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Biomedical subjects

Y Fukui

Publications and source records attributed to Y Fukui.

At least 397 records · Page 22Linked to original sources
PubMed1989

Effects of sera, hormones and granulosa cells added to culture medium for in-vitro maturation, fertilization, cleavage and development of bovine oocytes.

Sera (fetal calf serum: FCS; and oestrous cow serum: ECS), hormones (2.5 FSH micrograms/ml + 5 micrograms LH/ml + 1 microgram oestradiol/ml) and granulosa cells (5 x 10(6)/ml) were added to culture medium to determine the frequencies of in-vitro maturation, fertilization, cleavage (2- to 8-cell) and development into blastocysts of bovine follicular oocytes. The maturation rates after 24 h in culture were not significantly different among the three factors tested (56-72%). The fertilization rates were significantly affected by serum type and the addition of granulosa cells. FCS gave significantly higher rates of fertilization (57-71%) than did ECS (34-52%), but the proportions of polyspermic fertilization were significantly higher in the former (8-19%) than in the latter (2-3%). The addition of hormones did not affect fertilization, cleavage and development. Neither type of serum affected cleavage and development. The highest rates of blastocyst formation were obtained when granulosa cells alone were added (FCS, 17%; ECS, 16%). The cell numbers of the blastocysts obtained were 100-150, similar to those of blastocysts developed in vivo. Transfer of 6 blastocysts to 3 cows resulted in 1 pregnancy. The present results indicate that the co-culture with granulosa cells is the most important factor for in-vitro fertilization to development into blastocysts of bovine oocytes matured in vitro.

Animals↗
PubMed1989

Effects of sera and steroid hormones on development of bovine oocytes matured and fertilized in vitro and co-cultured with bovine oviduct epithelial cells.

The present experiment was designed to identify possible effects of sera and steroid hormones added to a co-culture with bovine oviduct epithelial cells on embryonic development in vitro. Bovine oocytes were matured in vitro for 24 h and then fertilized in vitro using swim-up and heparin-treated, frozen-thawed spermatozoa. At 18 and 20 h after insemination, oocytes were cultured for 3 or 7 d in a co-culture system with bovine oviduct epithelial cells containing either fetal calf serum (FCS) or estrous cow serum (ECS) and one of six hormonal additions (none, 1 or 10 micrograms/ml estradiol [E]; 1 microgram/ml progesterone [P]; 1 microgram/ml E + P; and 10 micrograms/ml E + P). A total of 2,666 oocytes were cultured for 3 d and examined for cleavage. Of those, 2,280 oocytes were cultured up to 7 d for development to the late morula or blastocyst stage. Greatest cleavage rates for 2- to 8-cell and 8-cell stages were observed in FCS (71 and 24%) and ECS (66 and 23%) without steroid addition. For development into blastocysts, no serum effect was observed. Greatest rates for development into blastocysts were observed in FCS (14%) and ECS (16%) without steroid addition. These results indicate that addition of E and P at the doses and combinations tested did not enhance developmental capacity of in vitro fertilized bovine oocytes. Compared with FCS, ECS tended to increase cleavage rates and development into blastocysts.

Animals↗
PubMed1989

The effect of carbon dioxide on the changes in blood pH during hypoxia.

The change in blood pH during hypoxia was examined using rabbits. Rabbits were divided into 4 groups according to the test gases used (O2 5.1%: CO2 5.4%: N2 89.5%, O2 4.9%: N2 95.1%, O2 2.0%: CO2 5.6%: N2 92.4% and O2 1.9%: N2 98.1%). After an intravenous injection of urethane, the rabbit was fixed on its back on an operating table and the trachea was cannulated. Animals inhaled test gases through a cylindrical unidirectional valve box connected to the cannula. Blood samples were drawn from the catheterized femoral artery. The duration of exposure was 90 min. The animals in the CO2-added 5% O2 group survived the exposure, while only 2 animals survived in the CO2-free 5% O2 group. There was a difference of about 5 mmHg in the blood PO2 between both 5% O2 groups. There was also a marked difference in the time course of changes in the hydrogen ion [( H+]) concentrations between the groups. The [H+] in the CO2-free group began to increase after an initial fall and exceeded the value of the other group, reaching acidotic levels. The acidosis (metabolic) was considered to have been caused by the accumulation of lactic acid. Under the present experimental conditions the contribution of hypoxia to this acidosis was considered to be greater than that of hypocapnia. The animals in the 2% O2 groups died by the 11 min after the start of exposure. In the CO2-free group neither hypercapnia nor acidosis was observed during the exposure. The PO2 values at apnea were about 10 mmHg in both groups.

