Mannose-binding lectin (MBL) deficiency caused by the variability in the MBL2 gene is responsible for the susceptibility to and severity of various infectious and autoimmune diseases. A combination of six single nucleotide polymorphisms (SNPs) has a major impact on MBL levels in circulation. The aim of this study is to design and validate a sensitive and economical method for determining MBL2 haplogenotypes. The SNaPshot assay is designed and optimized to genotype six SNPs (rs1800451, rs1800450, rs5030737, rs7095891, rs7096206, rs11003125) and is validated by comparing results with Sanger sequencing. Additionally, an algorithm for online calculation of haplogenotype combinations from the determined genotypes is developed. Three hundred and twenty-eight DNA samples from healthy individuals from the Czech population are genotyped. Minor allele frequencies (MAFs) in the Czech population are in accordance with those present in the European population. The SNaPshot assay for MBL2 genotyping is a high-throughput, cost-effective technique that can be used in further genetic-association studies or in clinical practice. Moreover, a freely available online application for the calculation of haplogenotypes from SNPs is developed within the scope of this project.

• Publikační typ
• časopisecké články MeSH

We conducted pooled and meta-analyses of the association of the calpain-10 gene (CAPN10) polymorphisms SNP-43, Indel-19 and SNP-63 individually and as haplotypes with type 2 diabetes (T2D) in 3237 patients and 2935 controls of European ancestry. In the pooled analyses, the common SNP-43*G allele was associated with modest but statistically significant increased risk of T2D (odds ratio (OR)=1.11 (95% confidence interval (CI), 1.02-1.20), P=0.01). Two haplotype combinations were associated with increased risk of T2D (1-2-1/1-2-1, OR=1.20 (1.03-1.41), P=0.02; and 1-1-2/1-2-1, OR=1.26 (1.01-1.59), P=0.04) and one with decreased risk (1-1-1/2-2-1, OR=0.86 (0.75-0.99), P=0.03). The meta-analysis also showed a significant effect of the 1-2-1/1-2-1 haplogenotype on risk (OR=1.25 (1.05-1.50), P=0.01). However, there was evidence for heterogeneity with respect to this effect (P=0.06). The heterogeneity appeared to be due to data sets in which the cases were selected from samples used in linkage studies of T2D. Using only the population-based case-control samples removed the heterogeneity (P=0.89) and strengthened the evidence for association with T2D in both the pooled (SNP-43*G, OR=1.19 (1.07-1.32), P=0.001; 1-2-1/1-2-1 haplogenotype, OR=1.46 (1.19-1.78), P=0.0003; 1-1-2/1-2-1 haplogenotype, OR=1.52 (1.12-2.06), P=0.007; and 1-1-1/2-2-1 haplogenotype, OR=0.83 (0.70-0.99), P=0.03) and the meta-analysis (SNP-43*G, OR=1.18 (1.05-1.32), P=0.005; 1-2-1/1-2-1 haplogenotype, OR=1.68 (1.33-2.11), P=0.00001). The pooled and meta-analyses as well as the linkage disequilibrium and haplotype diversity studies suggest a role for genetic variation in CAPN10 affecting risk of T2D in Europeans.

