No abstract available.
Adenoviridae* ; Child ; Child, Hospitalized* ; Coinfection* ; Humans ; Mycoplasma pneumoniae* ; Mycoplasma* ; Pneumonia* ; Pneumonia, Mycoplasma*

Human rotavirus has now been established as the leading cause of gastroenteritis in young children worldwide. At least fourteen serotypes of group A rotavirus have been identified on the basis of antibody responses to major neutralizing glycoprotein, VP7 (G type for glycoprotein), present in the outer capsid of the virus. Serotype 1, 2, 3 and 4 are the most highly prevalent in human. In Korea, rotavirus is also the principal cause of severe nonbacterial diarrhea requiring hospitalization in infants and young children, which is commonly detected by EIA method. The epidemiology of rotavirus infection has been monitored by only serologic methods without electropherotyping in Korea. This study shows seasonal and age related variations .of rotavirus infection in Korea according to the genotype using the reverse transcription polymerase chain reaction (RT-PCR). Fecal specimens were obtained from 39 children hospitalized with acute watery diarrhea and gastroenteritis in Ewha Womans University MokDong Hospital in Seoul from Jan. to Dec. of 1996. All four (1, 2, 3, 4) major G serotypes were identified by amplification of segment of the gene for VP7 using RT-PCR. Rotavirus Gl 749 bp, G2 653 bp, G3 374 bp and G4 583bp were shown on 2.9 or 3.3% NuSieve agar gel. Results were as follows: 1) Rotavirus was detected at 53.8% (21/39) by EIA and 89.7% (35/39) by RT-PCR. 2) Serotype Gl, G2, G3, G4 when detected by RT-PCR accounted for 80.0% (28/35), 14.3% (5/35), 2.9% (1/35) and 2.9% (1/35), respectively. 3) Thirty five strains of rotavirus were detected at the frequency of 17.1% (6/35) in Oct., 20.0% (7/35) in Nov. and 20.0% (7/35) in Dec. 4) As for the age range, children affected by rotavirus were mostly under 1 years.
Agar ; Antibody Formation ; Capsid ; Child ; Child, Hospitalized* ; Diarrhea* ; Epidemiology ; Female ; Gastroenteritis ; Genotype* ; Glycoproteins ; Hospitalization ; Humans* ; Infant ; Korea ; Polymerase Chain Reaction* ; Reverse Transcription* ; Rotavirus Infections ; Rotavirus* ; Seasons ; Seoul

Astrovirus is frequently associated with diarrhea in children. It can not be readily isolated by cell culture, and an electronmicroscope is usually used for detection of this agent. Recently in 1995 a combined method of reverse transcription-polymerase chain reaction (RT-PCR) was designed for easier detection of astrovirus, which is based on the conserved sequence in 3'-end of genomes of the 7 known serotypes of human astrovirus. As of yet there has not been any report of astrovirus data in Korea using the RT-PCR methods. The purpose of this study was to detect astrovirus incidence, severity of symptoms, seasonal variation and coinfection rate with rotavirus in Korean children inpatients with diarrhea. Fecal specimens from 61 young children hospitalized with gasteroenteritis Korea from Jan. 1996 through Mar. 1997. They were examined for astroviurs infection by RT-PCR method. Results are as follows: 1. Astrovirus was detected at 9.8% (6/61) from fecal specimens of children with severe diarrhea by EIA using monoclonal antibody coated plates. 2. Astorvirus was detected at 29.5% (18/61) from fecal specimens of children with severe diarrhea by RT-PCR. 3. The age of the 18 children affected by astrovirus ranged from 2 monthes to 7 years with mean of 3.0 years. 4. Mean hospital stay of the 1S children was 6.1 days. 5. Five (27.8%) astrovirus RT-PCR positive strains were confirmed in November and in December, respectively out of 18 specimens in total. 6. Astrovirus coinfection with rotavirus type G1 was confirmed in 15/16 specimens (93.8%), and with type G2 was in 1/16 specimens (6.3%).
Cell Culture Techniques ; Child* ; Coinfection ; Conserved Sequence ; Diarrhea ; Feces* ; Genome ; Humans ; Incidence ; Inpatients ; Korea ; Length of Stay ; Mamastrovirus ; Polymerase Chain Reaction* ; Reverse Transcription* ; Rotavirus ; Seasons

