Availability of the large number of sources of extractable human DNA poses the challenge to find the easiest way to obtain large quantities of DNA from those sources. We have used Maxwell and Qiagen methods for DNA extractions from various samples that could be found at the crime scene or used in paternity disputes. Samples of buccal, vaginal and ear swabs, cigarette tips, chewing gum, urine, contact lenses, hair and dandruff were collected from ten voluntary donors. We used commercial Qiagen and Maxwell systems for DNA isolation. The success of isolation was demonstrated by horizontal gel electrophoresis, spectrophotometry analysis and PCR amplification of 15 STR loci (PowerPlex® 16 System). Our results confirmed that the Maxwell system was the fastest and easiest and a very reliable way of getting high-quality DNA from different sources. We have got 9 of 10 complete DNA profiles by this system, whereas DNA extraction according to Qiagen protocol resulted in generating only two out of ten completed DNA profiles. In conclusion, the Maxwell system appears to be the method of the choice for DNA extraction from large numbers of biological samples because of its reliability, simplicity and cost-effectiveness.

Aim To present the results obtained in the identification of human remains from World War II found in two mass graves in Ljubuški, Bosnia and Herzegovina. Methods Samples from 10 skeletal remains were collected. Teeth and femoral fragments were collected from 9 skeletons and only a femoral fragment from 1 skeleton. DNA was isolated from bone and teeth samples using an optimized phenol/chloroform DNA extraction procedure. All samples required a pre-extraction decalcification with EDTA and additional post-extraction DNA purification using filter columns. Additionally, DNA from 12 reference samples (buccal swabs from potential living relatives) was extracted using the Qiagen DNA extraction method. QuantifilerTM Human DNA Quantification Kit was used for DNA quantification. PowerPlex ESI kit was used to simultaneously amplify 15 autosomal short tandem repeat (STR) loci, and PowerPlex Y23 was used to amplify 23 Y chromosomal STR loci. Matching probabilities were estimated using a standard statistical approach. Results A total of 10 samples were processed, 9 teeth and 1 femoral fragment. Nine of 10 samples were profiled using autosomal STR loci, which resulted in useful DNA profiles for 9 skeletal remains. A comparison of established victims' profiles against a reference sample database yielded 6 positive identifications. Conclusion DNA analysis may efficiently contribute to the identification of remains even seven decades after the end of the World War II. The significant percentage of positively identified remains (60%), even when the number of the examined possible living relatives was relatively small (only 12), proved the importance of cooperation with the members of the local community, who helped to identify the closest missing persons’ relatives and collect referent samples from them.

The goal of this study was to examine the effectiveness of 6 STR markers application (D21S1435, D21S11, D21S1270, D21S1411, D21S226 and IFNAR) in molecular genetic diagnostics of Down syndrome (DS) and to compare it with cytogenetic method. Testing was performed on 73 children, with the previously cytogenetically confirmed Down syndrome. DNA isolated from the buccal swab was used. Previously mentioned loci located on chromosome 21 were simultaneously amplified using quantitative fluorescence PCR (QF PCR). Using this method, 60 previously cytogenetically diagnosed DS with standard type of trisomy 21 were confirmed. Furthermore, six of eight children with mosaic type of DS were detected. Two false negative results for mosaic type of DS were obtained. Finally, five children with the translocation type of Down syndrome were also confirmed with this molecular test. In conclusion, molecular genetic analysis of STR loci is fast, cheap and simple method that could be used in detection of DS. Regarding possible false results detected for certain number of mosaic types, cytogenetic analysis should be used as a confirmatory test.

Environmental conditions may have impact on plant metabolism, especially on secondary metabolism. As a result of different stress circumstance, plants have developed different protective mechanisms and major one is production of secondary metabolites. Plant growth conditions could be controlled and modified in in vitro plant culture, which usually results in higher or lower contents of secondary metabolites. We have established a rapid protocol for in vitro germination and cultivation of Brassica oleracea L. var. italica Plank. Three, ten, twenty and thirty days old seedlings, cultivated on three different Murashige-Skoog (MS) media, as well as two types of spontaneously induced calli were used for extraction. Ethanolic plant extracts were tested for their antioxidative potential using 2,2’-diphenyl-1picrylhydrazyl (DPPH) radical scavenging method. Extracts from three days old seedling demonstrated the highest antioxidative potential. On the other hand, extract of broccoli seedlings cultivated on basal MS medium have shown prooxidative properties that can be contribute to prooxidative properties of some unknown component in the presence of free transition metal ions, the type of oxidizable substrate in use, as well as to the biological environment in which they act. Article info Received: 06/12/2012 Accepted: 17/12/2012

Aim To detect polymorphisms of 23 Y-chromosomal short tandem repeat (STR) loci, including 6 new loci, in a reference database of male population of Bosnia and Herzegovina, as well as to assess the importance of increasing the number of Y-STR loci utilized in forensic DNA analysis. Methods The reference sample consisted of 100 healthy, unrelated men originating from Bosnia and Herzegovina. Sample collection using buccal swabs was performed in all geographical regions of Bosnia and Herzegovina in the period from 2010 to 2011. DNA samples were typed for 23 Y STR loci, including 6 new loci: DYS576, DYS481, DYS549, DYS533, DYS570, and DYS643, which are included in the new PowerPlex® Y 23 amplification kit. Results The absolute frequency of generated haplotypes was calculated and results showed that 98 samples had unique Y 23 haplotypes, and that only two samples shared the same haplotype. The most polymorphic locus was DYS418, with 14 detected alleles and the least polymorphic loci were DYS389I, DYS391, DYS437, and DYS393. Conclusion This study showed that by increasing the number of highly polymorphic Y STR markers, to include those tested in our analysis, leads to a reduction of repeating haplotypes, which is very important in the application of forensic DNA analysis.