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Biomedical subjects

Y Hu

Publications and source records attributed to Y Hu.

At least 577 records · Page 32Linked to original sources
PubMed1994

Role of guanylyl cyclase and cGMP-dependent protein kinase in long-term potentiation.

Several lines of evidence suggest that cyclic GMP might be involved in long-term potentiation (LTP) in the hippocampus. Arachidonic acid, nitric oxide and carbon monoxide, three molecules that have been proposed to act as retrograde messengers in LTP, all activate soluble guanylyl cyclase. We report here that an inhibitor of guanylyl cyclase blocks the induction of LTP in the CA1 region of hippocampal slices. Conversely, cGMP analogues produce long-lasting enhancement of the excitatory postsynaptic potential if they are applied at the same time as weak tetanic stimulation of the presynaptic fibres. The enhancement is spatially restricted, is not blocked by valeric acid (APV), nifedipine, or picrotoxin, and partially occludes LTP. This synaptic enhancement may be mediated by the cGMP-dependent protein kinase (PKG). Inhibitors of PKG block the induction of LTP, and activators of PKG produce activity-dependent long-lasting enhancement. These results suggest that guanylyl cyclase and PKG contribute to LTP, possibly as activity-dependent presynaptic effectors of retrograde messengers.

Action Potentials↗
PubMed1994

Elimination of the acetaminophen interference in an implantable glucose sensor.

Acetaminophen has been one of the most serious electrochemical interferences to oxidase-based amperometric biosensors that measure H2O2. A study was carried out to investigate various polymer materials for their selectivity as the sensor inner membrane. A composite membrane of cellulose acetate and Nafion was found to eliminate acetaminophen and other electrochemical interferences effectively while at the same time maintaining reasonable diffusivity for hydrogen peroxide. The excellent in vivo performance of the sensor was attributed not only to significantly reduced steady-state sensitivity to acetaminophen but also to very slow acetaminophen response. These features, combined with rapid acetaminophen clearance pharmacokinetics, led to the decreased response as demonstrated in the rat.

Acetaminophen↗
PubMed1994

Genetic polymorphism of cytochrome P450. Functional consequences and possible relationship to disease and alcohol toxicity.

The hepatic cytochrome P450 system participates in the oxidative metabolism of numerous endogenous and exogenous compounds. In total several hundred different P450s have been cloned, but it appears that in humans only about 5-10 isoforms account for the major part of drug metabolism. Some of these are polymorphically distributed in the population. Cytochrome P450 2D6 catalyzes the oxidation of over 25 clinically important drugs, eg neuroleptics, antidepressants and lipophilic beta-blockers. Seven % of Caucasians and 1% of Orientals are defective in this enzyme and clearance of drugs metabolized by the enzyme may be substantially decreased in these individuals, with potentially increased risks for side effects caused by the drug treatment. Some individuals are ultrarapid metabolizers and do not achieve therapeutic drug levels at ordinary doses. The molecular genetic basis of these polymorphisms are presented. Methods for genotyping, which can be of predictive value for a more efficient drug therapy, are discussed. Ethanol-inducible cytochrome P450 2E1 (CYP2E1) oxidizes ethanol and acetaldehyde, in addition to over 80 toxicologically important xenobiotics. Furthermore, this isozyme produces reactive oxy radicals which are implicated in the aetiology of alcoholic liver disease. The gene is polymorphic and a mutation in a putative binding site for HNF1, described to affect gene expression, is more rare among subjects with lung cancer as compared to healthy controls. Further studies might give an answer as to whether any of the polymorphic CYP2E1 alleles is associated with the sensitivity to obtain alcoholic liver disease.

Animals↗
PubMed1994

Reduction of acetaminophen interference in glucose sensors by a composite Nafion membrane: demonstration in rats and man.

