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Biomedical subjects

K Tanabe

Publications and source records attributed to K Tanabe.

At least 541 records · Page 30Linked to original sources

Inhibin activity of preovulatory follicles of gonadotropin-treated and untreated women.

In order to determine whether treatment of endocrinologically normal women with human menopausal gonadotropin (hMG) and human chorionic gonadotropin (hCG) led to alternations in follicular fluid inhibin activity and steroids, both inhibin activity and steroid levels were measured in follicular fluid of 10 follicles of 9 women treated with hMG/hCG and in follicular fluid of 15 preovulatory follicles of 15 untreated women. The women were given 150 IU hMG daily from day 3 until the serum estrogen levels were greater than 300 pg/ml, at which time hMG was discontinued and 10,000 IU hCG was given 50 hours later. Follicular fluid was aspirated from all visible follicles 36 hours after hCG. Women treated with hMG/hCG had multiple (two to five) follicles containing 1.5 to 4.5 ml of follicular fluid and a mature oocyte and elevated serum estrogen (400 to 800 pg/ml) on the day after hCG treatment. At the time of follicular fluid recovery, there was more inhibin activity (204 +/- 14 U/10 ml) in the hMG/hCG-stimulated follicles than in unstimulated follicles of a control group of women (65 +/- 14, mean +/- standard error, P less than 0.05). Follicular fluid from follicles of hMG/hCG-treated women had lower estrogen and progesterone levels, compared with normal preovulatory women. These data show that treatment of patients with hMG/hCG is associated with elevated follicular fluid inhibin activity.

Adult↗

DNA polymerase and simian virus 40 infection of resting monkey cells: induction of aphidicolin resistant alpha-polymerase.

Tightly confluent monkey cell lines BSC-1 and CV-1 held in stale medium for several days exhibited an extremely low level of thymidine incorporation into cellular DNA. Yet, these cells contained a level of alpha-polymerase equal to about 15% of the level in rapidly dividing cells, and they still were capable of supporting replication of SV40 DNA. SV40 infection and culture in stale medium resulted in a four-fold induction of alpha-polymerase in CV-1 cells, whereas no change in alpha-polymerase level was observed in BSC-1 cells. Characterization of alpha-polymerase partially purified from infected CV-1 cells revealed that 80-90% of the enzyme activity was aphidicolin resistant. SV40 DNA replication in resting CV-1 cells, however, was aphidicolin sensitive. SV40 infection of resting CV-1 cells may induce an aphidicolin-resistant enzyme or lead to a modified alpha-polymerase species.

Animals↗

The occurrence of three different proteoglycan species in chick embryo cartilage. Isolation and characterization of a second proteoglycan (PG-Lb) and its precursor form.

Three different molecular species of proteoglycan (designated PG-H, PG-Lb, and PG-Lt) have been isolated from chick embryo epiphyseal cartilage. PG-H is a major proteoglycan of the tissue and identical, or nearly identical, with so-called cartilage-characteristic proteoglycan previously described in mammalian and avian cartilages. The third proteoglycan, PG-Lt, differs from the other two in containing disulfide-bonded collagenous polypeptides (Noro, A., Kimata, K., Oike, Y., Shinomura, T., Maeda, N., Yano, S., Takahashi, N., and Suzuki, S. (1983) J. Biol. Chem. 258, 9323-9331). The second proteoglycan, PG-Lb, consists of a core protein with Mr congruent to 52,000 dermatan sulfate copolymer chains with glucuronic acid/iduronic acid residues. Upon chondroitinase ABC digestion, the proteoglycan yields a protein-enriched core fraction of Mr congruent to 43,000. Its amino acid composition, tryptic peptide profile, and immunochemical properties indicate that PG-Lb is distinctly different from PG-H and PG-Lt in core protein structure. PG-Lb shows no specific binding with hyaluronic acid. Pulse-chase experiments with [3H]serine indicate that PG-Lb is first synthesized as a precursor form (pro-PG-Lb) that can be distinguished from PG-Lb by the production of a core molecule of Mr congruent to 52,000 after chondroitinase ABC digestion. This core molecule is labeled when [2-3H]mannose is used as a precursor, suggesting that it contains a glycoprotein type oligosaccharide. Since the core molecule from pro-PG-Lb is significantly larger in molecular weight than that from PG-Lb, the conversion of pro-PG-Lb to PG-Lb should involve scission of the polypeptide or possibly removal of mannose-containing oligosaccharide chain.

