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Biomedical subjects

H Y Yang

Publications and source records attributed to H Y Yang.

At least 109 records · Page 6Linked to original sources

Antigenic cross-reactions among immunoglobulin of diverse vertebrates (elasmobranchs to man) detected using xenoantisera.

1. Antisera raised in rabbits and goats against intact immunoglobulins or their constituent light and heavy chains from man, mouse and the galapagos shark (Carcharhinus galapagenesis) were tested for their reactivity with immunoglobulins of elasmobranchs, other lower vertebrates and eutherian and prototherian mammals. 2. Xenoantisera directed against human heavy chain isotypes allowed the serological identification of IgM and IgG immunoglobulins in the echidna (Tachyglossus aculeatus), a monotreme which is one of the most primitive species of extant mammals. 3. The antisera to heavy chains reacted to varying degrees with purified immunoglobulins of non-mammalian species, including the chicken, teleost fish and elasmobranchs in a fashion that was specific for immunoglobulins, but was not related to defined human isotypic markers. 4. The reactions of some antisera seem to skip species known to possess homologous immunoglobulins. 5. Antisera directed against isotypic markers of human kappa and lambda light chains reacted with shark light chains in a manner that was specific for light chain determinants but was not isotype-related. 6. Antisera directed against heavy chains of either sharks or mammals reacted with heavy chains, but not with light chains of diverse species. 7. A rabbit antiserum specific for shark light chain reacted with human and murine monoclonal lambda chains and with two synthetic peptides corresponding to human V lambda Fr3 and Fr4 sequences. 8. These results establish that a variety of antigenic markers including conformational and linear determinants can be shared among immunoglobulins of vertebrates species that had an ancestral divergence more than 400 million years ago.

Amino Acid Sequence↗

Adenosine dephosphorylates myosin light chain in primary cultures of vascular smooth muscle cells from normotensive and spontaneously hypertensive rats.

OBJECTIVE: The aim was to determine whether vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR) respond differently to adenosine than those from normotensive Wistar-Kyoto (WKY) rats. DESIGN: Confluent primary cultures of VSMC derived from SHR and WKY aorta and mesenteric arteries and cerebral arteries were used. The effect of adenosine upon cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) formation and the phosphorylation of myosin light chain (MLC) was studied. METHODS: MLC phosphorylation was estimated by subjecting VSMC extracts incubated with 32P to gel electrophoresis, followed by autoradiography and laser densitometry. cAMP and cGMP levels were measured by radioimmunoassay. RESULTS: Baseline MLC phosphorylation levels were not significantly different in SHR and WKY VSMC. Adenosine caused dephosphorylation of MLC in a time- and dose-dependent manner. A maximal response of approximately 40% below control values was observed 5 min after addition of 10(-5) mol/l adenosine in SHR and WKY VSMC with no significant difference between the two strains. The maximally effective concentration of 10(-5) mol/l adenosine evoked increases in both cAMP and cGMP in VSMC from SHR and WKY rats to the same degree. CONCLUSION: We conclude that the overall ability of VSMC to relax, as evidenced by a marked decrease in MLC phosphorylation in response to adenosine, is unaltered in SHR.

Adenosine↗

Distinct roles for the two cGATA-1 finger domains.

We have generated and analyzed by functional assays mutations of the chicken erythroid transcription factor GATA-1. The cGATA-1 protein contains two related finger domains highly conserved across species and characteristic of the family of GATA-binding factors. We find that mutations in the C-terminal finger or adjacent basic region abolish sequence-specific DNA binding, confirming that this region constitutes a novel DNA-binding domain sufficient to recognize the consensus WGATAR motif. At least three separate regions outside of this finger II domain contribute in a cooperative manner to the trans-activation potential of the protein. As expected from previous results analyzing the mouse homolog, we find that the N-terminal finger plays a role in DNA binding by affecting the stability of the DNA-protein complex. In addition, we find mutations of finger I subtly altered in DNA-binding function which greatly diminish trans-activation. Our results support the notion that the GATA-1 protein must be positioned precisely on the GATA cis element to enable the activation of target genes.

Animals↗

Sensorineural deafness inherited as a tissue specific mitochondrial disorder.

We present here a large Israeli-Arab kindred with hereditary deafness. In this family 55 deaf subjects (29M, 26F), who are otherwise healthy, have been identified and traced back five generations to one common female ancestor. The deafness is progressive in nature, usually presenting in infancy and childhood. Audiometry on six deaf and seven unaffected subjects was consistent with severe to profound sensorineural hearing loss. Based on formal family segregation analysis, the inheritance of deafness in this family closely fits the expectation of a two locus model owing to the simultaneous mutation of a mitochondrial gene and an autosomal recessive gene. Thus, this disorder appears to have the unusual features of being an inherited tissue specific mitochondrial disease and apparently requiring the homozygous presence of a nuclear gene for clinical expression. Most importantly, this disorder presents a unique opportunity to investigate the molecular basis of hereditary non-syndromic deafness and normal hearing.

