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Biomedical subjects

M Huang

Publications and source records attributed to M Huang.

At least 325 records · Page 18Linked to original sources
PubMed1990

Characterization of the gag/fusion protein encoded by the defective Duplan retrovirus inducing murine acquired immunodeficiency syndrome.

Murine acquired immunodeficiency syndrome is induced by a defective retrovirus. Sequencing of this defective viral genome revealed a long open reading frame which encodes a putative gag/fusion protein, N-MA-p12-CA-NC-COOH, (D. C. Aziz, Z. Hanna, and P. Jolicoeur, Nature (London) 338:505-508, 1989). We raised a specific antibody to the unique p12 domain of this gag fusion precursor, Pr60gag. We found that Pr60gag was indeed encoded by the defective viral genome both in cell-free translation reticulocyte extracts and in infected mouse fibroblasts. Pr60gag was found to be myristylated, phosphorylated, and attached to the cell membrane, like other helper murine leukemia virus (MuLV) gag precursors. Pr60gag was not substantially cleaved within the nonproducer cells and was not released from these cells. However, in the presence of helper MuLV proteins, it formed phenotypically mixed particles. In these particles, Pr60gag was only partially cleaved. In helper MuLV-producing cells harboring the defective virus, a gag-related p40 intermediate was generated both intracellularly and extracellularly. In these cells, Pr60gag appeared to behave as a dominant negative mutant, interfering with proper cleavage of helper Pr65gag. Our data indicate that Pr60gag is a major (and possibly the only) gene product of the defective murine acquired immunodeficiency syndrome virus and is likely to harbor some determinants of pathogenicity of this virus.

Animals↗
PubMed1990

Functional elements of the steroid hormone-responsive promoter of mouse mammary tumor virus.

Transcription from the promoter of mouse mammary tumor virus is subject to induction by several classes of steroid hormones as well as to repression by a negative regulatory element present in the long terminal repeats of proviral DNA. In order to characterize the functional elements of the promoter that in some way must respond to these regulatory signals, a number of promoter mutations were constructed, including a set of linker-scanning mutations across the entire promoter region. Analysis of these mutated promoters with a transient-transfection assay defined at least three mutation-sensitive promoter elements that are required for both basal and hormone-induced transcription. One mutation-sensitive region contains a TATA element located at approximately position -30 with respect to the start of transcription. A second mutation-sensitive region contains two 10-base-pair direct repeats located between positions -60 and -38, within which are embedded three copies of octamer-related sequences; complete disruption of this region of the promoter leads to a more severe decrease in transcription than do any of the linker-scanning mutations, suggesting that the repeated sequences may be at least partially functionally redundant. Gel electrophoresis mobility shift assays were used to demonstrate specific binding of a nuclear protein to this region of the promoter. A third mutation-sensitive region contains a binding site for nuclear factor 1 (NF-1) located between positions -77 and -63. Site-directed mutations in the NF-1-binding site which increase the apparent affinity of NF-1 for the promoter in vitro do not decrease the hormone dependence of transcription, suggesting that transcriptional activation mediated by steroid hormone-receptor complexes cannot be explained by facilitation or stabilization of the interaction of promoter sequences with NF-1 and consistent with the idea that binding of NF-1 is not rate determining in transcription from the mouse mammary tumor virus promoter. None of the promoter mutations functionally separates basal from glucocorticoid-induced transcription, suggesting that hormone induction does not make the promoter independent of any of the DNA-binding factors required for its basal activity.

Animals↗
PubMed1990

Outer hair cells as potential targets of inflammatory mediators.

Inner ear sequelae with temporary or permanent sensorineural hearing loss can result from inflammatory processes in the middle ear. Loss of outer hair cells in the base of the cochlea has been noted in otitis media, but it is not known how this damage occurs. Evidence supports the permeability of the round window membrane to substances mediating inflammation in the middle ear, and the presence of white blood cells has been reported in the perilymph. In the present study, the potential cytotoxic effects of two representative inflammatory mediators, endotoxin and free radicals, have been evaluated by use of short-term culture of isolated outer hair cells from the guinea pig cochlea model. Incubation with endotoxins from two gram-negative pathogens increased the rate of hair cell death fourfold to sixfold. Free radicals (generated by exposure of cells to UV light or by excitation of intracellular fluorescent dyes) produced morphologic damage to hair cells within 60 seconds. These latter effects were delayed by addition of free-radical scavengers. It is concluded that inflammatory mediators are cytotoxic to hair cells and therefore are potentially ototoxic if permeating the round window membrane.

Animals↗
PubMed1990

[Effect of reserpine on both CL seizures and cAMP levels in seizure mice brain].

