ABSTRACT Six blood groups (Rh, MN, Duffy, Kidd, Kell, and Lutheran) were investigated among three major ethnic groups (Bosniaks, Bosnian Croats, and Bosnian Serbs), as well as 10 regional subpopulations across Bosnia and Herzegovina (B&H): Krajina; Posavina; northeastern, eastern, middle, and central Bosnia; Sarajevo region; eastern, central, and western Herzegovina. This is the first study that introduces the molecular genetic typing of five blood groups within the B&H population, with the exception of the RhD blood group. The sample consisted of 450 buccal swabs from unrelated individuals. Five blood group systems (RhD, RhC, RhE, Kidd, MN) were genotyped by PCR with sequence specific primers, while three blood group systems (Kell, Duffy, Lutheran) were genotyped by the PCR-restriction-fragment-length polymorphism method. Minor variation of genetic diversity was observed within the three major B&H ethnic groups, as well as within the 10 subpopulations stratified according to geographical criteria. No genetic differentiation among ethnic groups was noticed. These results are in agreement with the results of previous studies based on different molecular genetics markers, which indicate that the three B&H ethnic groups belong to the same gene pool. A similar level of genetic variance was observed within regional subpopulations, with no significant genetic differentiation among them. Comparison of intrapopulation genetic diversity of the B&H population with other European and non-European populations, based on three loci (RHD, MN, and KEL), clearly show that the level of genetic diversity of the B&H population is within the European range.

Six blood groups (Rh, MN, Duffy, Kidd, Kell, and Lutheran) were investigated among three major ethnic groups (Bosniaks, Bosnian Croats, and Bosnian Serbs), as well as 10 regional subpopulations across Bosnia and Herzegovina (B&H): Krajina; Posavina; northeastern, eastern, middle, and central Bosnia; Sarajevo region; eastern, central, and western Herzegovina. This is the first study that introduces the molecular genetic typing of five blood groups within the B&H population, with the exception of the RhD blood group. The sample consisted of 450 buccal swabs from unrelated individuals. Five blood group systems (RhD, RhC, RhE, Kidd, MN) were genotyped by PCR with sequence specific primers, while three blood group systems (Kell, Duffy, Lutheran) were genotyped by the PCR-restriction-fragment-length polymorphism method. Minor variation of genetic diversity was observed within the three major B&H ethnic groups, as well as within the 10 subpopulations stratified according to geographical criteria. No genetic differentiation among ethnic groups was noticed. These results are in agreement with the results of previous studies based on different molecular genetics markers, which indicate that the three B&H ethnic groups belong to the same gene pool. A similar level of genetic variance was observed within regional subpopulations, with no significant genetic differentiation among them. Comparison of intrapopulation genetic diversity of the B&H population with other European and non-European populations, based on three loci (RHD, MN, and KEL), clearly show that the level of genetic diversity of the B&H population is within the European range.

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.

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

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.

Abstract Although prostate cancer accounts for the highest number of newly diagnosed cases of cancer in men, it represents a specific diagnostic challenge in modern oncology. The standard diagnosis of prostatic carcinoma begins with the screening of serum concentrations of PSA (Prostate Specific Antigen). If the concentration of serum PSA levels is above 4 ng/mL, the patient is further referred to a digital rectal examination in order to determine an increase in prostate volume. In cases where enlargement of the prostate is observed, the next step is biopsy of prostate tissue. This physically painful and invasive approach to confirm the diagnosis is often unnecessary because, in many cases, the patohistologic analysis determines diagnosis of benign prostatic hyperplasia, and not a tumor. In this study, we investigated the possibilities of detection and measurement of the relative level of gene expression of the KLK3 (Kallikrein-related peptidase 3), PCA3 (Prostate Cancer Gene 3) and TEMPRSS: ERG (Transmembrane protease serine2 and in-ETS erythroblostosis virus E26 oncogene homolog) genes from the urine samples of patients with prostatic diseases and healthy controls. Urine was the sample of choice because it is taken in a non-invasive manner, and could potentially serve to make better selection to biopsy. One of the selected genes (KLK3) differed significantly in the samples of various pathological conditions of the prostate, and therefore we consider that its further investigation is reasonable.