Adult↗

[Study on localization of cystic fluid antigen of Cysticercus cellulosae using McAb-IGSS].

A study was undertaken on the localization of specific combination of McAb against cystic fluid antigen of Cysticercus cellulosae by immunogold silver staining method (IGSS). It was shown that the combination sites of McAb F3 were distinct in the frozen and paraffin sections of Cysticercus cellulosae. The black granules in the bladder walls presented double-layer distribution and those in the scolex folded sites concentrated into pieces. The combination sites of McAb labeled by IGSS against cystic fluid antigen of Cysticercus cellulosae were consistent with the distribution of the cell layers of Cysticercus cellulosae. The sections in the control group had scarce and irregular distribution, which could be differentiated distinctly from those in the experimental group.

Animals↗

IgG from neuropeptide FF antiserum reverses morphine tolerance in the rat.

Previous studies suggest that neuropeptide FF (NPFF) plays a role in opiate dependence and subsequent abstinence syndrome. The present study assessed the role of NPFF in opiate tolerance. Third ventricular injection of IgG from NPFF antiserum restored the analgesic response to i.c.v. morphine in morphine-tolerant rats (radiant heat tail flick test). IgG from control serum failed to produce this effect. In opiate-naive rats, however, the same treatment with IgG from NPFF antiserum did not affect the analgesic response to i.c.v. morphine. Thus, immunoneutralization of NPFF appears to selectively restore morphine sensitivity in opiate-tolerant animals. These results support the hypothesis that endogenous NPFF contributes to opiate tolerance.

Amino Acid Sequence↗

Organogenesis-stage cytochrome P450 isoforms: utilization of PCR for detection of CYP1A1 mRNA in rat conceptal tissues.

Utilizing the reverse transcriptase-linked polymerase chain reaction, we analyzed the capacity of three groups of rat conceptal tissues to express cytochrome P4501A1 (CYP1A1) mRNA during the dysmorphogenesis-sensitive stage of organogenesis. The visceral yolk sac, ectoplacental cone and embryo proper each were investigated on day 12 of gestation with and without prior exposure in utero to 3-methylcholanthrene as inducing agent. With two sets of discriminating oligonucleotide primers, definitive, reproducible signals were detectable only in tissues from 3-methylcholanthrene preexposed conceptuses. Signals of highest intensity were observed with visceral yolk sac tissues and signals of lowest intensity were observed with tissues of the embryo per se. Specificities of the amplified cDNAs were verified using Southern blotting with hybridization to an internal oligonucleotide probe. The results indicate that organogenesis-stage conceptual tissues of the rat will express CYP1A1 mRNA in response to environmental transregulating agents.

Animals↗

Ontogeny of the F8Famide-like (morphine-modulating) peptides in the central nervous system of rats.

FLFQPQRF-NH2 (F8Famide; FMRFamide-like peptide; morphine-modulating peptide) is a peptide isolated from bovine brain. It has some opiate analgesia modulating effects. In an adult rat central nervous system, F8Famide-like immunoreactivity is found in high concentrations in the posterior pituitary, hypothalamus, pons, medulla, and dorsal spinal cord. By using immunohistochemistry and radioimmunoassay, we studied the development of the F8Famide-immunoreactive system in the rat central nervous system during the ontogeny. F8Famide-immoreactive fibers and terminals first appeared in the median eminence on the twentieth embryonal day. Postnatally the F8Famide-immunoreactive system developed rapidly both in the brain and spinal cord, the immunoreactive structures having an adultlike distribution by the age of 1 week. A transient increase of the F8Famide-immunoreactive material was observed during the third and fourth postnatal weeks. By the age of 4 weeks, the F8Famide-containing neuronal system was adultlike both in distribution and quantity. The results suggest that the F8Famide-like peptides may act as neurotransmitters or neuromodulators from the time of their appearance in the brain and spinal cord. The early appearance and the distribution of the F8Famide-like immunoreactivity suggest that these peptides may participate, in addition to nociceptive mechanisms, in the regulation of blood pressure, feeding behaviour, and endocrine functions.

Amino Acid Sequence↗

cAMP-dependent regulation of P450-catalyzed dealkylation in rat conceptal tissues.

