Faculty of Medicine

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UTERINE ARTERY DOPPLER AND SERUM LEVEL OF IMMUNOBIOMARKERS IN PREECLAMPSIA - OUR EXPERIENCE
(Македонско лекарско друштво = Macedonian medical association/De Gruyter, 2024)
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Paneva, Iva
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Introduction. Important mechanisms are known to be involved in the immunomodulatory pathways which are crucial for maintaining an adequate utero-placental circulation in pregnancy. Its disbalance brings to impaired tolerance, which leads to inflammation and autoimmune processes in preeclampsia. Aim. The aim of this study was to find if inadequate uteroplacental hemodynamic was associated with improper feto-maternal immune adaptation. The risk of developing preeclampsia can be predicted by combining use of uterine artery flow and cytokine values. The aim was to show their combination as a predictive indicator of preeclampsia in the second trimester of pregnancy. Methods. This study enrolled 96 pregnant patients in the second trimester (patients were between the 14th and 20th gestational weeks). Their history data, routine foetal ultrasound, bilateral uterine artery Doppler ultrasound and cytokines were evaluated. All patients were followed up till the end of pregnancy. Half of the pregnant women consisted the study group (N=48), which had presence of notch of the uterine artery. In the control group (N=48), there was an absence of uterine artery notch. In all patients, Doppler of the uterine artery, pulsatility index (PI) and resistance index (RI) were made and determined. The pro-inflammatory cytokines (TNF-α, IL-1α, IL-2 and IL-6) and antiinflammatory cytokines (IL-4 and IL-10) from patient’s serum were analyzed. Results and Discussion. In the study group (N=48), 32 patients had changes in the cytokine serum levels. Increased pro-inflammatory biomarkers (IL-6, TNF-α, IL-1α) were with sensitivity of 78 to 91.2%. According to this, high predictive value was found. Of these 32 patients, 21 developed preeclampsia. When the sensitivity of pro- and anti-inflammatory biomarkers together with the uterine artery Doppler ultrasound was combined, a sensitivity resulted in 81.5%. Multivariate regression analysis detected that IL-6 was the most significant predictive parameter. This result is similar to that presented in the study by Teran and Hentschke et al. Conclusion. Using predictive tests is imporatnt to detect undeveloped preeclampsia in a timely manner that would prevent possible developmental complications. After abnormal Doppler results, cytokines should be investigated as a predictive method.
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Correlation of serum levels of inflammatory cytokines with severe form of cholecystitis
(Faculty of Medicine, Ss. Cyril and Methodius University in Skopje, 2022)
Spasovski, Zharko
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Novevska Petrovska, Biljana
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Stojanoski, Sashko
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Introduction: Cholecystitis is an inflammatory response of the body triggered by a number of mutually supporting biological mechanisms, which by creating and releasing inflammatory mediators - cytokines activate the innate or acquired immune system, which leads to neutralization of the harmful stimulus and initiation of the process of repair and regeneration of damaged tissue or its continuation as a long-term chronic process with simultaneous tissue destruction and reparation. Material and methods: The study was conducted at the General City Hospital (GCH) "8th September" in Skopje and the Institute of Immunology and Human Genetics in Skopje, in the period of 2020-2022. Statistical analysis of the data was performed with the statistical package SPSS for Windows 26.0. Results: The study included 165 subjects with gallbladder inflammation divided into 3 groups: mild, moderate and severe inflammation grade. Patients with mild, moderate, and severe inflammatory processes differed significantly in IgG levels (p = 0.049), IgA (p = 0.021), and IgM (p = 0.016) and insignificantly in IgE1 levels (p = 0.16). Patients with a severe inflammatory process had a higher prevalence of IL-2R and IL-8 than patients with a mild grade (p = 0.035; p = 0.26, respectively). The intensity of inflammatory process had a nonsignificant effect on the levels of TNF-alpha (p = 0.078), and a significant effect on the levels of fibrinogen (p = 0.001), with significantly higher levels of fibrinogen in the group of patients with severe inflammatory process compared to the group with mild grade (p = 0.0009). Conclusion: The intensity of inflammatory process affects the serum levels of inflammatory cytokines with presence of strong correlation between the severe form of cholecystitis and elevated serum levels of certain inflammatory cytokines.
