Diversity of nuclear microsatellite markers were analyzed in a reference sample of the population of northeast Bosnia and Herzegovina. 437 samples taken from unrelated individuals were processed and three samples of paternity proof were shown. Detection effectiveness profile of the research, points to a valid choice of method of extraction, amplification and genotyping short tandem repeat (STR) loci with PowerPlextm16 kit. Genetic analysis of allelic variants of the 15 STR loci PowerPlextm16 kit detected 17 samples determined as rare allelic variants or microvariants. Samples were divided into 15 different allelic variants at 7 different loci, and are: in locus D7S820, D16S539, D3S1358, D18S51, PENTA D, PENTA E and in locus vWA. Genetic analysis of mutations in cases of paternity determined three examples of single-step mutations in the loci FGA, Penta D and D3S1358. Genetic analysis of observed STR loci detected three allelic variant of genotype combination 7/10/11.3 in locus D7S820 Type II. Population genetic analysis of STR loci in a representative sample of the population of northeast Bosnia and Herzegovina included the application of the assessment tests of within-population genetic diversity and interpopulation diversity, as well as genetic differentiation between populations: North-eastern Bosnia and Herzegovina (BH) and BH general reference, then the Croatian population, Macedonian, Serbian and Slovenian. Based on the result analysis of specific forensic parameters, it can be assumed that the most informative marker is PENTA E for population genetic analysis and forensic testing in the population of northeast Bosnia and Herzegovina. Research results fit regional STR database of this part of Europe.
The primary value of DNA profiling significantly increased over the last fifteen years due to introduction of short tandem repeat (STR) loci in routine paternity testing, forensic work and identification of victims of mass disaster. Different methods of forensic DNA testing (known as DNA fingerprinting) have been widely established and accepted as standard procedures of investigation. This lecture presents the experience of the last eighteen years of identification of missing persons in Bosnia and Croatia. The data show that current protocols and procedures optimized for relatively fresh bones and teeth can be used in analysis of much older samples without significant modification. Also, introduction of new technologies (e.g., miniSTR assay) can help recover information from degraded DNA samples that typically result in partial profiles and total loss of information from regular STR amplicons. This approach has already been used in the analysis of highly degraded samples like those from the victims of World Trade Center terrorist attacks and WWII. Now the use of this scientific approach has been proven even for the two millenia old bone samples. In collaboration with Roche Molecular Systems recently we used a new generation of mtDNA strips and were able to get full profiles from several century old bones even in a cases where STR amplification completely failed.
The European Network of Forensic Science Institutes (ENFSI) recommended the establishment of forensic DNA databases and specific implementation and management legislations for all EU/ENFSI members. Therefore, forensic institutions from Bosnia and Herzegovina, Serbia, Montenegro, and Macedonia launched a wide set of activities to support these recommendations. To assess the current state, a regional expert team completed detailed screening and investigation of the existing forensic DNA data repositories and associated legislation in these countries. The scope also included relevant concurrent projects and a wide spectrum of different activities in relation to forensics DNA use. The state of forensic DNA analysis was also determined in the neighboring Slovenia and Croatia, which already have functional national DNA databases. There is a need for a ‘regional supplement’ to the current documentation and standards pertaining to forensic application of DNA databases, which should include regional-specific preliminary aims and recommendations.
The aim of this article is to offer a concise interpretation of the scientific data about the topic of Croatian genetic heritage that was obtained over the past 10 years. We made a short overview of previously published articles by our and other groups, based mostly on Y-chromosome results. The data demonstrate that Croatian human population, as almost any other European population, represents remarkable genetic mixture. More than 3/4 of the contemporary Croatian men are most probably the offspring of Old Europeans who came here before and after the Last Glacial Maximum. The rest of the population is the offspring of the people who were arriving in this part of Europe through the southeastern route in the last 10 000 years, mostly during the neolithization process. We believe that the latest discoveries made with the techniques for whole-genome typing using the array technology, will help us understand the structure of Croatian population in more detail, as well as the aspects of its demographic history.
Background: The population of the island of Cres presents one of the few persisting Eastern Adriatic isolates and is thereby suitable for human population differentiation analyses. Aim: The aim of this study was to analyse the genetic structure of the island of Cres with respect to its eight sub-populations and to compare the genetic variation of the island of Cres with other Eastern Adriatic islands and the Croatian mainland. Subjects and methods: Fifteen AmpFlSTR identifiler loci were analysed in a sample group of 122 unrelated autochthonous individuals from the island of Cres, Croatia. Results: Analysis of STR polymorphisms revealed genetic homogeneity among sub-populations of the island of Cres and small but significant levels of genetic heterogeneity among geographically distant Eastern Adriatic islands. Conclusion: Despite a considerable degree of genetic homogeneity among the studied Eastern Adriatic islands, small but significant differentiation between distant islands indicates geographic sub-structuring which follows the isolation by distance model. This study is supportive of the notion that STR markers are useful for genetic differentiation between larger and geographically more distant regions.
Lilium martagon L. var. cattaniae Vis. (Liliaceae) is endemic plant of Dinaridi mountain. In this work we established protocol for fast in vitro propagation and multiplication of Lilium martagon var. cattaniae. The aim was to enable fast production of plant material as potential source of pharmaceutically valuable secondary metabolites. Seeds of L.martagon var. cattaniae were germinated on a Murashige and Skoog basal medium with a supplement of 0.15 mg/l gibberellic acid (GA3), and multiplication was performed on MS medium supplemented with 0.1 mg/l gibberellic acid (GA3), 0.2 mg/l indole-3-butyric acid (IBA) and 0.5 mg/l 6-ben- zylaminopurine (BAP). We used ultrasound assisted extraction to prepare extracts of leaves and bulbs of Lilium martagon var. cattaniae, which were evaluated for their genotoxic potential using Allium test and cytokinesis-block micronucleus test in human lymphocytes culture. There was statistically significant difference between all used concentrations of lilium extracts and control on proliferation of cells of root tip of onion (Allium cepa). In cytokinesis-block micronucleus test no statistically significant difference between frequencies of analyzed parameters in samples treated with tested concentrations of extracts and control was obtained.
Forensic parameters based on 15 AmpFISTR Identifiler short tandem repeat (STR) loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818 and FGA) were evaluated in the sample of 122 unrelated, autochthonous, adult individuals from the Island of Cres (Croatia). PCR amplification was performed with the AmpFISTR Identifiler PCR Amplification Kit and the amplified products were separated and detected using the ABI 3130 DNA genetic analyzer. The agreement with Hardy Weinberg Equilibrium (HWE) was confirmed for all loci (p > 0.05). The combined power of discrimination (PD) and the combined power of exclusion (PE) for the 15 tested STR loci were 0.99999999999999997988728679 and 0.999997397, respectively. According to the presented data, D18S51 proved to be the most informative marker followed by markers D2S1338 and D21S11. Interpopulation comparisons in allele frequencies with other East Adriatic Islands revealed significant differences for all analyzed population pairs ranging from 4 loci (Cres vs. Hvar) to 1 locus (Cres vs. Krk). Furthermore, allele frequencies comparisons of Cres and Croatian mainland revealed the lack of statistically significant differences at all studied loci. The results of the current study indicate that the examined fifteen STR loci are useful genetic markers for individual identification and paternity testing in Croatian population from the Island of Cres.
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