001), caesarean section (OR = 4.23; CI 95 (1.54-11. 59); p = 0.001), prematurity (OR = 2.45; CI 95 (1.41-4.26); p =0.01).
Discussion: Screening, management, and reduction of neonatal respiratory distress remain a crucial challenge, requiring early coordination between pediatricians and obstetricians to obtain reliable data and identify newborns at risk.
Keywords: Neonatal Respiratory Distress, Maternal Anemia, Diabetes, Prematurity
Introduction
The birth of newborns is associated with intrinsic and extrinsic alterations that ensure the transition from the intrauterine to the extrauterine environment particularly at the pulmonary and cardiovascular levels. At times Generally, the newborn adapts naturally to extra-uterine conditions.However, in some cases, this naturaladaptation fails, leading to cardiac arrest associated with hypoxia and severe neurological damage, requiring immediate and rapid management..[1].
Neonatal respiratory distress is caused by a deficit in oxygen supply leading to a change in the physiologic state of the newborn. This incident compromises the vital prognosis of the infant in particular by alternating phases of hypoxia and acidosis. Additionally, neonatal respiratory distress is often the product of a number of vectors such as hyaline membrane disease, late amniotic fluid resorption, prematurity, fetal asphyxia and neonatal infections. In underdeveloped countries, neonatal respiratory distress is more common given lower socioeconomic conditions and more prevalent risk factors leading to perinatal complications such as poor or absent infrastructure [2] that threaten maternal and fetal survival.
In addition to the lack of research, there is a lack of information on this syndrome in Morocco, so we aimed this study to provide valuable data in order to help identify the frequency of respiratory distress and the risk factors associated with this major public health problem in newborns in the neonatal unit of Children’s Hospital of Rabat.
In Morocco, Neonatal Respiratory Distress Syndrome (NRDS) is recognized as a major public health problem and a constant concern in neonatology units due to its frequency and severity.
Our results highlight the importance of specific risk factors in the development and severity of NRDS and can be used to inform evidence-based NRDS management protocols in the Neonatal Intensive Care Unit (NICU), develop strategic planning for obstetric management, and hopefully set the basis for further epidemiological studies.
Material and Methods
Study Population and Design
This retrospective analytical study was conducted at the National Reference Center for Neonatology and Nutrition at Children’s Hospital of Rabat from 1 January 2021 to December 31, 2022. The study concerned hospitalized newborns for neonatal respiratory distress. As a tertiary hospital, the neonatal unit of Children’s Hospital of Rabat is one of the main neonatal units in Morocco. Through consecutive convenience sampling, we included all complete and eligible files of newborns admitted within the early neonatal period (first seven days of life). Data were extracted from the files into a case record form.
Study variables
Data collection was carried out by a documentary technique consisting of studying the medical records of each neonate. All data were entered using an information sheet containing the following sections:
• Maternal and obstetrical characteristics
• Characteristics of the newborn
• Evaluation of the patient
• Dependent variable: neonatal respiratory distress
• Independent variables:
‣ Socio-demographic characteristics: age, residence, marital status, educational and socioeconomic status of the mother, gestational age and area of origin
‣ Obstetrics-related factors: gravidity, parity, current mode of delivery.
‣ Newborn characteristics (sex, birth weight, Apgar score, Silverman score, and time to respiratory distress> or <3 hours.
‣ Immediate resuscitation at birth.
‣ Amniotic rupture.
‣ Maternal pathologies during pregnancy (gestational diabetes, pre-eclampsia, goiter, asthma and anemia).
‣ Evaluation of the severity is based on a Silverman score, which is composed of inspiratory and expiratory categories of movements.
The scale of the Silverman score ranges from 0 to 2:
• Neonatal moderate respiratory distress corresponding to Silverman ≤4
• Intense Neonatal Respiratory Distress corresponding to Silverman between 4 – 6
• Very intense Neonatal Respiratory Distress corresponding to Silverman >6
Inclusion Criteria:
Symptomatic or asymptomatic neonates hospitalized for clinical management of neonatal respiratory distress were included.
Exclusion Criteria:
– All newborns with congenital malformations
– Incomplete data sheets
– Respiratory distress of surgical origin.
Definitions of used terms:
• neonatal respiratory distress is defined by the presence of at least one of the following elements: abnormal respiratory rate (tachypnea > 60 breaths/min; bradypnea < 30 breaths/min; respiratory pauses, or apnea) or signs of labored breathing (expiratory grunting, nasal flaring, intercostal recessions, xyphoid recessions or thoracoabdominal asynchrony) ) = • Silverman’s score = A score greater than 7 indicates that the baby is in respiratory failure.
•Delay of care at birth: refers to the time it takes to seek care after the onset of labor that is longer than 1 hour.
•Primimarous: a woman pregnant for the first time
•Multiparous: a woman who has had multiple births
•Premature rupture of membranes (PROM) is a rupture (breaking open) of the membranes (amniotic sac) before labor begins.
Data Analysis
•The descriptive analysis of the variables was based primarily on class size ; proportions. Mean and standard deviations were used as measures of central tendency and dispersion. Qualitative variables were compared using the chi-square test or Fisher’s exact test.The Kolmogorov-Smirnov test was used for the study of the distribution of the variables. Pearson’s correlation test was performed to understand the relationships between quantitative variables. Comparisons of means of quantitative variables for different classes of qualitative variables were performed using the Student’s t-test for independent samples. After verification of the different conditions of the test for all statistical tests, p <0.05 was considered significant.
Ethical Approval
• This study was approved by the Biomedical Research Ethics Committee CERB of the Faculty of Medicine and Pharmacy on (Date: 2021-24-02, No: N/R : File n° C64/20).
• This study is a part of the research project “Near Miss Neonatal in Morocco“
Results
We included 630 newborns, among whom 421 had neonatal respiratory distress symptoms NRDS.
Table 1 shows that the median maternal age was 35 years (Q1-Q3: [22; 36]). The distribution according to educational level was dominated by illiteracy and secondary education with respective percentages of 48.2% and 44.2%, p<0.05. On the other hand, the university level was only 7.6% of the population. The mean gestational age of the newborns was 36.8 ± 8.86 gestational weeks; socioeconomic status was low in 54.2% of the cases and medium in only 45.8%, p<0.05.
Table 2 indicates that among 421 neonates with neonatal respiratory distress, 190 were female and 231 were male, with a sex ratio of 1.21; the sex of the neonate had no effect on neonatal respiratory distress in our study (p=0.62), the mean birth weight was 3000g (Q1-Q3: [2100; 4050]).
The cesarean section prevailed in 80.5% of cases, with a significant statistical difference p<0.05; early rupture of membranes >12 hours was 84.1% and aspect of amniotic fluid (55.1%) p<0.0
The main maternal and neonatal pathologies associated with respiratory distress were anemia (p<0.05), pre-eclampsia (p=0.02), maternal infection (p<0.05) and gestational diabetes (p<0.05). The main identified causes of respiratory distress were transitory tachypnea (16.5%), maternal-fetal infection (19.6 %), hyaline membrane disease (30. 1%) and prematurity (54%).
Multiple regression statistical analysis primarily incriminated anemia (OR = 18.10; 95 CI (7.5 -43.55); p <0.05), diabetes (OR = 3.65; 95 CI (1.98-6.72); p = 0.001), cesarean section (OR = 4.23; 95 CI (1.54-11.59); p = 0. 001), prematurity (OR = 2.45; 95 CI (1.41-4.26); p = 0.01), appearance of amniotic fluid (OR = 27.9; 95 CI (13.46-55.34); p < 0.005); premature rupture of membranes (OR = 5.40; 95 CI (2.58-11.29); p < 0.05), and early resuscitation at birth (OR = 30.95; 95 CI (13.65-70.13); p < 0.05), (Table 3).
Discussion
In this study, we registered 420 newborns with respiratory distress (66.6%); several risk factors for respiratory distress have been identified.
Age was a determinant of neonatal respiratory distress in our study. We found that the median age of our patients was 35 years (Q1-Q3: [22, 36]). Indeed, maternal age between 30 and 40 is well known to be associated with morbidity and neonatal respiratory distress, which may be explained by the increased risk of chronic diseases such as diabetes and hypertension at this age [3, 4].
In our sample, there is a relationship between low maternal education and neonatal respiratory distress. Indeed, low level of education seems to be a factor limiting perinatal health care and the use of medical services due to the lack of awareness and information.
Many studies have shown that the rate of fetal respiratory distress is inversely proportional to the mother’s level of education [6]. This could be due to a better diet, better hygiene and the greater use of health services by educated women.
In Morocco, lack of access to emergency neonatal care is widespread among women living in rural and isolated communities, therefore women are far from medical facilities.
54.9% were male with a sex ratio of 1.21; Our results are not significant regarding the association between neonatal respiratory distress and gender. That’s contradictory,because male gender is a risk factor for respiratory distress due to the frenative effect of androgens on surfactant synthesis, which delays lung maturation in males compared to females [7]. Conversely, female fetuses produce surfactant earlier and have more developed lung parenchyma and airways [8, 9].
The average weight was 3000g (Q1-Q3: [2100; 4050]); neonatal respiratory distress was higher in newborns weighing less than 2500g; thus, 54.4% of newborns were hypothrophic, 13.5% eutrophic and 32.1% macrosomia. Our study is in agreement with study [10], which explains that hypotrophy is secondary to chronic hypoxia induced by hypertension and to nutrient delivery to the fetus due to decreased uteroplacental perfusion. Thus, Doppler velocimetry analysis of umbilical artery blood flow correlates with increased respiratory complications in neonates of hypertensive mothers [11].
Oxygen therapy is systematically used in all cases of respiratory distress and in all newborns (full term or premature). Our study reports that a majority (94.4%) of newborns were not resuscitated at birth (oxygen therapy; artificial ventilation).
This confirms that the management at birth of the newborns in our sample presents major difficulties. Indeed, newborns come from peripheral health centers and district hospitals; therefore, referrals are made by poor quality and non-medical transport. Alamneh et al. identified that the main risk factors for birth asphyxia were fetal distress, instrumental delivery, low birth weight, non-cephalic presentation, preterm, prolonged labor, co-morbidity during pregnancy and Meconium-stained amniotic fluid (MSAF). Most of these factors are preventable through holistic care for pregnancy, labor and delivery. Therefore, to reduce neonatal mortality associated with birth asphyxia, attention should be paid to holistic pregnancy care services, labor and delivery care. Furthermore, intervention strategies aimed at reducing birth asphyxia should target the identified factors [12].
We can say that neonatal respiratory distress is closely related to cesarean section, which is consistent with the study that showed a ten times higher incidence of neonatal airways . In theory, the delay in resorption of alveolar fluid when adapting to the extrauterine environment during a programmed caesarean section induces hormonal imbalance of catecholamines and a defect of maturation of the fetal lungs leading to neonatal respiratory distress [13, 14].
Theoretically, the delay in resorption of alveolar fluid during adaptation to the extra-uterine environment during a programmed caesarean section induces a hormonal imbalance in the catecholamines released and a defect in the maturation of the fetal lungs [15].
The relationship between maternal diabetes mellitus (DM) and neonatal respiratory distress syndrome (RDS) has long been recognized.
Our results are in line with meta-analysis suggesting that maternal DM is linked to an increased risk of neonatal RDS, especially when diabetes is not well controlled, maternal hyperglycemia causes a disorder of carbohydrate metabolism in the developing embryo with alterations in various metabolic pathways that create oxidative stress [16].
Our result is in line with studies [17], which showed that serious complications lead to intensive care unit admission, causing a serious burden on healthcare units. Paying more attention to the healthcare needs of pregnant women can help identify preeclampsia earlier and minimize the complications associated with it.
High blood pressure and high proteinuria are two main characteristics of preeclamptic patients, affecting the organs of the mother and fetus [18].
Our results confirm that anemia was a global problem with serious consequences for mothers and their babies. Even though anemia in pregnancy is readily treatable, data from several studies show an association between maternal anemia and severe adverse maternal and perinatal outcomes [19, 20].
We are confident that our findings are a robust demonstration of an independent link between severe anemia and SDR. Although our results do not explain a direct maternal cause of anemia ; iron and folic acid deficienciescan be incriminated, probably due to both inadequate nutrition and low socio-economic status. Prevention and treatment of anemia during pregnancy and post-partum should remain a global public health and research priority.
Conclusion
Neonatal respiratory distress is multifactorial and requires special attention. This study highlighted the seriousness of this syndrome. It also showed that the main risk factors are prematurity, cesarean section, gestational diabetes, anemia and pre-eclampsia.
Consequently, a thorough understanding of these factors enables us to implement preventive strategies.
We can propose the following recommendations:
– Strengthening the technical and human resources of the neonatology department, referral hospitals and provincial hospitals. – Training in neonatal resuscitation techniques during classes.
– Early medical referral of severe cases is essential.
– Improving the health status of mothers and newborns
Limitations of this work:
Our study was retrospective and our database was limited and many parameters are missing. We conducted this study during COVID-19, thus, access to different unit was not free.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: None
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Zec SL, Selmanovic K, Andrijic NL, Kadic A, Zecevic L, Zunic L. Evaluation of drug treatment of bronchopneumonia at the pediatric clinic in Sarajevo. Med Arch. 2016;70(3):177-81.
2. Nascimento-Carvalho AC, Ruuskanen O, Nascimento-Carvalho CM. Wheezing independently predicts viral infection in children with community-acquired pneumonia. Pediatr Pulmonol. 2019;54(7):1022-8.
3. Abdelgabar SA, Babikr WG, Elhussein AB, Hummeida ME. Risk of respiratory morbidity in term infants delivered by elective cesarean delivery. Int J Med Sci Public Health. 2016;5:973-7.
4. Wyckoff MH, Aziz K, Escobedo MB, Kapadia VS, Kattwinkel J, Perlman JM, et al. Neonatal resuscitation: 2015 American Heart Association Guidelines update for cardiopulmonary resuscitation and emergency cardiovascular care. Pediatrics. 2015;136(Suppl.2):S196-218.
5. Timofeev J, et al. Obstetric complications. neonatal morbidity, and indications for cesarean delivery by maternal age. Obstet Gynecol. 2013;122(6):1184-95.
6. Beaujouan É, Reimondos A, Gray E, Evans A, Sobotka T. Declining realisation of reproductive intentions with age. Hum Reprod. 2019;34(10):1906-14.
7. Bongen E, et al. Sex Differences in the Blood Transcriptome Identify Robust Changes in Immune Cell Proportions with Aging and Influenza Infection. Cell Rep. 2019;29(7):1961-73.e4.
8. Silveyra P, Fuentes N, Rodriguez Bauza DE. Sex and gender differences in lung disease. Adv Exp Med Biol. 2021;1304:227-58.
9. Shibata M, Ogawa K, Kanazawa S, Kawasaki M, Morisaki N, Mito A, et al. Association of maternal birth weight with the risk of low birth weight and small- for-gestational-age in offspring: a prospective single-center cohort study. PLoS One. 2021;16(5):e0251734.
10. Barreto CTG, Tavares FG, Theme-Filha M, Farias YN, Pantoja LN, Cardoso AM. Low birthweight, prematurity, and intrauterine growth restriction: results from the baseline data of the first indigenous birth cohort in Brazil (Guarani Birth Cohort). BMC Pregnancy Childbirth. 2020;20(1):781.
11. Mehari M, Maeruf H, Robles CC, Woldemariam S, Adhena T, Mulugeta M, et al.
Advanced maternal age pregnancy and its adverse obstetrical and perinatal outcomes in Ayder comprehensive specialized hospital, northern Ethiopia, 2017: A comparative cross-sectional study. BMC Pregnancy Childbirth. 2020;20(1):60.
12. Alamneh YM, Negesse A, Aynalem YA, Shiferaw WS, Gedefew M, Tilahun M, et al. Risk Factors of Birth Asphyxia among Newborns at Debre Markos Comprehensive Specialized Referral Hospital, Northwest Ethiopia: Unmatched Case-Control Study.Ethiop J Health Sci. 2022;32(3):513-22.
13. Tefera M, Assefa N, Mengistie B, Abrham A, Teji K, Worku T. Elective Cesarean Section on Term Pregnancies Has a High Risk for Neonatal Respiratory Morbidity in Developed Countries: A Systematic Review and Meta-Analysis. Front. Pediatr. 2020;8:286.
14. Saddi YIA, Baythoon MB, Tahir AG. The timing of elective caesarean deliveries and early neonatal respiratory morbidity in term neonates. J Faculty Med. 2018;60:38-42.
15. Raveloharimino H. Fiangoa F. Ratianarison F. Rabesandratana HN. Epidemio- clinical profile of respiratory distress seen at the CME CHU PZAGA Mahajanga. Rev Malg Ped 2018;1:34-41.
16. Li Y, Wang W, Zhang D. Maternal diabetes mellitus and risk of neonatal respiratory distress syndrome: A meta-analysis. Acta Diabetol. 2019;56:729-40.
17. Khan B, Allah Yar R, Khakwani AK, Karim S, Arslan Ali H. Preeclampsia Incidence and Its Maternal and Neonatal Outcomes With Associated Risk Factors. Cureus. 2022;14(11):e31143.
18. Mou AD, Barman Z, Hasan M, Miah R, Hafsa JM, Das Trisha A. Prevalence of preeclampsia and the associated risk factors among pregnant women in Bangladesh. Sci Rep. 2021;11(1):21339.
19. Kadhim Jasim S, Al-Momen H, Al-Asadi F. Maternal Anemia Prevalence and Subsequent Neonatal Complications in Iraq. Open Access Maced J Med Sci. 2020;8:71-5.
20. Menon KC, Ferguson EL, Thomson CD, Gray AR, Zodpey S, Saraf A, et al. Effects of anemia at different stages of gestation on infant outcomes. Nutrition. 2016; 32(1):61-5.
Download attachments: 10.4328.ACAM.21998
Latifa Mochhoury, Fatima Dahmanı, Touria Essayagh, Ikram Marc, Amina Barkat. Risk factors for neonatal respiratory distress : Moroccan retrospective study. Ann Clin Anal Med 2024;15(3):154-159
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
Effects of laparoscopic sleeve gastrectomy on early complications and 1-year comorbidity in a single center, single surgeon cohort of 342 patients
Medeni Sermet
Department of General Surgery, Faculty of Medicine, Medeniyet University, Goztepe Prof. Dr. Suleyman Yalcin City Hospital, Istanbul, Turkey
DOI: 10.4328/ACAM.22023 Received: 2023-10-25 Accepted: 2023-11-27 Published Online: 2024-01-05 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):160-164
Corresponding Author: Medeni Sermet, Department of General Surgery, Faculty of Medicine, Medeniyet University, Goztepe Prof. Dr. Suleyman Yalcin City Hospital, Dr. Erkin Street, Kadikoy, Istanbul, Turkey. E-mail: sermetmedeni@gmail.com P: +90 507 236 70 20 Corresponding Author ORCID ID: https://orcid.org/0000-0001-6119-0037
This study was approved by the Ethics Committee of İstanbul Medeniyet University (Date: 2022-07-22, No: 259)
Aim: The purpose of this study was to examine the 30-day morbidity and mortality rates after Laparoscopic Sleeve Gastrectomy (LSG) over a 5-year period.
Material and Methods: A retrospective analysis was conducted using prospectively collected data from patients who underwent LSG by the same surgeon between July 2017 and August 2022. The study identified LSG-related 30-day morbidity and mortality rates, as well as risk factors for 30-day morbidity. Furthermore, the impact of comorbidities on patients was evaluated at the 1-year follow-up.
Results: The study analysed the outcomes of 342 patients who underwent laparoscopic sleeve gastrectomy (LSG) over a period of five years. Early postoperative complications were experienced by 11.40% of patients (39 individuals), with six patients requiring blood transfusions, and 2.34% of patients (eight individuals) requiring readmission to hospital. The reoperation rate was 0.87% (three individuals) and the mortality rate was 0.29% (one individual). The study identified body mass index, diabetes, and hypertension as significant factors contributing to early postoperative complications. Technical term abbreviations were defined upon first use. At the six-month follow-up, patients demonstrated an average body weight loss of 62.9 ± 16.17%.
Discussion: The previously reported short-term safety of LSG, in terms of low 30-day postoperative morbidity and mortality rates, was confirmed by this study. Preoperative BMI, diabetes, and hypertension were found to be risk factors for 30-day morbidity and mortality. Additionally, an average 80% improvement in comorbidities was observed at one year.
Keywords: Sleeve Gastrectomy, Early Complications, Single Surgery, Comorbidity
Introduction
Obesity has been recognized as a widespread problem that is considered a public health crisis, due to the significant negative impact it has on one’s health [1]. Obesity surgery has proven to be a successful solution for patients who have not been able to achieve weight loss through non-surgical methods, and has shown to improve comorbidities related to obesity [2, 3]. This surgical intervention can be considered an important tool in the fight against obesity, and may be a vital treatment option for patients.
Laparoscopic sleeve gastrectomy (LSG) is a bariatric procedure that is rapidly increasing in popularity owing to its simple technical structure, safety and efficacy [4]. Based on a report by IFSO (International Society for the Surgery of Obesity) in 2018, this surgical procedure has been the most widely performed bariatric surgery since 2014 [5]. This can be considered a significant development in the treatment of obesity, and has become the preferred option for many patients for weight loss and health improvement.
Like any surgical intervention, LSG can be complicated by adverse events and involves risks that can result in death. The 30-day morbidity and mortality rates are important measures of the safety of surgical procedures and have been used for many years [6,7].
The aim of this study was to identify 30-day morbidity and mortality risk factors for LSG and to contribute to the development of strategies for perioperative management of patients undergoing this procedure. Focusing especially on at-risk groups may contribute to a safer implementation of LSG and help reduce adverse outcomes. This allows for the development of more effective and safe practices in the field of bariatric surgery.
Material and Methods
This study is based on data collected retrospectively from a consecutive group of patients who underwent LSG performed by a single surgeon over a 5-year duration. This study was approved by the Ethics Committee of Medeniyet University (Date: 22.07.2022 – No: 259).
The inclusion criteria were as follows: Patients aged 18-65 years, BMI = 35 – 39.90 kg/m2 with at least one obesity-related chronic comorbidity, and BMI > 40 kg/m2.
Exclusion criteria: Patients admitted for revisional bariatric surgery and those who opted for other bariatric surgical procedures, concurrent cholecystectomy, or herniation surgery were excluded. Furthermore, patients without follow-up data in the hospital registry system were excluded from the study.
Patients underwent a routine preoperative workup, including personal history taking, multidisciplinary clinical evaluation, laboratory investigations, and upper gastrointestinal endoscopy. A total of 72 patients were excluded based on the exclusion criteria.
Surgical Procedure
All patients underwent surgery according to the ERABS protocol [8]. After the usual preoperative preparations, the surgery was carried out under general anaesthesia. Pneumoperitoneum was induced, and a sleeve was inserted through a 38 Fr spark plug with resection taking place about 3-4 cm proximal to the pylorus. Subsequently, customary postoperative care was administered, and patients were advised to mobilise early and given a postoperative dietary regime, supplement plan, and schedule of follow-up visits. Patients were also informed to seek medical attention in the event of any adverse reaction.
Demographic characteristics, operative details, and perioperative events were recorded and analyzed.
Thirty-day postoperative data was available for all patients while excluding one case of death, resulting in 341/342 patients (99.7%) having available 6-month follow-up data. Moreover, data for 338 patients (98.8%) were accessible at the 1-year follow-up..
Statistical Analysis
Patient data were analyzed using the Statistical Package for Social Sciences (SPSS) software (IBM Corp., Armonk, NY, USA), version 28. Numerical data were presented using mean, standard deviation, and range, while frequencies and percentages were used for categorical values. To identify early postoperative morbidity risk factors, a binary logistic regression analysis was conducted, with statistical significance set at 0.05.
Results
This investigation comprised 342 individuals who received LSG carried out by a single surgeon from July 2017 to August 2022. The average age of the patients was 37.89 ± 11.36 years, with a higher prevalence among women (82.74%, n = 283). The preoperative weight ranged from 96 to 192 kg, with a mean of 126.93 ± 23.32, and preoperative BMI ranged from 37.7 to 69.2 kg/m2. On average, the body mass index (BMI) of patients was 45.3 ± 8.06 kg/m2 and the excess body weight (EBW) before surgery ranged from 38.3 to 122.1 kg with a mean of 62.1 ± 4.64 kg. The patients had accompanying diseases, such as dyslipidemia, hypertension, type 2 diabetes, and obstructive sleep apnea (Table 1).
The preoperative prophylaxis against deep vein thrombosis (DVT) was given subcutaneously as Enoxaparin (0.4 mL). Two weeks of anticoagulant therapy for DVT prophylaxis was continued. Table 2 shows the distribution of complications.
The total surgical duration ranged from 34 to 96 minutes, with an average of 52.18 ± 25.22. Early postoperative side effects were observed in 39 (11.42 %) patients. Complications occurred in 25 patients before discharge and in 14 within the first 30 days after discharge. The total hospital stay varied from 2 to 16 days.
Ten patients had intra-abdominal hemorrhage in the postoperative period, which developed within the first 24 h. Four patients were treated conservatively, while six patients with hemodynamic instability were treated with erythrocyte suspension.
Hematoma was present at the trocar site in three patients, two at the 10 mm port site, and one at the 15 mm port site, and the patients were treated with local drainage by suturing.
Postoperative leakage developed in three patients, with one occuring within the first 24 hours and the other two within 48 hours. These three patients typically developed leakage at the His angle and were treated primarily through open laparotomy. Two patients experienced an intraoperative leakage. In both patients, the nasogastric catheter was found in the stapler line, and intraoperative primary repair was performed. Intraoperative hemorrhage was observed in four patients, which was due to gastrolienal ligament dissection in two patients, splenic capsule rupture in one patient, and retractor entry due to left liver injury. Three patients had intra-abdominal abscesses complicated with hematoma formation and were treated with percutaneous abscess drainage under the umbrella of antibiotic therapy. Three patients had hematoma at the trocar site, one patient developed hematoma after coughing at the 10 mm port site, and the other two hematomas developed after intraoperative inadequate drainage at the 15 mm port site. All three patients were treated with local hematoma drainage by suturing.
A 36-year-old woman with a BMI of 53.1 kg/m2, type 2 diabetes (T2D), hypertension and dyslipidemia developed portal vein thrombosis on day 20. She underwent thrombolysis under radiological monitoring, followed by continuous thromboprophylaxis. A 52-year-old male patient with a BMI of 62.5 kg/m2, T2D, and hypertension developed bleeding in the first 24 hours postoperatively and was administered a total of 3 units of erythrocyte suspension. However, pulmonary embolism developed on the 14th postoperative day despite thromboprophylaxis in the patient who had inadequate mobilization afterwards, and the patient was transferred to the intensive care unit.
During one year of regular ultrasonography follow-up, herniation developed at the trocar entry site in 3 cases. Two of the herniations were at the 15 mm port site, and one was at the 10 mm port site. Richter’s hernia developed at the 10 mm port site on the left upper part of the umbilicus on the 8th day and small bowel resection and anastomosis was performed at 120 cm because the jejunum anus was necrotic. Another patient had volvulus in the incisura angularis due to persistent vomiting on postoperative day 20, which was resolved with gastric bypass.
Factors contributing to early postoperative adverse events are shown in Table 3.
Binary logistic regression analysis showed that BMI (OR = 1.078, CI 0.962 – 1.2, p = 0.048), T2D (OR = 0.88, CI 0.762 – 0.952, p = 0.037), hypertension (OR = 0.091, CI 0.021 – 0.469, p = 0.003), and reoperation (OR = 0.55, CI 0.137 – 0.982, p = 0.058) were significant predictors of early postoperative morbidity.
At 6 months follow-up, the mean BMI was 35.82 ± 5.69 kg/m2. The mean EBWL% was 62.9 ± 16.17%. The mean BMI was 28.92 ± 5.32 kg/m2, and the mean EBWL% was 81.57 ± 16.15% at 1 year post-op. Considering associated comorbidities, 96.05% of patients with dyslipidemia (n=84), 72.31% of patients with hypertension (n=89), 89.12% of patients with diabetes (n=82), and 100% of patients with obstructive sleep apnea (n=19) had complete recovery.
Discussion
This retrospective cohort study was conducted to evaluate the safety and efficacy of bariatric surgery, particularly LSG. These findings suggest that bariatric surgery is an important treatment option for obese patients. In this retrospective cohort study of 342 patients who underwent LSG by a single surgeon in our center, the early postoperative morbidity, reoperation, local intervention, rehospitalization, and mortality rates were 11.40%, 0.87%, 0.87%, 0.87%, 2.34%, and 0.29%, respectively.
Despite the reported safety of bariatric surgery, varying rates of perioperative complications have also been reported. Patients undergoing bariatric surgery are at high risk of developing early postoperative complications. Therefore, it is important to identify the risk factors for early postoperative complications to optimize pre- and post-operative care. In the literature, 30-day complication rates after bariatric surgery range from 0.2% to 5%, and reoperation rates range from 0.6% to 1.1% [9, 10]. Mortality rates were found to be between 0 and 0.3% [11]. Although our total complication rate in the first 30 days seems to be high, this may be because we included data such as local drainage, intraoperative hemorrhage, and dehydration, which do not prolong the patient’s hospitalization period. Furthermore, mortality and reoperation are consistent with the literature. These data were collected in order to expand the scope of our study and provide a more thorough evaluation. Additionally, including this data allows for a more comprehensive assessment of the success of surgical interventions, rather than solely focusing on severe complications. Timely diagnosis and management of adverse events that may occur during the 30-day postoperative follow-up directly affect the success of surgery. The mortality rate was 0.13% in a database of 3.6 million cases [12]. In the present study, the single-case mortality rate was 0.29%. This finding is consistent with the general literature. These results emphasize the relative safety of bariatric surgery and reflect the advances in minimizing the risks associated with this procedure.
In our study, the patient’s BMI, presence of hypertension, and diabetes were found to be predictors of early perioperative morbidity. A greater risk of disease has been documented in patients with a BMI of 40.0 or higher, particularly when they have both hypertension and diabetes. The risk of disease in this group is 15% [12, 13]. These results suggest that a patient’s preoperative status and metabolic syndrome have a significant impact on the risk of complications after bariatric surgery. Furthermore, it has been observed that patients with a higher BMI have an increased rate of leakage, and hypertension is associated with an increased risk of early bleeding [13].
In addition to these risk factors, the learning curve for specialized surgery also affects early complications. In a survey conducted in Italy, 54.3% of bariatric surgeons stated that they had never received bariatric surgery training during their specialty training [14]. When we assigned our study patients to three consecutive equal groups, 19 (48.7%) complications were observed in the first 100 cases, and serious complications, such as fistula and bleeding, were observed in this period. On the other hand, forgetting to withdraw the orogastric catheter, which seems to be a preventable complication, was also observed in the first 100 cases. It is possible that many surgeons have learned the principles of surgery through courses and short training sessions. This finding suggests that complications may occur more frequently during learning.
The rates of comorbidities in the first 6 months and the first year after LSG in our patients were comparable to those previously reported [15,16]. The rates of improvement in dyslipidemia, hypertension, and T2DM at 6 months and 1 year were > 80%. In a previous study, dyslipidemia and hypertriglyceridemia improved by 45% and 86%, respectively [17].
Unlike the literature, we found that all of our patients with apnea syndrome (all 19 cases) had resolution of their complaints and were device-free [18, 19]. We believe that this high rate is due to the low number of patients with apnea syndrome in our cohort and the fact that we evaluated the patients’ drug and device use rather than postoperative measured data.
The postoperative bleeding rate after LSG has been reported to vary between 2-4% [20, 21]. The rate in our study was 2.9%. Although these rates are similar to those reported in the literature, it is worth noting that we still found them high. We did not apply any reinforcement to the stapler line in ⅔ of the cases. Later, with increasing experience, we observationally decreased the bleeding rates by selecting patients with bleeding risk, using postoperative tranexamic acid, and, in some cases, using reinforcing sutures on the stapler line. However, we did not transfuse blood products in patients who developed bleeding unless vital signs significantly deteriorated. Adequate fluid resuscitation and blood product supplementation when necessary for the diagnosis of bleeding can solve these problems. Reoperation should be avoided, whenever possible because possible complications of the new operation may cause more difficult and longer treatment processes [22].
Leakage is an important and frightening complication. Most leaks occur in the first week and reported rates are between 0.5-5% [23, 24].
