In this case report, we present the results of a complex kinship investigation involving six individuals, whose biological samples were submitted over an extended period with unclear and shifting instructions. Initially, the goal appeared to be a general reconstruction of familial relationships, yet the underlying motive remained unclear. As the genetic analyses progressed, new individuals were added, and the focus gradually shifted toward a concealed paternity issue. To clarify the familial connections among the tested individuals, we applied a combination of autosomal, X and Y-chromosomal Short Tandem Repeat (STR) markers, coupled with robust statistical methods. DNA was extracted from various personal items, and STR profiles were generated using validated forensic kits and software. Initial autosomal STR analysis primarily revealed familial relationships among three female and one male individual. The subsequent inclusion of two additional individuals not only supported the initial findings but also revealed a previously concealed paternity relationship between two of the subjects. Further analysis using X and Y-STR markers verified the autosomal STR analysis results, thereby supporting the previously established familial relationships. Despite issues like poor DNA quality in certain samples and insufficient contextual data provided, the genetic data allowed for high-confidence resolution of multiple kinship ties, including the confirmation of full- and half-sibling relationships, maternal links, and paternity. This case underscores the efficacy of molecular-genetic tools in revealing concealed familial structures and resolving disputed paternity within complex forensic contexts.

With the aim to improve the present X-STR database regarding the population of Bosnia and Herzegovina (B&H), we report the genetic data from 228 unrelated adults residing in different regions of B&H that were genotyped using the QIAGEN's Investigator® Argus X-12 kit which detects 12 STR markers distributed over the entire X-chromosome as four distinct linkage groups. Our results indicate that the 12 X-STR loci examined are highly polymorphic in the B&H population. Distribution of allele frequencies, calculated only for female samples, did not show significant deviation from Hardy-Weinberg equilibrium (p > 0.05). Moreover, PIC values for all analyzed X-STR loci and sample groups proved to be high (PIC≥0.6); the most informative X-STR marker is DXS10135 (for male, female and pooled samples: 0.918647, 0.912071 and 0.918621, respectively) and the most informative linkage group is LG I (PIC = 0.988903076). Also, B&H population was compared to other 14 populations; no significant differences were observed between B&H and neighboring populations of Serbia, Slovenia and Croatia. Contrariwise, the greatest genetic distance was observed in comparison with South African population (0.1449). Overall, this paper is the first report of forensically relevant parameters, allele and haplotype frequencies for the 12 X-STR loci included in the Investigator® Argus X-12 marker system in the population of Bosnia and Herzegovina. Also, this study strongly supports the application of X-STR markers for human identification, provides new insights into the genetic structure of contemporary population and contributes to the existing database of B&H population.

Numerous archaeological sites in Bosnia and Herzegovina represent a historical heritage and testify to the rich cultural, social, and political life of medieval Bosnia. Bobovac, the capital of the Bosnian Kingdom after King Tvrtko I's coronation in 1377, featured a royal complex with a palace, church, and fortification. Recent molecular-genetic research on skeletal remains from Bobovac aims to uncover medieval ancestors' customs and genetic origins. Fifteen well-preserved teeth samples from Bobovac were processed. STR amplification employed PowerPlex® Fusion and Investigator® 24plex QS Kits, with Y-STR profiles generated using the PowerPlex® Y23 System. Fourteen partial autosomal STR profiles were obtained, enabling sex determination and kinship analysis. STR amplification success varied due to ancient DNA degradation, with larger loci showing lower amplification rates. Kinship analysis confirmed appropriate marker selection, demonstrating high reliability for determining close relationships. Integrating aDNA analysis with archaeological research enhances our understanding of historical populations, connecting archaeology and forensic genetics to contribute to the broader narrative of human history.

Many heavy metals (HMs) are essential micronutrients for the growth and development of plants. However, human activities such as mining, smelting, waste disposal, and industrial processes have led to toxic levels of HMs in soil. Fortunately, many plant species have developed incredible adaptive mechanisms to survive and thrive in such harsh environments. As a widespread and ruderal species, Geranium robertianum L. inhabits versatile soil types, both polluted and unpolluted. Considering the ubiquity of G . robertianum , the study aimed to determine whether geographically distant populations can tolerate HMs. We collected soil and plant samples from serpentine, an anthropogenic heavy metal contaminated, and a non-metalliferous site to study the physiological state of G. robertianum . HMs in soil and plants were determined using flame atomic absorption spectrometry. Spectrophotometric methods were used to measure the total content of chlorophylls a and b, total phenolics, phenolic acids, flavonoids, and proline. Principal component analysis (PCA) was used to investigate the potential correlation between HMs concentrations gathered from various soil types and plant samples and biochemical data acquired for plant material. A statistically significant difference was observed for all localities regarding secondary metabolite parameters. A positive correlation between Ni and Zn in soil and Ni and Zn in plant matter was observed ( p <0.0005) indicating higher absorption. Regardless of high concentrations of heavy metals in investigated soils, G. robertianum displayed resilience and was capable of thriving. These results may be ascribed to several protective mechanisms that allow G. robertianum to express normal growth and development and act as a pioneer species.

