XIX, 483 s. : tab., grafy ; 24 cm
Cieľ práce: Cieľom štúdie bolo stanovenie prevalencie ESBL-pozitívnych izolátov Klebsielh pneumoniae u pacientov v intenzívnej starostlivosti a ich molekulárno-biologická analýza. Materiál a metódy: V období 5 mesiacov boli od pacientov hospitalizovaných na Klinike anestéziológie a resuscitácie Fakultnej nemocnice Olomouc izolované kmene Klehsiella pneumoniae. U každého izolátu bol určený antibiogram štandardnou dilučnou mikrometódou a produkcia ESBL bola stanovená modifikovaným double disk synergy testom. Na dôkaz prítomnosti génu blaTEM a blaSHV bola použitá PCR. Izoláty produkujúce SHV a TEM typy β-laktamáz boh ďalej typizované použitím metódy polymorfizmu dĺžky restrikčných fragmentov (RFLP) na identifikáciu najbežnejšie sa vyskytujúcich mutácií zodpovedných za vznik ESBL fenotypu. Posúdenie podobnosti, resp. identity izolátov, bolo uskutočnené pulznou gelovou elektroforézou (PFGE) fragmentov DNA, naštiepených pomocou reštrikčnej endonukleázy Xbal. Výsledky: Celkovo bolo získaných 67 izolátov Klebsiella pneumoniae. U 13 z nich bola stanovená produkcia ESBL a pomocou PCR dokázaná pritomnost WasHv génu. Reštrikčné štiepenie pomocou Nhel odhalilo výskyt mutácie v pozícii 238 u všetkých SHV-pozitívnych PCR produktov. Prítomnosť génu kódujúceho širokospektrú β-laktamázu TEM typu však nebola potvrdená. Molekulárno-biologická typizácia pomocou PFGE zistila prítomnosť 11 rôznych kmeňov. Záver. Prevalencia ESBL-pozitívnych kmeňov KlebsieJJa pneumoniae dosiahla u sledovanej skupiny pacientov v intenzívnej starostlivosti hodnoty 19,4 %. Analýza SHV a TEM produktov PCR metódou RFLP preukázala výskyt ESBL typu SHV. Celkovo 84,6 % kmenov malo jedinečný reštrikčný profil. Výsledky dokladujú nielen dobrú úroveň hygienicko-epidemiologických režimov na sledovanej klinike, ale aj racionálnu antibiotickú politiku.
Objectives: The study aimed at the assessment of the prevalence of ESBL-positive isolates of Klebsiella pneumoniae in intensive care patients and their molecular biology analysis. Material and methods: Over a 5-month period, Klebsiella pneumoniae strains were isolated from patients hospitalized at the Department of Anaesthesiology and Resuscitation of the University Hospital in Olomouc. For each isolate, an antibiogram was performed by the standard microdilution method and the production of ESBL was determined by the modified double-disk synergy test. PCR was used to demonstrate the presence of the blaTEM and blaSHV genes. The isolates producing SHV- and TEM-types of β-lactamases were typed using the restriction fragment length polymorphism (RFLP) method to identify the most common mutations responsible for the development of an ESBL phenotype. Similar or identical isolates were determined by pulsed-field gel electrophoresis (PFGE) of DNA fragments cleaved by the XbaI restriction endonuclease. Results: A total of 67 isolates of KJebsieJJa pneumoniae were obtained. In 13 of them, the production of ESBL was detected and the presence of the blaSHV gene was confirmed by PCR. Restriction cleavage by Nhei revealed mutations at position 238 in all SHV-positive PCR products. The restriction analysis did not confirm the presence of the gene encoding TEM-type extended-spectrum β-lactamase. Molecular biology typing by PFGE detected the presence of 11 different strains. Conclusions: In the observed group of intensive care patients, the prevalence of ESBL-positive strains of Klebsiella pneumoniae reached 19.4 %. The analysis of SHV and TEM products of PCR by the RFLPP method showed the prevalence of SHV-type ESBL. Overail, 84.6 % of the strains had unique restriction profiles. The results suggest both high levels of hygienic and epidemiological measures at the monitored department and rational antibiotic policy.
