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Opis: Single nucleotide polymorphism (SNP) analysis is important tool in the studies of genetic factors associated with complex diseases and with genetically influenced response to drug therapy (pharmacogenetics). Recently, a new generation of generic dsDNA binding dyes (LCGreen$^{TM}$) contributed to the development of fast and low-cost method for SNP detection and/or genotyping based on high resolution melting (HRM) analysis. The aim of our study was to develop HRM assay for IL23R gene (rs7517847) and to perform association study in Slovenian inflammatory bowel diseases (IBD) patients. We genotyped 345 Slovenian healthy controls and 295 IBD patients including 159 with Crohn's disease (CD) and 136 with ulcerative colitis (UC) for rs7517847 polymorphism in IL23R gene using standard RFLP and optimized HRM methods. In this study, we showed, that HRM is a simple, fast and reliable method for genotyping of clinical samples where homozygotes (GG and TT) were determined by Tm calling method and difference between homozygotes and heterozygotes was determined by different melting curve shape using gene scanning method. With combination of results from Tm calling and gene scanning methods, we achieved 98,6% concordance between PCR-RFLP and PCRHRM results, based on the analysis of 640 samples. We found statistically significant association of IL23R polymorphism with Slovenian Crohn's disease patients when comparing genotype and allele frequencies between CD patients and controls. Allele frequency of minor allele G was 0,46 in controls and was reduced to 0,33 in CD patients (p < 0,001, OR = 0,588). The frequency of T/T genotype carriers was higher in CD patients (50,3%) than in controls (26,7%, p = 0,002, OR = 2,558). We found weak association between IL23R polymorphism and Slovenian UC patients. Carriers of T/T genotype have higher risk for UC (p = 0,035, OR = 1,599). These results suggest IL23R plays important role in CD and UC development in Slovenian patients.
Ključne besede: SNP genotyping, high resolution melting, DNA dyes, inflammatory bowel diseases, LC Green Plus
Objavljeno: 18.08.2017; Ogledov: 56; Prenosov: 0
Polno besedilo (232,19 KB)

Opis: Clostridium difficile is an emerging enteropathogen responsible for pseudomembranous colitis in humans and diarrhoea in several domestic and wild animal species. Despite its known importance, there are few studies aboutC. difficile polymerase chain reaction (PCR) ribotypes in Brazil and the actual knowledge is restricted to studies on human isolates. The aim of the study was therefore to compare C. difficileribotypes isolated from humans and animals in Brazil. Seventy-six C. difficile strains isolated from humans (n = 25), dogs (n = 23), piglets (n = 12), foals (n = 7), calves (n = 7), one cat, and one manned wolf were distributed into 24 different PCR ribotypes. Among toxigenic strains, PCR ribotypes 014/020 and 106 were the most common, accounting for 14 (18.4%) and eight (10.5%) samples, respectively. Fourteen different PCR ribotypes were detected among human isolates, nine of them have also been identified in at least one animal species. PCR ribotype 027 was not detected, whereas 078 were found only in foals. This data suggests a high diversity of PCR ribotypes in humans and animals in Brazil and support the discussion of C. difficile as a zoonotic pathogen.
Ključne besede: Clostridium difficile, pseudomembranous colitis, zoonosis
Objavljeno: 07.08.2017; Ogledov: 36; Prenosov: 0
Polno besedilo (185,91 KB)

Opis: There is no single definition of rural practice available. Definitions vary from one country to another, as countries differ in geography and have different health care systems with varying organizational specificities, even within the same country. In spite of increased urbanization and the specific health-related problems it brings with it, a large proportion of the world population still dwells in rural, remote, and isolated areas. In fact, there are many countries in the world with extensive rural areas. Rural areas are unique in organization, demographics, and infrastructure, and so are the specific health-related problems of people living in them. Healthcare in such areas is generally provided by general practitioners or by physicians specialized in family medicine. One of the basic challenges in rural health is defining which areas are rural and finding the characteristics that define “rural”. There are several criteria and combinations of criteria that can be used to define rural areas. Their use mostly depends on the purpose for which the definition is used, and can thus vary from application to application. This paper addresses issues in rural family practice and criteria that may be used to define such practices. It also presents the use of criteria for defining rural practices in a small European country, on the example of Slovenia.
Ključne besede: Slovenia, rural health, rural population, family practice
Objavljeno: 07.08.2017; Ogledov: 13; Prenosov: 0
Polno besedilo (174,94 KB)

Opis: Disparities in cancer care are a reality of the modern world. Unfortunately, current clinical research is in the hands of for-profit pharmaceutical companies and of researchers from the developed world. Problems specific to cancer care in developing countries and among deprivileged populations are ignored. Independent clinical research can offer new valuable knowledge and identify affordable and cost-effective treatments. As such, research not depending on commercial sponsors should become one of the important avenues to alleviate the problem of cancer disparities.
Ključne besede: clinical research, disparities in cancer care, cost-effective treatment
Objavljeno: 04.08.2017; Ogledov: 43; Prenosov: 0
Polno besedilo (472,46 KB)

Opis: In the recent years, the number of drug- and multi-drug-resistant microbial strains has increased rapidly. Therefore, the need to identify innovative approaches for development of novel anti-infectives and new therapeutic targets is of high priority in global health care. The detection of small RNAs (sRNAs) in bacteria has attracted considerable attention as an emerging class of new gene expression regulators. Several experimental technologies to predict sRNA have been established for the Gram-negative model organism Escherichia coli. In many respects, sRNA screens in this model system have set a blueprint for the global and functional identification of sRNAs for Gram-positive microbes, but the functional role of sRNAs in colonization and pathogenicity for Listeria monocytogenes, Staphylococcus aureus, Streptococcuspyogenes, Enterococcus faecalis and Clostridium difficile is almost completely unknown. Here, we report the current knowledge about the sRNAs of these socioeconomically relevant Gram-positive pathogens, overview the state-of-the-art high-throughput sRNA screening methods and summarize bioinformatics approaches for genome-wide sRNA identification and target prediction. Finally, we discuss the use of modified peptide nucleic acids (PNAs) as a novel tool to inactivate potential sRNA and their applications in rapid and specific detection of pathogenic bacteria.
Ključne besede: anti-infectiveness, peptide nucleic acids, sRNA
Objavljeno: 04.08.2017; Ogledov: 43; Prenosov: 0
Polno besedilo (698,61 KB)