The Frequency of the 677C>T and 1298A>C Polymorphisms in the Methylenetetrahydrofolate Reductase (MTHFR) Gene in the Population.

BACKGROUND: The gene for 5,10-methylenetetrahydrofolate reductase (NAD(P)H) or MTHFR gene encodes protein methylenetetrahydrofolate reductase (MTHFR), an enzyme important in folate metabolism. AIM: The aim of this study was to determine the frequencies of 677C>T and 1298A>C polymorphisms in the MTHFR gene of healthy subjects from the population. MATERIAL AND METHODS: The blood samples were collected from 164 unrelated and healthy donors from population consisted of 98 females and 66 males. Both the MTHFR 677C>T and 1298A>C single nucleotide polymorphisms (SNPs) were analyzed by Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Linkage disequilibrium (LD) between pair of SNPs was calculated through Haploview analysis. RESULTS: The frequency of MTHFR 677T allele in the population (32.62%) was in agreement with the frequency of this allele in most other populations, however, the frequency of MTHFR 1298C allele (38.41%) was higher than that reported for most other populations in the world. Haploview analysis showed a relatively strong LD between 677C>T and 1298A>C SNPs with D' values of 0.87. CONCLUSION: Regarding the two MTHFR polymorphisms, three of the nine combined genotypes were present in 87.2% of the population. 33.54% subjects were complex heterozygous (677CT/1298AC genotype), 34.15% subjects had 677CC/1298AC and 19.51% of 677CT/1298AA genotype. The subjects with 677TT genotype had a 1298AA or 1298AC genotype while subjects with 1298CC genotype had only 677CC genotype. The subjects with 677CC/1298AA genotype were only 3.05%. We were not found triple 677CT/1298CC and quadruple 677TT/1298CC mutation suggesting decreased viability of embryos with increased numbers of mutant alleles.

Cytogenetic Abnormalities Found in Patients with Reproductive Problems.

INTRODUCTION: One of the important causes of male infertility is aberration at the chromosomes. AIM: The main purpose of this study was to determine the frequency and types of chromosomal aberration in infertile/sterile men whose samples were analyzed in the Center for Cytogenetics of Faculty of Medicine University of Sarajevo in the last four years. METHODS: A total of 353 infertile/sterile men, between the ages of 22-55 years, referred for cytogenetic analysis to the Center for Genetics of Faculty of Medicine during the period 2013-2016. Karyotyping was performed on peripheral blood lymphocytes by using the Giemsa trypsin banding (GTG) technique. RESULTS: The structural and numerical chromosomal aberration in infertility/ sterility of men found with the incidence of 6% (20/353). Out of the 20 patients with abnormal cytogenetic diagnosis, structural chromosome abnormalities were observed in 17 (85%) patients and 3 (15%) with numerical aberrations. The type of aberrations mostly found were Robertsonian and reciprocal translocations (35%, 35%, respectively). CONCLUSIONS: The incidence of chromosomal abnormalities in infertile/sterile males suggests that the cytogenetics analysis is an important in male infertility, especially if it will be used for the purpose of assisted reproduction techniques.

Ten-year trends in prevalence of Down syndrome in a developing country: impact of the maternal age and prenatal screening.

OBJECTIVE: This study examines trends in total and live birth prevalence of trisomy 21 (T21) with regard to increasing maternal age and the introduction of prenatal diagnosis in Bosnia and Herzegovina. METHOD: The prenatal detection was introduced in January 2008 in 3 hospitals and assessed until December 31, 2015. In this study, 99 fetuses and 330 babies were diagnosed with T21 in the studied period. RESULTS: On average, each year 33 T21 individuals were born and 13 T21 fetuses were diagnosed prenatally. The calculated incidence for the live born T21 individuals in Bosnia is 1:999. The live-birth prevalence of T21 was 9.6 per 10,000 births and the total prevalence of T21 was 19.1. The total T21 prevalence increases exponentially with the advanced maternal age. Prenatal T21 prevalence is 1.29 per 10,000 births for mothers <35, but increases exponentially with increasing age (32 for >40 years). The most common indications for invasive prenatal testing were ultrasound screening combined with biochemical serum analysis followed by the advanced maternal age. CONCLUSION: The prevalence of liveborn Down syndrome children remained constant. Despite the fact that increasing maternal age in the last decade contributed to the rise in the total T21 prevalence, the effect of the introduction of prenatal diagnosis on the live-birth T21 prevalence of T21 was minimal, leading to the conclusion that the prenatal screening has to be improved in developing countries.

Association of Parental Age and the Type of Down Syndrome on the Territory of Bosnia and Herzegovina.

BACKGROUND: Advanced paternal and/or maternal age is a classic risk factor for Down syndrome. The aim of the study was to investigate the frequency of Down syndrome types in children and its association with maternal and paternal age in Bosnia and Herzegovina. SUBJECTS AND METHODS: The cross sectional, observational study included 127 children, 49 girls and 78 boys, aged 1-180 months suspected to have Down syndrome, admitted to the Centre for Genetics, Faculty of Medicine University of Sarajevo, for cytogenetic analysis and differential diagnosis of Down syndrome during the period from January 2010 to May 2015. Standard method of 72 hours cultivation of peripheral blood lymphocytes has been applied. The accepted level of statistical significance was p<0.05. STUDY RESULTS: The most common type of Down syndrome was standard trisomy (86.6%), comparing to translocation and mosaicism (7.1%; 6.3%, respectively). The highest frequency of Down syndrome cases was in mother and father's group from 30-39 years old (57; 57 children, respectively) compared to mother and father's groups with younger than 30 (44; 29, respectively) and 40 and older (26; 41, respectively). The significant difference was found in maternal age between translocation and mosaicism groups (p=0.036). Difference between parental years and type of Down syndrome was significant when Standard trisomy 21 and translocation (p=0.045), as well as mosaicism and translocation (p=0.036), were compared. CONCLUSION: The most common type of Down syndrome was standard trisomy 21, with highest occurrence in parents from 30 to 39 years old. Parents were the youngest in translocation group. Obtained results suggest that multidisciplinary approach to identifying the trigger for trisomy appearance and the influence of maternal age is required.

