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Abdurahim Kalajdžić, N. Pojskić, A. Ahmić, Merima Miralem, Lejla Lasić, M. Dzehverovic, Belma Jusić, A. Pilav et al.

Population genetic studies have shown that the Bosnian-Herzegovinian (B&H) population is a part of the European gene pool, but there has been limited information on the genetic structure of ancient B&H populations. This study aimed to determine the frequency and distribution of mitochondrial DNA (mtDNA) haplogroups for a medieval Bosnian population. Thirty-four samples, excavated from medieval necropolises located within the borders of medieval Bosnia, were analyzed. Sequencing of the mtDNA hypervariable segment 1 (HVS1) region and RFLP analysis were performed for haplogroup determination. All 32 samples were identified as haplogroup H, with subhaplogroups H2a and H5 in 30 and 2 samples, respectively. The frequency of the H haplogroup was significantly different between the studied samples and previous studies of contemporary B&H populations, where the H haplogroup frequency was approximately half that of the ancient population studied here. A significant difference in H haplogroup frequency compared with other medieval populations outside of Bosnia was also observed: the ancient B&H population is most similar to ancient Italians. These results provide insight into the mitochondrial landscape of populations that inhabited the territory of present-day Bosnia and Herzegovina in the Middle Ages. Our study reveals that inhabitants of medieval Bosnia carried genetic lineages that exist today in B&H populations, suggesting continuity of mtDNA haplogroups over a long period of time, regardless of various historical demographic events that shaped the genetic structure of the modern B&H population.

M. Meland, M. F. Akšić, O. Frøynes, A. Konjić, Lejla Lasić, N. Pojskić, F. Gaši

In order to best conserve, as well as utilize, traditional apple germplasm in Norway, an apple heritage cultivar collection was established in Ullensvang, western Norway, which aims to become the National Clonal Germplasm Repository. The establishment of the apple heritage cultivar collection was preceded by a molecular study that aimed to genotype a large number of apple accessions maintained in various ex situ sites in western and south-eastern Norway, using a rather small set of eight SSR markers. However limited, the marker set managed to identify synonyms, homonyms, and duplicates within and among the investigated collections. In this study, 171 apple accessions from the Ullensvang apple heritage cultivar collection were genotyped using a set of 20 different SSR markers. Approximately half of the accessions have been previously genotyped using eight SSR markers, enabling an assessment of whether the use of a larger marker set would yield a more accurate characterization. Based on the obtained molecular data, the apple heritage cultivar collection was determined to hold a key part of the overall genetic diversity of the Norwegian apple germplasm. Furthermore, the twelve additional SSR markers were able to differentiate several accessions groups originally thought to be synonyms, as well as to provide a more detailed insight into the genetic structure of this germplasm.

M. Vekić, Belma Kalamujić Stroil, S. Trivunović, N. Pojskić, M. Djukić Stojčić

Abstract Banat Naked Neck is the most important indigenous breed of chickens in Serbia. Marginalized until recently, it is becoming increasingly popular due to its adaptability and good productivity in alternative production systems. However, its history and the current breeding model pose challenges for breed preservation and future improvement. This study aimed to assess the genetic diversity and structure of four subpopulations of Banat Naked Neck from different districts in Serbia (West Backa, North Banat, South Banat and Kolubara) using D-loop mitochondrial DNA sequences and a set of 30 microsatellite markers. Seven haplotypes in the phylogenetic analysis of D-loop mitochondrial DNA suggested maternal origin related to the Indian subcontinent, while haplotype and nucleotide diversity averaged 0.731 ± 0.053 and 0.0067 ± 0.0018, respectively. Microsatellite genotyping showed an average detected number of alleles per locus of 5.129 ± 0.237, while the observed and expected heterozygosity averaged 0.560 ± 0.018 and 0.631 ± 0.014, respectively. Genetic differentiation estimated through FST was 0.051 (p < .001). Two clusters in STRUCTURE analysis showed possible separation of two older subpopulations (South Banat and Kolubara) from the two more recent ones (West Backa and North Banat). This first comprehensive study of genetic diversity serves as the basis for future preservation, use and improvement of the Banat Naked Neck breed.