Administration, Inhalation↗
PubMed1989

Effect of ethanol on interaction of hypoxia and PCO2 levels in rabbits.

The following three kinds of hypoxia were imposed on rabbits: the inhalation of two hypoxic gas mixtures (O2 2%-CO2 5%-N2 93% [CO2-added hypoxic] and O2 2%-N2 98% [CO2-free hypoxic]) and asphyxiation by tracheal occlusion. Ethanol (1.5g/kg of body weight) was given intravenously 30 min before the start of the hypoxia. A period up to the onset of apnea, as well as death, was shorter in the ethanol-treated group than in the non-treated group. This respiratory depressing effect of ethanol was evident in the hypoxic gas inhalation group. There was no significant difference in the PO2 value when apnea occurred. The animals inhaling the CO2-free hypoxic gas mixture became apneic sooner than those inhaling the CO2-added hypoxic gas mixture. The respiratory depressing interaction between hypoxia and hypocapnia was increased by ethanol. Gasping was observed in about half of the animals after apnea of various lengths. The rate of gasping was affected by neither ethanol nor the way by which hypoxia was induced.

Animals↗
PubMed1989

Phosphatidylinositol kinase type I activity associates with various oncogene products.

We have assayed immunoprecipitates of several oncogene products from retrovirally infected chicken embryo fibroblasts (CEF) for phosphatidylinositol (PI) kinase activity. Immunoprecipitates of P68gag-ros, P130gag-tps, P47gag-crk, polyoma middle T (mT)-p60c-src complex, and mT-p62c-yrs complex exhibited PI kinase activity when assayed without detergents. This activity was sensitive to the nonionic detergent, Triton X-100, and the product was indistinguishable from phosphatidylinositol-3-phosphate, the product of kinase type I. Immunoprecipitates of p21Ha-ros protein did not contain any PI kinase type I activity. It has been suggested that an 81 kD protein phosphorylated in in vitro kinase assays of immunoprecipitates from mT-transformed rodent cells is responsible for the PI kinase type I activity seen in these immunoprecipitates. We have detected a chicken homologue of this 81 kD protein in immunoprecipitates of lysates from mT-transformed CEF. However, the chicken 81 kD protein sedimented more quickly than the PI kinase activity in sucrose gradients. In addition, the 81 kD protein was not detectable in protein kinase assays of immunoprecipitates of P68gag-ros or P130gag-fps. These results suggest that the 81 kD protein may not be the PI kinase.

1-Phosphatidylinositol 4-Kinase↗
PubMed1989

Parallel first-order and Michaelis-Menten elimination kinetics of ethanol. Respective role of alcohol dehydrogenase (ADH), non-ADH and first-order pathways.

Elimination kinetics of ethanol without (control group) and with pyrazole [alcohol dehydrogenase (ADH) inhibitor] pretreatment was studied with changing the i.v. dose amount to evaluate the respective role of ADH and non-ADH pathways in a rabbit. The moment analysis of the blood ethanol concentration-time curves showed that the normalized area under the blood ethanol concentration-time curve and the first moment increase with increasing dose amount in the control and pyrazole-pretreated groups. These increases suggested the capacity-limited elimination of ethanol through pyrazole-insensitive non-ADH pathways as well as through ADH pathway as pyrazole would fully block the oxidation of ethanol through ADH pathway. The simultaneous multiline fitting using time curves after five different doses also was attempted to determine the pharmacokinetic model by the application of minimum Akaike's information criterion estimation. Akaike's information criterion, consequently, showed the minimum for a two-compartment model with parallel first-order and Michaelis-Menten elimination kinetics. The computer analysis using this model yielded almost the same values of the volume of distribution and of the first-order elimination rate constant between both groups. The distribution of ethanol and the first-order elimination process were not influenced by pyrazole treatment. Km (0.57 mg/ml) of the pyrazole-pretreated group was higher than Km (0.03 mg/ml) of the control group. These results suggest that ADH pathway is readily saturated and non-ADH pathways are unsaturated over the wide range of concentration. The first-order process as well as non-ADH pathways are concluded to occupy the considerable part in the ethanol elimination at higher blood concentration.

