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Biomedical subjects

Y Fukui

Publications and source records attributed to Y Fukui.

At least 469 records · Page 26Linked to original sources
PubMed1983

Fertilization and cleavage of bovine follicular oocytes in rabbit reproductive tracts after maturation in vitro.

Bovine follicular oocytes cultured in vitro in Ham's F-12 based medium for 27 hours were transferred to the oviducts of rabbits pretreated with 75 IU hCG 24 hours previously. Frozen-thawed bull spermatozoa were inseminated into the uterus of rabbits 0, 2, 4, 6, 9, and 12 hours before oocyte transfer. Fertilization and cleavage were examined in the oocytes recovered from the oviduct and uterus 24 and 48 hours later. The criteria of fertilization were the presence of an enlarged sperm head, sperm tail, and pronuclei in the vitellus. Bull spermatozoa inseminated 4 hours before oocyte transfer showed the highest rates of fertilization at both 24 and 48 hours (35% and 38%) compared to the other preincubation times. The numbers of cleaved oocytes were also greater at these times. The results indicate that the capacitation time of cryopreserved bull spermatozoa in the rabbit reproductive tract is the same as in the bovine tract--i.e., 4-6 hours.

Animals↗
PubMed1983

The 19-kDal protein encoded by early region 1b of adenovirus type 12 is synthesized efficiently in Escherichia coli only as a fused protein.

We have constructed recombinant plasmids that direct the synthesis of the Mr 19 000 protein encoded by the adenovirus type 12 (Ad12) E1b region as either a native protein or a protein fused to the amino-terminal portion of the elongation factor EF-TuB in Escherichia coli cells. Using these recombinants, we could synthesize a large amount of the fused protein, while only a small amount of the native Mr 19 000 protein was produced. The failure to synthesize the native Mr 19 000 protein in E. coli cells was ascribed to inefficient translation.

Adenoviridae↗
PubMed1983

Filopodelike projections induced with dimethyl sulfoxide and their relevance to cellular polarity in Dictyostelium.

When 5% dimethyl sulfoxide (DMSO) was applied to Dictyostelium cells, the cells rounded up in shape and cytoplasmic streaming ceased. The cells resumed both cytoplasmic streaming and locomotion in 20 min. SDS PAGE of isolated plasma membrane fractions showed that actin and myosin apparently became dissociated from the plasma membrane by the action of DMSO. Scanning electron microscopy revealed that many filopodelike projections formed on the surface of cells treated with 5% DMSO for 5 min. Interestingly, the projections were formed on a restricted portion of the cell surface. The phagokinetic track technique of Albrecht-Buehler (1977, Cell, 11: 395-404) showed that the projection region corresponded to the anterior part of a migrating cell. The possible relationship between the DMSO-induced projection region on the cell surface and intracellular organization of cell organelles was investigated using serial thin sections. The DMSO-induced projections contained arrays of microfilaments; and the microtubule organizing center (MTOC), nucleus, and vesicular structure were usually located in this order from the anterior end of the cell. The indirect immunofluorescent study using monoclonal anti-alpha-tubulin antibody was performed with a new fixation technique, which greatly improved the phase as well as immunofluorescent microscopy. It was verified that the intracellular positioning of the MTOC and nucleus had significant correlation with the cell polarity. The results show that DMSO is a powerful tool with which to manipulate the cellular microfilaments and to make visible the differentiation in the cortex layer, which apparently is relevant to the intracellular positioning of cell organelles and cell polarity.

Cell Membrane↗
PubMed1983

Hyperproduction of adenovirus type 12 E1B gene product in monkey cells, using a simian virus 40 vector.

Simian virus 40 recombinant DNAs carrying the adenovirus type 12 E1B gene were constructed, propagated, and packaged in monkey cells. Monkey cells infected with the resulting virus stocks hyperproduced the E1B gene products in more than 80% of the cells as revealed by immunofluorescence. The products were distributed in both the nuclei and the cytoplasm, and a condensed form of fleck structure was observed in the cytoplasm. Polyacrylamide gel electrophoresis of the cell extracts and their immunoprecipitates detected the E1B-coded 19,000-molecular-weight protein but not the 50,000-molecular-weight protein. The 19,000-molecular-weight protein and the simian virus 40 VP1 protein were synthesized in nearly equal amounts.

Adenoviruses, Human↗
PubMed1983

Expression of adenovirus type 12 early region 1 in KB cells transformed by recombinants containing the gene.