Silene sendtneri Boiss. (Caryophyllaceae) is the Dinaric endemic plant species with white, decorative and scented flowers. Previous studies on this endemic species were based on morphology and effects on seed germination after the treatment with salicylic acid. However, no molecular genetic studies have been conducted on this species so far. This paper presents preliminary results of the usefulness of microsatellite loci created for cosmopolitan species in assessing the genetic diversity of endemic plant species. A total of 100 specimens were collected from 18 localities in the mountain regions of Treskavica, Igman, Bjelašnica and Ozren in Bosnia and Herzegovina. No S. sendtneri individuals were found at the mountain Trebević. We tested cross-amplification success and a polymorphism level for the set of microsatellite markers (Sil01, Sil03, Sil16, Sil31, Sil35) designed for the cosmopolitan species Silene nutans. In 100 analyzed individuals of S. sendtneri, Sil31 and Sil35 did not amplify, Sil01 was monomorphic and the remaining two loci showed a high level of allelic diversity. Our findings suggest that caution should therefore be exercised in selecting microsatellite markers designed for cosmopolitan plant species in the analyses of endemic species of the same genus since different genetic factors affect the amplification success and polymorphism of the given loci. Attention should be given to the number of detected and effective alleles and their ratio, the success of locus amplification concerning the complete set of markers used, and the ratio of polymorphs to the total number of observed loci.

E. Halilović, A. Ahmić, Abdurahim Kalajdžić, Anel Ismailović, J. Cakar, Lejla Lasić, A. Pilav, M. Dzehverovic et al.

Studies indicate the complex nature of the genetic structure of the European Roma which has been shaped by different effects of their demographic history, while preserving their ancestral Indian origin. The primary aims of this study were to present for the first time the paternal profiles of the Roma from Bosnia and Herzegovina based on the data from Y‐chromosome STR loci, identify the components of non‐Roma paternal gene flow into the Roma, and evaluate the genetic relationships with other European Roma populations.

Background: Identifying risk factors for the occurrence of cardiovascular diseases (CVD) is the first step in developing a prevention strategy. Objective: The aim of the study is to investigate the presence of risk factors and assess the overall risk of developing cardiovascular disease. Methods:.This cross sectional and experimental study involved 203 respondents of both sexes from the working population group aged 40 to 65 years. Results: Hypertension was diagnosed in 154 (75,9%) respondents, hyperglycemia in 177 (87,19%) respondents, dyslipidemia in 182 (89,66%) respondents and Diabetes mellitus type 2 was diagnosed in 53 (26,1%) respondents. Excess body mass was observed in 84 (41,4%) respondents, while obesity was noted in 91 (44,8%) respondents, and smoking habit in 117 (57,6%) respondents. Analyzing the above data using Q3 risk assessment for cardiovascular disease occurrence, it was found that the overall Q3 risk was significantly higher in male respondents, 21,8 (12.4-30.6), while female respondents had a median value of 14,7 (14-22,5), with an apparent statistically significant difference of p=0,001. Conclusion: To establish a strategic prevention program to eliminate the existing risk factors can only be achieved through multidisciplinary planning and the involvement of a multidisciplinary team of experts that will help improve the lifestyle of the individual and the entire community. From this work, we can see that the Q3 risk tool is well suited for risk assessment because it incorporates multidisciplinary parameters, whereas lipid indices can only be useful in cardiovascular disease risk assessment by comparing all indices.

Abstract Interspecific hybridization in the Cyprinidae family has been recorded worldwide, with Abramis brama (bream) and Rutilus rutilus (roach) as one of the often-reported hybridizing pairs. The only account of such an event in Bosnia and Herzegovina has been in Modrac Reservoir. Using morphological and molecular markers, the presence of hybrids was surveyed, the hybridization direction was determined and the hybrid group structure in this ecosystem was evaluated. Our findings confirmed unhindered natural hybridization between roach and bream in Modrac Reservoir. Over 50% of the hybrid specimens were classified as F2 hybrids by the NewHybrids software, while the rest were categorized as pure parental form, making it the first such finding in Europe. The analysis of mitochondrial cytochrome b showed that 90% of hybrid individuals were of bream maternal origin. The hybrid group expressed higher mean values of observed heterozygosity and gene diversity than both parental species. Signs of introgressive hybridization between parental species were detected. The hybrid zone of Modrac Reservoir appears to follow the intermediate or “flat” hybrid model based on the balanced distribution of parental and hybrid genotypes. Further investigation is needed to elucidate the factors that enable the survival and mating success of post-F1 individuals.

This paper provides an overview of the seven possible hand-related traits, with an idea to estimate the statistical phenotypic association between them. The traits observed in this study were: midphalangeal hair, Hitchhiker’s thumb, extensibility of proximal joint of thumb, digital index, nail shape, crooked fifth finger and hand clasping. The mentioned characteristics were observed in 7431 unrelated subjects, with approximately equal gender distribution. For each case, three groups were formed: total, male and female. The subjects were evenly distributed throughout Bosnia and Herzegovina’s various geographical regions, and they properly represent the country’s national and ethnic composition. The Chi-squared test was used to determine the statistical significance of the association between these traits, while Fisher’s exact test was utilized as an extra test to analyze the association between each pair of observed features. Also, Chi-squared test was applied for observing differences in the frequencies of the phenotypic characteristics of the hand between the genders. Following traits were shown to have a statistically significant association: midphalangeal hair-Hitchhiker’s thumb, midphalangeal hair – digital index, midphalangeal hair – nail shape, midphalangeal hair – crooked fifth finger, Hitchhiker’s thumb- extensibility of proximal joint of thumb, digital index – nail shape, digital index -crooked fifth finger, midphalangeal hair – hand clasping, crooked fifth finger – hand clasping. Analysis of differences in the frequencies of the observed phenotypic traits of the hand according to the genders showed statistical significance for D, Dht, Ref and Lf.