Alcohol Dehydrogenase↗
PubMed1989

[Development of fuzzy blood pressure control system].

Fuzzy blood pressure control system has been developed to regulated blood pressure by using vaso-active drugs, which aims the medical treatment of the patient. Fuzzy logic is used to convert heuristic control rules as a physician's knowledge into an automatic control strategy. Usually, a physician decides infusion rate based on referring ideal pressure trajectory during control. To emulate such a physician's heuristic control, the controller refer an ideal pressure trajectory, for example, from the normal level to the target level. One of Inputs to the controller is a difference between real pressure and ideal pressure, where another is a rate of its pressure difference. An output from the controller is the change in drug infusion rate. Both of inputs and output are transformed into fuzzy sets, whose membership functions are predefined. Twenty one physician's control rules are converted into fuzzy control rules. The fuzzy control procedure consists of three steps: (1) computing both input values and finding their appropriate membership values (grade), (2) finding applied rules and applying fuzzy composition to infer weighted output fuzzy sets, (3) inferring the change in drug infusion rate by taking the center of mass of the membership function of output. Nine animal experiments are performed to evaluate the developed system by using rats with vasopressor drug. Angiotensin II Blood pressure was elevated from normal pressure (around 100 mmHg) to 150 mmHg. Satisfactory blood pressure control was realized on eight experiments and effectiveness of the controller was confirmed confirmed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Unique action of a modified weakly acidic uncoupler without an acidic group, methylated SF 6847, as an inhibitor of oxidative phosphorylation with no uncoupling activity: possible identity of uncoupler binding protein.

The potent weakly acidic uncoupler SF 6847 was modified by methylation of its phenolic OH group, and the effect of the resulting derivative, with no acid-dissociable group, on oxidative phosphorylation in rat liver mitochondria was examined. The methylated SF 6847 did not induce uncoupling at up to 40 microM, while SF 6847 uncoupled oxidative phosphorylation completely at about 20 nM, indicating that the acid-dissociable group is essential for uncoupling. The O-methylated SF 6847 at 20 microM did, however, inhibit state 3 respiration of mitochondria, although it did not inhibit electron-flow through the respiratory chain, ATPase activated by weakly acidic uncouplers or Pi-ATP exchange. At the same concentration, it also inhibited ATP synthesis in submitochondrial particles. These features are different from those of known inhibitors of oxidative phosphorylation. Thus, O-methylated SF 6847 is a unique inhibitor of oxidative phosphorylation. The possible identity of the uncoupler binding protein is discussed on the basis of these results.

Adenosine Triphosphate↗
PubMed1988

Gas chromatographic determination for forensic purposes of petroleum fuel inhaled just before fatal burning.

The determination of petroleum fuel in the blood of burned bodies was carried out by three different gas chromatographic procedures. Seven components of gasoline (isopentane, n-pentane, 2-methylpentane, benzene, 2-methylhexane, 3-methylhexane and toluene) and five of kerosene (xylene, C9H20, mesitylene, pseudocumene and C11H24) were chosen as indicators with a coefficient of variation of 5-24%. The methods were applied to four autopsy cases with a relatively low carboxyhaemoglobin (HbCO) content. When gasoline exposure had occurred, the blood concentrations determined were almost identical whatever the components selected. Great variations in the components determined were found after kerosene exposure, and hydrocarbons greater than or equal to C14 were hardly inhaled by the victims. A higher content of fuel in the left than in the right ventricular blood observed in the autopsy cases suggests fuel inhalation just before death. The same phenomenon was also observed in the content of blood HbCO. Determinations of petroleum fuel and HbCO in both the right and left ventricular blood would be useful for the forensic diagnosis on burned bodies with a low HbCO content.

Aged↗
PubMed1988

Strain differences in teratogenic susceptibility to trypan blue between WM and BDIX rat strains.

