Subject(s)
Mutation , Urea Cycle Disorders, Inborn/genetics , Alleles , Carbon-Carbon Ligases/deficiency , Carbon-Carbon Ligases/genetics , Child , Child, Preschool , DNA Mutational Analysis , Female , Humans , Infant , Infant, Newborn , Male , Neonatal Screening , Republic of Korea/epidemiology , Urea Cycle Disorders, Inborn/epidemiologyABSTRACT
AIMS: Idiopathic nephrotic syndrome is known as a disease of the renal glomerular epithelial cells (podocytes). Recent advances in podocyte biology showed that podocytopathy is the culprit of nephrotic syndrome. To obtain comprehensive information about the response of podocytes to injury, we investigated the gene expression profile of podocytes in response to puromycin aminonucleoside (PAN)-induced injury. METHODS: Differentiated mouse podocyte cell line (MPC5) cells were treated with 25 microg/ml PAN for 24, 48, or 72 h. Gene expression profiles of these cells were analyzed. Real time PCR analysis was used to confirm the findings of microarray. RESULTS: Expression levels of 23 genes (differentially expressed genes, DEGs), including laminin alpha(1) and MMP3, were significantly different between PAN-treated podocytes and untreated cells. Gene ontology of DEGs indicated that their functional categories were cell adhesion, extracellular matrix (ECM) formation, and ECM degradation. Real-time PCR and indirect immunohistochemistry of PAN-treated and untreated podocytes confirmed the differential expression of DEGs. CONCLUSION: Using unbiased global gene expression profiling, we found that podocytes respond to PAN-induced injury by down-regulating the expression of genes involved in cell adhesion and extracellular matrix.
Subject(s)
Podocytes/metabolism , Puromycin Aminonucleoside/administration & dosage , Transcriptional Activation/physiology , Animals , Cell Line , Dose-Response Relationship, Drug , Gene Expression Profiling , Mice , Podocytes/drug effects , Transcriptional Activation/drug effectsSubject(s)
Iduronidase/genetics , Mucopolysaccharidosis I/genetics , Adolescent , Adult , Alleles , Child , Child, Preschool , DNA Mutational Analysis , Humans , Iduronidase/deficiency , Korea , Mutation , PedigreeABSTRACT
We report a Korean case, consistent with a biochemical diagnosis of trifunctional protein (TFP) deficiency, in which molecular diagnosis revealed a novel mutation in the alpha-subunit of TFP and the rare combination of two intergenic region (C/C and G/G) polymorphisms.
Subject(s)
Amino Acid Metabolism, Inborn Errors/diagnosis , Carnitine/analogs & derivatives , Carnitine/blood , Multienzyme Complexes/deficiency , Multienzyme Complexes/metabolism , Amino Acid Metabolism, Inborn Errors/blood , Cytosine , DNA Mutational Analysis , Fatal Outcome , Guanine , Humans , Infant, Newborn , Male , Mitochondrial Trifunctional Protein , Multienzyme Complexes/genetics , Mutation , Protein Isoforms/deficiency , Protein Isoforms/geneticsABSTRACT
AIMS: To assess the importance of tumour necrosis factor alpha (TNF-alpha) promoter polymorphism in relation to infection with the cytotoxin associated gene A (cagA) subtype of Helicobacter pylori within a dyspeptic Korean population. METHODS: Eighty three patients with gastric disease and 113 healthy controls were studied. The DNA from gastric biopsy specimens was analysed by H pylori specific and cagA specific polymerase chain reaction (PCR). To characterise TNF-alpha polymorphism at positions -308 and -238, PCR based restriction fragment length polymorphism analysis was performed. RESULTS: Helicobacter pylori infection was closely correlated with G to A transition at position -308 of the TNF-alpha promoter when compared with healthy controls (odds ratio (OR), 2.912; 95% confidence interval (CI), 1.082 to 7.836; p = 0.034). Although TNF-alpha -308 polymorphism in patients with H pylori was not significantly different from that in patients without H pylori, the -308A polymorphism was strongly associated with H pylori cagA subtype infection when compared with the polymorphism in cagA negative H pylori infection (OR, 8.757; 95% CI, 1.413 to 54.262; p = 0.019) and healthy controls (OR, 3.683; 95% CI, 1.343 to 10.101; p = 0.011). G to A genetic change at position -238 of the TNF-alpha gene was not significantly associated with H pylori cagA subtype infection. In addition, genetic polymorphisms at both sites of the TNF-alpha promoter in patients with H pylori infection did not correlate with the severity of disease. CONCLUSION: TNF-alpha -308A polymorphism was significantly related to infection with the H pylori cagA subtype in Korean patients with gastric disease.