Amperometric glucose sensors typically monitor the production of hydrogen peroxide generated in the course of the enzymatic oxidation of glucose. At the applied potential necessary to oxidize the peroxide produced, other species are also electroactive and contribute to the signal. Interference of ascorbate or urate has been effectively eliminated, but that resulting from the widely used analgesic acetaminophen is not. The aim of this work was to reduce this interference, which was found to be possible by introducing a membrane constructed of Nafion. We compared the in vitro sensitivity to acetaminophen of five Nafion sensors with that of five non-Nafion sensors with identical glucose sensitivity (2.0 +/- 0.4 vs 1.9 +/- 0.1 nA.mmol-1.l-1, NS): sensitivity to acetaminophen was 12.2 +/- 2.7 vs 30.8 +/- 6.3 nA.mmol-1.l-1, respectively (p < 0.05). These sensors were tested in rats by implanting in each animal one Nafion and one non-Nafion sensors. The in vivo sensitivity to glucose was similar (0.33 +/- 0.09 vs 0.30 +/- 0.05 nA.mmol-1.l-1, NS). The current generated by an acetaminophen infusion (plasma acetaminophen plateau = 140 +/- 10 mumol/l) was much decreased in the case of the Nafion sensor: 0.5 +/- 0.3 vs 2.0 +/- 0.7 nA, p < 0.05). Five Nafion sensors were implanted in the subcutaneous tissue of normal human volunteers who were given on oral dose of 500 mg acetaminophen.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetaminophen↗
PubMed1994

A densovirus newly isolated from the smoky-brown cockroach Periplaneta fuliginosa.

We purified a causing agent of fetal disease for smoky-brown cockroach Periplaneta fuliginosa, which was designated as "cockroach small spherical virus (CSSV)". Purified virus particles had a diameter of 22 +/- 0.6 nm and contained DNA as a single-stranded form. However, the extraction of DNA under condition of appropriate high salt and elevated temperature yielded a double-stranded DNA with a size of 5,500 nucleotides. These results were quite similar to those of other densoviruses (DNVs). The CSSV had five structural proteins (VP1: 52 KDa, VP2: 56 KDa, VP3: 79 KDa, VP4: 82 KDa, and VP5: 105 KDa). The SDS-PAGE profile of these proteins was quite different from that of the cockroach DNV previously reported and was rather similar to that of Bombyx mori (Bm) DNV-1. An immunochemical study, however, demonstrated that there was no immunological relationship between the CSSV and the Bm DNV-1. These data suggest that the CSSV is a new member of DNV.

Animals↗
PubMed1994

Interleukin-1 receptor deficiency in brains from NZB and (NZB/NZW)F1 autoimmune mice.

Interleukin-1 receptors (IL-1R) are expressed in the brain and the anterior pituitary of normal mice (C3H/He, Swiss), and appear to be involved in the neuroendocrine control of the immune response. Here we have studied the IL-1R density in the brain and the pituitary from several strains of autoimmune mice (NZB, (NZB/NZW)F1, MRL/MP-lpr), using quantitative autoradiography with recombinant human [125I]IL-1 alpha as a ligand. IL-1R was similar in the brain of C3H/He, Swiss and NZW (controls) and MRL/MP-lpr mice. In NZB mice a profound deficit (10% of control mice) in IL-1R was observed exclusively in the dentate gyrus. In (NZB/NZW)F1 the deficit was about 50%. These observations were independent of sex and age. Pituitary receptors were not affected in all the strains except NZW (30% increase). Competition experiments demonstrated that the affinity of IL-1R was not modified in dentate gyrus of (NZB/NZW)F1 and NZW mice. Thus, the number of IL-1R was the only parameter affected. This deficit was not reversed by corticosterone treatment (0.2 mg/20 g body weight, i.p.) and was poorly modified by lipopolysaccharide treatment (0.1 mg/20 g body weight, i.p.) compared to C3H/He mice. In conclusion, this central IL-1R deficit is unlikely to be the consequence of occupancy by abnormal synthesis of brain IL-1. This abnormality is tissue-specific with hereditary autosomal transmission. The role of central IL-1R in neuroimmunoendocrine interactions and in autoimmunity remains to be clarified.

Age Factors↗
PubMed1994

Expression of Thy-1/lacZ fusion genes in the CNS of transgenic mice.