Apoptosis is a crucial mechanism for the selective elimination of mammalian cells and is involved in many physiological and pathological processes. The heightened awareness of the importance of apoptosis has increased the need for rapid and quantitative methods for measurement of apoptotic changes. Recently, 7-amino-actinomycin D (7-AAD) has been introduced as a valuable fluorescent dye for assessing apoptosis by flow cytometry. When the cells are stained with 7-AAD in the concentration of 10 - 20 ug/ml, live cells are not stained (7-AAD ) and early apoptotic cells are weakly stained (7-AAD ) while late apoptotic or dead cells are stained brightly (7-AAD). On scattergram of forward angle light scatter vs. 7-AAD fluorescence, the three populations can be discriminated not only between each other but also from cell debris or clumps. The apoptotic cells, defined as 7- AAD cells, were demonstrated as apoptotic by morphological observation of the sorted cells. The 7-AAD cell fraction was also demonstrated to be parallel with subdiploid fraction of cells stained with PL However, 7-AAD cells, whose definition is based on the alteration of membrane integrity, have never been demonstrated to be subdiploid fraction by simultaneous DNA staining. Here we directly demonstrate that 7-AAD cells, defined on the scattergram of forward angle light scatter vs. 7-AAD fluorescence, are subdiploid fraction by staining with DNA dye whose fluorescence is collected after 530/30 band pass filter (FL-1). We also demonstrate the effects of 7-AAD concentration, fixation of cells, and proliferation of cells, on the fluorescence pattern, for reference during assessment of apoptosis by simple and rapid method for flow cytometric analysis of cells stained with 7- AAD. We also present a flow cytometric analysis of cells stained with 7-AAD Eor sequential change in apoptotic fraction, with concurrent dual-color immunophenotyping.
Apoptosis* ; DNA ; Flow Cytometry* ; Fluorescence ; Fluorescent Antibody Technique* ; Immunophenotyping ; Membranes ; Pathologic Processes

PURPOSE: HL-60 is a promyelocytic cell line. Fc receptors and complement receptor 3 (CR3) play important role in the protective response of granulocytes and monocytes against microbial infection. We quantified the expression of Fc I, Fc II, Fc III, and CD11b/CD18 during differentiation using HL-60 cells by N-N-dimethylformamide (DMF). Functional studies, such as phagocytic activity, respiratory burst and ADCC, were also performed. METHODS: HL-60 cells were induced to differentiate by adding 0.8% DMF. On the 4th and 7th day after stimulation as well as before stimulation, the cells were analyzed for phenotypic and functional differentiaton. Phenotypic analysis was performed by flow cytometry after staining the cells with PE-conjugated anti-human CD64, CD32, CD16, CD11b, CD18, and isotype controls. And the measured fluorescent intensity was transformed into Molecules of Equivalent Soluble Fluorochromes (MESF). Phagocytic activity was also measured by flow cytometry after incubation with fluorochrome-conjugated beads. Respiratory burst was measured by chemiluminescence assay of cells incubated with luminol after stimulation with PMA. ADCC was measured by hemoglobin release assay. RESULTS: The expression of CD11b, CD18 and CD64 on HL-60 cells markedly increased on the 4th day and slightly decreased on the 7th day. Expression of CD32 was already induced before differentiation induction and slightly increased by DMF. CD16 was not expressed during differentiation. In phagocytic assay, the phagocytic cell fraction increased by stimulation on 4th and 7th day. Chemiluminescence showed the DMF increased the respiratory burst of HL-60 cells on the 4th and 7th day. In ADCC, DMF increased the target cell lysis continuously. CONCLUSION: HL-60 cells which were differentiated with DMF for are good models for studying opsonophagocytic assay of immunized sera.
Antibody-Dependent Cell Cytotoxicity ; Cell Line* ; Flow Cytometry ; Fluorescent Dyes ; Granulocytes ; HL-60 Cells ; Humans ; Luminescence ; Luminol ; Monocytes ; Phagocytes ; Receptors, Complement ; Receptors, Fc ; Respiratory Burst