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The Role of Interleukin-1 and Interleukin-6 in the Development of Acute Myocardial Infarction
(2023-08-26)
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Elizabeta Srbinovska-Kostovska
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Slavica Josifovska
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Certain cytokines as interleukin-1 (IL-1) and interleukin-6 (IL-6) are produced by the neutrophils and macrophages localized in the atheromatous plaques. The levels of circulating inflammatory cytokines IL-1 and IL-6 are found to be significantly increased in patients with ST-Segment Elevation Acute Myocardial Infarction (STEMI). In a group of 38 patients with STEMI, the levels of IL-1ß and IL-6 were determined in the coronary circulation (CC) by analysis of aspirates from the culprit lesions and the levels of these inflammatory markers in the systemic (peripheral) circulation (SC) during STEMI and six months after STEMI. Coronary angiography (CA) and percutaneous coronary intervention (PCI) were performed in all patients and the levels of cytokines were determined by the ELISA method. By examining the levels of IL-1ß and IL-6, the aim of this study was to determine their predictive value in short-term prognosis. In the acute phase (STEMI) there was a statistically significant difference between the mean values of IL-1ß (p=0, 000000) and IL-6 (p=0, 026204) in the samples of CC and SC, respectively. Results from the six months of follow-up showed that there were statistically no significant differences between the mean values of IL-1ß and IL-6 in the CC and SC. Moderate and low-positive correlation between the mean values of IL-1ß (r=0,6816; p=0,000) and IL-6 (r=0,4291; p=0,000) in CC and SC was observed at the sixth month of follow-up. In conclusion, our results confirm that IL-1ß and IL-6 are linked to the progression of Coronary Artery Disease (CAD) and should be considered as predictive markers.
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Oxidative stress index as prognostic marker for disease severity and its correlation with proinflammatory cytokines and lymphocyte subpopulation in COVID-19 patients
(2023-07)
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Introduction The ultimate goal of the SARS-CoV-2 virus is to learn how to evade the host's immune system . The pathogenicity of the virus, the comorbidities of infected individuals, and the ability of the host immune system to respond to induced cytopathic effects have a profound effect on the course and outcome of the disease. Our aim was to analyze several inflammatory, clinical laboratory parameters and oxidative stress markers and to provide comprehensive view of them for their future implementation in routine clinical practice. Method 35 patients, positive on SARS-CoV-2 were hospitalized at the University Clinic for Infectious Diseases and Febrile Conditions in Skopje. All patients were not vaccinated, since the study was performed before the onset of the national vaccination program. The total antioxidant capacity (PAT) and the plasma peroxide (d-ROMs) concentrations were performed on the FRAS5 analytical photometric system. The multi parameter flow cytometry was performed on full blood immediately after sample collection. Immunophenotyping was done by using BD FACSCanto™ II analyzer on lysed whole blood samples. For the simultaneous quantitative detection of multiple analytes from a single patient sample for IL-6 and VEGF we have used the High Sensitivity Evidence Investigator™ Biochip Array technology. Results Patients with moderate form of the disease had lower values of the measured concentration of dROMs and hence lower oxidative stress index when compared to the patients classified with the severe form of the disease (p=0.0001). We report a statistically significant difference of IL-6 and VEGF levels between the moderate and severe groups of patients (p=0.0001). We have observed decreased levels of absolute leukocytes count, CD45+ mononuclear and its subsets CD4+, CD8+, CD3+, NK cells (p<0.05). Also, CD19+ and CD45+ were decreased in the severe group in comparison to the group of patients with moderate COVID-19 (p>0.05). The oxidative stress parameters, OSI and d-ROM demonstrated a good correlation with CD45+ and CD4+. Additionally, only in the moderate group, we have obtained a good correlation amongst the investigated cytokines (IL-6 and VEGF) and NK cells was obtained (namely for IL-6, r=0.6973, p<0.05; and for VEGF, r=0.6498, p<0.05), whereas in the severe group only these cytokines correlated with CD45+ (for IL-6, r=0.5610, p<0.05; and for VEGF, r=0.5462, p<0.05). Conclusion The oxidative stress index can be used as a cheaper alternative and as a triage tool between severe and moderate illnesses, after showing good correlation with more expensive patient classification analysis
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Cardiovascular disease and COVID-19: a consensus paper from the ESC Working Group on Coronary Pathophysiology & Microcirculation, ESC Working Group on Thrombosis and the Association for Acute CardioVascular Care (ACVC), in collaboration with the European Heart Rhythm Association (EHRA)
(Oxford University Press (OUP), 2021-09-16)
Cenko, Edina
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Badimon, Lina
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Bugiardini, Raffaele
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Claeys, Marc J
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De Luca, Giuseppe
The cardiovascular system is significantly affected in coronavirus disease-19 (COVID-19). Microvascular injury, endothelial dysfunction and thrombosis resulting from viral infection or indirectly related to the intense systemic inflammatory and immune responses are characteristic features of severe COVID-19. Pre-existing cardiovascular disease and viral load are linked to myocardial injury and worse outcomes. The vascular response to cytokine production and the interaction between SARS-CoV-2 and ACE2 receptor may lead to a significant reduction in cardiac contractility and subsequent myocardial dysfunction. In addition, a considerable proportion of patients who have been infected with SARS-CoV-2 do not fully recover and continue to experience a large number of symptoms and post-acute complications in the absence of a detectable viral infection. This conditions often referred to as "post-acute COVID-19" may have multiple causes. Viral reservoirs or lingering fragments of viral RNA or proteins contribute to the condition. Systemic inflammatory response to COVID-19 has the potential to increase myocardial fibrosis which in turn may impair cardiac remodelling. Here we summarize the current knowledge of cardiovascular injury and post-acute sequelae of COVID-19. As the pandemic continues and new variants emerge, we can advance our knowledge of the underlying mechanisms only by integrating our understanding of the pathophysiology with the corresponding clinical findings. Identification of new biomarkers of cardiovascular complications, and development of effective treatments for COVID-19 infection are of crucial importance.
