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Biomedical subjects

H Mizuma

Publications and source records attributed to H Mizuma.

48 records · Page 3Linked to original sources

Thyroid hormone affects the hydrolysis of inositol phospholipids in the rat hypothalamus.

We have attempted to elucidate the effect of thyroid hormone on phospholipase C-linked inositol phospholipid hydrolysis in the rat hypothalamus. Hypothalamic slices of each animal, euthyroid control, hypothyroid, and thyroxine (T4)-supplemented hypothyroid rats were labeled with [3H]myoinositol in the presence of 5 mM LiCl, and then incubated for 60 min in KHG buffer containing either vehicle or 1 mM ouabain, a Na-K ATPase inhibitor. Hypothyroidism caused a significant increase in both basal and ouabain-stimulated accumulation of [3H]inositol phosphate ([3H]IP) in hypothalamic slices, whereas supplement with T4 to hypothyroid rats resulted in a complete restoration of hypothalamic [3H]IP formation to the value of euthyroid control. The present results indicate that thyroid hormone affects phospholipase C-linked inositol phospholipid hydrolysis in the hypothalamus, suggesting that negative feedback action of thyroid hormone may occur at a post-receptor site in the hypothalamus.

Animals↗

Inhibitory effects of okadaic acid on thyrotropin and prolactin secretion from rat anterior pituitaries.

The present study was undertaken to elucidate the effects of okadaic acid, a potent inhibitor of protein phosphatases, on thyrotropin (TSH) and prolactin (PRL) secretion, and on the hydrolysis of inositol phospholipids in rat anterior pituitaries. Preincubation of anterior pituitaries with okadaic acid caused a dose dependent decrease in TRH- and K(+)-induced TSH secretion, whereas basal secretion of TSH was not affected by pretreatment with okadaic acid. In contrast, okadaic acid resulted in a marked inhibition in both basal, and TRH- and K(+)-stimulated PRL release from anterior pituitaries. In addition, pretreatment with okadaic acid caused a slight, but significant decrease in the formation of [3H]inositol phosphate ([3H]IP) in rat anterior pituitaries. The present study suggests that okadaic acid blocks the release of TSH and PRL by inhibiting Ca2+ influx and that inhibitory effects of okadaic acid on PRL release are, at least in part, due to the inhibition of inositol phospholipid hydrolysis.

Animals↗

Identification of inositol 1,4,5-trisphosphate binding in human pheochromocytoma.

An attempt was made to identify a specific binding of inositol 1,4,5-trisphosphate (IP3) to human chromaffin cell tumors. Crude microsomal fractions of pheochromocytomas possessed significant binding sites for [3H]IP3 with high- and low-affinity constants (Kd = 5.46 and 538 nM, respectively). The binding to these sites was rapid, saturable (reached equilibrium by 1 min at 0 degrees C) and reversible. Competition studies with other inositol phosphate analogs indicated the stereotypic specific binding for IP3. Although no significant difference was observed in the binding densities of IP3 between pheochromocytomas and adrenal nonfunctioning adenomas, the values of high-affinity constants were significantly lower in the former than the latter group (5.77 +/- 1.07, n = 5 vs. 11.30 +/- 1.98 nM, n = 4, p less than 0.05). The present data indicate that characteristics of IP3 binding sites are congruous with their receptor functions and that changes in its binding sites may contribute to the biochemical dissimilitudes in pheochromocytomas.

Adenoma↗

Hypothyroidism inhibits the formation of inositol phosphate in response to carbachol in the striatum of adult rat.