Amino Acids↗

Purification and characterization of adipose tissue S-100b protein.

We have purified S-100 protein from bovine brain using Ca2+-dependent affinity chromatography on N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7)-Sepharose (Endo, T., Tanaka, T., Isobe, T., Kasai, H., Okuyama, T., and Hidaka, H. (1981) J. Biol. Chem. 256, 12485-12489). By essentially the same procedure, W-7-Sepharose binding protein has been purified to apparent homogeneity from bovine abdominal adipose tissue. Electrophoretically, the purified protein from adipose tissue co-migrated with brain S-100b protein both in the presence and absence of sodium dodecyl sulfate and the protein was indistinguishable from brain S-100b region in terms of amino acid composition, two-dimensional tryptic peptide mapping and reactivity with anti-brain S-100b serum. Immunohistochemical analysis confirmed the existence of S-100b protein in the adipose cell where the protein seems to be located in both the nucleus and cytoplasm. Thus, the results indicate that the adipose cells contain the protein possibly identical with brain S-100b protein. In addition, the contents of S-100b protein in various rat tissues were measured by enzyme immunoassay method using the anti-bovine brain S-100b serum. Significant amounts of S-100b protein were found not only in the adipose tissue but also in the peripheral tissue such as trachea and skin. These observations suggest that S-100b protein should no longer be considered as a protein specific to nervous tissues.

Adipose Tissue↗

Transport of ions in erythrocytes infected by plasmodia.

Throughout its erythrocytic cycle the plasmodial parasite modifies the plasma membrane of its host cell. Some changes derive from parasite metabolism. Intraerythrocytic forms use glucose at more than 10-fold normal red cell rates. The H+ accompanying the lactate end-product is exported into the host cell cytoplasm by an electrogenic proton pump in the parasite membrane. This maintains a pH greater than 7.0 in the parasite cytoplasm, but lowers erythrocyte cytoplasmic pH from approximately 7.2 to 6.5. Ca2+ transport across parasite membranes is coupled to the proton pump, possibly a Ca2+/H+ antiporter. The Ca2+, Mg2+-ATPase and Na+,K+-ATPase activities of erythrocyte membranes from schizont-infected erythrocytes have been studied. Under optimal assay conditions (pH = 7.0; [ATP] = 1 mM; +/- calmodulin) membranes from infected cells showed a 30% reduction in Ca2+,Mg2+-ATPase activity but no difference from normal in Na+,K+-ATPase activity. The calmodulin levels of infected cells were depressed by about 30%. The [ATP] in the cytoplasm of infected erythrocytes was only 0.2 mM (as against 1.3 mM in normals) and at this ATP concentration the activities of both ATPases are only 30% of normal. Shifting the pH from 7.0 to 6.5 decreases Na+,K+-ATPase activity by an additional 50% but is without effect on the Ca2+,Mg2+-ATPase. The results provide a partial explanation for the increased Ca2+ permeability and altered Na+/K+ content of plasmodia-infected erythrocytes.

Animals↗

Detection of DNA replicase activity in rat thymoma cells.

A DNA polymerase which was active with unprimed poly(dC) as a template was detected in the DNA polymerase alpha fraction from rat thymoma tissue. The activity was dependent on rGTP and associated with ribonucleotide polymerizing activity. The activity was partially sensitive to aphidicolin and dideoxy GTP and resistant to alpha-amanitin. Thus, this is a rat "replicase" which is very similar to that found in mouse cells. The replicase activity with poly(dT) or phage fd single stranded circular DNA was greatly enhanced by the mouse replicase-stimulating factor beyond the species difference.

Animals↗

Staining of Plasmodium yoelii-infected mouse erythrocytes with the fluorescent dye rhodamine 123.

The cationic permeant fluorescent dye rhodamine 123 (R123) was used to stain Plasmodium yoelii-infected mouse erythrocytes. Fluorescence microscopic observations demonstrated that the parasite, but not the matrix of the infected erythrocyte, accumulated the dye. Differences in fluorescence intensity could not be found at the various developmental stages of the parasite; however, quantitation of the cell-associated dye revealed an increase in R123 uptake with parasite development. The retention of the parasite-associated dye, as measured by fluorescence microscopy and spectrophotometry after extraction of R123 with butanol, was markedly reduced by treatment of the infected erythrocytes with a proton ionophore, carbonyl cyanide m-chlorophenylhydrazone (CCCP), and an inhibitor of proton ATPase, dicyclohexylcarbodiimide (DCCD). These results indicate that the accumulation and retention of R123 in P. yoelii reflect the parasite membrane potential and suggest that the parasite plasma membrane has a membrane potential-generating proton pump.

