J Pacha

Publications of J Pacha

• Sequence analysis of proviral DNA of porcine endogenous retroviruses.

  Authors: G Machnik, D Sypniewski, Z Wydmuch, K Cholewa, U Mazurek, T Wilczok, Z Smorag, J Pacha

  Transplantation proceedings. 01/2006; 37(10):4610-4.

  Among all species analyzed, the domestic pig seems to be the most appropriate organ donor for xenotransplantation. Porcine endogenous retroviruses (PERVs) are present in genomes of all pigs and areAmong all species analyzed, the domestic pig seems to be the most appropriate organ donor for xenotransplantation. Porcine endogenous retroviruses (PERVs) are present in genomes of all pigs and are capable of infecting human cells in vitro thus posing a serious threat for xenotransplantation procedures. Despite the abundant distribution of PERVs integrated with porcine genome, the majority of PERV proviral DNA is not capable of expressing viral proteins unless seriously mutated. The aim of the study was to analyze PERV genome for mutations. The study was performed on blood samples from 146 pigs. Long-range polymerase chain reaction (Long-PCR) was performed with primer sets designed within long terminal repeats (LTRs). Long-PCR products of different molecular weights were obtained: 530 bp (33.1% of individuals), 580 bp (76.7%), 933 bp (100%), and 2900 bp (59.8%). Amplimers of 7200 bp were absent in 12.8% of individuals, indicating the lack of intact proviral DNA. Sequence analysis showed that most PERV proviral DNA was significantly mutated, thus suggesting the inability to express functional viral RNA; however, it cannot be ruled out that compensatory recombination processes could occur enabling replication of defective proviruses.

• [Comparative evaluation of bacteria identification from the Acinetobacter genus using a commercially available API 20NE system, PCR and RFLP techniques]

  Authors: Z Wydmuch, M Kepa, R Wojtyczka, D Idzik, J Pacha

  Medycyna doświadczalna i mikrobiologia. 02/2000; 52(3):267-73.

  A study was carried out for identification of 50 Acinetobacter strains isolated from various clinical materials. Using classic methods the following species were identifies: Acinetobacter sp. (68%),A study was carried out for identification of 50 Acinetobacter strains isolated from various clinical materials. Using classic methods the following species were identifies: Acinetobacter sp. (68%), Acinetobacter baumanii (24%) and Acinetobacter lwofii (8%). In all strains the recA gene was found of 435-500 pz size which confirms their belonging to that genus. Amplification products were digested with restriction enzymes Mbol and HinfI (RFLP) and their detection was carried out on agarose gel by electrophoresis methods, owing to that the arrangement of gene fragment characteristic of each strain was obtained. After careful analysis restriction patterns were obtained corresponding to the following genome species: Acinetobacter baumanii (60%), Acinetobacter sp. 3 (28%) and Acinetobacter lwoffii (12%). The methods of molecular biology made possible a more precise classification of the studied strains according to species. Certain strains determined as Acinetobacter sp. by the API 20NE system were found to be Acinetobacter baumanii, Acinetobacter sp. 3 or Acinetobacter lwofii when determined by the PCR/RFLP method.

• [Evaluation of methicillin-resistance in Staphylococcus aureus by the agar disk diffusion method and PCR]

  Authors: D Idzik, R Wojtyczka, M Kepa, Z Wydmuch, J Pacha, S Głab, I Wróblewska, O Skowronek-Ciołek

  Medycyna doświadczalna i mikrobiologia. 02/2000; 52(4):327-32.

  Staphylococcus aureus is a widespread human pathogen. One the most striking characteritics of this bacterium is resistance to methicillin and all beta-lactam antibiotics. The agar disk diffusionStaphylococcus aureus is a widespread human pathogen. One the most striking characteritics of this bacterium is resistance to methicillin and all beta-lactam antibiotics. The agar disk diffusion method is the most widely used in vitro susceptibility test, but recently molecular methods, e.g. Polymerase Chain Reaction, have been also introduced. We compared the detection of methicillin resistant coagulase positive Staphylococcus aureus isolated from clinical materials in Silesian microbiological laboratories by diffusion method and PCR through the detection of nuc and mec A genes. Our results show that PCR used for the detection of mec A gene increases the detection of methicillin-resistant Staphylococcus aureus strains by 10% as compared to the agar disk diffusion method. Among Staphylococcus aureus strains, detected as methicillin-resistant, 17% of organisms showed no presence of mec A gene.