Correlation of periodontal pathogens in concurrent endodontic-periodontal diseases

Objectives: This study investigated the correlation between Tannerella forsythia, Porphyromonas gingivalis, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans at dual sites in concurrent endodonticperiodontal 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 endodontic-periodontal pathogens between the endodontium and periodontium. The Pearson test detected significant correlation (P\u3c0.001) 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\u3c0.0001). 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 endodonticperiodontal 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 endodontic-periodontal infections may originate from the local periodontal lesions

Diversity of nuclear short tandem repeat loci in representative sample of North-eastern Bosnian and Herzegovina population

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

Overview of Human Population-Genetic Studies in Bosnia and Herzegovina during the Last Three Centuries: History and Prospective

Modern Bosnia and Herzegovina is a multi-ethnic and multi-religion country, with a very stormy history. Certain archaeological findings indicate continuous population of its territory since the Paleolithic. In time, vast number of different factors jointly influenced fascinating diversity of local human populations. A great number of small, more or less isolated, indigenous populations, make this area quite attractive for population-genetic surveys of different levels and approaches. Austro-Hungarian military physicians conducted the very first known bio-anthropological analyses of Bosnia-Herzegovina population at the end of the 19th century. Thus, the first step towards resolving the genetic structures of local B&H human populations was made. The studies that followed (conducted throughout most of the 20th century) were primarily based on the observation of various phenotypic traits. This stage was followed by the examination of various cytogenetic and fundamental DNA based molecular markers. The efforts undertaken over the last three centuries revealed »human genetic treasure« in Bosnia and Herzegovina. However, even now, after all the studies that were conducted, many interesting features remain to be discovered and described within the existing local human populations

Frequency of Main Western-Euroasian mtDNA Haplogroups and Paleolithic and Neolithic Lineages in the Genetic Structure of Population of Northeastern Bosnia

Mitochondrial DNA (mtDNA) variations were analyzed in a sample of 245 individuals of Bosnian-Herzegovinian population from the area of ​​ Northeastern Bosnia (also known as Tuzla region). Haplogroup affiliation was determined using RFLP method (Restriction Fragment Length Polymorphism) analyzing haplogroup-specific markers of mtDNA coding region, characteristic for the main Western-Eurasian haplogroups. Additional analyses of two sequenced hypervariable segments (HVSI and HVSII) of mtDNA control region were performed in order to identify U subhaplogroups. The study revealed that 95.51% of the analyzed individuals belonged to the typical Western-Eurasian haplogroups: H, I, J, K, T, U, V, W or X. The most frequent haplogroup in the analyzed population was the haplogroup H (52.65%) which, due to its increased frequency, represents a marking haplogroup of the population of Northeastern Bosnia. The results of intergroup genetic analysis showed that Bosnian-Herzegovinian population is genetically closer to previously studied populations of Herzegovinians (part of Bosnia and Herzegovina), Slovenians and Croats in relation to other neighboring populations located in Southeastern Europe. Our study also suggests that population genetic structure of Tuzla region is dominated by mutations that are classified as „Paleolithic”. These mutations were probably brought to the area of Northeastern Bosnia through waves of prehistoric and historic migrations, but the impact of any pre-Neolithic, Neolithic or some ,,later,, migrations, with a slightly lower contribution to the genetic structure of this population, also can-not be neglected

VALIDATION OF VAGINAL SELF-SAMPLING AS AN ALTERNATIVE OPTION IN PCR BASED DETECTION OF HPV IN CERVICAL CANCER SCREENING 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

MtDNA variations in genetic structure of populations in Tuzla region (Bosnia and Herzegovina)

MtDNA variations, determined in terms of mtDNA haplogroups, were analyzed, the sample consisted of the human population of Bosnia and Herzegovina from the teritory of the Tuzla region. The analysis included a total of 245 mtDNA samples (kin unrelated individuals). Determination of mtDNA haplogroups was studied by using haplogroup-specific PCR-RFLP markers of a coding region of mtDNA. We observed that 95.11% of investigated individuals in the Tuzla region belong to the Western-Euroasian mtDNA haplogroups: H, V, T, I, J, K, U, X, W. The impact of migratory activities on the population structure of the Tuzla region was estimated by analysis within- and of intergroup genetic differences (index of genetic specificities, pairwise FST analysis and exact-p test) of indigenous and migrant categories (recent, internal, and historical). The analysis showed that a significant demographic influence on the genetic structure of indigenous people was exerted by the category of recent migrants only