The adenovirus type 12 (Ad12) early region 1 (E1) gene was introduced into KB cells by using a dominant selection vector, pSV2-gpt, and over 80 Gpt+ KB cell clones were established. Three types of recombinant DNAs (gAE1A, gARC, and gABA) were constructed. They contained the AccI-H, EcoRI-C, and BamHI-A fragments, respectively, of Ad12 DNA in pSV2-gpt. Five of 50 (10%) gABA-transformed cell clones, 12 of 18 (67%) gAE1A-transformed cell clones, and 10 of 18 (56%) gARC-transformed cell clones complemented the growth of Ad5 dl312 (deletion in E1A) and were designated as Gpt+ Ad+ cell clones. In these cell clones at their early passages, recombinant genome sequences were detected in cellular DNA and were expressed. T antigen g (the E1A gene product) was detected by immunofluorescence. The Gpt+ Ad+ cell clones supported the growth of Ad5 deletion mutants in parallel with the expression of Ad12 E1A or E1A plus E1B genes. After infection of Gpt+ Ad+ cell clones with Ad5 dl312, the early genes of dl312 were efficiently transcribed, indicating the expression of the pre-early function of the Ad12 E1A gene. Two clones each from gAE1A-,gARC-, and gABA-transformed cells were subcultured for a long period to determine the stability of the transfecting DNAs. Subculture in a nonselective medium resulted in cells which lost the transfecting DNAs. Subculture in a selective medium resulted in the selection of cells which maintained the gpt gene expression but lost the Ad12 gene expression. These results indicate that the transfecting DNA is present in an unstable state in KB cells.

Adenoviruses, Human↗
PubMed1983

Chromophoric labeling of cannabinoids with 4-dimethylaminoazobenzene-4'-sulfonyl chloride.

A simple and sensitive assay for the cannabinoids is presented using a dabsylation procedure. Dabsyl derivatives of delta 9-tetrahydrocannabinol (delta 9-THC) and cannabinol (CBN) were prepared by reacting with 4-dimethylaminoazobenzene-4'-sulfonyl chloride (dabsyl chloride) in acetone in the presence of sodium carbonate-sodium bicarbonate buffer (pH 10). Crystalline dabsylcannabinoids gave intense absorption in the visible region. With these derivatives, analysis by thin-layer chromatography (TLC) and high performance liquid chromatography (HPLC) were tested. These techniques gave good separation and nanogram detection of dabsyl-THC and -CBN by using n-hexane-ethyl acetate-diethylamine (20:5:1) for TLC and MeOH--H2O (95:5) at 450 nm for HPLC.

Cannabinoids↗
PubMed1983

Mesenchymal hamartoma of the liver in infant: its roentgen diagnosis.

A case of mesenchymal hamartoma of the liver in an 18-month-old boy is reported. CT and US revealed a large, cystic tumour surrounded by a thick wall. In the lower portion of the tumour, a small solid component was recognized on CT. Angiography revealed marked arterial displacement, fine tumour vessels and a large radiolucent area. CT, US and angiography were helpful to make a diagnosis of mesenchymal hamartoma of the liver.

Angiography↗
PubMed1982

In vitro nucleation of microtubules from microtubule-organizing center prepared from cellular slime mold.

Nucleus associated bodies (NABs) were isolated from Dictyostelium discoideum or Dictyostelium mucoroides and their ability to nucleate microtubules in vitro was examined. NABs were localized at the tapered ends of the nuclei and released from lysed cells in complex with the nuclei. Microtubules radiating from the NAB could also be isolated with the complex under microtubule stabilizing conditions. The ultrastructure of the isolated NAB showed it to be composed of a core structure surrounded by an amorphous matrix. The ability of isolated NABs to nucleate microtubules in vitro was demonstrated by incubation with exogenous brain microtubule protein. Microtubule assembly was easily visualized by dark-field or immunofluorescence microscopy. Polymerization of microtubules seemed to be initiated not from the core structure but from the surrounding matrix. The number of microtubules polymerized from the NAB was directly counted in whole-mount preparations by electron microscopy, which provided a quantitative assay for the NAB activity. The nucleating activity of NAB was quite unstable and its half-life was calculated as about 5 hours. The activity was sensitive to protease digestion and was also temperature sensitive but could be stabilized by addition of glycerol or storage at - 80 degrees C or in liquid nitrogen. These characteristics are analogous to those of the centrosomes in cultured mammalian cells and a possible explanation of their similarity is discussed.

Cell Fractionation↗
PubMed1982

Effect of gonadotropins, steroids and culture media on bovine oocyte maturation in vitro.

The present experiment was to investigate the effect of gonadotropins (LH and hCG), steroids (estradiol and progesterone) and culture media (TCM 199, Ham-F-12, BMOC-3 and modified KRB) on in vitro maturation of cumulus-enclosed bovine oocytes. Oocytes isolated from follicles of </=5 mm in diameter were cultured for 27 to 30 hours. Dosages of gonadotropins were 10 mug/ml and 2 i.u./ml for LH and hCG, respectively, and that of steroids was 1 mug/ml for both estradiol and progesterone. The study showed that the proportion of matured (metaphase II and a polar body formation) oocytes was significantly affected by the presence or absence of steroids (P < 0.01), but not by the gonadotropins and the media. Estradiol, but not progesterone, significantly increased the rate of oocyte maturation when compared with that of no steroid treatment (47.8%, 61.7% and 55.2% for none, estradiol and progesterone, respectively). These results indicate that steroids, especially estradiol, were the most important factor for bovine oocyte maturation rather than gonadotropins and culture media, and that the effect of the hormones differed in the media tested.