The effects of hypothyroidism on the muscarinic cholinergic receptor-coupled inositol phospholipid hydrolysis in the adult rat brain were examined. Tissue slices of striatum, hippocampus, and cortex from either euthyroid or hypothyroid rats were labeled with [3H]myoinositol and incubated with carbachol, a muscarinic cholinergic agonist. In other experiments, crude plasma membranes of each brain region obtained from either euthyroid or hypothyroid rats were incubated with [3H]N-methylquinuclidinyl benzilate ([3H]NMeQNB), a muscarinic cholinergic antagonist, in the presence or absence of atropine. Carbachol produced a significant increase in [3H]inositol phosphate ([3H]IP) formation in each brain region in a dose dependent manner. Hypothyroidism caused a marked decrease in carbachol-stimulated [3H]IP formation in the striatum, whereas it did not affect the formation of [3H]IP in the cortex or hippocampus. In contrast, the affinity constant and the maximal binding of [3H]NMeQNB to plasma membranes in these regions were not changed by hypothyroidism. The present results suggest that thyroid hormones might participate in regulating the muscarinic cholinergic neurotransmission in the striatum of adult rat.

Animals↗

B-cell activation in HIV infection: relationship of spontaneous immunoglobulin secretion to various immunological parameters.

Peripheral blood mononuclear cells from HIV-infected individuals spontaneously secrete elevated levels of IgG, IgM and IgD. This increased level of synthesis and secretion is similar in HIV-infected subjects with no or few symptoms, in ARC patients and in AIDS patients. Thus, abnormal B-cell activation is characteristic of patients with mild as well as severe manifestations of HIV infection. The level of spontaneous cellular secretion of IgG, IgM and IgD correlates with serum levels of these isotypes. Levels of spontaneous cellular secretion of IgG and IgM correlate negatively with the percentage but not with the absolute number of T4-positive cells and correlate positively with the percentage but not with the absolute number of T8-positive cells. The data suggest that the proportional distribution of these T-cell subsets is a critical factor in the B-cell dysregulation leading to overproduction of IgG and IgM. On the other hand, spontaneous IgD secretion correlates with neither the percent nor the absolute number of T4 or T8 cells suggesting that the increase of IgD-secretion by B cells is independent of the T-cell defects. The data imply that more than one mechanism underlies the B-cell activation in HIV-infected individuals.

AIDS-Related Complex↗

Potential use of serotherapy in the prevention and treatment of infection with the human immunodeficiency virus.

While prevention of infection with the human immunodeficiency virus (HIV) using prophylactic immunization and treatment with anti-viral drugs would appear to be the methods of choice for the prevention and treatment of this infection, neither safe and effective vaccines nor anti-viral agents have yet been developed. A third approach should thus be considered which could be employed both for prophylaxis and treatment of this disease. This approach utilizes specific, anti-HIV antibodies, passively administered, to prevent and/or slow the infectious process. The disadvantages of using xenogeneic antibodies and the advantages of using human antibodies are discussed. The need for large quantities of human antibodies to HIV necessitates the production of cell lines producing these antibodies. The various techniques of producing these lines are summarized. Finally, preliminary data supporting the feasibility of producing human cell lines producing antibody to HIV are presented.

Acquired Immunodeficiency Syndrome↗

Serum IgD elevation is an early marker of B cell activation during infection with the human immunodeficiency viruses.

Serum IgD levels in individuals infected with the human immunodeficiency viruses (HIV) were studied as a means of monitoring the character and timing of B cell activation in individuals with this infection. Significantly increased levels of IgD were characteristic of homosexual men who were HIV seropositive but asymptomatic or mildly symptomatic. The hyper IgD globulinaemia became progressively more pronounced in patients with increasingly severe infection and reached its most marked level in patients with AIDS-related complex (ARC). In ARC patients, IgD levels were increased 8.8-fold above normal which was disproportionately greater than the 2.4-fold increase in IgG, the 1.8-fold increase in IgA and the 1.6-fold increase in IgM. IgD levels declined in AIDS patients (although remained elevated compared to controls). The data suggest that an unusual type of B cell activation is responsible for the unique pattern of hypergammaglobulinaemia seen in this disease and that the B cell activation occurs early in the pathogenesis of HIV infection, often before development of symptoms, and continues throughout the course of infection.

AIDS-Related Complex↗

Elicited antibody nature of human monoclonal protein with anti-streptolysin O activity--analysis with monoclonal anti-idiotype antibody.

