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Aim This study aimed to assess the impact of forced repeated swimming stress on serum adiponectin and endothelin-1 levels in Wistar rats, comparing the effects between those fed a standard diet and those on a high-fat diet. Methods Twenty adult male Wistar rats were divided into two dietary groups: a standard food diet group (NFD, n=10) and a high-fat diet group (HFD, n=10). Both groups underwent daily forced swimming stress for six days, with durations increasing from 5 to 30 minutes. The protocol finished in an acute bout of swimming exercise on the seventh day with a duration of 40 minutes. Adherence to ethical guidelines was strictly maintained, and serum adiponectin and endothelin-1 levels were measured pre- and post-exercise using the ELISA method. Results Before swimming, the mean adiponectin levels were 4.30±1.50 ng/mL in the NFD group and 3.53±0.70 ng/mL in the HFD group. Post-exercise, these levels significantly decreased to 2.4±0.84 ng/mL (p=0.003) and 1.58±0.23 ng/mL (p=0.001), respectively. Endothelin-1 levels also showed significant decreases from 0.86 pg/mL (0.74-0.87) to 0.49 pg/mL (0.43-0.62) (p=0.003) in the NFD group, and from 0.89 pg/mL (0.86-0.93) to 0.69 pg/mL (0.60-0.75) (p=0.027) in the HFD group after swimming. Conclusion The study highlighted the significant effects of forced swimming stress on lowering serum adiponectin and endothelin-1 levels in Wistar rats, with more pronounced decreases observed in rats on a high-fat diet. The results of the study suggest the potential of exercise as a crucial component of strategies aimed at managing obesity and improving cardiovascular health, emphasizing the interaction between physical stress and dietary factors on metabolic and cardiovascular biomarkers.

C. Ladeira, Peter Møller, Lisa Giovannelli, G. Gajski, A. Haverić, E. Bankoglu, A. Azqueta, Marko Gerić et al.

Biomonitoring of human populations exposed to chemical substances that can act as potential mutagens or carcinogens, may enable the detection of damage and early disease prevention. In recent years, the comet assay has become an important tool for assessing DNA damage, both in environmental and occupational exposure contexts. To evidence the role of the comet assay in human biomonitoring, we have analysed original research studies of environmental or occupational exposure that used the comet assay in their assessments, following the PRISMA-ScR method (preferred reporting items for systematic reviews and meta-analyses extension for scoping reviews). Groups of chemicals were designated according to a broad classification, and the results obtained from over 300 original studies (n = 123 on air pollutants, n = 14 on anaesthetics, n = 18 on antineoplastic drugs, n = 57 on heavy metals, n = 59 on pesticides, and n = 49 on solvents) showed overall higher values of DNA strand breaks in the exposed subjects in comparison with the unexposed. In summary, our systematic scoping review strengthens the relevance of the use of the comet assay in assessing DNA damage in human biomonitoring studies.

The chemical composition of Pulicaria dysenterica(L.) Bernh. aerial parts essential oil (EO), growing wildin Bosnia and Herzegovina, was presented in the study. In addition to the EO composition, antimicrobial and cytotoxic activities were also tested. The aerial parts of P. dysentericacontained 0.3% of yellow, liquid, fragrant EO. The 51 components identified accounted for 81.09% of the oil. The EO was characterized by the presence of a high concentration of oxygenated sesquiterpenes 51.83% while oxygenated monoterpenes constituted 15.57%, sesquiterpene hydrocarbons 9.32% and non-terpene compounds presented 4.37% of the EO. The dominant compounds were the sesquiterpenes caryophyllene oxide, (E)-nerolidol, β-caryophyllene and monoterpene nerol. The antimicrobial activity of EO was tested against selected ATCC strains of microorganisms, the bacteria Staphylococcus aureus, Pseudomonas aeruginosaand Escherichia coli, and the fungus Candida albicans. The results showed that the investigated EO inhibited the growth of all selected ATCC strains of microorganisms. The best result was obtained against Escherichia colibacteria with MIC value of 1 mgmL-1. The cytotoxicity of EO was measured against the HeLa cell line using the MTT method with IC50of 188.52 μgmL-1.This study has provided scientific baseline data on the therapeutic properties of P. dysenterica. KEYWORDS:Pulicaria dysenterica, essential oil,antimicrobial activity,citotoxicity

Toxicity caused by carbon tetrachloride (CCl4) can lead to serious liver injury. The aim of the study is to investigate the protective effects of oregano oil (Origanum minutiflorum extract oil) against CCl4‐induced liver injury. Two doses of oregano oil were used in the experiment: a low dose (LD; 20 mg/kg) and a high dose (HD; 60 mg/kg) during 2 weeks. CCl4 caused severe liver damage, nucleolus destruction in hepatocytes and cytogenetic changes in the nucleus. Indirectly, CCl4 causes decreased protein synthesis and significantly high creatinine and urea values. Hematological disorders have been recorded, such as decreased RBC and hemoglobin concentration, increased WBC and deformability of the erythrocyte membrane. Both doses of oregano oil had protective effects. Improved protein synthesis and high globulins level, creatinine and urea were found in both groups. Cytogenetic changes in the nucleus of hepatocytes were reduced. A high dose of oregano oil had maximal protective effects for RBC, but a very weak effect on hemoglobin synthesis. Also, WBC and lymphocyte values were low. Origanum stimulates protein synthesis and recovery of hepatocytes after liver injury, reduces the deformability of the erythrocyte membrane. High doses of oregano oil decreased WBC and lymphocytes which may lead to a weakening of the immune response. However, high doses are more effective against severe platelet aggregation than low doses, suggesting an effective treatment against thrombocytosis.

