The region of Western Balkans has been inhabited since the Paleolithic era and was the route of the spread of farming from the Middle East to Europe during the Neolithic era. In the present study, Y-STR data from European populations have been used to construct median-joining networks. The study was performed using Whit Athey’s Haplogroup Predictor, Y Utility and Network 4 software packages to predict Y haplogroups, construct networks, perform clustering of closely related Y chromosomes and calculate time estimates between individual nodes. The results of the study imply that geographically close populations cluster together at both Balkan and European levels. It was observed that an elevated number of study populations and individual haplogroups increases the possibility that individuals of different ethnic background cluster within the same or neighboring clades of network. Subsequent time estimates, performed based on the mutation frequency between the ancestral node and its descendant nodes, revealed that I2a haplogroup within the Western Balkan region has the most compact clustering (age, estimated at 3109 years), followed by Hg E1b1b which has the second most compact clustering (4896 years). The obtained results are nonetheless in accordance with previously published research investigating the frequency of Y haplogroups based on Y-SNP variant frequencies, indicating that Western Balkan countries are mainly represented by I2a subclade (average for six countries 32.3%), followed by E1b1b and R1a (average for six countries of 21.5% and 17%, respectively).

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.

Since the introduction of the term low copy number DNA, also referred as low template DNA, touch DNA or trace DNA analysis, it has quickly become focal point of forensic DNA testing as well as other DNA based studies. Low template DNA (ltDNA) samples can be described as the samples which involve single source samples with template DNA in concentrations below 100 picograms (pg). Due to sensitivity of ltDNA samples to contamination, it is of great importance to optimize performance of the multiplex STR systems and existing protocols to increase chance of successful analysis. The main objective of this study was analysis of 20 challenging samples (skeletal remains, cigarette buts, chewing gum, poorly collected buccal swabs etc.) mostly low template DNA samples, preliminarily profiled by PowerPlex® 16 multiplex STR systems and additionally processed with new generation multiplex STR kit PowerPlex® Fusion. Sample isolation was done using a standard phenol-chloroform method for bone samples and DNeasy® Blood and Tissue Kit for other forensic samples. PowerPlex® 16 (PP16), multiplex STR system and PowerPlex® Fusion (PP Fusion) were used for co-amplification of 15 and 24 autosomal STR loci respectively. Results of this preliminary study suggest that PP Fusion primer set is better optimized for the analysis of ltDNA samples, and it is more robust regarding presence of the potential PCR inhibitors.

One of the genes considered as a risk factor for coronary artery disease (CAD) is the angiotensin-converting enzyme (ACE) gene. Many studies have been published regarding the relation between the ACE gene insertion/deletion (I/D) polymorphism and CAD. However, studies have provided controversial results. To explore this further in the population of Bosnia and Herzegovina, we compared the ACE I/D genotypes and alleles distribution between two groups: 100 CAD patients and 100 healthy control subjects. The higher distribution of DD genotype (47.0%) and D allele (65.5%) were found in CAD patients compared to controls (DD 34.0%; D allele 51.0%). Genotype odds ratio, (DD + ID) on the II, was 2.471 (1.252 – 4.876; 95% CI; p < 0.05). This leads to the conclusion that the DD genotype of the ACE I/D polymorphism affects the risk for development of coronary artery disease in Bosnian population.

In order to assess the genetic purity of common buckwheat variety ‘Darja’ which is the most commonly produced variety of this crop in Bosnia and Herzegovina, 10 SSR markers have been used. Five samples have been collected from different production regions in B&H (Breza, Nisici Plateau, Ustikolina, Bihac and Bosanska Krupa) and compared to the reference ‘Darja’ sample obtained from an ex situ seed collection from Slovenia. Seven out of ten primer pairs used managed to amplify SSR alleles. Analyses of molecular variance (AMOVA) showed a significant differentiation between the reference and all analyzed ‘Darja’ samples. Furthermore, the factorial correspondence analysis revealed a clear differentiation between the reference and ‘Darja’ samples from the most known production regions of common buckwheat in B&H clustering four out of five analyzed samples very close together. The most divergent one among the analyzed samples was the one from Ustikolina. Genetic purity of varieties of all of cross pollinated species produced in Bosnia and Herzegovina is questionable due to the general use of farm-saved seeds.

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.

The Dinaric endemic plant species Moltkia petraea (Tratt.) Griseb. is often called a "living fossil" of ancient Tertiary flora, with great importance for Bosnia and Herzegovina’s biodiversity. Considering its narrow and limited distribution range, insufficient data on the molecular background of this species is given so far. Due to the presence of various secondary metabolites that interfere with the DNA, isolation of nucleic acids from plant cells is known to be challenging. Even in closely related species it is necessary to optimize DNA isolation protocol in order to obtain high quality PCR amplifiable DNA. We collected 91 samples from five populations in Herzegovina. Doyle and Doyle (1987) CTAB protocol was modified by adding vitamin C (ascorbic acid) to the cell lysis buffer to improve DNA yield and quality. trnL(UAA) intron and nrDNA (ITS1, ITS2) molecular markers were applied to demonstrate amplifiability of isolated DNA and elucidate the intra- and interpopulation genetic diversity. Our results suggest a successful PCR amplification for 81% of the analyzed samples. PCR-RFLP analysis of trnL(UAA) revealed that all individuals in five populations have the same haplotype based on the obtained enzymatic profile for three enzymes (TaqI, HinfI, HindII). Alignment and comparison of ITS sequences didn’t reveal any hypervariable portion that could be informative in elucidating the genetic diversity of M. petraea populations. Further studies with additional application of microsatellite loci, RAPD and AFLP methods are necessary in an attempt to get insights into the genetic diversity of M. petraea.