Varicella-Zoster virus (VZV) is the causative agent of chickenpox in children. It also causes herpes zoster in old or immunocompromised people. For effective control of VZV, live attenuated vaccine (Oka strain) was developed in 1974 and has been used worldwidely. This vaccine is indicated when VZV infection can be fatal, such as leukemia, with favorable success. Recently, routine usage of VZV vaccine for general young population was proposed. However, general use of vaccine requires prior epidemiological study of wild strains prevailing in the target population. In this regard, we investigated the antigenic and genetic variations, if any, between Oka strain and wild strains isolated in Korea. Six wild strains of VZV isolated from zoster patients in Korea, Ellen (the labaratory-adapted strain) and Oka (vaccine) strains were cultivated in Vero cells. After the VZV-infected Vero cells were stained with specific monoclonal antibody panel, the immunofluorescent patterns were compared. The VZV-infected Vero cells were also extracted after metabolic labelling to be immunoprecipitated with the monoclonal antibodies. However, the patterns of immunofluorescent staining and immunoprecipitation did not show any antigenic difference among wild strains and vaccine or standard strain. When the VZV gene was amplified by polymerase chain reaction using primer VZV 115953 and 116605, and the PCR products were cleaved by restriction enzymes (Taq I, Bl II, and Hpa II), two kinds of restriction patterns were observed. These results suggested that antigenic variation was not common among wild strains of VZV isolated in Korea and between vaccine strain. But the different restriction patterns implied potential antigenic variation.
Antibodies, Monoclonal ; Antigenic Variation ; Chickenpox ; Child ; Epidemiologic Studies ; Genetic Variation ; Health Services Needs and Demand ; Herpes Zoster ; Herpesvirus 3, Human* ; Humans ; Immunoprecipitation ; Korea* ; Leukemia ; Polymerase Chain Reaction* ; Polymorphism, Restriction Fragment Length* ; Vero Cells

No abstract available.
Colorectal Neoplasms* ; Epidermal Growth Factor* ; Humans ; Receptor, Epidermal Growth Factor*

No abstract available.
Epithelial Cells* ; Salmonella typhimurium* ; Salmonella*

No abstract available.
Antibodies, Monoclonal* ; Glycoproteins* ; Humans*

For the purpose of morphologic analysis of human cytomegalovirus (HCMV) antigens reacting with specific monoclonal antibodies, we observed HCMV particles after immunogold staining. HCMV was cultured in human fetal lung fibroblasts to be concentrated by polyethylene glycol 6,000. The HCMV stock was dropped onto Formva-coated grids and was fixed by 2% glutaraldehyde. The grids were reacted with MCMVA57, 93, 135 or with SCMVM1, 6, 14, 49 monoclonal antibodies (MoAbs) follwed by gold (10 nm)-conjugated goat anti-mouse IgG. Then the grids were stained with 2.5% uranyl acetate to be observed under Hitachi 500 or Jeol 1,200 electron microscope. When HCMV was reacted with SCMVM14 and SCMVM49 MoAbs, gold particles were adsorbed to virion envelopes, suggesting that the reactive antigens were envelope proteins. In cases of MCMVA135 and SCMVM6 MoAbs, gold particles were adsorbed to dense bodies as well as to virion envelope. These results, together with the previous results of immunologic and genetic characterization, suggested that the reactive antigens of MCMVA135 and SCMVM6 MoAbs were gB homologue and structural protein, respectively. In case of SCMVM1 MoAb, gold particles were adsorbed to capsids, envelopes, and dense bodies, suggesting that the reactive antigen was structural protein. In case of SCMVM8 MoAb, gold particles were observed between the envelopes and capsids, which space was supposed to be the tegument, suggesting that the reactive antigen was carbohydrate moiety of glycoprotein or its polymer. In cases of MCMVA57 and MCMVA93 MoAbs, gold particles were adsorbed to only dense bodies, suggesting that the reactive antigens were precursors of structural proteins.
Antibodies, Monoclonal* ; Capsid ; Cytomegalovirus* ; Fibroblasts ; Glutaral ; Glycoproteins ; Goats ; Humans* ; Immunoglobulin G ; Lung ; Polyethylene Glycols ; Polymers ; Virion