Our results were consistent with those reported in the literature. Based on our experience, the leak rate decreases with increasing experience and follow-up. All our leakage cases were within the first 20 consecutive cases. Although endoscopic stenting for leaks is prioritized in the literature, we performed early laparotomy and primary repair because of individual concerns. In the following cases, applying clips to the stapler joints, paying particular attention to the His angle, and applying clips to the last point in the fundus region were found to be very effective in preventing leaks.
During one year of regular ultrasonography follow-up, three patients developed herniation at the trocar entry site. Two of these herniations occurred at the 15 mm port entry, and the other at the 10 mm port entry. Trocar site herniation occurs in 0.1-2% [24]. Richter’s hernia developed on the 8th day especially at the 10 mm port entry in the upper left region of the umbilicus and small bowel resection and anastomosis were required because the jejunum anus was necrotic. Another patient was diagnosed with volvulus in the incisura angularis on postoperative day 20 because of persistent vomiting and underwent gastric bypass surgery. During the surgical operation, regardless of surgical experience, in two patients, the orogastric catheter was forgotten before stapling, and the catheter remained in the stapler line. Fortunately, this error was quickly recognized, and a primary repair was performed. This highlights the importance of synchronization within the surgical team. It should also be kept in mind that due to staff turnover, every step should be checked, and the staff should be careful, even though it may seem like a continuous routine.
The most common reason for readmission in our patient group was dehydration [25]. This occurs because of patients’ fear after LSG and difficulty in adapting to their new lifestyle. In this case, patients should be hospitalized before their vital signs worsen, appropriate nutrition should be provided, and vitamin and electrolyte disorders should be corrected.
Based on the results of our study, we confirmed that LSG is an effective method for weight loss and the improvement of obesity-related comorbidities. The first 30-day complication rates are consistent with those in the current literature, and the success rate has increased in proportion to timely complication management and increasing experience.
Limitations: The study was limited to a single center with a single surgeon and a relatively small number of cases. The advantages are that it included all cases in the learning curve and experience gaining phase and that the records and treatments were recorded in detail from a single source.
In conclusion, this study shows that bariatric surgery is a safe and effective treatment option for obese patients. Preoperative review can be an important tool for surgeons to identify risk factors. Although undesirable, encountering and managing a range of complications within limits have contributed to lower complication rates by making subsequent surgeries more careful and safer. However, while more research is needed, keeping records without omitting minor details will help us better understand the long-term effectiveness of bariatric surgery and the risk of complications.
In summary, this study affirms that the Laparoscopic Sleeve Gastrectomy (LSG) procedure is linked with minimal complications during the 30-day postoperative period. These findings highlight the potential for LSG as a dependable and advantageous alternative for addressing obesity in clinical settings. Specifically, the low incidence of complications may assist patients in recovering postoperatively and provide health practitioners with a secure surgical choice. Additionally, the results indicate that patients with other obesity-related health issues, such as type 2 diabetes or hypertension, have a reduced risk of postoperative complications. This information could serve as a significant source of guidance for patients contemplating this procedure. The findings of this research can enhance the approach to managing obesity and its related comorbidities, thus enabling healthcare professionals to make more informed choices.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: None
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Stenberg E, Dos Reis Falcão LF, O’Kane M, Liem R, Pournaras DJ, Salminen P, et al. Guidelines for Perioperative Care in Bariatric Surgery: Enhanced Recovery After Surgery (ERAS) Society Recommendations: A 2021 Update. World J Surg. 2022;46(4):729-51.
2. Eisenberg D, Shikora SA, Aarts E, Aminian A, Angrisani L, Cohen RV, et al. 2022 American Society of Metabolic and Bariatric Surgery (ASMBS) and International Federation for the Surgery of Obesity and Metabolic Disorders (IFSO) Indications for Metabolic and Bariatric Surgery. Obes Surg. 2023;33(1):3-14.
3. Buchwald H. The evolution of metabolic/bariatric surgery. Obes Surg. 2014;24(8):1126-35.
4. Chang SH, Freeman NLB, Lee JA, Stoll CRT, Calhoun AJ, Eagon JC, et al. Early major complications after bariatric surgery in the USA, 2003-2014: a systematic review and meta-analysis. Obes Rev. 2018;19(4):529-37.
5. Stone G, Samaan JS, Samakar K. Racial disparities in complications and mortality after bariatric surgery: A systematic review. Am J Surg. 2022;223(5):863-78.
6. El Masry MAMA, Rahman IA. Perioperative Morbidity and Mortality of Laparoscopic Sleeve Gastrectomy (LSG) in a Single-Surgeon Experience on 892 Patients Over 11 Years. World J Surg. 2023;47(11):2809-15.
7. Poole M, Fasola L, Zevin B. Management of Complications After Bariatric Surgery: As Survey of Comfort and Educational Needs of General Surgeons in Ontario, Canada. Obes Surg. 2022;32(7):2407-16.
8. Robertson AGN, Wiggins T, Robertson FP, Huppler L, Doleman B, Harrison EM, et al. Perioperative mortality in bariatric surgery: meta-analysis. Br J Surg. 2021;108(8):892-7.
9. Kim J, Azagury D, Eisenberg D, DeMaria E, Campos GM. American Society for Metabolic and Bariatric Surgery Clinical Issues Committee. ASMBS position statement on prevention, detection, and treatment of gastrointestinal leak after gastric bypass and sleeve gastrectomy, including the roles of imaging, surgical exploration, and nonoperative management. Surg Obes Relat Dis. 2015;11(4):739-48.
10. Noria SF, Shelby RD, Atkins KD, Nguyen NT, Gadde KM. Weight Regain After Bariatric Surgery: Scope of the Problem, Causes, Prevention, and Treatment. Curr Diab Rep. 2023;23(3):31-42.
11. Grieco A, Huffman KM, Cohen ME, Hall BL, Morton JM, Ko CY. The Metabolic and Bariatric Surgery Accreditation and Quality Improvement Program bariatric surgical risk/benefit calculator: 30-day risk. Surg Obes Relat Dis. 2021;17(6):1117-24.
12. Mahendran V, Ricart P, Levine F, White E, Abolghasemi-Malekabadi K, Williams M, et al. Bariatric Surgery as a Viable Treatment for Idiopathic Intracranial Hypertension: a Case Series and Review of Literature. Obes Surg. 2021;31(10):4386-91.
13. Mollan SP, Mitchell JL, Ottridge RS, Aguiar M, Yiangou A, Alimajstorovic Z, et al. Effectiveness of Bariatric Surgery vs Community Weight Management Intervention for the Treatment of Idiopathic Intracranial Hypertension: A Randomized Clinical Trial. JAMA Neurol. 2021;78(6):678-86.
14. Wyszomirski K, Walędziak M, Różańska-Walędziak A. Obesity, bariatric surgery and obstructive sleep Apnea: A narrative literature review. Medicina (Kaunas). 2023;59(7):1266-71.
15. Martin-Rodriguez E, Guillen-Grima F, Martí A, Brugos-Larumbe A. Comorbidity associated with obesity in a large population: The APNA study. Obes Res Clin Pract. 2015;9(5):435-47.
16. Spiro C, Bennet S, Bhatia K. Meta-analysis of patient risk factors associated with post-bariatric surgery leak. Obes Sci Pract. 2022;9(2):112-26.
17. Kumar SB, Hamilton BC, Wood SG, Rogers SJ, Carter JT, Lin MY. Is laparoscopic sleeve gastrectomy safer than laparoscopic gastric bypass? A comparison of 30-day complications using the MBSAQIP data registry. Surg Obes Relat Dis. 2018;14(3):264-269.
18. Syn NL, Cummings DE, Wang LZ, Lin DJ, Zhao JJ, Loh M, Koh ZJ, Chew CA, Loo YE, Tai BC, Kim G, So JB, Kaplan LM, Dixon JB, Shabbir A. Association of metabolic-bariatric surgery with long-term survival in adults with and without diabetes: A one-stage meta-analysis of matched cohort and prospective controlled studies with 174 772 participants. Lancet. 2021;397(10287):1830-41.
19. Kristensson FM, Andersson-Assarsson JC, Svensson P-A, Carlsson B, Peltonen M, Carlsson LMS. Effects of bariatric surgery in early- and adult-onset obesity in the prospective controlled Swedish Obese Subjects study. Diabetes Care. 2020;43(4):860–6.
20. Argyriou K, Parra-Blanco A. Reconstructing the puzzle of the role of therapeutic endoscopy in the management of post-bariatric surgery complications. World J Gastroenterol. 2022;28(23):2633-5.
21. Mierzwa AS, Mocanu V, Marcil G, Dang J, Switzer NJ, Birch DW et al. Characterizing Timing of Postoperative Complications Following Elective Roux-en-Y gastric Bypass and Sleeve Gastrectomy. Obes Surg. 2021;31(10):4492-501.
22. Yanari S, Sasaki A, Umemura A, Ishigaki Y, Nikai H, Nishijima T, et al. Therapeutic effect of laparoscopic sleeve gastrectomy on obstructive sleep apnea and relationship of type 2 diabetes in Japanese patients with severe obesity. J Diabetes Investig. 2022;13(6):1073-85.
23. Ohta M, Kasama K, Sasaki A, Naitoh T, Seki Y, Inamine S, et al. Current status of laparoscopic bariatric/metabolic surgery in Japan: The sixth nationwide survey by the Japan Consortium of Obesity and Metabolic Surgery. Asian J Endosc Surg. 2021;14(2):170-77.
24. Ahlqvist S, Edling A, Alm M, Dackhammar JB, Nordin P, Cengiz Y. Trocar site hernia after gastric sleeve. Surg Endosc. 2022;36(6):4386-91
25. Habas E, Farfar K, Errayes N, Rayani A, Elzouki AN. Wernicke Encephalopathy: An Updated Narrative Review. Saudi J Med Med Sci. 2023;11(3):193-200.
Download attachments: 10.4328.ACAM.22023
Medeni Sermet. Effects of laparoscopic sleeve gastrectomy on early complications and 1-year comorbidity in a single center, single surgeon cohort of 342 patients. Ann Clin Anal Med 2024;15(3):160-164
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
Comparison of the cyclic fatigue resistance of the original reciproc files with equipollents
İhsan Furkan Ertuğrul, Samet Tosun
Department of Endodontics, Faculty of Dentistry, Pamukkale University, Denizli, Turkey
DOI: 10.4328/ACAM.22025 Received: 2023-10-26 Accepted: 2023-11-27 Published Online: 2024-01-09 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):165-168
Corresponding Author: İhsan Furkan Ertuğrul, Department of Endodontics, Faculty of Dentistry, Pamukkale University, Denizli, Turkey. E-mail: furkanertugrul@gmail.com P: +90 536 517 46 77 Corresponding Author ORCID ID: https://orcid.org/0000-0001-7583-6679
This study was approved by the Ethics Committee of Pamukkale University (Date: 2023-08-17, No:E-60116787-020-406510)
Aim: In this study, cyclic fatigue resistance of Reciproc R25, Golden Star RC, NIC RC and RO files were compared. The present study aimed to compare the cyclic fatigue strength of the original RC files and their equivalent GS, NIC and RO files.
Material and Methods: In this study, 100 files, 25 in each group, were subjected to cyclic fatigue test on the artificial canal on stainless steel block with 60° curvature, 5 mm radius and 1.5 mm depth. The files in all groups were rotated until they were broken in the artificial canal using the Reciproc ALL program. The time to break was recorded with a 1/100 second precision stopwatch for each file and the total number of rounds was calculated. Number of rounds of files and the lengths of fractures were recorded then evaluated statistically (P<.05). The files were examined by scanning electron microscopy (SEM) images to evaluate the fracture type.
Results: As a result of the data obtained from the study, the RC file was statistically the most resistant to cyclic fatigue (p <.05). GS and NIC files were found to be more resistant than RO (p <.05).
Discussion: Based on the data derived from this study, it is evident that the RC file exhibited the highest cyclic fatigue resistance, as indicated by statistically significant results. In comparison, GS and NIC files displayed greater resistance to cyclic fatigue than RO files. The examination of SEM images further corroborated the presence of cyclic fatigue fractures in the tested files. Notably, GS, NIC, and RO files, despite sharing a similar shape with the RC, failed to match the original RC file’s level of resistance against cyclic fatigue, underscoring the importance of considering file composition and heat treatment in enhancing resistance.
Keywords: Cyclic Fatigue, Golden Star, NIC, Reciproc, Recip-One
Introduction
Today, nickel-titanium rotary instruments (NiTi) are often used for mechanical instrumentation of root canals. NiTi files have gained popularity due to their lower incidence of complications, such as zipping, stepping, and perforation, compared to traditional stainless steel files [1, 2]. Despite the advantages of NiTi files, the risk of fracture should be considered by clinicians, especially in curved root canals [3]. A broken NiTi instrument in the root canal system adversely affects the success of root canal treatment (RCT). The main reasons for breaking NiTi files are cyclic fatigue and torsional fatigue [4, 5]. In cyclic fatigue, a specific area of NiTi files is subjected to compression and tensile forces in each cycle and eventually breaks [6]. Recently, several methods have been developed to increase the cyclic fatigue resistance of NiTi rotary files, such as various heat treatments, different cross-sectional designs and application of surface electro polishing processes [7]. According to a previously presented study [8], it has been reported that root canals can be enlarged by a single file with reciprocal motion. The kinematics of the file in a reciprocal motion are clockwise and counterclockwise. It has been found that reciprocal motion causes the resistance of the files to increase [9, 10]. The Reciproc (VDW motion to shape root canals in the clinic. Many studies have reported the superior ability of this system to root canal treatment [11] and retreatment [12, 13]. The original Reciproc (RC; VDW, Munich, Germany) file has M-Wire technology and is reported to have a very high cyclic resistance with reciprocal motion [14-16]. The Reciproc file is reported to be in austenitic phase during clinical use [17]. The original Reciproc R25, which is planned to be used in the study, and its equipollent productions are Golden Star RC 25 (GS; Golden Star LTD, Shenzhen, China), NIC RC 25 (NIC; Superline Thecnology, Shenzhen, China), Recip-One (RO; Rogin Medical LTD, Shenzhen, China) root canal files. In the literature, there is no study that compares the RC file’s cyclic fatigue resistance with its equipollents (GS, NIC and RO). The purpose of the study is to compare the cyclic fracture resistance between RC files and its equipollents GS, NIC and RO files. The null hypothesis of the study is that the cyclic fatigue resistance of the original RC file resides in the same cyclic resistance as equipollent GS, NIC and RO files.
Material and Methods
Stainless steel artificial root canal with a 5 mm radius, 1.5 mm depth, and a 60° curvature were used described in the present study as a previous study [6]. In accordance with the limitations of the study, each type of NiTi file was used as a group. In the study, a total of 4 groups of 100 files was examined with a stereo microscope (SMZ1500, Nikon Corp., Tokyo, Japan), and damaged and/or deformity files were replaced with new ones. NiTi file types were tested. RC R25 and its equipollent production were selected as GS 25, NIC 25 and RO 25 (n = 25). The root canal files were tested in the “Reciproc All” program with the 6:1 reductive VDW Silver Reciproc endodontic motor (VDW). In order to reduce friction during the experiments, lubricant oil (WD-40, Milton Keynes, England) was used in the artificial root canal. The rotational time of the file in the artificial root canal was measured by stopwatch until it was broken (Fig. 1). Then, the rotational times until break were calculated in seconds and multiplied by the number of turns per second, and the number of cyclic failure (NCF) was found. The number of rounds per minute was calculated by the manufacturer to claim 300 rpm for the “Reciproc All” program. As a result of cyclic fatigue, the fractures of the files were measured by means of calipers to determine whether the areas affected by the cyclic force were the closest to each other. After the cyclic fatigue test, 8 pieces of fractured files (2 pieces from each group) were observed using a scanning electron microscope (SEM) (Zeiss Supra 40VP, Carl Zeiss SMT Inc., Oberkochen, Germany) to determine the fracture types under 500X and 3000X magnification.
Statistical Analysis
All statistical analyses were performed using SPSS 24.0 (SPSS Inc., Chicago, IL, USA). Shapiro–Wilk tests were used for testing normality. Group comparisons were conducted through the utilization of analysis of variance (ANOVA) followed by post-hoc Tukey tests. P-value < .05 was considered statistically significant. This study was approved by the Ethics Committee of Pamukkale University (Date: 2023-08-17, No:E-60116787-020-406510).
Results
The cyclic fatigue resistance of the original RC file was found to be significantly higher than the equipollents GS, NIC and RO files (P<.05). While there was no statistically difference between NIC and GS files in terms of cyclic fatigue resistance, the resistance of RO file was statistically significantly lower than NIC and GS files (P<.05, Table 1). There was no significant difference in the mean length of the fractured parts (Table 1), so that the fragments exposed to compression and tensile forces were identical. SEM analysis of the fractured surfaces of the files have typical cyclic fatigue crack origins (Figure 2).
Discussion
In the present study, the cyclic fatigue resistance simulations of the RC file were compared with GS, NIC and RO files. Files were tested in a 5 mm diameter artificial canal with a 60° curve angle. In this study, when the lengths of the broken parts were examined, no statistically significant difference was found between the lengths of the parts. This finding shows that the files tested are in a standard position in the artificial canal and that the area from which the force comes corresponds to the same region [18]. It has been reported that the reciprocal motion increases the resistance of the files against cyclic fatigue considerably [9, 14]. In this study, the files using with reciprocal motion were compared with each other. According to statistical data, the cyclic resistance of the original RC file was found to be significantly better than its equipollents GS, NIC and RO files. As a result of the presented study, the null hypothesis was rejected. The use of NiTi files with endodontic motors in the shaping of the root canals shortened the treatment process and also reduced the complications during the treatment (perforations, change working length, apical transportation) [19].
During RCT, root canal files broken during instrumentation of the root canals adversely affect the success of the treatment. The main causes broken of root canal files are torsional and cyclic fatigue [19]. The cyclic fatigue resistance of the files , which have been used in the present study, was compared. Root canal files generally work with 2 kinds of motion. The first is a continuous rotation motion, which has a continuous clockwise rotation movement while the second is the reciprocating motion, which is clockwise and counterclockwise at various angles. In previous studies [9, 14], it has been observed that the resistance of the files at curved canals with reciprocal motion increases significantly. It is also reported that RC files have higher resistance than classic files such as Mtwo and ProTaper although they have similar cross section [20]. In the literature, there is no study that compares RC files resistance with its own equipollents. While GS and NIC files in the semi-martensitic phase with heat treatment, the RO file is made of super-elastic NiTi alloy which has not been heat treated. Heat treated files such as GS and NIC, which are close to the gold color, have been very popular lately, and many studies have been done about these type files [21]. Almost all of these studies, which are also called gold alloy, have been reported to increase the resistance of the files to the fracture as a result of the heat treatment they are subjected to after shaping the files [22-24]. In the present study, it was found that the heat treated GS and NIC files, which are called gold alloys, were found to have statistically less cyclic fatigue resistance compared to the RC file which was not heat treated and only called M-Wire (P<.05). This result is not compatible with previous studies [22-24] for heat-treated files. This is because GS and NIC files are produced as equipollents of RC files, and they do not belong to a recognized company and that the quality of production is lower. However, when the equipollent production files are compared among themselves, the cyclic fatigue resistance of the RO file, which is only super elastic, is statistically lower than the heat treated GS and NIC files [22-24]. The results of the tests showed that the lengths of the broken file parts were very close to each other, indicating that the regions where the cyclic fatigue intensity was the same, so that the metal volumes affected were close to each other.
Conclusion
In the present study, the RC file have statistically the highest cyclic fatigue resistance, and the RO file has the lowest. However, performances of GS and NIC files cannot be considered bad. Notably, while GS, NIC, and RO files share a similar shape with the RC file, they fall short of replicating its original level of resistance against cyclic fatigue. This emphasizes the significance of considering file composition and heat treatment in enhancing cyclic fatigue resistance.
Acknowledgment
This project, labeled 2019HZDP003, was supported by Pamukkale University Scientific Research Projects Unit . I would like to thank Hande ŞENOL for her contributions to the statistical analysis of the data.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: This study was supported by Pamukkale University-BAP-HZD program (2019HZDP003).
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Langaliya AK, Kothari, AK, Surti, NR, Patel, AR, Doshi, PR, Pandya DJ. In vitro comparative evaluation of dentinal microcracks formation during root canal preparation by different nickel-titanium file systems. Saudi Endod. 2018;8(3):183-8.
2. Cheung GS. Instrument fracture: mechanisms, removal of fragments, and clinical outcomes. Endod Topics. 2007;16(1):1-26.
3. Orhan, EO, Ertuğrul İF, Tosun, S. Monotonic torsional resistance and fatigue resistance of novel SCOPE RS instruments. Microsc ResTech. 2021;84(1):62-70.
4. Pedullà E, Lo Savio F, Boninelli S, Plotino G, Grande NM, La Rosa G, et al. Torsional and Cyclic Fatigue Resistance of a New Nickel-Titanium Instrument Manufactured by Electrical Discharge Machining. J Endod. 2016;42(1):156-9.
5. Gambarini G, Cicconetti A, Di Nardo D, Miccoli G, Zanza A, Testarelli L, et al. Influence of different heat treatments on torsional and cyclic fatigue resistance of nickel–titanium rotary files: A comparative study. Applied Sci. 2020;10(16):5604.
6. Gambarini G, Grande NM, Plotino G, Somma F, Garala M, De Luca M, et al. Fatigue resistance of engine-driven rotary nickel-titanium instruments produced by new manufacturing methods. J Endod. 2008;34(8):1003-5.
7. Peters OA, Gluskin A, Weiss R, Han J. An in vitro assessment of the physical properties of novel Hyflex nickel–titanium rotary instruments. Int Endod J. 2012;45(11):1027-34.
8. Yared G. Canal preparation using only one Ni‐Ti rotary instrument: preliminary observations. Int Endod J. 2008;41(4):339-44.
9. Plotino G, Ahmed HMA, Grande NM, Cohen S, Bukiet F. Current assessment of reciprocation in endodontic preparation: A comprehensive review—part II: properties and effectiveness. J Endod. 2015;41(12):1939-50.
10. De-Deus G, Leal Vieira VT, Nogueira da Silva EJ, Lopes H, Elias CN, Moreira EJ. Bending resistance and dynamic and static cyclic fatigue life of Reciproc and WaveOne large instruments. J Endod. 2014;40(4):575-9.
11. Plotino G, Rubini AG, Grande NM, Testarelli, Gambarini G. Cutting Efficiency of Reciproc and WaveOne Reciprocating Instruments. J Endod. 2014;40(8):1228-30.
12. Ozyurek T, Yilmaz K, Uslu G. Shaping Ability of Reciproc, WaveOne GOLD, and HyFlex EDM Single-file Systems in Simulated S-shaped Canals. J Endod. 2017;43(5):805-9.
13. Zuolo AS, Mello JE, Cunha RS, Zuolo ML, Bueno CES. Efficacy of reciprocating and rotary techniques for removing filling material during root canal retreatment. Int Endod J. 2013;46(10):947-53.
14. Pedullà E, Grande NM, Plotino G, Gambarini G, Rapisarda E. Influence of continuous or reciprocating motion on cyclic fatigue resistance of 4 different nickel-titanium rotary instruments. J Endod. 2013;39(2):258-61.
15. Ahn S-Y, Kim H-C, Kim E. Kinematic effects of nickel-titanium instruments with reciprocating or continuous rotation motion: A systematic review of in vitro studies. J Endod. 2016;42(7):1009-17.
16. You SY, Bae KS, Baek SH, Kum KY, Shon WJ, Lee W. Lifespan of one nickel-titanium rotary file with reciprocating motion in curved root canals. J Endod. 2010;36(12):1991-4.
17. Plotino G, Grande NM, Testarelli L, Gambarini G, Castagnola R, Rossetti A et al. Cyclic Fatigue of Reciproc and Reciproc Blue Nickel-titanium Reciprocating Files at Different Environmental Temperatures. J Endod. 2018;44(10):1549-52.
18. Plotino G, Grande NM, Cordaro M, Testarelli L, Gambarini G. A review of cyclic fatigue testing of nickel-titanium rotary instruments. J Endod. 2009;35(11):1469-76.
19. Kakar S, Dhingra A, Sharma H. Shaping potential of manual NiTi K-File and rotary ProTaper and analyzing the final outcome of shaped canals using CT. J Contemp Dent Pract. 2013;14(3):451-5.
20. da Frota MF, Espir CG, Berbert FL, Marques AA, Sponchiado-Junior EC, Tanomaru-Filho M, et al. Comparison of cyclic fatigue and torsional resistance in reciprocating single-file systems and continuous rotary instrumentation systems. J Oral Sci. 2014;56(4):269-75.
21. Keskin C, Inan U, Demiral M, Keleş A. Cyclic fatigue resistance of Reciproc Blue, Reciproc, and WaveOne Gold reciprocating instruments. J Endod. 2017;43(8):1360-3.
22. Özyürek T. Cyclic fatigue resistance of Reciproc, WaveOne, and WaveOne Gold nickel-titanium instruments. J Endod. 2016;42(10):1536-9.
23. Goo HJ, Kwak SW, Ha JH, Pedullà E, Kim HC. Mechanical properties of various heat-treated nickel-titanium rotary instruments. J Endod. 2017;43(11):1872-7.
24. Elnaghy A, Elsaka S. Cyclic fatigue resistance of XP-endo Shaper compared with different nickel-titanium alloy instruments. Clin Oral Investig. 2018;22(2):1433-7.
Download attachments: 10.4328.ACAM.22025
İhsan Furkan Ertuğrul, Samet Tosun. Comparison of the cyclic fatigue resistance of the original reciproc files with equipollents. Ann Clin Anal Med 2024;15(3):165-168.
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
Immunomodulatory potential of silver nanoparticles and therapeutic effect against doxorubicin-induced-cardiotoxicity in rats
Nesrin I. Tarbiah 1, Fares K. Khalifa 1, 2, Nuha A. Alkhattabi 1, Reem F. Ghazali 3, Reem Y. Alzahri 4, Sahar A. Alkhodair 1, Reham A. Shindi 5, Ahd A. Mansour 6
1 Department of Biochemistry, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia, 2 Department of Biochemistry and Nutrition, Faculty of Women for Arts, Science and Education, Ain Shams University, Cairo, Egypt, 3 Department of Clinical Biochemistry, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia, 4 Department of Biology, Faculty of Science, University of Jeddah, Saudi Arabia, 5 Department of Clinical Medical Laboratories, Faculty of Applied Medical Science, Inaya Medical Colleges, Riyadh, Saudi Arabia, 6 Department of Medical Laboratory Sciences, Fakeeh College for Medical Sciences, Jeddah, Saudi Arabia
DOI: 10.4328/ACAM.22032 Received: 2023-11-03 Accepted: 2023-12-11 Published Online: 2024-01-12 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):169-175
Corresponding Author: Nesrin I. Tarbiah, Department of Biochemistry, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia. E-mail: ntarabah@kau.edu.sa P: +96 650 567 12 53 Corresponding Author ORCID ID: https://orcid.org/0000-0002-8706-2663
This study was approved by the Ethics Committee of Sciences Academy of Experimental Research (Date: 2023-05-15, No: 44121)
Aim: The present study was designed to determine the therapeutic effects and immunomodulatory potential of silver nanoparticles (AgNPs) at two doses on cardiotoxicity induced by doxorubicin (DOX).
Material and methods: Sixty rats were randomized into six equal groups as follows: G1; Control, G2; NP1, rats received AgNPs (4 mg/kg b.w), G3; NP2, rats received AgNPs (8 mg/kg b.w), G4; cardiotoxicity group (DOX); rats received intraperitoneal single injection of DOX at dose of (30 mg/kg; i.p.), G5; DOX+ NP1, G6; DOX+ NP2. AgNPs was administered intraperitoneally for 4 weeks.
Results: AgNPs showed immunomodulatory potential as shown by an increase in blood leukocytes, lymphocytes, and neutrophils counts and the decrease in serum proinflammatory cytokines, and oxidative stress markers. Cardio- therapeutic effects of AgNPs are shown by the decrease in biochemical markers of cardiac toxicity, namely, CPK, LDH, TropT, and ACL. DOX-treated rats showed significantly higher total cholesterol, triglycerides, total phospholipids, and LDL-c. Administration of AgNPs as therapeutic (4 and 8 mg/kg b.w) improved these parameters.
Discussion: Depending on the immunomodulatory activity of Ag NPs, therapeutic effect on the heart against cardiotoxicity induced by doxorubicin was shown by an increase in blood leukocytes count, decrease in proinflammatory cytokines, and oxidative stress biomarkers.
Keywords: Silver Nanoparticles, Cardiotoxicity, Doxorubicin, Immunomodulation
Introduction
Nanotechnology is a promising technology with potential advantages in treating and preventing disease. Among the metal nanoparticles, silver nanoparticles (AgNPs) are the most widely used nanoparticle in biomedical-related products because of their broad-spectrum antimicrobial activity. Recently, AgNPs have been reported to have antioxidant activity [1] and anti-inflammatory property [2]. Several studies have been reporting promising results of AgNPs for the treatment of inflammatory disease, such as for neuropathy in diabetic rats [3] and colitis [4].
Nanoparticles have an impact on the immune system and can either stimulate or suppress it. Nanoparticles may be recognized as foreign by immune cells, triggering an unintended immune response against them, potentially leading to toxicity within the body. The physical properties of nanoparticles, including their size, surface charge, and coating, play a crucial role in determining their compatibility with the immune system. Immune cells are likely involved in the disease’s pathophysiology as they receive signals from injured myocytes, endothelial cells, and cardiac progenitor cells [5].
Cardiotoxic exposures cause electrophysiological dysfunction or muscle damage to the heart. Typically, most acute cardiotoxic effects are reversible and thought to be the result of myocardial edema [6]. While chronic cardiotoxicity has been extensively studied, there is limited research on the manifestations of acute cardiotoxicity, and the development of secondary dilated cardiomyopathy is not fully understood. Doxorubicin (DOX) is a crucial and effective anticancer drug used in the treatment of many childhood cancer patients. However, its use carries potential risks for various complications in the short and long term. One of the most significant complications is the development of cardiotoxicity, which can lead to congestive heart failure [5].
Material and Methods
Chemicals
Doxorubicin was purchased from Sigma Chemical Co., St. Louis MO, USA.
Silver Nanoparticles (AgNPs): Dark gray powder (Particles Size Analysis {APS} 30-50nm) 1 g dissolved in saline, purchased from Sigma Aldrich (USA).
Experimental Animals
Sixty adult male albino rats weighing 198 ±11 g were used in this study. The animals were kept at standard housing facilities (24±1 ◦C, 45±5% humidity and 12 h light/dark cycle) They were supplied with standard laboratory chow and water ad libitum and left to acclimatize for 1 week before the experiments.
Experimental Design
Experimental rats were classified into six groups (n=10) as follows:
Group 1- Control (C): Rats received 1ml saline solution.
Group 2- AgNPs: Rats received AgNPs (4 mg/kg b.w) (NP1).
Group 3- AgNPs: Rats received AgNPs (8 mg/kg b.w) (NP2).
Group 4- Cardiotoxicity group (DOX): Rats received intraperitoneal single injection of DOX at dose of (30 mg/kg; i.p.) dissolved in saline solution, this dose is well proved to induce cardiotoxic effects.
Group 5 -Cardio-therapeutic1 (DOX+ NP1): Four days after DOX administration rats received AgNPs (4 mg/kg b.w).
Group 6- Cardio-therapeutic2 (DOX+ NP2): Four days after DOX administration rats received AgNPs (8 mg/kg b.w).
Treatments with AgNPs were given interperitoneally for 4 weeks.
Sample Collection
Blood was collected through retro-orbital puncture. The collected blood samples were left at a temperature of 24°C for 30 minutes, followed by centrifugation at 5000 r.p.m. for 20 minutes. The resulting serum was divided into multiple aliquots and stored at -20°C until analysis. For hematological analysis, heparinized blood was collected in EDTA-coated vials. The leukocytes count (WBC), lymphocytes count, and neutrophils count were determined from the whole blood using an automated hematological analyzer (Beckam Coulter, USA).
Serum and Heart Analysis
Several serum levels were measured in different experimental groups. These included anti-cardiolipin (ACL), troponin-T (TropT), CRP, albumin (Alb), LDH, CPK, AST, acid phosphatase activity (AP), TC, TG, total phospholipids (PL), and LDL-c. Additionally, the serum levels of tumor necrosis factor alpha (TNF-α), IL-6, NF-κB, IL-1β, WBC count, lymphocytes count, and neutrophils count were measured. A portion of each heart from all groups was taken and a 30% w/v homogenate was prepared in 0.9% buffered KCl (pH 7.4) to estimate GSH, MDA, SOD, and CAT.