Subject(s)
Antigens, Bacterial , Bacterial Proteins/genetics , Helicobacter Infections/genetics , Helicobacter pylori/genetics , Polymorphism, Genetic , Stomach Diseases/genetics , Tumor Necrosis Factor-alpha/genetics , Bacterial Typing Techniques , Genetic Predisposition to Disease , Genotype , Helicobacter Infections/complications , Helicobacter Infections/microbiology , Helicobacter pylori/classification , Humans , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Promoter Regions, Genetic , Stomach Diseases/microbiology , Stomach Neoplasms/genetics , Stomach Neoplasms/microbiologyABSTRACT
Here we report that lymphocyte functions were down-regulated by cyanobacterial hepatotoxin microcystin. Treatment of three microcystin (MC) isotypes, MC-LR, MC-YR and nodularin, on B6C3F1 mouse splenocytes produced dose-dependent inhibition of in vitro polyclonal antibody response and lymphoproliferation to LPS. ConA-induced lymphoproliferative response was decreased by MC-YR and nodularin, but no significant effect was observed in the MC-LR treatment. Intraperitoneal administration of nodularin into B6C3F1 mice decreased humoral immune responses to sheep red blood cell (sRBC), and the inhibitory effect became severe when hepatic uptake of nodularin was blocked by rifampicin. Each MC 1 microM suppressed phorbol 12-myristate 13-acetate (PMA) plus ionomycin-induced IL-2 mRNA expression in splenocytes and thymocytes, but not in EL-4 mouse thymoma cells. To further characterize the mechanism for the reduced IL-2 mRNA level, IL-2 mRNA stability was measured using RT-PCR. Deprivation of PMA/ionomycin stimuli from activated splenocytes and blockade of new transcription resulted in destabilization of IL-2 mRNA, which was accelerated by MC treatment. These results demonstrated that MC down-regulated lymphocyte functions and the immunosuppression was mediated, at least in part, through decreased IL-2 mRNA stability.
Subject(s)
Interleukin-2/genetics , Peptides, Cyclic/pharmacology , RNA Stability/drug effects , RNA, Messenger/drug effects , T-Lymphocytes/drug effects , Animals , Antibody Formation/drug effects , Bacterial Toxins/chemistry , Bacterial Toxins/pharmacology , Cell Division/drug effects , Cells, Cultured , Dose-Response Relationship, Drug , Down-Regulation , Female , Lipopolysaccharides/pharmacology , Mice , Mice, Inbred C57BL , Mice, Inbred Strains , Microcystins , RNA, Messenger/genetics , RNA, Messenger/metabolism , T-Lymphocytes/cytology , T-Lymphocytes/metabolism , Tetradecanoylphorbol Acetate/pharmacology , Tumor Cells, CulturedABSTRACT
We investigated the potential association of tumor necrosis factor-alpha (TNF-alpha) promoter polymorphisms with cancers. The study included 169 patients with gastric cancer, uterine cervical cancer, colorectal cancer, or renal cell carcinoma and 92 healthy controls. The -308 and -238 polymorphisms in the TNF-alpha promoter were analyzed by PCR-restriction fragment length polymorphism (RFLP). The proportion of individuals carrying the TNF-238A allele was significantly lower in the cancer group than in the control group. The odds ratio for cancer in subjects with the TNF-238A allele was 0.25 (95% CI, 0.10-0.64). No association was found between the -308 polymorphism and cancers. These results suggest that the -238A allele has a protective function against cancers.