Objectives: The global burden of the current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causing the corona virus disease-19 (COVID-19) is enormous No definitive treatment and prophylactic guidelines for COVID-19 currently exist except for physical distancing and aerial barriers between individuals This work explored the natural compound-binding efficiency of SARS-CoV-2 proteins essential for host cell interaction and infection Methods: The binding activity of artemisinin to SARS-CoV-2 spike glycoprotein (Protein Data Bank (PDB) ID: 6VYB), SARS-CoV-2 main protease (3C-like main protease (3CLpro);PDB ID: 6Y84) and SARS-CoV-2 papain-like protease (PLpro;PDB ID: 6W9C), were tested using in silico methods Moreover, chloroquine and hesperidin were used as the positive control of binding affinity and proven therapeutic effect, respectively Results: The highest affinities for binding to all tested SARS-CoV-2 proteins are observed for hesperidin (-5 8,-10 0, and -8 1 kcal/mol), then for artemisinin (-4 8,-8 3, and -6 0 kcal/mol), and the lowest for chloroquine (-4 1,-8 2, and -4 8 kcal/mol) Artemisinin, hesperidin, and chloroquine had similar positioning toward targeted proteins at specific sites when these interactions were visualized Conclusion: This study shows that artemisinin has the potential to bind and inhibit the SARS-CoV-2 spike protein, the 3CLpro main protease, and PLpro proteinase similar to hesperidin and chloroquine that have been proven as antivirals in previous preclinical and clinical studies

Apoptosis, as a well-studied process of a programmed cell death, is essential for the maintenance of cell homeostasis and integrity of organisms. This process occurs normally during development and aging and it is a balance of the sustainability of the tissue cell population. In addition, apoptosis also occurs as a defensive mechanism such as an immune response or after cell damage as a consequence of a pathological condition or the action of harmful agents. Apoptotic activation tends to be less responsive with aging, causing accumulation of non-functional cells and pathological changes such are degenerative diseases or tumor transformation. This overview aims to provide summarized facts about different approaches of apoptosis research, targeting and regulation in tumors especially in leukemic cells as a way of pharmacological manipulation with a potential therapeutic benefit.

MODY (maturity-onset diabetes of the young) is an autosomal dominant form of diabetes that is usually manifested before the 25-year of life. This type of diabetes is caused by defects in the primary insulin secretion. There are several types of MODY, which are monogenic diseases, where mutations in a single gene are responsible for a particular type of MODY. Currently, there are eleven types of MODY, from which the most common types are MODY 2 and MODY 3 (with mutations on GCK and HNF1A genes, respectively). We identified very rare MODY 7 type of diabetes in three family members by MLPA analysis.

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.

Abstract Plant bioflavonoids are widely present in the human diet and have various protective properties. In this study, we have demonstrated the capacity of delphinidin and luteolin to increase human telomerase reverse transcriptase (hTERT) expression level and act as protective agents against halogenated boroxine-induced genotoxic damage. Halogenated boroxine K2(B3O3F4OH) (HB), is a novel compound with potential for the treatment of both benign and malignant skin changes. In vivo and in vitro studies have confirmed the inhibitory effects of HB on carcinoma cell proliferation and cell cycle progression as well as enzyme inhibition. However, minor genotoxic effects of HB are registered in higher applied concentrations, but those can be suppressed by in vitro addition of delphinidin and luteolin in appropriate concentrations. Fresh peripheral blood samples were cultivated for 72 h followed by independent and concomitant treatments of HB with luteolin or delphinidin. We analyzed the differences in relative hTERT expression between series of treatments compared with controls, which were based on normalized ratios with housekeeping genes. The obtained results have shown that selected bioflavonoids induce upregulation of hTERT that may contribute to the repair of genotoxic damage in vitro.

RELIABILITY OF URINE AS SOURCE FOR BIOLOGICAL INFORMATION FOR RISK ESTIMATION FOR PROSTATE MALIGNANCY

Incidence of breast cancer ranges from 27 per 100,000 in Middle Africa and Eastern Asia to 92 per 100,000 in Northern America. It is the fifth most common cause of death from cancer in women, with an estimated 522,000 deaths per year (6.4% of the total). Autosomal dominant inheritance of these cancers is characterized by transmission of cancer predisposition from generation to generation, with around 5-10% of all breast cancers being associated with inherited mutations in BRCA1, BRCA2 and other genes.  Breast and ovarian cancers are strongly associated with BRCA1 and BRCA2 mutations. In this study, we genotyped BRCA1 gene for large genomic rearrangements in breast and ovarian cancer patients from Bosnia and Herzegovina, with aim to assess frequency of large BRCA1 mutations (exon deletions/duplications) in this group. We collected 59 breast cancer samples, as well as other data concerning patients’ histopathological parameters of tumor, like age at diagnosis, cancer type, TNM class, cancer grade, as well as estrogen, progesterone and Her2/neu expression. Following DNA extraction from breast cancer samples (tissue after biopsy), BRCA1 mutations were identified by Multiplex Ligase - Dependent Probe Amplification (MLPA) analysis. Biostatistical analyses were conducted using MedCalc v.9.2.0.0 software. In all statistical tests p<0.05 was considered significant. Mean age at diagnosis was 54±1.75 (range 17 – 80). BRCA1 genomic rearrangements were found in 22% of breast and ovarian cancer patients. Statistically significant associations and correlations were found between BRCA1 genomic rearrangements and cancer type, estrogen, progesterone and Her2/neu expression, but not cancer grade, size, invasiveness or patients’ age