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Martina Vukoja, Marina Ćurlin, Katarina Vukojević, Nevenka Jelić-Knezović, Anita Kolobarić, Martina Orlović Vlaho, V. Šoljić

This study aimed to investigate the cytotoxic activity of decidual lymphocytes and the mRNA/protein expression of cytotoxic proteins in various cell types in the context of preeclampsia (PE) compared to those of healthy pregnancies. We analyzed fresh decidua basalis tissue and tissue embedded in paraffin (FFPE) from PE pregnancies (n = 15) and compared them with those of healthy pregnancies (n = 15) of the corresponding gestational age. Using double immunofluorescence staining, we observed differences in the intensity and distribution of staining for granzyme K (GZMK) and FasL in extravillous trophoblasts. RT-qPCR analysis of FFPE placental tissue showed that GZMK mRNA expression was statistically higher (p < 0.0001) in PE compared to that of healthy controls. On the contrary, there was a low expression (p < 0.001) of FasL mRNA in PE compared to controls, while there was no statistically significant difference for IFN-γ mRNA between PE and controls. Although the level of cytotoxic activity changed depending on the ratio of effector and target cells, there was no significant difference observed between PE and controls in this in vitro study. In conclusion, in PE, extravillous trophoblasts exhibited increased expression of GZMK and decreased expression of FasL. These changes may contribute to impaired trophoblast invasion. However, these alterations did not appear to affect the cytotoxic properties of decidual lymphocytes. Additionally, the possibility of cell sorter separation of decidual lymphocytes would greatly contribute to a better understanding of single cells’ genetic profiles.

Deni Ramljak, Martina Vukoja, Marina Ćurlin, K. Vukojević, Maja Barbaric, U. Glamočlija, Bejana Purisevic, Olivera Perić et al.

Healthy and controlled immune response in COVID-19 is crucial for mild forms of the disease. Although CD8+ T cells play important role in this response, there is still a lack of studies showing the gene expression profiles in those cells at the beginning of the disease as potential predictors of more severe forms after the first week. We investigated a proportion of different subpopulations of CD8+ T cells and their gene expression patterns for cytotoxic proteins (perforin-1 (PRF1), granulysin (GNLY), granzyme B (GZMB), granzyme A (GZMA), granzyme K (GZMK)), cytokine interferon-γ (IFN-γ), and apoptotic protein Fas ligand (FASL) in CD8+ T cells from peripheral blood in first weeks of SARS-CoV-2 infection. Sixteen COVID-19 patients and nine healthy controls were included. The absolute counts of total lymphocytes (p = 0.007), CD3+ (p = 0.05), and CD8+ T cells (p = 0.01) in COVID-19 patients were significantly decreased compared to healthy controls. In COVID-19 patients in CD8+ T cell compartment, we observed lower frequency effector memory 1 (EM1) (p = 0.06) and effector memory 4 (EM4) (p < 0.001) CD8+ T cells. Higher mRNA expression of PRF1 (p = 0.05) and lower mRNA expression of FASL (p = 0.05) at the fifth day of the disease were found in COVID-19 patients compared to healthy controls. mRNA expression of PRF1 (p < 0.001) and IFN-γ (p < 0.001) was significantly downregulated in the first week of disease in COVID-19 patients who progressed to moderate and severe forms after the first week, compared to patients with mild symptoms during the entire disease course. GZMK (p < 0.01) and FASL (p < 0.01) mRNA expression was downregulated in all COVID-19 patients compared to healthy controls. Our results can lead to a better understanding of the inappropriate immune response of CD8+ T cells in SARS-CoV2 with the faster progression of the disease.

V. Šoljić, Maja Barbaric, Martina Vukoja, Marina Ćurlin, Martina Orlović Vlaho, E. Černi Obrdalj, Lidija Lasić Arapović, Daniela Bevanda Glibo et al.

Simple Summary CD8+ T cells are prominent decidual cells in the third trimester of healthy human pregnancy. They have a cytotoxic capacity which may control invasion of extravillous trophoblast and therefore affect placentation and play the role in development of preeclampsia. In this study, we examined the expression of CD8+ T cells in decidual tissue and peripheral blood of women with severe and mild preeclampsia in comparison to gestational age-matched healthy pregnancies. Additionally, the expression of cytotoxic proteins in CD8+ T cells was examined in order to specify their subpopulations. Abstract In our study, we aimed to establish expression of cytotoxic CD8+ T cells in the decidua basalis and the maternal peripheral blood (mPBL) of severe and mild preeclampsia (PE) and compare to healthy pregnancies. Decidual tissue and mPBL of 10 women with mild PE, 10 women with severe PE, and 20 age-matched healthy pregnancy controls were analyzed by double immunofluorescence and qPCR, respectively. By double immunofluorescence staining, we found a decreased total number of cells/mm2 in decidua basalis of granulysin (GNLY)+ (p ˂ 0.0001), granzyme B (GzB)+(p ˂ 0.0001), GzB+CD8+(p ˂ 0.0001), perforin (PRF1)+ (p ˂ 0.0001), and PRF1+CD8+ (p ˂ 0.01) in the severe PE compared to control group. Additionally, we noticed the trend of lower mRNA expression for GNLY, granzyme A (GZMA), GzB, and PRF1 in CD8+ T cells of mPBL in mild and severe PE, with the latter marker statistically decreased in severe PE (p ˂ 0.001). Forkhead box P3 (FOXP3) mRNA in CD8+ T cells mPBL was increased in mild PE (p ˂ 0.001) compared to controls. In conclusion, severe PE is characterized by altered expression of cytotoxic CD8+ T cells in decidua and mPBL, suggesting their role in pathophysiology of PE and fetal-maternal immune tolerance.