• MeSH
• běloši * genetika   MeSH
• diabetes mellitus 2. typu * genetika   MeSH
• haplotypy genetika   MeSH
• jednonukleotidový polymorfismus * MeSH
• kalpain * genetika   MeSH
• lidé MeSH
• vazebná nerovnováha * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• metaanalýza MeSH
• multicentrická studie MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Úvod: Konzervativní léčba refluxní choroby jícnu (GERD - gastroesophageal reflux disease) je v současnosti opřena o skupinu léků, které nejúčinněji potlačují sekreci kyseliny solné v žaludku, a to tzv. inhibitory protonové pumpy (IPP). I když se nejedná o kauzální léčbu onemocnění, jsou považovány a akceptovány jako léky volby „zlatého standardu“. Cílem retrospektivní studie bylo zmapovat medikace pacientů s GERD v různých stupních onemocnění, analyzovat jejich individuální variabilitu v genu kódujícím enzym cytochrom P450 (CYP2C19) a navrhnout metodiku pro jednoduché stanovení genového profilu pacienta s GERD pro zefektivnění terapeutických postupů. Metody: Do studie bylo zařazeno celkem 276 osob s GERD po chirurgickém zákroku se známou farmakologickou anamnézou - 94 pacientů s neerozivní refluxní chorobou, 121 s refluxní ezofagitidou a 61 s Barrettovým jícnem (BE - Barrett´s esophagus) nebo adenokarcinomem jícnu (EAC - esophageal adenocarcinoma). Stanovení genotypů dvou polymorfizmů genu CYP2C19 (*17 rs12248560 a *2 rs4244285) bylo založeno na principu kvantitativní polymerázové řetězové reakce. Výsledky: Více než 90 % pacientů užívalo IPP (omeprazol/lansoprazol/pantoprazol). Ačkoli ve sledované kohortě předepsaná dávkování IPP odpovídala doporučení pro udržovací terapii, u pacientů nebyla zohledněna míra schopnosti účinnou látku metabolizovat. Bylo zjištěno, že nositelé genotypových kombinací obsahujících variantu CYP2C9*17 determinující fenotyp ultrarychlého metabolizátoru (UM) mají nižší pravděpodobnost souběžného výskytu s variantou CYP2C19*2, která kóduje fenotyp intermediárního nebo „špatného“ metabolizátoru (IM nebo PM), než osoby se standardní funkcí enzymu CYP2C19 (p = 0,001). Frekvence kombinací genotypů (tzv. haplogenotypů) a ve výsledku fenotypů UM/IM/PM byla mezi pacienty s GERD 37,3/16,7/1,4 %. Závěr: U pacientů s GERD by měl být před zahájením farmakoterapie IPP definován jejich genový profil, resp. kombinace variant CYP2C19*17 a *2, a to vzhledem k jejich vysoké frekvenci v populaci a funkčnímu metabolickému významu. Předpokládáme, že by toto opatření mohlo vést ke zvýšení efektivity farmakoterapie, a tím ke zlepšení kvality života pacientů a snad i k prevenci rozvoje závažnějších stavů, jakými jsou BE a EAC. Nestandardní schopnost metabolizovat IPP by mohla být jedním z předpokladů k indikaci chirurgického zákroku u pacientů s GERD.

Introduction: Conservative treatment of gastroesophageal reflux disease (GERD) is currently based on a group of drugs that effectively suppress the secretion of hydrochloric acid in the stomach, so-called proton pump inhibitors (PPI). Although these drugs do not target the cause of the disease, they are considered and accepted as the “gold standard“ for the treatment of this disease. The aim of this retrospective study was to map medication of GERD patients in various phases of the disease, to analyze individual variability in the cytochrome P450 (CYP2C19) gene, and to propose an effective method for the simple determination of the gene in GERD patients. Methods: The study included 276 GERD patients with known pharmacological anamnesis who had undergone surgical treatment. The subjects included 94 patients with non-erosive reflux disease, 121 with reflux esophagitis, and 61 with Barrett´s esophagus (BE) or esophageal adenocarcinoma (EAC). Genotypes of two polymorphisms in the CYP2C19 gene (*17 rs12248560 and *2 rs4244285) were determined using quantitative polymerase chain reaction. Results: More than 90% patients were treated with PPI (omeprazole/lansoprazole/pantoprazole). Although the prescribed PPI dosing in the studied cohort was consistent with recommendations for maintenance therapy, the patients´ability to metabolize the active substance was not considered. Carriers of genotype combinations containing the CYP2C9*17 variant, which determines the ultra-rapid metabolizer (UM) phenotype, were less likely to co-occur with the CYP2C19*2 variant, which determines the intermediate or “poor“ metabolizer (IM or PM) phenotype, than individuals with the standard functioning CYP2C19 enzyme (p = 0.001). The frequencies of combinations of genotypes (haplogenotypes) and resulting UM/IM/PM phenotypes were 37.3/16.7/1.4% in GERD patients. Conclusion: Prior to initiating PPI pharmacotherapy in GERD patients, determination of CYP2C19 haplogenotypes (CYP2C19*17 and *2 variants) should be performed due to frequent occurence of these variants in population and their functional metabolic significance. We presume that this approach will increase the effectiveness of pharmacotherapy, improve patient quality of life, and very likely help prevent/reduce the risk of the development of more serious conditions, such as BE and EAC. The non-standard ability to metabolize PPI in GERD patients may be one of the indicators for surgical intervention.

