Rhomboid family member 2 gene (Rhbdf2) is an inactive homologue lacking essential catalytic residues of rhomboid intramembrane serine proteases. The protein is necessary for maturation of tumor necrosis factor-alpha (TNF-α) converting enzyme, which is the molecule responsible for the release of TNF-α. In this study, Rhbdf2 knockout (KO) mice were produced by CRISPR/CAS9. To see the effects of the failure of TNF-α release induced by Rhbdf2 gene KO, collagen-induced arthritis (CIA), which is the representative TNF-α related disease, was induced in the Rhbdf2 mutant mouse using chicken collagen type II. The severity of the CIA was measured by traditional clinical scores and histopathological analysis of hind limb joints. A rota-rod test and grip strength test were employed to evaluate the severity of CIA based on losses of physical functions. The results indicated that Rhbdf2 mutant mice showed clear alleviation of the clinical severity of CIA as demonstrated by the significantly lower severity indexes. Moreover, a grip strength test was shown to be useful for the evaluation of physical functional losses by CIA. Overall, the results showed that the Rhbdf2 gene has a significant effect on the induction of CIA, which is related to TNF-α.
Animals ; Arthritis, Experimental* ; Chickens ; Collagen Type II ; Extremities ; Hand Strength ; Humans ; Joints ; Mice ; Mice, Knockout* ; Serine Proteases ; Tumor Necrosis Factor-alpha

A novel Helicobacter species was identified from the gastrointestinal tract of the Korean striped field mouse (Apodemus agrarius). Biochemical testing, ultrastructure characterization, and 16S rRNA gene sequence analysis suggested that this bacterium represents a distinct taxon. The bacterium was positive for urease activity, susceptible to cephalothin and nalidixic acid, and weakly positive for oxidase and catalase activity. Electron microscopy revealed that the bacterium has spirally curved rod morphology with singular bipolar nonsheathed flagella. Genotypically, the isolated bacterial strains (YMRC 000215, YMRC 000216, and YMRC 000419) were most closely related to a reference strain of Helicobacter mesocricetorum (97.25%, 97.32%, and 97.03% 16S rRNA sequence similarities, respectively). The 16S rRNA sequences of these strains were deposited into GenBank under accession numbers AF284754, AY009129, and AY009130, respectively. We propose the name Helicobacter apodemus for this novel species.
Animals ; Catalase ; Cephalothin ; Databases, Nucleic Acid ; Flagella ; Gastrointestinal Tract* ; Genes, rRNA ; Helicobacter* ; Korea* ; Mice* ; Microscopy, Electron ; Murinae* ; Nalidixic Acid ; Oxidoreductases ; Sequence Analysis ; Urease

The leptin receptor-deficient db/db mouse is a rodent model of type 2 diabetes and obesity. Diabetes in db/db mice shows an age-dependent progression, with early insulin resistance followed by an insulin secretory defect resulting in profound hyperglycemia. However, there is insufficient data on agedependent changes of energy metabolism in db/db mice. We demonstrated an age-dependent decrease in the respiratory exchange ratio (RER), calculated by a ratio of VO2/VCO2, in db/db mice. The RER determined by indirect calorimetry, was 1.03 in db/db mice under 6 weeks of age, which were similar to those in heterozygote (db/+) and wild-type (+/+) mice. However, RER decreased from approximately 0.9 to 0.8 by 10 weeks of age and subsequently returned to approximately 0.9 at 22 weeks of age. The changes in RER were concurrent with the alterations in body weight and blood glucose level. However, other metabolic indicators such as glucose tolerance, changes in body fat mass, and urinary glucose levels, did not change with age. The results suggested that the energy source utilized in db/db mice changed with the age-related progression of diabetes.
Adipose Tissue ; Animals ; Blood Glucose ; Body Weight ; Calorimetry, Indirect ; Energy Metabolism ; Glucose ; Heterozygote ; Hyperglycemia ; Insulin ; Insulin Resistance ; Leptin ; Mice* ; Obesity ; Rodentia

N-ethyl-N-nitrosourea (ENU) is a potent mutagen in a mouse model by inducing point mutation in a random manner and, in particular, causing heritable base substitutions in spermatogonia. In this study, systematic development of phenotype-driven mutant mice with large scale was carried out by using ENU. Nine-week-old male mice of C57BL/6J received intraperitoneal injection at three times with 100 mg/kg of ENU at weekly intervals for three weeks. After injections with ENU, the changes of body weight, fatality, recovery of fertile period, and breeding record were measured in these mice. Body weight lost as a result of ENU treatments was reversed after the last ENU injection. Live fertile male mice recovered from infertility from 104 to 165 days after ENU treatments were mated with C57BL/6J female mice for generation of G1 offspring. An average birth rate was 5.9 mice from 1 pair of paternal and maternal mice. All of 231 G1 offspring mice were analyzed by modified-SHIRPA with standard procedure at nine weeks of age. Among G1 mice, 166 mice were identified as mutagenic phenotypes in 20 test items. The changes in mutagenic phenotypes after ENU treatments, for instance, pattern in the region with a different color, touch escape, changes in head morphology, pupil, and teeth, and negative geotaxis etc., were found in these mice. Taken together, these results indicate that ENU may be a trans-generational mutagen in C57BL/6J mice.
Animals ; Birth Rate ; Body Weight ; Breeding* ; Ethylnitrosourea* ; Female ; Fertile Period* ; Head ; Humans ; Infertility ; Injections, Intraperitoneal ; Male ; Mice* ; Phenotype ; Point Mutation ; Pupil ; Spermatogonia ; Tooth ; United Nations