Sera from 7 patients with multiple myeloma having antistreptolysin O (ASO) activity in high titers were detected by a streptolysin O (SLO) inhibition assay. However, activity was in low titer when assayed by a passive agglutination assay. The discrepancy between these 2 assays raised some doubts as to whether these monoclonal proteins (M.protein) bond to SLO in the same manner as elicited antibodies. Immunochemical analysis and idiotope analysis using monoclonal antibody to one of these M.proteins strongly suggest that M.protein with ASO activity bind to SLO in a manner similar to elicited antibody. The discrepancy between the 2 assays might be due to differences in the antigenic structure of different forms of the SLO molecule.

Antibodies↗

A nondenaturing protein map of human plasma proteins correlated with a denaturing polypeptide map combining techniques of micro two-dimensional gel electrophoresis.

Human plasma proteins were separated by combining four types of two-dimensional electrophoresis (2-DE) techniques to obtain systematic information on proteins and their constituent polypeptides. A micro gel system was employed to facilitate the analysis. A plasma sample was first analyzed under nondenaturing conditions of electrophoresis (Type I 2-DE) to characterize the properties of proteins under physiological conditions. The sample was then analyzed, employing nondenaturing isoelectric focusing in the first dimension and sodium dodecyl sulfate (SDS) electrophoresis in the second dimension (Type II 2-DE), to study the dissociation of noncovalently bound protein subunits. In the third type of 2-DE (Type III 2-DE), proteins were separated by nondenaturing isoelectric focusing and treated with urea/mercaptoethanol/SDS and then subjected to second-dimension SDS electrophoresis, to study the dissociation of disulfide-bonded polypeptides. In the fourth type of 2-DE (Type IV 2-DE), the conditions of denaturing 2-DE were employed; the sample was treated with SDS-mercaptoethanol-urea-Nonidet P-40, separated by denaturing isoelectric focusing, and then subjected to SDS electrophoresis. The combined 2-DE technique will be useful to construct a comprehensive database of plasma proteins combining a "nondenaturing protein map" (a protein map) and a "denaturing protein map" (a polypeptide map).

Adult↗

Identification of cyclo(His-Pro)-like immunoreactivity in human follicular fluid: correlation with steroid and peptide hormones.

OBJECTIVE: The purpose of this study was to evaluate human follicular fluid (FF) for the presence of cyclo(His-Pro)-like immunoreactivity (CHP-LI). After verifying its presence, we quantitated the levels and investigated correlations with other follicular parameters, including hormone levels. METHODS: Follicular fluid was collected from female volunteers undergoing controlled ovarian hyperstimulation. Fluid was collected by follicular puncture, either transvaginally (in vitro fertilization) or laparoscopically (gamete intrafallopian transfer) at the time of oocyte retrieval (N = 137). Follicular size was determined ultrasonographically. Assays for steroid and peptide hormones were determined with commercially available radioimmunoassay kits. CHP-LI was measured using a previously reported assay; parallel dilution curves and column chromatography aided in immunoidentity. RESULTS: The mean FF CHP-LI concentration (13.10 +/- 1.83 nmol/L, N = 137) was greater than the corresponding serum values (9.42 +/- 2.45 nmol/L; N = 21) (P < .05). Large follicles (20 mm or greater; 14.45 +/- 1.74 nmol/L) contained significantly more CHP-LI than either medium follicles (16-19 mm; 11.51 +/- 1.88 nmol/L) or small follicles (15 mm or smaller; 10.83 +/- 2.12 nmol/L) (P < .05). Positive correlations were found between FF CHP-LI values and corresponding FF progesterone and prolactin concentrations (r = 0.67 and 0.62, respectively; P < .05). CONCLUSION: Mean CHP-LI levels in the FF are greater than those in the corresponding serum. We suggest that the neuropeptide may be originating from either peptidase cleavage of precursor peptides or from granulosa cell production.

Adult↗