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Biomedical subjects

H Kagawa

Publications and source records attributed to H Kagawa.

At least 127 records · Page 7Linked to original sources

Classification and Structural Comparison of Full-Length cDNAs for Pathogenesis-Related Proteins.

Fourteen cDNA clones of pathogenesis-related (PR) proteins, PR1a and PR1b of tobacco were obtained and classified into six groups based on restriction enzyme maps. To assign the groups to different classes of PR1 proteins, all the clones were partially sequenced and compared with amino acid sequences of PR1a and PR1b. Two groups of these corresponded to PR1a and four to PR1b. The results indicate that there are at least two kinds of PR1a mRNAs and four kinds of PR1b mRNAs. In fact, one cDNA insert hybridized to at least six to seven DNA fragments in restriction enzyme fragments of Samsun NN genomic DNA, indicating that the PR1 protein genes exist as a multigene family in the tobacco genome. Two sequences of essentially full-length cDNAs for PR1a and PR1b were determined and compared. The coding sequences of two cDNAs share 93% homology and the deduced amino acid sequences of PR1a and PR1b precursors, which are synthesized as larger precursors containing signal peptides, are 91% homologous. The homology of mature PR1a and PR1b regions is higher than that of larger precursors, 94% in the nucleotide sequence and 93% in the amino acid sequence, whereas that of the signal peptide regions is 80 and 90%, respectively. The hydropathy patterns and the secondary structures predicted by Chou-Fasman rules are similar to tomato PR protein in the half-side of the C terminus, which suggests that the half-C terminus side is important for the function of PR1 proteins.

Journal Article↗

Immunocytochemistry on the localization of 5-hydroxytryptamine in monkey and rabbit taste buds.

Immuno-electron microscopy with the protein A-gold method demonstrated immunoreactive gold particles against 5-hydroxytryptamine localized in cored vesicles aggregating around presynaptic terminals of the gustatory cells in monkey and rabbit taste buds. The positive reactions were also found in the intragemmal and subepithelial nerve fibers. The role of these cored vesicles in taste transduction is still uncertain but the data strongly suggest that they may participate in a serotonergic modulation of a cholinergic synaptic transmission from the gustatory cells to the nerve endings.

Animals↗

Effects of human recombinant interleukin 2 in patients with chronic hepatitis B: a preliminary report.

Five patients with HBe antigen-positive chronic active hepatitis were treated with recombinant interleukin 2 (r-IL2) for 3 to 4 wk. r-IL2 inhibited HBV replication during therapy as judged by decrease of serum DNA polymerase activity. Two of five patients lost HBeAg from serum. Furthermore, characteristic increases of serum aminotransferase were observed during r-IL2 therapy. This rise might have been mediated by enhanced immune responses to hepatitis B virus and increased rate of lysis of the infected hepatocytes. No consistent changes in immune markers such as OKT4 positive cells or OKT8 positive cells were demonstrated during or after the therapy. The activities of natural killer cells gradually increased during therapy, although they tended to decrease within 1 wk after the start of r-IL2 injections. In two responders who lost HBeAg from serum, the natural killer activity was high before the therapy, compared with nonresponders. Furthermore, the percentage of OKT4-positive cells in the responders was increased within 1 to 2 wk after starting r-IL2 injections. These findings suggest that the responders might be immunologically different from the nonresponders. Recombinant-IL2 therapy over short periods did not result in complete clearance of hepatitis B virus. Further studies with high doses of r-IL2 given over longer periods are warranted.

Adult↗

Immunoelectron microscope observations on secretion of human placental lactogen (hPL) in the human chorionic villi.

Immunoelectron microscope localization of human placental lactogen (hPL) was investigated in the chorionic villi from week 7 to full-term gestation with the protein A-gold technique. With specific antiserum against hPL, immuno-reactive gold particles were found to be preferentially located in Golgi-derived, electron-dense small granules of 80-180 nm in the syncytiotrophoblast. Our electron micrographs indicate that these small granules increase in number in the course of gestation and are released by exocytosis from the apical cell surface. The present study reveals that hPL is segregated from the Golgi apparatus, stored in the syncytiotrophoblast as secretory granules, and released into the maternal blood.

Chorionic Villi↗

Rabbit autoantibodies to actin induced by immunization with modified homologous actins.