Statistical Analysis
Data were shown as means ± standard error of mean (SEM), and a one-way analysis of variance (One-Way ANOVA) test. Differences were considered significant when P ≤ 0.05.
Ethical Approval
This study was approved by the ethics Committee of Sciences Academy of Experimental Research, AL-Mansoura, Egypt (2023-05-15, No. 44121).
Results
The administration of DOX significantly increased serum levels of cardiac markers, including troponin T (TropT), anti-cardiolipin (Acl), and C-reactive protein (CRP), while decreasing albumin levels (P ≤ 0.05), indicating cardiac damage. However, when AgNPs were administered to cardiotoxic rats, the cardiac damage caused by DOX was reduced. The administration of AgNPs as a therapeutic agent restored TropT, ACL, albumin, and CRP levels (Table 1).
Table (2) shows the effect of DOX and AgNPs on serum cardiac enzymes markers LDH, CPK, AP, and AST. DOX administration increased serum cardiac enzymes significantly (P ≤ 0.05). It was observed that administering AgNPs as therapeutic restore levels of serum enzymes near to normal levels. The rats treated with doxorubicin showed significantly higher levels of total cholesterol (TC), triglycerides (TG), total phospholipids (PL), and LDL-c compared to the control group (p≤ 0.05). However, the administration of silver nanoparticles (AgNPs) as a therapeutic intervention (G5, G6) improved these parameters and significantly reduced levels of TC and TG compared to the group experiencing cardiotoxicity (G4) (Figure 1). The findings indicated a connection between DOX-induced cardiotoxicity and a decrease in white blood cells count (WBCs), neutrophils, and lymphocytes. When compared to the cardiotoxic group (G4), the administration of AgNPs (G5, G6) resulted in an increase in leukocytes, lymphocytes, and neutrophils count. Treating cardiotoxic rats with AgNPs at either 4 mg/kg or 8 mg/kg effectively restored these counts in a dose-dependent manner (Figure 2). The administration of DOX caused a substantial increase in MDA levels (127.27%) and a significant decrease in GSH (-49.55%), SOD (-57.00%), and CAT (-44.96%) levels compared to the normal control group. However, when DOX-intoxicated rats were treated with AgNPs (4 mg/kg, 8 mg/kg), there was a significant improvement in MDA levels and a notable enhancement in GSH, SOD, and CAT activities compared to the DOX group (Table 3). The findings indicated that there were notable increases in levels of serum TNF-α, NF-κβ, IL-6, and IL-1β when DOX was administered, in comparison to the control group. Conversely, when cardiotoxic rats were treated with AgNPs at either 4 mg/kg or 8 mg/kg, there was a significant reduction in these inflammatory mediators in a dose-dependent manner. Moreover, the levels of proinflammatory cytokines in the serum were significantly lower in rats given the higher dose of AgNPs (8 mg/kg) compared to those given the lower dose of AgNPs (4 mg/kg) (Figure 3).
Discussion
Nanoparticles have become increasingly important in therapeutic applications in medicine due to their small size and large surface-to-volume ratio. Recent data has raised concerns about the use of silver nanoparticles (AgNPs) in therapeutics, as they have been found to be less toxic compared to other noble metal nanoparticles. Once inside the body, these nanoparticles encounter immune cells in the bloodstream, such as lymphocytes and granulocytes [7]. When tissue damage occurs and inflammatory stimuli and chemokines are released, neutrophils are attracted to these signals, leaving the circulation, and migrating to the site of damage. The number of neutrophils is related to the size of the infarction and to the development of heart failure [8]. Silver could interact with these immune cells and either stimulate or suppress them, leading to different pathological conditions [9].
Immune cells recognize silver nanomaterials as foreign particles, which can lead to an inflammatory response involving the activation of neutrophils and helper T cells. This activation results in the production of various cytokines, including tumor necrosis factor-α (TNF-α) and Interleukins (IL-1β, IL-6). These cytokines play a crucial role in the body’s natural defense against diseases and used in immunotherapies and vaccines. In a study conducted by Liu et al. [10], it was demonstrated that carefully designed silver nanomaterials effectively suppressed the recruitment of inflammatory cells to affected tissues, leading to the prevention of inflammation in mice.
Modulation of immune response may be stimulating or suppressive, but it is crucial to avoid excessive stimulation or suppression when using AgNPs in medical products. The effects of AgNPs on immune cells depend on their physicochemical properties and stability in biological environments, and thorough evaluation is necessary regardless of the intended purpose of the nanoparticles [11].
In this study, we examined the immunomodulatory effects of AgNPs at non-toxic concentrations. It is important to distinguish between immunotoxicity and immunomodulatory activity of nanomaterials, Previous studies have investigated the anti-inflammatory and antiproliferative effects of AgNPs by assessing cytokine expression using immunoassays with limited sensitivity and specificity [11]. The observed decrease in cytokine expression could potentially be attributed to the cytotoxic effects of AgNPs, particularly at higher concentrations. Notably, coadministration of AgNPs resulted in reduced expression of NF-κB, indicating that the formulation of AgNPs may have a mitigating effect on NF-κB-mediated inflammatory response [12].
In the present study, myocardial injury in rats is induced by doxorubicin (DOX) as evidenced by a notable rise in CPK and LDH activities. These elevated activities serve as indicators of leakage of cardiac enzymes. However, in the group treated with silver nanoparticles (AgNPs), CPK activity was significantly reduced compared to the DOX group, suggesting the protective effects of silver against myocardial damage.
In the study conducted by Nordgren and Wallace [13], it was observed that reactive oxygen species (ROS) can function as immune mediators and have an impact on different types of immune cells. When the levels of ROS are increased, immune cells can experience dysfunction, resulting in a state of immunosuppression. The development of cardiotoxicity because of doxorubicin treatment is a complex process that involves the regulation of multiple mechanisms of action [14]. The major mechanisms leading to cardiomyocyte cell death include the generation of reactive oxygen species (ROS) and nitrogen species that leads to protein and DNA damage and lipid peroxidation This leads to regulated or unregulated cell death apoptosis as well as necroptosis and the eventual release of inflammatory mediators [15].
In the present study, we demonstrated that doxorubicin (DOX) not only caused a significant disruption in the balance of redox status, but also induced lipid peroxidation, as evidenced by the elevated levels of systemic MDA in the heart. Additionally, this damage was accompanied by an inflammatory response, as indicated by the increased levels of IL-6 after DOX exposure. Previous research has also observed an increase in IL-6 in response to DOX [16]. Results of the current study showed a significant increase of MDA level after DOX administration. The protective role of AgNPs against oxidative stress-induced myocardial damage was observed through changes in MDA and SOD values. Co-administration of AgNPs resulted in a significant decrease in MDA levels and an increase in SOD levels. Previous studies have reported the pro-oxidant effects of AgNPs [17].
The role of inflammatory cytokines and chemokines in the development of myocardial dysfunction and cardiac remodeling is crucial. These versatile cytokines are increased in patients who have suffered a myocardial infarction as a response to myocardial injury. The initial stage of remodeling involves the secretion of TNFα, IL-1β, and IL-18, which promote inflammation. The subsequent stage is characterized by the release of anti-inflammatory cytokines. The activation of NF-κB, a major transcription factor in response to oxidative stress, has been established to contribute to cell proliferation and differentiation, thus playing a role in cardioprotective effects. In addition to its cardioprotective effects, AgNPs have also been implicated in early inflammatory responses through the activation of NF-κB, leading to the production of inflammatory cytokines such as TNFα, IL-1β, and IL-6. Previous studies have shown that the activation of NF-κB is crucial in the progression of cardiotoxicity induced by doxorubicin. [18]. In the current study, the administration of doxorubicin resulted in an elevation of NF-κB. However, it was found that co-treatment with AgNPs reduced the expression of NF-κB, indicating that AgNPs may have a mitigating effect on the inflammatory response mediated by NF-κB.
Cardiac troponins are the serum biomarkers of choice for monitoring potential drug-induced myocardial injury in both clinical and preclinical studies. In a study conducted by Lipshultz et al. [19], it was found that children treated with DOX experienced increased levels of cardiac troponin, even after the treatment had stopped. This suggests that there was damage to the heart and irreversible necrosis of cardiomyocytes.
The elevation of serum lipid levels caused by DOX toxicity is a clear indication of its well-known hyperlipidemic effect, as stated by Xiong et al. [20]. Our study observed notable increases in serum cholesterol and LDL-c levels, indicating that doxorubicin (DOX) hindered the breakdown of lipids. Consequently, our findings suggest that the detrimental effects of DOX, such as hyperlipidaemia, may contribute to the development of doxorubicin-induced heart failure, which negatively impacts heart function [21]. High levels of circulating cholesterol and its accumulation in heart tissue are well associated with cardiovascular damage [22]. Changes in lipid metabolism can affect cardiac function by altering the characteristics of the cardiac cell membrane [23].
The study revealed severe biochemical changes as well as oxidative damage in the cardiac tissue after the administration of DOX. Transaminases such as AST are liberated into the serum after extensive tissue injury. Because the heart muscle is rich in AST, it suggests that the increased level is an indicator of myocardial damage [24]. The result of the present study revealed that DOX intoxication caused a significant increase in acid phosphatase (AP) activity in heart tissues. Acid phosphatase activity on endothelial cells is responsible, in part, for the conversion of adenosine nucleotides to adenosine, a potent vasodilator and anti-inflammatory mediator that can protect tissues from the ischemic damage that results from injury. The cardiac biomarker enzyme LDH is extensively used in clinical practice as markers for the diagnosis of cardiac toxicity. Doxorubicin induces marked cardiotoxicity which was demonstrated by an increase in CPK and LDH activities. The magnitude of LDH activity in blood after myocardial injury reflects the extent of damage in its musculature. Increasing the serum levels of LDH have been indicated to cardiac tissue dysfunctions because these are normally located in the cytoplasm of cardiomyocytes and leakage occurs into the serum after cardiomyocytes damage [25].
Conclusion: The findings of this study suggest that silver nanoparticles (AgNPs) may have a therapeutic effect on the heart against cardiotoxicity induced by doxorubicin (DOX). AgNPs showed immunomodulatory potential as shown by an increase in blood leukocytes count, decrease in proinflammatory cytokines, and oxidative stress biomarkers.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: None
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Docea AO, Calina D, Buga AM, Zlatian O, Paoliello MMB, Mogosanu GD, et al. The effect of silver nanoparticles on antioxidant/pro-oxidant balance in a murine model. International Journal of Molecular Sciences. 2020;21(4):1233-49.
2. Prabakaran AS, Mani N. Anti-inflammatory activity of silver nanoparticles synthesized from Eichhornia crassipes: An in vitro study. Journal of Pharmacognosy and Phytochemistry. 2019;8(4):2556-8.
3. Alkhalaf MI, Hussein RH, Hamza A. Green synthesis of silver nanoparticles by Nigella sativa extract alleviates diabetic neuropathy through anti-inflammatory and antioxidant effects. Saudi Journal of Biological Sciences. 2020;27(9):2410-19.
4. Ige SF, Adeloye AA, Anifowose PE, Akinola OB, Badmus JA. Therapeutic potential of silver nanoparticles biosynthesized using aqueous extract of citrus sinensis peel on acetic acid-induced ulcerative colitis in male wistar rats. International Journal of BioLife Sciences. 2022;1(4):202-19.
5. Mancilla TR, Iskra B, Aune GJ. Doxorubicin-induced cardiomyopathy in children. Comprehensive Physiology. 2019;9(3):905-31.
6. Dulf PL, Mocan M, Coadă CA, Dulf DV, Moldovan R, Baldea I. et al. Doxorubicin-induced acute cardiotoxicity is associated with increased oxidative stress, autophagy, and inflammation in a murine model. Naunyn Schmiedebergs Arch Pharmacol. 2023;396(6):1105-1115.
7. Mills JA, Liu F, Jarrett TR, Fletcher NL, Thurecht KJ. Nanoparticle based medicines: approaches for evading and manipulating the mononuclear phagocyte system and potential for clinical translation. Biomaterials science. 2022;10(12):3029-53.
8. Sreejit G, Johnson J, Jaggers RM, Dahdah A, Murphy AJ, Hanssen NMJ, et al. Neutrophils in cardiovascular disease: warmongers, peacemakers, or both? Cardiovascular Research. 2022;118(12):2596-609.
9. Wei L, Lu J, Xu H, Patel A, Chen ZS, Chen G. Silver nanoparticles: synthesis, properties, and therapeutic applications. Drug discovery today. 2015;20(5):595-601.
10. Liu X, Gao P, Du J, Zhao X, Wong KK. Long-term anti-inflammatory efficacy in intestinal anastomosis in mice using silver nanoparticle-coated suture. Journal of Pediatric Surgery. 2017;52(12):2083-97.
11. Ninan N, Goswami N, Vasilev K. The impact of engineered silver nanomaterials on the immune system. Nanomaterials. 2020;10(5):967-93.
12. Azeem MN, Ahmed OM, Shaban M, Elsayed KN. In vitro antioxidant, anticancer, anti-inflammatory, anti-diabetic and anti-Alzheimer potentials of innovative macroalgae bio-capped silver nanoparticles. Environmental Science and Pollution Research. 2022;29(39):59930-47.
13. Nordgren KK, Wallace KB. Disruption of the Keap1/Nrf2-antioxidant response system after chronic doxorubicin exposure in vivo. Cardiovascular Toxicology. 2020;20(6):557-70.
14. Wenningmann N, Knapp M, Ande A, Vaidya TR, Ait-Oudhia S. Insights into doxorubicin-induced cardiotoxicity: molecular mechanisms, preventive strategies, and early monitoring. Molecular pharmacology. 2019;96(2):219-32.
15. Christidi E, Brunham LR. Regulated cell death pathways in doxorubicin-induced cardiotoxicity. Cell death & disease. 2021;12(4):339-53.
16. Jiang Y and Zhang Q. Catalpol ameliorates doxorubicininduced inflammation and oxidative stress in H9C2 cells through PPARγ activation. Experimental and Therapeutic Medicine. 2020;20(2):1003-11.
17. Dziendzikowska K, Wilczak J, Grodzicki W, Gromadzka-Ostrowska J, Węsierska M, Kruszewski M. Coating-dependent neurotoxicity of silver nanoparticles—an in vivo study on hippocampal oxidative stress and neurosteroids. International Journal of Molecular Sciences. 2022;23(3):1365-84.
18. Yarmohammadi F, Karbasforooshan H, Hayes AW, Karimi G. Inflammation suppression in doxorubicin-induced cardiotoxicity: Natural compounds as therapeutic options. Naunyn Schmiedebergs Arch Pharmacol. 2021;394(10):2003-11.
19. Lipshultz SE, Lipsitz SR, Sallan SE, Dalton VM, Mone SM, Gelber RD, et al. Chronic progressive cardiac dysfunction years after doxorubicin therapy for childhood acute lymphoblastic leukemia. Journal of clinical oncology. 2005;23(12):2629-36.
20. Xiong C, Wu YZ, Zhang Y, Wu ZX, Chen XY, Jiang P, et al. The protective effect of berberine on acute cardiomyopathy is associated with doxorubicin treatment. Oncology Letters. 2018;15(4):5721-29.
21. Sheibani M, Azizi Y, Shayan M, Nezamoleslami S, Eslami F, Farjoo MH, et al. Doxorubicin-induced cardiotoxicity: an overview on pre-clinical therapeutic approaches. Cardiovascular Toxicology. 2022;22(4):292-310. 92-310.
22. Klinnikova MG, Lushnikova EL, Koldysheva EV, Tolstikova TG, Sorokina IV, Yuzhik EI, et al. Cardiotoxic and dyslipidemic effects of doxorubicin and betulinic acid amide. Bulletin of experimental biology and medicine. 2016;162(2):277-82.
23. Zhang ZH, Yang CT, Su XR, Li YP, Zhang XJ, Wang SJ, et al. CCK1R2R-/-ameliorates myocardial damage caused by unpredictable stress via altering fatty acid metabolism. Stress. 2023;26(1):2254566-79.
24. Aldous SJ. Cardiac biomarkers in acute myocardial infarction. International journal of cardiology. 2013;164(3):282-94.
25. Ross JM, Öberg J, Brené S, Coppotelli G, Terzioglu M, Pernold K, et al. High brain lactate is a hallmark of aging and caused by a shift in the lactate dehydrogenase A/B ratio. Proceedings of the National Academy of Sciences. 2010;107(46):20087-92.
Download attachments: 10.4328.ACAM.22032
Nesrin I. Tarbiah, Fares K. Khalifa, Nuha A. Alkhattabi, Reem F. Ghazali, Reem Y. Alzahri, Sahar A. Alkhodair, Reham A. Shindi, Ahd A. Mansour. Immunomodulatory potential of silver nanoparticles and therapeutic effect against doxorubicin-induced-cardiotoxicity in rats. Ann Clin Anal Med 2024;15(3):169-175
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
Bloodstream infections: Epidemiology, risk factors, and resistance profiles in a university hospital: A five-year cross-sectional analysis
Meltem Karslioğlu 1, Aydın Deveci 2
1 Department of İnfections Diseases and Clinical Microbiology, Samsun Gazi State Hospital, 2 Department of İnfections Diseases and Clinical Microbiology, Faculty of Medicine, Ondokuz Mayıs Üniversity, Samsun, Turkey
DOI: 10.4328/ACAM.22039 Received: 2023-11-05 Accepted: 2023-12-11 Published Online: 2024-01-16 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):176-181
Corresponding Author: Meltem Karslioğlu, Department of İnfections Diseases and Clinical Microbiology, Samsun Gazi State Hospital, Samsun, Turkey. E-mail: celikmltm@hotmail.com P: +90 534 540 48 42 Corresponding Author ORCID ID: https://orcid.org/0000-0001-8490-3757
This study was approved by the Ethics Committee of Ondokuz Mayıs University, Faculty of Medicine (Date: 2015-05-14, No: OMÜ KAEK 2015/230)
Aim: Bloodstream infections present a significant healthcare challenge, causing substantial morbidity and mortality despite advancements in antimicrobial therapy. This study, conducted at the Infectious Diseases and Clinical Microbiology Clinic at Ondokuz Mayıs University (OMU), aimed to evaluate epidemiological characteristics, risk factors, infectious agents, and resistance profiles in hospitalized patients with bloodstream infections.
Material and Methods: This cross-sectional prospective analysis encompassed adult patients admitted to OMU between 2015 and 2019. Data included susceptibility tests, infection focus, treatments, and patient survival. Patients were categorized as having bacteremia or sepsis. Blood culture samples were collected with strict sterile procedures.
Results: Of the 100 patients, 47 had community-acquired infections, and 53 had nosocomial infections. The most common community-acquired focus was the urinary system (42.6%), and the most common nosocomial focus was intravenous catheters (43.4%). Escherichia coli (E. coli) was the most common causative microorganism, with 23% prevalence. Extended-spectrum beta-lactamase (ESBL) was detected in E. coli (34.7%) and multi-drug resistance in 47.8%. Klebsiella spp. exhibited ESBL (61.5%), multi-drug resistance (38.4%), carbapenem resistance (23%), and other resistances. Staphylococcus aureus had 28.5% methicillin resistance.
Discussion: This study offers vital insights into bloodstream infections, revealing their prevalence, causes, and resistance patterns. The challenge of drug-resistant organisms, especially ESBL and carbapenem-resistant bacteria, emphasizes the need for tailored treatment strategies and collaborative efforts. The increasing prevalence of MRSA and VRE underscores the importance of prudent antibiotic use and rigorous infection control. In conclusion, this study calls for a collective approach to address evolving risks in bloodstream infections, improving patient outcomes and public health.
Keywords: Bloodstream Infections, Community Acquired Infections, Nosocomial Infections, Antibiotic Resistance
Introduction
Bloodstream infections, despite aggressive broad-spectrum antimicrobial therapies and supportive care, remain a formidable challenge, contributing significantly to morbidity and mortality rates[1]. According to the Centers for Disease Control and Prevention (CDC), rising rates of antibiotic resistance add a layer of complexity to the management of these life-threatening infections (Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4378521/). It has therefore become more important to rigorously study the infectious agents that cause these conditions and to develop stronger measures for accurate diagnosis and effective treatment.
While the spectrum of pathogens causing bloodstream infections may fluctuate over time, bacteria continue to dominate as the primary culprits [2,3]. The stealthy presence of viruses often eludes diagnosis, further complicating the scenario [4]. Gram-negative bacteria typically take center stage, but the surge in gram-positive bacterial infections, driven by the increasing prevalence of interventional procedures, warrants close attention [5]. Additionally, the past decade has witnessed a notable upsurge in bloodstream infections caused by fungal agents, underscoring a shifting landscape [6]. Furthermore, it is crucial to recognize that the identities of isolated infectious agents and their susceptibility profiles exhibit considerable variation, contingent upon the demographic attributes of patients and the specific treatment regimens employed [7]. As a result, maintaining a watchful eye on these infections is of utmost importance, as it paves the way for the implementation of tailored empirical treatments aligned with the unique profiles of individual patients [8].
This study, conducted at the Infectious Diseases and Clinical Microbiology Clinic at the Ondokuz Mayıs University, sets out with a clear aim: to comprehensively assess the epidemiological characteristics, risk factors, infectious agents, and their resistance profiles in patients hospitalized with bloodstream infections. The overarching objective is to furnish a comprehensive understanding of these infections, offering insights into their prevalence, etiology, and the pressing matter of resistance. The findings from this research promise to significantly inform and enhance medical practices in this vital field of healthcare.
Material and Methods
This study presents the results of research conducted within the framework of the thesis titled ‘Causative Agents and Resistance Issues in Community-Acquired and Nosocomial Sepsis.
Study Design: This cross-sectional prospective analysis encompassed adult patients (age > 18 years) admitted to Ondokuz Mayıs University’s Infectious Diseases and Clinical Microbiology service between 2015 and 2019. The study group included 100 patients, comprising 45 males and 55 females from diverse age groups. Patients were included based on the presence of causative microbial growth in their blood cultures.
Data Recording: Susceptibility tests were conducted on the causative microorganisms isolated from blood cultures, and the infection focus was documented. Coagulase-negative staphylococci that grew only once were not considered causative unless subsequent growth was consistent with clinical condition and similar sensitivity. For catheter-related infections, microorganisms exhibiting the same causative agent and sensitivity were regarded as significant if they concurrently grew in blood culture. Detailed records of antibiotic and supportive treatments, as well as patient survival status, were maintained. Day 0 was defined as the day when microorganisms were detected or growth signals observed, followed by comprehensive systemic evaluations.
Classification: Patients with microbial growth in blood cultures were classified as having bacteremia, while those with bacteremia and a SIRS score of 2 or higher were categorized as sepsis cases. Septic shock was defined for sepsis patients exhibiting hypotension requiring vasopressors to maintain a mean arterial pressure of ≥65 mmHg and serum lactate levels exceeding 2 mmol/L (18 mg/dl).
Blood Culture Collection: Blood culture samples were obtained by drawing blood from at least two distinct veins, with a 20-minute interval between samples, prior to initiating antibiotic therapy to prevent treatment delay. A rigorous cleansing process with 70% alcohol and 10% povidone iodine was performed at the puncture site before blood sample collection. A minimum of 10 ml of blood was collected. To maintain sterility, the rubber stopper of the blood culture bottle was disinfected with alcohol after removing the plastic cap, placed into the blood culture bottle, and wiped with alcohol once more.
Microbiological Analysis: Blood culture bottle growth was monitored using the automated BD BACTEC™ system (Becton Dickinson, Sparks, MD, USA). Bacterial identification was conducted with the VITEK®MS system (bioMérieux, France), and antibiotic susceptibility tests were carried out with the VITEK®2COMPACT device (bioMérieux, France). Extended-spectrum beta-lactamase (ESBL) identification was confirmed via double-disc synergy testing, following EUCAST standards for antibiotic susceptibility tests and ESBL enzyme identification.
Data Analysis
Statistical Analysis: Statistical analyses were performed using IBM SPSS 22 software. Sociodemographic characteristics of the patients were subjected to frequency analysis. Independent two-sample t-tests were used for normally distributed and binary variables, while non-normally distributed binary variables were analyzed with the Mann-Whitney U test. The Friedman test was applied for non-normally distributed dependent groups with more than two repetitions. Qualitative variable relationships were assessed using the Pearson Chi-Square test. All statistical tests were conducted at a 95% confidence level.
Ethical Approval
This study was approved by the Ethics Committee of Ondokuz Mayıs University, Faculty of Medicine (Date:14.05.2015 , No: OMÜ KAEK 2015/230).
Results
Out of 100 patients, 45% of patients were females (n=45), and 55% were males (n=55), with an average age of 62.93 ± 15.84 years. Among them, 47% (n=47) presented with community-acquired infections, while 53% (n=53) had nosocomial infections. The mean age for patients with community-acquired infections was 62.19 ± 17.62 years, and for those with nosocomial infections, it was 63.58 ± 14.22 years. There was no statistically significant age difference between these two groups (p=0.663). When examining the comorbidities of the patients, it was found that in the community-acquired infections group, 12 patients (25.5%) had diabetes mellitus (DM), whereas in the nosocomial infections group, 16 patients (30.2%) had DM. There was no statistically significant difference in terms of DM between the groups (p>0.05). However, in terms of chronic kidney disease (CKD), 7 patients (14.9%) in the community-acquired infections group had CKD, while 22 patients (41.5%) in the nosocomial infections group had CKD. A statistically significant difference was observed in CKD between the two groups (p=0.007). There were no statistically significant differences between the groups regarding other comorbidities, including chronic liver disease (CLD), congestive heart failure (CHF), chronic obstructive pulmonary disease (COPD), immunosuppressive therapy (IS therapy), and hypertension (HTN) (p>0.05).
In our study, we investigated the spectrum of causative microorganisms in bloodstream infections, focusing on their differentiation between nosocomial and community-acquired cases among 100 patients. Notably, Escherichia coli emerged as the predominant pathogen, responsible for 23% of all cases. However, the distribution of causative microorganisms revealed striking disparities between nosocomial and community-acquired infections. Within nosocomial infections (53% of the cases), Escherichia coli remained a significant contributor, causing 13.2% of these cases. Staphylococcus aureus was also prevalent, responsible for 11.3% of the nosocomial infections. On the other hand, community-acquired infections saw an even higher incidence of Escherichia coli, contributing to 34% of the cases, along with 17% attributed to Staphylococcus aureus. Furthermore, skin and soft tissue infections played a substantial role in community-acquired cases, accounting for 19.1% (Table 1).
As depicted in Table 2, among patients with community-acquired infections, the most common primary sources of infection were the urinary tract (42.6%, n=20) and skin and soft tissue (19.1%, n=9). Conversely, for patients with nosocomial infections, intravenous catheters (43.4%, n=23) and urinary tract (26.4%, n=14) stood out as the predominant sources of infection.
In our comprehensive analysis of bloodstream infections, we observed distinctive resistance patterns among Escherichia coli, Klebsiella spp., and Acinetobacter spp., categorized by nosocomial and community-based infections. Among Escherichia coli cases, 34.7% (n=8) were ESBL-positive, while 65.3% (n=15) were ESBL-negative. Additionally, 47.8% (n=11) exhibited multi-drug resistance (MDR), and the remaining 52.2% (n=12) were MDR-negative, notably showing no carbapenem resistance. Klebsiella spp. presented a higher prevalence of ESBL-positive cases, with 61.5% (n=8), and 38.5% (n=5) were ESBL-negative. Among them, 38.4% (n=5) showed MDR, while 61.6% (n=8) were MDR-negative. Notably, 23.1% (n=3) displayed carbapenem resistance, and 7.6% (n=1) exhibited extreme drug resistance (XDR) and pan drug resistance (PDR). In the case of Acinetobacter spp., 25% (n=1) showed MDR, while the remaining 75% (n=3) were MDR-negative, and all cases displayed carbapenem resistance. Furthermore, when considering nosocomial and community-based infections, ESBL-positive cases were equally distributed, with 50% (n=13) in each group. Carbapenem resistance was observed in 46.7% (n=7) of community-based Klebsiella spp. infections, 40% (n=6) in Acinetobacter spp., and 13.3% (n=2) in Providencia rettgeri (Table 3).
Discussion
Bloodstream infections, affecting over 30 million people worldwide, remain a significant global health challenge, causing substantial morbidity and mortality despite ongoing medical advancements [9]. With increasing life expectancy, individuals are increasingly exposed to the risk of infection. Westphal et al.’s study, akin to ours, found that both community-acquired and nosocomial infections predominantly affect individuals aged 60 or older. This higher average age can be attributed to the greater prevalence of chronic diseases and age-related weakening of the immune system, which elevates the susceptibility to infections. Furthermore, advanced age often necessitates more invasive medical interventions, which, in turn, heightens the risk of infection [10].
In another retrospective study by Rhee et al., intra-abdominal infections were identified as the most common source (20.6%) of nosocomial infections, followed by pneumonia, urinary tract infections, and less common soft tissue infections [11]. Similarly, a retrospective analysis of community-acquired bloodstream infections in our country revealed urinary tract infections (45%), pneumonia (18%), intra-abdominal infections (9.6%), and skin and soft tissue infections (5%) as the primary sources [12]. It is noteworthy that our study pinpointed urinary tract infections and skin and soft tissue infections as predominant sources in community-acquired cases. The differences in infection source rankings can be attributed to our comprehensive approach, which involved managing patients admitted to the infectious diseases service, as opposed to cases managed by different clinics and consultation services for lung and abdominal infections. This highlights the critical need for tailored treatment strategies and interdisciplinary collaboration to address these infections.
The Extended Prevalence of Infection in Intensive Care (EPIC) study, utilizing the point prevalence method, reported lung infections accounting for 64% of cases in intensive care units, followed by abdominal infections (20%), vascular catheter-related infections (15%), and genitourinary system infections (14%) [13]. Pneumonia was the most frequent infection, followed by vascular catheter-related and urinary system infections. Conversely, our study, focusing on patients admitted to the infectious diseases service, highlighted the predominance of vascular catheter-related infections since patients diagnosed with pneumonia were typically managed by pulmonary diseases clinics or anesthesia/internal medicine intensive care units. The growing number of dialysis patients, requiring vascular catheters, emphasized the prominence of vascular catheter-related infections in our study, ultimately making them the leading cause of hospital-acquired infections.
Bacterial infection rates in these patients were found to be 30-66.4%. In a study conducted in our country, culture growth was found in 46.5% of the total patients, 45.5% of the patients hospitalized in the chest diseases service with chronic obstructive pulmonary disease (COPD) attack and 52.5% of the patients hospitalized in the intensive care unit [14]. Similarly, our study significantly increases the risk of COPD, especially community-acquired infections.
When analyzing the microbial agents responsible for bloodstream infections, a study conducted in our country reported Klebsiella pneumoniae and Escherichia coli as the most common gram-negative pathogens in blood cultures [15]. Meanwhile, Staphylococcus aureus was the predominant gram-positive pathogen. In our study, Escherichia coli was the most common gram-negative agent, but when distinguishing between community-acquired and nosocomial infections, Klebsiella spp. became the primary pathogen in nosocomial cases. A similar study focusing on community-acquired infections in the elderly also identified Escherichia coli as the most common pathogen, aligning with our findings [16]. The similarities in results are likely due to the majority of patients in our study being managed in the intensive care unit.
The emergence of Extended-Spectrum Beta-Lactamases (ESBL) in E. coli and Klebsiella species, attributed to their ability to spread among strains, is a notable concern. ESBL rates can vary between outpatients and inpatients, the types of samples from which strains are isolated, whether the agent is nosocomial, and over the years [17]. Although our study didn’t show a statistically significant difference, the rate of ESBL in community-acquired infections was 33%, while it nearly doubled in hospital-acquired infections, potentially indicating a more resistant profile in the latter.