Vitamin D3 up-regulated protein 1 (VDUP1) is a potent growth suppressor that inhibits tumor cell proliferation and cell cycle progression when overexpressed. In a previous study, we showed that VDUP1 knockout (KO) mice exhibited accelerated liver regeneration because such animals could effectively control the expression of cell cycle regulators that drive the G1-to-S phase progression. In the present study, we further investigated the role played by VDUP1 in initial priming of liver regeneration. To accomplish this, VDUP1 KO and wild-type (WT) mice were subjected to 70% partial hepatectomy (PH) and sacrificed at different times after surgery. The hepatic levels of TNF-alpha and IL-6 increased after PH, but there were no significant differences between VDUP1 KO and WT mice. Nuclear factor-kappaB (NF-kappaB), c-Jun-N-terminal kinase (JNK), and signal transducer and activator of transcription 3 (STAT-3) were activated much earlier and to a greater extent in VDUP1 KO mice after PH. A single injection of TNF-alpha or IL-6 caused rapid activation of JNK and STAT-3 expression in both mice, but the responses were stronger and more sustained in VDUP1 KO mice. In conclusion, our findings provide evidence that VDUP1 plays a role in initiation of liver regeneration.
Animals ; Blotting, Western ; Carrier Proteins/*genetics/metabolism ; Cell Proliferation ; *Gene Expression Regulation ; Hepatectomy ; Hepatocytes/*cytology/physiology ; JNK Mitogen-Activated Protein Kinases/genetics/metabolism ; Liver/*physiology ; Male ; Mice, Knockout ; NF-kappa B/genetics/metabolism ; Polymerase Chain Reaction ; *Regeneration ; STAT3 Transcription Factor/genetics/metabolism ; Thioredoxins/*genetics/metabolism

The integrated cell culture-PCR (ICC-PCR) method has been suggested as an improved method for detection of viruses in water environments. We tested 57 source waters including finished water samples in Gyeonggi-do for enteric viral contamination using total culturable virus assay (TCVA) using BGMK cells and ICC-PCR. Nineteen of the 57 source water samples (33.3%) exhibited the cytopathic effect (CPE) on BGMK cells and no finished water did exhibited CPE. Nineteen samples (33.3%) of the 57 were positive for reoviruses. For the enteroviruses, only 3 samples (5.3%) of the 57 samples showed positive results. By using ICC-PCR method, 202 flasks from source water samples were positive for enteroviruses and reoviruses. Three samples from source water were positive for both viruses. However, any flasks tested was not co-infected with two types of viruses. While the enteric viral frequencies in TCVA and ICC-PCR were similar, the viral frequency for reoviruses at first passage in two type of method was higher in ICC-PCR (94.7%) than TCVA (56.9%).
Enterovirus ; Viruses ; Water

We investigated the effect of tilianin upon inducible nitric oxide synthesis in the plasma of low-density lipoprotein receptor knock-out (Ldlr-/-) mice fed with high cholesterol diet and in primary peritoneal macrophages of Ldlr-/- mice. High cholesterol diet induced nitric oxide production in the plasma of Ldlr-/- mice. Tilianin reduced the level of nitric oxide (NO) in plasma from Ldlr-/- mice induced by the high cholesterol diet. Tilianin also inhibited the NO production from the primary culture of peritoneal macrophages treated with lipopolysaccharide. The inhibition of NO production was caused by the suppression of inducible nitric oxide synthase (iNOS) gene expression in peritoneal macrophages isolated from Ldlr-/- mice. Moreover, tilianin inhibited the transcriptional activation of iNOS promoter that has NF-kappa B binding element. Thus, these results provide the first evidence that tilianin inhibit iNOS expression and production of NO and may act as a potential anti-inflammatory agent.
Tyrosine/analogs & derivatives/metabolism ; Tissue Distribution ; Sinus of Valsalva/metabolism/pathology/ultrastructure ; Receptors, LDL/*genetics ; Promoter Regions (Genetics)/drug effects ; Nitric Oxide Synthase Type II/*metabolism ; Nitric Oxide/biosynthesis/blood ; NF-kappa B/metabolism ; Mice, Knockout ; Mice ; Male ; Inflammation/metabolism ; Glycosides/*pharmacology ; Flavonoids/*pharmacology ; Down-Regulation/drug effects ; Atherosclerosis/metabolism ; Animals