Actin is a major antigen involved in the reaction of smooth muscle antibody positive sera from patients with chronic active hepatitis. In the present study, actin extracted from rabbit skeletal muscle was denatured by sodium dodecyl sulfate and was immunized into the rabbit, a homologous animal for actin. The rabbits, thus immunized, produced antibodies reactive with actins of homologous and heterologous animals. In addition, the antibodies showed reactivity with autologous actin. It indicates that the denatured homologous actin is capable of terminating immunological tolerance to actin and induces formation of autoantibody to rabbit actin. This phenomenon may be implicated in the occurrence of anti-actin antibody in sera from patients with chronic liver disease and several other diseases.

Actins↗

[Intra-tumoral injection therapy in patients with hepatocellular carcinoma].

Ten patients with hepatocellular carcinoma (HCC) received intra-tumoral injection of OK-432 (6 patients), 99.5% ethanol (2 patients) or both (2 patients). Under ultrasonographic control, a PTC needle (22 G) was inserted percutaneously into the tumor and OK-432, which was prepared with a solution of Su-strain Streptococcus pyogenes A3, or 99.5% ethanol was injected. Patients were injected with OK-432 repeatedly at one-to two-week intervals (up to 5 times) for a total duration of 5 to 15 weeks. The degree of skin test reaction for Streptococcus pyogenes was increased in all patients after the treatment. Over 40% tumor regression was noted in 6 out of 9 patients who received intra-tumoral injection of OK-432. Complete regression was noted in one patient. Before treatment, Interleukin-2 (IL-2) and lymphokine-activated killer (LAK) cell activity in peripheral blood lymphocytes decreased in HCC patients. Two of 6 patients showed markedly increased activity of LAK-cells one week after treatment with OK-432. One other patient had moderately increased LAK-cell activity after treatment with OK-432. No increase in LAK-cell activity was seen in 3 patients who received intra-tumoral injection of ethanol. An especially increased response of LAK-cell activity was seen in patients with small-sized HCC (diameter below 5 cm).

Administration, Topical↗

Immunocytochemical localization of factor VIII-related antigen in the human umbilical vein.

The immunocytochemical localization of factor VIII-related antigen in the human umbilical vein was investigated with the light microscopic immunoperoxidase and immunoelectron microscopical protein A-gold techniques. The light microscopic observation showed that peroxidase reactions were found exclusively in the endothelia of the vessel. By the protein A-gold method, immunoreactive gold particles were located in endothelial specific granules (Weibel-Palade bodies). These data suggest that endothelial specific granules are storage sites of factor VIII-related antigen.

Antigens↗

Ultrastructural and histochemical observations regarding the ovarian follicles of the amago salmon (Oncorhynchus rhodurus).

Pre- and postovulatory ovarian follicles of the amago salmon were investigated with special consideration given to the steroid production site. In the preovulatory specimens, the special thecal cells contained abundant mitochondria and smooth endoplasmic reticulum, as in the case of other steroid-producing cells, while the granulosa cells developed rough endoplasmic reticulum like protein-secreting cells. Remarkably ultrastructural changes occurred in the granulosa and the special thecal cells during the oocyte maturation stage; dilation of rough endoplasmic reticulum in the former cell and further development of smooth endoplasmic reticulum in the latter one. Histochemical reactions of delta 5-3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity were only positive in the special thecal cells of the postovulatory follicles. These results strongly suggest that the special thecal cells are the major sites of estrogen precursor synthesis in the amago salmon ovary.

3-Hydroxysteroid Dehydrogenases↗

Immunoelectron microscopic localization of growth hormone in the pituitary glands of two teleosts, tilapia (Sarotherodon mossambicus) and amago salmon (Oncorhynchus rhodurus).

Growth hormone (GH) cells were investigated with the protein A-gold technique on the pituitary glands of tilapia (Sarotherodon mossambicus) and amago salmon (Oncorhynchus rhodurus). By the use of specific antiserum against tilapia GH to both species, the immunoreactive gold particles were demonstrated to be preferentially located on the secretory granules of the GH cells. Specimens fixed only with periodate-lysine-paraformaldehyde (PLP) preserved the hormonal antigenicity well. Osmium postfixation, although considerably reducing the antigenicity and thus resulting in a decrease in number of the gold particles on the GH cells, gave much more satisfactory ultrastructural preservation and immunoreactive localization of immunoreactive material. This investigation demonstrated that, after combined fixation with PLP and PLP-osmium, we could determine the function of a given cell type in various endocrine organs as well as the precise antigenic sites in such cells.