Thy-1 is a cell surface glycoprotein of unknown function that is found on nerve cells and mature T-lymphocytes. To study the regulation of Thy-1 gene expression, mouse Thy-1.2 genomic sequences were joined to various marker sequences and the resulting chimeric constructs were used to produce nearly three dozen independent lines of transgenic mice. The starting point for our studies was an 8.2 kb EcoRI fragment that begins 1.7 kb 5' to the transcription start site and ends with 1.3 kb of 3' flanking sequences. Addition of a small marker oligonucleotide to the 3' untranslated region of this fragment had little or no effect on gene regulation. All of the lines derived from injection of this construct expressed the transgene in the appropriate tissues. Thus, as expected, the Thy-1.2 genomic fragment contains all of the information necessary for tissue-specific, position-independent expression of the modified transgene. Unexpectedly, Thy-1/lacZ hybrid genes did not mimic this behavior. Using either mRNA or histochemical detection of lacZ protein, these constructs were expressed in patterns that varied dramatically from line to line. This behavior suggests that integration site-specific effects dominate the cis-active Thy-1 regulatory elements leading to wide variability of expression. This is further emphasized by the observation that the bacterial reporter protein was found in a few non-neuronal cell-types, in contrast to the known pattern of native Thy-1 expression. These results suggest that either the Thy-1.2 sequences which are necessary for appropriate brain-specific expression are not contained solely within the proposed CNS enhancer in the first intron, or that fusion of the Thy-1.2 sequences with the lacZ coding region may disrupt normal Thy-1 regulatory signals (or result in the creation of new regulatory elements).

Aging↗
PubMed1994

The combination of coculture and selective assisted hatching: results from their clinical application.

OBJECTIVE: To establish pregnancies using a combination of coculture and selective assisted hatching. DESIGN: Clinical application for a selected group of patients. Not a controlled study. SETTING: Private infertility practice. PATIENTS: Women with high basal FSH levels, ovulatory disorders, and multiple failed IVF attempts. MAIN OUTCOME MEASURES: Pregnancy and implantation rates. RESULTS: Of the 95 patients who had coculture and selective assisted hatching, 45 (47.0%) have an ongoing pregnancy with a 23.0% implantation rate. CONCLUSION: The combination of coculture and assisted hatching produced acceptable pregnancy and implantation rates within the selected patient population.

Adult↗
PubMed1994

Blockade of ornithine decarboxylase enzyme protects against ischemic brain damage.

Polyamines are derived from ornithine by the actions of ornithine decarboxylase (ODC), which is the rate-limiting step in this pathway. Polyamines play a role in cell growth, neoplasia, differentiation, and response to injury. We have shown that transient cerebral ischemia gives rise to increased ODC mRNA and enzyme activity in the gerbil brain. ODC and polyamines are thought to be important in the generation of edema and the neuronal cell loss associated with cerebral ischemia. To test this theory, we examined the ODC activity, putrescine levels, and neuronal density in the CA1 region of the hippocampus following ischemia and reperfusion injury in the absence and presence of an inhibitor of ODC activity, alpha-difluoromethylornithine (DFMO). Pretreatment of animals with DFMO resulted in attenuation of the ODC activity following 5 min of ischemia and 4 h of reperfusion. In addition, DFMO prevented the increase in polyamine levels, as determined by measurement of putrescine in the ischemic brain. These alterations were not due to changes in ODC mRNA level. Further analysis revealed that DFMO treatment blocked the delayed neuronal cell death in the CA1 region of the hippocampus that accompanies ischemia and reperfusion injury. Administration of DFMO resulted in a dose-dependent beneficial effect upon neuronal cell survival. These results suggest that ODC enzyme activity and the production of polyamines play a significant role in the response of the brain to ischemic injury.

Animals↗
PubMed1994

The nonstructural glycoprotein of rotavirus affects intracellular calcium levels.

