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Conventional screening and diagnostic procedures in prostate complaints rely on PSA (Prostate Specific Antigen) concentration which is not specific for prostate cancer and frequently leads to unnecessary invasive procedures in order to exclude malignant disease. It is estimated that approximately 50% of persons who underwent tissue biopsy did so based on false positive PSA value. Therefore a proper and timely differential diagnosis of malignant disease using non-invasive techniques remains one of the biggest challenges in medicine. Urine is the invaluable source of biological information contained in small molecules i.e. RNA that is easily accessible and detectable using molecular genetics techniques. We describe economical and fast method for relative expression analysis applicable to any target gene using urine as a sample. Efficient non-invasive method for identification of malignant or high risk cases prove useful in reduction of patient distress during the diagnostic procedure and significantly reduce healthcare costs.

Sabina Vinković, K. Bajrovic, L. Pojskić

Genetics, as a science of the future, is very important part of Biology teaching. Within educational system in Bosnia and Herzegovina it is studied in limited number of clasess with a minimum of practical excercises, in both elementary and secondary schools. Limited understanding of scientific facts result in prejudices or uninformed attitudes towards novel biotechnologies that are largely based on non-reviewed and non-scientific information often incorrectly or inadequately presented in mass-media. Purpose of this research was to estimate the level of knowledge about selected topics in genetics among high school students in three Mostar high school institutions and differences in the level of knowledge between students from different high school programs – grammar school and vocational medical school. The research was conducted in 2017 among students of fourth grade of three high school institutions: ''Grammar School Mostar'', ''Medical high school Mostar'' and Medical high school ''Sestre milosrdnice''. Notable differences in knowledge level between two vocational programs are observed as well as variance in learning outcomes of the same program presented in two formal languages in Bosnia and Herzegovina

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.

Thomas W. Mühleisen, C. Reinbold, A. Forstner, L. Abramova, M. Alda, Gulja Babadjanova, M. Bauer, P. Brennan et al.

Cris E. Hughes, Mary P Rogers, A. Owings, Barbara Petzelt, Joycelynn Mitchell, Harold Harry, Theresa Williams, D. L. Goldberg et al.

This study presents genetic data for nine Native American populations from northern North America. Analyses of genetic variation focus on the Pacific Northwest (PNW). Using mitochondrial, Y chromosomal, and autosomal DNA variants, we aimed to more closely address the relationships of geography and language with present genetic diversity among the regional PNW Native American populations. Patterns of genetic diversity exhibited by the three genetic systems were consistent with our hypotheses: genetic variation was more strongly explained by geographic proximity than by linguistic structure. Our findings were corroborated through a variety on analytic approaches, with the unrooted trees for the three genetic systems consistently separating inland from coastal PNW populations. Furthermore, analyses of molecular variance support the trends exhibited by the unrooted trees, with geographic partitioning of PNW populations (FCT = 19.43%, p = 0.010 ± 0.009) accounting for over twice as much of the observed genetic variation as linguistic partitioning of the same populations (FCT = 9.15%, p = 0.193 ± 0.013). These findings demonstrate a consensus with previous PNW population studies examining the relationships of genome-wide variation, mitochondrial haplogroup frequencies, and skeletal morphology with geography and language.

Guy E. Boeckxstaens, V. Drug, D. Dumitrascu, A. Farmer, Johann Hammer, Trygve Hausken, B. Niesler, Daniel Pohl et al.

Introduction: Progressive pseudorheumatoid dysplasia (PPD) is an autosomal recessive genetic disorder reported to be caused by gene alterations of the Wnt1-inducible signaling pathway protein 3 corresponding gene (WISP3) located on chromosome position 6q22.  Up to date, there is only a handful of WISP3 mutations identified in Europe, whereas most mutations are identified in Asia and Middle East.  According to our knowledge, this is the first report of genetic dissection of WISP3 associated with spondyloepiphyseal dysplasia tarda from Bosnia and Herzegovina. Based on clinical examination findings (general manifestations, physical examination, characteristics of their bones on X-ray and laboratory results), an index patient was directed to WISP3 genotyping for confirmation of suspected diagnosis of PPD.Methods: DNA was extracted from peripheral blood leukocytes. All 5 exons and their exon-intron boundaries of the WISP3 gene were amplified by polymerase chain reaction (PCR) and sequenced by Sanger method. Segregation analysis was done to confirm the familial carrier status.Results: A missense mutation (C223G) - homozygous T to G transition at c.667 in exon 4 was identified in index patient. This mutation changed codon CAG to TAG and resulted in a subsequent change of the cysteine to glycine codon. Same mutation was observed in both parents in heterozygous form confirming the familial segregation.Conclusion: Due to its nature, the identified mutation C223G in exon 4 in WISP3 gene is the most probably causative for PPD in described patient. Here we describe the PCR based method for genotyping of specific mutation in WISP3 gene. The identification of this mutation might be a valuable addition to a regional databases on rare genetic variant although a functional analysis should be performed to explain its pathological effect.