Reserpine injection (1 mg/kg i.p.) could not only significantly reduce cAMP levels in both cerebral cortex and hippocampus in naive WC1 mice, but also potentiate the degree of seizures elicited by CL. Reserpine injection could significantly shorten CL-seizures latency and decrease its threshold, too. Reserpine pretreatment could also diminish CL seizure-induced accumulation of cAMP. These results indicated the cAMP level elevated by seizure activity was associated with monoamine neurotransmitter activity, and the seizure-induced accumulation of cAMP might take part in the process which eliminated the spread of seizure discharges and speeded up the termination of seizures.

Animals↗
PubMed1990

[Variation of cAMP in cerebral cortex and hippocampus in seizure mice induced by coriaria lactone].

To study the relationship between cAMP and epilepsy, we investigated the effects of coriaria lactone (CL) on the levels of cAMP in cerebral cortex and hippocampus. Fifty normal WC1 mice were divided into 5 groups. All groups were injected with CL (2.5 mg/kg, i.m.), except the control group which was injected with NS. The concentrations of cAMP were measured after the animals had been immersed in liquid nitrogen to die at different phases of seizure development induced by CL (before seizures, in mild seizures, during severe seizures, and after severe seizures). The results indicated that the levels of cAMP in both cerebral cortex and hippocampus were significantly increased (1.73-fold and 1.33-fold) in severe seizure, and they continuously increased (1.85-fold and 1.45-fold) after severe seizures. On the other hand, no changes of cAMP were observed in mild seizures and before seizures. These data suggested that accumulation of cAMP is probably the result of seizure activity rather than the cause of seizure.

Animals↗
PubMed1990

Selectivity for binding of peptide analogs to vascular receptors for vasoactive intestinal peptide.

The structure-activity relationships for vasoactive intestinal peptide (VIP) receptor binding were studied using N-terminally modified VIP analogs. VIP fragments, and VIP receptor antagonists. Tissue sources included bovine coronary artery, rat mesenteric artery, rat pituitary, rat brain synaptosomes, and rat liver. Experimental conditions for receptor binding were maintained as near to identical as possible. The competitive binding curves for VIP analogs were similar in the bovine and rat vascular preparations. However, appreciable differences were observed between the vascular and other preparations. The vascular receptors discriminated between [D-His1]VIP and [Phe1]VIP, whereas the receptors in other tissues did not. The greatest selectivity was found for [D-Ala4]VIP, which was among the lowest affinity analogs tested on the vasculature but among the highest affinity analogs in the other preparations. The rank orders of analog potencies were comparable for the rat brain and pituitary receptors. The rat liver VIP receptor differed from its counterpart in brain and pituitary predominantly by discriminating between [D-Phe2]VIP and [D-Arg2]VIP. The two VIP receptor antagonists bound weakly and nonselectively to all receptor preparations. Integrity of the full VIP molecule was necessary for full potency of binding to the vascular receptor. We conclude that the vascular VIP receptor possesses recognition properties that are distinct from those for VIP receptors in liver, pituitary, or brain.

Animals↗
PubMed1990

In vitro effects of retinoic acid.

Retinoids, synthetic and natural analogues of vitamin A, play fundamental roles both in directing the spatial organization of cells during the development of vertebrate limbs and in the maintenance of growth and differentiation of many adult tissues. They also block the phenotypic expression of cancer in vitro; inhibit growth and induce differentiation in many animal and human malignant cell types. They have proved beneficial in skin diseases, cancer prevention and in acute promyelocytic leukemia.

Animals↗
PubMed1990

[Effect of wuwei chongji on the mechanical activity of the stomach of SD rats in vivo].

The effect of Wuwei Chongji on the mechanical activity of stomach of SD rats was studied using fixed point method. The experimental results showed that Wuwei Chongji increased the amplitude and frequency of contraction of stomach in rats, and was able to adjust the high and low tension of contraction of stomach. The contraction of the stomach was not influenced by Natrii Chloridi (0.9%) at the same dose.

Animals↗
PubMed1989

Immunodeficiency and clonal growth of target cells induced by helper-free defective retrovirus.

The murine acquired immunodeficiency syndrome is induced by a defective retrovirus. To study the role of virus replication in this disease, helper-free stocks of defective Duplan virus were produced. These stocks were highly pathogenic in absence of detectable replicating murine leukemia viruses (MuLVs) other than xenotropic MuLV. They induced expansion of the infected cell population (over 1000-fold), and this cell expansion was oligoclonal in origin and, most likely, arose through cell division. These results suggest that this defective virus is oncogenic, inducing a primary neoplasia associated with an acquired immunodeficiency syndrome as a paraneoplastic syndrome. These data emphasize the need to determine whether virus replication is necessary for the progression of other immunodeficiency diseases, including acquired immunodeficiency syndrome, and whether these diseases also represent paraneoplastic syndromes.

Animals↗
PubMed1989

Levels of protein kinase C activity in human gastrointestinal cancers.

