Association of rs211037 GABRG2 gene polymorphism with susceptibility to idiopathic generalized epilepsy

Aim This case-control study aimed to determine a possible association of single nucleotide polymorphism rs211037 of the gamma-aminobutyric acid receptor subunit gamma-2 (GABRG2) gene with the susceptibility to idiopathic generalized epilepsy (IGE) in the Macedonian population. Methods It enrolled 96 patients with clinically verified IGE and 51 healthy individuals without personal and family history of epilepsy or other neurological disorders as controls. A determination of the GABRG2 rs211037 polymorphism was performed using the TaqMan-based genotyping assay. Results A significant dominant association of the CC genotype (odds ratio - OR=2.100, 95% CI=1.018-4.332; p=0.043) and allelic association of C allele (OR=1.902, CI=1.040-3.477; p=0.035) with susceptibility to IGE was found. Carriers of CC genotype had approximately a 2-fold higher probability of developing IGE than the carriers of CT and TT genotypes. Carriers of the C allele had a 1.9-folds higher probability for IGE than the carriers of the T allele. Conclusion The polymorphism rs211037 of the GABRG2 gene increases the risk of the development of idiopathic generalized epilepsy in the Macedonian population

DEEP VEIN THROMBOSIS AND RECURENT PULMONARY EMBOLISM IN A PATIENT WITH THROMBOPHILIC MUTATIONS AND GENERALIZED PSORIASIS: A CASE REPORT

Introduction: Genetic risk factors that increase venous thromboembolism risk are disorders in the synthesis or activity of coagulation factors. Factor V Leiden, prothrombin (20210-A), antithrombin deficiency, protein C and protein S deficiency, and hyperhomocysteinaemia are the most common venous thromboembolism-related gene mutations. When genetic factors are combined with non-provoking risk factors (obesity, psoriasis, smoking and previous venous thromboembolism) the result is increased venous thromboembolism risk for each factor individually. Previous venous thromboembolism is one of the strongest risk factors, even in patients actively treated with anticoagulant. Patients are more likely to have recurrent venous thromboembolism with longer duration. Psoriasis is a complex immune–mediated disease, associated with cardiovascular risk, hypercoagulability markers and elevated homocysteine. Lots of observational reports suggest increased incidence of venous trombembolic events in patient with psoriasis. Case presentation: We present patient with inherited thrombophilia and chronic diffuse plaque psoriasis complicated with deep venous thrombosis and pulmonary embolism. DNA analysis indicates the presence of homozygosis for Factor V Leiden mutation as well as heterozygosis for Factor XIII V34L, PAI -1 5G/4G and MTHFR A1298C polymorphism. Dermatological anamnesis is positive for plaque psoriasis since 12 years ago. Conclusion: The presentation of this case indicates an association between venous thromboembolism and chronic psoriasis. All patients with recurrent thromboembolism, hereditary thrombophilia, and moderate to severe psoriasis should be considered to be at higher risk for venous thromboembolism and appropriately treated

Влијанието на Ц677Т полиморфизмот на генот за МТХФРврз инциденцијата на токÑичните ефекти од виÑоките дози МТХ кај деца Ñо акутна лимфоблаÑтна леукемија