Belmina Saric, Nikolina Tomić, Abdurahim Kalajdžić, N. Pojskić, L. Pojskić

Abstract At the end of December 2019, first identified cases of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) started emerging. Ever since the emergence of the first case of infection with SARS-CoV-2 or COVID-19, it became the hottest research topic of numerous studies, in which scientists are trying to understand the path of infection, transmission, replication and viral action, all in order of finding a potential cure or vaccine applying various fundamental principles and methodologies. Using in silico method via AutoDock Vina 1.1.2., we analysed the binding affinity of six selected compounds from grapefruit seed extract (GSE) (narirutin, naringin, naringenin, limonin, ascorbic acid and citric acid) to SARS-CoV-2 main protease Mpro (PDB ID: 6Y84), using acetoside, remdesivir and gallic acid as a positive controls of binding affinity. Results showed highest affinity (rmsd l.b. 0.000; rmsd u.b. 0.000) for narirutin (-10.5), then for naringin (-10.1), acetoside (-10.0), limonin (-9.9), remdesivir (-9.6), naringenin (-8.2), ascorbic acid (-6.7), citric acid (-6.4) and gallic acid (-6.4), all expressed in kcal/mol. Our findings suggest that selected compounds from grapefruit seed extract represent potential inhibitors of SARS-CoV-2 Mpro, but further research is needed as well as preclinical and clinical trials for final confirmation of inhibitory functionality of these compounds.

L. S. J. Medina, Paula B. Paz Sepúlveda, V. Ramallo, C. Sala, Julieta Beltramo, Marisol Schwab, J. Motti, M. R. Santos et al.

abstract:Haplogroup Q originated in Eurasia around 30,000 years ago. It is present in Y-chromosomes from Asia and Europe at rather low frequencies. Since America is undoubtedly one of the continents where this haplogroup is highly represented, it has been defined as one of the founding haplogroups. Its M3 clade has been early described as the most frequent, with pan-American representation. However, it was also possible to find several other haplogroup Q clades at low frequencies. Numerous mutations have been described for haplogroup Q, allowing analysis of its variability and assignment of its geographic origin. We have analyzed 442 samples of unrelated men from Argentina and Paraguay belonging to haplogroup Q; here we report specifically on 27 Q (xM3) lineages. We tested 3 single-nucleotide polymorphisms (SNPs) by amplified product-length polymorphism (APLP) analysis, 3 SNPs for restriction fragment length polymorphism (RFLP) analysis, 15 SNPs by Sanger sequencing, and 17 short tandem repeats (STRs). Our approach allowed us to identify five subhaplogroups. Q-M3 and Q-CTS2730/Z780 are undoubtedly autochthonous lineages and represent the most frequent subhaplogroups, with significant representation in self-defined aboriginal populations, and their autochthonous status has been previously described. The aim of present work was to identify the continental origin of the remaining Q lineages. Thus, we analyzed the STR haplotypes for the samples and compared them with haplotypes described by other authors for the rest of the world. Even when haplogroup Q lineages have been extensively studied in America, some of them could have their origin in post-Columbian human migration from Europe and Middle East.

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.

Interspecific hybridization in the Cyprinidae family has been recorded worldwide, with Abramis brama (bream) and Rutilus rutilus (roach) as one of the often-reported hybridizing pairs. The only account of such an event in Bosnia and Herzegovina has been in Modrac Reservoir. Using morphological and molecular markers, the presence of hybrids was surveyed, the hybridization direction was determined and the hybrid group structure in this ecosystem was evaluated. Our findings confirmed unhindered natural hybridization between roach and bream in Modrac Reservoir. Over 50% of the hybrid specimens were classified as F 2 hybrids by the NewHybrids software, while the rest were categorized as pure parental form, making it the first such finding in Europe. The analysis of mitochondrial cytochrome b showed that 90% of hybrid individuals were of bream maternal origin. The hybrid group expressed higher mean values of observed heterozygosity and gene diversity than both parental species. Signs of introgressive hybridization between parental species were detected. The hybrid zone of Modrac Reservoir appears to follow the intermediate or “flat” hybrid model based on the balanced distribution of parental and hybrid genotypes. Further investigation is needed to elucidate the factors that enable the survival and mating success of post-F 1 individuals.

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.

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