Comparison of Proteome Components of Helicobacter pylori Before and After Mouse Passage

The mouse model is alleged to be a useful tool for understanding of pathophysiological roles of Helicobacter pylori in the development of gastric disorders. However, it has been observed that H. pylori strains significantly differed in their fitness in mice and even mouse strains differed in their susceptibilities to a H. pylori strain. Bacterial components of H. pylori which could affect on its fitness in mice have to be elucidated for the establishment of the mouse model for H. pylori infections. In the comparison of colonization ability between two H. pylori Korean isolates, 51 (isolated from a patient with duodenal ulcer) and 52 (isolated from a patient with gastric cancer), 52 could colonize better than 51 on the gastric mucosa of mouse. Proteome components of H. pylori 52, as a good colonizer and H. pylori 51, as a poor one were quantitatively compared each other. Five bacterial proteins including catalase, urease subunit alpha/beta, enolase and ferritin, were up-regulated in 52. In addition, the respective proteome components of the two strains were also compared with their mouse-passaged homologous strains. Seven and five proteins, which included catalase, flagellin A/B in common, were up-regulated in mouse-adapted 51 and 52, respectively. Among the fourteen identified proteins, urease subunit alpha/beta, flagellin A/B, catalase, ferritin, superoxide dismutase and neutrophil-activation protein have been previously known to be necessary to gastric colonization of H. pylori in animal models. The other up-regulated proteins including enolase, elongation factor Tu and fructose-bisphosphate aldolase have been reported to be associated with acid tolerance of H. pylori. These data provide confirmatory evidence for the importance of those proteins in the development of H. pylori-associated gastric disorders.

Expression of Matrix Metalloproteinase-2 and Tissue Inhibitor of Metalloproteinase-2 in Radiation Exposed Small Intestinal Mucosa of the Rat / 대한방사선종양학회지

PURPOSE: The matrix metalloproteinases (MMPs) are a family of enzymes whose main function is the degradation of the extracellular matrix. Several studies have revealed that MMPs and TIMPs are related to the wound healing process and in photoaging caused by ultraviolet irradiation. However, the expressions of MMP and TIMP after irradiation have not, to the best of our knowledge, been studied. This study investigates the expressions of MMP-2 and TIMP-2 in rat intestinal mucosa following irradiation. Material and Methods:The entire abdomen of Sprague-Dawley rats was irradiated using a single dose method. The rats were sacrificed on day 1, 2, 3, 5, 7 and 14 following irradiation. Histopathological observations were made using hematoxilin & eosin staining. The expressions of MMP-2 and TIMP-2 were examined using immunohistochemistry, immunoblotting and ELISA. RESULTS: Radiation induced damage, associated with atrophic villi, and infiltration of inflammatory cells was observed from the first postirradiation day, and severe tissue damage was observed on the second and the third postirradiation days. An increase in mitosis and the number of regenerating crypts, as evidence of regeneration, were most noticeable on the fifth postirradiation day. From the immunohistochemistry, the MMP-2 expression was observed from the first postirradiation day, but was most conspicuous on the third and the fifth postirradiation days. The TIMP-2 expression was most conspicuous on the fifth postirradiation day. From the immunoblotting, the MMP-2 expression was strongly positive on the third postirradiation day, and that of TIMP-2 showed a strong positive response on the fifth postirradiation day. In ELISA tests, the expressions of MMP-2 and TIMP-2 were increased in the postirradiation groups compared to those of the normal controls, and showed a maximum increase on the fifth postirradiation day. These results were statistically significant. CONCLUSION: The expressions of MMP-2 and TIMP-2 were increased in the intestinal mucosa of the rats following irradiation, and these results correlated with the histopathological findings, such as tissue damage and regeneration. Therefore, this study suggests that MMP-2 and TIMP-2 play roles in the mechanisms of radiation-induced damage and regeneration of intestinal mucosa of rats.