Medieval cemeteries Klisa-Guca Gora, Alihodze and Glavica-Han Bila located in the Travnik area (Travnik, Bosnia and Herzegovina) were archaeologically examined in the period 2011-2014, revealing human skeletal remains of 11 individuals in total. Archaeological skeletal samples, previously deposited in Travnik Homeland Museum (Travnik, Bosnia and Herzegovina) were subjected to genetic analysis. The aim of this research was to test familiar relationship of 11 individuals excavated from three medieval cemeteries and to predict Y-haplogroup for male individuals. In order to perform molecular-genetic characterisation of collected human skeletal remains, two systems of genetic markers were analysed: autosomal and Y-STR loci. Complete or partial data obtained by autosomal STR typing of 11 individuals were subjected to kinship analysis. Male sex was determined in eight samples out of 11. Direct relatives of the "brother-brother" type were detected in one case with high kinship probability (KP) value of 99.99996 %. Complete or nearly complete and usable Y-STR profiles were obtained for six out of eight male individuals. The presence of identical haplotypes at Y-STR loci and results of Y-haplogroup prediction suggest that all male individuals share the same paternal lineage and belong to J2a haplogroup. Overall, this study emphasises the usefulness, efficiency and sensitivity of STR markers in the molecular-genetic characterisation of old skeletal remains as well as the importance of employing additional markers like Y-STRs in archaeogenetic studies, besides traditionally used autosomal STR markers, in order to get a comprehensive information about close and distant relatives, and ancestry.

Sexual violence represents a widespread social problem associated with serious lifelong consequences. In many cases, an outcome of sexual violence is the victim's unwanted pregnancy, usually ended in an abortion. The objective of this paper is to report five rape cases, including rapes of a minor and young woman, two incest cases and a case of human trafficking for sexual exploitation, where every case resulted in the victim's pregnancy. In each case, pregnancy was terminated in the first trimester or at the beginning of the second trimester in the relevant medical center or clinic. Fresh fetal blood or aborted tissue samples were delivered to our laboratory in order to perform paternity testing for the purpose of proving the crime. DNA extraction using Qiagen Dneasy™ Tissue Kit was optimized according to the sample type. Amplification of autosomal STR (Short Tandem Repeat) markers was performed using the PowerPlex®16 System. In two cases, mixtures of maternal and fetal DNA in the aborted fetal material were found. Using the LRmix Studio v.2.1.5 Software for interpreting DNA mixtures based on a probabilistic model, the likelihoods of maternal contribution and presence of fetal allelic variants inherited from the alleged father/suspect were calculated. Based on these results, we confirmed the presence of assumed fetal fractions (determined before software analysis) in the mixtures. In all cases, positive paternity proved the crime (probabilities of paternity >99.9999%). This cases report once again pointed out the importance of DNA analysis in the process of clarifying and solving forensic cases and demonstrated that the LRmix Studio v.2.1.5 Software can deal with complex cases such as sexual assaults.

During 2019, 28 grave constructions were excavated at cemetery Metaljica (Hadzici municipality, Bosnia and Herzegovina). Archaeological excavation have revealed 17 well preserved skeletons that were subjected to anthropological, odontological and genetic analyses. To determine sex by genetic analysis, amelogenin and DYS391 loci were taken into consideration. Concordance between results of anthropological, odontological and genetic analyses applied in this research occurred in one sample out of 17. In four samples, sex was determined only by genetic analysis, since sexual dimorphism indicators were not preserved. Concordance between odontological and genetic determinations was observed in two samples, while affirmative results of anthropological and genetic analysis were obtained in only one sample. Discrepancy in the majority of samples can be atributted to the state of preservation of skeletal remains, interpopulation differences and allele drop-out. Moreover, anthropological and odontological analyses were not applicable to juvenile/subadult skeletons, since sexually dimorphic characteristics relevant for metric and morphological analysis are not developed well at that developmental period. This study emphasizes the importance of combining anthropological, odontological and genetic methods in order to determine sex of archaeological skeletal remains accurately.

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.

During 2019, 28 grave constructions were excavated at cemetery Metaljica (Hadzici municipality, Bosnia and Herzegovina)

Abstract Clinopodium alpinum subsp. orontium (K.Malý) Govaerts and Thymus bracteosus Vis. ex Benth. are endemic Lamiaceae species in Bosnia and Herzegovina with rather limited data about their cytotoxic and genotoxic effects. This study aimed to analyse phenolic compounds composition of C. alpinum subsp. orontium and T. bracteosus aqueous and dimethyl sulphoxide (DMSO) extracts and their cytotoxic and genotoxic potential in human peripheral blood lymphocyte cultures. Among 33 analytes, 17 were identified and quantified in the examined extracts with the rosmarinic and chlorogenic acids as main constituents. Genotoxic effects of extracts from both species are proven at the highest applied dose. T. bracteosus extracts and DMSO as a solvent exhibited stronger genotoxic potential that should be further investigated in tumour cell lines. Nevertheless, non-endemic species with similar phenolic composition and bioactivity should be the first choice for medicinal purposes. Graphical Abstract

Abstract Background: Bosnia and Herzegovina is a multinational and multireligious country, located in the western part of the Balkan Peninsula. Migrations through history were a key factor in the genetic identity of the Bosnian–Herzegovinian population. Aim: To analyse genetic polymorphisms of 22 autosomal short tandem repeat (STR) loci in the population of Bosnia and Herzegovina and to compare STR allele frequencies for STR loci with the reference data for European populations. Subjects and methods: The study was conducted among 600 unrelated individuals from all regions of Bosnia and Herzegovina. Genotyping was performed using the PowerPlex® Fusion amplification kit. Allele frequencies and statistical parameters were calculated, as well as the genetic distance among analysed populations through the construction of a neighbor-joining dendrogram. Results: STR loci included in the PowerPlex® Fusion amplification kit showed high discriminatory power indicating their reliability for human identification and paternity testing. The neighbor-joining dendrogram based on the results of genetic distance analysis showed that the Bosnian and Herzegovinian population has the greatest genetic distance from Turkish and Hungarian populations and greatest similarity with Croatian, Slovenian, and Serbian populations. Conclusion: The results of this study strongly support the application of 22 autosomal genetic markers for paternity testing and personal identity testing and are in agreement with most previous human studies in the investigated human populations.