Carbapenem resistance was not detected in E. coli strains from two separate studies involving urinary tract infections in our country, aligning with our findings [18]. This may be due to the limited use of carbapenems, especially in outpatients, due to their parenteral administration
In ESBL-positive K. pneumoniae and K. oxytoca strains, carbapenems are highly effective, particularly after amikacin. However, the overuse of carbapenems without adhering to rational antibiotic use policies can lead to resistance development. Studies conducted in our country by Görgec et al. and Copur et al. found that ESBL-positive K. pneumoniae isolates exhibited resistance rates of approximately 5% for imipenem and meropenem [19,20]. In our study, we also detected carbapenem resistance, with a rate of 7%, which aligns with previous research. While carbapenem resistance remains lower than ESBL resistance, it’s noteworthy that it has increased compared to the past. This emphasizes the importance of considering this trend, especially when making empirical treatment choices [21]. Both studies identified a history of hospitalization as a risk factor for carbapenem resistance in Acinetobacter spp. strains. In our study, Acinetobacter spp. was one of the two carbapenem-resistant agents. This highlights the significance of rational antibiotic use, as carbapenem-resistant Acinetobacter spp. strains are increasingly observed in ward patients, extending beyond the confines of intensive care units.
The rate of antibiotic use is increasing, and this increase brings along drug resistance. Therefore, antibiotic treatment strategies need to be reconsidered. In our study, it was determined that a history of antibiotic use was a risk factor for the development of infection with both ESBL-positive and other multi-drug resistant agents. The history of hospitalization, which is frequently mentioned in the literature, was also examined by us, but although ESBL is more common in patients with a history of hospitalization, no statistically significant difference was found in terms of resistance development.
It draws attention with different studies that resistance to methicillin in staphylococci has increased over the years: In different studies, MRSA rates were found to be 31.7% in 1988, 35.3% in 1992, 31.6% in 1994, 36.1% in 1998, 60.2% in 2000, and 64% in 2001. Grundmann et al.[22] On the other hand, in the study they conducted between 1999 and 2002, they found the rate of MRSA to be 5-20% [23]. The low is quite remarkable. In our study, this rate was determined as 28.5%, and although it seems to be an average value according to sample studies, it is quite remarkable that this rate was much lower in the past years.
In a study conducted in our country for Enterococcus strains, penicillin resistance was found to be 48%, ampicillin resistance was 43% in samples taken from outpatients, and vancomycin and teicoplanin resistance were not detected. Penicillin resistance was found to be 84%, ampicillin resistance to 70%, and vancomycin and teicoplanin resistance to 5% in the samples sent from hospitalized patients [24]. Although we had lower ampicillin resistance compared to the sample study, vancomycin resistance was much higher with 20%. Today, patients are colonized and infected with vancomycin-resistant enterococci in many countries. These infections are increasing due to non-compliance with rational antibiotic use policies. Therefore, it is important to detect resistant strains and to know their resistance rates.
Conclusion:
In summary, this study underscores the growing challenge of bloodstream infections, especially those caused by drug-resistant organisms. To combat this threat, we must prioritize rational antibiotic use, enhance infection control measures, and foster interdisciplinary collaboration. Addressing these issues collectively is essential for improving patient outcomes and safeguarding public health.
Limitations
This study has some limitations. Firstly, the fact that the research was conducted at a single medical center and had a limited sample size may limit the generalizability of the results. Additionally, epidemiological changes during the data collection period or changes in hospital practices could impact the findings. The retrospective nature of the data used in the study may increase the possibility of data gaps or inaccuracies. Finally, relying on data from only one medical center may lead to different results in different geographical regions or healthcare systems. Therefore, conducting similar studies in different centers with larger sample groups is crucial for obtaining more reliable results.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: None
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Afzal S. GBD 2019 Antimicrobial Resistance Collaborators. Global mortality associated with 33 bacterial pathogens in 2019: a systematic analysis for the Global Burden Of Disease Study 2019. Lancet. 2022;400(10369):2221-2248.
2. Martinez RM, Wolk DM. Bloodstream Infections. Microbiol Spectr. 2016;4(4):10-31.
3. Bandy A, Almaeen AH. Pathogenic spectrum of blood stream infections and resistance pattern in Gram-negative bacteria from Aljouf region of Saudi Arabia. PLoS One. 2020;15(6):1-14.
4. Louten J. Detection and diagnosis of viral infections. Essential Human Virology. 2016 (1):111–32.
5. Oliveira J, Reygaert WC. Gram-Negative Bacteria. StatPearls. Treasure Island (FL): StatPearls Publishing; 2023.p.87-93.
6. Mamali V, Siopi M, Charpantidis S, Samonis G, Tsakris A, Vrioni G, et al. Increasing Incidence and Shifting Epidemiology of Candidemia in Greece: Results from the First Nationwide 10-Year Survey. J Fungi (Basel). 2022;8(2):116-117.
7. van Seventer JM, Hochberg NS. Principles of Infectious Diseases: Transmission, Diagnosis, Prevention, and Control. International Encyclopedia of Public Health. 2017 (1):22-39.
8. Resman F. Antimicrobial stewardship programs; a two-part narrative review of step-wise design and issues of controversy Part I: step-wise design of an antimicrobial stewardship program. Ther Adv Infect Dis. 2020;7:1-26.
9. Laupland KB, Church DL. Population-based epidemiology and microbiology of community-onset bloodstream infections. Clin Microbiol Rev. 2014;27(4):647-664.
10. Westphal GA, Pereira AB, Fachin SM, Barreto ACC, Bornschein ACGJ, Caldeira Filho M, et al. Characteristics and outcomes of patients with community-acquired and hospital-acquired sepsis. Características e desfechos de pacientes com sepse adquirida na comunidade e no hospital. Rev Bras Ter Intensiva. 2019;31(1):71-78.
11. Rhee C, Wang R, Zhang Z, Fram D, Kadri SS, Klompas M; CDC Prevention Epicenters Program. Epidemiology of Hospital-Onset Versus Community-Onset Sepsis in U.S. Hospitals and Association With Mortality: A Retrospective Analysis Using Electronic Clinical Data. Crit Care Med. 2019;47(9):1169-1176.
12. Güler H, Akalın H, Heper Y, Yılmaz E, Sinirtaş M, Öztürk Ç, et al. Toplum kökenli sepsis: 125 olgunun retrospektif değerlendirmesi [Community-acquired sepsis: Retrospective evaluation of 125 cases]. Flora İnfeksiyon Hastalıkları ve Klinik Mikrobiyoloji Dergisi. 2010;15(1):11–15.
13. Vincent JL, Rello J, Marshall J, Silva E, Anzueto A, Martin CD, et al.; EPIC II Group of Investigators. International study of the prevalence and outcomes of infection in intensive care units. JAMA. 2009 Dec 2;302(21):2323-2329.
14. Behçet M, Avcioğlu F, Özsari E, Tuğ T, Kurtoğlu MG, KOAH Akut Atak Nedeniyle yatan hastalarda bakteriyel enfeksiyon etkenleri, antibiyotik direnci ve komorbiditenin araştırılması [investigation of bacterial infection agents, antibiotic resistance and comorbidity in patients hospitalized for COPD acute attack]. DÜ Sağlık Bil Enst Derg. 2020;10(1):6–11.
15. Aydin M, Kasikcioglu C, Nargiz-Kosucu S, Timurkaynak F, and Arslan, H. Bloodstream Infections: Etiologic Agents and Their Antibiotic Resistance Rates [Kan Dolasimi Infeksiyonu Etkenleri ve Antibiyotik Direnc Oranlari]. KLIMIK Journal, 2016;29(2): 82-100.
16. Roubaud Baudron C, Panhard X, Clermont O, Mentré F, Fantin B, Denamur E, et al. Escherichia coli bacteraemia in adults: age-related differences in clinical and bacteriological characteristics, and outcome. Epidemiol Infect. 2014;142(12):2672-2683.
17. Pekintürk NS, Akgüneş A. Yatan hastalardan izole edilen Escherichia coli ve Klebsiella spp. suşlarında genişlemiş spektrumlu beta-laktamaz üretimi ve antibiyotik direnç oranları: 2011-2015 verileri [Extended-spectrum beta-lactamase production and antibiotic resistance rates in Escherichia coli and Klebsiella spp. isolated from inpatients: 2011-2015 data]. Turk Hij Den Biyol Derg. 2017;74 8(3):221–228.
18. Güneş H, Donma M, Nalbantoğlu B, Aydın M, Kaya A, Topcu B. Namık kemal üniversitesi araştırma ve uygulama hastanesi’ne başvuran çocuklarda idrar örneklerinden izole edilen etkenler ve antibiyotik direnç durumları [agents isolated from urine samples of children attended the namik kemal university research and practice hospital and the rate of antibiotic resistance]. CMJ. 2013;35 (1):1–8.
19. Çopur Çiçek A, Şentürk Köksal Z, Ertürk A, Köksal E. Microorganisms isolated from blood cultures during the period of one year at the 82nd Year Rize State Hospital and their susceptibility to antibiotics. Turkish Bulletin of Hygiene and Experimental Biology. 2011;68(4):175–184.
20. Görgeç S, Kuzucu Ç, Yetkin F, Ersoy Y. Efficiency of Tigecycline and Other Antimicrobials to Extended Spectrum Beta Lactamase Producing Gram Negative Bacteria in Blood Culture Isolates and Detection in Activity of Carbapenemases. annalsmedres 2021;18(2):106-110.
21. van Duin D, Paterson DL. Multidrug-Resistant Bacteria in the Community: An Update. Infect Dis Clin North Am. 2020;34(4):709-722.
22. Grundmann H, Aires-de-Sousa M, Boyce J, Tiemersma E. Emergence and resurgence of meticillin-resistant Staphylococcus aureus as a public-health threat. Lancet. 2006;368(9538):874-885.
23. Dündar D, Tamer SG. Klinik örneklerden izole edilen staphylococcus aureus suşlarının antimikrobiyal duyarlılıkları: Üç yıllık değerlendirme [Antimicrobial susceptibility of staphylococcus aureus strains isolated from clinical specimens: A three-year evaluation]. ANKEM Dergisi. 2009;23(1):8–12.
24. Güçkan R. , Elmas A. , Tilgel S. , Yüksel G. Çeşitli Klinik Örneklerden İzole Edilen Enterokok Suşlarının Antibiyotik Duyarlılıkları [Antibiotic Susceptibilities of Enterococci Strains Isolated from Various Clinical Specimens]. International Journal of Basic and Clinical Medicine. 2013;1(2):74-77.
Download attachments: 10.4328.ACAM.22039
Meltem Karslioğlu, Aydın Deveci. Bloodstream infections: Epidemiology, risk factors, and resistance profiles in a university hospital: A five-year cross-sectional analysis. Ann Clin Anal Med 2024;15(3):176-181
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
Potential antiviral drug resistance mutations in patients with treatment-naive chronic hepatitis B
Bulent Cakal 1, Bilger Cavus 2, Mehves Poda 3, Alp Atasoy 2, Mesut Bulakcı 4, Mine Gulluoglu 5, Filiz Akyuz 2
1 Department of Medical Microbiology, Faculty of Medicine, 2 Department of Internal Medicine, Division of Gastroenterohepatology, Faculty of Medicine, 3 Department of Genetics, Aziz Sancar Institute for Experimental Medical Research, 4 Department of Radiology, Faculty of Medicine, 5 Department of Pathology, Faculty of Medicine, Istanbul University, Istanbul, Turkey
DOI: 10.4328/ACAM.22047 Received: 2023-11-16 Accepted: 2024-01-01 Published Online: 2024-01-10 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):182-187
Corresponding Author: Bülent Çakal, Department of Medical Microbiology, Istanbul Faculty of Medicine, Istanbul University, Topkapı Mahallesi, Turgut Özal Caddesi, No: 118, 34093, Fatih, İstanbul, Turkey. E-mail: bulentcakal@yahoo.com P: +90 532 726 64 54 Corresponding Author ORCID ID: https://orcid.org/0000-0002-1254-844X
This study was approved by the Ethics Committee of Istanbul Medical School Ethics Committee at Istanbul University (Date: 2018-06-12, No: 895)
Aim: The prevalence and clinical effects of mutations in the reverse transcriptase (RT) region of the Hepatitis B Virus (HBV) before antiviral treatment in patients with chronic Hepatitis B (CHB) are not clear. In this study, the aim was to investigate the prevalence, characteristics, antiviral drug resistance, and clinical effects of mutations in the RT region of the HBV polymerase gene in patients with treatment-naive CHB.
Material and Methods: The study included 102 treatment-naïve patients who underwent liver biopsy due to chronic hepatitis B disease. The gene region encoding HBV RT enzyme was amplified by PCR and Sanger sequencing was performed.
Results: The genotype of all patients with CHB was defined as HBV/D. In the study, among the 102 included patients, 82 (80.39%) had a total of 35 types of mutations associated with the 42 previously identified Nucleos(t)ide analogs (NAs) resistance (NAr) mutation positions in the HBV RT region. In a total of 5 patients (4.90%), primary and secondary drug resistance were detected. In this study, 22 new mutations were identified in 13 out of 31 RT positions associated with NAr. No statistically significant differences were observed in terms of liver enzymes and histologies, as well as virological features including HBV DNA levels, between patients with detected NAr mutations and those without.
Discussion: Although the prevalence of amino acid (AA) variations in the HBV RT region is high in patients with HBV/D genotype, primary and secondary antiviral resistance associated with NAs is rare. Effects of NAr mutations on the progression of HBV-related diseases are also quite limited.
Keywords: Hepatitis B Virus, Chronic Hepatitis B, Treatment-Naive, Mutations, Polymerase Reverse Transcriptase.
Introduction
Chronic hepatitis B virus (HBV) infections continue to be a significant clinical problem worldwide. [1]. Currently, there is no effective treatment for the estimated 296 million individuals worldwide who are carriers of chronic HBV. It is estimated that approximately 820,000 people die from HBV-related liver diseases each year [2].
HBV replication occurs through the reverse transcription of viral pregenomic RNA. In this respect, HBV Polymerase/reverse transcriptase (Pol/RT) is a critically important enzyme for viral replication and serves as an antiviral target for the inhibition of HBV biosynthesis. [3]. Today, antiviral treatment strategies designed to achieve prolonged suppression of viral replication by targeting HBV RT include clinical use of nucleotide analogs (NAs) such as lamivudine (LMV), adefovir dipivoxil (ADV), telbivudine (LdT), entecavir (ETV), and tenofovir (TDF) for the treatment of patients with chronic hepatitis B (CHB) [4].
However, long-term treatments and mutations arising from the lack of proofreading ability of the HBV RT enzyme can lead to the development of antiviral drug resistance. Antiviral drug resistance caused by RT mutations can limit the clinical efficacy of treatment, leading to treatment failure and an increase in the progression of the disease. Moreover, mutations arising from the partially overlapping genomic structure of HBV’s surface antigen (HBsAg), encoded by the HBV surface (S) gene, with the RT can affect the antigenicity, immune recognition, virulence, and replication capacity of HBV [5].
In patients with CHB, 42 potential nucleotide analog resistance (NAr) mutation positions have been identified in the HBV RT region prior to treatment. However, the prevalence of preexisting mutations in the HBV polymerase gene’s RT region and their impact on antiviral treatment in treatment-naive patients with CHB is not yet clear [6, 7].
Therefore, the objective of this study is to determine the prevalence of preexisting mutations in the HBV RT region in treatment-naive patients with chronic hepatitis B (CHB) and to examine the potential and clinical effects of these mutations on antiviral treatment.
Material and Methods
This study was supported by the Scientific Research Projects Coordination Unit of Istanbul University with the project number TSA-2023-39107. All patients included in this study gave informed consent for participation.
Patients:
This study included 102 patients followed by the Gastroenterohepatology clinic of Istanbul University Istanbul Faculty of Medicine due to CHB disease. Liver biopsy procedures were performed at the Department of Internal Medicine, Division of Gastroenterohepatology, and the Interventional Radiology Unit of the Department of Radiology at Istanbul Faculty of Medicine.
Amplification of HBV Pol/RT gene:
Total genomic DNA samples, isolated from liver biopsy specimens of patients using a commercial kit (QIAamp DNA Mini kit, Qiagen GmbH, Hilden, Germany), were amplified by PCR method using HBV Pol/RT gene-specific primers (Table 1). To amplify the target region, a PCR setup (25 µl) was constructed, containing 12.5 µl 2X PCR master mix (HS Prime Taq Premix; GeNet Bio) and external primers (200ng/µl) for amplification of 5µl DNA extract. PCR conditions were as follows: 95°C for 10 min; 35 cycles of 95°C for 30 seconds, 55-60°C for 1 min and 72°C for 1 min; then 72°C for 10 min.
Sequencing of HBV Pol/RT gene:
PCR products detected to have bands of the desired length were purified for sequencing. The sequencing of the purified amplification products was carried out using amplification primers in a bidirectional manner through a genetic analyzer (Beckman Coulter CEQ 8000).
The nucleotide sequences of the HBV Pol/RT gene obtained after the sequencing process were edited using the Chromas program. The confirmation and bioinformatic analyses of the edited sequences were performed on the NCBI website (http://blast.ncbi.nlm.nih.gov/Blast.cgi). In determining the HBV genotypes, the NCBI website containing HBV reference sequences (www.ncbi.nlm.nih.gov/projects/genotyping) was used.
To identify mutations in the HBV Pol/RT gene, the Geno2pheno tool of the Max Planck Institute Informatik (https://hbv.geno2pheno.org/index.php) and the HIV-grade HBV Drug Resistance Interpretation program (http://www.hiv-grade.de/hbv grade/deployed/grade) were used.
The demographic, clinical, pathological, and laboratory data of the patients were obtained from patient files or, when necessary, from the hospital’s electronic records.
Statistical analysis
Statistical analyses were conducted using the SPSS software (version 25.0, SPSS Inc., Chicago, IL). The results were presented as means and standard deviations. The Chi-square test and/or Fisher test were used for categorical variables, and Mann-Whitney U and One-Way ANOVA tests were used to compare non-categorical variables. P< 0.05 was considered statistically significant. This study was approved by the Ethics Committee of Istanbul Medical School Ethics Committee at Istanbul University (Date: 2018.06.12, No: 895).
Results
Patients’ characteristics
The data regarding the demographic, clinical, histological, and laboratory results of the patients included in this study are summarized in Table 2. The study included 51 female and 51 male patients with chronic hepatitis B (CHB), with an average age of 41.7±13. The mean necroinflammatory activity scores and fibrosis stage levels of the patients with chronic hepatitis B included in the study were determined as 5 (0-17) and 2(0-6), respectively. The mean serum HBV DNA levels for patients with chronic hepatitis B (CHB) were Log10 5.31 (1.56 – >8.23).
Mutations on HBV Pol/RT gen of patients with CHB
The genotype of all isolated HBVs from patients with CHB was determined to be HBV D in 100% of cases.
Out of the 102 patients included in the study, at least one of the previously identified 42 NAr mutation positions in the RT region was detected in 82 (80.39%) individuals. In this study, 35 types of mutations were identified in 21 of 42 (50%) previously identified potential mutation positions for the RT region of the HBV polymerase gene (Tablo 3).
In this study, primary and secondary drug resistance were detected in a total of 5 patients (4.90%), with 4 patients (3.92%) showing resistance at the rtA194 position associated with TDF, and 1 patient (0.98%) at the rtV173 position (Tablo 3). No mutations associated with the previously identified primary and secondary drug resistance at 11 HBV RT positions were found in any of the patients, except for the two mentioned above.
In our study, 15 out of the previously identified 26 assumed NAr-associated mutation positions in the HBV RT region (57.7%) were detected. Nine out of these 15 RT positions were found to have 10 assumed mutations as described by Liu et al [6]. Additionally, in this study, 16 new mutations were identified at the 10 RT positions, different from the assumed mutations defined by Liu et al. The Y54H mutation, identified in 16 patients (15.68%), was the most common variation. (Tablo 3).
In this study, 5 out of the 6 previously identified pre-treatment RT mutation positions in the HBV RT region (83.3%) were detected. The pre-treatment mutation R242A, as defined by Liu and colleagues, was identified at only one RT position and in one patient in this study. Also, in this study, 7 new mutations were identified at 4 RT positions, different from the pre-treatment mutations defined by Liu et al, including N139T. Ultimately, in this study, 22 new mutations were identified in 13 of the 31 RT positions, (Tablo 3).
Thirteen out of the 21 amino acid positions where mutations were detected were genotype-associated polymorphic sites. In this study including treatment-naive patients, the amino acid (AA) positions rt38, rt53, rt54, rt124, rtH126, rt134, rt139, rt166, rt191, rt214, rt215, rt238, and rt256 in the HBV RT region were identified as the most polymorphic 13 (AA) positions. Among these, rtH124, detected in 41 patients (40.19%), was the most polymorphic amino acid position. The mutations detected in the functional domains (G, F, A, B, C, D, and E) and interdomains (F-A, A-B, B-C, C-D, and D-E) of patients in the HBV RT region, along with their prevalence, are showed in Table 4 in this study. Although mutations were detected at all sites, the most frequently mutated domain was identified as the A-B interdomain. The amino acid positions detected in this interdomain were R110, L115, N118, F122, H126, T128, S135, S137, N139, and Q149. H126R, T128I, and N139T were the most frequently observed variations in this domain. The C-D interdomain also contained the most variations with mutations at amino acid positions S213, V214, Q215, F221, and L229.
The average age of patients with identified NAr mutation included in this study was statistically significantly lower than that of patients in whom the mutation was not detected (p<0.05). No statistically significant differences were observed between patients with and without NAr mutations in terms of demographic information including gender, clinical laboratory data including AST, ALT, and AFP, liver histologies containing inflammation and fibrosis scores, and virological features including HBV DNA levels. (Tablo 2).
Discussion
This study aimed to investigate the prevalence, characteristics, and potential clinical implications of amino acid changes at the 42 positions associated with NAr within the HBV polymerase RT region in patients with CHB.
The 42 potential NAr mutations identified in the pre-treatment HBV RT region are classified into four categories: primary, secondary/compensatory, putative, and pretreatment mutations. Primary and secondary/compensatory resistance mutations are well-defined classical antiviral drug resistance mutations by in vivo and in vitro phenotypic experiments. Putative resistance mutations have their association with drug resistance defined, but these mutations have not yet been experimentally confirmed in vitro. In this regard, 26 putative RT mutation positions associated with NAr have been identified. Pretreatment mutations are as defined that the association between with drug resistance has not been yet evaluated [6, 7]. (Tablo 3).
In this study, mutations were identified in 35 different types at 21 out of the previously defined 42 NAr positions in the RT region of the HBV polymerase gene in 82 patients (80.39%). The prevalence of pre-treatment RT mutations reported in treatment-naive patients can vary considerably. These data indicate the presence of a high prevalence of NAr-associated mutations in the Pol/RT region, as previously reported, in treatment-naive patients with HBV/D genotypes. [7, 8].
The prevalence of primary and secondary drug resistance in the pre-treatment HBV RT region is reported to be less than 5%, with the most frequently observed mutations being rtM204I/V and rtL180M, respectively [9]. In this study, the prevalence of primary and secondary drug resistance was found to be 3.92% and 0.98%, respectively, in total 4.90%, which is less than 5%. However, the most frequently observed mutations included TDF-associated rtA194T and LAM-associated rtV173L, different from rtM204I/V and rtL180M, respectively (Tablo 3). These data indicate that spontaneous antiviral resistance is quite rare in treatment-naive patients with HBV/D genotypes included in this study, and there is no significant limitation to the use of NAs in patients with CHB.
In our study, a total of 26 mutations were identified in 15 out of the 26 putative positions associated with NAr in the HBV RT region, including 10 previously defined ones and 16 new mutations that differ from the previously defined ones.Out of the 6 pre-treatment RT mutation positions previously defined, treatment mutation was identified total 8 mutation in 5 of them. Additionally, 8 mutations were identified in total, including 7 new types that differ from the previously defined ones. The Y54H mutation was the most frequently observed variation. These data indicate a high prevalence of amino acid variations in the Pol/RT region of the HBV/D genotype. Further studies at the phenotypic level are needed, particularly to assess the relationship of the detected variations, including Y54H, with NAs.
The HBV RT region contains seven functional domains (G, F, A, B, C, D, and E) and five intermediate domains associated with these functional domains (F-A, A-B, B-C, C-D, and D-E) [6, 10, 11, 12 ]. Putative and pre-treatment mutations generally occur more frequently in the RT A-B intermediate domain, which overlaps with the ‘a’ determinant region of HBsAg. Thus, mutations occurring in the gene regions overlapping with the S gene in the RT region have the potential to alter HBV virulence, replication capacity, immune recognition, and antigenicity. Mutations in this domain, specifically at positions rt124, rt126, rt128, rt134, rt139, and rt153, have been reported as the most frequently detected variations [6, 7, 10, 11, 12]. In this study, the A-B interdomain of HBV RT was identified as the domain with the most frequent mutation variations. The rtD134N mutation, which overlaps with the HBsAg “a” determinant and can lead to the sI126N/S mutation, was identified in this domain. Additionally, mutations were detected at positions rt124, rt126, rt128, and rt139, which are reported as the most frequently observed variations in this domain.
In treatment-naive patients, 27 different polymorphic sites were identified in the HBV RT region. Among these, rt53, rt54, and rt91 are the most frequently genotype-dependent polymorphic amino acid sites across HBV A/B/C/D/E/F genotypes. Additionally, it has been reported that polymorphisms can be characterized by lower DNA levels [6, 8 ]. It has been indicated that the HBV/D genotype contains higher genetic diversity compared to other genotypes [13 ]. In this study, 13 out of the 21 amino acid positions with detected mutations were identified as genotype-associated polymorphic sites. And the previously described rt53, rt124, and rt256 were the most frequently polymorphic amino acid positions.
The effects of pre-treatment mutations occurring in the RT region in treatment-naive patients on the development and progression of progressive liver diseases such as HBV-related cirrhosis and hepatocellular carcinoma (HCC) have not yet been clearly defined. However, mutations such as rtL80I, rtD134N, rtN139K/T/H, rtY141F, rtM204I/V, rtF221Y, rtI224V, and rtM309K are suggested to be associated with the development of cirrhosis and HCC. [14-17]. In this study, mutations reported to be associated with progressive liver diseases such as cirrhosis and HCC, namely rtD134N in one patient, rtN139T in two patients, rtF221Y in eight patients, and rtM309K in eight patients, were detected. However, in this study, no statistically significant difference was observed between CHB patients with and without NAr mutations in terms of liver enzymes (AST, ALT, AFP) and liver damage-related data, including necroinflammatory activity and fibrosis stages (Tablo 2).
In some studies, the presence of pre-treatment Reverse Transcriptase (RT) mutations has been reported to be associated with lower Hepatitis B Virus (HBV) DNA levels in individuals with HBeAg-negative serostatus. Variations in the NAr-associated HBV RT region are suggested to impair viral replication by reducing polymerase enzyme activity [18, 19 ]. In this study, no statistically significant difference was observed in virological features, including HBV DNA levels, between CHB patients with detected and undetected NAr mutations. (Tablo 2).
The presence and prevalence of pre-treatment HBV RT mutations in CHB patients can be associated with epidemiological factors, HBV genotypes, viral replication dynamics, HBeAg serostatus, mutation detection methods, the presence of liver damage, and coexisting co-infections. However, the impact of pre-treatment HBV RT mutations on the progression of HBV-associated disease with antiviral treatment in CHB patients is not yet clear.
The data obtained from this study indicate a high prevalence of amino acid variations in the HBV RT region in patients with HBV/D genotype. However, it suggests limited effects on antiviral treatment and the progression of HBV-associated disease. Along with this also, the detection of mutations related to potential antiviral drug resistance in treatment-naive patients with chronic B hepatitis can contribute to the development of more effective treatment strategies and clinical management.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: None
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Trépo C, Chan HL, Lok A. Hepatitis B virus infection. Lancet. 2014;384(9959):2053-2063.
2. Sonderup MW, Spearman WC. Global disparities in hepatitis B elimination-a Focus on Africa. Viruses. 2022;14(1):82.
3. Summers J, Mason WS. Replication of the genome of a hepatitis B-like virus by reverse transcription of an RNA intermediate. Cell. 1982;29(2):403-15.
4. Lok AS, McMahon BJ, Brown RS Jr, Wong JB, Ahmed AT, Farah F. et al. Antiviral therapy for chronic hepatitis B viral infection in adults: a systematic review and meta-analysis. Hepatology. 2016;63(1):284-306.
5. Caligiuri P, Cerruti R, Icardi G, Bruzzone B. Overview of hepatitis B virus mutations and their implications in the management of infection. World J Gastroenterol. 2016;22(1):145-154.
6. Liu BM, Li T, Xu J, Li XG, Dong JP, Yan P, et al. Characterization of potential antiviral resistance mutations in hepatitis B virus reverse transcriptase sequences in treatment naïve Chinese patients. Antiviral Res. 2010;85(3):512-519.
7. Choi Y-M, Lee S-Y, Kim B-J. Naturally occurring hepatitis B virus reverse transcriptase mutations related to potential antiviral drug resistance and liver disease progression. World J Gastroenterol. 2018;24(16):1708-1724.
8. Mirandola S, Sebastiani G, Rossi C, Velo E, Erne EM, Vario A et al. Genotype-specific mutations in the polymerase gene of hepatitis B virüs potentially associated with resistance to oral antiviral therapy. Antiviral Research. 2012;96(3):422-429.
9. Zhang Q, Liao Y, Cai B, Li Y, Li L, Zhang J, et al. Incidence of natural resistance mutations in naïve chronic hepatitis B patients: a systematic review and meta-analysis. J Gastroenterol Hepatol. 2015;30(2):252-261.
10. Banerjee P, Chakraborty A, Mondal RK, Khatun M, Datta S, Das K. HBV quasispecies composition in Lamivudine-failed chronic hepatitis B patients and its influence on virological response to Tenofovir based rescue therapy. Sci Rep. 2017;17(7):44742.
11. Zheng J, Zeng Z, Zhang D, Yu Y, Wang F, Pan CQ. Prevalence and significance of Hepatitis B reverse transcriptase mutants in different disease stages of untreated patients. Liver Int. 2012;32(10):1535-1542.
12. Yamani LN, Yano Y, Utsumi T, Wasityastuti W, Rinonce HT, Widasari DI, et al. Profile of mutations in the reverse transcriptase and overlapping surface genes of hepatitis B virus (HBV) in treatment-naïve indonesian HBV carriers. Jpn J Infect Dis. 2017;70(6):647-655.
13. Chu CJ, Keeffe EB, Han SH, Perrillo RP, Min AD, Soldevila-Pico C, et al. Prevalence of HBV precore/core promoter variants in the United States. Hepatology 2003;38(3):619-628.
14. Kim JE, Lee SY, Kim H, Kim KJ, Choe WH, Kim BJ. Naturally occurring mutations in the reverse transcriptase region of hepatitis B virus polymerase from treatment-naïve Korean patients infected with genotype C2. World J Gastroenterol. 2017;23(23) 4222-4232.
15. Huang CJ, Wu CF, Lan CY, Sung FY, Lin CL, Liu CJ et al. Impact of genetic heterogeneity in polymerase of hepatitis B virus on dynamics of viral load and hepatitis B progression. PLoS One. 2013;8(7):e70169.
16. Li H, Jia J, Wang M, Wang H, Gu X, Fang M, et al. F221Y mutation in hepatitis B virus reverse transcriptase is associated with hepatocellular carcinoma prognosis following liver resection. Mol Med Rep. 2017;15(5):3292-3300.
17. Wu Y, Gan Y, Gao F, Zhao Z, Jin Y, Zhu Y et al. Novel natural mutations in the hepatitis B virus reverse transcriptase domain associated with hepatocellular carcinoma. PLoS One. 2014;9(5):e94864.
18. Zhu B, Wang T, Wei X, Zhuo Y, Liu A, Zhang G. Accumulation of mutations in reverse transcriptase of hepatitis B virus is associated with liver disease severity in treatment-naïve Chinese patients with chronic hepatitis B. Adv Clin Exp Med. 2017;26(7):1123-1129.
19. Kirishima T, Okanoue T, Daimon Y, Itoh Y, Nakamura H, Morita A. et al. Detection of YMDD mutant using a novel sensitive method in chronic liver disease type B patients before and during lamivudine treatment. J Hepatol. 2002;37(2):259-265.