BACKGROUND: Multiple myeloma is a clonal B-cell malignancy manifested by the accumulation of terminally differentiated plasma cells. The disease is characterized by clinical heterogeneity, with survival ranging from a few months to more than 10 years. The purpose of this study is to evaluate the prognostic value of specific chromosomal abnormality in multiple myeloma. METHODS: We analyzed the clinical records of 40 patients who were diagnosed as multiple myeloma, between April, 1995 and August, 2004. Cytogenetic analysis was conducted by metaphase karyotype analysis. Patients were grouped into normal cytogenetic group (arm A), complete or partial deletion of chromosome 13 and hypodiploidy group (arm B) and other cytogenetic abnormality group (arm C). RESULTS: Median follow up duration was 13.1 months (range 1.5-92.1). Overall response rate to chemotherapy was 58.8% and response rate among arm A, B and C were 56.3%, 33.3% and 75%, respectively (p=0.229). The prognostic factors affecting survival were clinical stage, performance status, serum creatinine level, sex and chromosomal abnormality. The median overall survival was significantly different among arm A, B and C (34.9 months, 8.5 months and 19.8 months, respectively, p=0.0125). CONCLUSION: chromosomal abnormality, especially, complete or partial deletion of chromosome 13 and hypodiploidy at initial diagnosis is significantly associated with survival duration.
Arm ; B-Lymphocytes ; Chromosome Aberrations* ; Chromosomes, Human, Pair 13 ; Creatinine ; Cytogenetic Analysis ; Cytogenetics ; Diagnosis ; Drug Therapy ; Follow-Up Studies ; Humans ; Karyotype ; Metaphase ; Multiple Myeloma* ; Plasma Cells ; Population Characteristics ; Prognosis

PURPOSE: This study was performed to identify risk factors for urinary tract infection(UTI) by extended-spectrum beta-lactamase(ESBL)-producing Escherichia coli in children. METHODS: We analyzed 125 episodes of UTI which were developed in 112 children at the Seoul National University Children's Hospital from July to December 2002. Medical records of 74 episodes of UTI caused by E. coli were reviewed in a search for possible risk factors of infection by ESBL producers. The presence of ESBL in E. coli was screened and confirmed by the National Committee for Clinical Laboratory Standards guidelines and/or double disk synergy test. RESULTS: The most common isolates were E. coli(74 isolates), followed by Enterobacter spp.(nine isolates) and Klebsillea spp.(eight isolates). Fifteen isolates were gram positive microorganisms. The prevalence of ESBL producers among 74 strains of E. coli was 18%(13 isolates). Previous hospitalization within six months(P=0.001; odds ratio[OR], 8.72, 95% confidence interval[CI], 1.87 to 40.6) and prior use of extended-spectrum cephalosporins within a month(P=0.009; OR,10.94, 95% CI, 1.59 to 75.4) were associated with the increased risk of UTI by ESBL-producing E. coli. However, age, sex, and the underlying urogenital anomalies did not contribute to the development of UTI by ESBL-producing E. coli. CONCLUSION: ESBL-producing strains of E. coli have been increasingly recognized in pediatric UTI. Previous use of antibiotics was related to the risk of infection by ESBL-producers. Therefore, prudent use of antibiotics should be recommended for antimicrobial therapy in children.
Anti-Bacterial Agents ; beta-Lactamases* ; Cephalosporins ; Child* ; Enterobacter ; Escherichia coli* ; Escherichia* ; Hospitalization ; Humans ; Medical Records ; Prevalence ; Risk Factors* ; Seoul ; Urinary Tract Infections* ; Urinary Tract*

OBJECTIVE: To evaluate the CT findings of ruptured corpus luteal cysts. MATERIALS AND METHODS: Six patients with a surgically proven ruptured corpus luteal cyst were included in this series. The prospective CT findings were retrospectively analyzed in terms of the size and shape of the cyst, the thickness and enhancement pattern of its wall, the attenuation of its contents, and peritoneal fluid. RESULTS: The mean diameter of the cysts was 2.8 (range, 1.5-4.8) cm; three were round and three were oval. The mean thickness of the cyst wall was 4.7 (range, 1-10) mm; in all six cases it showed strong enhancement, and in three was discontinuous. In five of six cases, the cystic contents showed high attenuation. Peritoneal fluid was present in all cases, and its attenuation was higher, especially around the uterus and adnexa, than that of urine present in the bladder. CONCLUSION: In a woman in whom CT reveals the presence of an ovarian cyst with an enhancing rim and highly attenuated contents, as well as highly attenuated peritoneal fluid, a ruptured corpus luteal cyst should be suspected. Other possible evidence of this is focal interruption of the cyst wall and the presence of peritoneal fluid around the adnexa.