Z. Hodžić, Ivana Nikolić, Aida Crnkić, Nadira Ibrišimović Mehmedinović, A. Kesić, Almir Šestan

In addition to their undeniable importance for industry, natural plant products are widely used in the production of functional food, which, in addition to satisfying nutritional properties, also exhibits certain pharmacological and physiological effects on human health. Chokeberryfruit and products are considered to be excellent sources of polyphenoliccompounds. A large part of polyphenolic compounds from aronia berriesis found in the juice, but the flesh of the berry that remains behind in the juice production process is also richin these bioactive compounds. The aim of the conducted research was to examine the antioxidant potentialof thechokeberry juices from Bosnia and Herzegovina.The antioxidant potentialof chokeberryjuicesin this study wastestedbythe ferric reducing antioxidant power(FRAP)andtotal phenol content (FC). Tests were carried out in other juices for the same parameters in order to compare the obtained values.The research results show that the values of FRAP and total phenols in chokeberry juices vary (20-63 mmol Fe II/L; 850-4930 mgGAE/L), but are significantly higher than in other analyzed juices (7-25 mmol FeII/L; 220–1265 mgGAE/L). By comparing the content of total phenols and FRAP values in pasteurized and unpasteurized chokeberry juice samples, it can be concluded that they are higher in the sample prepared by the pasteurization process. Statistical parameters showthat thelinearcorrelation between thetotal phenols and the FRAP valuesof chokeberry juices (r=0,964; p ˂0,001) and other analysedjuices (r=0,960;p ˂0,001 ) isstatisticallysignificant. KEYWORDS:chokeberry(Aronia melanocarpaL.)juice,antioxidant potential, FC, FRAP

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.

T. Sono, M. Maluleke, N. Ramdas, A. Jelić, Stephen M Campbell, Vanda Marković-Peković, N. Schellack, Santosh Kumar et al.

Introduction: There are concerns with rising rates of antimicrobial resistance (AMR) across countries with appreciable impact on morbidity, mortality and costs. Amongst low- and middle-income countries, a key driver of AMR is the excessive use of antibiotics in ambulatory care, with a critical area being the appreciable selling of antibiotics without a prescription often driven by patient demand and limited knowledge. There is currently conflicting evidence in South Africa regarding this practice. Consequently, there is a need to explore these critical issues amongst patients, especially in more rural areas of South Africa. A pilot study was undertaken to address this. Methods: A two-step descriptive approach was undertaken. This involved two questionnaires amongst patients exiting chain and independent community pharmacies followed by cognitive interviews. Results: Overall, 21 patients were approached for an interview, including 11 for Part 1 of the questionnaire with 3 declining, and 10 for Part 2, with 2 declining. Subsequently 8 patients completed each part of the questionnaire.. On average, it took 2 min 13 s to complete both parts. 3 of the 5 patients being dispensed an antibiotic were dispensed one without a prescription, with all 3 patients exiting from independent pharmacies. Key reasons for self-purchasing included money and convenience. There was mixed knowledge regarding antibiotics and AMR amongst the 8 patients interviewed with Part 2. Overall, there was a satisfactory understanding of the Part 1 questions, although some modifications were suggested. Some participants had difficulty with fully understanding the questions in Part 2, with a number of suggestions made to improve this for the main study. Conclusion: There were concerns with the extent of purchasing antibiotics without a prescription in this pilot study as well as the knowledge of patients regarding antibiotics and AMR. Both areas need addressing and will be explored further in the main study.

I. Ianache, A. Skrzat-Klapaczyńska, D. Jilich, L. Fleischhans, I. Gmizić, J. Ranin, Antonios Papadopoulos, K. Protopapas et al.

Objectives. The purpose of this study was to determine whether fear and prejudice in relation to organ donation and the transplantation of organs may influence the decision to become an organ donor. Materials and Methods. Data were collected through four group interviews using open-ended questions and qualitative content analysis. Forty participants, 16 males and 24 females from seven countries, participated in the focus group interviews. Results. The analysis resulted in three main categories, and nine subcategories. Fears and prejudice caused by tradition and customs, approval of organ donation by family members, perception of the body as a gift from parents, the influence of religious leaders, knowledge about the religious understanding of organ donation, influence of social ambience on respondents, knowledge of the donation process in the healthcare system, including knowing about life after eventual organ donation, were some of predictors in the decision to agree to organ donation. Conclusion. More education on the factors that influence organ donation, more information in schools, health institutions and through the media, as well as more research with the aim of “dispelling” fears and prejudice about organ donation would significantly improve the current situation and result in a larger number of potential organ donors.

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