OBJECTIVES: This study was to investigate the fertilization rate after intracytoplasmic sperm injection (ICSI) or partial zona dissection (PZD) of human and hamster sperm into hamster oocyte in in vitro fertilization (IVF). In addition, the possibility of clinical application was evaluated by the comparison of usefulness and difference of these method. MATERIALS AND METHODS: Hamster immature oocytes were obtained from oviducts superovulated by PMSG and hCG, and hamster sperms were obtained from epididymis. The freezed human sperms were thawed before use. Fertilization were confirmed by two pronuclei, one pronucleus, swollen sperm head or/and two polar bodies at 7~8 hour after ICSI or PZD. RESULTS: The fertilization rates after ICSI and PZD of human sperm to hamster oocyte were 3.6%, 64.2%, 73.6%, and 55.6% for negative control, positive control, ICSI, and PZD respectively, suggesting that ICSI only showed improved fertilization rate (p<0.01). The fertilization rates after ICSI and PZD of hamster sperm to hamster oocyte were 11.1%, 51.2%, 39.6%, and 72.7% for negative control, positive control, ICSI, and PZD respectively, suggesting that PZD only showed improved fertilization rate (p<0.01). PZD showed significantly higher fertilization rate than ICSI (p<0.05). CONCLUSIONS: As for the fertilization rate by ICSI and PZD using hamster oocyte in IVF, ICSI technique was considered to be more useful for human sperm and PZD technique for hamster sperm. Therefore, ICSI technique was considered more appropriate for experimental application using human sperm and hamster oocyte.
Humans

OBJECTIVES: This study was carried out to evaluate the effects of embryonic stage, cryoprotectant, and freezing-thawing method on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). MATERIALS AND METHODS: Two to eight cell embryos were obtained from oviducts of mated F1 hybrid female mice superovulated by pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Two-step 1,2-propanediol (PROH), dimethylsulfoxide (DMSO) and 4-step PROH, DMSO were used as cryoprotectant and dehydration and rehydration method of embryos, and slow- cooling or rapid-cooling method was used as frozen program. The survival rates of embryos were measured after thawing and rehydration, and the developmental rates of embryos were compared and observed during culturing embryos for 24, 48, 72, 96 hrs. RESULTS: As for the survival and development rates of embryos according to embryonic stage, the survival rate of 2 cell stage in PROH and DMSO was significantly higher than 4-8 cell (64.5% versus 62.1%, 79.7% versus 73.2%) (p<0.01, p<0.01), but the development rates of 4-8 cell embryos in PROH and DMSO were significantly higher than 2 cell embryos for whole culture period (p<0.01) and the development rates of 4-8 cell embryos in PROH were significantly higher than 2 cell embryos in DMSO (p<0.01). As for the survival and development rates of embryos according to cryoprotectant, the survival rate of 2 cell embryo in DMSO was significantly higher than that in PROH (74.4% versus 64.5%) (p<0.01), whereas the development rate of embryos was not differ till 24 hrs. The development rate from morular to hatching blastocyst, however, was significantly higher in PROH than in DMSO during 48 hr (p<0.01). The survival rate of 4-8 cell embryo was 62.1% in PROH and 73.2% in DMSO. The development rates of embryo in PROH were significantly higher for whole culture periods (p<0.01, 0.05). In respect to the effect of freezing and thawing program on the survival and development rates of embryos, method of slow cooling and rapid thawing was more effective than that of rapid cooling and rapid thawing. CONCLUSIONS: The survival rate of embryo in 2 cell stage was higher than in 4-8 cell stage, and PROH appears more effective cryoprotectant than DMSO because PROH showed better development rates of embryos in 2 and 4-8 cell stage. Moreover, slow cooling and rapid thawing method was considered as the best cryopreservation program.
Pregnancy ; Female ; Humans ; Mice ; Animals