The protein kinase C (PKC) activities of tumor tissue and adjacent normal mucosa of human cancers of the esophagus (8 cases), stomach (1 case) and colon (3 cases) were measured. Considerable variations were found in the activity of PKC and in its subcellular distribution in these cancers. The PKC activities of the membrane and cytosolic fractions of the eight esophageal cancers were, however, similar to those of the adjacent normal mucosa: the average PKC activities of the tumor tissues and normal mucosa were 7.5 and 8.3 pmol/min/mg protein, respectively, in their membrane fractions and 7.9 and 7.8 pmol/min/mg protein, respectively, in their cytosolic fractions.

Cell Membrane↗
PubMed1989

Epidermal growth factor induces activation of protein kinase FA and ATP.Mg-dependent protein phosphatase in A431 cells.

The cytosolic fractions from epidermal growth factor (EGF)-treated A431 cells exhibit a marked increase in activities of ATP.Mg-dependent protein phosphatase and its activating factor (protein kinase FA) when compared to controls in the absence of EGF. By contrast, the Triton X-100-solubilized membrane fractions from the same EGF-treated cells exhibit a corresponding decrease in protein kinase FA activity. The EGF-dependent activation of protein kinase FA and ATP.Mg-dependent protein phosphatase occurred within physiological concentrations of EGF (ED50 = 5 x 10(-10) M). The changes of kinase and phosphatase activities which were measured concomitantly exhibit very similar characteristics as to EGF sensitivity and time dependence. The EGF-induced kinase and phosphatase activation occurred very rapidly, reaching the maximal activity levels within 3 min. Moreover, the EGF effect is transient; both EGF-stimulated phosphatase and kinase activities returned to control levels within 30 min. Taken together, the results suggest that EGF may induce the activation of kinase FA in the membrane and thereby promotes the activation of ATP.Mg-dependent phosphatase in the cytosol. Exposure of A431 cells to exogenous phospholipase C also resulted in the activation of endogenous kinase FA and ATP.Mg-dependent phosphatase in a similar pattern produced by EGF. This further suggests that phospholipase C can mimic EGF to mediate the activation of kinase FA and ATP.Mg-dependent phosphatase in A431 cells. By its dual role as a multisubstrate protein kinase and as an activating factor of multisubstrate protein phosphatase, protein kinase FA may represent a transmembrane signal of EGF.

Adenosine Triphosphate↗
PubMed1989

The human cytomegalovirus major immediate early promoter can be trans-activated by adenovirus early proteins.

We have examined the effect of adenovirus E1 proteins on expression from the immediate early (IE) region of the human cytomegalovirus (HCMV). The major immediate early promoter, responsive to trans-activation during the HCMV lifecycle, is also responsive to E1 a protein encoded by the 13 S message. E1a proteins inhibit SV40 expression through the mechanism of enhancer repression; however, the presence of E1a proteins did not inhibit expression of the IE region of HCMV. The ability of trans-activate the major IE promoter in the presence of a strong enhancer suggests adenovirus can activate transcription of HCMV upon coinfection. E1b proteins increased levels of steady state mRNA transcribed from the IE region. Increases in expression due to E1a and E1b proteins were additive. These results suggest that adenovirus early expression can activate quiescent HCMV sequences.

Adenovirus Early Proteins↗
PubMed1989

Effects of GTP analogs and dithiothreitol on the binding properties of the vascular vasoactive intestinal peptide receptor.

Previous studies have demonstrated a specific vascular receptor for the neurotransmitter peptide, vasoactive intestinal peptide (VIP), and have suggested that the receptor is positively coupled to vascular adenylate cyclase. The present study addressed the questions whether the vascular VIP receptor is subject to regulation by guanine nucleotides and whether a disulfide reducing agent, dithiothreitol, would perturb the binding function of the vascular VIP receptor. Guanosine triphosphate (GTP) and its non-hydrolyzable analogs, guanylyl imidodiphosphate (Gpp(NH)p) and guanosine-5'-O-(3-thiotriphosphate) (GTP-gamma-S), increased the rate of dissociation of radiolabeled VIP from arterial receptors in a concentration-dependent manner. GTP-gamma-S increased the equilibrium dissociation constant (KD) of the high affinity vascular VIP binding site, a result consistent with decreased high affinity binding of VIP induced by GTP-gamma-S. These results are consistent with a regulatory role for guanine nucleotides in the function of the vascular VIP receptor. The disulfide reducing agent, dithiothreitol, caused a decrease in specific binding of radiolabeled VIP. Upon Scatchard analysis the effect of dithiothreitol was characterized by an increase in the KD and a decrease in the maximum number of binding sites (Bmax) of the high affinity binding site. These results suggest that disulfide bonds are important for ligand binding to vascular VIP receptors. The sulfhydryl alkylating agents, N-ethylmaleimide and iodoacetamide, had minimal effects on radioligand binding.

Animals↗