Východisko. Wilsonova choroba je autozomálně recesivní onemocnění charakteristické akumulací a intoxikací mědi v organizmu. Molekulární podstatou onemocnění jsou mutace v genu pro měď transportující ATPázu (ATP7B). Metody a výsledky. Předmětem sdělení jsou výsledky molekulárně-genetických vyšetření ve 130 nepříbuzných rodinách s výskytem Wilsonovy choroby. Pomocí denaturační gradientové gelové elektroforézy (DGGE) byly detekovány exony s abnormální sekvencí. Následným sekvenováním bylo nalezeno celkem 17 kauzálních mutací a 9 polymorfizmů. Pět mutací bylo dosud nepopsaných. Z celkového počtu 260 mutantních alel bylo 214 (82,3 %) identifikováno. Nejčastější mutace, H1069Q, se v naší populaci vyskytuje s frekvencí 65,8 %. Frekvence ostatních mutací nepřevyšuje 5 %. Závěry. DNA diagnostika Wilsonovy choroby v postižených rodinách nabízí pohotové a spolehlivé výsledky. V rámci genetického poradenství umožňuje odhalit asymptomatické a heterozygotní sourozence.
Background. Wilson disease is an autosomal recessive disorder, characterized by cooper accumulation and intoxication of the organism. Molecular basis of the disease represent mutations in the gene for the copper-transporting ATPase (ATP7B). Methods and Results. The submitted paper deals with results of molecular-genetic examination in 130 unrelated families in whichWilson disease was diagnosed. By denaturing gradient gel electrophoresis (DGGE), the exons with abnormal sequences were detected. Followed by sequencing, 17 causal mutations and 9 silent polymorphism were found. Five novel mutations were detected. After analysis of 260 mutant alleles, 214 (82.3 %) were identified. The most frequent mutation, H1069Q, occurred in our population with the frequency of 65.8 %. Incidence of other mutations, however, did not exceed 5 %. Conclusions. DNA analysis of the Wilson disease offers prompt and reliable results in affected families. It can help to identify asymptomatic and heterozygote siblings at genetic counselling.
- MeSH
- Adenosine Triphosphatases MeSH
- Child MeSH
- Adult MeSH
- Research Support as Topic MeSH
- Genes MeSH
- Hepatolenticular Degeneration diagnosis etiology MeSH
- Humans MeSH
- Copper metabolism urine MeSH
- Molecular Biology methods MeSH
- Mutation MeSH
- DNA Mutational Analysis MeSH
- Check Tag
- Child MeSH
- Adult MeSH
- Humans MeSH
- Publication type
- Review MeSH
- Comparative Study MeSH
A simple molecular modeling method for the characterization of polymeric drug carriers is presented. Six biodegradable polymers have been investigated as drug carriers using molecular simulations: l-polylactide, d-polylactide, chitosan, polyglycolic acid, polyethylene glycol and cellulose. Cyclosporine A has been chosen as a model drug substance. Classical molecular dynamics and docking calculations were employed to model and predict polymer-drug interactions. These interactions have been analyzed by non-bond interaction energy and interaction parameter calculated using Flory-Huggins theory. Flexibility of polymer chains has been characterized by the change of gyration radius along the molecular dynamics trajectory. The relationship between mixing energy, chain length and chain flexibility has been revealed for each polymer/drug system.
We have identified seven different beta-thalassemia mutations and one delta beta-thalassemia determinant (the Sicilian type) in 32 members of 17 Hungarian families. The most common mutation is the IVS-I-1 (G-->A) change; its high frequency is comparable to that observed in neighboring Czechoslovakia. Additional mutations are of Mediterranean origin. One rare mutation (initiation codon ATG-->GTG) was identified as an independent mutation because of the absence of known polymorphisms in the beta-globin gene. One new frameshift at codon 51 (-C) was observed in a single individual; hematological data were as expected for a beta zero-thalassemia heterozygosity.
- MeSH
- beta-Thalassemia epidemiology genetics MeSH
- DNA analysis MeSH
- Globins genetics MeSH
- Codon MeSH
- Humans MeSH
- Molecular Biology MeSH
- Molecular Sequence Data MeSH
- Oligonucleotide Probes MeSH
- Frameshift Mutation genetics MeSH
- Amino Acid Sequence MeSH
- Base Sequence MeSH
- Check Tag
- Humans MeSH
- Publication type
- Research Support, U.S. Gov't, P.H.S. MeSH
- Geographicals
- Hungary MeSH
Plum pox virus (PPV), the agent responsible for sharka disease, is the most important viral pathogen of stone fruit trees. The fruits of these fruit species are widely used in the processing industry, thus being economically very attractive. This viral disease significantly reduces the vitality of the fruit trees and the quantity and quality of fruits. The present review describes recent methods used for the identification and characterization of economically important Plum pox virus. Understanding the diversity of plant viruses is an essential step to design efficient management strategies to eliminate economical losses.