Female rats of WM (Wistar-Mishima)/Nem strain were mated with WM/Nem (group W) or BDIX/Nem males (group WB), and BDIX/Nem females were mated with BDIX/Nem (group B) or WM/Nem males (group BW). On day 8 of gestation, pregnant females were treated intraperitoneally with 1% aqueous solution of trypan blue at a dose of between 20 and 120 mg/kg of body weight. On day 20 of gestation, fetuses were examined for external, visceral, and skeletal malformations. In group W, fetal mortality increased dose dependently at doses higher than 20 mg/kg, and incidences of external, visceral, and skeletal malformations were significantly higher than control at doses of 30 mg/kg and more. In group B, fetal mortality and the incidence of external malformations were significantly higher than control only in the group treated with 120 mg/kg, and no significant increase of visceral and skeletal malformations was shown. It was confirmed that BDIX strain is much more resistant to trypan blue teratogenicity than WM strain. In group BW, nearly the same teratogenic effects were shown as in group W in terms of fetal mortality and incidence of malformations. However, in group WB, teratogenic effects were not so remarkable as in group BW, suggesting patroclinous effects in teratogenic susceptibility to trypan blue. In group BW, sex differences in teratogenic susceptibility were found; male fetuses were more susceptible to trypan blue than females.

Animals↗
PubMed1988

Isolation and characterization of Schizosaccharomyces pombe mutants phenotypically similar to ras1-.

We isolated mutants of Schizosaccharomyces pombe which have deformed cell morphology, are deficient in conjugation and poor in sporulation. This phenotype is characteristic of the ras1 defective mutant previously identified. Tests of the mutants for allelism using cell fusion showed that they define five complementation groups, one of which is ras1 itself. The others are named ral1 through ral4 (ras like). Mutants in ral3 or ral4 conjugate at a very low frequency, while the others apparently do not conjugate at all. Plasmid clones complementing ral1, ral2 or ral3, which apparently carry the respective gene, were isolated from S. pombe genomic libraries. Multiple copies of either the ral2 or the ral3 gene could partially restore mating ability in ral1- strains. Multiple copies of the ras1 gene could partially restore mating ability in ral1- and ral2- strains. These results suggest that the ral1, ral2 and ras1 genes may function in a common pathway in that order. The ral3 gene may influence this pathway. Analysis of these gene products will aid identification of factors which interact with Ras proteins.

Alleles↗
PubMed1988

In vitro culture of sheep oocytes matured and fertilized in vitro.

Sheep oocytes that matured and fertilized in vitro were cultured to evaluate their cleavage to the 8- to 16- cell stage and further development in five different media as follows: 1) CPMW (TCM199 + 20% ewe serum + 0.4% BSA), 2) Ham's F-10 + 10% ewe serum, 3) Brinster's pyruvate medium + 0.1% glucose (BPM-G), 4) co-culture with sheep oviduct epithelial cells in TCM199 + 10% fetal calf serum, and 5) co-culture with sheep granulosa cells in the same medium as 4. The culture duration was 4 or 7 d for 8- to 16-cell or further development. The proportions of 8- to 16-cell eggs were 1) 16% (8 49 ), 2) 25% (12 49 ), 3) 52% (58 112 ), 4) 63% (105 167 ) and 5) 45% (27 60 ). The co-culture with sheep oviduct cells resulted in a significantly (P < 0.05) higher rate of cleavage than the other media, except BPM-G. The proportion of noncompacted morula (35%, 24 68 ) was also significantly (P < 0.05) higher in the co-culture of sheep oviduct cells than the other media. The 8- to 16-cell eggs produced by BPM-G (n=38) and the co-culture with sheep oviduct cells (n=42) were transferred into the uterus of recipient ewes, but no elongated blastocysts were obtained 13 d later. On the other hand, 8 out of 55 one-cell eggs (15 to 18 h after in vitro insemination) transferred to the oviduct of recipient ewes were elongated blastocysts (24% of 34 recovered eggs). The data show that the co-culture of in vitro fertilized eggs with sheep oviduct epithelial cells could support development of 8- to 16-cell embryos or early morula, but their viability is still questionable.

Journal Article↗
PubMed1988

Effects of two treatments on semen from different bulls on in vitro fertilization results of bovine oocytes.