Objectives: This study investigated the correlation between Tannerella forsythia, Porphyromonas gingivalis, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans at dual sites in concurrent endodontic-periodontal diseases. Material and methods: Samples were collected from endodontium and periodontium in cases of concurrent endodontic-periodontal diseases from thirty participants. The sensitivity and specificity of SYBR Green real-time PCR was used to identify the targeted species. Absolute number of targeted genome copies in tested samples were extrapolated from respective calibration curve. Results: No statistical difference was found in the number of detected endodonticperiodontal pathogens between the endodontium and periodontium. The Pearson test detected significant correlation (P<0.001) between targeted bacteria; T. forsythia, F. nucleatum, and P. gingivalis from endodontic-periodontal lesions. Synergistic component observed separately in endodontic biofilm was found only between T. forsythia and F. nucleatum (r=0.380, P=0.03) while in periodontal biofilm T. forsythia, F. nucleatum and P. gingivalis gave high synergism result (P<0.0001). Correlation analysis showed that T. forsythia in primary endodontic infection and in periodontal lesion was significantly decreased with the increase of patients age (r=-0.308, P=0.017). Conclusions: Correlation between targeted bacterial species levels from concurrent endodontic-periodontal diseases confirmed that coronal and cervical dentinal tubules may represent a viable pathway that allows spreading and maintaining of dual sites infection. Periodontal bacteria detected in root canal of concurrent endodonticperiodontal infections may originate from the local periodontal lesions.

Introduction: Progressive pseudorheumatoid dysplasia (PPD) is an autosomal recessive genetic disorder reported to be caused by gene alterations of the Wnt1-inducible signaling pathway protein 3 corresponding gene (WISP3) located on chromosome position 6q22. Up to date, there is only a handful of WISP3 mutations identified in Europe, whereas most mutations are identified in Asia and Middle East. According to our knowledge, this is the first report of genetic dissection of WISP3 associated with spondyloepiphyseal dysplasia tarda from Bosnia and Herzegovina. Based on clinical examination findings (general manifestations, physical examination, characteristics of their bones on X-ray and laboratory results), an index patient was directed to WISP3 genotyping for confirmation of suspected diagnosis of PPD. Methods: DNA was extracted from peripheral blood leukocytes. All 5 exons and their exon-intron boundaries of the WISP3 gene were amplified by polymerase chain reaction (PCR) and sequenced by Sanger method. Segregation analysis was done to confirm the familial carrier status. Results: A missense mutation (C223G) homozygous T to G transition at c.667 in exon 4 was identified in index patient. This mutation changed codon CAG to TAG and resulted in a subsequent change of the cysteine to glycine codon. Same mutation was observed in both parents in heterozygous form confirming the familial segregation. Conclusion: Due to its nature, the identified mutation C223G in exon 4 in WISP3 gene is the most probably causative for PPD in described patient. Here we describe the PCR based method for genotyping of specific mutation in WISP3 gene. The identification of this mutation might be a valuable addition to a regional databases on rare genetic variant although a functional analysis should be performed to explain its pathological effect.

There are two major theories for inheritance of Rh blood group system: Fisher - Race theory and Wiener theory. Aim of this study was identifying frequency of RHDCE alleles in Bosnian - Herzegovinian population and introduction of this method in screening for Rh phenotype in B&H since this type of analysis was not used for blood typing in B&H before. Rh blood group was typed by Polymerase Chain Reaction, using the protocols and primers previously established by other authors, then carrying out electrophoresis in 2-3% agarose gel. Percentage of Rh positive individuals in our sample is 84.48%, while the percentage of Rh negative individuals is 15.52%. Inter-rater agreement statistic showed perfect agreement (K=1) between the results of Rh blood system detection based on serological and molecular-genetics methods. In conclusion, molecular - genetic methods are suitable for prenatal genotyping and specific cases while standard serological method is suitable for high-throughput of samples.

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