Renalase Gene rs2576178 Polymorphism in Hemodialysis Patients: Study in Bosnia and Herzegovina.

INTRODUCTION: Renalase is a protein secreted in kidneys and considered as a blood pressure modulator. High rates of hypertension and its regulation in patients on hemodialysis demands search for potential cause and treatment. The aim of this study was to determine the genotype and allele frequencies of renalase gene rs2576178 polymorphism in population from Bosnia and Herzegovina. Also, the objective of present study was to find the possible association between renalase gene rs2576178 polymorphism and hypertension in patients on hemodialysis. MATERIAL AND METHODS: The genotype of renalase gene rs2576178 polymorphism was determined in 137 participants (100 patients on hemodialysis and 37 controls), using polymerase chain reaction (PCR) and subsequent cleavage with MspI restriction endonuclease. Genotype and allele frequencies were assessed for Hardy-Weinberg equilibrium using a Chi-squared test. The value of P<0.05 was considered as statistically significant. RESULTS: Comparison of genotype distribution and allele frequency in participants on hemodialysis with and without hypertension, and healthy control showed no statistical difference. CONCLUSION: The results of the study suggest that renalase gene rs2576178 polymorphism is not a factor that influences blood pressure in patients on hemodialysis.

Frequency of CCR5Δ32 allele in healthy Bosniak population.

Recent evidence has demonstrated the role of CCR5Δ32 in a variety of human diseases: from infectious and inflammatory diseases to cancer. Several studies have confirmed that genetic variants in chemokine receptor CCR5 gene are correlated with susceptibility and resistance to HIV infection. A 32-nucleotide deletion within the CCR5 reading frame is associated with decreased susceptibility to HIV acquisition and a slower progression to AIDS. Mean frequency of CCR5Δ32 allele in Europe is approximately 10%. The highest allele frequency is observed among Nordic populations (about 12%) and lower in the regions of Southeast Mediterranean (about 5%). Although the frequency of CCR5Δ32 was determined in numerous European populations, there is a lack of studies on this variant in the Bosnia and Hercegovina population. Therefore, the aim of our study was to assess the frequency of CCR5Δ32 allele in the cohort of Bosniaks and compare the results with European reports. CCR5Δ32 was detected by sequence-specific PCR in a sample of 100 healthy subjects from Bosnia and Herzegovina (DNA collected 2011-2013). Mean age of the cohort being 58.8 (± 10.7) years, with 82% of women. We identified 17 heterozygotes and one mutant homozygote in study group, with mean ∆32 allele frequency of 9.5%. CCR5∆32 allele frequency among Bosniaks is comparable to that found in Caucasian populations and follows the pattern of the north-southern gradient observed for Europe. Further studies on larger cohorts with adequate female-to-male ratio are necessary.

Analysis of chromosome aberrations contained in vitro human peripheral blood lymphocytes after treatment with ceftriaxone.

INTRODUCTION: The invention and use of antibiotics in treating infections is one of the greatest achievements of the twentieth century medicine. Antibiotics are one of the categories of pharmaceuticals with a broad and increasing application. GOAL: The goal of this paper is to analyze the influence of different set of test concentrations of ceftriaxone antibiotics on the occurrence of chromosome aberrations after in vitro treatment with concentrations: 0.15 mg/ml, 0.25 mg/ml and 0.50 mg/ml. MATERIALS AND METHODS: In the study was used the blood of healthy donors in vitro, treated with different concentrations of ceftriaxone. Ceftriaxone is a semisynthetic cephalosporine third generation antibiotic, with broad spectrum of activity and with specific characteristics (Nerlove et al. 1996). As a biomarker of genetic damage are used the methods of cultivation of peripheral blood lymphocytes and analysis of chromosome aberrations. Cytogenetic analysis of ceftriaxone genotoxicity was performed in 48-hour culture of human peripheral blood lymphocytes by test of standard chromosome aberration analysis according to Moorhead, with certain modifications. Insight into the frequency and type of chromosome aberrations is obtained by analyzing 100 metaphases per sample. RESULTS: By the analysis of 100 metaphases per sample was determined that the relative frequency of metaphases with chromosome aberrations is increased with increasing concentrations of ceftriaxone in lymphocyte culture. The increase in the frequency of structural aberrations was also positively correlated with the applied ceftriaxone concentrations. Metaphases with numerical and structural aberrations are registered in lymphocyte cultures treated with ceftriaxone concentration of 0.25 mg/ml and 0.50 mg/ml, but this increase was not significant compared to the control cells cultures. CONCLUSIONS: Significantly increased frequency of metaphases with structural chromosome aberrations in cultured human peripheral blood lymphocytes treated with concentrations of 0.25 mg/ml and 0.5 mg/ml compared to the control confirming clastogenic potential of this drug. Ceftriaxone also expressed aneugenic activity at the highest test concentration (0.50 mg/ml), confirming a statistically significant difference in the frequency of numerical aberrations in cultures treated with doses of 0.5 mg/ml.