This study compares the results obtained using two multiplex systems, PowerPlex® 16 System and PowerPlex® Fusion System, to evaluate the probability of a specific kinship relationship between the offspring of three pairs of identical twins, such as full kinship (siblings), first-degree relatives (first cousins) and half-siblings. Genomic DNA was isolated and amplified from buccal swab and selected short tandem repeat (STR) markers were detected. Electropherograms were generated and analyzed for all persons, using two multiplex systems. Paternity testing for every nine offspring of six examined couples was performed and in all cases the probability that the alleged father is the true father, was over 99.9999%. Kinship analyses were performed setting up two different hypotheses and calculating the likelihood ratio (LR) and kinship probability. Determining the degree of kinship between persons who were full siblings, likelihood ratio showed the highest values contrary to other two types of kinship. Kinship analyses between first cousins showed a higher probability that the examined persons are half-siblings, rather than they are first cousins. In most cases, the introduction of additional seven loci included in PowerPlex® Fusion System increased the values of average likelihood ratios. It is recommendable to use over 20 STR loci in complex kinship analyses.

The floods in Bosnia and Herzegovina in May 2014 caused landslides all over the country. In the small village of Šerići, near the town of Zenica, a landslide destroyed the local cemetery, relocated graves, and commingled skeletal remains. As the use of other physical methods of identification (facial recognition, fingerprint analysis, dental analysis, etc.) was not possible, DNA analysis was applied. DNA was isolated from 20 skeletal remains (bone and tooth samples) and six reference samples (blood from living relatives) and amplified using PowerPlex® Fusion and PowerPlex®Y23 kits. DNA profiles were generated for all reference samples and 17 skeletal remains. A statistical analysis (calculation of paternity, maternity, and sibling indexes and matching probabilities) resulted in 10 positive identifications. In this study, 5 individuals were identified based on one reference sample. This has once again demonstrated the significance of DNA analysis in resolving the most complicated cases, such as the identification of commingled human skeletal remains.

FOREWORD The science of the 21st century has not given up on its rapid development. The pace of evident progress in certain scientifc disciplines, especially those relying on applied genetics, does not allow for a breakthrough in the collection, sorting and presentation of the latest achievements made in hundreds of laboratories around the world. Continuous education of scientists, professors, experts, and users of scientifc achievements has never been this prominent and observable. After a brief analysis of the development of forensic genetics in the past fve years, we have decided that it would be wise to approach the complementation of existing material available with, as we then thought, “some new information”. But when we included everything we wanted to add onto the previous edition, we found that the new facts, hypotheses and models have been generated, as well as a promising direction for potential development established. Soon upon this realization, we had nothing left to do but to, signifcantly infuenced by young and enthusiastic associates, “roll up our sleeves” and prepare a new textbook. As a result, this book was created, which at the moment of its creation is probably the only existing edition that includes the most up-to-date information, especially related to the new multiplex STR systems, next-generation sequencing platforms and lineage markers, as well as new approaches in forensic DNA analysis in general. Two completely new chapters have been prepared, including the topics of food forensics and microbiology in forensic investigations. We are especially proud of the last chapter of this book that gives brief, understandable and highly applicable guidelines for proper sample handling, collection and storage, and overall model of behavior at the crime scene. As in the previous editions of this material, we tried to present the basic molecular biological, biochemical, statistical and technological knowledge, and other principles that must be known in order to comprehend the application of fundamental scientifc knowledge in forensic genetics. Also, we aimed at adding everything that is important into this book, and also what is written within the best books of the world, and everything that we have learnt from our practical work in the past decade. By preparing this edition in English language, we have thought of potential international readers of our book and tried our best to make this text as accessible worldwide as possible.

Tekst udžbenika podijeljen je u tri dijela: Opci dio, Specijalni dio te Izdvojena poglavlja. Sukladno ubrzanom tehnoloskom razvoju i sve vecim potrebama klinicke medicine i srodnih struka, neka poglavlja su prosirena najsvježijim podacima, a dodano je i vise potpuno novih poglavlja. Sva se poglavlja nalaze unutar podrucja određene medicinske patologije, a svako je popraceno recentnim literaturnim podacima. Sva poglavlja iznjedrila je klinicka praksa, temeljena na najnovijim spoznajama medicinske i biokemijske struke, kao i razvoj novih metoda operativne i „konzervativne“ medicine u posljednjih desetak godina. Dodatak udžbeniku je poglavlje Izvjestavanje o rezultatima medicinskobiokemijskih pretraga, koje sadrži Referentne intervale i preporucene vrijednosti u opcoj medicinskoj biokemiji s tri potpoglavlja: Laboratorijska hematologija, Laboratorijska koagulacija i Klinicka biokemija. U istom poglavlju su prikazane i Kriticne vrijednosti laboratorijskih nalaza. Drugo potpoglavlje odnosi se referentne intervale, preporucene vrijednosti, terapijska podrucja i toksicne koncentracije u specijalnim podrucjima medicinske biokemije i laboratorijske medicine. U knjizi je koristen klinicki pristup razlicitim patoloskim stanjima temeljen na analitickim rezultatima raznovrsnih dijagnostickih metoda sofisticirane medicinskolaboratorijske dijagnostike. Izdvojena poglavlja su: Laboratorij uz krevet bolesnika (POCT), Farmakogenetika, Toksikologija, Droge, Doping, Forenzicna analiza DNA. Naglasak knjige je na važnosti multidisciplinarnog rada, ali i potrebi primjene novih metoda moderne tehnologije u službi laboratorijske medicine te osiguranja kontrole kvalitete raznovrsnih dijagnostickih postupaka.