In current protocols for treatment of acute lymphoblastic leukemia in childhood methotrexate (MTX) is one of the crucial cytostatics. The occurrence of MTX toxicity is still  Ð° great problem because of the interpatient differences in drug metabolism. These differences may  be due  to polymorphisms of genes involved in the folate metabolism. The present study was carried out to determine the prevalence of MTHFR  C677T polymorphism in children with acute lymphoblastic leukemia (ALL). Also the effect of the genotype on the toxic  effects during therapy with high doses of МТХ in 45 patients with ALL treated in accordance with the protocol ALL BFM 95 and ALL BFM 2000 was evaluated. All 45 patients with ALL were genotyped for MTHFR C677T polymorphism. Correlation with the presence of a certain polymorphism and toxic effects of the chemotherapy with high doses of МТХ was  made in the Department  for hematology and oncology at the University Clinic for children's diseases – Skopje. The control group included 32 healthy patients. In the study group 24 (53.3%) children had a wild type of polymorphism (CC), 15 (33.33%) children were heterozygous (CT) for MTHFR C677T polymorphism, and 6 (13.33%) children were homozygous for the variant type  of the polymorphism (ТТ). The  correlation of the genotype with МТХ toxicity indicated a statistical significance only for oral mucositis, while for the other toxic effects there was no statistically significant correlation. In our study the results indicated that oral mucositis was statistically significantly more frequently identified in the case of the variant carriers for this polymorphism. For the other toxic effects caused by the therapy with high doses of МТХ no statistically  significant correlation with MTHFR C677T polymorphism was  identified. This  occurrence maybe due  to the small number of patients analyzed in this study and the possible protective influence of other genetic polymorphisms included in the folate metabolism, which were not subject to consideration of this study.Во актуелните протоколи за третман на акутна лимфоблаÑтна леукемија во  Ð´ÐµÑ‚Ñката возраÑÑ‚ метотрекÑатот (МТХ) е еден од круцијалните цитоÑтатици. Предвидувањето на појавата на токÑични ефекти во тек на терапијата Ñо МТХ претÑтавува Ñè уште голем проблем заради индивидуалните разлики во метаболизмот на лекови кај пациентите. Овие разлики можеби Ñе должат на приÑуÑтво на полиморфизми на гените вклучени  Ð²Ð¾  Ñ„олатниот метаболизам. Целта на  Ñтудијата беше  Ð´Ð°  Ñе одреди инциденцијата на МТХФР Ц677Т полиморфизмот кај децата Ñо акутна лимфоблаÑтна леукемија (ÐЛЛ) и да Ñе анализира влијанието на генотипот врз  Ð¼Ð°Ð½Ð¸Ñ„еÑтацијата на токÑичните ефекти во тек на терапија Ñо виÑоки дози МТХ кај пациенти Ñо ÐЛЛ третирани по  Ð¿Ñ€Ð¾Ñ‚околот ÐЛЛ БФМ  95. Беше вклучена и контролна група  Ð¾Ð´ 32 здрави иÑпитаници. Беше  Ð½Ð°Ð¿Ñ€Ð°Ð²ÐµÐ½Ð° генотипизација за полиморфизмот Ц677Т кај 45 пациенти Ñо ÐЛЛ и негова корелација Ñо токÑичните ефекти од хемотерапијата Ñо виÑоки дози  ÐœÐ¢Ð¥ анализирани од болничките иÑтории на пациенти Ñо ÐЛЛ лекувани на Одделот за хематологија и онкологија при  ÐˆÐ—У УниверзитетÑка клиника за детÑки болеÑти - Скопје.  Ð’о ÑтудиÑката група 15 (33,33%) пациенти беа хетерозиготи (ЦТ) за полиморфизмот Ц677Т на генот MTHFR, а 6 (13,33%) пациенти хомозиготи (ТТ). Во контролната група  10 иÑпитаници  (31,25%) беа  хетерозиготи за полиморфизмот (ЦТ), а 6 иÑпитаници (18,75%) хомозиготи (ТТ). Корелацијата на генотипот Ñо токÑичните ефекти од виÑоките дози  Ð½Ð° МТХ покажа ÑтатиÑтичка ÑигнификантноÑÑ‚ Ñамо за орален мукозит, додека беше ÑтатиÑтички неÑигнификантна за оÑтанатите токÑични ефекти. Оваа појава можеби Ñе должи на малата група пациенти којашто беше анализирана во оваа Ñтудија и можното протективно влијание на  Ð´Ñ€ÑƒÐ³Ð¸  Ð³ÐµÐ½Ñки полиморфизми вклучени во фолатниот метаболизам, а кои не беа предмет на оваа Ñтудија

Optimized genotyping method for identification of bacterial contaminants in pharmaceutical industry

