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Abstract Anti-proliferative effects of halogenated boroxine – K2(B3O3F4OH) (HB) – have been confirmed in multiple cancer cell lines, including melanoma, but the exact mechanism of action is still unknown. This study aimed to determine its cytotoxic effects on human Caucasian melanoma (GR-M) cell growth in vitro as well as on the expression of cell death-related genes BCL-2, BECN1, DRAM1, and SQSTM1. GR-M and peripheral blood mononuclear (PBM) cells were treated with different HB concentrations and their growth inhibition and relative gene expression profiles were determined using the Alamar blue assay and real-time PCR. HB significantly inhibited cell growth of both GR-M and PBM cells but was even more effective in GR-M melanoma cells, as significant inhibition occurred at a lower HB concentration of 0.2 mg/mL. GR-M BCL-2 expression was significantly downregulated (P=0.001) at HB concentration of 0.4 mg/mL, which suggests that HB is a potent tumour growth inhibitor. At the same time, it upregulated BCL-2 expression in normal (PBM) cells, probably by activating protective mechanisms against induced cytotoxicity. In addition, all but the lowest HB concentrations significantly upregulated SQSTM1 (P=0.001) in GR-M cells. Upregulated BECN1 expression suggests early activation of autophagy at the lowest HB concentration in SQSTM1 cells and at all HB concentrations in PBM cells. Our findings clearly show HB-associated cell death and, along with previous cytotoxicity studies, reveal its promising anti-tumour potential.

The aim of the paper is to present the results of the VNTR gene polymorphism genetic variants molecular typing for endothelial nitric oxide synthase (eNOS) in human population of Tuzla Canton. Based on the analysis of the distribution of eNOS gene genotypes in the total sample of respondents, the highest frequency was recorded for the (b/b) genotype, which was 73.0%. For the heterozygous (a/b) genotype of the eNOS gene, a frequency of 24.0% was determined, and a frequency of 3.0% was recorded for the (a/a) genotype. The research resulted in a database of local and global significance, namely, the incorporation of these data into the existing regional and European genetic database.

Apoptosis induction is a promising approach in targeting tumor cells. As halogenated boroxine (HB) shows antitumor activity, but its mechanism of action in hematological tumors remains unclear, in this study, we aimed to analyze apoptosis triggering in normal and UT‐7 leukemia cells by HB. Methods for assessing cell viability and cytotoxicity, apoptosis detection, relative expression of 84 apoptosis‐associated genes and BCL‐2, and functional analysis were applied. Pronounced HB activities in inhibition of cell viability, cytotoxicity, and apoptosis induction with measurable differences between tumor and normal cells were found. HB modulated the expression of 21 genes, predominantly downregulated the antiapoptotic genes in leukemia. The functional association revealed HB's impact on inhibition of NF‐κB signaling pathway. BCL‐2 expression decreasing was found only in UT‐7 leukemia. This study identified HB as an apoptosis inducer affecting leukemia but not normal cells considering mechanisms of selective activity that may be a great advantage of HB applications.

Irina Milovač, V. Vidović, J. Ramic, N. Lojo-Kadrić, M. Hadzic, Z. Mavija, Stojko Vidović, L. Pojskić

Background/Aim: Irritable bowel syndrome (IBS) belongs to the gastrointestinal disorders characterised by abdominal discomfort and pain, altered constipation, diarrhoea and stomach distension. The aim was to assess relationship between the selected genetic polymorphisms with IBS, their combined genotype effect as well as to assess a difference in the distribution of allele and genotype frequencies of selected loci between case and control group. Methods: This was a prospective study which included 29 participants, 20 individuals diagnosed with IBS based on Rome III criteria and 9 healthy individuals. The study analysed the selected genetic polymorphisms as possible risk factors for IBS according to the model of the case-control study. Genotyping was performed for FKBP5, DRD2 and DAT polymorphisms qualified as risk factors for IBS in previous researches. Results: The results revealed a significant association between DAT polymorphism with IBS, both, at the allelic level (p = 0.006) and genotype level (p = 0.031). Individuals with 434 allelic variant in the genotype have six time higher probability for developing IBS, in comparison to the individuals without this allelic variant. The statistical association between other analysed polymorphism and IBS was not reached. The analysis of combined effects of selected polymorphisms revealed no association with IBS, except FKBP5 and DAT which result was at the level of statistical significance (p = 0.05). Conclusion: Further analysis which would include DAT polymorphism with larger sample size, as well as other genes involved in dopamine neurotransmitter system would be of great interest to define closer conclusion of IBS aetiology.

