Experimental autoimmune encephalomyelitis was induced in macaques. T cell clones infiltrated into the brain lesion area were compared with those in blood. Intradermal immunization of macaques with brain white matter derived from healthy macaque in combination with pertussis toxin, induced neurological symptoms in two macaques. One died on day 25 after immunization, whereas the other survived. Gross examination of the brain from the dead macaque, showed clear hemorrhagic lesions in the white matter. Hematological analysis showed that drastic T cell response was induced in macaques immunized with white matter, but not in control macaques. Flow cytometric analysis of blood cells from the affected macaques demonstrated an increase of CD4 and CD8 T cell populations expressing the CD69 early activation marker. Single strand conformation polymorphism (SSCP) analysis of T cell receptor beta chain showed T cell clones infiltrated into the brain lesion, which were different from those found in the peripheral blood of the same monkey. The present paper shows that SSCP analysis of TCR is useful in studying clonality of T cells infiltrating into the brain tissue of macaque with EAE.
Animals ; Antigens, CD3/analysis ; Disease Models, Animal ; Encephalomyelitis, Autoimmune, Experimental/immunology/*pathology ; Flow Cytometry/veterinary ; Leukocyte Count/veterinary ; *Macaca fascicularis ; Male ; Polymorphism, Single-Stranded Conformational ; Receptors, Antigen, T-Cell, alpha-beta/genetics ; T-Lymphocytes/cytology/immunology

In Drosophila, rhomboid proteases are active cardinal regulators of epidermal growth factor receptor (EGFR) signaling pathway. iRhom1 and iRhom2, which are inactive homologs of rhomboid intramembrane serine proteases, are lacking essential catalytic residues. These are necessary for maturation and trafficking of tumor necrosis factor-alpha (TNF-α) converting enzyme (TACE) from endoplasmic reticulum (ER) to plasma membrane through Golgi, and associated with the fates of various ligands for EGFR. Recent studies have clarified that the activation or downregulation of EGFR signaling pathways by alteration of iRhoms are connected to several human diseases including tylosis with esophageal cancer (TOC) which is the autosomal dominant syndrom, breast cancer, and Alzheimer's disease. Thus, this review focuses on our understanding of iRhoms and the involved mechanisms in the cellular processes.
Alzheimer Disease ; Breast Neoplasms ; Cell Membrane ; Down-Regulation ; Drosophila ; Endoplasmic Reticulum ; Esophageal Neoplasms ; Humans ; Keratoderma, Palmoplantar, Diffuse ; Ligands ; Peptide Hydrolases ; Receptor, Epidermal Growth Factor ; Serine Proteases ; Tumor Necrosis Factor-alpha

No abstract available.
Epidemiology* ; Measles*

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

PURPOSE: Invasive bacterial infection is a major cause of morbidity and mortality in children. Previously, we reported etiology of invasive infections in healthy children in 1985-1995. This study was performed to update etiology of invasive bacterial infections in the previously healthy children. METHODS: We reviewed medical records of 98 episodes of invasive bacterial infections in immunocompetent children at the Seoul National University Children's Hospital in 1996-2004. RESULTS: The frequent pathogens identified over all age groups were Streptococcus pneumoniae (33 %) and Staphylococcus aureus (33%). The proportion of Salmonella species and Haemophilus influenzae has been declined to 4% each from 23% and 14%, respectively, compared to previous study. S. agalactiae was the most common isolate in the infants < or =3 months. Among the infants and children aged 3 months to 2 years and children of 2-5 years, S. pneumoniae (57%, 52%, respectively, in each group) was the most common isolates followed by S. aureus (17% and 24%, respectively). S. aureus was the most common isolates (73%) in children > 5 years. Primary bacteremia was the most common clinical diagnosis (27%). S. pneumoniae was responsible for 42% of primary bacteremia, 50% of meningitis, and 69% of bacteremic pneumonia and empyema. S. aureus accounted for 80% of bone and joint infections. The case fatality rate was 8.1% for all invasive infections. CONCLUSION: We reviewed frequency of bacterial agents of invasive infections in children. The data may be useful for pediatricians to select adequate empirical antibiotics in the management of invasive bacterial infections.
Infant ; Child ; Male ; Female ; Humans ; Mortality