Microbiological control has crucial importance in the pharmaceutical industry regarding the possible bacterial contamination of the environment, water, raw materials and finished products. Molecular identification of bacterial contaminants based on DNA sequencing of the hypervariable 16S rRNA gene has been introduced recently. The aim of this study is to investigate the suitability of gene sequencing using our selection of PCR primers and conditions for rapid and accurate bacterial identification in the pharmaceutical industry quality control. DNA was extracted from overnight incubated colonies from 10 bacterial ATCC strains, which are common contaminants in the pharmaceutical industry. A region of bacterial 16S rRNA gene was analyzed by bidirectional DNA sequencing. Bacterial identification based on partial sequencing of the 16S rRNA gene is the appropriate method that could be used in the pharmaceutical industry after adequate validations. We have successfully identified all tested bacteria with more than 99 % similarity to the already published sequences

Human Papillomavirus infection in cervical precancerous lesions

Infection with high-risk HPV genotypes increases the risk for persistent or progressive cervical lesions. The aim of this study was to determine the frequency and correlation of the HPV infection with the cytological or histopathological diagnosis. A total of 6988 samples were analysed and after exclusion of the repeated samples and/or those containing degraded DNA, 4421 patients were included in the study group. PCR-RFLP technique was used as a method for HPV typing. HPV infection was detected in 1819 out of 4421 patients. The frequency of infection was 23% in the patients’ group with cytological and 60% in the group with histopathological diagnosis. The HPV frequencies in precursor lesions were as follows: 51% in patients with mild; 75% in patients with moderate; 91% in patients with severe dysplasia and 93% in carcinoma in situ lesions. The most prevalent HPV types in descending order were: HPV 16, 31, 53, etc. HPV infection was the most frequent in patients under 19 years old. In 169 out of 1253 samples with determined viral genotype, a multiple infections were found. This data for the prevalence and distribution of the HPV infection in Macedonian women accentuates the need for the establishment of organized screening programs

HPV status after cold knife conization

Introduction. The aim of this study was to determine whether HPV DNA test after cold knife conization is a predictive factor for CIN persistence or recurrence. The study also investigated whether HPV DNA test results should influence post cold knife excision surveillance. Materials and methods. A retrospective observation study was performed on 738 patients who underwent cold knife conization for CIN or microinvasive cervical cancer at the University Clinic of Obstetrics & Gynecology, Medical Faculty, Ss. Cyril and Methodius University, Skopje from 1st June 2007 to 1st June 2009. A total of 217 patients met the inclusion criteria and were with complete data. The follow-up HPV DNA testing was performed at 8 months after cold knife conization, after which the patients were followed-up every 4 months till 24 months postoperatively. Results. HPV DNA testing after 8 months after conization showed that 44 patients were HPV DNA positive and 199 were HPV DNA negative. Recurrent cytological abnormalities were found in 26 of the 44 HPV DNA positive patients, and in 12 of the 199 HPV DNA negative patients. Analysis showed that a positive HPV DNA result was a risk factor for recurrent/persistent cervical intraepithelial neoplasia. Conclusion. HPV DNA testing 8 months after conization is important for predicting the risk of disease: persistence or recurrence. In addition, such testing can assist in designing patient management, since HPV DNA negative patients should undergo routine surveillance, while HPV DNA positive patients should undergo frequent and meticulous surveillance

Comparative analysis of microsatellite dna in well sarhplaninian shepherd and well selected sarhplaninian shepherd