CAT gene encodes catalase, a key antioxidant enzyme in the body against oxidative stress. This enzyme plays an important role in the molecular mechanisms of inflammation, apoptosis, mutagenesis and tumorigenesis. Anti-oxidant L-carnitine is used in food supplementation, medical co-treatment and bodyweight regulation. We aimed to investigate molecular basis of L-carnitine commercial preparations supplementation in reducing oxidative stress with customized CAT gene assay in vitro. Human lymphocytes cell culture was established using standard procedure and treated with range of concentrations of L-carnitine in two preparations. We tested two preparations: 500 mg tablets of L-carnitine and liquid L-carnitine with vitamin B6. L-carnitine significantly reduced the expression of CAT gene in cultured lymphocytes at concentrations of 50 μmol/l and 250 μmol/l compared to negative control, (p = 0,001; p = 0,001; respectively). The L-carnitine liquid supplement with vitamin B6 also reduced the transcription of CAT gene at concentrations of 50 μmol/l and 250 μmol/l as compared to the negative control (p = 0,018; p = 0,006; respectively). Selected L-carnitine preparations modulated the transcriptional activity of the antioxidant enzyme gene in human lymphocyte culture, indicating its possible effects in inhibition of pro-inflammatory processes that involve catalase activity.

Nina Bosankić, Selvira Draganović, J. Ramic, A. Haque

This study explored how women breast cancer survivors who underwent radical mastectomy experienced stress and adversity and managed their diagnosis and treatment. This study is based on semi-structured and in-depth interviews with a convenience sample of 22 participants. Qualitative analysis and discussion groups were conducted in the participant’s homes over 18 months. Thematic analysis resulted in four overarching categories that illustrated how being a woman was challenged and restructured from the participants’ personal experiences. The participants’ coping strategies were primarily reflected in their spirituality, optimism, the embrace of healthy lifestyles, and pink ribbon activism.

Selvira Draganović, Nina Bosankić, J. Ramic

Abstract. Introduction: Prenatal, perinatal and postnatal period result in series of psychological, physical, relational and emotional changes and adjustments while during the pandemic pregnant women and mothers of young children must also cope with the fear of themselves, their fetuses, or children being infected. Aim: The aim of study was to explore the lived experiences of pregnant women and mothers living in Bosnia and Herzegovina during the COVID-19 outbreak. Methods: Semi-structured in-depth interviews with 30 Bosnian women, 15 of whom were pregnant and 15 who are mothers that gave birth during the COVID-19 pandemic were conducted. Participants were recruited through two local women’s associations. The data was analyzed utilized inductively using an interpretative phenomenological analysis. Results: The study results indicate that both pregnant women and mothers alike described negative and positive feelings about pregnancy and motherhood during the COVID-19 pandemic such as fear and hope. The lived experiences of anxiety and adaptation through two core concepts: trapped in the fear of the unknown and adapting and embracing uncertainty. Discussion: The main themes that emerged from the interviews reveal grounded fear but also adaptability. Even though people have amazing abilities to adapt to adverse life conditions, as women in our study demonstrate, many aspects of the pandemic’s impacts on vulnerable populations are still unexplored. Tailor-made public health strategies such as an online counseling platform should be created to accommodate specific needs and issues of this population.