Journal Article↗
PubMed1982

A new method of evaluating the postimperative negative variation (PINV).

The PINV has been widely applied to psychiatric investigations in which the definition of its abnormality depended on the visual evaluation without giving attention to its wave form. We have developed a new method of evaluating the PINV by applying the exponential regression correlation to raw and smoothed PINVs for the amplitude of the 15 points at the intervals of 70 msec. The significant correlation between the PINV wave form and the exponential curve was observed both in the raw and smoothed PINVs. Coefficient A was positively correlated with the PINV duration quantified by a visual measurement. Coefficients A and B had the negative and positive correlation with the CNV magnitude. The findings obtained in 28 healthy subjects suggest that the PINV may correlate to the CNV magnitude.

Adult↗
PubMed1982

Melancholia and excessive CNV recovery after nonresponse condition.

The effect of nonresponse on the Cz-CNV recovery was studied in 13 depressed patients on the hypothesis that the excessive CNV recovery had a relation to melancholia. The depressed patients had a significantly small Control CNV area before the nonresponse as compared with a matched group of healthy controls. The dysthymic patients by the Maudsley Personality Inventory (MPI) showed a significantly smaller Control CNV area than the non-dysthymic patients. Melancholic patients who met the DSM-III criteria had a significantly longer control reaction time and more excessive CNV recovery than the non-melancholics. The excessive CNV recovery is useful as a predictor for melancholia, and the motivational facilitation and preparatory motor inhibition underlying the psychomotor retardation and hence "endogenous process" were discussed.

Adult↗
PubMed1982

Acute effects of ethanol and acetaldehyde on plasma phosphate level.

Oral administration of ethanol in a dose of 65 mmol kg-1 produced marked change of plasma phosphate level in rabbits. Hypophosphataemia was observed for the first 2 h after administration followed by significant increase of plasma phosphate at 5 h. Hypophosphataemia did not appear when ethanol was given to the rabbits pretreated with pyrazole. When animals were injected with disulfiram in advance, the duration of hyperphosphataemia due to ethanol was prolonged. Administration of acetaldehyde at a dose of 1.5 mmol kg-1 produced hyperphosphataemia. In this study, plasma phosphate was not associated with change in calcium level. These results suggest that the hypophosphataemia observed was related to the metabolic process of ethanol utilizing alcohol dehydrogenase, and that acetaldehyde, a metabolite of ethanol, might induce the hyperphosphataemia in the animals.

Acetaldehyde↗
PubMed1982

Dependence of tumor-forming capacities of cells transformed by recombinants between adenovirus types 5 and 12 on expression of early region 1.

Recombinants between an adenovirus type 5 (Ad5) deletion mutant and the Ad12 DNA fragment containing early region 1 (E1) were isolated from cells cotransfected with the EcoRI-C fragment of Ad12 DNA and Ad5 dl312 (deletion in E1A) DNA (rcA) and from cells cotransfected with the SalI-C fragment of Ad12 DNA and Ad5 dl312 DNA (rcB). No recombinant was isolated from cells cotransfected with Ad5 dl313 (deletion in E1B) DNA and restriction fragments of Ad12 DNA. Both rcA and rcB are defective and able to replicate in human embryo kidney (HEK) and KB cells with complementation by dl312. Both rcA and rcB formed Ad12 T antigen g, but not T antigen f, in infected HEK and KB cells. In rcA- and rcB-infected cells, Ad5 E1B and Ad12 E1A genes are transcribed. Heteroduplex and size analyses of rcA-1 or rcB-1 DNA fragments hybridized with Ad12 DNA revealed that rcA-1 DNA has a deletion between 5 and 15 map units with an insertion of a portion of Ad12 DNA (10%) and that rcB-1 DNA has a deletion between 70 and 80 map units with an insertion of a portion of Ad12 DNA (10%). The transformed cell lines, RCAY and RCBY, were established after infection of rat 3Y1 cells with rcA and rcB, respectively. Both Ad5 and Ad12 DNA sequences are contained in these cells. In RCAY cells, Ad12 T antigen g is detected, but Ad12 T antigen f is not. In RCBY cells, both Ad12 T antigen g and f are detected. Only the Ad12 E1A gene is transcribed in RCAY cells, whereas Ad5 E1B, Ad12 E1A, and Ad12 E1B genes are transcribed in RCBY cells. In soft-agar cultures, RCBY cells form large colonies, whereas RCAY cells form only tiny colonies. RCBY cells form tumors as efficiently as 12WY cells in transplanted rats. RCAY cells formed tumors inefficiently. Ad5-transformed 5WY cells do not form tumors. These observations indicate that the efficient tumor formation by RCBY cells is dependent on the expression of the Ad12 E1A and E1B genes, whereas the inefficient tumor formation by RCAY cells is due to the expression of only the Ad12 E1A gene.

Adenoviruses, Human↗