Ivona Ljevak, I. Vasilj, Josip Lesko, M. Neuberg, Olivera Perić, Marina Ćurlin

SUMMARY Providing in-patient nursing care inevitably involves shift work and shift patterns have been identified as an important factor in determining well-being and satisfaction among nurses. Shifts of 12 h or longer have become increasingly common for nurses in hospitals in some European countries. Longer shifts offer a potential to benefit from a compressed working week, with fewer work days and more days off-work, lower commuting costs, and increased flexibility. Most people find shift work less desirable compared to standard working hours. A cross-sectional cohort study was conducted at the Mostar University Hospital in 2019. A total of 157 subjects participated in the study, 22 (14%) of which were male (medical technicians) and 135 (86%) female (nurses). The mean age of study subjects was 33.3 years (min=20, max=54, SD=8.033). Results of this study suggested that nurses working irregular rotating shifts, with more family members to look after and more than 18 years of professional experience were affected by sleep disturbances between shifts. The study showed that shift work nurses had significantly higher levels of cortisol and prolactin compared with first-shift nurses/medical technicians. The results of this study are expected to stimulate further studies of sleep disturbances among shift nurses.

Marijana Marković-Boras, A. Causevic, Marina Ćurlin

Background: Coronary artery disease as a consequence of atherosclerosis is the most common cause of morbidity and mortality in type 2 Diabetes Mellitus (DM) patients. Homocysteine (HCY), as one of the risk factors, and uric acid (UA) as the most common antioxidant in serum have their roles in the processes of inflammation and atherogenesis, which underlie the pathogenesis of acute myocardial infarction (AMI). The effect of HCY in cardiovascular disease is thought to be manifested primarily through oxidative damage, implying a potential correlation between the HCY level and antioxidant status. Since the data related to the diagnostic significance of both HCY and UA in diabetic patients with AMI are conflicting, and so far not reported in Bosnian patients, this research aimed to examine the association of HCY and UA levels with glomerular filtration rate (eGFR) and explore the pathophysiological significance of these data in Bosnian diabetic patients with AMI. Methods: This prospective research included 52 DM type 2 patients diagnosed with AMI. Blood samples were taken on admission and used for biochemical analysis. Results of the biochemical analyses were statistically analysed. Results: Elevated HCY and UA levels were observed in diabetic patients. Females have higher HCY compared to males. A positive correlation was revealed between HCY and UA and was confirmed with different HCY levels in subgroups with different UA level. A negative correlation was observed between UA and HbA1c, as well as between both HCY and UA with eGFR. Conclusions: These results contribute to the clarification of the biochemical mechanisms characteristic in AMI patients with DM. According to these results, we believe that joint measurement of HCY and UA could enable a better assessment of the prognosis for this group of patients. This kind of assessment, as well as regression analysis, can identify high-risk patients at an earlier stage when appropriate interventions can influence a better outcome in such patients.

R. Babić, Mario Babić, P. Rastović, Marina Ćurlin, Josip Simic, Kaja Mandić, Katica Pavlović

Zdenko Šarac, Ružica Zovko, Marina Ćurlin, P. Filaković

Dental health and mental health are strongly associated. Neglecting either of them can negatively influence on the other and induce many health and communication problems. Association between oral/dental health, self-esteem, quality of life and holistic health has been recognized for a long time. There has been increasing interest in dental health among patients with major mental disorders as well as in mental states and problems among patients with orodental disorders. Despite of huge progress in the field of dentistry psychiatric patients have had poor oral/dental health. Patients with major mental disorders have quite number of the risk factors for oral disease and consequently poorer dental health, but oral/dental problems and diseases are commonly overlooked or neglected. Bad or inappropriate dental care is related to the patients' amotivation, ignorance, fears, low economic status, stigmas and negative attitudes by the medical professionals. It is important to stress that dental diseases in psychiatric patients deserve the same attention as other comorbid somatic diseases. In this review we accentuate the need for more collaboration in order to bridge the professional gap between dentistry and psychiatry.

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