The semen of six different bulls was used to examine the effects of treatment with caffeine or caffeine plus Ca-ionophre on in vitro fertilization, cleavage and development into morulae of in vitro matured bovine oocytes. In vitro fertilization results (formation of both pronuclei, cleavage and development to >or= four-cell stage were significantly (P<0.01) higher using caffeine plus Ca-ionophre than those using only caffeine. The rates of fertilization and first cleavage were only slightly variable among the bulls. However, the present data showed significant variability in formation of both pronuclei (36 to 75%) of fertilized ova and development to the >or=4 cell stage (39 to 71%) by different bulls. Development into morulae of ova recovered from the rabbit oviduct did not show any significant differences in relation to sperm treatments or individual bulls.

Journal Article↗
PubMed1988

Increased plasma free fatty acid and triglyceride levels after single administration of toluene in rabbits.

Changes of plasma lipids (triglyceride, TG: total cholesterol, Cho; and phospholipids, PL), free fatty acid (FFA), and blood glucose (BG) were studied in male rabbits after toluene administration (0.5 g/kg per os). Hypertriglyceridemia was observed at and after 2 h. Plasma FFA and BG were elevated temporarily during the early stage and lowered gradually thereafter. Initially, plasma Cho and PL were virtually unchanged, but the Cho level increased slowly after 6 h. The hypertriglyceridemia observed may have some adverse effects on heart function.

Animals↗
PubMed1988

Lymphokine-activated killer (LAK) activity against autologous malignant tumors of the skin.

When peripheral blood mononuclear cells (PBMC) cultured with interleukin 2 (IL-2) develop the ability to lyse fresh tumor cells, such activity is referred to as lymphokine-activated killer (LAK) activity. In this paper, we examined LAK activity against the autologous skin tumors, malignant melanoma (MM), squamous cell carcinoma (SCC), and basal cell epithelioma (BCE), which have distinct clinical characteristics. Similar levels of LAK activity against Daudi 1A4, an NK resistant cell line were significantly obtained from all cancer patients. However, different levels of LAK activity against autologous tumor cells were found from three kinds of cancer patients using mixtures of autologous tumors and LAK. The levels of cytotoxicity were 29.8 +/- 7.0% in five MM, 15.9 +/- 4.9% in seven SCC, and 4.0 +/- 2.3% in five BCE patients at an effector/target ratio of 50/1. Allogeneic MM targets were lysed by LAK from all three types of cancer patients at similar levels, implying that LAK is not restricted to major histocompatibility complex (MHC) antigens. These results indicate that the levels of autologous LAK activity were significantly associated with the magnitude of clinical malignancy of the tumor targets, and suggested the selective lysis of tumor targets by LAK. NK activity of cancer patients bearing tumors was also examined prior to therapy. The levels of NK activity observed in MM patients were considerably lower than those in two other cancer patients.

Adult↗
PubMed1988

Detection of gangliosides as N-glycolylneuraminic acid-specific tumor-associated Hanganutziu-Deicher antigen in human retinoblastoma cells.

Gangliosides were shown to bear the tumor-associated N-glycolylneuraminic acid (NeuGc)-specific Hanganutziu-Deicher (HD) antigen expressed in human retinoblastoma cells. HD antigenic gangliosides were detected by thin-layer chromatography/enzyme-immunostaining using affinity-purified chicken antibody against GM3 containing NeuGc and horseradish peroxidase-conjugated anti-chicken IgG. One to four species of the antigenic gangliosides were detected from all of 4 cell lines, Y79, WERI-Rb1, TOTL1, and YK, as well as freshly cultured retinoblastoma cells and isolated tumor tissue. All cases contained GM3(NeuGc) as an HD antigen. No HD antigenic ganglioside was detected in normal retinal tissues by the same procedure.

Antigens, Heterophile↗
PubMed1988

Amplification of the structurally and functionally altered epidermal growth factor receptor gene (c-erbB) in human brain tumors.

By using Southern blot analysis, we found that in two cases of human glioblastoma multiforme, cells carried amplified c-erbB genes which bore short deletion mutations within the ligand-binding domain of the epidermal growth factor (EGF) receptor. The products of these mutated c-erbB genes were about 30 kilodalton (kDa) smaller than the normal 170-kDa EGF receptor, and the tumor cell membrane fractions containing the 140-kDa abnormal EGF receptor showed a significant elevation of tyrosine kinase activity without its ligand. In view of the similarity to the activated viral and cellular erbB genes in the avian system, these mutated and overexpressed EGF receptors might play a role in the onset or development of human glioblastoma cells.

Animals↗