Download attachments: 10.4328.ACAM.22047
Bulent Cakal, Bilger Cavus, Mehves Poda, Alp Atasoy, Mesut Bulakcı, Mine Gulluoglu, Filiz Akyuz. Potential antiviral drug resistance mutations in patients with treatment-naive chronic hepatitis B. Ann Clin Anal Med 2024;15(3):182-187
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
Histopathological examination of gunshot wounds
Ali Tavasli 1, Erdem Okdemir 2, Safa Celik 1, Muzaffer Yildirim 1, Muhammet Demir 1, Alper Keten 3, Mehmet Siddik Tuncay 4
1 Department of Forensic Medicine, Branch Office, Gaziantep, Turkey, 2 Department of Forensic Medicine, Gokyuzu OSGB Lmt., Gaziantep, Turkey, 3 Department of Forensic Medicine, Institute of Forensic and Traffic Medicine, Heidelberg University, Germany, 4 Department of Physical Medicine and Rehabilitation, Liv Hospital, Gaziantep, Turkey
DOI: 10.4328/ACAM.22049 Received: 2023-11-15 Accepted: 2023-12-18 Published Online: 2024-01-27 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):188-192
Corresponding Author: Ali Tavasli, Department of Forensic Medicine, Branch Office, Gaziantep, Turkey. E-mail: alitavasli20@gmail.com P: +90 533 957 20 35 Corresponding Author ORCID ID: https://orcid.org/0000-0002-7661-5357
This study was approved by the Ethics Committee of Council of Forensic Medicine, Istanbul (Date: 2015-01-27, No: 2015/14)
Aim: Deaths resulted from gunshot injuries have forensic characteristics. Autopsies are required in order to determine the number of shots, direction of shots, distance of shots, deadly wounds etc. Histopathologic examination on wounds is one of the methods that help investigations. In this study, we aimed to define histopathologic parameters of entrance-exit wounds in gunshot injuries. Moreover, we tried to determine usability of the defined parameters in entrance-exit wounds differentiation and estimation of shooting distance.
Material and Methods: 113 gunshot wounds of which 62 entrance and 51 exit wounds are investigated in our study. Necrosis on epidermis and dermis; presence of foreign body (black pigmented particles, gunpowder; textile fibers) on epidermis and dermis; bleeding on dermis and subcutaneous tissues; presence of fat, bone and muscle tissue on wound tract are evaluated histomorphologically.
Results: Skin necrosis, widespread dermal bleeding, textile fibers and gunpowder on wound tracts were statistically high on entrance wounds. Presence of fat, muscle and bone tissues on wound tracts and absence of skin necrosis, gunpowder particles and textile fibers were statistically high on exit wounds. It was observed that gunpowder particles and necrotic tissue between gunpowder decreases as shooting distance increases. It was found that attained data were aligned with the literature.
Discussion: Along with other methods, histopathologic assessments support the verification of diagnosis. Moreover, it is helpful in making decision on whether the wound is a gunshot injury or not; entrance-exit wound differentiation and estimation of shooting distance in cases of corpses without clothes and interfered corpses or wound characteristics could not be assessed macroscopically.
Keywords: Forensic Pathology, Gunshot Wound, Histology, Microscopy, Range Of Fire
Introduction
In gunshot wound cases, differentiating the entrance wounds from the exit wounds and estimating the range of fire are important for the reconstruction of crime scenes and determining the manner of death. Such evaluations are mainly performed by detecting the gunshot residues (GSR) macroscopically. In cases where there is no GSR, the range of fire is considered a distant shot. However, in cases involving medical interventions, decomposition, and burning, the detection of GSR becomes difficult [1, 2]. Various microscopic examination methods have therefore been conducted, and these have included hematoxylin and eosin (H&E) and sodium rhodizonate staining, and Fourier-transform infrared (FT-IR) microscopy [3, 4]. Dolinak et al pointed out that FT-IR microscopy could be of benefit in cases where macroscopic examination is difficult, such as medical intervention, decomposition, and burning [5]. Marty et al indicated that measuring the density and distribution of the staining reaction using sodium rhodizonate as a histological staining technique is an effective method to determine the range of fire and the entrance wound [6].
Perez and Molina’s study of H&E showed that a bullet can carry powder grains to the exit wound [7]. Microscopic examination of gunshot wounds is therefore not recommended for routine assessments and there is thus little data on this subject [8, 9]. For this reason, the inclusion of a large volume of relevant data will help guide the evaluation of selected cases. In this study, we discuss the findings obtained using a standard microscopic evaluation of gunshot wounds with a relatively large sample group. A total of 113 wounds from 45 cases were examined macroscopically and microscopically.
Material and Methods
This study was approved by the Ethics Committee of Council of Forensic Medicine, Istanbul (Date: 27/01/2015, No: 2015/14). All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. In this study, deaths determined to be the result of gunshot wounds by the Morgue Department of the Council of Forensic Medicine were evaluated prospectively. Wounds sustained via short-barrel guns were evaluated. The type of barrel used was determined based on the descriptions of the ammunition obtained from the autopsy and criminal laboratory reports. The time of death of the cases did not exceed 36 hours. A total of 113 wounds from 45 cases were examined macroscopically and microscopically. For entrance wounds in clothing areas, the range of fire was determined according to the findings of the crime scene and the criminal laboratory reports.
In cases where the bullet was not present in the body, descriptions of the bullet were obtained from the crime scene investigation reports. A scalpel was used to sample each lesion at a radius of about 3 cm and a depth of 2 cm from the center of the entrance or exit hole. The samples were fixed in 10% formaldehyde for at least 48 hours. Samples with a thickness of approximately 3 mm were taken from the fixed tissues in parallel with the bullet trajectory. After tissue tracking, the tissues were embedded in paraffin blocks. From the paraffin blocks, sections with a 3–5 μm thickness were taken using a microtome. The prepared samples were stained with H&E and evaluated under a light microscope.
For the entry and exit wounds, necrosis in the epidermis and dermis, foreign bodies in the epidermis and dermis (black-pigmented particle powder and textile fibers), bleeding in the dermis and subcutaneous tissue, and the presence of fat, muscle, and bone tissue in the wound tract were examined. In addition, the relationship between the range of fire and the diameter of the bullet was evaluated using the histomorphological parameters described above. The presence of necrosis in the epidermis was divided into two groups: mild and severe. If the partial necrosis of keratinocytes was limited in the wound edges, it was considered to be mild. Contrarily, if the necrosis was intense and covered large areas, it was evaluated as severe. Light eosinophilic changes in the collagen around the wound trauma in the dermis were considered mild dermal necrosis, and basophilic changes in the collagen were considered severe dermal necrosis. Severe necrotic connective tissue elements were evaluated as “compact” if they maintained integrity under the epidermis and were considered “fragmented” if they disintegrated in the wound path. Bleeding in the subcutaneous tissue and dermis were classified as “focal” if it was partial and “common” if it was broad.
Statistical Analysis
All the data were analyzed with SPSS version 14.0 (Statistical Package for the Social Sciences) software for Windows. Individual and aggregate data were summarized using descriptive statistics including mean, standart deviations and percentages. For initial evaluations and comparison of the data with normal distribution, the Kolmogorov-Smirnov test was used. A statistical analysis of the collected data was conducted via a chi-square test using SPSS version 21.0 (IBM Inc., NY/USA). The level of statistical significance was accepted as p<0.05.
Results
Among the 113 gunshot wounds, 62 (54.9%) were entrance wounds, and 51 (45.1%) were exit wounds. Of the 53 wounds for which the range of fire was evaluated by macroscopic examination, 18 (34%) were contact wounds, 9 (17%) were close-range wounds, and 26 (49%) were distant-range wounds. Of the 59 bullets that created entrance wounds, 1 (1.7%) was 6.35 mm in diameter, 23 (39%) were 7.65 mm in diameter, and 35 (59.3%) were 9 mm in diameter.
The relationship between epidermal/dermal necrosis and the entrance/exit wounds was examined (Figure 1).
While the incidence of severe epidermal/dermal necrosis in the entrance wounds was significantly higher than that in the exit wounds, the absence of epidermal/dermal necrosis in the exit wounds was significantly higher than that in the entrance wounds (p<0.001). The correlation of epidermal necrosis with the entrance and exit wounds is shown in Table 1. When the relationship between compact severe dermal necrosis and the entrance/exit wounds was examined (Figure 2), compact severe dermal necrosis was detected in 26 (41.9%) of the entrance wounds and in 2 (3.9%) of the exit wounds. The incidence of compact severe dermal necrosis in the entrance wounds was significantly higher than that in the exit wounds (p<0.001). Moreover, the incidence of powder grains in the epidermis and powder grains and textile fibers in the dermis of the entrance wounds was significantly higher than that in the exit wounds (p<0.001, Table 2). When the relationship between dermal hemorrhage and the entrance/exit wounds was examined, focal dermal hemorrhage was detected in 41 (66.1%) of the entrance wounds and 44 (86.3%) of the exit wounds. Diffuse dermal hemorrhage was detected in 21 (33.9%) of the entrance wounds and 7 (13.7%) of the exit wounds. The incidence of diffuse dermal hemorrhage in the entrance wounds was significantly higher than that in the exit wounds (p=0.014). When the relationship between the adipose tissue in the dermis and the entrance/exit wounds was examined, adipose tissue was detected in the dermis in 5 (8.1%) of the entrance wounds and 11 (21.6%) of the exit wounds. The incidence of adipose tissue in the dermis of the exit wounds was significantly higher than that in the entrance wounds (p=0.04). Furthermore, in 3 (4.8%) of the entrance wounds, stratified squamous epithelium fragments were detected in the dermis. When the relationship between the muscle and bone tissue in the dermis and the entrance/exit wounds was examined, muscle and bone tissues were detected in the dermis in 3 (4.8%) of the entrance wounds and 18 (35.3%) of the exit wounds. The incidence of muscle and bone tissues in the dermis of the exit wounds was significantly higher than that in the entrance wounds (p<0.001).
In terms of the range of fire, there was no significant difference between epidermal necrosis, dermal necrosis, dermal hemorrhage, and subcutaneous hemorrhage in the examination of the entrance wounds. For the entrance wounds, the incidence of powder grains in the epidermis of the contact wounds was significantly higher than that of the close- and distant-range wounds (p<0.001). The incidence of powder particles in the dermis of the contact and close-range wounds was also significantly higher than that of the distant-range wounds (p<0.001). Similarly, the incidence of necrotic tissue fragments surrounded by powder particles in the dermis of the contact and close-range wounds was significantly higher than that of the distant-range wounds (p<0.001). For the entrance wounds in clothed areas, the incidence of textile fibers in the dermis of the close- and distant-range wounds was significantly higher than that of the contact wounds (p=0.019). The microscopic findings according to the range of fire are shown in Table 3.
Discussion
This is the first study conducted by the Institute of Forensic Medicine on the use of histopathology in distinguishing entry and exit of gunshot wounds and determining the shooting distance. In the study, histopathological parameters that can be used in the distinction of entry-exit wounds and determination of shooting distance were defined. Differentiating gunshot entrance and exit wounds and the range of fire are important in the resolution of forensic cases. Although related studies in the literature are limited, histopathological evaluations have been shown to be useful in determining entry/exit wounds and the range of fire [9-12]. In their study, Perez and Molina showed that microscopic examinations were not suitable for routine evaluations because some false positives were evident [7].
Baptista et al reported that necrosis (burns) was seen in the epidermis in all the entrance wounds in their study, but not in all the exit wounds [13]. However, in our study, epidermal necrosis was detected in some exit wounds. We believe that this is due to the greater number of wounds examined and their diversity. According to our results, the presence of severe epidermal necrosis is indicative that a wound is an entrance wound, while the presence of mild or no epidermal necrosis signifies that a wound is an exit wound.
In the study of Perez and Molina, soot and powder grains were detected in 60% of the entrance wounds in the tissue sections and 21% of the exit wounds [7]. In another study by Baptista et al, dermal powder grains were detected in 21.5% of the entrance wounds but not in the exit wounds [13]. The presence of powder particles in the entrance wounds in our study was consistent with the data in the literature, yet powder grains were not found in the epidermis or dermis of the exit wounds. Although the number of cases in our study was relatively high, we surmise that the absence of powder grains in the exit wounds may be related to the nature of the ammunition used. The supply of ammunition in Turkey is carried out by a single government agency, and ammunition other than full metal jacket (FMJ) bullets is very rare. The possible carrying effect of hollow-point (HP) ammunition may have created the difference in the study of Perez and Molina. However, Hlavaty et al8 stated that there was no difference between HP and FMJ rounds.
In the study of Baptista et al, adipose and muscle and bone tissue were not seen in any of the entrance wounds although adipose tissue was seen in 50% and muscle and bone tissue in 42.8% of the exit wounds [13]. In our study, we found a higher rate of adipose, muscle and bone tissues in the dermis of the exit wounds, which is consistent with the literature, and we posit that the presence of adipose, muscle, and bone tissues in the entrance wounds may be an artifact appearance due to the flow of blood resulting from the position of the body.
Powder particles might have been detected by histological examination in entrance wounds but not macroscopically [14]. In the study by Perez et al, while gunpowder particles and soot were not observed macroscopically, 9 of the 27 entrance wounds revealed powder grains microscopically [7]. Similarly, in our study, 3 of the 26 entrance wounds showed no macroscopic evidence of powder grains, but powder grains were present in the dermis. It is possible that these grains may have been transported by the bullets since bullet wipe can be seen in entrance wounds regardless of the range of fire [10].
There is no evidence in the literature that the presence of necrosis in entrance wounds can be used to determine the range of fire. In our study, no statistically significant correlation was found between mild and severe epidermal/dermal necrosis rates and the range of fire. Additionally, no statistically significant correlation was found between the skin and subcutaneous hemorrhage rates and the range of fire.
Necrotic tissue fragments surrounded by black-pigmented particles were observed in 83.3% of the contact wounds, 77.8% of the close-range wounds, and 3.8% of the distant-range wounds, so we determined that the presence of GSR in the skin decreased as the range of fire increased. These findings are consistent with those in the literature.
In our study, the incidence of powder grains in the epidermis of the contact wounds was significantly higher than that in the epidermis of the close- and distant-range wounds, while their incidence in the dermis was significantly higher for the contact and close-range wounds than for the distant-range wounds. Hlavaty et al stated that there is no correlation between macroscopic and microscopic examinations in determining range of fire [15]. The most important factors determining the behavior of powder grains were not discerned in either study. The physical form of gunpowder grains affects the distance they travel, so the available data were insufficient to reach a scientific conclusion [10].
The use of standard histological examination methods in gunshot wound cases depends on the circumstances. Microscopic examinations are recommended in selected cases; however, there are diverse types of bullets and gunpowder. Such a large number of variables constitutes an obstacle to the establishment of diagnostic criteria for microscopic examination. Further studies in this regard are therefore warranted.
Conclusion
In conclusion, in cases of gunshot wounds, histopathological evaluation, in addition to other methods, is supportive in confirming the diagnosis. At the same time, we think that in cases where the integrity of the corpse is disrupted, clothing is absent, intervention has been carried out and/or wound characteristics cannot be evaluated macroscopically, histopathological examination is a method that should be used and will guide us in determining whether the wound is a gunshot wound or not, as well as in distinguishing between entry and exit wounds and estimating the distance.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: None
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Nishshanka MB, Paranirubasingam P, Shepherd C. A forensic-based study on low angled AK rifle bullet entry wounds using a porcine model. J Forensic Leg Med. 2020;74:102025.
2. Gentile G, Andreola S, Bailo P, Battistini A, Boracchi M, Tambuzzi S, et al. A brief review of scanning electron microscopy with energy-dispersive x-ray use in forensic medicine. Am J Forensic Med Pathol. 2020;41(4):280-6.
3. Gentile G, Tambuzzi S, Andreola S, Boracchi M, Gibelli L, Migliorini AS, et al. Is it possible to detect lead derived from gunshot residues on decalcified human bone by means of a histochemical staining with sodium rhodizonate? Forensic Sci Int. 2020;316:110474.
4. Pircher R, Grosse Perdekamp M, Mierdel K, Pollak S, Thierauf-Emberger A, Geisenberger D. Bullet wipe on the uppermost textile layer of gunshot entrance sites: may it be absent due to pre-existing blood staining? Int J Legal Med. 2019;133(5):1437-42.
5. Dolinak D, Wise SH, Jones C. Microscopic and spectroscopic features of gunpowder and its documentation in gunshot wounds in charred bodies. Am J Forensic Med Pathol. 2008;29(4):312-9.
6. Marty W, Sigrist T, Wyler D. Determination of firing distance using the rhodizonate staining technique. Int J Legal Med. 2002;116(1):1-4.
7. Perez DB, Molina DK. The utility of routine histological examination of gunshot wounds. Am J Forensic Med Pathol. 2012;33(3):231-3.
8. Giorgetti A, Giraudo C, Viero A, Bisceglia M, Lupi A, Fais P, et al. Radiological investigation of gunshot wounds: a systematic review of published evidence. Int J Legal Med. 2019;133(4):1149-58.
9. Baum GR, Baum JT, Hayward D, MacKay BJ. Gunshot wounds: Ballistics, pathology, and treatment recommendations, with a focus on retained bullets. Orthop Res Rev. 2022;14(9):293-317.
10. Euteneuer J, Gosch A, Cachee P, Courts C. Correction to: A distant relationship?-investigation of correlations between DNA isolated from backspatter traces recovered from firearms, wound profile characteristics, and shooting distance. Int J Legal Med. 2021;135(6): 2685.
11. de Martins de Souza AL, Carreteiro Damasceno J, Prado CBD, Magalhaes Gurgel MA, Carvalho Silva R. Uncertainty model for automated gunshot residue particle length measurements obtained from electron microscopy images. J Microsc. 2023;12(9):1-5.
12. Ledergerber TD, Feeney W, Arroyo L, Trejos T. A feasibility study of direct analysis in real time-mass spectrometry for screening organic gunshot residues from various substrates. Anal Methods. 2023;15(36):4744-57.
13. Baptista MV, d’Avila SC, d’Avila AM. Histopathological detection of entry and exit holes in human skin wounds caused by firearms. J Forensic Leg Med. 2014;25(7):49-52.
14. Petrus K, Angyal M, Toth D, Poor VS, Heckmann V, Simon G. Forensic assessment of a life-threatening penetrating abdominal air gun injury. Leg Med (Tokyo). 2023;60(9):1-4.
15. Hlavaty L, Roquero L, Amley J, Root K, Ishikawa M, Koopmeiners A, et al. Discordance of gross and histologic findings in estimating the range of fire of gunshot wounds. J Forensic Sci. 2019;64(5):1399-411.
Download attachments: 10.4328.ACAM.22049
Ali Tavasli, Erdem Okdemir, Safa Celik, Muzaffer Yildirim, Muhammet Demir, Alper Keten, Mehmet Siddik Tuncay. Histopathological examination of gunshot wounds. Ann Clin Anal Med 2024;15(3):188-192
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
The evolution of academic publications on pregnant women with COVID-19 and critical care: A holistic investigation of global outcomes with bibliometric analysis
Nevzat Mehmet Mutlu, Behiye Deniz Kosovalı
Department of Critical Care, University of Health Sciences, Ankara City Hospital, Ankara, Turkey
DOI: 10.4328/ACAM.22065 Received: 2023-12-06 Accepted: 2024-01-15 Published Online: 2024-01-22 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):193-198
Corresponding Author: Nevzat Mehmet Mutlu, Department of Critical Care, University of Health Sciences, Ankara City Hospital, 06800, Çankaya, Ankara, Turkey. E-mail: mutlunm@gmail.com P: +90 505 501 57 02 Corresponding Author ORCID ID: https://orcid.org/0000-0001-7981-3060
Aim: It was aimed to analyze scientific articles published on “pregnant with COVID-19, and critical care” by statistical and bibliometric methods.
Material and Methods: Articles published between 2019-2021 were obtained from the Web of Science database and analyzed using comprehensive bibliometric methods and citation analysis. Spearman correlation coefficient was used for correlation studies. Network visualization maps were created to analyze citations and identify trending topics.
Results: A total of 314 publications were found, of which 218 (69.4%) were articles. The top 5 contributors to the literature are the USA (32.5%), England (10.1%), Italy (8.7%), Spain (8.7%) and Turkey (7.7%). The top 3 journals with the most publications were BMJ Case Reports (4.1%), International Journal of Gynecology & Obstetrics (4.1%) and American Journal of Perinatology (3.7%).
Discussion: In this bibliometric study for “pregnant with COVID-19, and critical care”, it can be said that the trend topics are depression, stress, childbirth, ARDS, breastfeeding, newborn and maternal outcomes. This article can help scientists and clinicians evaluate new treatment and critical care strategies.
Keywords: Pregnancy, Pregnant, COVID-19, Critical Care, Bibliometric Analysis
Introduction
COVID-19 pneumonia was first reported in December 2019. It was later declared a global public health emergency by the World Health Organization [1,2]. At the beginning of the epidemic, the clinical features of pregnant with COVID-19 and non-pregnant adults were similar [1]. However, it has been reported that pregnant women with COVID-19 in the later period are more likely to be admitted to the critical care unit or need invasive mechanical ventilation compared to non-pregnant women of reproductive age. Ethnicity, chronic hypertension, diabetes, high maternal age and high body mass index were expressed as risk factors for severe COVID-19 pneumonia in pregnancy. It has also been reported that pregnant women diagnosed with COVID-19 are more likely to have premature birth and maternal death than pregnant women without COVID-19. Critical care follow-up may be required in cases of severe pneumonia, acute respiratory distress syndrome (ARDS), sepsis, septic shock, myocarditis, arrhythmia, cardiogenic shock and multi-organ failure in pregnant women with a diagnosis of COVID-19. Therefore, pregnant women with COVID-19 are considered to be a high-risk group [3,4,5]. Given that COVID-19 still persists and disease severity varies by variant, it is important to share experiences of pregnant women with COVID-19 and critical care.
Bibliometric research uncovers the most cited influential works, the most active authors, journals, and institutions in a topic or field. With keyword analyses obtained as a result of comprehensive statistical and bibliometric analyzes, researchers also give an idea about new studies that they can design by seeing past and current trends. Researchers who read the bibliometric outputs obtained as a result of the analysis of hundreds of studies published on a subject can dominate the literature in a short time [6-11].
We also thought that bibliometric analysis on pregnant with COVID-19, and intensive care would provide a better study of the impact of the disease on this population.
In this study, scientific articles published on “pregnant with COVID-19, and critical care” between 2019-2021 were analyzed using statistical and bibliometric methods. As a result of the analyses, it was aimed to determine the most cited influential studies, the most active authors, journals, institutions and countries, to reveal the cooperation between countries and trend topics, and to summarize the subject holistically.
Material and Methods
Web of Science (WoS) database (by Clarivate Analytics, USA) was used for literature review. The publication search was done in the Topic (title, abstract and keywords) section of the studies. Various keywords related to pregnancy, COVID-19 and critical care were used as search keywords in WoS. With this search method, in the title, abstract or in the keywords section, related to pregnancy, COVID-19 and critical care (pregnancy, pregnant, pregnancies, postpartum, post partum, maternal, COVID-19, coronavirus, SARS-CoV-2, n-CoV, critical care, critical care) were retrieved and downloaded from the WoS database. The search process was determined as 2019-2021 (access date: 01.11.2021). Due to the addition of new publications to the WoS database every day, the search findings may change at different access dates.
VOSviewer (Version 1.6.17, Leiden University’s Center for Science and Technology Studies) package program was used for bibliometric network visualizations [12]. The website (https://app.datawrapper.de) was used for the World map drawing, which was created to show the distribution of articles by world countries. Statistical analyzes were performed with the SPSS (Version 22.0, SPSS Inc., Chicago, IL, USA) package program. Whether the data were normally distributed or not was evaluated with the Kolmogorov-Smirnov test. In order to determine whether there is a relationship between the scientific work productivity of countries on pregnant with COVID-19, critical care and their economic power, the number of articles produced by the countries of the world and some economic development indicators of the world countries, gross domestic product (GDP), and GDP per capita (The World Bank-2020, Website accessed 10 October 2021, Available at: https://data.worldbank.org/indicator/NY.GDP.MKTP.CD,) were analyzed. Spearman correlation coefficient was used for the correlation analysis in accordance with the data distribution. Statistically significant difference limit was accepted as P<0.05.
Since this study is a bibliometric analysis, it does not require ethics committee approval.
Results
A total of 314 publications on “pregnant with COVID-19, and critical care” were found in the WoS database between 2019 and 2021. Of these publications, 218 (69.4%) were articles, 61 (19.4%) were reviews, 28 (8.9%) were letters, and the remainder were early access and editorial material. Bibliometric analyses were carried out with 218 articles from a total of 314 publications in the article category. Of these articles, 208 (95.4%) were published in English, 4 (1.8%) in Spanish, 3 (1.4%) in German, 2 (0.9%) in French, and 1 (0.5%) in Russian. The h-index of 218 articles was 20, average citations per article 9.2, sum of times cited 2005 (without self citations:1825).
1.1. Active Research Areas
Distribution of analyzed articles by research fields (at least 3 articles were published) was as follows: Obstetrics Gynecology (85, 38.9%), Medicine General Internal (52, 23.8%), Pediatrics (34, 15.5%), Infectious Diseases (14, 6.4%), Immunology (8, 3.6%), Multidisciplinary Sciences (7, 3.2%), Reproductive Biology (7, 3.2%), Health Care Sciences Services (6, 2.7%), Medicine Research Experimental (6, 2.7%), Public Environmental Occupational Health (6, 2.7%), Endocrinology Metabolism (5, 2.2%), Acoustics(4, 1.8%), Anesthesiology (4, 1.8%), Cardiac Cardiovascular Systems (4, 1.8%), Radiology Nuclear Medicine Medical Imaging (4, 1.8%) and Nursing (3, 1.3%).
1.2. Active Countries
The first 19 countries with the highest number of articles (publishing 5 or more articles) are as follows: USA (71, 32.5%), UK (23, 10.1%), Italy (19, 8.7%), Spain (19, 8.7%), Turkey (17, 7.7%), China (16, 7.3%), India (15, 6.8%), Brazil (10, 4.5%), Israel (10, 4.5%), Australia (8, 3.6%), Iran (8, 3.6%), Belgium (7, 3.2%), France (7, 3.2%), Germany (7, 3.2%), Chile (6, 2.7%), Mexico (6, 2.7%), Switzerland (6, 2.7) %), Argentina (5, 2.2%) and Sweden (5, 2.2%).
Cluster analysis was performed among 26 countries, which produced at least 3 articles from 60 countries that published articles on “pregnant with COVID-19, and critical care”, and whose authors have international cooperation, and it is shown in Figure-1.a. According to the results of the clustering analysis, 4 different clusters related to international cooperation were formed (Cluster-1:Austria, Belgium, Chile, Germany, Ireland, Israel, Italy, China, Spain, Switzerland, Turkey, USA. Cluster-2:Argentina, Brazil, Canada, France, India, Indonesia, Japan, Mexico, Pakistan. Cluster-3: Australia, England, Scotland. Cluster-4: Denmark, Sweden). In addition, the total link strength scores showing the cooperation power of 26 countries were calculated and the International collaboration density map created according to these scores is shown in Figure-1.b.
1.3. Correlation Analysis
There was a high level of statistically significant correlation between the number of articles produced by countries on “pregnant with COVID-19, and critical care” and GDP values, and a moderately positive correlation between GDP per capita values (respectively, r=0.702, p<0.001; r= 0.643, p<0.001).
1.4. Active Authors
The 3 most active authors who published 4 or more articles on that topic were Ferrazzi E. (5, 2.29%), Goffman D. (4, 1.83%), Tayman C. (4, 1.83%), respectively.
1.5. Active Institutions
The 7 most active institutions that have published 5 or more articles on the study topic were University of Milan (9, 4.1%), Tel Aviv University (8, 3.6%), Foundation IRCCS Ca’Granda Ospedale Maggiore Policlinico (Milan) (6, 2.7%), University of Health Science (Turkey) (6, 2.7%), Columbia University (5, 2.2%), King’s College London (5, 2.2%), and Weill Cornell Medicine (5, 2.2%).
1.6. Active Journals
218 articles published on study topic were published in 123 different journals. The top 20 most active journals producing 3 or more articles, the total number of citations received by the journals and the average number of citations per article are presented in Table-1.
1.7. Citation Analysis
Among the 218 articles published on the study topic, the first 15 articles with the highest number of citations, according to the total number of citations, are presented in Table 2.
1.8. Co-citation Analysis
All 218 articles analyzed had a total of 4123 cited studies in the references section. The first 4 studies that received the most co-citations (more than 30 citations) were Chen (2020) (Number of citation:NC=52), Knight (2020) (NC=39), Allotey (2020) (NC=34), and Di Mascio (2020) (NC=31) [1,3,5,13].
1.9. Trending topics
470 different keywords were used in all 218 articles published on the study topic. 54 different keywords that were used in at least 3 different articles are shown in Table-3. Trend network visualization map performed to reveal trend topics is shown in Figure-2 and citation network visualization map performed to reveal the most cited topics is shown in Figure-3.
Discussion
When the distribution of publications of the world countries on the productivity of articles on “pregnant with COVID-19, and critical care” is examined, the countries that published more than 10 active articles were the USA, England, Italy, Spain, Turkey, China, India, Brazil, Israel. Of these countries, 5 were developed and 4 (Turkey, China, India, Brazil) were developing or having large economies. In our study, a high correlation was found between article productivity and GDP, and a moderate relationship between GDP per capita. This shows that the economic size of the countries is effective in the efficiency of publications on “pregnant with COVID-19, and critical care”. In bibliometric studies conducted on many different medical subjects, it was concluded that economic power is effective in publication efficiency [6,8,10,12]. Considering the proportion of pregnant who need critical care during the pandemic, it is thought that the consequences of serious viral infection will probably be worse if the number of critical care units is insufficient. Therefore, the economic power of a country will be effective in both academic efficiency and pandemic management. When the density map created according to the total cooperation score between the countries is evaluated, it was determined that the countries with the most intensive cooperation were the USA, Spain, Italy, Germany, China, India, Switzerland, Brazil, Belgium, England, France and Ireland, respectively. When the co-authorship collaborations of countries were examined, it was seen that, contrary to the literature, geographical country-based collaborations did not have a significant effect on article production.
The journals that publish the most articles on the study topic are respectively BMJ Case Reports, International Journal of Gynecology & Obstetrics, American Journal of Perinatology, Cureus, Journal of Maternal-Fetal & Neonatal Medicine, American Journal of Obstetrics & Gynecology MFM, and Journal of Perinatal Medicine. We can recommend that authors who want to publish on this topic, first consider these journals. When the citation analyzes of the journals are evaluated, the most effective journals according to the average number of citations per article they publish are; Ultrasound in Obstetrics & Gynecology, Obstetrics and Gynecology, BJOG-An International Journal of Obstetrics and Gynaecology, Acta Obstetricia et Gynecologica Scandinavica, and the American Journal of Obstetrics and Gynecology. We can recommend that researchers who want their articles to be cited more should primarily consider these journals.
The articles were evaluated according to their total and annual average number of citations. The most influential first study was published in BMJ by Knight et al. [3]. The other most influential studies on this topic was done by Zimmermann and Curtis, Ferrazzi et al., Grohskopf et al. and Collin et al. [14,15,16,17]. Apart from these studies, when the articles are evaluated according to the average number of publications per year, Saccone et al. and Villar et al. were prominent[18,19]. According to the co-citation numbers of all analyzed articles, the studies of Chen, Knight, Allotey, and Di Mascio were determined as the most effective studies [1,3,5,13]. We recommend that clinicians and researchers interested in this subject read these publications first.
As a result of the evaluation of keyword analysis and clustering analysis, it was seen that our study subject was clustered in 6 different colors. The most cited keywords were transmission, symptoms, epidemiology, morbidity, obesity and delivery. According to the results of the analysis made to determine the trend topics, it was determined that the keywords studied in the last months were depression, stress, birth, ARDS, breastfeeding, neonatal and maternal outcomes.
When we evaluated trend issues within the scope of the literature, Chen et al. stated that none of the nine pregnant women in the third trimester included in the study had intrauterine fetal infection due to vertical transmission of COVID-19 infection [1]. In the United Kingdom, it was reported that one out of every 10 pregnant women admitted to the hospital due to SARS-CoV-2 infection needed respiratory support in the critical care unit and 1% of the cases were mortal. Most of the admitted pregnant women were in the second or third trimester, and the transmission rate from pregnant to baby was 5% [3]. A meta-analysis conducted in 2020 revealed that there was no vertical transmission from pregnant to their babies, and that the most common complication in pregnant women was preterm delivery, and the rates of preeclampsia, cesarean section and perinatal death were also increased [13]. In a study conducted in New York in 2021, it was reported that there was no mother-to-baby transmission by being in the same room and breastfeeding [20]. There are studies supporting that SARS-CoV-2 has not been detected in breast milk. As a result of these studies, it is recommended that mothers with COVID-19 continue to breastfeed [1,20,21].