Exposures of cultured whole rat conceptuses to varying concentrations of dibutyryl cyclic AMP or isobutylmethylxanthine, alone or in combination, resulted in significant increases in rates of cytochrome P450-dependent depentylation of pentoxyphenoxazone in cell-free preparations. Lesser increases in rates of debenzylation of benzyloxyphenoxazone were also observed. In cultured whole conceptuses, basal depentylase and debenzylase activities in the visceral yolk sac were approximately sixfold higher than in the embryo. The ectoplacental cone and decidual tissues exhibited no detectable depentylase activity. Only the visceral yolk sac exhibited increased depentylase activity in response to dibutyryl cyclic AMP and isobutylmethylxanthine. Inhibitory antibodies raised against adult hepatic P450s IIB1, IIC11, and IA1 failed to significantly inhibit the yolk sac depentylase activities of noncultured conceptuses. The results suggested that the conceptal depentylation reaction may be catalyzed by a unique P450 isoform(s) that is not expressed during adult life.

1-Methyl-3-isobutylxanthine↗

Cytochrome P450-dependent bioactivation of prodysmorphogens in cultured conceptuses.

These investigations were undertaken to determine the extent to which tissues of cultured rat conceptuses contain cytochrome P450 isoforms in sufficient quantities to significantly influence the capacity of certain chemicals to elicit dysmorphogenic effects in vitro. Investigations with highly sensitive probe substrates/inhibitors and with immunologic methods enabled the detection of at least four separate P450 isoforms in tissues of the visceral yolk sac, ectoplacental cone, and embryo proper. One of the isoforms was identified as P450IA1 and was found to be inducible by polycyclic aromatic hydrocarbons in all three tissues. Other isoforms exhibited properties differing from characterized adult rat hepatic isoforms. Each of the isoforms was detectable in conceptuses on gestational days 10, 11, 12, and 14 and was present in the highest concentrations in the visceral yolk sac. Conceptal P450IA1 catalyzed the conversion of dysmorphogenically inactive 2-acetylaminofluorene to 7-hydroxy-2-acetylaminofluorene, a proximate dysmorphogen. Investigations with microinjections suggested that visceral yolk sac hydroxylation was largely responsible for the bioactivation reaction in vitro. The same isoform exhibited no capacity to influence the dysmorphogenic activity of cyclophosphamide. The results demonstrated that tissues of cultured rat conceptuses may contain P450 isoforms in sufficient amounts to markedly influence the dysmorphogenic activity of substrates of the corresponding isoforms.

Animals↗

Angiotensin II induced phosphorylation of myosin light chain in vascular smooth muscle cells from spontaneously hypertensive and normotensive rats.

STUDY OBJECTIVE: The aim was to determine the changes in the phosphorylation of myosin light chain induced by angiotensin II in cultured vascular smooth muscle cells derived from normotensive (WKY) and spontaneously hypertensive rats (SHR). DESIGN: Extracts of vascular smooth muscle cells incubated with [32P]orthophosphoric acid were subjected to 4M urea-SDS electrophoresis, followed by autoradiography and laser densitometry. EXPERIMENTAL MATERIAL: Confluent primary cultures of vascular smooth muscle cells from aorta, superior mesenteric arteries and cerebral arteries were used. MEASUREMENTS AND MAIN RESULTS: The basal myosin light chain phosphorylation of SHR did not differ significantly from that of WKY. Stimulation with 1 nM angiotensin II increased incorporation of 32P into the myosin light chain, which peaked at 4 min and then slowly declined until 15 min. Exposure to angiotensin II (0.001-10 nM) for 4 min evoked a dose dependent increase in the phosphorylation of myosin light chain with a maximal response 40-45% above basal values. No significant differences in the response to angiotensin II were detected between cells derived from the two strains. Saralasin, a specific angiotensin II antagonist, did not affect the basal phosphorylation of myosin light chain but completely abolished the effect of angiotensin II. CONCLUSIONS: Angiotensin II enhances the phosphorylation of the myosin light chain from vascular smooth muscle cells in aorta, mesenteric arteries, and cerebral arteries, but there are no differences in response between SHR and WKY.

Angiotensin II↗

Reconstitution of the filamentous backbone of lens beaded-chain filaments from a purified 49kD polypeptide.

The beaded-chain filaments unique to the fiber cells of the crystalline lens are composed of a linear array of spheroidal particles which appear to be connected by a filamentous backbone. In order to determine the existence of the putative backbone and to characterize its constituents, one of the major proteins associated with beaded-chains in the chicken lens was investigated. 49kD was isolated in an enriched fraction derived from the 8M urea extract of the lens cell water-insoluble residue. The polypeptide (which exists in several charge isoforms, the major at pI 5.2) was purified sequentially by gel filtration on Sephacryl S-200, hydrophobic interaction chromatography on phenyl-Sepharose, and anionic exchange chromatography on Mono Q, all under denaturing conditions. Immunoblot analyses established that 49kD was immunologically distinct from vimentin, actin, and tubulin/MAPs (representing the three classes of cytoplasmic filaments), as well as from the crystallins. Amino acid analyses demonstrated compositional differences for 49kD compared with lens actin and vimentin, and one- and two-dimensional peptide mapping of 49kD and vimentin revealed no homology. Electron microscopy demonstrated that short, contorted filaments were produced upon removal of purified 49kD from urea to low-salt buffers. In the presence of physiological salt concentrations 49kD assembled into extensive 4-6nm diameter, straight filaments similar to the backbone seen in native beaded-chain filaments, but morphologically distinct from the other cytoplasmic filament classes.