Animals↗

Synthesis of flagellin and hook subunit protein in flagellar mutants of Escherichia coli K12.

We have examined Escherichia coli K12 flagellar mutants affected in each of 29 different loci for the synthesis of flagellin and hook subunit protein. Immune precipitation experiments were employed by treating cell extracts with antiserum against each protein. Flagellin was synthesized in mutants defective in genes flaS , flaT, flaU and flbC . The flaE and flaZ mutants produced small amounts of flagellin, while all the other mutants failed to produce any detectable amount of flagellin. Hook subunit protein was found in most mutants including those defective in genes flaA , flaB , flaC, flaD , flaE , flaG , flaH , flaL , flaM , flaN , flaO , flaP, flaQ , flaR , flaS , flaT, flaU , flaV , flaW , flaX, flaY , flaZ , flbA , flbC , flbD , and hag but not in mutants of flaK , flaI , and flbB . The results conform to the predictions made by our previous indirect gene fusion study ( Komeda 1982).

Bacterial Proteins↗

In vitro estradiol-17 beta and testosterone production by ovarian follicles of the goldfish, Carassius auratus.

In vitro production of estradiol-17 beta and testosterone by ovarian follicles of goldfish (Carassius auratus) at different developmental stages in response to human chorionic gonadotropin (HCG) was examined using 18-hr incubations. The vitellogenic follicles (primary and secondary yolk stage) produced estradiol-17 beta in response to HCG; follicles at the secondary yolk stage produced about five times more estradiol-17 beta than the primary yolk stage follicles. However, tertiary yolk stage follicles, which could be induced to mature in vitro by HCG treatment, did not produce estradiol-17 beta. HCG also stimulated testosterone production by follicles at all stages of development; maximal production was observed in tertiary yolk stage follicles. An indirect assessment of aromatase activity at each stage was made. Vitellogenic follicles (primary and secondary yolk stage) produced estradiol-17 beta when incubated with testosterone, the larger follicles producing about three times more estradiol-17 beta. Only low levels of estradiol-17 beta were produced by tertiary yolk stage follicles. Addition of HCG to primary yolk stage follicles slightly but significantly enhanced estradiol-17 beta production; however, no enhancement was seen with secondary and tertiary stage follicles. These results suggest that the decrease in estradiol-17 beta production in tertiary yolk stage follicles may be partly due to a decrease of aromatase activity at this stage.

Animals↗

Bacteriophage chi sensitivity and motility of Escherichia coli K-12 and Salmonella typhimurium Fla- mutants possessing the hook structure.

The production of hook protein and flagellin in 29 Fla- mutants of Escherichia coli K-12 was determined by the complement fixation assay. Six mutants produced hook protein, and four of them also produced flagellin. A flaE mutation was introduced into these fla mutants carrying the hook structure. All of these mutants made polyhooks and were used as hosts for a newly isolated host-range mutant of chi phage that has a high affinity for the hook structure. All except one mutant produced significant amounts of progeny phages. A flaD flaE double mutant was that exception which did not yield significant amounts of progeny by the phage propagation method. All of the flaE double mutants produced comparable amounts of polyhooks, and no qualitative difference was detected between chi-sensitive and chi-insensitive mutants by the complement fixation assay. Accordingly, it was thought that the polyhook of the flaD flaE mutant had a mechanical defect for chi phage infection. This assumption was confirmed by tethered-cell experiments; the flaD flaE mutant did not rotate. These results are well explained by a proposed regulation pathway of flagellar genes. flaE mutants can express other genes which govern the final step of the flagellar morphogenesis, whereas flaD mutants cannot rotate, possibly because the mocha operon is not expressed. The results obtained in E. coli were also found to be applicable to Salmonella typhimurium.

Bacterial Proteins↗

[Demonstration of specific gonadotropin binding sites of chum salmon (Oncorhynchus keta) in membrane preparations of granulosa cells from the ovary of amago salmon (Oncorhynchus rhodurus)].

During late vitellogenesis, radioactive iodine labelled Salmon gonadotropin binds to theca--or granulosa--membrane preparations from amago Salmon ovary. In granulosa, native hormone inhibits iodinated GTH binding, thus indicating a specific binding (70% of total binding). In theca, a lower specific binding is found while non specific binding is higher: contamination by granulosa cells, on one hand, the possible heterogeneity of theca, on the other, could account for these results.