• MeSH
• cytochrom P450 CYP2C19 genetika   metabolismus   MeSH
• farmakogenetika * MeSH
• gastroezofageální reflux * diagnóza   farmakoterapie   chirurgie   MeSH
• inhibitory protonové pumpy farmakologie   metabolismus   terapeutické užití   MeSH
• interpretace statistických dat MeSH
• lékové interakce MeSH
• lidé MeSH
• polymorfismus genetický MeSH
• polypharmacy MeSH
• retrospektivní studie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

Periodontitis, an inflammatory disease caused by subgingival Gram-negative (G-) bacteria, is linked with loss of the connective tissue and destruction of the alveolar bone. In the regulation of inflammatory response, chemokine receptor 2 (CXCR2), a specific receptor for interleukin-8 and neutrophil chemoattractant, plays an important role. The first aim of this study was to investigate the CXCR2 gene variability in chronic periodontitis (CP) patients and healthy nonperiodontitis controls in the Czech population. The second aim was to find a relation between CXCR2 gene variants and the presence of periodontal bacteria. A total of 500 unrelated subjects participated in this case-control study. 329 CP patients and 171 healthy nonperiodontitis controls were analyzed using polymerase chain reaction techniques for three single-nucleotide polymorphisms (SNPs): +785C/T (rs2230054), +1208T/C (rs1126579), and +1440A/G (rs1126580). A DNA microarray detection kit was used for the investigation of the subgingival bacterial colonization, in a subgroup of CP subjects (N = 162). No significant differences in allele, genotype, haplotype, or haplogenotype frequencies of CXCR2 gene variants between patients with CP and healthy controls (P > 0.05) were determined. Nevertheless, Aggregatibacter actinomycetemcomitans was detected more frequently in men positive for the C allele of the CXCR2 +785C/T polymorphism (61.8% vs. 41.1%, P < 0.05; OR = 2.31, 95% CI = 1.03-5.20) and for the T allele of the CXCR2 +1208C/T variant (61.8% vs. 38.9%, P < 0.05; OR = 2.54, 95% CI = 1.13-5.71). In contrast, no statistically significant associations of CXCR2 variants with seven selected periodontal bacteria were found in women. Although none of the investigated SNPs in the CXCR2 gene was associated with CP, the CXCR2 gene variants can be associated with subgingival colonization of G- bacteria in men with CP in the Czech population.

• MeSH
• Aggregatibacter actinomycetemcomitans patogenita   MeSH
• alely MeSH
• chronická parodontitida genetika   mikrobiologie   MeSH
• dospělí MeSH
• genotyp MeSH
• haplotypy genetika   MeSH
• jednonukleotidový polymorfismus genetika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• receptory interleukinu-8B genetika   MeSH
• studie případů a kontrol MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Úvod a cíl: Při terapii recidivující aftózní stomatitidy (RAS) se doporučuje tzv. Škachova kúra, při které pacienti užívají kyselinu listovou a vitamin B6. Kyselina listová je v organismu využitelná pouze ve formě aktivního folátu (vitamin B9); v metabolické přeměně sehrává důležitou úlohu enzym methylentetrahydrofolát reduktáza (MTHFR), jehož funkčnost je zásadně ovlivněna genovými polymorfismy. Cílem práce bylo ověřit předpoklad, že podání aktivní formy kyseliny listové spolu s vitaminy B6 a D3 může pozitivně ovlivnit četnost výsevu aft i průběh jejich hojení u pacientů s RAS. Metody: V rámci pilotní studie jsme klinicky vyšetřili deset pacientů, u nichž jsme analyzovali haplogenotypy v genu MTHFR. Podmínky pro zařazení pacientů s RAS byly: věk 18–50 let, česká národnost, geneticky podmíněná snížená schopnost metabolizovat kyselinu listovou (špatný metabolizátor – PM nebo intermediární – IM) a nepřítomnost systémového onemocnění (Crohnova choroba, Behçetova choroba, onemocnění ledvin a další nemoci). Farmakoterapie v upraveném designu dvojitě zaslepené zkřížené studie (cross-over design) probíhala od jara 2018 v délce trvání tří měsíců. Pacientům byl ve třífázovém schématu podáván aktivní folát (glukosaminová sůl 5-methylfolátu), vitaminy B6 a D3, po dobu terapie nesměli užívat žádné jiné potravinové doplňky. Vedli si denní záznamy o svém zdravotním stavu a možných rizikových faktorech RAS a byli v měsíčních intervalech klinicky i laboratorně vyšetřeni. Zaznamenali jsme jak subjektivní hodnocení efektu terapie pacienty, tak i získaná laboratorní data, která byla vyhodnocena statistickým softwarem Statistica v.13. Výsledky: Do studie jsme zařadili tři muže a sedm žen s RAS ve věku 28 až 47 let; tři z nich byli podle genového profilu MTHFR definováni jako PM a sedm jako IM kyseliny listové. Podle subjektivního hodnocení jedna pacientka udala zhoršení stavu, dva pacienti nepozorovali během terapie změnu svého zdravotního stavu a sedm osob bylo spokojeno, neboť u nich došlo k méně častému/žádnému výsevu aft a/nebo doba hojení byla významně kratší. Subjektivní hodnocení terapie pozitivně korelovalo s objektivním klinickým nálezem (r = 0,993, p < 0,001). Z laboratorní analýzy vyplynulo, že podání vitaminu D3 signifikantně zvýšilo sérové koncentrace 25-OH D3 z nízkých/limitních na optimální hodnoty. Medián koncentrací před zahájením studie byl 54,2 nmoll-1, těsně před podáním 68,1 nmoll-1 versus měsíc po podání 96,6 nmoll-1 (norma: 50–175 nmoll-1), (p ≤ 0,01). Další sledované parametry, jako koncentrace kyseliny listové v séru, homocysteinu, ALT, AST a krevní obraz, byly u pacientů ve fyziologickém rozmezí před terapií i po ní. Závěr: Navržená modifikace Škachovy vitaminové kúry se zdá být vhodná pro léčbu pacientů s RAS, u kterých je genetická predispozice ke snížené funkci enzymu MTHFR, neboť během našeho pozorování došlo u 70 % pacientů k subjektivnímu i objektivnímu zlepšení stavu.