As for outcomes, another trending topic, we can say that maternal deaths, stillbirths, ruptured ectopic pregnancies and maternal depression due to the COVID-19 pandemic have increased, according to the results of a systemic review and meta-analysis conducted in 2021. In this study, the observed increase in maternal mortality was based on data from low-income and middle-income countries. According to the rapid report ‘Mothers and Babies: Reducing Risk through Audits and Confidential Inquiries across the UK,’ maternal deaths associated with SARS-CoV-2 (9.9 per 100,000) in the first wave of the pandemic (March-May 2020) compared to pre-pandemic (2016-2018)(9.7 per 100,000) was found to be higher [22,23]. In another study evaluating the deaths of mothers with COVID-19, it was found that the risk of mortality was 2 times (RR: 2.26, 95% CI: 1.77-2.89) in pregnant without comorbidity, and 5 times (RR: 5.09, 95% CI: 2.00-12.98) in pregnant with at least one comorbidity (especially the presence of diabetes and obesity) has been reported to increase [24]. In a study conducted in Mexico, the mortality rate in hospitalized pregnant with COVID-19 was found to be 2.8%, and it was stated that obesity and diabetes increase mortality. However, in this study, unlike the others, it was stated that not all maternal deaths were directly related to COVID-19, and limited health services also contributed to mortality [25]. There are few publications on “pregnant with COVID-19, and critical care” in low and middle-income countries, and it is necessary to support these countries in research in order to create strategies in this regard.
Strengths and limitations of the study
The fact that we used only the WoS database in the literature review in our study is a limitation. Reasons for using only the WoS database are that citation and co-citation analysis that can be done in the WoS database cannot be done in the Pubmed database. The reason why the Schopus database is not used is because journals with lower impact levels are indexed compared to the WoS database.
Conclusion
We shared a summary of 218 articles published between 2019-2021 in this comprehensive bibliometric study on “pregnant with COVID-19, and critical care”. It was determined that the trend topics were depression, stress, birth, ARDS, breastfeeding, neonatal outcome and maternal outcomes. By presenting a summary of the global outputs on “pregnant with COVID-19, and critical care”, this article can be a useful resource for scientists and clinicians to evaluate treatments and critical care strategies for global viral infections such as SARS-CoV-2 infections.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: None
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Chen H, Guo J, Wang C, Luo F, Yu X, Zhang W, et al. Clinical characteristics and intrauterine vertical transmission potential of COVID-19 infection in nine pregnant women: A retrospective review of medical records. The lancet, 2020;395(10226):809-815.
2. Wang LM , Lai SP, Liang, SJ, Yang, ST, Liu, CH, Wang PH. Maternal and fetal outcomes of the pregnant woman with COVID-19: The first case report in Taiwan. Taiwan J Obstet Gynecol, 2021;60(5):942-944.
3. Knight M, Bunch K, Vousden N, Morris E, Simpson N, Gale C, et al. Characteristics and outcomes of pregnant women admitted to hospital with confirmed SARS-CoV-2 infection in UK: national population based cohort study. BMJ. 2020;369:2107.
4. Favre G, Pomar L, Musso D, & Baud D. 2019-nCoV epidemic: What about pregnancies?. Lancet (London, England), 2020;395(10224):40.
5. Allotey J, Stallings E, Bonet M, Yap M, Chatterjee S, Kew T, et al. Clinical manifestations, risk factors, and maternal and perinatal outcomes of coronavirus disease 2019 in pregnancy: Living systematic review and meta-analysis. BMJ. 2020;370:3320.
6. Kosovali BH, Mutlu NM. Global scientific outputs of extracorporeal membrane oxygenation in COVID-19: A bibliometric overview. Perfusion. 2023;38(6):1153-1164.
7. Mutlu-Sagesen HLE, Sagesen EA, Ozcan M. Bibliometric analysis of zirconia publications between 1980 and 2021: Global productivity and publication trends. J Prosthodont Res. 2024;68(1):147-155.
8. Erol ME, Özyalçın S. Global scientific outputs of tricuspid valve publications: A bibliometric approach. Turkish Journal of Clinics and Laboratory. 2021;12(3):288-296.
9. Kiraz M, Demir E, Özdemir Ö. An international bibliometric study of scientific articles on intracranial aneurysms. The Neuroradiology Journal. 2021;34(5):482-493.
10. Mutlu-Sagesen HLE, Sagesen A. The evolution of esthetic publications in dentistry, research trends and global productivity: A bibliometric analysis. Int J Prosthodont. Epub 2023 May 30.
11. Golpinar M, Demir E. Global research output of the cerebellum: Yesterday, today, and tomorrow. Journal of the Anatomical Society of India. 2020;69(3):155-165.
12. Van Eck NJ. Waltman L. Software survey: VOSviewer, a computer program for bibliometric mapping. Scientometrics. 2010;84(2):523–538.
13. Di Mascio D, Khalil A, Saccone G, Rizzo G, Buca D, Liberati M, et al. Outcome of coronavirus spectrum infections (SARS, MERS, COVID-19) during pregnancy: A systematic review and meta-analysis. Am J Obstet Gynecol MFM. 2020;2(2):100-107.
14. Zimmermann P & Curtis N. COVID-19 in children, pregnancy and neonates: A review of epidemiologic and clinical features. The Pediatric Infectious Disease Journal. 2020;39(6):469.
15. Ferrazzi E, Frigerio L, Savasi V, Vergani P, Prefumo F, Barresi S, et al. Vaginal delivery in SARS-CoV-2-infected pregnant women in Northern Italy: A retrospective analysis. BJOG: An International Journal of Obstetrics & Gynaecology. 2020;127(9):1116-21.
16. Grohskopf LA, Alyanak E, Broder KR, Blanton LH, Fry AM, Jernigan DB & Atmar RL. Prevention and control of seasonal ınfluenza with vaccines: Recommendations of the advisory committee on ımmunization practices – United States, 2020-21 Influenza Season. MMWR. Recommendations and reports: Morbidity and mortality weekly report. Recommendations and reports. 2020;69(8):1–24.
17. Collin J, Byström E, Carnahan A, & Ahrne M. Public Health Agency of Sweden’s Brief Report: Pregnant and postpartum women with severe acute respiratory syndrome coronavirus 2 infection in critical care in Sweden. Acta obstetricia et gynecologica Scandinavica. 2020;99(7):819-822.
18. WAPM (World Association of Perinatal Medicine) Working Group on COVID-19. Maternal and perinatal outcomes of pregnant women with SARS-CoV-2 infection. Ultrasound Obstet Gynecol. 2021;57(2):232-241. Erratum in: Ultrasound Obstet Gynecol. 2021;58(3):496.
19. Villar J, Ariff S, Gunier RB, Thiruvengadam R, Rauch S, Kholin A, Papageorghiou AT. Maternal and neonatal morbidity and mortality among pregnant women with and without COVID-19 infection: the INTERCOVID multinational cohort study. JAMA Pediatrics. 2021;175(8):817-826.
20. Dumitriu D, Emeruwa UN, Hanft E, Liao GV, Ludwig E, Walzer L, et al. Outcomes of neonates born to mothers with severe acute respiratory syndrome Coronavirus 2 infection at a large medical center in New York City. JAMA Pediatrics. 2021;175(2):157–167.
21. Wu Y, Liu C, Dong L, Zhang C, Chen Y, Liu J, et al. Coronavirus disease 2019 among pregnant Chinese women: case series data on the safety of vaginal birth and breastfeeding. BJOG: An International Journal of Obstetrics & Gynaecology. 2020;127(9):1109-1115.
22. Chmielewska B, Barratt I, Townsend R, Kalafat E, van der Meulen J, Gurol-Urganci I, et al. Effects of the COVID-19 pandemic on maternal and perinatal outcomes: A systematic review and meta-analysis. The Lancet Global Health. 2021;9(6):759-772.
23. Knight M, Bunch K, Cairns A, Cantwell R, Cox P, Kenyon S, et al. Saving lives, improving mothers’ care. Rapid report: learning from SARS-CoV-2-related and associated maternal deaths in the UK. March–May, 2020. Oxford: National Perinatal Epidemiology Unit, University of Oxford 2020.
24. La Verde M, Riemma G, Torella M, Cianci S, Savoia F, Licciardi F, et al. Maternal death related to COVID-19: A systematic review and meta-analysis focused on maternal co-morbidities and clinical characteristics. International Journal of Gynecology & Obstetrics. 2021;154(2):212-219.
25. Mendez-Dominguez N, Santos-Zaldívar K, Gomez-Carro S, Datta-Banik S & Carrillo G. Maternal mortality during the COVID-19 pandemic in Mexico: A preliminary analysis during the first year. BMC Public Health. 2021;21(1):1-9.
Download attachments: 10.4328.ACAM.22065
Nevzat Mehmet Mutlu, Behiye Deniz Kosovalı. The evolution of academic publications on pregnant women with COVID-19 and critical care: A holistic investigation of global outcomes with bibliometric analysis. Ann Clin Anal Med 2024;15(3):193-198
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
Effect of moderate exercise and vitamin D3 on adipose tissue SIRT1 Protein in obese rats
Hanan Salih Abdulrraziq 1, Hala Mahmoud Ibrahim Abou Heif 2, Hala Mohamed El-Sayed Maklad 2, Gehan Yassin Soliman Shoeib 2, Mona Hassan Fathelbab 3
1 Department of Medical Physiology, Faculty of Medicine, Omar AlMukhtar University, Albayda, Libya, 2 Department of Medical Physiology, Faculty of Medicine, Alexandria University, Alexandria, Egypt, 3 Department of Medical Biochemistry, Faculty of Medicine, Alexandria University, Alexandria, Egypt
DOI: 10.4328/ACAM.22073 Received: 2023-12-16 Accepted: 2024-01-22 Published Online: 2024-01-26 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):199-203
Corresponding Author: Hanan Salih Abdulrraziq, Department of Medical Physiology, Faculty of Medicine, Omar AlMukhtar University, Albayda, Libya. E-mail: hnansaleh83@gmail.com P: +20 101 671 66 43 Corresponding Author ORCID ID: https://orcid.org/0009-0006-2771-6545
This study was approved by the Ethics Committee of Alexandria University (Date: 2022-06-16, No: 0201690)
Aims: Obesity is a chronic metabolic disease affecting almost all physiological functions of the body leading to increased morbidity and mortality. The increased prevalence of obesity has been attributed to several risk factors that contribute to several metabolic dysfunctions.
This study was conducted to evaluate the effect of moderate exercise and vitamin D3 on adipose tissue SIRT1 protein level in obese rats.
Material and Methods: The study was carried out on 40 adult male Wister albino rats, divided into Group 1 (control group): 8 rats received a standard rat diet for 12 weeks and Group 2 (obese group): 32 rats received a high fat diet (HFD) for 4 weeks. Group 2 was further subdivided into 4 subgroups each of 8 rats; (a) Control sedentary group (b) Exercise group (c) Vitamin D3 supplement group (d) Exercise plus vitamin D3 supplement group. At the end of 12 weeks, the anthropometric index and adipose tissue SIRT1 protein by ELISA were measured.
Results: HFD results in abnormal anthropometric data and a significant reduction of SIRT1 protein in adipose tissue. In addition to elevation of SIRT1 protein in combined treatment, there was a negative correlation between SIRT1 protein and adiposity index.
Discussion: The combination of VD and exercise improved significantly anthropometric parameters and increased adipose tissue SIRT1 protein.
Keywords: Obesity, VD3, Exercise, SIRT1
Introduction
Obesity is the outcome of complex relations between environmental, genetic, and epigenetic factors. The two main factors that are thought to contribute to obesity are excessive consumption of high-energy foods and physical inactivity [1].
One potential tactic for the prevention and/or treatment of obesity and its related complications has been to focus on the development and functional alterations of adipose tissues during the obesity process. Obese people frequently have low vitamin D levels, which are associated with a higher risk of obesity [2]. Sirtuins (SIRT1) is a family of NAD-dependent enzymes that deacetylate different proteins’ lysine residues. It seems that adipose tissue is a significant SIRT1 action site. It has been documented that a high-fat diet (HFD) increased insulin resistance by causing the proteolytic cleavage of SIRT1 [3].
Exercise is a crucial non-pharmacological tool that has a profoundly positive impact on the body’s many functional systems. Nevertheless, the precise process by which regular exercise improves organ function is still unclear.
As a result, this research was performed to explore their effect on SIRT1 protein in adipose tissue in a rat model of HFD-induced obesity.
Material and Methods
2.1 Experimental design:
The present study involved 40 healthy male Wister albino rats aged 2-3 months with body weights from 150- 180 g each. Animals were housed in separate cages. They were maintained in normal light and temperature conditions with unrestricted access to food and water. Before the study began, the animals were allowed to adjust to their new housing for a week. All institutional and national guidelines for the care and use of laboratory animals were followed.
All the animals were randomly divided into two groups. Group 1, the normal control group (NC), included 8 rats. They served as healthy controls and received a standard rat diet for 12 weeks. Group 2, the high-fat diet group (HFD), included 32 rats and received HFD for 4 weeks. The HFD was used to make rats obese, and it contained 20 g of fat per 100 g of food (To supply essential fatty acids, use 1 g of soybean oil and 19 g of butter oil) [4].
Then Group 2 was subdivided into 4 subgroups; each of 8 rats. Group 2A: sedentary HFD control group (Sed): rats received HFD for another 8 weeks; Group 2B: Exercise group (Exe): rats received HFD and did swimming exercise for 8 weeks; Group2 C: Vitamin D supplement group (VD): rats received HFD and were given oral VD supplementation (10 mcg/kg /day) [5] for 8 weeks purchased from (medical union pharmaceutical, Egypt). In addition, Group 2 D: Exercise plus vitamin D supplement group (combined): rats received HFD, vitamin D (10 mcg/kg /day) for 8 weeks and did swimming exercise for 8 weeks.
2.2 Swimming exercise protocol.
In the exercised group, rats were subjected to free swimming in a pool filled with 32 ± 1°C water. [6] The protocol was conducted 5 days/ week between 9 and 12 a.m. for 8 weeks. Each swimming session lasted for 30 minutes [6].
2.3 Anthropometric measurement
The rats’ weights were measured weekly from the beginning of the experiment. At the end of the experiment body length, weight, body mass index( BMI) and Lee index= cube root of final body weight (g)/ length (cm) [7] were measured for all groups. Furthermore, after anesthesia (isoflurane 4% inhalation) visceral white adipose tissues from the epididymal, mesenteric, perirenal, and retroperitoneal regions were dissected, weighed and the adiposity index was calculated (total visceral fat/final body weight) × 100) [8].
2.4 Determination of SIRT1 protein in adipose tissue.
A. Adipose tissue preparation
After removing any remaining blood, visceral adipose tissue was stored at -80 °C after being cleaned with pre-cooling PBS buffer (0.01M, pH=7.4). After homogenizing the frozen tissues in 100 mg tissue/ml cold PBS with a protease inhibitor cocktail (Sigma-Aldrich, St. Louis, MO. USA), the samples were centrifuged for 15 minutes at 12000 xg, and aliquots of the supernatant were taken for ELISA analysis to determine the presence of SIRT1 protein [9].
B. Determination of adipose tissue SIRT1 protein by ELISA.
Adipose tissue supernatant was used to determine adipose tissue SIRT1 protein using Rat SIRT1 ELISA Kit (Catalog No. ER1338, Fine Test, Wuhan, China, e-mail: www.fn-test.com) based on sandwich enzyme-linked immune-sorbent assay technology according to the manufacture instructions. The samples were analyzed in duplicate and SIRT1 protein was expressed in ng/ml. In addition the supernatant of adipose tissue homogenate was used for the estimation of total protein by Lowry’s method using folin phenol reagent with bovine serum albumin as a standard and the samples were analyzed in duplicate [10].
The results of adipose tissue SIRT1 ELISA was normalized to total tissue proteins and expressed in ng\mg protein.
Statistical analysis of the data
Data were analyzed using IBM SPSS software package version 20.0. (Armonk, NY: IBM Corp). To confirm that the distribution was normal, the Shapiro-Wilk test was performed. Measures like mean and standard deviation were used to describe quantitative data. The results were deemed significant at the 5% level. The Post Hoc test (Tukey), one-way ANOVA test, and Pearson coefficient were utilized.
Ethical approval
This study was approved by Ethics committee of the Alexandria University (Date: 2022-06-16, No: 0201690).
Results
3.1 Anthropometric parameters
A. Comparison between the different studied groups according to body weight in (gm) in week 1, 4 and 12: In week 1 no significant difference between studied groups (P = 0.885) . while In week 4 There was a significant increase in body weight in groups that took HFD in comparison to normal control group (P <0.001). In addition, in week 12 exercise and combined groups showed no significant difference when compared to normal control groups and a significant decrease in body weight when compared to the sedentary HFD group.
B- Comparison between the different studied groups according to (Adipose tissue weight, BMI, lee index and adiposity index): The mean of adipose tissue weight, BMI, lee index and adiposity index showed a significant difference between studied groups.in addition exercise and combined groups showed no significant difference when compared to normal control groups and a significant decrease in adipose tissue weight, BMI, lee index and adiposity index when compared to sedentary HFD group. (table1)
3.2 Adipose tissue SIRT1 protein.
There was a significant decrease in SIRT1 protein in the sedentary HFD group in comparison to the normal control group, while no significant difference in exercise and VD groups in comparison to normal control group (table 2).
In addition, there was a significant increase in SIRT1 protein in combined group in comparison to normal control group.
Furthermore, there was a significant increase in SIRT1 protein in VD, exercise and combined group in comparison to sedentary HFD group and its level showed no significant difference between VD and exercise groups.
Our study showed that SIRT1 protein had a significant negative correlation with Adiposity index (r=-0.415, p=0.008) in total sample (n =40)(figure1)
Discussion
Animal obesity brought on by a high-fat diet has been documented in a number of studies, and it is regarded as the most widely used and trustworthy model for researching obesity.
In the present study, as expected, a significant increase in anthropometric indices was observed in sedentary HFD control compared to normal control (NC) rats.
In support to our study Madkhali et al proved that, when compared to rats fed a standard diet (SD), rats fed an HFD for 12 weeks had significantly higher body weights, BMIs, Lee indices, and food intake [11]. Furthermore, Yang et al found that the body weight, adiposity index were significantly increased in HFD group [12].
Contradictory to our result Roza et al discovered that, in contrast to age-matched normal diet (ND) rats, the female HFD-treated animals displayed a decrease in body weight during the follow-up (8-week) period, even with higher calorie intake, after the fifth week of life. This result could be due to difference in period of study and animal sex [13].
In the current study moderate swimming exercise was effective in reducing anthropometric indices as compared to the sedentary HFD control group.
Exercise interventions are beneficial for overweight or obese people in terms of lowering body weight, BMI, and body fat percentage, according to a previous meta-analysis of three studies [14]. Lim et al found that anthropometric indices including weight, body fat mass (BFM), BMI, and weight circumference (WC) significantly decreased in older women following a 12-week period of aquatic exercise [15].
In the current study, VD supplementation decreased the anthropometric indices as compared to sedentary HFD control group.
Consistent with our study Verma et al found that VD decreases body weight, various adipose tissues weight, BMI, Lee index, as compared to HFD-treated rats [5].
Sergeev verified that a decrease in the weight of white adipose tissue is correlated with an increase in VD3 intake in a mouse model of induced obesity [16].
A contradictory study by Salehpour et al on 85 women with BMI ≥ 25 kg/m2 showed that VD supplementation (VD3 supplement tablet of 25 μg/d)for 12 weeks did not affect body weight, BMI, and percentage body fat (PBF), this could be due to low dose of VD used in the study [17].
In the current study, the combined group showed a significant reduction in anthropometric indices when compared to the sedentary HFD control group.
Kim et al showed that VD supplementation and exercise attenuated body weight gain, abdominal adiposity, and non-alcohol fatty liver (NAFLD) in rats following induction of obesity [18].
Salarinia et al, demonstrated that in women with type 2 diabetes, exercise and VD supplementation had positive synergistic effects on the cardiometabolic profile. The 12-week intervention with exercise plus VD resulted in significant reductions in body weight, BMI and body fat mass [19].
In the present study, HFD caused a significant reduction in adipose tissue SIRT1 protein level as compared to NC group. In addition SIRT1 showed significant negative correlation with adiposity index.
Consistent to our study, Wei et al discovered that the SIRT1 gene and protein expression are decreased in white adipose tissue (WAT) and brown adipose tissue (BAT) upon the administration of a high-fat non-ketotic diet [20].
Another study investigated whether HFD affects the levels of SIRT1. The results of the quantitative RT-PCR analysis of RNA extracted from WAT revealed that mice given an HFD had two times lower levels of SIRT1 mRNA [3].
In the current study, exercise and VD showed a significant elevation in SIRT1 protein level in AT as compared to sedentary HFD control group.
Liu et al demonstrated that moderate aerobic exercise had positive effects on the liver and kidney functions of diabetic mice. Elevated acetylation of nuclear factor κ (NF-κB) is linked to decreased SIRT1 expression, while SIRT1 expression in the kidney and liver is restored during exercise, which suppresses NF-κB activity [21].
In addition, Vargas et al stated that physical activity can boost oxidative metabolism, mitochondrial biogenesis, and SIRT1 expression in skeletal muscles [22].
VD up-regulate SIRT1 protein expression through up-regulation of adiponectin and consequent activation of activated mitogen phosphokinase (AMPK) [23].
Safarpour et al showed that VD supplementation for 8 weeks increases the serum level of SIRT1 Compared to placebo [24].
In addition, Chang et al suggested that VD supplementation may help to improve the increased intramyocellular fat deposition and related muscle mitochondrial changes that come with obesity, while also increasing the activity of AMPK and SIRT1 [25].
Conclusion
HFD induced obesity in rats resulted in abnormal anthropometric parameters. Furthermore, HFD induced a significant decrease in SIRT1 protein in adipose tissue compared to normal rats. In addition, treatment of HFD by exercise or combined treatment (VD+ exercise) normalized the anthropometric abnormalities in the treated groups versus sedentary HFD. This treatment also increases SIRT1 protein in adipose tissue of rats.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: None
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Pereira-Lancha LO, Campos-Ferraz PL, Lancha AH. Obesity: considerations about etiology, metabolism, and the use of experimental models. Diabetes Metab Syndr Obes. 2012;5:75-87.
2. Vranić L, Mikolašević I, Milić S. Vitamin D deficiency: consequence or cause of obesity? Medicina. 2019;55(9):541.
3. Chalkiadaki A, Guarente L. High-fat diet triggers inflammation-induced cleavage of SIRT1 in adipose tissue to promote metabolic dysfunction. Cell Metab. 2012;16(2):180-8.
4. Zhang XY, Guo CC, Yu YX, Xie L, Chang CQ. Establishment of high-fat diet-induced obesity and insulin resistance model in rats. Beijing Da Xue Xue Bao Yi Xue Ban. 2020;52(3):557-63.
5. Verma A, Goyal A, Kaur R, Kamboj A, Jain UK. Beneficial effect of vitamin d on high-fat diet-induced obesity in wistar rats. Asian J Pharm Clin Res. 2016;3:337-40.
6. Tunca U, Saygin M, Ozmen O, Aslankoc R, Yalcin A. The impact of moderate-intensity swimming exercise on learning and memory in aged rats: The role of Sirtuin-1. Iran J Basic Med Sci. 2021;24(10):1413-20.
7. Novelli EL, Diniz YS, Galhardi CM, Ebaid GM, Rodrigues HG, Mani F, et al. Anthropometrical parameters and markers of obesity in rats. Lab Anim. 2007;41(1):111-9.
8. Leopoldo AS, Lima-Leopoldo AP, Nascimento AF, Luvizotto RA, Sugizaki MM, Campos DH, et al. Classification of different degrees of adiposity in sedentary rats. Braz J Med Biol Res. 2016;49(4):e5028.
9. Zahedi H, Hedayati M. The effect of 14 weeks aerobic exercise on resveratrol supplementation on protein UCP-1, SIRT1, PGC-1α in liver tissue, subcutaneous and visceral fat tissue in male wistar rats. J Sport Biosci. 2018;10(1):39-58.
10. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193(1):265-75.
11. Madkhali HA. Morin attenuates high-fat diet induced-obesity related vascular endothelial dysfunction in Wistar albino rats. Saudi Pharm J. 2020;28(3):300-7.
12. Yang H, Xie J, Wang N, Zhou Q, Lu Y, Qu Z, et al. Effects of Miao sour soup on hyperlipidemia in high-fat diet-induced obese rats via the AMPK signaling pathway. Food Sci Nutr. 2021;9(8):4266-77.
13. Roza NA, Possignolo LF, Palanch AC, Gontijo JA. Effect of long-term high-fat diet intake on peripheral insulin sensibility, blood pressure, and renal function in female rats. Food Nutr Res. 2016;60:28536.
14. Stoner L, Rowlands D, Morrison A, Credeur D, Hamlin M, Gaffney K, et al. Efficacy of exercise intervention for weight loss in overweight and obese adolescents: meta-analysis and implications. Sports Med. 2016;46(11):1737-51.
15. Lim BO, Kang SS, Cho JH, Moon JW. The effect of combined aquatic exercise on body composition, muscular function, static balance and visual analogue scale in female elderly with knee osteoarthritis. Asian J Kinesiol. 2018;20(2):1-13.
16. Sergeev IN. Vitamin D-Cellular Ca(2+) link to obesity and diabetes. J Steroid Biochem Mol Biol. 2016;164:326-30.
17. Salehpour A, Hosseinpanah F, Shidfar F, Vafa M, Razaghi M, Dehghani S, et al. A 12-week double-blind randomized clinical trial of vitamin D₃ supplementation on body fat mass in healthy overweight and obese women. Nutr J. 2012;11:78.
18. Kolieb E, Maher SA, Shalaby MN, Alsuhaibani AM, Alharthi A, Hassan WA, et al. Vitamin D and swimming exercise prevent obesity in rats under a high-fat diet via targeting FATP4 and TLR4 in the liver and adipose tissue. Int J Environ Res Public Health. 2022;19(21):13740.
19. Salarinia M, Azizi M, Tahmasebi W, Khalvandi H. Effect of eight weeks of vitamin D supplementation and water-based exercise on cardiometabolic profile in women with type 2 diabetes. Sci Sport. 2023;38(3):283-92.
20. Wei X, Jia R, Wang G, Hong S, Song L, Sun B, et al. Depot-specific regulation of NAD(+)/SIRTs metabolism identified in adipose tissue of mice in response to high-fat diet feeding or calorie restriction. J Nutr Biochem. 2020;80:108377.
21. Liu HW, Kao HH, Wu CH. Exercise training upregulates SIRT1 to attenuate inflammation and metabolic dysfunction in kidney and liver of diabetic db/db mice. Nutr Metab (Lond). 2019;16:22.
22. Vargas-Ortiz K, Pérez-Vázquez V, Macías-Cervantes MH. Exercise and Sirtuins: A way to mitochondrial health in skeletal muscle. Int J Mol Sci. 2019;20(11):2717.
23. Nemeth Z, Kiss E, Takacs I. A healthy balance of homeostasis by epigenetic regulator SIRT1 may prevent the development of a specific” soil” that supports metabolic disorders and related cancers. Curr Over Dise Health Res. 2022;4:71-111.
24. Safarpour P, Daneshi-Maskooni M, Vafa M, Nourbakhsh M, Janani L, Maddah M, et al. Vitamin D supplementation improves SIRT1, Irisin, and glucose indices in overweight or obese type 2 diabetic patients: a double-blind randomized placebo-controlled clinical trial. BMC Fam Pract. 2020;21(1):26.
25. Chang E, Kim Y. Vitamin D ameliorates fat accumulation with AMPK/SIRT1 activity in C2C12 skeletal muscle cells. Nutrients. 2019;11(11):806.
Download attachments: 10.4328.ACAM.22073
Hanan Salih Abdulrraziq, Hala Mahmoud Ibrahim Abou Heif, Hala Mohamed El-Sayed Maklad, Gehan Yassin Soliman Shoeib, Mona Hassan Fathelbab. Effect of moderate exercise and vitamin D3 on adipose tissue SIRT1 Protein in obese rats.Ann Clin Anal Med 2024;15(3):199-203
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
Our initial evaluation and clinical approaches in penetrating thoracic traumas
Fatoş Kozanlı 1, Ahmet Karslıgil 2
1 Department of Thoracic Surgery, Faculty of Medicine, Kahramanmaras Sutcu Imam University, 2 Department of Thoracic Surgery, Kahramanmaras NecipFazil City Hospital, Kahramanmaras, Turkey
DOI: 10.4328/ACAM.22081 Received: 2023-12-21 Accepted: 2024-02-12 Published Online: 2024-02-21 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):204-209
Corresponding Author: Fatoş Kozanlı, Department of Thoracic Surgery, Faculty of Medicine, Kahramanmaras Sutcu Imam University, 46040, On iki şubat, Kahramanmaraş, Turkey. E-mail: opdrfatoskozanli@gmail.com P: +90 505 223 19 84 Corresponding Author ORCID ID: https://orcid.org/0000-0001-7664-2657
This study was approved by the Ethics Committee of Kahramanmaraş Sütçü İmam University, Faculty of Medicine (Date: 2023-03-14, No: 04)
Aim: In this study, we aimed to explain the etiology of penetrating chest injuries, demographic characteristics of the injured, accompanying organ injuries, treatment, length of hospital stay, intensive care and blood replacement requirements, mortality and morbidity.
Material and Methods: The data of 130 patients over 18 years who applied to our institution with penetrating thoracic trauma were analyzed retrospectively. The age, gender, injury mechanism and instruments, localization of injury, affected organs, first and second interventions, length of stay, and the requirement for intensive care and blood transfusion were obtained from the hospital’s electronic database. Additionally, complications and mortality rates were recorded.
Results: In cases with a gunshot injury, thoracotomy was the first intervention, and this was statistically significant compared to the intervention in ordinary cut injuries (p<0.05). Pneumothorax, hemothorax, contusion, pulmonary laceration, diaphragm, liver, and spleen wounds were significantly higher in intensive care patients. Pneumothorax, hemothorax, contusion, pulmonary laceration, pneumo-mediastinum, localization of diaphragm, liver, spleen, colon, and gastric wounds were significantly higher in individuals with a hospital stay of ≥7.
Discussion: In our, the average age and gender distribution were similar to the literature. The most common injury mechanism in our study was incision wounds. Our study is similar to the literature in this respect. Our pneumothorax rates were quite high compared to previous literature. This may be attributed to the involvement of iatrogenic injuries. In our study, it was observed that all iatrogenic injuries resulted in pneumothorax. Our tube thoracostomy rates are quite high compared to the conservative treatment approach. The low rate of conservative approach in cases brought by penetrating trauma has been one of the factors contributing to survival.
Keywords: Penetrating Thoracic Trauma, Thorax, Hemothorax, Pneumothorax, Lung Laceration
Introduction
Penetrating thoracic traumas are divided into two groups: penetrating injuries and injuries with firearms. These injuries account for 30% of all traumas [1]. On the other hand, firearm injuries have high pressure resulting in severe damage to the surrounding tissue [2]. Penetrating thoracic traumas constitute 30% of all traumas [3]. Some chest wounds are life-threatening emergencies often associated with intrathoracic injury [4, 5]. Most low-velocity penetrating injuries do not require surgical treatment. Hence, tube thoracostomy is sufficient [6, 7]. Asymptomatic patients can be followed up with chest x-rays or computerized tomography [8]. However, sternotomy or thoracotomy may be required due to extensive pulmonary injury, intrathoracic major vascular injuries, and cardiac injuries[9].
Within the scope of this study, it was aimed to elucidate the risk factors affecting penetrating chest injuries, their etiology, demographic characteristics of the injured, severity of injury, accompanying organ injuries, treatment options, length of hospital stay and need for intensive care. and transfusion requirements, morbidity, and mortality.
Material and Methods
The data of 130 patients who were admitted to Kahramanmaraş Sütçü İmam University Faculty of Medicine and Kahramanmaraş Necip Fazıl City Hospital due to penetrating chest trauma between January 2016 and January 2023 were retrospectively examined.
Iatrogenic causes and penetrating thoracic traumas caused by any foreign body were also included in the study. These were included in the sharps injuries group. Cases with blunt thoracic trauma and those under the age of 18 were excluded from the study.