Assessment of relatedness between neurocan gene as bipolar disorder susceptibility locus and schizophrenia

Large scale genetic association meta-analyses showed that neurocan (NCAN) gene polymorphism rs1064395 is susceptibility locus for bipolar disorder. These studies also included patients with bipolar disorder originated from Bosnia and Herzegovina. Followed by theory of shared genetic elements between bipolar disorder and schizophrenia susceptibility, other studies explored several genetic factors with schizophrenia vulnerability as well. In this work, authors investigated the association between previously confirmed bipolar disorder genetic risk factor-neurocan with schizophrenia in a population sample of Bosnia and Herzegovina. Ethical aspects of this research were assessed by Ethics Committee of Clinical Center University of Sarajevo. Blood samples for DNA extraction were taken from the total of 86 patients and healthy individuals who previously signed informed consent. Genotyping for rs 1064395 was done using direct sequencing method. A case-control analysis of common genetic polymorphism within neurocan gene and schizophrenia status in a consecutively sampled patient cohort have been done using Fisher-exact test with odds-ratio calculation. No statistically significant allele and genotype association with disease status was found (p>0.05). Our finding supports the fact that large-scale genetic association studies approach need to be employed when detecting the variants with small additive effect in phenotypes with complex ethiology

Analysis of Delphinidin and Luteolin Genotoxicity in Human Lymphocyte Culture

Introduction: Bioflavonoids delphinidin (2-(3,4,5-Trihydroxyphenyl)chromenylium-3,5,7-triol) and luteolin (2-(3,4-Dihydroxyphenyl)-5,7-dihydroxy-4-chromenone) have been recognized as promising antioxidants and anticancer substances. Due to their extensive use, the goal of the research was to determine whether they have any genotoxic potential in vitro.Methods: Analysis of genotoxic potential was performed applying chromosome aberrations test in human lymphocyte culture, as this kind of research was not conducted abundantly for these two bioflavonoids. Delphinidin and luteolin were dissolved in DMSO and added to cultures in final concentrations of 25, 50 and 100 μM.Results: In human lymphocytes cultures Delphinidin induced PCDs in all treatments, potentially affecting the cell cycle and topoisomerase II activity. In concentration of 50 μM luteolin showed strong genotoxic effects and caused significant reduction of cell proliferation.Conclusion: Luteolin exhibited certain genotoxic and cytostatic potential. Delphinidin was not considered genotoxic, however its impact on mitosis, especially topoisomerase II activity, was revealed

Microsatelite diversity of Bosnian-Herzegovinian-Croatian shepherd dog Tornjak

This paper presents the first estimation of polymorphism of the Bosnian-Herzegovinian-Croatian Shepherd dog Tornjak in Bosnia and Herzegovina using 10 microsatellite loci, which are an integral part of StockMarks® for Canine Genotyping Kit (Applied Biosystems, Foster City, CA, USA). Ten microsatellite loci used in this study are appropriate for assessing the genetic diversity for this breed. Measures of genetic diversity were estimated based on allelic and genotypic calculations, observed (HO) and expected (HE) heterozygosities, deviations from Hardy-Weinberg equilibrium and polymorphism information content (PIC). The lowest genetic diversity was estimated for locus PEZ20, and the highest for PEZ6 locus. Observed and expected mean heterozygosities were 0.7261 and 0.7392, respectively. Statistically significant deviation (p<0.05) from Hardy-Weinberg equilibrium was found for PEZ1, PEZ12, PEZ3 and PEZ6 loci. The PIC values suggested that all markers (100%) are very informative (PIC > 0.5) in terms of their suitability for genetic diversity studies. When all observed parameters are taken into account (observed and expected heterozygosities, PIC, number of detective and effective alleles, number of detected and maximum possible genotypes, major allele frequency and major allele frequency index), we can conclude that PEZ6 locus shows the highest genetic diversity while PEZ3 displays the lowest. However, assuming values of observed and expected heterozygosities, as well as PIC, we consider loci PEZ20 to be the least diverse, but this locus has more effective alleles and more genotypes present than PEZ3. These preliminary results are the first genetic diversity survey of the Bosnian-Herzegovinian-Croatian Shepherd dog Tornjak in Bosnia and Herzegovina and could be useful to the dog breeders in designing and managing breeding strategies. Summarizing the information above, we can conclude that the population of the Bosnian-Herzegovinian-Croatian Shepherd dog Tornjak from B&H is not affected by substantial loss of genetic diversity. Results of our study indicate presence of reasonably high level of genetic variability and lead to a better understanding of this dog breed