Animals↗

Levels of inhibin-F activity and steroids in human follicular fluid from normal women and women with polycystic ovarian disease.

To examine inhibin-F activity (FSH-suppressing activity) in human follicular fluid of polycystic ovary (PCO) patients, 13 follicles from 5 documented PCO patients and an additional 31 follicles from normal women in the follicular phase of the menstrual cycle were sampled, and inhibin-F activity was measured in rat anterior pituitary cell cultures. Inhibin-F activity was measured in follicular fluid after stripping steroids from the fluids using treatment with dextran and activated charcoal. Estrogen, progesterone, and delta 4-androstenedione in the follicular fluid were also determined by RIA. Estrogen and progesterone levels in follicular fluid from PCO follicles 3.9 +/- 0.34 mm in diameter were comparable with those in follicular fluid obtained from viable follicles (which had a delta 4-androstenedione to estrogen ratio of 10 or less) from normal women. delta 4-Androstenedione levels in PCO follicles were higher (P less than 0.01) than those in viable and atretic follicles of normal women. Inhibin-F levels in PCO follicles were comparable to those in viable follicles, but significantly greater (P less than 0.01) than levels in atretic follicles of normal women. If inhibin-F levels in both atretic and viable follicles of normal women were pooled, the levels were less (P less than 0.05) compared to the level in PCO follicular fluid. As an additional control, follicular fluid was collected from 90 follicles of normal women throughout the menstrual cycle, and follicular size was determined as well as inhibin-F and steroid content. Small follicles less than 8 mm; (comparable in size to the PCO follicles examined) obtained at each stage represented 79%, 24%, 0%, and 94% of the total follicles obtained in the early to midfollicular, late follicular, preovulatory, and luteal phases of the cycle, respectively. Inhibin-F activity in the fluids of these follicles was less than that in PCO follicular fluid. The average inhibin content of all of the small normal follicles was 197 +/- 34 U/10 microliter, which was significantly less (P less than 0.05) than the level in PCO follicles (332 +/- 13 U/10 microliters). These data represent the first observation of inhibin-F activity in PCO follicular fluid and suggest the possibility of involvement of inhibin-F in bringing about low or normal basal levels of FSH in the presence of elevated basal LH levels often observed in PCO patients.

Animals↗

Influence of carbidopa, an aromatic amino acid decarboxylase inhibitor, on the development of autoimmune hemolytic anemia in NZB mice.

Causal relationships of carbidopa and its related drugs on the development of spontaneous autoimmune hemolytic anemia (AIHA) in NZB mice were studied, and the following results were obtained: 1) Long term treatment with carbidopa (3 mg/kg/day) and levodopa (30 mg/kg/day) neither accelerated nor suppressed the development of spontaneous AIHA in NZB mice. 2) In mice treated with carbidopa/levodopa (3/30 mg/kg/day), microhematocrit levels were lower than those in the control mice on and after 20 weeks of age and showed a significant decrease at 66 weeks of age (P less than 0.05). The average anti-RBC antibody titers reached the maximum level 8 weeks earlier than the control group. 3) Microhematocrit levels in the alpha-methyldopa (60 mg/kg/day)-treated group were higher than those in the control group, and at 66 weeks of age, they were decreased below that in the control group. The elevation of anti-RBC antibody titers was slower than that in the control group. As the reason for the weak effectiveness of alpha-methyldopa on the incidence of AIHA, it might be considered that the dosage employed was not sufficiently high enough and/or it may be due to the species difference between man and animals. Further studies are necessary in order to draw a conclusion on the AIHA-inducing ability of carbidopa.

Anemia, Hemolytic, Autoimmune↗

Induction and enhancement of endocytosis in HeLa cells by use of liposomes and Sendai virus envelopes.