The intervention performed when we first accepted the case was considered the first intervention, and the interventions that were needed later during the follow-up or performed by other surgical departments were considered the second intervention.
The groups that needed intensive care and those that did not need intensive care were compared in terms of mortality, morbidity and complications. The patients who are injured by gunshots were compared with those injured by sharp objects.
The age, gender, injury mechanism and instruments, localization of injury, affected organs, first and second interventions, length of stay, and the requirement for intensive care and blood transfusion were obtained from the hospital’s electronic database. Additionally, complications and mortality rates were recorded. Mortalities were those that occurred during hospital follow-up.
Statistical Analysis
Patient data were analyzed with the IBM Statistical Package for the Social Sciences (SPSS) for Windows 28.0 program. Frequency and percentage were given as descriptive values for categorical data, and mean and standard deviation were given for continuous data. The distribution of variables was measured by the Kolmogorov-Smirnov test. Mann-Whitney-U test was used for quantitative independent variables, “the Pearson Chi-Square Test” was used to compare categorical variables, and the Fischer test was applied when the Chi-Square test conditions were not met.
Results were considered statistically significant when the P value was less than 0.05.
Ethical Approval
This study was approved by the Ethics Committee Kahramanmaraş Sütçü İmam University, Faculty of Medicine (Date: 2023-03-14, No: 04).
Results
Within this research, 130 patients with penetrating thoracic trauma were analyzed retrospectively. Of the cases, 109 (83.8%) were male and 21 (16.2%) were female. The mean age of the participants was 35.2±17.8 years. The mechanism of injury was a cutting tool in 88 (67.7%) cases. The most common type of thoracic injury was pneumothorax in 102 (78.5%) cases. Diaphragmatic injury was seen in 22 (16.9%) of the cases. Injury mechanism and injury types are given in Table 1. Pneumothorax, hemothorax, contusion, pulmonary laceration, diaphragm, liver, and spleen wounds were significantly higher in intensive care patients (p<0.05). The need for an initial intervention was significantly lower (p<0.05), while thoracotomy and tube intervention were significantly higher (p<0.05) in intensive care patients. As expected, the complication rate, mechanical ventilation, and blood transfusion requirements were significantly higher in the intensive care unit patients (p<0.05). The parameters in the groups with and without intensive care requirements are given in table 2. Rib fractures, soft tissue injury, pulmonary contusion, pulmonary laceration, subcutaneous emphysema, pneumo-mediastinum, and extremity wounds were significantly higher in the group with gunshot injury than in the other group (p<0.05).
In cases with a gunshot injury, thoracotomy was the first intervention, and this was statistically significant compared to the intervention in ordinary cut injuries (p<0.05). The tube intervention was significantly lower in gunshot wound patients (p<0.05).
Mechanical ventilator requirement and blood transfusion rate were significantly higher in the gunshot injury group (p<0.05) (Table 3).
Discussion
Penetrating thoracic traumas are more common in the younger age group although they are also relatively high in the second and fourth decades. The mean age of the subjects in this research was 35.2±17.8 years, which is similar to the literature [10]. Penetrating thoracic traumas are common in men since the use and supply of firearms can be accessed more easily compared to women. In a study by Lundin et al., the proportion of men was very high in our study. In our study, the injury rate of men was approximately five times higher than that of women [11].
Penetrating thoracic traumas occur with gunshot and stab wounds. These ratios may differ according to the geographies, regions, and occupations in the literature. In our study, the most common injury mechanism in the patients (67.7%) was cut wounds. Firearm gunshot wounds are expected to be higher in the military compared to other individuals [12]. Robison et al. reported the distribution of firearm gunshot wounds and penetrating traumas as 32.8% and 67.2%, respectively. Our study is similar to the literature in this respect [13].
Pneumothorax, as in published articles, was the most common type of intrathoracic injury in our analysis. However, our pneumothorax rates were quite high (78.5%) compared to the previous literature. This rate has been reported as 23.5% by Karamustafaoglu et al. and 44% by Kong et al. [14,15]. This may be attributed to the inclusion of iatrogenic injuries. In our study, it was observed that all iatrogenic injuries resulted in pneumothorax.
Various extremity injuries have been reported in our series within extrathoracic injuries. These injuries have been described in a wide range from skin injuries where primary repair is sufficient to extremity injuries requiring bone fixation. Gunshot wounds resulted in injuries leading to limb fractures. Due to the high kinetic energy of gunshot injuries, it has been observed that large bones cause fractures that require surgical treatment. Similar to the literature, bone fractures were also observed in our series [16].
Loogna et al. stated that liver and diaphragm injuries were the most common intra-abdominal organ injuries causing penetrating thoracic traumas. In addition, Demetriades et al. indicated that diaphragm injury was the most frequent injury type accompanying penetrating thoracic traumas . In our study, the most common intra-abdominal organ injuries were diaphragm injuries [17,18]. The incidence of these accompanying extrathoracic pathologies varies between 30 – 35%. Injuries to these organs cause perfusion and oxygenation disorders throughout the body, leading to morbidity and mortality [19]. In our study, extrathoracic organ injuries negatively affected morbidity and length of hospital stay.
As a result of the initial evaluation in our study, tube thoracostomy was the most commonly applied approach. Compared to a study conducted in our country, our tube thoracostomy rates are quite high compared to the conservative treatment approach [20]. The low rate of conservative approach to cases brought with penetrating trauma has been one of the factors contributing to survival.
With our mortality rate in our study was significantly different from the literature and was found to be %3.8. To the best of our knowledge, the lowest mortality rate reported in the literature is in our study. One of the reasons for this result is that we act more aggressively, especially in left hemithorax and parasternal injuries. In the literature, it is mentioned that more care should be taken in left hemithorax injuries. Injuries, especially to the left front of the chest, should be treated as if they were a heart injury until proven otherwise [21]. In addition, in our study, thoracotomy was preferred more in gunshot wounds due to the high kinetic energy of the bullet and the unpredictability of its path in the thorax. This preference is one of the reasons why our mortality is low.
In autopsy studies, 96.4% of the deaths in thoracic traumas were due to bleeding. The morbidity and mortality in cases with multiple organ trauma were increased compared to cases with isolated thoracic trauma. The causes of mortality were multiple organ dysfunction, acute respiratory distress syndrome, pulmonary embolism, crush syndrome, brain edema, sepsis, disseminated intravascular coagulation, and cerebrovascular events [22]. In our study, two of the five mortal cases were in the firearm injury group. The other two were patients followed in the intensive care unit due to multiple organ failure. These patients were in the iatrogenic injury group. Deaths in the gunshot injury group were due to massive hemorrhage and deaths in the iatrogenic injury group were due to multiple organ failure.
In our study, we preferred surgical intervention instead of conservative treatment, especially in gunshot injuries. Therefore, our thoracotomy rates are high in gunshot wounds. In addition, we performed surgical procedures for left hemithorax or parasternal region injuries regardless of the injury type. As a result, our mortality rates were significantly lower than in the literature [23, 24]. We have kept our intensive care indications and intensive care indications in penetrating chest trauma wider than the literature [25]. With this approach, we detected our patients who had a second surgery and needed blood and blood products earlier.
Conclusion
While most stable patients with penetrating thoracic traumas of civilian origin can be treated simply and successfully with initial chest tube placement, these patients should be evaluated comprehensively and systematically for the presence of life-threatening occult conditions. Stable patients should be closely monitored and repeatedly examined until study completion because there is potential for rapid decompensation and death due to initially unrecognized intrathoracic injuries. The presence of heart and large vessel injuries should be taken into consideration in gunshot wounds, injuries to the anterior chest wall and parasternal area.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: None
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Bautista-Parada IR, Bustos-Guerrero AM. Prevalencia de lesiones torácicas y abdominales en pacientes con traumatismo toracoabdominal penetrante [Prevalence of thoracic and abdominal injuries in patients with penetrating thoracoabdominal trauma]. Cir Cir. 2022;90(3):379-384.
2. Stretch B, Kyle A, Patel M. Management of penetrating chest trauma in the context of major trauma networks. Br J Hosp Med (Lond). 20222;83(10):1-7.
3. Yan T, Xie W, Xu M. Effect of chest ultrasound compared with pericardial window for the diagnosis of occult penetrating cardiac wounds in hemodynamically stable subjects with penetrating thoracic trauma: A meta-analysis. Int Wound J. 2023;20(6):2483-2491.
4. Liu A, Nguyen J, Ehrlich H, Bisbee C, Santiesteban L, Santos R, et al. Emergency Resuscitative thoracotomy for civilian thoracic trauma in the field and emergency department settings: A Systematic review and meta-Analysis. J Surg Res. 2022;273:44-55.
5. Vasse M, Leone M, Boyer L, Michelet P, Goudard Y, Cardinale M, et al de Lesquen H. Impact of the implementation of a trauma system on compliance with evidence-based clinical management guidelines in penetrating thoracic trauma. Eur J Trauma Emerg Surg. 2023;49(1):351-360.
6. Alshehab D, Al Ghunaim E, Al Otaibi FH, Aleid Y, Al Elewah A, Sotis R, et al. Role of uniportal video-assisted thoracoscopic surgery in the management of penetrating thoracic injuries. Kardiochir Torakochirurgia Pol. 2022;19(3):155-157.
7. Morgan B, Prakash K, Mayberry JC, Brickley MB. Thoracic trauma: Clinical and paleopathological perspectives. Int J Paleopathol. 2022;39:50-63.
8. Polireddy K, Hoff C, Kinger NP, Tran A, Maddu K. Blunt thoracic trauma: Role of chest radiography and comparison with CT – findings and literature review. Emerg Radiol. 2022;29(4):743-755.
9. Lodhia JV, Eyre L, Smith M, Toth L, Troxler M, Milton RS. Management of thoracic trauma. Anaesthesia. 2023;78(2):225-235.
10. Lundin A, Akram SK, Berg L, Göransson KE, Enocson A. Thoracic injuries in trauma patients: epidemiology and its influence on mortality. Scand J Trauma Resusc Emerg Med. 2022;12;30(1):69.
11. Williams BM, Mulima G, Charles A. Chest trauma management in low- and middle-income countries. Thorac Surg Clin. 2022;32(3):329-336.
12. Robison PD, Harman PK, Trinkle JK, Grover FL. Management of penetrating lung injuries in civilian practice. J Thorac Cardiovasc Surg. 1988;95(2):184-90.
13. Karamustafaoğlu YA, Yavaşman İ, Kuzucuoğlu M, Mammedov R, Yener Y. Penetran travmalı olgularda 13 yıllık deneyimimiz. [13-Year Experience with Penetrating Trauma Patients] Balkan Medical Journal. 2009;2009(3):232-236.
14. Kong VY, Sartorius B, Clarke DL. The selective conservative management of penetrating thoracic trauma is stil appropriate in the current era. Injury. 2015;46(1):49-53.
15. Krämer S, Graeff P, Lindner S, Walles T, Becker L. Okkulter und residueller Hämatothorax nach thorax trauma – empfehlungen der interdisziplinären arbeits gruppe thorax trauma der sektion NIS der Deutschen Gesellschaft für Unfall chirurgie (DGU) und der Deutschen Gesellschaft für Thorax chirurgie (DGT) [Occult and Retained Haemothorax – Recommendations of the Interdisciplinary Thoracic Trauma Task Group of the German Trauma Society (DGU – Section NIS) and the German Society for Thoracic Surgery (DGT)]. Zentralbl Chir. 2023;148(1):67-73.
16. Loogna P, Bonanno F, Bowley DM, Doll D, Girgensohn R, Smith MD, et al. Emergency thoracic surgery for penetrating, non-mediastinal trauma. ANZ J Surg. 2007;77(3):142-5.
17. Demetriades D, Velmahos GC. Penetrating injuries of the chest: Indications for operation. Scand J Surg. 2002;91(1):41-5.
18. Kumar A, Shiwalkar N, Bhate S, Keshavamurthy S. Management of thoracic and cardiac trauma: A Case Series and Literature Review. Cureus. 2022;30;14(6).
19. Tekinbaş C, Eroğlu A, Kürkçüoğlu İC, Türkyılmaz A, Yekeler E, Karaoğlanoğlu N. “Chest trauma: Analysıs of 592 cases.”Turkısh journal of trauma & emergency surgery. 2003;9(4):275-280.
20. Spiegl UJ, Osterhoff G, Bula P, Hartmann F, Scheyerer MJ, Schnake KJ, et al. Concomitant injuries in patients with thoracic vertebral body fractures-a systematic literature review. Arch Orthop Trauma Surg 2022;142(7):1483-1490.
21. Floan GM, Calvo RY, Prieto JM, Krzyzaniak A, Patwardhan U, Checchi KD, et al. Pediatric penetrating thoracic trauma: Examining the impact of trauma center designation and penetrating trauma volume on outcomes. J Pediatr Surg. 2023;58(2):330-336.
22. Beloy V, Dull M. Blunt chest wall trauma: Rib fractures and associated injuries. JAAPA. 2022;35(11):25-31.
23. Becker L, Schulz-Drost S, Schreyer C, Lindner S. Thorax drainage beim thorax trauma – Empfehlungen der interdisziplinären Arbeits gemeins chaft Thorax trauma der Deutschen Gesellschaft für Thorax chirurgie (DGT) und der Deutschen Gesellschaft für Unfall chirurgie (DGU) [Chest tube in thoracic trauma – recommendations of the interdisciplinary thoracic trauma task group of the German society for thoracic Surgery (DGT) and the German Trauma Society (DGU)]. Zentralbl Chir. 2023;148(1):57-66.
24. Sun J, Ren K, Zhang L, Xue C, Duan W, Liu J, et al Traumatic blunt thoracic aortic injury: A 10-year single-center retrospective analysis. J Cardiothorac Surg. 2022;17(1):335.
Download attachments: 10.4328.ACAM.22081
Fatoş Kozanlı, Ahmet Karslıgil. Our initial evaluation and clinical approaches in penetrating thoracic. Ann Clin Anal Med 2024;15(3):204-209
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
In vitro characterisation of antioxidant and anti-inflammatory activities of ceratonia siliqua L. extracts and synthesised zinc oxide nanoparticles
Emrah Caylak 1, Gokhan Nur 2
1 Department of Biochemistry, Faculty of Medicine, Girne American University, Kyrenia, Cyprus, 2 Department of Biomedical Engineering, Faculty of Engineering and Natural Sciences, Iskenderun Technical University, Hatay, Turkey
DOI: 10.4328/ACAM.22109 Received: 2024-01-15 Accepted: 2024-02-24 Published Online: 2024-02-28 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):210-215
Corresponding Author: Emrah Caylak, Department of Biochemistry, Faculty of Medicine, Girne American University, Kyrenia, Cyprus. E-mail: emrah333@hotmail.com P: +90 544 613 49 99 Corresponding Author ORCID ID: https://orcid.org/0000-0003-0408-9690
Aim: This study aimed to investigate the phytochemical contents of extract of the Carob plant, which grows and is endemic in Cyprus, and to evaluate the antioxidant and anti-inflammatory activities of synthesised zinc oxide nanoparticles/Ceratonia siliqua L.
Material and Methods: Phytochemical analysis of Ceratonia siliqua L. extract was performed, and total flavonoid, phenolic and alkaloid contents were determined. Then, ZnO nanoparticles/Ceratonia siliqua L. were synthesised using zinc acetate and characterised using various techniques such as UV, FTIR, XRD and SEM. Hydroxyl radical, 2,2-diphenyl-1-picryl hydrazyl, and 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) scavenging activities were used to determine as an antioxidant marker. To investigate the anti-inflammatory effect of Ceratonia siliqua L. extracts and ZnO-NPs/CsL, we performed assays such as heat-induced hemolysis, proteinase inhibitory activity and albumin denaturation tests.
Results: Our study results also revealed that Ceratonia siliqua L. extracts and synthesized ZnO-NPs showed a high radical capture potential and quite strong anti-inflammatory activity in vitro.
Discussion: Our study has shown that ZnO nanoparticles obtained by the green synthesis method have the potential to be used in a wide range of areas in medicine and will contribute to the literature in medical and technological terms.
Keywords: Antioxidant, Anti-Inflammatory, Ceratonia Siliqua L., Green Synthesis, ZnO Nanoparticles
Introduction
Many scientific studies have extensively investigated the antioxidant and anti-inflammatory effects of compounds such as phenolic, terpenoid, and ketone. With the development of bio-compatible, bio-safe and environmentally friendly nanomaterials, nanotechnology will be widely used in medicine in the future [1-3]. Green biosynthesis of metal nanoparticles such as zinc oxide nanoparticles (ZnO-NPs) using different plant extracts is a new option without producing toxic chemicals [4]. Carob (Ceratonia siliqua L.) is native to the Mediterranean countries of Turkey, Greece, Portugal, Spain, Italy and Cyprus [5]. Free radicals are produced during normal physiological cell metabolism in the body. However, when they occur in large quantities, they disrupt the proper functioning of cellular functions, damage cell components, and cause chronic diseases [6, 7]. The urges taken in by humans through their diet and fruit contain antioxidant compounds. Ceratonia siliqua L. extract has been shown to contain high amounts of antioxidant phytochemicals, including phenols, tannins and flavonoids [6]. This herb is well known to have antioxidant activity for free radicals [8].
Inflammation is a complex biological response of body tissues to adverse stimuli such as infections, damaged cells or irritants. It is characterised by the production of cytokines involved in the regulation of inflammatory reactions. Atherosclerosis and cancer are associated with inflammatory diseases that have been linked to excessive chronic production of inflammatory cytokines. Numerous pieces of evidences have revealed that Ceratonia siliqua L. extracts exhibit anti-inflammatory effects by inhibiting myeloperoxidase activity, human neutrophil reactive oxygen species (ROS) production, and other inflammatory mediators [8].
Ceratonia siliqua L. has anti-inflammatory, anti-microbial and antioxidant properties [6,8], and the synthesis of nanoparticles of this plant has yet to be thoroughly studied. Therefore, this study aimed to evaluate the determination of the phytochemical contents (total flavonoid, phenolic and alkaloid contents) of the extract of Ceratonia siliqua L. and to characterise green synthesised ZnO nanoparticles/Ceratonia siliqua L. (ZnO-NPs/CsL) using various techniques such as UV, FTIR, XRD and SEM. Then, we searched the extract of Ceratonia siliqua L. and synthesised ZnO-NPs/CsL for their antioxidant and anti-inflammatory activities.
Material and Methods
Carobs were collected from the hills of Alsancak and Lapta in the Kyrenia district, Cyprus. Ceratonia siliqua L., examined in this study, was dried in the shade for two weeks, and the pulp and seeds of carobs were powdered with an electric plant grinder. All chemicals and zinc acetate dihydrate (Zn(CH3COO)2.2H2O) to be used for the study were supplied by Merck Company (Germany).
Preparation of Ceratonia siliqua L. aqueous extract (CAE)
Firstly, 200 g of powdered Ceratonia siliqua L. was put into the balloon flask and boiled for 15 min at 80–90°C by adding 1 L of distilled water. Then, the mixture was cooled and filtered with a Whatman 1 filter paper. The extracts were stored at +4°C for research.
Phytochemical analysis of CAE
The total flavonoid content of CAE was determined using the aluminium chloride colorimetric method [9]. Initially, 0.2 mL of CAE was placed in a conical flask, 2 mL of distilled water and 750 μL of sodium nitrite solution was added, and incubated for 6 min at room temperature. Then 150 μL of AlCl3.6H2O was added. Absorbance was measured at 510 nm using a Shimadzu UV 1800 Spectrophotometer (Japan). Total flavonoid content levels were made using a standard curve using the following equation based on the calibration curve: y=0.0049x+0.0047, R2=0.9935, where y was the absorbance, and x was the quercetin. Quercetin was used as the reference material, and the calculation was done in mg of quercetin per g of sample.
The total phenolic content of CAE was determined using the Folin-Ciocalteu colorimetric method [9]. Briefly, 0.2 mL of CAE was mixed with 0.2 mL of Folin-Ciocalteau reagent and 2 mL of sodium carbonate solution. It was incubated at room temperature for two hours. The absorbance of the resulting blue colour solution was measured at 750 nm spectrophotometrically. Gallic acid was used as the reference material, and the calculation was made in mg of gallic acid per g of sample.
The total alkaloid content of CAE was determined according to the alkaloid-bromocresol green (BCG) reaction method, resulting in a yellow-coloured product [9]. 200 µL of CAE was placed in a test tube and 2 N HCl was added to make the final volume 1000 µL. The mixture was thoroughly mixed, transferred to a separatory funnel, and washed thrice using 10 mL of chloroform. The solution was neutralised by adding 0.1 N sodium hydroxide. After 10 min, 5 mL of BCG solution and 5 mL of phosphate buffer were added. The mixture was shaken, and the complex formed was extracted with chloroform. The extracts were collected and diluted with chloroform. Its absorbance was measured at 470 nm spectrophotometrically. The results were calculated using the formula [alkaloids= (Abs−0.048)/0.021] and expressed as mg-CAE/mL.
Synthesis and analysis of ZnO-NPs/CsL
To synthesise ZnO-NPs/CsL, 100 mL of CAE and 100 mL of zinc acetate solution were added to the beaker and kept at room temperature for one hour. The solutions were continuously heated and mixed at 100×g for four hours. The colour of the mixture changed from yellow to brown, indicative of ZnO-NPs/CsL formation. Next, it was purified by centrifugation with distilled water at 10000×g, 20 min. Supernatants were discarded, nanoparticle pellets were collected, washed with distilled water, freeze-dried using Alpha 1-2 LDplus and stored at -80°C [10]. The absorbance of ZnO-NPs/CsL was characterised at 200-800 nm, spectrophotometrically. The structural characterisation and particle size examination of ZnO-NPs/CsL were done by Transmission Electron Microscopy (TEM) and Scanning Electron Microscopic (SEM) (Hitachi, Japan). In addition, the spectra properties of those were carried out by Fourier Transform Infrared Spectroscopy (FTIR) (Bruker, Germany) in the spectra range of 4000–400 cm-1. X-ray diffraction (XRD) of the ZnO-NPs/CsL was analysed using an XPert PRO diffractometer (Holland) in the 2Ɵ range of 20–80o [10].
Antioxidant analysis of the ZnO-NPs/CsL
The determination of hydroxyl radical scavenging activity is based on the degradation of 2-deoxy ribose by the hydroxyl radical produced by the Fenton system (Fe+3, Ascorbic acid, EDTA, H2O2). The 2-deoxy ribose was condensed with thiobarbituric acid (TBA) and then measured as TBARS [9]. 45 μL of sodium phosphate buffer, 15 μL of deoxyribose, 150 μL of FeSO4-EDTA, 15 μL of H2O2 and 525 μL of deionised water were added to 750 μL of CAE/or ZnO-NPs/CsL/or L-ascorbic acid (used as a control). The solution was kept in the incubator for four hours. 75 μL of trichloroacetic acid (TCA) and 75 μL of TBA were added, and the reaction was initiated by adding NaOH. Then, it was incubated in a water bath (90oC, 10 min), cooled, and centrifuged at 10000×g for 15 min. The supernatant was aspirated, and its absorbance was recorded at 520 nm spectrophotometrically. Percentage inhibition of hydroxyl radical was determined using the Eq-1: % Inhibition=[(Acontrol-Asample)/Acontrol]×100
To determine 2,2-diphenyl-1-picryl hydrazyl (DPPH) activity [11], from the stock solution, several doses of the sample (50-100-150-250-500 μg/mL) were prepared. 100 μL of DPPH solution was added to 100 μL of CAE/or ZnO-NPs/CsL/or L-ascorbic acid. The mixture was thoroughly vortexed and incubated in the dark (20oC, 30 min). The decrease in absorbance at 517 nm was measured spectrophotometrically to detect DPPH colour degradation. It was calculated using Eq-1, and from a concentration plot against %inhibition, a linear regression analysis was performed to determine the IC50 value (the sample concentration needed to remove 50% free radicals) [11].
The 2,2′-azino-bis (3-ethylbenzotiazolin-6-sulfonic acid)(ABTS+) test assesses the ability of antioxidants to clear the oxidative species produced by ABTS+. ABTS+ radicals are formed by a reaction between ABTS+ and the highly oxidising potassium persulfate. As the antioxidant scavenging potential increased, a decrease in absorbance was observed [11]. ABTS was dissolved in water (7 mM). ABTS radicals were produced by adding potassium persulfate (2.45 mM). It was incubated in the dark (20oC, 12-16 h). 1 mL diluted of ABTS+ solution was added to 10 μL of CAE/or ZnO-NPs/CsL/or L-ascorbic acid. After 6 min of incubation, the blue/green ABTS+ solution turned pale yellow and then discoloured; absorbance was measured at 734 nm spectrophotometrically. Percentage inhibition of ABTS+ was calculated using the Eq-1.
Anti-inflammatory analysis of the ZnO-NPs/CsL
In vitro, to investigate the anti-inflammatory effect of CAE or ZnO-NPs/CsL, we performed heat-induced haemolysis, proteinase inhibitory activity and albumin denaturation assays [12, 13]. Acetylsalicylic acid was used as a reference drug, and Eq-1 was used to calculate the %inhibition.
For erythrocyte suspension, whole blood was taken from a healthy person with a consent form by ethical rules. Blood with 0.9 % NaCl was centrifuged at 2000×g for 5 min and rinsed to remove the supernatant. The mixture was reconstituted as a 10% suspension by adding sodium phosphate buffer (10 mM/pH 7.4). For heat-induced haemolysis testing, 0.06 mL of CAE/or ZnO-NPs/CsL/or acetylsalicylic acid, 0.06 mL of blood cell suspension, and 2.93 mL of phosphate buffer were incubated in a water-bath (54°C), and then centrifuged again at 2000×g, 3 min; and its absorbance was measured at 540 nm spectrophotometrically.
In the proteinase inhibitor activity test, 0.06 mg of trypsin and 1 mL of Tris-HCl buffer (20 mM/pH 7.4) were added to 1 mL of CAE/or ZnO-NPs/CsL/or acetylsalicylic acid, and then incubated at 37°C/5 min. The mixture was incubated and centrifuged for another 20 min by adding 1 mL of casein (0.8%). 2 mL of 70% perchloric acid was added to the mixture, and its absorbance was measured at 210 nm spectrophotometrically.
To determine the inhibition of albumin denaturation, 0.2 mL of 1% bovine albumin and 3.8 mL of phosphate-buffered saline (pH 6.4) were added to 1 mL of CAE/or ZnO-NPs/CsL/or acetylsalicylic acid. It was incubated (at 37°C for 15 min and then at 70°C for 5 min). After cooling, its absorbance was measured at 660 nm spectrophotometrically.
Statistical Analysis
The data from antioxidant assays were subjected to descriptive statistics, expressed as x±SEM and analysed using One-Way ANOVA for statistical comparison of differences among means followed by Tukey’s test for pairwise comparisons and values of p<0.05 were considered statistically significant.
Ethical Approval
All the reagents used in this study were prepared, used, and disposed of according to laboratory guidelines and material safety. Since no animals/humans were used during the study, the university ethics committee stated there was no need to get approval from the ethics committee.
Results
CAE’s total flavonoid, phenolic and alkaloid contents were 289.26±2.5, 116.57±2.83, and 354.47±2.85, respectively. They were expressed as mg/100 g-CaE.
Our study used various techniques such as UV, FTIR, XRD, and SEM to control the synthesis of ZnO-NPs/CsL from CAE and characterise their catalytic activities. First, ZnO-NPs/CsL formation was analysed by UV spectroscopy. In general, the absorbance values of ZnO-NPs/CsL at 360 nm indicate that particles are synthesised [14]. The peak values in our study, 365 nm, supported ZnO-NPs/CsL synthesis (Figure 1A). The absorption peaks measured by FTIR spectroscopy refer to the tensile vibrations O-H (alkene), C=C, N-O, and C=O (carbonyl) bindings (Figure 1B), respectively. Metal oxides usually give vibration frequencies below 1000 cm−1, and their peaks in the spectrum between 470 and 600 cm−1 are indicated to belong to ZnO-NPs/CsL.
Zeta potential testing was used to assess the electrical charge on the surface of ZnO-NPs, and XRD analysis was used to reveal the crystal structure of the synthesised ZnO-NPs/CsL (Figure 2A). As shown in the figure, the peaks were shown with 2θ values of 35.46, 37.01, 43.52, 48.61, 52.39, 64.26, and 75.44. These peaks corresponded to (100), (002), (101), (102), (110), (103), and (200), respectively. Finally, the SEM images revealed particles with rounded morphologies and an average size of 200 μm (Figure 2B). The results determined for the hydroxyl radical and DPPH scavenging or ABTS+ activities of CAE and ZnO-NPs/CsL are presented in Table 1. It was observed that the maximum antioxidant activities were at 500 µg/mL. The %inhibitions of the hydroxyl radical, DPPH and ABTS+ scavenging of the CAE were 47.82±0.03%, 54.68±0.12%, and 65.58±0.02%, respectively. These inhibitions for ZnO-NPs/CsL were 41.17±0.06%, 44.52±0.21%, and 49.58±0.04%, respectively. They were determined for the standard L-ascorbic acid as 49.26±0.08%, 58.21±0.08%, and 69.42±0.05%, respectively. The %inhibition values for CAE and ZnO-NPs/CsL were significantly lower than that of standard L-ascorbic acid, indicating excellent free radical inhibition at lower concentrations. We performed heat-induced haemolysis to determine the anti-inflammatory effects of CAE and ZnO-NPs/CsL. We found that the IC50 values for acetylsalicylic acid (2.06±0.84 mg/mL), CAE (3.89±0.72 mg/mL), and ZnO-NPs/CsL (3.08±0.97 mg/mL). In the proteinase inhibitory activity test, we determined IC50 values for acetylsalicylic acid (2.21±0.57 mg/mL), CAE (3.57±0.31 mg/mL), and ZnO-NPs/CsL (2.96±0.82 mg/mL). When we performed the albumin denaturation inhibition, the IC50 values were found for acetylsalicylic acid, CAE and ZnO-NPs/CsL as 3.01±0.63 mg/mL, 4.51±1.21 mg/mL, and 3.67±1.05 mg/mL, respectively. The values were increased with the decrease in the concentration of our measured test agents (Table 2).
Discussion
We determined the phytochemical contents of CAE in our study. The phenolic compounds have been discovered to be the most abundant plant substances with antioxidant activity. The flavonoids also are thought to have an antioxidant effect through a sweeping or chelating process. Previous studies of the Carob plant have shown that it contains significant amounts of phenolics and flavonoids, while Carob also contains high amounts of alkaloids [1]. There are also many reports indicating the presence of alkaloids in Carob [15], which may explain its traditional therapeutic uses. Many studies have shown that these molecules are well-known for their antioxidant and anticancer properties [14].
In our study, ZnO-NPs/CsL were synthesised using zinc acetate as a precursor. They characterised their catalytic activities using techniques such as UV, FTIR, XRD and SEM. First, ZnO-NPs/CsL formation was analysed by UV spectroscopy. In general, the absorbance values of ZnO-NPs/CsL at 360 nm indicate that particles are synthesised [14]. The peak values in our study, 365 nm, supported ZnO-NPs/CsL synthesis. The absorption peaks measured by FTIR spectroscopy refer to the tensile vibrations O-H (alkene), C=C, N-O, and C=O (carbonyl) bindings, respectively. CAE contained the phenolic compound, allowing the detection of the presence of phytochemicals and functional groups by FTIR analysis ranging from 400-4000 nm [14]. Metal oxides usually give vibration frequencies below 1000 cm−1, and their peaks in the spectrum between 470 and 600 cm−1 are indicated to belong to ZnO-NPs/CsL. The peak recorded at about 1700 cm−1 in our study implies alkene (C=C) and carbonyl (C=O) stretching, while the peak recorded at 1570 cm−1 was assigned to the aromatic bending vibration of the alkane group (C-H). Zeta potential testing was used to assess the electrical charge on the surface of ZnO-NPs, and XRD analysis was used to reveal the crystal structure of the synthesised ZnO-NPs/CsL. When the obtained XRD peaks were examined, they were consistent with the literature data [16].