Embryonic stem cells (ESCs) are an emerging source for cell-based therapies aimed at repairing damaged organ tissues; however, the efficiency of directed differentiation is low and refinement of differentiation protocols is hampered by incomplete understanding of the mechanisms involved in this process. To find new compounds which can improve the efficiency of directed differentiation of ESCs to cardiomyocytes, we screened several thousand chemical compounds and identified a promising group. All of the compounds found have a common structure of 1H-pyrrole,2,2'-(phenylmethylene)bis. Here we report the potential mechanism of action for 31002 which showed the strongest activity among the compounds selected. In the presence of 31002, 15 times more cardiomyocytes differentiated from ESCs, i.e., 3.5% to 52% of total differentiated cells. Moreover, the cardiomyocytes showed functional characteristics including rhythmic beating and marker gene expression. 31002 inhibited the down-regulation of genes related to the three germ layers in the late stage of ESCs differentiation, implying that 31002 supports a continuous fate commitment of undifferentiated ESCs to the cardiac lineage by prolonging the three germ layer stages. Therefore, compounds in this group, including 31002, might be useful as directed cardiomyogenic differentiation-inducers to produce cells for use in cell therapy aimed at restoring damaged heart tissue.
Down-Regulation ; Embryonic Stem Cells ; Gene Expression ; Germ Layers ; Heart ; Myocytes, Cardiac ; Tissue Therapy

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

PURPOSE: Streptococcus pneumoniae is a major etiologic agent for pneumonia, meningitis, otitis media, and sepsis among young children. Multi-drug resistant strains have raised great concern worldwide, thus the importance of prevention with vaccines has been emphasized. However, vaccines may force the appearance of pneumococcal infections by nonvaccine serotypes. Thus, distribution of pneumococcal serotypes should be monitored to estimate vaccine efficacy. We used a new and efficient multibead assay in determining pnemococcal serotypes. METHODS: From January to February 2005, 643 children were recruited from ten day care centers to isolate pneumococci from their oropharynx. Pneumococcal serotyping was performed on 62 pneumococcal isolates from 60 children by multibead assay. This immunoassay required two sets of latex particles coated with pneumococcal polysaccharides and serotype-specific antibodies. Twenty four newly developed monoclonal antibodies specific for common serotypes and a pool of polyclonal rabbit sera for some of the less common serotypes were used. RESULTS: The most prevalent pneumococcal serotypes were serotype 6A, 19A, 19F, 23F, and 11A/ D/F which accounted more than 50 precent of all the 62 pneumococcal isolates. We found that multibead assay can be performed very rapidly and objectively. CONCLUSION: This multibead immunoassay was very useful in serotyping clinical isolates of S. pneumoniae because it was simple, reliable and fast.
Antibodies ; Antibodies, Monoclonal ; Child ; Day Care, Medical ; Humans ; Immunoassay ; Meningitis ; Microspheres ; Oropharynx ; Otitis Media ; Pneumococcal Infections ; Pneumonia ; Polysaccharides ; Sepsis ; Serotyping* ; Streptococcus pneumoniae* ; Streptococcus* ; Vaccines

PURPOSE: Pneumoccocus is one of the most important causes of invasive infection through the childhood period and the prevelance of antibiotics resistance of pneumococcus is increasing worldwide. A 7-valent conjugate vaccine has been developed. It is important to know the prevalence of each serotype of pneumococci in the countries where the vaccine is used to estimate the coverage rate by the vaccine. METHODS: One hundred and twenty seven strains of clinical isolates and 72 strains from healthy carriers recovered from Korean children during the period from 1997 to 2002 were subjected to determination of serotype by Quellung reaction and penicillin susceptibility with oxacillin disc diffusion test. RESULTS: Forty-three per cent of clinical isolates were obtained from children under two years of age. Thirty strains(24%) were isolated from normally sterile body fluids. The frequent serotypes were 19F, 19A, 23F, 6A, 6B and 9V. Fifty-six per cent of the clinical isolates were represented in the current 7-valent protein conjugate pneumococccal vaccine, and 84% when the cross-reactive serotypes were included. Frequent serotypes of strains isolated from one to five year-old healthy children were 19F, 14, 11A, 23F, 18C, and 19A. Seventy-one per cent of the carrier strains were included in the 7-valent vaccine. Ninety-three per cent of the clinical isolates and 86% of carrier strains were not susceptible to penicilline. CONCLUSION: Fifty-six to 84% of pneumococci recovered from Korean children are covered by the current 7-valent protein conjugate pneumococcal vaccine and the prevalence of penicillin resistance was very high.
Anti-Bacterial Agents ; Body Fluids ; Child* ; Diffusion ; Humans ; Oxacillin ; Penicillin Resistance ; Penicillins* ; Prevalence ; Streptococcus pneumoniae* ; Streptococcus*