[Response of exposed pulp to composite resin in germ-free rat molars].
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Animals↗
Biomedical subjects
Y Okamura
Publications and source records attributed to Y Okamura.
[Establishment of primary culture of cells derived from uterine leiomyoma].
To evaluate the effects of drugs on uterine leiomyoma and to clarify the histogenesis of uterine leiomyoma, we studied the establishment of primary culture of cells derived from uterine leiomyoma. Myoma tissues were cut into small pieces and suspended in trypsin. The primary cell culture, as a monolayer, was able to be passaged four times. To confirm that the cultured cells were derived from the myoma, the cells were stained with desmin by the enzyme-labelled antibody method. The cultured cells derived from uterine muscle cells were similarly treated with desmin staining. The results confirmed the morphological similarity between the two groups of cells. To confirm that the cultured cells were myoblast, fibroblasts derived from the myoma were cultured selectively. The cells that resembled myoblasts, morphologically appeared spindle-shaped. The cells that resembled fibroblast, appeared polygonal and extended over the bottom of the culture flask. The growth curve of cultured myoma cells (8.0 X 10(4) cells in 0.2 ml) in Petri dishes (60 X 15 mm) revealed logarithmic proliferation after about 5 days. The colony formation of cultured myoma cells (2 X 10(5) cells in 0.2 ml) in culture flasks (25 cm2 in the area of the base) morphologically appeared to have an irregular border and had many independent scattered cells around the colony when the medium was renewed twice a week. The myoma cells (8.8 X 10(4) in 0.2 ml) in the logarithmic phase were cultured, fixed after 12 days when the medium was not renewed, and stained with crystal-violet. Morphologically, the colony had a comparatively regular border, was round, and each cell in the colony was epithelioid. The plating efficiency was 0.07%.(ABSTRACT TRUNCATED AT 250 WORDS)
Cells, Cultured↗
[Human placental glutathione transport mechanism].
The placental transport mechanism of glutathione (GSH) was investigated using microvillous membrane vesicles prepared from human term placenta. Using (3H-glycine)-labeled-GSH, it was clarified that GSH in the extravesicular compartment of placental microvillous membranes was rapidly degraded by gamma-GTP (gamma-glutamyltranspeptidase) and resulting amino acid, and 3H-labeled-glycine was actively transported via a sodium cotransport system. AT-125 treated microvillous membrane vesicles almost entirely lost its gamma-GTP activity, and showed intact GSH transport. Using AT-125 treated microvillous membrane vesicles, it was revealed that GSH was transported across the microvillous membrane as an anion via a membrane potential-dependent mechanism. These results indicated that gamma-GTP which existed in microvillous membrane played a role in GSH metabolism and that intracellular GSH was translocated out of the syncythiotrophoblast cell into the maternal blood space via a specific carrier in microvillous membrane because the GSH concentration was higher in intracellular than extracellular and extracellular membrane potential was positively charged.
Biological Transport, Active↗
[The study on human placental DHA-S transport mechanism (using placental microvillous membrane vesicles)].
To investigate the placental DHA-S (dehydroepiandrosterone sulfate) transport mechanism, the uptake of DHA-S into microvillous membrane vesicles prepared from human term placenta was studied using the rapid filtration technique. 1. The uptake of DHA-S into microvillous membrane vesicles was not dependent on both Na+ electrochemical gradient and membrane potential difference. 2. The uptake of DHA-S into microvillous membrane vesicles was dependent on temperature. The initial uptake rate of DHA-S at 37 degrees C was three times as great as at 4 degrees C. 3. The initial rate of DHA-S transport exhibited saturation kinetics with respect to the DHA-S concentration; an apparent Km of 0.067 mM and Vmax of 1.01 nmol/mg protein/20 sec were calculated. 4. The uptake of DHA-S into microvillous membrane vesicles was inhibited by DHA, but not by estriol. These results indicated that placental DHA-S transport was carrier mediated and a passive one.
Biological Transport↗
[The effect of inorganic mercury on placental amino acid transport using microvillous membrane vesicles].
To study the toxic effect of inorganic mercury on the placenta we elucidate the effect of inorganic mercury on placental amino acid transport using microvillous membrane vesicles isolated from human normal full term placenta. The transport of amino acids into microvillous membrane vesicles was studied by a rapid filtration technique using a millipore filter. The transport of L-alanine across placental microvillous membrane was Na+ electrochemical gradient dependent and 0.1 mM inorganic mercury inhibited 77% of this Na+ dependent L-alanine transport and 1 mM inorganic mercury inhibits 90% of this Na+ dependent L-alanine transport. The transport of L-lysine across microvillous membrane vesicles was sodium independent and 0.1 mM inorganic mercury inhibited 34% of this transport and 1 mM inorganic mercury inhibited 50% of this transport. These results indicated that one of the toxic effects of inorganic mercury on placenta-fetus unit was the inhibition of placental nutrient transport.
