This study was designed on the analysis of the mtDNA polymorphisms in three ethnic populations of Tuzla Canton of Bosnia and Herzegovina (Bosniaks, Croats and Serbs). The main aim of this study was to analyze the influences of the maternal gene flow on the genetic profile of the analyzed populations. The analysis of mtDNA variation based on relevant restriction fragment length polymorphisms (RFLP) in combination with HVSI variations of the control region (for detection of subhaplogroups of the haplogroup U) enabled the identification of the typical of the Western-Eurasian haplogroups (H, I, J, T, W, U, HV, HVO, K, V, and X), African/Near East lineages N1a and Asian haplogroup M. Our results suggest that mitochondrial gene pool of the three main ethnic groups of Tuzla region was shaped by influences of early and late migration routes which marked the settlement process of the Balkans. The effects of different migration directions are illustrated by the distribution of important indicators of the Late Glacial expansion (U5a), postglacial re-colonisation of Europe from glacial refuges of southwestern European (H, V, U5b1), central-eastern European Plain (U4), Italian Peninsula (U5b3) and neolithic expansion (U3, N1a, J and T). Our data can indicate a common genetic history, origin, as well as a similar contribution of the parental and maternal gene flow on genetic structure of the three main ethnic populations of modern Bosnia and Herzegovina.
Of the four species of the genus Satureja (Lamiaceae) that are recognized in Bosnia and Herzegovina, S. subspicata has the the widest distribution. It is taxonomically challenging species of geographically limited distribution and little data on its genetic diversity throughout its range is available. We sampled six geographically distinct populations from Bosnia and Herzegovina and applied nrDNA (ITS1, ITS2), chloroplast markers (matK and trnL) and AFLP to examine genetic diversity of S. subspicata in the center of its distribution range and to explore the possibility of establishing the species DNA barcode. AFLP analysis showed large genetic differentiation among populations as well as moderate correlation between genetic distance among populations and geographic distance among locations. MatK has not proven useful in distinguishing S. subspicata from sympatric species. However, nrDNA sequences provided necessary resolution power, with ITS2 being more informative. Estimates of evolutionary divergence between nrDNA sequences obtained in our research and homologous sequences of sympatric Satureja deposited in the GenBank reveal closer relationship between geographically proximate populations of different species and slight divergence within S. subspicata sequences pool. This outcome highlights the importance of considering overall genetic diversity across the distribution range of a species when assigning DNA barcode.
In the present study modern technology of DNA extraction and automatic genotyping was applied in Bosnian and Herzegovinian autochthonous horse breed by using 17-Plex horse genotyping kit. The study was aimed at investigating usefulness of the 17-plex STR Kit for Bosnian mountain horse genotyping and establishing highly useful microsatellite markers system for genetic diversity studies in Bosnian mountain horse breed. Genomic DNA was extracted from whole blood collected from 22 unrelated Bosnian mountain horse specimens. A total of 95 alleles were detected. Average number of detected alleles per locus was 5.588, varying from 3 (HTG7) to 10 (ASB17). Average effective number of alleles was 3.603, fluctuating from 1.789 (HMS7) to 5.728 (HMS2). The observed heterozygosity ranged from 0.136 (HMS3) to 0.909 (ASB2) with a mean of 0.631. The results indicate that the studied population originates from the appropriate number of parent generations. The mean expected heterozygosity was 0.690, varying from 0.441 (HMS7) to 0.853 (ASB17) indicating high genetic variability within Bosnian mountain horse population. The PIC values ranged from 0.409 (HMS7) to 0.837 (ASB17) with a mean of 0.643, suggesting that 94.12% markers were quite informative in terms of their suitability for genetic diversity studies .The most polymorphic locus was HMS2 and the least polymorphic locus was HMS7. The inbreeding coefficient ranged from -0.030 (HMS7) to 0.807 (HMS3) with a mean of 0.077. Inbreeding coefficient values indicated no shortage of heterozygotes in Bosnian mountain horses. Deviation from Hardy-Weinberg equilibrium (p<0,05) was found in three loci (HTG10, HMS3 and ASB17). The applied set of 17 microsatellite markers proved to be sufficiently specific for use in genotyping of Bosnian mountain horse. Considering the values of HO, HEand PIC over 0.6, five microsatellite markers system (HTG4, AHT4, AHT5, ASB2, HMS2) is considered to be highly useful for genetic diversity studies in Bosnian mountain horse breed.