This paper presents development of Artificial Neural Network (ANN) for prediction of the size of nanoparticles (NP) and microspore surface area (MSA). Developed neural network architecture has the following three inputs: the concentration of the biodegradable polymer in the organic phase, surfactant concentration in the aqueous phase and the homogenizing pressure. Two-layer feedforward network with a sigmoid transfer function in the hidden layer and a linear transfer function in the output layer is trained, using Levenberg-Marquardt training algorithm. For training of this network, as well as for subsequent validation, 36 samples were used. From 36 samples which were used for subsequent validation in this ANN, 80,5% of them had highest accuracy while 19,5% of output data had insignificant differences comparing to experimental values.

Cervical cancer represents a serious health problem affecting women worldwide especially in developing countries due to low socioeconomic status, inadequate health-care infrastructure, weaknesses in education on this particular issue and lack of effective screening programmes. The primary aim of this study was to assess alternative screening method for the improvement of cervical cancer prevention in conditions of Bosnia and Herzegovina (B&H), which could be applicable in other developing countries as well. The study was conducted on 101 subjects who provided their self-sampled vaginal swabs and/or cervical specimens collected by their gynecologists. Universal Human Papilloma Virus (HPV) primer set optimized to detect a wide range of HPV types was used for HPV genotyping from obtained swab samples in multiplex PCR. Amplicons were analyzed in agarose gel and Agilent 2100 bioanalyzer – a platform based on microfluid technology. Inter-rater agreement kappa (MedCalc2) was used to assess concordance between results of cervical and vaginal sample analysis. Out of 39 subjects who provided their vaginal and cervical samples, results of HPV detection mismatched in 10% of the cases. Inter-rater agreement showed good strength of coincidence between the results of cervical and vaginal sample analysis (kappa=0,748, CI=95 %). We presented an alternative PCR method for the detection of HPV based on vaginal self sampling which is affordable, informative, simple and applicable with high coverage level of defined targeted population and potentially significant in the given cultural and socioeconomic context.

Soy sauce is worldwide popular condiment of Asian origin. With the advent of GM soybean production, soy sauce drew the interest of food safety control. Samples collected for inspection are generally of industrial grade soy sauce type, which is produced from hydrolyzed soybean and grain. Following the failure to perform RealTime PCR based GMO screening on a number of submitted samples we tested our screening system on soy sauce produced following traditional method based on fermentation. Four batches of soy sauce were produced and DNA extracted. DNA concentration ranged from 32,68 to 65,36 ng/μl. Amplification of taxon specific target was successful with rather high Ct ( > 30). Promoter P-35S sequence was not detected, but T-NOS was detected in three samples with values reaching or exceeding LOD of the method. The results show that it is possible to detect transgenic elements in traditionally produced soy sauce while DNA extraction from industrial grade soy sauce is not possible.

The focus of this study was microsatellite diversity of crossbred horses raised in Bosnia and Herzegovina. Genomic DNA was extracted from blood samples of 20 individuals (KBA group – 7 individuals, crosses between Bosnian and Herzegovinian mountain horse and Arabian horse; KBR group – 9 individuals, crosses between Bosnian and Herzegovinian mountain and Belgian horses, crosses between Bosnian and Herzegovinian mountain horses and Holstein, crosses between Bosnian and Herzegovinian mountain and Lipizzaner horses and KBN group – 4 individuals, crosses between Bosnian and Herzegovinian mountain horse with an unknown origin of the other parent). The samples were profiled using 17 microsatellite markers. This method consisted of multiplex PCR procedure and generated reasonable amplification across all the loci. All samples were genotyped successfully. Considering all the observed parameters, VHL20 locus showed the highest microsatellite diversity. Locus HMS7 was the least variable in KBR group, while HMS1 locus was the least diverse in KBN group. The highest microsatellite diversity in KBA group was found at AHT5 locus while HTG6 locus was the least diverse. Obtained results suggest that the investigated populations of crossbred horses from Bosnia and Herzegovina are not affected by substantial loss of genetic diversity, as indicated by the presence of reasonably high level of genetic variation. An increase in the inbreeding coefficient and sufficient heterogenity in KBN group indicate occurrence of consanguineous mating. The present research contributes to the knowledge of current status of genetic structure of the investigated crossbred horses.

Atmospheric pollution is among the largest anthropogenic impacts on the ecosystem. In numerous studies it was observed that plants, especially those that grow in urban areas, are heavily influenced by different pollutants and their survival is correlated with structural and metabolic adaptation to stressful environmental conditions. Primary objective of this study was to determine the index of tolerance to air pollution (APTI) of plantain (Plantago major), on two locations in Zenica. The results indicated that index of tolerance to air pollution of P. major, APTI, is higher in individuals sampled from the contaminated site, than those in the control area.