No abstract available.
Peroxisomes

Epidermoid cysts occur rarely on the palms and soles, where hair is not present. These cysts have long been assumed to arise from traumatic implantation of epidermal fragment, but the pathogenesis is still controversial. Recently, with microscopic findings, immunohistochemical features, and molecular studies, the epidermoid cysts of the sole may be induced by the human papillomavirus. We report two cases of epidermoid cysts of the sole with the discussion of the pathogenesis.
Epidermal Cyst* ; Hair ; Humans*

The universal first step for the treatment of diabetes mellitus, hypertension, dyslipidemia or atherosclerotic disease is therapeutic lifestyle change (TLC). However, in most cases, long-term sincere maintenance of TLC is very difficult. The Look-AHEAD study was initiated based on the results of the Diabetes Prevention Program (DPP) trial that evaluated the effects of TLC on the prevention of diabetes. However, the look-AHEAD trial failed to demonstrate the benefit of intensive lifestyle intervention (ILI) for the prevention of CVD or CVD-associated mortality. Several explanations were suggested, including that the ILI group did not maintain effective weight loss, and that the intervention and control groups both received the same high-quality medical treatments. The participants of the look-AHEAD trial were very highly motivated, and the investigators of the trial systematically pursued their patients to achieve the desired end result, which is quite different from the usual clinical practice setting. Simple advisement for patients to perform diet restriction and moderate intensity regular physical exercise typically fails and is ineffective for the prevention of CVD in actual clinical practice. Concomitant sincere control of other risk factors is necessary for long-term CVD prevention. Further research on practical and effective methods for achievement of successful TLC is needed.
Cardiovascular Diseases* ; Diabetes Mellitus ; Diet ; Dyslipidemias ; Exercise ; Humans ; Hypertension ; Life Style* ; Mortality* ; Research Personnel ; Risk Factors ; Weight Loss

Objectives: This study was to evaluate whether Ham's F-10 used in assisted reproductive technology (ART) could be replaced with newly-introduced Medicult or Human Tubal Fluid (HTF) media, and the rate of embryo development could be enhanced by cumulus cell coculture. METHODS: Ham's F-10, Medicult, and HTF media supplemented with 0.4% bovine serum albumin (BSA) were used. Two-cell embryos were obtained from oviducts of mated F1 hybrid female mice superovulated by pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Cumulus cells for coculture were obtained from oviducts of ICR female mice superovulated by PMSG and hCG. Two-cell embryos were cultured in Ham's F-10, Medicult, and HTF media respectively to observe and compare the rate of embryo development. In addition, two-cell embryos were cultured in these three media for 24, 48, 72, 96 hrs with or without cumulus cell, and rates of embryo development were investigated and compared. RESULTS: As for the rate of embryo development to hatched blastocyst after 96 hrs culture, HTF (87.5%) and Ham's F-10 (85%) were significantly higher than Medicult (70.5%). The beneficial effect of embryo development by cumulus cell coculture on two-cell mouse embryo among these three media was enhanced significantly in Medicult (control 88.5% versus coculture 98.5%) by 24 hrs, and was not enhanced statistical significantly but slightly elevated in Ham's F-10 (86.5% versus 95.5%) and HTF (91.3% versus 96.9%) by 48 hrs, but rates of embryo development were similar between control and coculture group in all three media by 96 hrs. Significant differences were not shown in three media, but HTF showed generally high tendency of the enhancing effect of embryo development and the beneficial effect of embryo development by coculture. CONCLUSIONS: As a result of culturing two-cell embryos in three media for 96 hrs, generally HTF and Ham's F-10 showed higher rate of embryo development than Medicult. As for the beneficial effect of coculture, Medicult only showed early transient significant improvement of embryo development. Considering that coculture effect of good quality media may be not so great, Ham's F-10 and HTF are more stable media than Medicult. Accordingly, HTF may be considered to be a medium to replace with Ham's F-10, however, the present study suggest that Medicult or HTF is not able to replace with Ham's F-10 in ART.
Animals ; Blastocyst ; Chorionic Gonadotropin ; Coculture Techniques* ; Culture Media* ; Cumulus Cells* ; Embryonic Development* ; Embryonic Structures* ; Female ; Gonadotropins ; Humans* ; Mice* ; Oviducts ; Pregnancy ; Reproductive Techniques, Assisted ; Serum Albumin, Bovine