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Correlation of cervical length, fetal fibronectin, phIGFBP-1, and cytokines in spontaneous preterm birth up to 14 days from sampling
(De Gryter, 2014-12)
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Stefanovic, Milan
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Abstract Objective: The aim of this study was to determine the relationship between sonographic cervical length, fetal fibronectin (fFN), phIGFBP-1 (actim partus test), cytokines (IL-6, IL-2R, and TNF- α ), and spontaneous preterm birth (SPTB) up to 14 days from sampling. Methods: Fifty-eight patients were recruited in a period of 6 months from September 2013 until March 2014 with symptoms or complaints suggestive of preterm labor. Consenting women were treated according to usual hospital protocol, with addition of vaginal swabs taken for fetal fibronectin, phIGFBP-1 (actim partus test) and cervical IL6, IL2R, and TNF- α . The outcome variable was occurrence of preterm delivery within 14 days from the day of hospital admission. Results: Thirty-six patients (62.07%) were delivered within 14 days from admission. Our results indicated that the cervical length significantly inversely correlates with the concentration of IL-6 in the CVF (Spearman ’ s coefficient R = – 0.382, P < 0.05). Cervical length also correlated with a positive phIGFBP-1 test, i.e., patients with a positive test had an average cervical length of 18.5 ± 4.63 mm, which is significantly lower than patients with a negative test – 23.43 ± 7.39 mm (P = 0.003). Conclusions: The studied biochemical markers were only moderately successful in the prediction of preterm delivery.
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Correlation of the system of cytokines in moderate and severe preeclampsia
(IMR Press, 2016-01)
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M. Hadži-Lega
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M. Stefanovic
Objective of the study: To study the production of pro-inflammatory (IL-1β, IL- 2, IL-6, IL-8) and anti-inflammatory (IL-4, IL-10) cytokines in pregnancy complicated by preeclampsia in the third trimester. Institution: University Clinic of Gynecology and Obstetrics, Skopje, Republic of Macedonia. Material and Methods: Fifty women with pregnancies complicated by preeclampsia in the third trimester and 50 women with physiological pregnancy. Levels of IL-1β, IL- 2, IL-6, IL- 8, IL- 4, and IL- 10 were measured by using a solid -phase enzyme immunoassay. Statistical data processing was done using the application program SPSS for Windows 13, 0. To describe the distribution of analyzed variables, descriptive methods (mean, median, minim and max) were used . Results: In pregnancies complicated by preeclampsia, there are increased levels of proinflammatory cytokines and a change in the behaviour of opposing pools. Most pronounced changes in the levels of proinflammatory cytokines were observed in mild preeclampsia. In severe preeclampsia there was reduction of the concentration of anti-inflammatory cytokines IL- 4 and IL-10. Conclusion: The use of assessment cytokine profile monitoring of health status of women with preeclampsia is expedient.