Animals↗

Isolation and characterization of nucleic-acid-like polymers containing glucose from Ehrlich ascites tumor cells.

1. Novel nucleic-acid-like polymers containing glucose, glucose 'nucleic acid', were isolated from cytoplasmic particles of Ehrlich ascites tumor cells. It is assumed that the basic structure of the polymers is similar to those of known nucleic acids except that their sugar constituents are glucose derivatives instead of ribose or deoxyribose. 2. When the glucose 'nucleic acid' was incubated with venom phosphodiesterase, four kinds of glucose 'nucleotides' containing bases of cytosine, adenine, guanine and thymine were obtained. 3. When the polymers were incubated with both venom phosphodiesterase and alkaline phosphatase, inorganic phosphates of glucose 'nucleotides' were removed and each nucleotide was converted to a glucose 'nucleoside'.

Animals↗

Plasma 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one levels during sexual maturation of amago salmon (Oncorhynchus rhodurus): correlation with plasma gonadotropin and in vitro production by ovarian follicles.

Plasma gonadotropin (GtH) and 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one (17 alpha, 20 beta-diOH-prog) levels were assessed by radioimmunoassay during the sexual maturation of female amago salmon (Oncorhynchus rhodurus). Both GtH and 17 alpha, 20 beta-diOHprog levels were low in vitellogenic females (June to September) and in those with full-grown immature oocytes and were elevated in mature and ovulated females to 40 ng/ml for GtH and 50-70 ng/ml for 17 alpha, 20 beta-diOHprog. In vitro production of 17 alpha, 20 beta-diOHprog by ovarian follicles and its stimulation by partially purified chinook salmon gonadotropin (SG-G100) was examined monthly using 18 hr incubations. In June and July, levels were too low to detect (less than 30 pg/ml) in media from all treatment groups. In August and September, SG-G100 at a concentration of 1 microgram/ml stimulated low levels of production (0.2-0.3 ng/ml). Full-grown, immature follicles in October showed a dose-response production of 17 alpha, 20 beta-diOHprog, averaging over 10 ng/ml when incubated with 1 microgram/ml SG-G100. Postovulatory follicles (1-2 days after ovulation) produced large amounts of 17 alpha, 20 beta-diOHprog, averaging over 100 ng/ml with 1 microgram/ml SG-G100. These results are discussed in relation to previous work on amago salmon and other species and together indicate that 17 alpha, 20 beta-diOHprog is the major steroid responsible for oocyte maturation in amago salmon, produced as the follicular mediator of gonadotropin.

Animals↗

Relationship between seasonal plasma estradiol-17 beta and testosterone levels and in vitro production by ovarian follicles of amago salmon (Oncorhynchus rhodurus).

Plasma estradiol-17 beta and testosterone levels were assessed by radioimmunoassay during the sexual maturation of female amago salmon (Oncorhynchus rhodurus). Estradiol-17 beta levels gradually increased during vitellogenesis (June to September), reached a peak in September (about 16 ng/ml) and rapidly decreased in mature and ovulated fish (about 3-4 ng/ml) in October. The seasonal pattern of plasma testosterone levels lagged behind and followed that of estradiol-17 beta during vitellogenesis, but levels remained high in mature and ovulated fish (90-110 ng/ml). Estradiol-17 beta levels and the gonadosomatic index (GSI) values correlated well during vitellogenesis: GSI values showed a linear increase, and reached a peak (29.9 +/- 1.4) in October. Values were extremely low in ovulated fish (1.2 +/- 0.2). In vitro production of estradiol-17 beta and testosterone by ovarian follicles in response to partially purified chinook salmon gonadotropin (SG-G100) was examined monthly using 18-h incubations. Throughout the vitellogenic period SG-G100 stimulated both estradiol-17 beta and testosterone production: the steroidogenic response of follicles increased from June (about 2 ng/ml estradiol-17 beta; 0.1 ng/ml testosterone) to September (about 10 and 14 ng/ml, respectively). In October full-grown immature follicles which could be induced to mature in vitro by hormone treatment produced large amounts of testosterone (about 130 ng/ml) but not estradiol-17 beta. Postovulatory follicles also produced testosterone but the values were low (10 ng/ml) compared with full-grown immature follicles. Very low levels of estradiol-17 beta were produced by postovulatory follicles.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