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Ulea I.,Cantacuzino National Institute of Research Development for Microbiology and Immunology
Roumanian archives of microbiology and immunology | Year: 2013

Thin layer chromatography (TLC) could easily and rapidly evidentiate the qualitative differences between glycolipids (GLs). Different immunomagnetically purified mycobacterial GLs have been compared using TLC, in order to choose the most appropriate antigens to be utilized in ELISA. The GLs were purified from environmental mycobacteria (EM) (M. avium-intracellulare, M. kansasii, M. xenopi, M. scrofulaceum and M. gordonae) and from M. tuberculosis H37Rv. BioMag Amine and BioMag Carboxyl terminated superparamagnetic microparticles were utilized in the magnetic separation of glycolipids from mycobacterial species. TLC of GLs before and after magnetic purification, corroborated with ELISA results, shows that COOH-terminated particles allow a better purification for M. kansasii, M. xenopi and M. scrofulaceum, while NH2-terminated particles act better on MAI and M. gordonae GLs. The use of GL purified antigens in ELISA could fulfill the criteria of high levels of both sensitivity and specificity of serologic assays in EM diagnosis. Source


Costache A.,Cantacuzino National Institute of Research Development for Microbiology and Immunology
Roumanian archives of microbiology and immunology | Year: 2013

Cyclic AMP (cAMP), one of the most important secondary messengers, is produced by adenylate cyclase (AC) from adenosine triphosphate (ATP). AC is a widespread enzyme, being present both in prokaryotes and eukaryotes. Although they have the same enzymatic activity (ATP cyclization), the structure of these proteins varies, depending on their function and the producing organism. Some pathogenic bacteria utilize these enzymes as toxins which interact with calmodulin (or another eukaryote activator), causing intense cAMP synthesis and disruption of infected cell functions. In contrast, other pathogenic bacteria benefit of augmentation of AC activity for their own function. Based on sequence analysis ofAC catalytic domain from two pathogenic bacteria (Bacillus anthracis and Bordetellapertussis) with known three-dimensional structures, a possible secondary structure for 1-255 amino acid fragment from Pseudomonas aeruginosa AC (with 80TKGFSVKGKSS90 as the ATP binding site) is proposed. Source


Iancu A.D.,Cantacuzino National Institute of Research Development for Microbiology and Immunology
Roumanian archives of microbiology and immunology | Year: 2012

Type 1 diabetes mellitus (T1DM), one of the most prevalent chronic diseases is characterized by the progressive destruction of pancreatic beta cells, leading to insulin deficiency and hyperglycemia. Studies performed on diabetic subjects with prolonged hyperglycemia showed the oxidative stress occurrence followed by molecular, cellular and tissue damage. Currently, reducing the oxidative stress represents a therapeutic target, in order to reduce its complications in diabetic patients. An adequate experimental model of type 1 diabetes represents a prerequisite in oxidative stress study, therefore, we assessed oxidative status in polymorphonuclear cells (PMNs) and peritoneal macrophages using a double transgenic (dTg) mouse model of type 1 diabetes. Our results revealed the increased production of reactive oxygen species (superoxide anion and H2O2) and nitrogen (nitric oxide) species in diabetic mice leading to the idea of oxidative stress model for the study of its complications in type 1 diabetes. Source


Usein C.-R.,Cantacuzino National Institute of Research Development for Microbiology and Immunology | Usein C.-R.,Carol Davila University of Medicine and Pharmacy | Militaru M.,Cantacuzino National Institute of Research Development for Microbiology and Immunology | Cristea V.,Synevo Laboratories | Straut M.,Cantacuzino National Institute of Research Development for Microbiology and Immunology
Memorias do Instituto Oswaldo Cruz | Year: 2014

For the first time, we used multilocus sequence typing (MLST) to understand how Romanian group B streptococcus (GBS) strains fit into the global GBS population structure. Colonising isolates recovered from adult human females were tested for antibiotic resistance, were molecularly serotyped based on the capsular polysaccharide synthesis (cps) gene cluster and further characterised using a set of molecular markers (surface protein genes, pilus-encoded islands and mobile genetic elements inserted in the scpB-lmb intergenic region). Pulsed-field gel electrophoresis was used to complement the MLST clonal distribution pattern of selected strains. Among the 55 strains assigned to six cps types (Ia, Ib, II-V), 18 sequence types (STs) were identified by MLST. Five STs represented new entries to the MLST database. The prevalent STs were ST-1, ST-17, ST-19 and ST-28. Twenty molecular marker profiles were identified. The most common profiles (rib+GBSi1+PI-1, rib+GBSi1+PI-1, PI-2b and alp2/3+PI-1, PI-2a) were associated with the cps III/ST-17 and cps V/ST-1 strains. A cluster of fluoroquinolone-resistant strains was detected among the cps V/ST-19 members; these strains shared alp1 and IS1548 and carried PI-1, PI-2a or both. Our results support the usefulness of implementing an integrated genotyping system at the reference laboratory level to obtain the reliable data required to make comparisons between countries. Source


Iancu A.D.,Cantacuzino National Institute of Research Development for Microbiology and Immunology
Roumanian archives of microbiology and immunology | Year: 2012

Type 1 diabetes is an autoimmune disease affecting a higher and higher number of persons; for this reason, the study of diabetes, and its complications, has shown a major interest. In order to highlight the modifications appeared in this disease, it is essential to use a suitable model. In "Cantacuzino" NIRDMI there is a double transgenic murine model which develops a fulminating form of type 1 diabetes. Previous studies indicate the usefulness of this diabetic murine model in order to study neuropathy. Capsaicin treatment is one method to reduce neuropathic pain. This study was based on the assumption that intraperitoneal administration of a low dose of capsaicin, on a short period of time, can decrease pain sensations generation and transmission. If from the neurological point of view, capsaicin effects are known, its effects on the immune system are not clear yet. Therefore, in this study we have investigated capsaicin effects on oxygen and nitrogen free radicals generation by phagocytic immune cells, in lymphocyte populations, and also capsaicin effects on plasmatic protein oxidation. Our results point to minor modifications in oxygen reactive species production, simultaneous with a significantly decrease in nitric oxide generation, without affecting lymphocyte populations. Therefore, capsaicin short term administration can be used to reduce pain sensations, without the impairment of immune parameters. Source

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