DNA microsatellites are codominant genetic markers and they are widely used in the characterization of biodiversity. Genetic variability of sarplaninian shepherds populations was determinate on the basis of analyses of ten DNA microsatellites loci (FH2361, DGN10, FH3287, FH3924, FH3608, FH3023, FH3489, FH3721, FH4027 and FH2141). In this study we have included 37 representative individuals from sarplaninian shepherd (19 lowland sarplaninians and 18 mountain sarplaninians). Informativnes of DNA microsatellites loci was determinate with parameter (Polymorphism Informative Content- PIC), and according to the determinate number of alleles for each locus. All ten DNA microsatellite loci were highly polymorphic. In the genome of sarplaninian shepherd loci FH2141 showed highest PIC value (0.975), and the loci with lowest PIC value was FH3924 locus (0.812). Number of detected alleles for each locus in the genome of sarplaninian shepherd varies from 16 (FH2361) to 33 (FH3023 and FH3489). Intrapopulation genetic variability was determinate according to the number alleles for each locus separately in each population, mean number of alleles for all eight loci and with number of the characteristic alleles. The most common alleles in populations of sarplaninian shepherds were detected in FH2141 loci, 49 (35 heterozygous and 14 homozygous). Taking in consideration the research results from this study a conclusion can be drawn that the genetic variability in Sarplaninian shepherd population is very low. This conclusion and similar stand points in the studies from a significant number of other authors, implicates that the Sarplaninian shepherd is relatively pure breed rarely interbred with phonotypic similar dogs in order to improve the characteristics of the breed

Overexpression of UHRF1 gene correlates with the major clinicopathological parameters in urinary bladder cancer

ABSTRACT Introduction Recently, expression of the UHRF1 gene was found to be up-regulated in numerous neoplasms, including the urinary bladder transitional cell carcinoma (TCC). Objective The aim of our study was to determine if the expression levels of UHRF1 gene correlates with the major pathological characteristics of the tumor and patients’ clinical outcome. Materials and Methods In our study, we have analyzed the tissue samples derived from group of 70 patients with histologically confirmed TCC of the urinary bladder, while normal urinary bladder mucosa obtained from 40 patients with nonmalignant diseases was used as a negative control group. Expression of UHRF1 gene in each patient sample was determined using reverse transcriptase-polymerase chain reaction. Results UHRF1 gene expression was found to be app. 2.5 times higher in samples from patients with TCC in comparison with normal epithelium derived from control group patients. Analysis show that gene expression correlates with the malignancy of the tumor. A highly significant differences were found between the expression values of samples from low and high grade TCC, as well as between the high grade and control group. UHRF1 expression was higher in patients with non-muscle invasive disease than in those with muscle invasive disease. Conclusions The result of this study indicates that UHRF1 gene expression levels correlates with the major pathological characteristics of TCC samples and with the clinical outcome of those patients. Determination of UHRF1 gene expression could have a potential to be used as a sensitive molecular marker in patients with urinary bladder cancer

Optimized genotyping method for identification of bacterial contaminants in pharmaceutical industry

Microbiological control is of crucial importance in the pharmaceutical industry regarding the possible bacterial contamination of the environment, water, raw materials and finished products. Molecular identification of bacterial contaminants based on DNA sequencing of the hypervariable 16SrRNA gene has been introduced recently. The aim of this study is to investigate the suitability of gene sequencing using our selection of PCR primers and conditions for rapid and accurate bacterial identification in pharmaceutical industry quality control

High frequency of the HRAS oncogene codon 12 mutation in Macedonian patients with urinary bladder cancer

Point mutations at codon 12 of the HRAS (v-Ha-ras Harvey rat sarcoma viral oncogene homolog) oncogene are one of the best defined and widely studied molecular genetic events in transitional cell carcinoma (TCC) of the urinary bladder. The aim of this study was to use the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of paraffin-embedded tissue-derived DNA to determine the frequency of the HRAS oncogene G ->T codon 12 mutation in TCC patients being treated at the University Urology Clinic in Skopje, Republic of Macedonia. DNA isolated from paraffin-embedded tissue (PET) surgically removed TCC specimens of 62 (81.58%) out of 76 patients were successfully amplified, the remaining 14 (18.42%) showing compromised DNA integrity. The codon 12 mutation of the HRAS oncogene was found in 24 (38.71%) out of 62 successfully tested TCC urinary bladder samples. No significant relationship between the mutation frequency and the histopathological grade of tumor differentiation was detected (chi² = 0.044; p = 0.978). The relatively high frequency of mutations found in our study was comparable with some of the previously reported data obtained by this and/or other PCR-based methods. This highly sensitive and specific PCR-RFLP analysis was demonstrated to be a suitable method for the detection of mutations at codon 12 of the HRAS oncogene in PET samples of urinary bladder TCC