Alanine↗
[Study on human placental beta-alanine and taurine transport mechanism (using microvillous membrane vesicles].
Using microvillous membrane vesicles prepared from human full term placenta, we studied the placental beta-amino acid transport mechanism. The transport of amino acids into microvillous membrane vesicles was studied by a filtration technique using a millipore filter. The uptake of beta-alanine into microvillous membrane vesicles was dependent on Na+ electrochemical gradient (extravesicular greater than intravesicular). The initial rate of this Na+ gradient dependent beta-alanine transport exhibited saturation kinetics with respect to the beta-alanine concentration: an apparent Km of 0.24 mM and Vmax of 46 pmol/mg protein/20 sec were calculated. Taurine inhibited beta-alanine uptake into microvillous membrane vesicles, but on the other hand L-alanine didn't inhibit this beta-alanine uptake. The L-alanine uptake into microvillous membrane vesicles was Na+ electrochemical gradient dependent and the initial rate of this Na+ dependent L-alanine uptake into vesicles was faster than the uptake of Na+ itself into vesicles. On the other hand, the initial rate of Na+ dependent beta-alanine and taurine uptake into vesicles was slower than the uptake of Na+ itself into vesicles. These results indicated that there existed a beta-amino acid specific transport system in human placental microvillous membrane, and placental taurine transport was carried out by this system. And it was also indicated that this placental beta-amino acid transport mechanism is quite different from that of L-alanine.
Alanine↗
[A case report of an advanced ovarian cancer (stage T4) treated with a total amount of 1815 mg of CDDP].
A case is reported of a 34-year-old female who complained of an adult head-sized tumor in the pelvis and was treated with a high dose of CDDP. On the first look operation, it was found impossible to perform reduction surgery because of severe adhesion and peritonitis carcinomatosa. To introduce a high concentration of an anti-cancer agent into the tumor tissue, we administered CDDP intravenously, intraperitoneally and intra-arterially to the total amount of 1815 mg. After this high dose combination chemotherapy, a second look operation was performed and the tumor was seen to be reduced. The patient is now in remission and is being followed up as an out-patient. The clinical course of this case had decided for us which route would be the most effective and would have the least side effect, also, how much of a dosage can be administered as a maximum adoptive dose of an anti-cancer agent, and what kind of a second-line chemotherapy is effective against resistant cancer cells.
Adult↗
[The role of glutathione on placental amino acid transport (using microvillous membrane vesicles)].
To elucidate the role of glutathione (GSH) on placental amino acid transport, we investigated L-lysine transport using microvillous membrane vesicles prepared from full term human placenta. 1. The transport of L-lysine into microvillous membrane vesicles was not affected by glutathione. 2. The transport of L-lysine into microvillous membrane vesicles was inhibited by inorganic mercury (Hg2+), and 0.1mM Hg2+ inhibited 34% of this transport and 1mM Hg2+ inhibited 50%. 3. The transport of L-lysine inhibited by Hg2+ was almost completely restored when glutathione was added simultaneously. These results indicated that glutathione defended the inhibitory action of inorganic mercury on L-lysine transport across microvillous membrane.
Amino Acids↗
[Staining of leiomyoma tissue and myoma-derived cultured cells by the enzyme-labelled antibody method--using desmin, vimentin and fibronectin].
To elucidate the pathogenesis of leiomyoma, myoma tissues and myoma-derived cultured cells were stained with desmin, vimentin and fibronectin by the enzyme-labelled antibody method. Desmin staining was positive in both the myoma tissues and the cultured cells. Vimentin staining was positive in endothelial cells in the blood vessels of the myoma and in the cultured cells. Fibronectin staining was positive in both the myoma and cultured cells. These findings demonstrate that the leiomyoma was derived from myogenic cells. It was suggested that the myoma-derived cultured cells were myofibroblasts with myoblasts and fibroblasts, and had the ability to differentiate. It appears that the original cells of the myoma differentiate to myofibroblasts, which then differentiate into myoblasts and fibroblasts.
Adult↗
[The changes in EPH gestosis placental amino acid transport activity (using human placental microvillous membrane vesicles)].