Abstract Background Posttraumatic stress disorder is characterized by an overactive noradrenergic system conferring core posttraumatic stress disorder symptoms such as hyperarousal and reexperiencing. Monoamine oxidase A is one of the key enzymes mediating the turnover of noradrenaline. Here, DNA methylation of the monoamine oxidase A gene exonI/intronI region was investigated for the first time regarding its role in posttraumatic stress disorder risk and severity. Methods Monoamine oxidase A methylation was analyzed via direct sequencing of sodium bisulfite-treated DNA extracted from blood cells in a total sample of N=652 (441 male) patients with current posttraumatic stress disorder, patients with remitted posttraumatic stress disorder, and healthy probands (comparison group) recruited at 5 centers in Bosnia-Herzegovina, Croatia, and the Republic of Kosovo. Posttraumatic stress disorder severity was measured by means of the Clinician-Administered Posttraumatic Stress Disorder Scale and its respective subscores representing distinct symptom clusters. Results In the male, but not the female sample, patients with current posttraumatic stress disorder displayed hypermethylation of 3 CpGs (CpG3=43656362; CpG12=43656514; CpG13=43656553, GRCh38.p2 Assembly) as compared with remitted Posttraumatic Stress Disorder patients and healthy probands. Symptom severity (Clinician-Administered Posttraumatic Stress Disorder Scale scores) in male patients with current posttraumatic stress disorder significantly correlated with monoamine oxidase A methylation. This applied particularly to symptom clusters related to reexperiencing of trauma (cluster B) and hyperarousal (cluster D). Conclusions The present findings suggest monoamine oxidase A gene hypermethylation, potentially resulting in enhanced noradrenergic signalling, as a disease status and severity marker of current posttraumatic stress disorder in males. If replicated, monoamine oxidase A hypermethylation might serve as a surrogate marker of a hyperadrenergic subtype of posttraumatic stress disorder guiding personalized treatment decisions on the use of antiadrenergic agents.

The relationship between genetic risk factors of thrombophilia and pregnancy loss (PL) is being discussed. The focus has been on F5 1691G>A, F2 20210G>A, and MTHFR 677C>T polymorphisms that may predispose women to microthrombosis during the stages of embryo implantation and placentation. Although, the frequencies of these polymorphisms were reported in different populations, such studies have not yet been performed in Bosnian population. In this study, we determined the prevalence of F5 G>A (rs6025), F2 G>A (rs1799963) and MTHFR C>T (rs1801133) polymorphisms in Bosnian women. A total of 154 women with PL, mean age 33 (±5.4) years, were enrolled in the study. As a control group, 154 mothers [mean age 31.4 (±6.7) years] with at least one live-born child were included. We used real-time polymerase chain reaction (PCR) to determine the frequencies of F5 G>A and F2 G>A genotypes, and PCR-restriction fragment length polymorphism (RFLP) for analyzing MTHFR C>T genotypes. The frequency of heterozygotes for F5 and F2 was significantly higher in women with venous thrombosis (VT) compared to women without VT (p = 0.047 and p = 0.001, respectively). There was no significant difference in the distribution of MTHFR genotypes and alleles between these two groups. In addition, we observed no significant differences in the genotype and allele frequencies between the group with PL and control group, for all investigated polymorphisms. The allele frequencies for 1691A (F5), 20210A (F2), and 677T (MTHFR) reported in this study are consistent with the data obtained for other European countries, however, we were not able to confirm the association between the three polymorphisms and PL in Bosnian women.