Phosphatidylserine liposomes were interacted with cultured HeLa cells to investigate the ingestion of the liposomes into the cells. It was shown by electron microscopy that great enhancement in entrappment of the liposomes into the cells was induced by adding reassembled Sendai virus envelopes. In the presence of the reassembled virus envelopes the ratio of liposome-endocytic cells was over the half of the examined cells in thin sections. To differentiate the entrapped vesicles from cell-originated intracytoplasmic vesicles, uninfective virus particle was introduced as a mark and it was shown that the uninfective virus particle was entrapped with the liposomes into the cells. These results may suggest that reconstituted Sendai virus envelopes could change HeLa cell membranes to endocytic ones.

Animals↗

Incidence of hepatitis A virus infection among Japanese volunteers staying in developing countries.

Incidence of hepatitis A virus infection was studied serologically in 260 adult males and females sent for volunteer service to developing countries from Japan. Among 225 initially seronegative subjects, 29 (12.9%) were seropositive to hepatitis A virus at their return from two year's overseas service. Seroconversion occurred in none of 39 subjects who received two doses or more of prophylactic administration of human immunoglobulin. There were clinical records of acute non-B hepatitis in 10 subjects among 29 who seroconverted to hepatitis A virus, while in 6 among 231 who did not seroconvert.

Adult↗

A mapping technique for probing the structure of proteoglycan core molecules.

Our previous work showed that treatment of chick embryo cartilage proteoglycan (PG-H) with chondroitinase-AC II and keratanase yielded a protein-rich core fraction having enzymatically modified linkage oligosaccharides. The core sample has now been analyzed by tryptic peptide mapping, in which the isolated core sample contained in a single Coomassie blue-staining band from a dried slab gel is radioiodinated and treated with trypsin, and the resultant tryptic peptides are displayed two-dimensionally on a silica gel thin layer plate. The map thus obtained exhibited 22 major peptide spots, the resolution and location of which were reproducible. In order to identify regions of the core polypeptide from which the tryptic peptides are derived, PG-H was cleaved with clostripain under conditions that yield a hyaluronic acid-binding fragment with an apparent Mr = 150,000 and chondroitin sulfate-peptide clusters of smaller molecular sizes. Although the peptide maps of the two size classes of clostripain fragments differed significantly from each other, the patterns of spots, as a whole, were extensively similar to those observed with the intact core molecule. These results have provided additional evidence that PG-H has a single, nonvariable core protein structure. In addition, the technique used here will provide a versatile method for the identification of genetic types in this increasingly complex family of matrix macromolecules.

Animals↗

[Studies on the regulatory mechanism of ovarian hCG receptors and placental luteotropin in rats (author's transl)].

The influences of exogenous hCG on rat (pseudopregnant and pregnant) ovarian hCG receptors and the presence of hCG-like material in the rat placenta were investigated. Ovarian hCG receptors in immature rats induced by PMS-hCG priming maintained a binding activity for about 20 days after hCG injection. The administration of hCG to pseudopregnant rats caused time and dose related changes in ovarian 125I-hCG binding percentage and the concentration of serum progesterone. The injection of hCG produced a slight decrease in the binding percentage and was followed by a marked decrease. The measurement of ovarian tissue hCG concentration by radioimmunoassay suggested that the initial loss of binding percentage was associated with the occupancy of receptors by the exogenous hCG, but the major loss of binding percentage might not be attributed to the receptor occupancy. The active process might be involved in the mechanism of receptor loss. The progesterone concentration in sera had a tendency to increase after the injection of 25IU hCG, while it decreased after the injection of 125IU hCG. The 125I-hCG binding percentage to ovaries and the serum concentration of progesterone reached their maximal values from mid to late pregnancy and declined as parturition approached. After the injection of hCG into pregnant rats, the binding percentage rapidly decreased and the serum concentration of progesterone increased. Thereafter the binding percentage recovered on the 4th day after the injection of hCG. On that day, though the concentration of hCG in ovarian tissue was barely detectable, the binding percentage was significantly low compared to the control level. The rat placenta (8th to 21st day of pregnancy) contained hCG-like activities, but they could not be detected in peripheral blood. In the treatment of hCG-antibody in pregnant rats, the binding percentage did not change but the serum concentration of progesterone showed a tendency to decrease to the control level. These studies show that hCG receptors exist as free sites in pregnant and pseudopregnant rat ovaries and may be regulated by down regulation after the treatment of large amounts of exogenous hCG. We found that hCG-like material in rat placenta determined by radioimmunoassay might serve as a luteotropin during pregnancy.

Animals↗