SUMMARY Introduction, aim: When treating recurrent aphthous stomatitis (RAS), a so-called Skach‘s therapy, in which patients are administered folic acid and vitamin B6, is recommended. The human body can process folic acid only in the form of active folate (vitamin B9); methylenetetrahydrofolate reductase (MTHFR) plays an important role in metabolic transformation, function of this enzyme is fundamentally influenced by gene polymorphisms. The aim of the study was to verify the assumption that administration of the active form of folic acid together with vitamins B6 and D3 can positively affect the frequency of aphthae/ulcers eruption and the course of their healing in patients with RAS. Methods: In the pilot study, we clinically examined ten patients in whom we analyzed haplogenotypes in the MTHFR gene. Conditions for inclusion of patients with RAS were: age 18–50 years, Czech nationality, genetically impaired ability to metabolize folic acid (poor metaboliser – PM or intermediate – IM) and the absence of systemic disease (Crohn‘s disease, Behçet‘s disease, kidney disease and other diseases). Pharmacotherapy in a modified double-crossed blind study (cross-over design) ran from spring 2018 and lasted three months. Patients were treated with active folate (glucosamine salt 5-methylfolate), vitamins B6 and D3 in the three-phase regimen, they were not allowed to use any other food supplements during the therapy. They kept daily records of their health status and possible risk factors for RAS and were examined clinically and in the laboratory at monthly intervals. We recorded both the subjective evaluation of the effect of the therapy by patients and the obtained laboratory data, which were evaluated by statistical software Statistica v.13. Results: The study was comprised of three men and seven women with RAS, aged 28 to 47 years; according to the MTHFR gene profile, three were defined as PM and seven as IM of folic acid. Based on the subjective assessment, one patient reported a deterioration, two patients did not observe a change in their health state during therapy, and seven were satisfied because they had a lower or no aphthae eruption and/or healing time was significantly shorter. Subjective evaluation of therapy positively correlated with objective clinical finding (r = 0.993, P < 0.001). The laboratory analysis showed that administration of vitamin D3 significantly increased serum concentrations of 25-OH D3 from low/limit to optimal values, median of concentrations prior to study was 54.2 nmoll-1, just before administration 68.1 nmoll-1 vs. month after administration 96.6 nmoll-1 (norm: 50–175 nmoll-1), (P ≤ 0.01). The other studied parameters, such as concentration of folic acid in serum, homocysteine, ALT, AST, and blood count, were within the physiological range before and after therapy. Conclusion: The proposed modification of Skach‘s vitamin therapy seems to be appropriate for the treatment of patients suffering from RAS with genetic predisposition for reduced MTHFR function as during our observation, 70% of patients experienced subjective and objective improvement of their state.