Amino Acids↗

Expression and distribution of cytoskeletal IFAP-300kD as an index of lens cell differentiation.

By their implication in the organization of the intermediate filament (IF) cytoskeleton, IF-associated proteins (IFAPs) can delineate subsets of the same IF type within a cell; moreover, they are proving useful as markers of the differentiation states of certain cells. For these reasons the expression of the vimentin-associated IFAP-300kD was investigated in the constantly differentiating cell lineage of the adult bovine lens. Immunofluorescence microscopy and immunoblot analysis were employed using a monoclonal anti-IFAP-300kD and a rabbit anti-lens vimentin. Cultures of adult lens epithelial cells were immunopositive for the IFAP. By double-label studies the IFAP-300kD pattern co-localized with that of the vimentin-type IF; moreover, the IFAP pattern co-distributed with that of both colchicine-sensitive and -insensitive IF systems. IFAP-300kD was also present in a co-distributing pattern with vimentin IF in fresh lens epithelial cells on whole mounts. There was a differential expression of the IFAP in the lens fiber cells in that those of the cortex exhibited the IFAP and vimentin IF, while both proteins were absent from the nuclear fiber cells. Furthermore, there was a differential distribution of the IFAP within the cortical fiber cells in that the IFAP localized only with a paramembranal subset of IF. Immunoblot analysis supported the presence of IFAP-300kD in the lens cytoskeletal fraction. IFAP-300kD thus identified a subset of vimentin IF whose location may have functional significance for the cortical fiber cell. The changes in the IFAP's expression and distribution pattern throughout lens cell differentiation in the adult organ suggest the usefulness of IFAP-300kD as a potential marker in studying lens cell differentiation in vitro.

Animals↗

Histopathological and ultrastructural changes of lichen sclerosus et atrophicus on the vulva.

Studies of 82 skin specimens from 81 cases of lichen sclerosus et atrophicus (LSA) are reported. Typical LSA pathological changes showed atrophy of the epidermis, hydropic degeneration at the basal layer, and homogenization of the collagen fibers in the superficial dermis with inflammatory cell infiltration underneath. Ultrastructural changes showed intercellular spaces enlarged, particularly near basal cells. Some mitochondria were swollen and rough endoplasmic reticula were enlarged. Some basement membranes were thinner, collagen fibers of varying diameters were arranged in irregular shapes and the fine structures had become unclear and had sometimes even disappeared.

Adolescent↗

Further investigation on nuclear transplantation in different orders of teleost: the combination of the nucleus of Tilapia (Oreochromis nilotica) and the cytoplasm of Loach (Paramisgurnus dabryanus).

The nucleus of a blastula cell from Tilapia (Oreochromis nilotica, family Cichlidae, order Perciformes) was transplanted into an enucleated egg of Loach (Paramisgurnus dabryanus, family Cobitidae, order Cypriniformes). From among 3747 nucleo-cytoplasmic hybrid (NCH) eggs two NCH larval fish (0.05%) were obtained; one died on the 6th day and the other died on the 12th day after the operation. Morphological examinations showed that both NCH larval fish had developed normally with an opened mouth except they could not take food after complete utilization of their egg yolk on the 5th day of development. The possible mechanisms for obtaining such inter-order NCH larval fish are discussed. This is the first report indicating that inter-order NCH larval fish can be obtained in spite of their evolutionary divergence.

Animals↗

Prevalence of Toxoplasma gondii infection in man and animals in Guangdong, Peoples Republic of China.

Tissue and serum samples from animals and man in Guangdong Province of the Peoples Republic of China were examined for Toxoplasma gondii infection. Tissues from 519 swine, 576 rodents, 84 people, one cat and two dogs were bioassayed in mice. T. gondii was isolated from 13 pools of swine tissues, but not from any other hosts. Serum samples from animals and man were examined at 1:64 dilution in the indirect hemagglutination test. Antibodies to T. gondii were found in 10.4% of 816 pigs, 0.9% of 955 rodents, 0.7% of 3085 people, 4.4% of 90 cattle, 8.3% of 12 rabbits and 2.1% of 47 cats. None were found in 83 buffaloes.

Animals↗