AIM:To assess the mucosa-associated bacterial microflora and mucus layer in adolescents with inflammatory bowel disease(IBD) .METHODS:Sixty-one adolescents(mean age 15 years,SD ± 4.13) were included in the study....AIM:To assess the mucosa-associated bacterial microflora and mucus layer in adolescents with inflammatory bowel disease(IBD) .METHODS:Sixty-one adolescents(mean age 15 years,SD ± 4.13) were included in the study.Intestinal biopsies from inflamed and non-inflamed mucosa of IBD patients and from controls with functional abdominal pain were cultured under aerobic and anaerobic conditions.The number of microbes belonging to the same group was calculated per weight of collected tissue.The mucus thickness in frozen samples was measured under a fluorescent microscope.RESULTS:The ratios of different bacterial groups in inflamed and non-inflamed mucosa of IBD patients and controls were specific for particular diseases.Streptococcus spp.were predominant in the inflamed mucosa of Crohn's disease(CD) patients(80% of all bacteria) ,and Lactobacillus spp.were predominant in ulcerative colitis patients(90%) .The differences were statistically significant(P = 0.01-0.001) .Lower number of bifidobacteria was observed in the whole IBD group.A relation was also found between clinical and endoscopic severity and decreased numbers of Lactobacillus and,to a lesser extent,of Streptococcus in biopsies from CD patients.The mucus layer in the inflamed sites was significantly thinner as compared to controls(P = 0.0033) and to non-inflamed areas in IBD patients(P = 0.031) .CONCLUSION:The significantly thinner mucosa of IBD patients showed a predominance of some aerobes specific for particular diseases,their numbers decreased in relation to higher clinical and endoscopic activity of the disease.展开更多
AIM: To determine the features of Enterococcus that contribute to the development and maintenance of the inflammatory process in patients with inflammatory bowel disease (IBD). METHODS: Multiplex polymerase chain reac...AIM: To determine the features of Enterococcus that contribute to the development and maintenance of the inflammatory process in patients with inflammatory bowel disease (IBD). METHODS: Multiplex polymerase chain reaction (PCR) was applied to assess the presence of genes that encode virulence factors [surface aggregating protein (asa1), gelatinase (gelE), cytolysin (cylA), extracellular surface protein (esp) and hyaluronidase (hyl)] in the genomic DNA of 28 strains of Enterococcus isolated from the intestinal tissues of children with IBD (n =16) and of children without IBD (controls; n = 12). Additionally, strains with confirmed presence of the gelE gene were tested by PCR for the presence of quorum sensing genes (fsrA, fsrB, fsrC) that control the gelatinase production. Gelatinase activity was tested on agar plates containing 1.6% gelatin. We also analysed the ability of Enterococcus strains to release and decompose hydrogen peroxide (using Analytical Merckoquant peroxide test strips) and tested their ability to adhere to Caco-2 human gut epithelium cells and form biofilms in vitro. RESULTS: A comparison of the genomes of Enterococcus strains isolated from the inflamed mucosa of patients with IBD with those of the control group showed statistically significant differences in the frequency of theasa1 gene and thegelE gene. Furthermore, the cumulative occurrence of different virulence genes in the genome of a single strain ofEnterococcus isolated from the IBD patient group is greater than in a strain from the control group, although no significant difference was found. Statistically significant differences in the decomposition of hydrogen peroxide and adherence to the Caco-2 epithelial cell line between the strains from the patient group and control group were demonstrated. The results also showed that profuse biofilm production was more frequent amongEnterococcus strains isolated from children with IBD than in control strains. CONCLUSION: Enterococcus strains that adhere strongly to the intestinal epithelium, form biofilms and possess antioxidant defence mechanisms seem to have the greatest influence on the inflammatory process.展开更多
基金Supported by Polish Ministry of Science and Higher Education,Grant No.3PO5E09125
文摘AIM:To assess the mucosa-associated bacterial microflora and mucus layer in adolescents with inflammatory bowel disease(IBD) .METHODS:Sixty-one adolescents(mean age 15 years,SD ± 4.13) were included in the study.Intestinal biopsies from inflamed and non-inflamed mucosa of IBD patients and from controls with functional abdominal pain were cultured under aerobic and anaerobic conditions.The number of microbes belonging to the same group was calculated per weight of collected tissue.The mucus thickness in frozen samples was measured under a fluorescent microscope.RESULTS:The ratios of different bacterial groups in inflamed and non-inflamed mucosa of IBD patients and controls were specific for particular diseases.Streptococcus spp.were predominant in the inflamed mucosa of Crohn's disease(CD) patients(80% of all bacteria) ,and Lactobacillus spp.were predominant in ulcerative colitis patients(90%) .The differences were statistically significant(P = 0.01-0.001) .Lower number of bifidobacteria was observed in the whole IBD group.A relation was also found between clinical and endoscopic severity and decreased numbers of Lactobacillus and,to a lesser extent,of Streptococcus in biopsies from CD patients.The mucus layer in the inflamed sites was significantly thinner as compared to controls(P = 0.0033) and to non-inflamed areas in IBD patients(P = 0.031) .CONCLUSION:The significantly thinner mucosa of IBD patients showed a predominance of some aerobes specific for particular diseases,their numbers decreased in relation to higher clinical and endoscopic activity of the disease.
基金Supported by The Polish Ministry of Science and Higher Education Grants No. 2 PO5A 094 29, 3 P05E 091 25, N N402 0861 and N N401 144638
文摘AIM: To determine the features of Enterococcus that contribute to the development and maintenance of the inflammatory process in patients with inflammatory bowel disease (IBD). METHODS: Multiplex polymerase chain reaction (PCR) was applied to assess the presence of genes that encode virulence factors [surface aggregating protein (asa1), gelatinase (gelE), cytolysin (cylA), extracellular surface protein (esp) and hyaluronidase (hyl)] in the genomic DNA of 28 strains of Enterococcus isolated from the intestinal tissues of children with IBD (n =16) and of children without IBD (controls; n = 12). Additionally, strains with confirmed presence of the gelE gene were tested by PCR for the presence of quorum sensing genes (fsrA, fsrB, fsrC) that control the gelatinase production. Gelatinase activity was tested on agar plates containing 1.6% gelatin. We also analysed the ability of Enterococcus strains to release and decompose hydrogen peroxide (using Analytical Merckoquant peroxide test strips) and tested their ability to adhere to Caco-2 human gut epithelium cells and form biofilms in vitro. RESULTS: A comparison of the genomes of Enterococcus strains isolated from the inflamed mucosa of patients with IBD with those of the control group showed statistically significant differences in the frequency of theasa1 gene and thegelE gene. Furthermore, the cumulative occurrence of different virulence genes in the genome of a single strain ofEnterococcus isolated from the IBD patient group is greater than in a strain from the control group, although no significant difference was found. Statistically significant differences in the decomposition of hydrogen peroxide and adherence to the Caco-2 epithelial cell line between the strains from the patient group and control group were demonstrated. The results also showed that profuse biofilm production was more frequent amongEnterococcus strains isolated from children with IBD than in control strains. CONCLUSION: Enterococcus strains that adhere strongly to the intestinal epithelium, form biofilms and possess antioxidant defence mechanisms seem to have the greatest influence on the inflammatory process.