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Correlation of the system of cytokines in moderate and severe preeclampsia
(IMR Press, 2016-01)
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Milan Stefanovic
Objective of the study: To study the production of pro-inflammatory (IL-1β, IL- 2, IL-6, IL-8) and anti-inflammatory (IL-4, IL-10) cytokines in pregnancy complicated by preeclampsia in the third trimester. Institution: University Clinic of Gynecology and Obstetrics, Skopje, Republic of Macedonia. Material and Methods: Fifty women with pregnancies complicated by preeclampsia in the third trimester and 50 women with physiological pregnancy. Levels of IL-1β, IL- 2, IL-6, IL- 8, IL- 4, and IL- 10 were measured by using a solid -phase enzyme immunoassay. Statistical data processing was done using the application program SPSS for Windows 13, 0. To describe the distribution of analyzed variables, descriptive methods (mean, median, minim and max) were used . Results: In pregnancies complicated by preeclampsia, there are increased levels of proinflammatory cytokines and a change in the behaviour of opposing pools. Most pronounced changes in the levels of proinflammatory cytokines were observed in mild preeclampsia. In severe preeclampsia there was reduction of the concentration of anti-inflammatory cytokines IL- 4 and IL-10. Conclusion: The use of assessment cytokine profile monitoring of health status of women with preeclampsia is expedient.
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Biochemical Indicators as Predictive Markers by Combining Clinical Signs in Pre-eclampsia
(ID Design 2012/DOOEL Skopje, 2020)
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Vlatko Girevski
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Abstract AIM: To determine whether previously identified risk factors are associated with the development of a severe form of pre-eclampsia in a heterogeneous cohort of women, and the predictive values of these risk factors when combined with certain biochemical indicators. MATERIALS AND METHODS: Systematic review of data collected for a doctoral case-control study plus an examination of the indicators of pre-eclampsia and maternal IL10 levels. This examination was conducted in 100 women with pregnancies complicated by varying degrees of pre-eclampsia and in 80 normotensive patients hospitalized at the University Clinic of Gynecology and Obstetrics, Skopje, Republic of Macedonia. Patients with preeclampsia were categorized into moderate (m PE) and severe (s PE) pre-eclampsia group according to the degree of pre-eclampsia. The severity of pre-eclampsia was determined according to the definition of the World Health Organization, Handbook for guideline development from 2010. RESULTS: The regression analysis applied in this study showed that elevated systolic blood pressure of 160 mmHg or higher, diastolic blood pressure of 100 mmHg or higher, pregnancy at an older age, nulliparity, persistent proteinuria in pregnancy, the serum lactate dehydrogenase concentration of 450 U/L or higher, and reduced serum concentrations of IL10 as significant predictors of severe pre-eclampsia in pregnant women. While other variables predicted a higher likelihood for the development of severe pre-eclampsia, IL10 decreased such likelihood. IL10 was also found to be negatively correlated with proteinuria and positively correlated with blood platelets. Significantly higher concentration of IL10 was confirmed in patients with a higher number of platelets in the blood and vice versa. On the other hand, the serum concentration of IL10 was significantly lower in patients with a higher amount of proteins in the urine and vice versa. CONCLUSIONS: Examination of clinical risk factors combined with biochemical markers can improve the predictive success of pre-eclampsia and has important clinical values in improving the prognosis of pregnant women and fetuses.
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Connection between cytokines and complications derived from preeclampsia pregnancies
(ScopeMed Publishing, 2020-05)
Preeclampsia is a condition of multiorgan involvement that can cause severe complications for the mother’s health and endanger the intrauterine development of the fetus. Purpose: The purpose is to detect the risk of developing preeclampsia in the second trimester, by examining cytokines and closely monitoring the pregnancy for complications from preeclampsia, whether they are affected by the same cytokines. Material and methods A total of 100 patients were monitored in the second trimester between 14 and 20 weeks of gestation. Values of immune biomarkers of their serum were analyzed after obtaining anamnestic data and performing ultrasound examination. With the help of the ELISA methodology, cytokines were verified: TNF-a, IL-1a, IL-2, and IL-6 versus IL-4 and IL-10. Results Of the 100 patients examined, 21 patients developed clinical symptoms and were diagnosed with preeclampsia in the third trimester. The interaction of proinflammatory interleukins is in favour of a mutual increase, and a decrease in the values of antiinflammatory interleukins is a significant predictive parameter in the second trimester for the development of preeclampsia. The increase in IL-6 is the largest statistically significant variable in the prediction of preeclampsia. The correlation between IL-6 and pregnancy complications is also with statistical significance (p=0.012). Conclusion The impaired immune response can result in consequences such as multiple organic disorders that occur in the clinical preeclampsia syndrome and problems with fetal development. The benefit of analyzing cytokines is highly significant for early diagnose and prevention of further complications.

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