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Ajla Smajlović

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Halogenated boroxine K2[B3O3F4OH] (HB), an inorganic derivative of cyclic anhydride of boronic acid, is patented as a boron-containing compound with potential for the treatment of both benign and malignant skin changes. HB has effectively inhibited the growth of several carcinoma cell lines. Because of the growing interest in autophagy induction as a therapeutic approach in bladder carcinoma (BC), we aimed to assess the effects of HB on metabolic phenotype and autophagy levels in 5637 human bladder carcinoma cells (BC). Cytotoxicity was evaluated using the alamar blue assay, and the degree of autophagy was determined microscopically. Mitochondrial respiration and glycolysis were measured simultaneously. The relative expression of autophagy-related genes BECN1, P62, BCL-2, and DRAM1 was determined by real-time PCR. HB affected cell growth, while starvation significantly increased the level of autophagy in the positive control compared to the basal level of autophagy in the untreated negative control. In HB-treated cultures, the degree of autophagy was higher compared to the basal level, and metabolic phenotypes were altered; both glycolysis and oxidative phosphorylation (OXPHOS) were decreased by HB at 0.2 and 0.4 mg/mL. Gene expression was deregulated towards autophagy induction and expansion. In conclusion, HB disrupted the bioenergetic metabolism and reduced the intracellular survival potential of BC cells. Further molecular studies are needed to confirm these findings and investigate their applicative potential.

Jovan Kulić, Milena Dubravac Tanasković, Milan Kulić, Bojan Joksimović, Ajla Smajlović, Radmila Balaban Đurđević, Nikolina Elez-Burnjaković

This study conducted an initial investigation into the association between ACE gene insertion/deletion (I/D) polymorphisms (rs1799752) and hypertension in the Republic of Srpska, Bosnia and Herzegovina. The study featured two distinct groups, each with 100 subjects, systematically categorized based on hypertension status and gender. DNA was extracted, PCR-amplified, and analyzed by gel electrophoresis. Results revealed a higher prevalence of the DD genotype and the D allele in the hypertensive group, although statistical significance was not observed. The II genotype occurred in 18% of the hypertension group and 21% in the control group. A significant difference was found in allele I frequencies between the two groups (p=0.004), with no gender-related variations in ACE alleles. The limited sample size may have constrained the ability to detect statistically significant differences. The odds ratio for the (DD + ID) genotype compared to II was 1.2110 (95% CI: 0.6006 to 2.4418; p=0.5927), indicating no statistical significance. Furthermore, no significant associations were identified between ACE genotypes and alleles and gender. In summary, this preliminary study suggests a potential trend towards a higher prevalence of the ACE gene D allele and DD genotype in hypertensive individuals. However, due to the small sample size, these associations did not achieve statistical significance in this population. Larger-scale investigations are needed for more definitive insights into the relationship between ACE gene polymorphisms and hypertension.

<p>&nbsp;</p> <p>Regardless of the fascinating progress of humanity, biotechnology and medicine, the outbreak of the global pandemic of the SARS-CoV-2 virus has shown us that we are just as vulnerable as in previous eras when communicable diseases decimated the world&rsquo;s population. But the discoveries made so far at the molecular level allow us to connect knowledge interdisciplinary and find solutions and therapeutic strategies where there seems to be no link. It was the previous coronavirus infections that served as a homologous model for finding the connection between the SARS-CoV-2 virus and autophagy. Autophagy, a conserved universal process of all eukaryotic cells responsible for cell survival under stressful circumstances, has been shown to play a significant role in viral invasions. It contributes to both direct and indirect antiviral responses such the elimination of viruses, the presentation of their antigens, and the reduction of inflammatory responses. The autophagy machinery of host cells can, however, be suppressed, evaded, or used by viruses to their benefit. Therefore, autophagy has an ambiguous role in coronavirus-related infections, especially in COVID-19.</p> <p>&nbsp;</p>

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.

