Purpose of the present study was to find the optimal ovulation induction medicine for the maturation and development of immature oocytes and culture media for 2-cell embryos in the mouse model. ICR female mouse aged 6 to 8 weeks, were stimulated with 5 IU PMSG injection. At 47 to 50 hour post-PMSG injection, ovaries were dissected out and oocytes-cumulus complexes were punctured. The oocyte-cumulus complexes were cultured in media containing various ovulation induction medicine, CC, HMG and Metrodin for 18 hours. Female ICR mice were stimulated with 5 IU PMSG and 48 hours later were injected 5 IU of hCG, then female and male mice were mated. At 48 hour post-hCG injection, oviducts were dissected out and 2-cell embryos were flushed. The 2-cell embryos were cultured in various media, Ham's F-10 media of milli-Q water (3degrees), Ham's F-10 media of HPLC (high performance liquid chromatography, Baxter) water, Medicult media, HTF (human tubal fluid) media for 96hours. The results were as follows. 1. When the oocytes-cumulus complexes were cultured in 10(-9)microgram/ml~ 10(-8)microgram/ml of CC, those were suppressed in meiotic maturation (28.2~ 33.7%). Whereas the oocytes-cumulus complexes were cultured in 10(-7)microgram/ml~10(-4)microgram/ml, these were not effected in meiotic maturation (54.5~72.7%). 2. When the oocytes-cumulus complexes were cultured in 10(-4)microgram/ml~ 10(-1)microgram/ml of Metrodin, those were suppressed in meiotic maturation (35.7~ 41.5%). Meanwhile the oocytes-cumulus complexes were cultured in 10(-7)microgram/ml~10(-5)microgram/ml, those were not effected in meiotic maturation (54.2~ 70.3%). 3. When the oocytes-cumulus complexes were cultured in 10(-5)microgram/ml~ 10(-4)microgram/ml of HMG, those were suppressed in meiotic maturation (48.2~ 50.4%). As being cultured in 10(-7)microgram/ml~10(-6)microgram/ml, increased in meiotic maturation (75.8~80.7%). 4. When the 2-cell embryos were cultured in Ham's F-10 media of milli-Q wats. ( 3degrees), Ham's F-10 media of HPLC (high performance liquid chromatograpy, Banter) water, Medicult media, HTF (human tubal fluid) media, developmental rates to blastocyst and hatching for 96 hour were 50.0%, 45.2%, 71.5% and 95.6%, respectively.
Animals ; Blastocyst ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Culture Media ; Embryonic Structures* ; Female ; Humans ; Male ; Mice* ; Mice, Inbred ICR ; Oocytes* ; Ovary ; Oviducts ; Ovulation Induction* ; Ovulation* ; Urofollitropin ; Water

Cryopreservation is able to store the surplus pre-embryos for freezing and furthermore thawing and transfer in a subsequent cycle. Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps, freezing media and embryonic stages on the rates of viability and development of cryopreserved mouse embryos. Female ICR mice (6~8 weeks old) were induced to superovulate by sequential intraperitoneal injection of 5 IU PMSG and 5 IU hGC 48h apart. Mouse embryos were collected according to its developmental stage after the injection of hCG. Embryos were cryopreserved not only by cryoprotectant step (1 step~ 4 step) but also in a variety of media (HTF, IVF medium, D-PBS) and cell stage. The results were as follows: There is no clear advantage in these freezing media of rapid method, but 4 cell and 8 cell of slow method (2, 3, 4 step) have advantage in D-PBS. The development of embryos according to cell stage become greater in 8 cell stage. In the treatment steps of cryopreservation, the development of embryo to blastocyst was similar among rapid method, but the development of 4 cell and 8 cell embryos to blastocyst according to slow method was better than rapid method.
Animals ; Blastocyst ; Cryopreservation* ; Culture Media* ; Embryonic Structures* ; Female ; Freezing ; Humans ; Injections, Intraperitoneal ; Mice* ; Mice, Inbred ICR

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

Huntington's disease is caused by CAG trinucleotide expansions in the gene encoding huntingtin. N- terminal fragments of huntingtin with polyglutamine produce aggregates in the endosome-lysosomal system, where proteolytic fragments of huntingtin is generated. Heat shock protein 70 (HSP70) prevents the formation of protein aggregates, but the effect of HSP70 on the huntingtin in the endosome-lysosomal system is unknown. This study was to determine whether HSP70 alters the distribution of huntingtin in endosome-lysosomal system. HSP70 expressing stable cells (NIH/3T3 or cerebral hybrid cell line A1) were generated, and mutant [(CAG)100] huntingtin was transiently overexpressed. Analysis of subcellular distribution by immnuocytochemistry or proteolysis cleavage by Western blotting was performed. 18 CAG repeat wild type [WT; (CAG)18] huntingtin was used as a control. Cells with huntingtin showed patterns of endosome- lysosomal accumulation, from a 'dispersed vacuole (DV)' type into a coalescent 'perinuclear vacuole (PV)' type over time. In WT huntingtin, HSP70 increased the cells with the PV types that enhanced the proteolytic cleavage of huntingtin. However, HSP70 reduced cells of the DV and PV types expressing mutant huntingtin, that result in less proteolysis than that of control. In addition, intranuclear inclusions were formed only in mutant cells, which was not affected by HSP70. These results suggest that HSP70 alters the distribution of huntingtin in the endosome-lysosomal system, and that this contributes to huntingtin proteolysis.
Peptide Hydrolases/metabolism ; Nuclear Proteins/genetics/*metabolism ; Nerve Tissue Proteins/genetics/*metabolism ; NIH 3T3 Cells ; Mice ; Lysosomes/*metabolism ; HSP70 Heat-Shock Proteins/genetics/*metabolism ; Endosomes/*metabolism ; Cytoplasm/metabolism ; Cell Survival ; Animals