To elucidate the change in EPH gestosis placental amino acid transport activity, we investigated the uptake of L-alanine into microvillous membrane vesicles prepared from EPH gestosis placenta and from normal placenta by using a rapid filtration technique. 1. Alkaline phosphatase (ALP) was the marker enzyme of microvillous membrane vesicles (MMV). The ALP activity of mild EPH gestosis placental MMV didn't differ from that of normal placental MMV. On the other hand, the ALP activity of severe EPH gestosis placental MMV decreased compared to that of normal placental MMV. 2. The uptake of L-alanine into human placental MMV was dependent on the Na+ electrochemical gradient, so the transport across human placental MMV was a secondarily active one. The L-alanine transport activity of mild EPH gestosis placental MMV didn't differ from that of normal placental MMV. On the other hand, the L-alanine transport activity of severe EPH gestosis placental MMV decreased prominently compared to that of normal placental MMV.
Alanine↗
[Cold constitution: analysis of the questionnaire].
"Hie-sho" means having a cold constitution. The authors studied basic data for cold constitution with a questionnaire and with the Cornell Medical Index. The subjects were 318 females, and the average age was 26.4 +/- 6.1 (Mean +/- SD) and the range was 20 approximately 51 years of age. The results were as follows: (1) 38.7% of the subjects had been conscious of having a cold constitution, and it was recognized that the subjects with a cold constitution are found not only in the climacterium but also in the young. (2) The average age of onset of cold constitution was 19.3 +/- 5.1 years old. (3) The parts of the body that usually felt cold were first the feet and then the hands. (4) In winter or at bedtime the cold sensation was increased. And it is characteristic that the sensation was changed by weather or by body condition, especially by the stimulation of coldness. (5) Most subjects with a cold constitution took care in their daily life, but very few of them were treated. (6) Many other physical symptoms were recognized in subjects with a cold constitution, and their CMI scores were high. Therefore these data may be regarded as indicating that most cases of cold constitution are a partial symptom of vegetative disturbance (autonomic nerve dysfunction). (7) Most of the mothers of subjects with a cold constitution also had a cold constitution, which suggests the involvement of a hereditary factor concerning in the development of a cold constitution.
Adult↗
[The effect of antibiotics (gentamicin) on placental amino acid transport activity (using human placental microvillous membrane vesicles)].
In order to elucidate the effect of antibiotics (gentamicin) on placental amino acids transport, we investigated L-alanine transport using microvillous membrane vesicles prepared from full-term human placental by a rapid filtration technique. 1. The active transport of L-alanine into microvillous membrane vesicles was dependent on Na+ electrochemical gradient (extravesicular greater than intravesicular). And the double reciprocal plot of this Na+ dependent initial uptake rate versus L-alanine concentration exhibited an apparent Km of 0.79 + 0.23mM and a Vmax of 3.56 + 0.70n mol/mg protein/20 sec. 2. Gentamicin did not affect the Km value of this Na+ dependent L-alanine transport kinetics (0.77 + 0.19 mM [lmM gentamicin], 0.79 + 0.21mM [10mM gentamicin]). On the other hand, gentamicin apparently decreased, the Vmax value of this transport kinetics (1.99 + 0.48n mol/mg protein/20 sec [1mM gentamicin], 1.12 + 0.32n mol/mg protein/20 sec [10mM gentamicin]).
Alanine↗
[Perrault's syndrome: familial gonadal dysgenesis with sensorineural deafness].
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Adult↗
[Nonenzymatic glucosylation of human placental trophoblast basement membrane collagen (relation to diabetic placenta pathology)].
Nonenzymatic glucosylation is a reaction in which glucose binds nonenzymatically to hemoglobin, serum protein and glomerular basement membrane collagen etc. It has been thought that nonenzymatic glucosylation results in functional and chemical changes in those substances (hemoglobin etc) and contributes to the pathological changes in diabetes mellitus. This time we investigated whether nonenzymatic glucosylation occurred in human placental trophoblast basement (TrBM) collagen or not. The ability of glucose to interact with TrBM collagen (nonenzymatic glucosylation of TrBM collagen) was examined by incubating TrBM collagen with 3H-D-glucose in vitro. As a result it was shown that nonenzymatic glucosylation occurred in TrBM collagen and nonenzymatic glucosylation of TrBM collagen depended on the glucose concentration, reaction time and reaction temperature. These results indicate that possibly hyperglycemia, via nonenzymatic glucosylation modifies the function and chemistry of TrBM collagen and is related to the placental pathological changes in diabetic pregnancy.
Adult↗
[Intraperitoneal high-dose cisplatinum chemotherapy (CDDP-ip) in patients with carcinomatous peritonitis].