The Effect and Factors Affecting on Lipid Status of Valproate Therapy in Children with Epilepsy

PURPOSE: Antiepileptic drugs may alter serum lipid status in epileptic patients. We conducted this study to assess the effect of valproate on serum levels of total cholesterol(TC), triglycerides(TG), low-density lipoprotein(LDL), high-density lipoprotein(HDL), and TC/HDL ratio, and to investigate the factors affecting serum lipid status in children with epilepsy who had been receiving valproate therapy. METHODS: Thirty epileptic children(16 males, 14 females, mean age 7.4+/-3.3 years) were evaluated for serum lipid status at the onset and the 6, 12 and 24 months of valproate therapy, and were analysed changes and potential factors of affecting changes such as sex, body mass index, valproate concentration, in serum lipid levels during valproate therapy. RESULTS: TC were significantly lowered during first 12 months of valproate theraphy (P<0.05). LDL were lowered during first 12 months. HDL and TC/HDL ratio were not changed and TG were increased during valproate theraphy but not reach to statistical significance. TC, TG, LDL, and HDL return to pretreatment levels after 24 months of valproate theraphy. TC, LDL, HDL, and TC/HDL ratio changes were not significantly different by sex and initial body mass index, but TG were significantly increased in group of BMI below 20(P<0.05). LDL levels were significantly decreased correlation to serum valproate concentration(r=-0.2915. P<0.05). CONCLUSION: Our results suggest that valproate therapy would not increase a risk for atherosclerotic disorders in adulthood, but weight gain with a metabolic consequence of obesity would increase risk for atherosclerotic disorders in adulthood.

The Clinical Analysis of Children Who Increase Urine gamma-hydroxybutyric Acid with Seizure Disorder

PURPOSE: This study was performed to analyse urine gamma-hydroxybutyric acid(GHB) in children with seizures, and to investigate the pattern of seizures and neurologic abnormalities in children related with gamma-hydroxybutyric aciduria. METHODS: We reviewed retrospectively medical records of children who admitted to our hospital with seizures between August 1. 2001 and February 28. 2003. We compared urine GHB levels with controls, and also analyzed the clinical features of patients who showed increased urine GHB. RESULTS: The mean urine GHB was 1.7+/-1.6 mmol/mol cr in febrile seizures, 1.8+/-2.5 mmol/mol cr in non-febrile seizures, and 1.8+/-2.0 mmol/mol cr in controls. Compared with control group, there was no significant difference in urine GHB levels(P>0.05). In 8 of 64 children with seizures, GHB levels increased above 2 standard deviation of normal controls. The types of seizure in children who showed increased urine GHB were generalized tonic clonic seizure in 3 patients, complex partial seizure in 2 patients, febrile seizure in 2 patients, and benign Rolandic epilepsy in 1 patient. 3 patients showed neurologic abnormalities, 4 patients showed electroencephalographic abnormalities, and 2 patients of 6 patients who performed brain imaging study showed brain imaging abnormalities. CONCLUSION: Children with gamma-hydroxybutyric aciduria should be suspected succinic semialdehyde dehydrogenase deficiency as a cause of underlying disease.

Quantification of Age-related Reference Values for Urinary Organic Acids in the Healthy Korean Population

PURPOSE: For the accurate diagnosis of organic acidopathies, quantification of urinary organic acid should be done and we should know the normal ranges of each organic acid excreted in the urine. The amount of organic acids excreted in the urine shows wide variability according to ethnic group, diet and age. We have quantified 82 organic acids to make a Korean reference value. METHODS: Organic acid concentrations were quantified with gas chromatography and the individual acids identified with mass spectrometry in urine specimens from members of the healthy Korean population of ages of one day to more than 12 years, subdivided into four age groups : neonatal period(-2 mon), infantile period(-2 year), childhood period(-12 year) and adolescent and adulthood(over 12 years). For isolation of organic acids from urine, we used solvent extraction method with ethylacetate. Derivatization was done with MSTFA(N-methyl-N-trimethylsilylfluoroacetamide). The library and four points quantification curve for the quantification of each organic acid that we used have been developed by Dr. Giudici of Kayser Permanante Metabolic Laboratory, CA., USA. RESULTS: Quantitative ranges and frequency distribution patterns of urinary organic acid excretion are reported, as a basis on which to compare results obtained for patients whose clinical condition suggests that their excretion values may be abnormal. CONCLUSION: The quantitative values we observed, enable the relative significance of different urinary metabolites to be assessed.