Infectious Diseases Clinical Hospital
Infectious Diseases Clinical Hospital
Zamolodchikova T.S.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry |
Scherbakov I.T.,Gabrichevsky Scientific Research Institute of Epidemiology and Microbiology |
Khrennikov B.N.,Infectious Diseases Clinical Hospital |
Svirshchevskaya E.V.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry
Biochemistry (Moscow) | Year: 2013
A duodenase, a protease structurally related to human cathepsin G, was found earlier in bovine duodenal mucosa. It was demonstrated that under the influence of duodenase an enteropeptidase zymogen is activated in vitro showing the possible participation of duodenase in the cascade of activation of digestive enzymes. To identify a duodenase functional analog in human duodenum, an immunofluorescence study of duodenal mucosa was conducted by confocal microscopy using antibodies to human cathepsin G and to bovine duodenase. The previously unknown place of synthesis and secretion of cathepsin G - Paneth cells located at the bottom of Lieberkuhn crypts - was revealed. Binding of cathepsin G-specific antibodies in a rough endoplasmic reticulum zone and in the cryptal duct was observed. Duodenase-specific immunofluorescence but not that of cathepsin G was found in the epitheliocytes and secretory ducts of Brunner's glands, which are characteristic sites of duodenase biosynthesis in cattle. Binding of CD14-specific antibodies in the Brunner's glands, where the antibodies co-localized with the antibodies to duodenase, was also demonstrated. These data indicate the presence of a protein immunologically similar to duodenase in the human duodenal mucosa. Our study demonstrated the absence of its colocalization with cathepsin G in Brunner's glands. © 2013 Pleiades Publishing, Ltd.
Petrascu M.,University of Medicine and Pharmacy, Cluj-Napoca |
Flonta M.,Infectious Diseases Clinical Hospital |
Almas A.M.,Infectious Diseases Clinical Hospital
Human and Veterinary Medicine | Year: 2011
Background & objectives: Extended spectrum β-lactamases (ESBLs) have emerged as a major threat worldwide with limited treatment options. The genotypes of ESBL producing strains largely remain unknown in Romania; hence the present study was aimed to determine the genetic characteristics of ESBLs in Escherichia coli and Klebsiella pneumoniae isolates from urine specimens. Material and methods: Total 152 consecutive non-duplicate clinical isolates of E. coli (n=70) and K. pneumoniae (n=82) collected between January 2007 and April 2010 were isolated from urine samples in the Laboratory of Infectious Diseases Hospital Cluj-Napoca. The isolates were examined phenotypically for ESBL production. ESBL strains were further typed for the bla TEM/SHV/CTX-M/O XA genes by PCR using specific primers. Results: The TEM gene was detected in 77.63% (118 of 152) of the isolates followed by CTX-M (110 of 152 [73.36%]), OXA (101 of 152 [66.44%]), and SHV (93 of 152 [61.18%]). From 70 E. coli isolates, 50 (71.42%) were positive for CTXM followed by OXA type (47; 67.14%), TEM (40; 57.14%) and SHV (23; 32.86%). From 82 K. pneumoniae isolates, 78 (95.12%) were positive for TEM followed by SHV type (71; 86.58%), CTX-M (60; 73.17%) and OXA (53; 64.63%). Conclusions: The results of this study provide insights into the genetic characteristics of ESBLs among E. coli and K. pneumoniae isolates from urine specimens. The ESBL-positive E. coli isolates investigated here encoded mainly CTX-Ms (71.42%), followed by OXA type (67.14%), TEM-type (57.14%), and SHV-type (32.86%) β-lactamases. The ESBL-positive K. pneumoniae isolates investigated here encoded mainly TEM type (95.12%) followed by SHV type (86.58%), CTX-M (73.17%) and OXA (64.63%).