ZnO-NPs are unique molecules due to their oxidation state. They are positively charged on their surfaces due to the gap in their “d and f “ orbitals and show antioxidant properties by reducing and scavenging electrons from other molecules. ROS are generated in the body during normal cellular metabolism. ROS are highly reactive and cause rapid oxidation of target molecules that lead to lipid peroxidation in cells among many biochemical reactions [17], thus causing many diseases caused by oxidative stress such as neurological diseases, old age, cancer and heart disease [18, 11]. ZnO-NPs effectively bind free radical species, hydroxyl radical, hydrogen peroxide, superoxide anion radical, and singlet oxygen, reducing oxidative stress. Medicinal plants contain high amounts of antioxidant phytochemicals, including phenols, tannins and flavonoids, and medicinal plants have been observed to prevent lipid peroxidation due to synthesis with ZnO [19, 20]. The determinations of hydroxyl radical, DPPH, and ABTS scavenging activities have long been used as an antioxidant marker in studies related to oxidative stress and focused on investigating the antioxidant effect of plants. Plant extracts whose hydroxyl radical, DPPH and ABTS+ radical scavenging activities were determined to have remarkable antioxidant potential [21]. Darwish et al. [22] reported that CAE exhibited potent antioxidant activity that prevented the formation of DPPH, hydroxyl, and nitric oxide free radicals. They suggested that the scavenging activity of the CAE was often evaluated according to its IC50; the lowest IC50 value corresponds to the highest free radical scavenging activity. We suggested that the studied CAE and ZnO-NPs/CsL are potent scavengers of ROS radicals in vitro and, therefore, have high antioxidant activity, as evidenced by low IC50 values.
In vitro, to investigate the anti-inflammatory effect of CAE and ZnO-NPs/CsL, we performed heat-induced haemolysis, proteinase inhibitory activity and albumin denaturation assays. For the potential anti-inflammatory effect of the medicinal plant Ceratonia siliqua L., it is used in various diseases, including diabetes, especially in Anatolia (Turkey) and Cyprus. It exerts its anti-inflammatory effects in controlling protein denaturation and albumin denaturation by inhibiting enzymes such as serine proteinase, which is released from lysosomal granules of neutrophils and has documented that leukocyte proteinase plays an essential role in the development of tissue damage during inflammatory disorders such as rheumatoid arthritis and diabetes. The literature has also shown that CAE has anti-inflammatory potential because it contains phenolic compounds, which are used for their anti-inflammatory and analgesic effects, strengthening the immune system, relieving pain, and treating skin disorders and asthma [23-25].
Conclusion
Our study indicated that CAE contained phenolic compounds and possessed antioxidant activity. ZnO-NPs/CsL have been shown to have antioxidant effects by reducing biomolecular agents in the structure of the plant with radical quenching activity. They also have been found to have a potent anti-inflammatory agent that protects cells against inflammation. Therefore, it is thought that ZnO-NPs/CsL, whose production and consumption potential is increasing, can provide many advantages to humanity in the treatment of oxidative/inflammatory diseases and cancer. Therefore, further investigations need to be conducted to investigate the effect of CAE and ZnO-NPs/CsL.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: None
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Chiriac ER, Chitescu CL, Geană EI, Gird CE, Socoteanu RP, Boscencu R. Advanced analytical approaches for the analysis of polyphenols in plants matrices—a review. Separations. 2021;8:65-70.
2. Seo Y, Hwang J, Lee E, Kim YJ, Lee K, Park C, et al. Engineering copper nanoparticles synthesised on the surface of carbon nanotubes for anti-microbial and anti-biofilm applications. Nanoscale. 2018;10(33):15529-44.
3. Dhakal DR, Kshetri YK, Chaudhary B, Kim T-H, Lee SW, Kim BS, et al. Particle size-dependent anticorrosion performance of the Si3N4 nanoparticle incorporated electroless nip coating. Coatings. 2022;12(1):9-15.
4. Zhong L, Liu H, Samal M, Yun K. Synthesis of ZnO nanoparticles decorated spindle shaped graphene oxide for application in synergistic antibacterial activity. J Photochem Photobiol B. 2018;183:293-301.
5. Koday, Z. Geographical Features of the State of the Turkish Republic of Northern Cyprus. Ataturk Uni Turkiyat Aras Ens Der. 2015;2:2-20.
6. Alqudah A, Qnais EY, Wedyan MA, Oqal M, Alqudah M, AbuDalo R, et al. Ceratonia siliqua leaves ethanol extracts exert anti-nociceptive and anti-inflammatory effects. Heliyon. 2022;8(8):e10400-5.
7. Caylak E, Aytekin M, Halifeoglu I. Antioxidant effects of methionine, alpha-lipoic acid, n-acetylcysteine and homocysteine on lead-induced oxidative stress to erythrocytes in rats. Exp Toxicol Pathol. 2008;60(4-5):289-94.
8. Rtibi K, Jabri MA, Selmi S, Souli A, Sebai H, El-Benna J, et al. Carob pods (Ceratonia siliqua L.) inhibit human neutrophils myeloperoxidase and in vitro ROS-scavenging activity. RSC Adv. 2015;5:84207-15.
9. Sadiq MB, Hanpithakpong W, Tarning J, Anal AK. Screening of phytochemicals and in vitro evaluation of antibacterial and antioxidant activities of leaves, pods and bark extracts of Acacia nilotica (L.) del. Industrial Crops & Product. 2015;77:873-82.
10. Siby J, Beena M. Microwave-assisted green synthesis of silver nanoparticles and the study on catalytic activity in the degradation of dyes. J Mol Liq. 2015;204:184-91.
11. Thaipong K, Boonprakob U, Crosby K, Cisneros-Zevallos L, Byrne DH. Comparison of ABTS, DPPH, FRAP, and ORAC assays for estimating antioxidant activity from guava fruit extracts. J Food Compos Anal. 2006;19:669-75.
12. Yesmin S, Paul A, Tarannum N, Rahman A, Akhter SF, Wahed MII, et al. Membrane stabilisation as a mechanism of the anti-inflammatory activity of ethanolic root extract of choi (Piper chaba). Clin Phytoscience. 2020;6:1-10.
13. Gunathilake KDPP, Ranaweera KKDS, Rupasinghe HPV. Influence of boiling, steaming and frying of selected leafy vegetables on the ın vitro anti-inflammation associated biological activities. Plants (Basel). 2018;7(1):22-8.
14. Karmous I, Taheur FB, Zuverza-Mena N, Jebahi S, Vaidya S, Tlahig S, et al. Phytosynthesis of zinc oxide nanoparticles using Ceratonia siliqua L. and evidence of anti-microbial activity. Plants (Basel). 2022;11(22):3079-85
15. Lakkab I, El Hajaji H, Lachkar N, Lefter R, Ciobica A, El Bali B, et al. Ceratonia siliqua L. seed peels: Phytochemical profile, antioxidant activity, and effect on mood disorders. J Funct Foods. 2019;54:457-65.
16. Rajeshkumar S, Parameswari RP, Sandhiya D, Al-Ghanim KA, Nicoletti M, Govindarajan M. Green synthesis, characterisation and bioactivity of Mangifera indica seed-wrapped zinc oxide nanoparticles. Molecules. 2023;28(6):2818-24.
17. Farmer EE, Mueller MJ. ROS-mediated lipid peroxidation and RES-activated signalling. Ann Rev Plant Biol. 2013;64:429-50.
18. Caylak E, Halifeoglu I. Effects of sulfur-containing antioxidants on malondialdehyde and catalase levels of liver, kidney and brain in lead-exposed rats. J Med Sci. 2007;27(1):1-8.
19. Bajpai VK, Sharma A, Kang SC, Baek KH. Antioxidant, lipid peroxidation inhibition and free radical scavenging efficacy of a diterpenoid compound sugiol isolated from Metasequoia glyptostroboides. Asian Pac J Trop Med. 2014;7(1):9-15.
20. Xu DP, Li Y, Meng X, Zhou T, Zhou Y, Zheng J, et al. Natural antioxidants in foods and medicinal plants: extraction, assessment and resources. Int J Mol Sci. 2017;18(1):96-105.
21. Fidrianny I, Budiana W, Ruslan K. Antioxidant activities of various extracts from Ardisia Sp. leaves using DPPH and CUPRAC assays and correlation with total flavonoid, phenolic, carotenoid content. Int J Pharmacogn Phytochem Res. 2015;7:859-65.
22. Darwish WS, Khadr AS, Kamel MA, Abd Eldaim MA, El Sayed ET, Abdel-Bary HM, et al. Phytochemical characterisation and evaluation of biological activities of Egyptian carob pods (Ceratonia siliqua L.) aqueous extract: in vitro study. Plants (Basel). 2021;10(12):2626-34.
23. Bañuls C, Rovira-Llopis S, Falcón R, Veses S, Monzó N, Víctor VM, et al. Chronic consumption of an inositol-enriched carob extract improves postprandial glycaemia and insulin sensitivity in healthy subjects: A randomised controlled trial. Clin Nutr. 2016;35(3):600-7.
24. Rico D, Martín-Diana AB, Martínez-Villaluenga C, Aguirre L, Silván JM, Dueñas M, et al. In Vitro approach for evaluation of carob by-products as source bioactive ingredients with potential to attenuate metabolic syndrome (Mets). Heliyon. 2019;5(1):e01175-82.
25. Benchikh Y, Louailèche H, George B, Merlin A. Changes in bioactive phytochemical content and in vitro antioxidant activity of Carob (Ceratonia siliqua L.) as influenced by fruit ripening. Indust Crops & Product. 2014;60:298-303.
Download attachments: 10.4328.ACAM.22109
Emrah Caylak, Gokhan Nur. In vitro characterisation of antioxidant and anti-inflammatory activities of ceratonia siliqua L. extracts and synthesised zinc oxide nanoparticles. Ann Clin Anal Med 2024;15(3):210-215
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/
Increased carotid intima media thickness as an indicator of increased cardiovascular risk in patients with primary familial erythrocytosis
Alpay Yeşilaltay 1, Hasan Değirmenci 2
1 Department of Hematology and Tumor Immunology, Faculty of Medicine, Baskent University İstanbul Hospital, 2 Department of Cardiology, Şişli Etfal Training and Research Hospital, İstanbul, Turkey
DOI: 10.4328/ACAM.22129 Received: 2024-01-30 Accepted: 2024-02-28 Published Online: 2024-02-29 Printed: 2024-03-01 Ann Clin Anal Med 2024;15(3):216-220
Corresponding Author: Alpay Yeşilaltay, Department of Hematology and Tumor Immunology, Faculty of Medicine, Baskent University Istanbul Hospital, 34662, İstanbul, Turkey. E-mail: alpay.yesilaltay@hotmail.com P: +90 507 240 42 84 Corresponding Author ORCID ID: https://orcid.org/0000-0003-3784-5739
This study was approved by the Ethics Committee of Namık Kemal University, Faculty of Medicine (Date: 2015-04-09, No: 2015/48)
Aim: Erythrocytosis (polycythemia) is defined in males as Hb (hemoglobin) level >16.5 g/dL and Hct (hematocrit) level >49%, while in females, it is defined as Hb level >16.0 g/dL and Hct level >48%. It is classified into two groups based on EPO (erythropoietin) levels: primary and secondary. In the primary group, EPO levels are normal, while in the secondary group, elevated EPO levels are observed. Those carrying JAK2 mutations in the primary group are diagnosed with Polycythemia Vera, whereas those without mutations are defined as familial erythrocytosis. In our study, we aimed to evaluate potential increased cardiovascular risks in patients with primary familial erythrocytosis (PFE).
Material and Methods: A total of 64 patients diagnosed with primary familial erythrocytosis, characterized by normal EPO levels and the absence of JAK2 mutations, were included in the study, along with 30 healthy volunteer controls. The lipid parameters of both groups were examined. Additionally, carotid intima-media thickness (CIMT), considered an indicator of subclinical atherosclerosis in both groups, was measured using non-invasive carotid Doppler ultrasonography (USG).
Results: The lipid parameters of both groups were examined, and no statistically significant differences were detected between the groups. However, in the measurement of carotid intima-media thickness (CIMT), a statistically significant increase was observed in the PFE (primary familial erythrocytosis) group compared to the control group in both carotid arteries.
Discussion: Patients with PFE (Primary Familial Erythrocytosis) being considered as polycythemic but not classified as hematological patients often lead to occasional disruptions in the follow-up and treatment of patients. As demonstrated in our study, PFE patients are prone to increased atherosclerosis and related complications independent of lipid parameters. Considering the hereditary nature of the disease and its onset, particularly in young age, it is important to monitor patients using methods such as Carotid Doppler USG and CIMT, with the belief that this could prevent cardiovascular complications that may occur especially at early ages
Keywords: Carotid Intima, Cardiovascular Risk, Erythrocytosis, JAK2 Mutations
Introduction
Erythrocytosis is a group of disorders frequently encountered in haematology practice. Erythrocytosis (polycythemia) is defined as elevated haemoglobin (Hb) and/or haematocrit ratio (Hct) in peripheral blood. This ratio is defined as an Hb value >16.5 g/dL in males and >16.0 g/dL in females and an Hct value >49% in males or >48% in females [1].
Erythrocytosis is basically grouped as primary and secondary according to EPO (Erythropoietin) level, and then both groups are divided into two groups as hereditary and acquired. EPO level is normal in the primary form. Primary Familial Erythrocytosis (PFE) form often involves EPO mutations (germline mutations). Mutations in EPO receptors result in increased erythrocyte production despite physiological EPO levels [2]. This is hereditary and often has a family history of early cardiovascular and cerebrovascular disease events. Primary acquired polycythaemia is Polycythaemia Vera (PV), which involves (somatic mutations; clonal) (JAK2 mutations). Here, JAK mutations are mutations of the JAK2V617F or Exon 12 region. PV is a chronic myeloproliferative disorder involving the bone marrow, with a risk of leukaemia and myelofibrosis [3].
The basic rule in the causes of secondary polycythaemia is elevated EPO. Secondary hereditary type includes germline mutations (VHL, EGLN1, EPAS) and methaemoglobinaemia. Acquired secondary polyschaemia is mainly due to hypoxic causes. In this group of patients, pulmonary, cardiac, endocrine, high altitude and renal transplantation are the main causes.
In the approach to polycythemic patients in haematology polyclinics, if the patient’s EPO level is normal, the patient does not have JAK mutations and there are no secondary causes of polycythemia, the patient is considered as PFE, and the patients are followed up with intermittent phlebotomies. Although PFE does not have the risk of haematological malignancy, cardiac and cerebral events at an early age in family members are frequently encountered in the anamnesis of patients. For this purpose, we wanted to evaluate the possible cardiovascular risk in the PFE group and measured carotid intima- media thickness (CIMT) with high-resolution B-mode carotid ultrasonography, which is known to be a suitable method for detecting subclinical atherosclerosis.
Carotid Doppler ultrasonography (USG) is one of the most sensitive and reliable methods for high- resolution morphological evaluation of carotid arteries. Since it is non-invasive, it is one of the most important advantages of the method and the detected CIMT increase is accepted as an early morphological finding of atherosclerosis. In addition to morphological information, colour and spectral Doppler USG has the advantage of showing the flow changes caused by vascular lesions in real time.
The patient’s position should be supine, head hyperextended, neck neutral or at a 30-45° angle to the side opposite to the side being evaluated. It is performed using high-resolution, linear aligned probes for examination. The frequency range of the probes is 5-18MHX, and they are designed for vascular purposes.
The examination consists of 3 steps:
1. Real-time B-mode/grey-scale imaging: (axial and longitudinal): It is used for general morphological evaluation. CIMT is used to detect the presence of plaque and to evaluate the plaque structure. CIMT should normally be smaller than 0.8 mm. Exceeding this value is considered to increase the risk of cardiovascular and cerebrovascular events [4].
2. Colour Doppler US (CDUS) (Axial and Longitudinal): A critical level common carotid artery stenosis proximal to the axial axis and the associated external-internal carotid artery steal phenomenon are sensitive in indicating common carotid artery dissection or occlusion, whereas longitudinal axis-CDUS is used to assess whether previously detected plaque-level stenoses cause aliasing.
3. Spectral Doppler US (longitudinal): The most important criterion for determining the presence and severity of carotid artery stenosis is flow velocity assessment [5].
Therefore, we compared Carotid Doppler USG and CIMT of polycythemic patients admitted to the Haematology and Cardiology outpatient clinics of our hospital with a healthy control group in order to evaluate the underlying atherosclerosis and to determine their current cardiovascular risks.
Material and Methods
Patients:
Our study was conducted jointly by the haematology and cardiology clinics of Tekirdağ Namık Kemal University between January 2014 and January 2017. Inclusion criteria included patients with isolated erythrocytosis (haemoglobin [Hb] >18.5g/dL in males or >16.5g/dL in females) and patients with high Hb values. JAK2 V617F and Exon 12 mutation analyses were performed and those who were negative in both tests were included in the study. Serum EPO levels were measured in all patients and patients with normal EPO levels were included in the study. Patients with high EPO levels were excluded. Patients with known severe pulmonary disease (Chronic Obstructive Pulmonary Disease-Bronchial Asthma) and severe cardiovascular disease, known coronary artery disease, atrial fibrillation, atrial flutter, congenital heart disease, history of cardiac operation, congestive heart failure, left bundle branch block were also excluded, and patients under 18 years of age and patients with known cerebrovascular disease or peripheral arterial disease were excluded.
Sixty-four patients evaluated as PFE and 30 healthy individuals constituting the control group were included in the study. After comprehensive clinical evaluations of the patients and the control group, a survey was applied to those who agreed to participate in the study and the patients were questioned in terms of erythrocytosis. In addition, routine laboratory results (blood count and lipid parameters) were recorded. Cardiovascular risk factors (diabetes, smoking, hypertension and lipids) were determined in patients and controls. Written informed consent was obtained from all subjects.
Statistical evaluation:
SPSS for Windows 18.0 software was used to evaluate the statistical analysis. Shapiro-Wilk test was used to examine whether the data were normally distributed. All variables were found to be normally distributed. Continuous variables were expressed as mean ± standard deviation and categorical variables were expressed as number and percentage. In the statistical analysis, continuous variables between the control and patient groups were compared by Student t test. Chi-square tests were used to compare categorical variables. Pearson correlation analysis was performed to investigate the correlation between values less than .05 and was considered statistically significant in genetic terms.
CIMT measurements:
CIMT measurements were performed after the participants rested for 15 min in the supine position with their heads tilted backwards. The right and left carotid arteries were imaged by an experienced cardiologist using a high-resolution B-mode ultrasound device (GE Vivid S5: General Electric VingMed Systems, Horten, Norway) with a 12L-RS broadband linear transducer. Right and left common carotid arteries were visualised in the longitudinal plane. Measurements were made manually by determining a 1cm segment from 2 cm below the carotid bulb. 3 measurements were averaged. Carotid plaques were not included in the measurement.(Figure:1)
Ethical Approval
This study was approved by the Ethics Committee of Namık Kemal University, Faculty of Medicine (Date: 2015-04-09, No: 2015/48).
Results
A total of 90 people, 64 patients and 30 controls were included in the study. The mean age was 46 years in both groups and there was no age difference between the groups. Again, the gender distribution of males and females was equal and similar to each other. The rates of concomitant hypertension and diabetes were similar in patients and controls in both groups, and there was no difference between the groups. Smoking rates were the same in both groups. Lipid profiles were similar in the form of Triglycerides, HDL-C, LDL-C, Total Cholesterol and there was no statistically significant difference between the two groups for all lipid parameters. The mean Hb level in the patient group was 18.1 g/dL and Hct level was 58.2%, while the mean Hb level in the control group was 14.5 g/dL and Hct level was 43.9%, and there was a statistically significant difference between the groups for both values (Table 1).
CIMT values of the patients were determined as follows. Both CIMT (Right and Left Carotid Intima Media Thickness) were found to be higher in the patient group. Carotid intima-media thickness was significantly increased in the patient groups compared to the control group. A statistically significant difference was found between the groups. This difference was found in both carotid arteries (Table 2)(Figure: 2)
Discussion
Unlike Polycythemia Vera, which is a chronic myeloproliferative disease with polycythemia, PFE does not progress to any haematological disease and does not involve the bone marrow. Therefore, since it is not a disease of the bone marrow and does not cause a secondary haematological disease, it may be ignored in haematology outpatient clinics from time to time. However, in cardiology outpatient clinics, patients tend to be referred to haematology outpatient clinics because of the polycythemic picture present in the patient. As a result, this group of patients may actually be stuck in the middle in terms of follow-up and treatment.
In the analyses we performed in patients presenting with polycythemia in our outpatient clinic follow-up, we observed that this group of patients with negative JAK2 V617F and Exon 12 mutations, no secondary polycythemia and normal EPO values were predominantly male. In retrospective anamnesis, we also observed that sudden death, acute coronary syndrome and cerebrovascular events, which usually occur at young ages, were frequent in this group of patients, especially in first-degree family relatives. Therefore, we planned to investigate this issue to determine the possible increase cardiovascular risk in PFE patients. In our literature review, we observed that the number of studies on PFE is not very high. We did not find any cardiovascular risk studies, especially in this group of patients in our literature review. More frequently, case reports were available.
Since one of the problems that arise in the diagnosis of PFE is the uncertainty and inadequacy in the mutations defining this group of diseases, the diagnosis is based on the exclusion of other secondary polycythaemia and JAK2(+) myeloproliferative diseases. Since PFE-defining mutations are not well characterised, it is not frequently used in clinical practice. These mutations mostly involve amino acid changes in Exon 8 of the EPO receptor gene [6]. However, since the identified mutations cannot be demonstrated in many cases, the term Idiopathic Familial Polycythaemia is also used instead of PFE in some sources [7].
Since cardiac risk studies directly evaluating PFE patients have not been performed, we wanted to evaluate and interpret our study by examining similar studies performed in other groups with polycythemia. Chuvas polycythemia, which is a similar type of polycythemia found in an endemic region in Asia, is an endemic polycythemia with partially similar features to PFE. Chuvas polycythaemia has a proven cardiovascular risk and increased thrombus formation [8]. Premature death and cardiovascular events have been reported in people living in this region, especially in male individuals. An increase in the incidence of cancer was also investigated in this group, but no risk was found [9].
It is accepted that many mechanisms increase cardiovascular events in polycythemia and that this triggers underlying atherosclerosis independently of lipids. One of these views is that moderate hypoxia triggers pulmonary hypertension by increasing ventilation in this group of patients, which may lead to an increased cardiovascular risk. In the same study, increased endothelin-1 levels were found in this group of patients and increased systolic pulmonary pressure was detected by Doppler ECHO in these patients [10]. Endothelin-1 is known to have high vasoconstriction properties in the arterial system. These mechanisms are likely to be present not only in PV but also in all polycythaemias and PFE. In fact, the data of our study support that there is increased atherosclerosis in PFE patients, although there is no difference between lipid parameters in the patient and control groups. Atherosclerosis will lead to thromboembolism and acute coronary and cerebrovascular events
In one of the limited studies conducted on this subject, thromboembolic events observed especially at young ages were associated with PFE [11]. There are case reports linking AEs with EPOR mutation to haemorrhage, DVT, coronary artery disease and MI, though milder in patients with EPOR mutation [12]. It has been shown in some publications and case reports that AE can also lead to abortion in women[13]. Considering all of these, prophylaxis is recommended in all cases of PFE [2]. The possible cause of abortion has been attributed to a thrombus in the placenta.
Haematocrit has the strongest influence on whole blood viscosity. One unit increase in haematocrit can cause up to 4% increase in blood viscosity. As haematocrit increases, blood flow velocity in the vessels slows down considerably due to viscosity due to increased resistance [14]. It is suggested that increased cardiac workload due to increased viscosity as a result of increased erythrocyte mass in polycythemia, increased endothelial shear and endothelial dysfunction contribute to atherosclerosis. Increased viscosity resulting from increased erythrocyte count may lead to hypoperfusion, hypoxia and tendency to coagulation [15]. Another study has also shown that PV is associated with endothelial dysfunction [16]. Myocardial performance index, which indicates systolic and diastolic dysfunction, was found to be impaired for both right and left ventricles in PV. Although studies have not been performed in PFE patients, a similar effect in PFE patients is due to the increased viscosity that occurs rather than the pathogenesis of the disease. In our TÜBİTAK study, we detected diastolic dysfunction in the same group of patients by echocardiography. We attributed this dysfunction to increased cardiac workload due to increased viscosity in PFE patients. In our clinical observations, we observed that cardiovascular events occurred more frequently and severely as Hct ratios increased.
One mechanism that has emerged in recent years involves a study in patients with PV that examined neutrophil-to-lymphocyte ratio (NLR) and carotid plaque burden in patients with essential thrombocythaemia and PV. It was emphasised in the study that a significantly higher rate of carotid plaque was observed in both groups of patients compared to healthy controls [17]. The point to be explained here is to show how a lipid-independent atherosclerosis occurs. Although it is known that elevated white blood cell (WBC) levels are not a direct factor in vascular damage, atherosclerotic plaque development, rupture and thrombosis, the correlation between inflammation and MI was proposed more than 50 years ago. Since then, a great deal of evidence has emerged supporting the important role of inflammation in coronary artery disease (CAD) and other manifestations of atherosclerosis [18].
In this context, it was accepted that immune cells are the dominant cells in early atherosclerotic lesions and accelerate the progression of lesions with their effector molecules. It was stated that direct activation of inflammation can lead to Acute Coronary Syndrome [19]. Neutrophils are presumed to mediate plaque rupture and thrombosis by secreting proteolytic enzymes that cause vascular damage mediated by the secretion of pro-inflammatory cytokines, activation of coagulation pathways, microvascular occlusion, and myocyte necrosis [20].
Similar mechanisms similar to PV in PFE patients, physiological stress and subsequent activation of the neurohormonal system may lead to cortisol release, mediating increased neutrophil count and relative lymphopenia through apoptosis. Thus, the neutrophil- to-lymphocyte ratio (NLR) may serve as a combined marker for both reactive and adaptive components of the inflammatory response as a result of increased neutrophil counts. All these mechanisms are likely to play a role in the atherosclerosis in PFE patients as in PV.
In our study, we compared current PFE patients with the normal population. Our study was designed prospectively. There was no statistically significant difference between both groups in all lipid sub-parameters in lipid profile examinations. When selecting the control group, the characteristics of the patient group were taken into consideration and care was taken to ensure that they had the same characteristics.
We evaluated the increase in CIMT (carotid intima media) thickness as an indicator of increased cardiovascular risk using high-resolution B-mode ultrasound. CIMT determination with carotid Doppler USG can be used non-invasively as a warning by showing IMT (intima-media thickness) in the early period. Measurement of carotid intima-media thickness by B-mode carotid ultrasonography is a suitable method to detect subclinical atherosclerosis. Carotid intima-media thickness greater than 0.9-1 mm is likely to be indicative of atherosclerosis and increased risk of cardiovascular disease [21]. We found significant and statistical increase in CIMT in the patient group compared to the control group. This increase was interesting because although the lipid profiles were similar in both groups of patients, the carotid intima was thicker in the PFE group. This was evident in the whole group with PFE.
Conclusion
As a conclusion of our study, PFE is a disease that requires joint follow-up in haematology and cardiology outpatient clinics. We believe that it is important for the protection of future generations that family examinations are carried out. We consider that screening of family members is important for preventing future complications and preventive medicine in PFE patients beyond the identification of a possible risk of cardiovascular disease in the patient alone.
Scientific Responsibility Statement
The authors declare that they are responsible for the article’s scientific content including study design, data collection, analysis and interpretation, writing, some of the main line, or all of the preparation and scientific review of the contents and approval of the final version of the article.
Animal and Human Rights Statement
All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards.
Funding: None
Conflict of Interest
The authors declare that there is no conflict of interest.
References
1. Swerdlow SH, Campo E, Harris NL, Jaffe ES, Pileri SA, Stein H, Thiele J.Editors. WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues WHO Classification of Tumours. Lyon. 2017.p.39.
2. Finazzi G, T Gregg X, Barbui T, Prchal JT. Idiopathic erythrocytosis and other non-clonal polycythemias. Best Pract Res Clin Haematol. 2006;(19):471–82.
3. Cross NC. Genetic and epigenetic complexity in myeloproliferative neoplasms. Hematology Am Soc Hematol Educ Program. 2011;(2011):208–14.
4. Den Ruijter H, Peters SA, Anderson T, Britton A, Dekker J, Eijkemans M, et al. Common carotid intima-media thickness measurements in cardiovascular risk prediction: a meta-analysis. JAMA. 2012; 308(8): 796-803.
5. Van den Oord S, Sijbrands EJ, Kate GL, Klaveren D, Domburg R, Steen A. et al. Carotid intima-media thickness for cardiovascular risk assessment: Systemic review and meta-analysis. Atherosclerosis. 2013;228(1):1-11.
6. Pasquier F, Marty C, Balligand T, Verdier F, Grosjean S, Gryshkova V, et al. New pathogenic mechanisms induced by germline erythropoietin receptor mutations in primary erythrocytosis. Haematologica. 2017.103;(4):575-86
7. Vočanec D, Prijatelj T, Debeljak N, Kunej T. Genetic variants of erythropoietin (EPO) and EPO receptor genes in familial erythrocytosis. Int J Lab Hematol. 2019;41(2):162-67.
8. Gordeuk VR, Prchal JT. Vascular complications in Chuvash polycythemia. Semin Thromb Hemost. 2006;32(3):289–94.
9. Gordeuk V, Sergueeva A, Miasnikova G, Okhotin D, Voloshin Y, Choyke P, et al. Congenital disorder of oxygen sensing: Association of the homozygous Chuvash polycythemia VHL mutation with thrombosis and vascular abnormalities but not tumors. Blood. 2004;103(10):3924–32.
10. Bushuev V, Miasnikova G, Sergueeva A, Polyakova L, Okhotin D, Gaskin P, et al. Endothelin-1, vascularendothelial growth factor and systolic pulmonary artery pressure in patients withChuvash polycythemia. Haematologica. 2006;91(6):744–49.
11. Huang LJ, Shen YM, Bulut GB. Advances in understanding the pathogenesis of primary familial and congenital polycythaemia. Br J Haematol. 2010;148(6):844-52.
12. Vočanec D, Prijatelj T, Debeljak N, Kunej T. Genetic variants of erythropoietin (EPO) and EPO receptor genes in familial erythrocytosis. Int J Lab Hematol. 2019;41(2):162-167.
13. Wu L, Sun R, Liu Y, Liu Z,Chen H, Shen S, et al. High hemoglobin level is a risk factor for maternal and fetal outcomes of pregnancy in Chinese women: A retrospective cohort study. BMC Pregnancy Childbirth. 2022;22(1):290
14. Finazzi G. Idiopathic atherosclerosis and other non- clonal polycythemias. Best Pract Res Clin Haematol. 2006;(19):471–82.
15. Kayrak M, Acar K, Gul EE, Bağlıcaklıoğlu M, Kaya Z, Sonmez O, et al. Assessment of left ventricular myocardial performance by tissue Doppler echocardiography in patients with polycythemia vera. Echocardiography. 2011;28(9):948-54.
16. Kralovics, R. and Prchal, JT. Genetic heterogeneity of primary familial and congenital polycythemia. Am J Hematol. 2001;68(2):115-21.
17. Neunteufl T, Heher S, Stefenelli T, Pabinger I, Gisslinger H, et al.Endothelial dysfunction in patients with polycythaemia vera.Br J Haematol. 2001;115(2):354-9.
18. Krečak I, Lucijanić M. Platelet-to-lymphocyte ratio and accelerated atherosclerosis in essential thrombocythemia and polycythemia vera. Nutr Metab Cardiovasc Dis. 2022;32(8):2010-11.
19. Bursi F, Weston S, Killian JM, Gabriel SE, Jacobsen SJ, Roger VLJ. C-reactive protein and heart failure after myocardial infarction in the community. Am J Med. 2007;120(7):616-22.
20. Hansson GK. Inflammation atherosclerosis and coronary artery disease N Engl J Med. 2005;352(16):1685-95.
21. Huang Y, Rabb H, Womer KL. Ischemia–reperfusion and immediate T cell responses Cell Immunol.2007;248(1):4-11
22. Amato M, Montorsi P, Ravani A, Oldani E, Galli S, Ravagnani SP, et al. Carotid intima-media thickness by B-mode ultrasound as surrogate of coronary atherosclerosis; Correlation with quantitative coronary angiography and coronary intravascular ultrasound findings. Eur Heart J. 2007;28(17):2094-101.
Download attachments: 10.4328.ACAM.22129
Alpay Yeşilaltay, Hasan Değirmenci. Increased carotid intima media thickness as an indicator of increased cardiovascular risk in patients with primary familial erythrocytosis. Ann Clin Anal Med 2024;15(3):216-220
Citations in Google Scholar: Google Scholar
This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/