J. Ramic, Irina Milovač, Z. Mavija, N. Lojo-Kadrić, M. Hadzic, Stojko Vidović, B. Niesler, N. Dovrolis et al.

Irritable bowel syndrome (IBS) is a functional gut brain gastrointestinal (GI) disorder, typically accompanied by constipation or diarrhea, usually without any organic evidence. The prevalence of IBS is rather high of about 10-15% (10, 1 % according to Rome III and 4, 1% according to Rome IV, Enck P. et al 2016, Sperber A.D. et al 2020, Black C.J. et al 2020) in the working population. Quality of life in patients with IBS is reduced and therefore a major obstacle to the normal physical and social wellbeing. In intensified clinical research worldwide new pathogenic mechanisms of IBS are suggested, including intestinal dysbiois one of the critical contributing factors to onset or further development of IBS. Intestinal microbiome represents a real ecosystem of microorganisms and human GI tract lining cells. The diversity and composition of the GI microbiome may differ significantly inter- and intra-individually, depending on sex, age or physiological conditions (pregnancy, disease, etc). Intestinal microbiome composition frequently changes in association with IBS symptoms, and the purpose of this study was to investigate if there is a clear relationship in microbial composition and relative abundance of microbial taxa in feces of persons diagnosed with IBS. Fecal microbiota profiling was done in a group of nine clinically confirmed IBS patients and 6 corresponding healthy controls, based on species specific 16s RNA gene. No statistically significant differences in Alpha and Beta diversity indices were found.

Luteolin and delphinidin are the flavonoids with known protective roles. They inhibit genotoxic effects induced by halogenated boroxine (HB) in vitro. Statistically significant decrease in the frequency of micronuclei and nuclear buds and suppression of the occurrence of aberrant cells were observed before, but mechanism of its anti-genotoxic activity is still not clear. In our experiment we aimed to quantify HB effects on the relative expression of CAT (catalase) gene and explore antioxidative effects of luteolin and delphinidin via restoration of CAT gene activity. Cell cultures from peripheral blood lymphocytes of five healthy donors were established and treated with independent and concomitant treatments of HB with luteolin or delphinidin. Total RNA was isolated from harvested cells and reverse-transcribed. SYBR based Real-Time PCR amplification method was used. Analysis of results included normalization of ratio of target (CAT) and housekeeping (GAPDH) gene and statistical analysis (REST®). Luteolin itself lead to downregulation of relative CAT gene expression as well as HB. But simultaneous treatment of HB and bioflavonoids lead to upregulation. Delphinidin as independent treatment and as simultaneous treatment caused upregulation of relative CAT gene expression. Obtained results may indicate protective role of delphinidin and luteolin to oxidative damage caused by HB, and also that new approaches to the treatment applications of HB should include bioflavonoids and monitoring corresponding antioxidant system. Our findings indicate that there is a quantifiable effect of luteolin and delphinidine on antioxidant genes which could be used in exact monitoring of oxidative stress related events.

Nikolina Tomić, Adi Osmanbegovic, A. Mujala, D. Prtvar, M. Hadzic, J. Ramic, N. Lojo-Kadrić, N. Pojskić et al.

Background Visceral Sensitivity Index (VSI) questionnaire measures gastrointestinal specific anxiety a mediator of the relationship between general psychological distress measures and gastrointestinal symptom severity. Studies have shown that angiotensin converting enzyme (ACE) may be required for sympatoadrenal activation during stress. The aim of our study was to explore the relationship of ACE gene polymorphisms with the scores for self-reported visceral hypersensitivity in the sample of student population exposed to psychological distress. Methods A blood sample was taken from ninety students during exam period. DNA was isolated and genotyping of ACE polymorphism (rs1799752) was performed using PCR method. The PCR products were analysed on a 2% agarose gel. All respondents completed the VSI questionnaire and based on the scores were stratified into two comparison groups. Allele and genotype association was tested using Fisher’s Exact Test in WINPEPI. Results Respondents with total score of up to 65 were classified in the first group and with values over 65 in the second group. Increased frequencies of D allele and DD genotype were observed in the subgroup of students with higher VSI score. Conclusions Obtained results revealed statistically significant association of allele D and DD genotype with increased VSI score. Our results indicate that further genetic and genome studies of regulation of brain-gut axis and visceral hypersensitivity could be helpful in clinical interpretation of their impact on functional gastrointestinal disorders (FGID) symptoms and on development of some other acute and chronic stress related conditions in youth population.

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