Widely considered as one of the cradles of human civilization, Mesopotamia is largely situated in the Republic of Iraq, which is also the birthplace of the Sumerian, Akkadian, Assyrian and Babylonian civilizations. These lands were subsequently ruled by the Persians, Greeks, Romans, Arabs, Mongolians, Ottomans and finally British prior to the independence. As a direct consequence of this rich history, the contemporary Iraqi population comprises a true mosaic of different ethnicities, which includes Arabs, Kurds, Turkmens, Assyrians, and Yazidis among others. As such, the genetics of the contemporary Iraqi populations are of anthropological and forensic interest. In an effort to contribute to a better understanding of the genetic basis of this ethnic diversity, a total of 500 samples were collected from Northern Iraqi volunteers belonging to five major ethnic groups, namely: Arabs (n = 102), Kurds (n = 104), Turkmens (n = 102), Yazidis (n = 106) and Syriacs (n = 86). 17-loci Y-STR analyses were carried out using the AmpFlSTR Yfiler system, and subsequently in silico haplogroup assignments were made to gain insights from a molecular anthropology perspective. Systematic comparisons of the paternal lineages of these five Northern Iraqi ethnic groups, not only among themselves but also in the context of the larger genetic landscape of the Near East and beyond, were then made through the use of two different genetic distance metric measures and the associated data visualization methods. Taken together, results from the current study suggested the presence of intricate Y-chromosomal lineage patterns among the five ethic groups analyzed, wherein both interconnectivity and independent microvariation were observed in parallel, albeit in a differential manner. Notably, the novel Y-STR data on Turkmens, Syriacs and Yazidis from Northern Iraq constitute the first of its kind in the literature. Data presented herein is expected to contribute to further population and forensic investigations in Northern Iraq in particular and the Near East in general.

Introduction: Single nucleotide polymorphisms (SNPs) have lately been used for prediction of metabolic processes that may be related to obesity. The aim of our study was to examine the association of SNPs of several genes with obesity and physical activity in 18 healthy volunteers. Methods: We used buccal swabs to collect and extract DNA from 18 volunteers. Pyrosequencing was used for molecular analysis of 13 polymorphisms in 10 genes (APOA2, MTHFR, MCM6, peroxisome proliferators-activated receptor gamma, FABP2, beta-2-adrenergic receptor (ADRB)2, ADRB3, A-actinin-3, angiotensin-converting enzyme, and FUT2). The volunteers’ personal data included body mass index (BMI), dietary practice and information on daily fitness and workout routine. Association between the 13 observed gene polymorphisms and individual BMI status (normal or overweight) was analyzed. Results of the DNA analysis were used for the expert evaluation by nutritionists and physiologists to obtain optimal regulation of nutrition and exercise. The volunteers had a dietary and fitness program for 12 months which they tracked by filling in a suitable study form. Results: 14 volunteers had a moderate genetic predisposition for abdominal adipose-tissue accumulation, while 4 of them had genotypes not associated with abdominal fat tissue accumulation. A statistically significant difference was found between the value of BMI before and after the implementation of personalized training and nutrition plan within the group of overweight volunteers (paired sample t=3.382; p = 0.006; exact p = 0.015). The single-locus F-test showed no association between the gene polymorphisms and BMI values. In addition, no correlation was detected between the gene polymorphisms and amount of BMI reduction prior and after the implementation of the personalized training and nutrition plan within the overweighed group of volunteers. Conclusion: Optimal nutrition and training plan are crucial for the BMI reduction as observed in the overweighed volunteers after the 12-month personalized training and individualized nutrition plan. However, the analyzed polymorphisms were not significantly associated with the obesity in this study.

Abstract Background: Tuzla Canton is the most populated region in the ethnically mixed territory of Bosnia and Herzegovina, whose genetic analysis could provide an insight into past demographic events. Aim: Analysis of 23 Y-chromosome STR markers in the population of Tuzla Canton and investigation of the genetic relationship of the male population of the Tuzla Canton and that of the larger Bosnian and Herzegovinian population as well as neighbouring and other European populations. Subjects and methods: The study was conducted among 100 unrelated healthy adult males from Tuzla Canton that have been genotyped using 23 Y-STR loci included in the PowerPlex Y23 kit. Statistical parameters such as haplotype diversity and allele frequencies were calculated, as well as the Rst-based genetic distances between the new dataset and those from Bosnia and Herzegovina and elsewhere, which were then visualised through multi-dimensional scaling plot and neighbour-joining phylogenetic tree analyses. Results: The PowerPlex Y23 kit has shown high discrimination capacity, as all 100 individuals have unique haplotypes. The newly incorporated loci seem to be highly informative. Population comparison reveals no statistically significant differences between the study population and the general Bosnian-Herzegovinian population, and between the study population and neighbouring populations. Conclusion: These results could be used as an additional investigation of the genetic relationship between the regional populations in Bosnia and Herzegovina and neighbouring human populations, as well as for further human population and forensic genetics studies.