• MeSH
• aftózní stomatitida * terapie   MeSH
• dospělí MeSH
• farmakogenetika MeSH
• klinická studie jako téma MeSH
• kyselina listová terapeutické užití   MeSH
• lidé středního věku MeSH
• lidé MeSH
• vitamin D terapeutické užití   MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• práce podpořená grantem MeSH

BACKGROUND: Recurrent aphthous stomatitis is one of the most prevalent oral mucosal immunological diseases. A recent case-control study in the Egyptian population suggested that single nucleotide polymorphism Gly54Asp (rs1800450) of the mannose-binding lectin 2 gene might affect the mannose-binding lectin serum level and recurrent aphthous stomatitis development. The aim of this study was to determine the distribution of six functional mannose-binding lectin 2 gene polymorphisms and analyse their role in recurrent aphthous stomatitis susceptibility in the Czech population. METHODS: The study included 227 subjects; 137 healthy people and 90 patients with recurrent aphthous stomatitis. Six mannose-binding lectin 2 gene polymorphisms (rs11003125, rs7096206, rs7095891, rs5030737, rs1800450, rs1800451) were analysed by the SNaPshot assay method, mannose-binding lectin serum levels were determined by enzyme-linked immunosorbent assay (ELISA) method in a subgroup of subjects (N = 87). RESULTS: No significant differences in mean of mannose-binding lectin serum levels between healthy controls and patients with recurrent aphthous stomatitis were observed (383 ng/ml ± 249 standard deviation (SD) vs. 316 ng/ml ± 177 SD in remission phase vs. 343 ng/ml ± 254 SD in active phase; p > 0.05), also the allele and genotype frequencies of the studied mannose-binding lectin 2 polymorphisms did not differ significantly between the two groups (p > 0.05, odds ratio (OR): 0.75-1.23). Moreover, the distribution of mannose-binding lectin 2 haplotypes and haplogenotypes was similar in the healthy subjects and patients with recurrent aphthous stomatitis (p > 0.05, OR: 0.75-1.23). CONCLUSIONS: This study did not confirm the previously reported association of the mannose-binding lectin 2 Gly54Asp gene variant and low mannose-binding lectin serum level as the risk factors for susceptibility to recurrent aphthous stomatitis. In addition, no significant relationships between mannose-binding lectin 2 functional haplotypes or haplogenotypes and recurrent aphthous stomatitis were observed.

• MeSH
• aftózní stomatitida * genetika   MeSH
• frekvence genu MeSH
• genetická predispozice k nemoci MeSH
• genotyp MeSH
• jednonukleotidový polymorfismus MeSH
• lektin vázající mannosu MeSH
• lidé MeSH
• studie případů a kontrol MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

To this date, there are no recommendations for personalized stress ulcer prophylaxis (SUP) in critical care that would take the patient's individual genetic predispositions into account. Of drugs used for this purpose, proton pump inhibitors (PPIs) are the first-choice drugs in intensive care unit patients. The degradation of proton pump inhibitors is mediated by cytochrome P450 (CYP) enzymes; in particular, CYP2C19 and, to a lesser extent, CYP3A4 are involved. Expression and metabolic activity of, namely in, CYP2C19 is significantly affected by single nucleotide polymorphisms, the drug metabolization rate varies greatly from ultrarapid to poor and likely influences the optimal dosage. As these CYP2C19 predictive phenotypes via CYP2C19 haplogenotypes (rs12248560/rs4244285) can be relatively easily determined using the current standard equipment of hospital laboratories, we prepared a set of recommendations for personalized PPI-based stress ulcer prophylaxis taking into account the patient's CYP2C19 predictive phenotype determined in this way. These recommendations are valid, in particular, for European, American and African populations, because these populations have the high representations of the CYP2C19*17 allele associated with the overexpression of the CYP2C19 gene and ultrarapid degradation of PPIs. We propose the CYP2C19 gene profiling as a tool for personalized SUP with PPI in critically ill patients.

• Publikační typ
• časopisecké články MeSH