Immediately after CDDP-ip, the level of free Pt in ascites reached nearly 100 micrograms/ml, and the AUC (area under the curve) for ascites was 20-140 times greater than that for serum. The free Pt in serum following CDDP-ip administration was detected for several hours, and interestingly, the AUC for serum after ip therapy was 0.4-2.2 times greater than that after iv therapy. As a result, free Pt was found to act on cancer cells in the abdominal cavity directly at a high concentration. At the same time, the possibility of an antitumor effect from the vascular side of the tumor was also suggested. On the other hand, cases of ovarian cancer had various levels of peritoneal clearance (CLp), which depended on the severity of their carcinomatous peritonitis. The CLp had a great influence on the peak plasma concentration and on the AUC of free Pt in serum. In particular, the peak plasma concentration produced by CDDP-ip was 40-80% of the plasma concentration produced by CDDP-iv. These findings indicate that high-dose CDDP-ip is possibly effective and useful for advanced ovarian cancer, producing only very mild side effects.
Ascitic Fluid↗
The study of placental L-ascorbate (vitamin C) transport mechanism (using microvillous membrane vesicles).
To investigate the placental L-ascorbate (Vitamin C) transport mechanism, the uptake of L-ascorbate into microvillous membrane vesicles prepared from human term placenta was studied using the rapid filtration technique. The uptake of L-ascorbate into microvillous membrane vesicles was osmotically sensitive. This finding indicated that the uptake of L-ascorbate into microvillous membrane vesicles represented transport into the vesicles. The uptake of L-ascorbate into microvillous membrane vesicles was not dependent on sodium electrochemical gradient. The initial rate of uptake was not changed when the intravesicular space was rendered electrically more negative by membrane diffusion potential induced by the use of highly permeant anions. The initial rate of L-ascorbate transport exhibited saturation kinetics with respect to the L-ascorbate concentration; an apparent Km of 1.33 mM and Vmax of 47p mol/mg protein/20 sec was calculated. The uptake of L-ascorbate into microvillous membrane vesicles was competitively inhibited by D-isoascorbate. These results indicated that transport of L-ascorbate across the placental microvillous membrane vesicles was carrier mediated and was passive transport.
Ascorbic Acid↗
[D-glucose reabsorption activity of fetal kidney (by using brush border membrane vesicles of proximal tubule)].
By using brush border membrane vesicles prepared from human mid gestational fetal kidney (cortex), the D-glucose transport mechanism and its developmental pattern was studied by the rapid filtration technique. 1. The transport of D-glucose into brush border membrane vesicles prepared from fetal kidney (18 gestational weeks) did not show a great degree of Na+ gradient (extravesicular greater than intravesicular) dependence. 2. The transport of D-glucose into brush border membrane vesicles prepared from fetal kidney (22 gestational weeks) depended on Na+ gradient. This Na+ dependent D-glucose transport showed overshoot and markedly increased when the intravesicular space was rendered electrically more negative with a membrane potential induced by highly permeant anions or by the K+ diffusion membrane potential via valinomycin. These results indicated that the Na+ dependent active D-glucose reabsorption system in the proximal tubules of kidney might be formed between 18 and 22 gestational weeks and the D-glucose reabsorption mechanism of kidney at 22 gestational weeks was Na+ cotransport dependent on membrane potential as in adult ones.
Absorption↗
[Absorption of L-alanine in small intestine of human fetus (by using brush border membrane vesicles of jejunum)].
By using brush border membrane vesicles prepared from human mid gestational fetal intestine (jejunum), the intestinal L-alanine transport mechanism was studied using a rapid filtration technique. The uptake of L-alanine by the vesicles was osmotically sensitive. This finding indicated that the uptake of L-alanine by the vesicles represented transport into the vesicles. A Na+ electrochemical gradient (extravesicular greater than intravesicular) stimulated the initial rate of L-alanine uptake, and Na+ dependent uptake of L-alanine into vesicles showed a typical overshoot phenomenon. This overshoot and the initial rate of uptake markedly increased when the intravesicular space was rendered electrically more negative by membrane diffusion potentials induced by with highly permeant anions. A similar stimulation of L-alanine uptake was observed, when membrane potential (inside negative) was imposed by K+ diffusion potentials via valinomycin. These results indicated that an Na+ dependent uptake of L-alanine into the brush border membrane vesicles was dependent on the electrical potential difference of the membrane. The initial rate of L-alanine transport exhibited saturation kinetics with respect to the L-alanine concentration; an apparent Km of Km1; 1.6mM, Km2; 14.1mM, and Vmax of Vmax1; 4.0n mol/mg protein/20sec, Vmax2; 8.2n mol/mg protein/20 sec were calculated. From these results mid gestational fetal intestine (jejunum) already has an L-alanine absorption system which is comparable to the adult one's.
Alanine↗