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 This article presents a new approach to detect coiled coil and leucine zipper (L-Zip) motifs in protein sequences. The approach is based on protein scale calculation and sequence analysis. For this purpose, the wavelet-based local extrema extraction is employed, and window-based variations of local extrema afterward. This, in turn, provided a way to distinguish coiled coil subsequences and potential L-Zip motifs. The approach is validated on carefully chosen protein sequences that return inconclusive results within known frameworks for L-Zip detection, for example, 2ZIP. The results show that this new approach represents an improvement over previously presented approaches.
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
Conventional screening and diagnostic procedures in prostate complaints rely on PSA (Prostate Specific Antigen) concentration which is not specific for prostate cancer and frequently leads to unnecessary invasive procedures in order to exclude malignant disease. It is estimated that approximately 50% of persons who underwent tissue biopsy did so based on false positive PSA value. Therefore a proper and timely differential diagnosis of malignant disease using non-invasive techniques remains one of the biggest challenges in medicine. Urine is the invaluable source of biological information contained in small molecules i.e. RNA that is easily accessible and detectable using molecular genetics techniques. We describe economical and fast method for relative expression analysis applicable to any target gene using urine as a sample. Efficient non-invasive method for identification of malignant or high risk cases prove useful in reduction of patient distress during the diagnostic procedure and significantly reduce healthcare costs.
Population differentiation based on genetic diversity was subject of many previous scientific studies. Consequently, various methods were suggested. The most widely used method was fixation index FST, as a part of FIS, FIT and FST parameters which were proposed by Wright (1943, 1951, 1965). The main objective is to hierarchically estimate genetic variation in populations. Nei (1973, 1987) suggested GST as more appropriate methods, with θ (Cockerham 1969, 1973; Weir et Cockerham 1984), and ΦST (Excoffier et al. 1992) introduced later on as more adequate methods for molecular markers. Wright’s FST has range between 0 and 1 where 0 indicates absence of differentiation, while 1 shows absolute divergence with no shared alleles. This method helps to quantify and compare level of genetic differentiation among populations. Since, in practice, when multialleles loci are applied, Fst value of 1 is almost never observed for fixation indices (Wright 1978; Hedrick 1999; Jost 2008). This fact reduces application of fixation indices when highly polymorphic markers (e.g microsatellites) are used (Hedrick 1999). However, certain literature suggests that Nei's GST and Wier and Cockerham's θ are flawed in the sense that 1 does not represent maximal differentiation. Arguing about practical applicability of standard genetic differentiation methods, Jost (2008) suggested allelic diversity (∆) to be base for measuring the genetic differentiation Dest as indicator of divergence (D). Jost considers that this approach corrects sampling bias, does not suffer the flaws of F-statistics and, being related to diversity, is more adequate. Nilsryman and Olofleimar (2009) concluded in their study that Dest suffers the same problems as other measures, and that GST is still more appropriate method.
The expert reports state that Bosnia and Herzegovina, despite the presence of diverse and valuable natural resources, lacks systematic, coordinated and harmonized pipeline for biomonitoring. Successful solutions to serious problems regarding environmental protection, management and research rely on the efficient use of exhaustive and unfailing information on the nature around us. However, more often than not, transitional and developing countries lack any centralized, nationally funded databases that could be used as dependable source of information in decision making process. University of Sarajevo-Institute for Genetic Engineering and Biotechnology (INGEB) developed the Regional Biodiversity Database – REBIDA with the aim to collate all known biological data on wild and domesticated natural resources of Bosnia and Herzegovina. This internet-based database represents a comprehensive, searchable and open access platform for science community, academia, governmental and non-governmental stakeholders and general public. Besides its scientific value, REBIDA will serve as an educational tool for discovering the diversity and importance of natural resources, with special emphasis on indigenous and endemic flora, fungia and fauna from the Balkans. It is the only such database in the country, consisting of three functionally connected segments: tissue database, DNA database and digital genetic database on plant, animal and human samples. To complement REBIDA, a mobile application called REBIDA SCANNER was also developed. It will be free to download for IOS and Android platforms and will enable professionals, nature enthusiasts and any other interested parties to contribute to REBIDA through data collection, field sampling and documentation of B&H wild life.
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