No abstract available.
Factor V Deficiency* ; Factor V*

OBJECTIVE: Immunofluorescence microscopy including confocal laser scanning microscopy and electron microscopy were used to study the production of fibronectin, tenascin, and laminin in the cumulus-corona (CC) cells surrounding mature, unfertilized oocytes after ovulation in view of their presumptive importance in the coordination of the processes leading to fertilization and early embryo cleavage, including the final maturation of the ovum, the sperm-egg interaction, and the complex biochemical mechanism between the ovum and the oviduct. METHODS: Mature oocyte-cumulus complex (OCC) was cultured for 24 and 48 hour and fixed in 3.7% formaldehyde. Specimens were incubated with a mixture of primary monoclonal antibodies recognizing different epitopes of fibronectin, tenascin, and laminin, and then with a mixture of secondary antibodies containing FITC, TRITC, and Cy-5 conjugated antibodies. Observation was made by confocal laser scanning microscope equipped with epifluorescece optics. Transmission electron microscopy were used to observe the OCC at 24 and 48 hours after cultrue. RESULTS: The immunocytochemical date demonstrated that CC masses are capable of producing fibronectin and tenascin but their production is heterogeneous in the CC population. Immunoreactivity to fibronectin and tenascin was shown mostly by inner corona cells, and the intensity of immunofluorescence decreased from the central corona cells to the peripheral cumulus cells. Colocalization of fibronectin and tenascin was evident in most CC cells. Moreover, fibronectin and tenascin immunoreactive material was observed in the intracytoplasmic areas, at the plasma membrane level as well as in the extracellular matrix. Whereas, laminin immunofluorescence was found around plasma membrane and extracellular area, but a intracytoplasmic reaction was rarely observed. The distribution of laminin immunofluorescence was similar to that of fibronectin and tenascin, but in some cumulus cells, colocalization between them was not found. Ultrastructurally, cumulus cells projected numerous long, thin microvilli into the intercellular area and some micovilli penetrated into zona pellucida. The inner layer of the cumulus mass was loose arrangement of relatively uniform, small cells with widened intercellular spaces, whereas in the outer layer, cumulus cells are rather larger in size and compact arrangement by narrow, irregular spaces. A small and large linear gap junctions were easily found at cell contacts. The cytoplasm of most cells had abundant organelles typical of steroidogenesis: numerous mitochondrias, a well-developed smooth endoplasmic reticulum, electron dense lipid droplets, and bundles of microtubules and microfilaments. Rudimentary disrupted basal lamina along the cytoplasmic border was rarely seen in a few inner conora cells. CONCLUSION: Even though the functional role of these extracellular matrix proteins remains still unclear, it is reasonable to suggest that they are necessary in various steps of the reproductive process. Cumulus cells appears to be a heterogeneous and dynamic system for suitable microenviroment of fertilization. And functional differences between corona and cumulus cells during the oocyte denudation may be accounted for particular distribution of these adhesive proteins and steroidogenesis-related organelles.
Actin Cytoskeleton ; Adhesives ; Animals ; Antibodies ; Antibodies, Monoclonal ; Basement Membrane ; Cell Membrane ; Cumulus Cells ; Cytoplasm ; Embryonic Structures ; Endoplasmic Reticulum, Smooth ; Epitopes ; Extracellular Matrix ; Extracellular Matrix Proteins ; Extracellular Space ; Female ; Fertilization ; Fibronectins* ; Fluorescein-5-isothiocyanate ; Fluorescent Antibody Technique ; Formaldehyde ; Gap Junctions ; Immunohistochemistry ; Laminin* ; Microscopy, Confocal ; Microscopy, Electron* ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; Microtubules ; Microvilli ; Mitochondria ; Oocytes ; Organelles ; Oviducts ; Ovulation ; Ovum ; Sperm-Ovum Interactions ; Tenascin* ; Zona Pellucida

No abstract available.
Anemia, Hypoplastic, Congenital*

No abstract available.