S. Isaković, E. Karahmet, S. Hamidović, J. Tahmaz, Ajla Smajlović

The main goal of this study was to determine the impact of the type of packaging to health safety sausage, and therefore the microbiological compliance with the requirements of the Regulation on microbiological criteria for foodstuffs. The study included the determination of the presence of bacteria of the genus: Salmonella, E.coli, Enterobacteria, Campylobacter, Listeria monocytogenes, and sulfitereducing Clostridiae as well as the total number of bacteria within each individual product. According to estimate the effect of the type of packaging (vacuum, MAP and bulk) to the safety of sausages was performed collecting samples of sausages packed in a vacuum packaging, modified atmosphere packaging of bulk and different manufacturers for each type of packaging. As a result of the study, out of 10 analyzed samples, bacteria from the Enterobacteriaceae family in the range of values from 1.8 to 2.55 CFU/mL., E. coli, in the range of 0, were found by isolation on selective mediafrom 0.51-1.23 CFU/mL. Total bacteria ranged from 2.6-4.02 CFU/mL. Yeasts and molds in the samples tested were between 2.22 – 3.84 CFU/mL. Microbiological tests did not reveal any bacteria from the following groups: Salmonella spp., Listeria and sulphite-reducing Clostridia. It was found that all samples correspond to the Regulation on microbiological safety.

Autophagy is a dynamic process, conserved in all eukaryotes. It is responsible for the degradation of cytoplasmic content. Autophagy is crucial in cell survival and cell death. It plays a significant role in the cell response to stress, nutrient deficiencies, embryonic development, tumor suppression, response to pathogens and aging. The process of autophagy is also involved in the pathology of human diseases, such as cancer, diabetes, cardiomyopathy, and neurodegenerative diseases such as Alzheimer's and Parkinson's disease. Autophagy is a mechanism that involves degradation of cells, proteins, damaged organelles and pathogens through the lysosomal mechanisms, thus autophagy supports cell survival during starvation, hypoxia and metabolic stress. However, if extensive and/or excessive, autophagy can promote apoptosis (type I) or function as an alternative cell-death pathway, called autophagic cell death (type II). Autophagy can either promote cancer cell death, or serve as a survival mechanism against apoptosis or necrosis induced by various anticancer treatments. Given the contradictory role of autophagy during tumor initiation and progression, the use of autophagy in therapy depends on the context and must be approached individually

Amar Pecikoza, Lejla Lasić, G. Radosavljević, A. Ahatović, Ajla Smajlović, A. Durmić-Pašić, L. Pojskić, N. Pojskić

Apart from its physiological role in the cellular oxidation of ethanol interesting feature of the ADH1B gene locus is its characteristic geographical distribution in which certain variants of ADH1B peak in different parts of the world.  Therefore, ADH1B rs2066701 polymorphism is exploited as a genetic marker in tracing of the evolutionary processes and human migrations in the past. Taking into consideration the complexity of population genetic structure and several migrations in the history of the Balkan populations, including Bosnian and Herzegovinian, this study aimed to estimate the frequency of ADH1B rs2066701 polymorphism in the population of Bosnia and Herzegovina. The total of 101 randomly sampled individuals was genotyped for rs2066701 polymorphism in ADH1B gene using PCR-RFLP method. The obtained frequencies were used to calculate heterozygosity, fixation indices and Hardy-Weinberg equilibrium. Observed population-structure parameters were compared with other population values available in ALFRED database. Dimensional relations between the investigated populations were visualised with the NM-MDS (non metric multidimensional scaling) analysis using PAST. The minor allele frequency for rs2066701 was 0,257. Inter-population analysis including other European and non-European populations from the ALFRED database proved the above-mentioned European genetic background of the B&H population.

Amar Pecikoza, Lejla Lasić, G. Radosavljević, A. Ahatović, Ajla Smajlović, A. Durmić-Pašić, L. Pojskić, N. Pojskić

Apart from its physiological role in the cellular oxidation of ethanol an interesting feature of ADH1B gene is its characteristic geographical distribution where certain variants of ADH1B peak in different parts of the world. Therefore, ADH1B rs2066701 polymorphism is used as a genetic marker in tracing the evolutionary processes and human migrations over time. Taking into consideration the complexity of population genetic structure and a number of migration events in the history of the Balkan populations this study aimed to estimate the frequency of ADH1B rs2066701 polymorphism in the population of Bosnia and Herzegovina. The total of 101 randomly sampled individuals were genotyped for rs2066701 polymorphism in ADH1B gene using PCR-RFLP method. The obtained frequencies were used to calculate heterozygosity, fixation indices and HardyWeinberg equilibrium. The observed population-structure parameters were compared with other population values available in ALFRED database. Dimensional relations between the investigated populations were visualised with the NM-MDS (non metric multidimensional scaling) analysis using PAST. The minor allele frequency for rs2066701 was 0,257. Inter-population analysis including other European and non-European populations from ALFRED database proved the above-mentioned European genetic background of the B&H population.

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