Rotavirus infection of monkey kidney cells has been reported to result in a significant increase in the concentration of intracellular calcium. This increase in intracellular calcium was associated with viral protein synthesis and cytopathic effects in infected cells. We tested the effect of individual rotavirus proteins on intracellular calcium concentrations in insect Spodoptera frugiperda (Sf9) cells. Insect cells were infected with wild-type baculovirus or baculovirus recombinants that contained an individual rotavirus gene. The cells were harvested at different times postinfection, and the intracellular calcium concentration was measured by using fura-2 as a fluorescent calcium indicator. We found that the concentration of intracellular calcium was increased nearly fivefold in infected Sf9 cells that expressed the nonstructural glycoprotein (NSP4) of group A rotavirus, and this increase in intracellular calcium concentration coincided with NSP4 expression. A similar result was observed in insect cells expressing NSP4 from a group B rotavirus, suggesting the conservation of this function among rotavirus groups. Expression of the other 10 rotavirus proteins or of wild-type baculovirus proteins in Sf9 cells did not significantly increase intracellular calcium levels. These results suggest that the nonstructural glycoprotein NSP4 is responsible for the increase in cytosolic calcium observed in rotavirus-infected cells.

Animals↗
PubMed1994

Ca2+ signaling in Sf9 insect cells and the functional expression of a rat brain M5 muscarinic receptor.

The purpose of the present study was to examine Ca2+ signaling mechanisms in Sf9 cells and to demonstrate expression and functional linkage of a mammalian receptor to changes in cytosolic free Ca2+ concentration ([Ca2+]i). Addition of p-octopamine (50 microM to fura 2-loaded Sf9 cells produced a small transient increase in [Ca2+]i from a basal level of 58 +/- 10 to 194 +/- 7.6 (SD) nM. The response to octopamine was inhibited by both cyproheptadine and chlorpromazine and was mimicked by clonidine. In contrast, [Ca2+]i did not change in response to dopamine (50 microM), substance P (50 nM), histamine (50 microM), ATP (50 microM), acetylcholine (10 or 100 microM), carbachol (10 or 100 microM), serotonin (50 microM), epinephrine (10 microM), or bradykinin (50 nM). The Ca(2+)-adenosinetriphosphatase inhibitors thapsigargin (200 nM) and 2,5-di-tert-butylhydroquinone (BHQ; 10 microM) increased [Ca2+]i to 307 +/- 13 and 137 +/- 20 nM, respectively. In contrast to BHQ, the response to thapsigargin was attenuated by La3+ or removal of extracellular Ca2+ and increased by elevation of extracellular Ca2+. These results suggest that thapsigargin but not BHQ stimulates Ca2+ influx. The rat brain muscarinic receptor (subtype M5) was incorporated into the baculovirus by homologous recombination. Addition of carbachol (100 microM) increased [Ca2+]i from 92.7 +/- 6.4 to 480 +/- 26 nM in Sf9 cells infected with recombinant virus containing the M5 receptor cDNA. The effect of carbachol on [Ca2+]i was concentration dependent with a 50% effective concentration of approximately 30 microM and was blocked by atropine (10 microM).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Activation of recombinant trp by thapsigargin in Sf9 insect cells.

The mammalian protein responsible for Ca2+ release-activated current (Icrac) may be homologous to the Drosophila protein designated trp. Thus the activity of trp, and another Drosophila protein designated trp-like or trpl, may be linked to depletion of the internal Ca2+ store via the so-called capacitative Ca2+ entry mechanism. To test this hypothesis, the effect of thapsigargin, a selective inhibitor of the endoplasmic reticulum Ca2+ pump, on trp- and trpl-induced whole cell membrane current was determined using the baculovirus Sf9 insect cell expression system. The results demonstrate that trp and trpl form Ca(2+)-permeable cation channels. The trpl encodes a nonselective cation channel that is constitutively active under basal nonstimulated conditions and is unaffected by thapsigargin, whereas trp is more selective for Ca2+ than Na+ and is activated by depletion of the internal Ca2+ store. Although evaluation of cation selectivity suggests that trp is not identical to the channel responsible for Icrac, these channels must share some structural feature(s) since both are activated by thapsigargin. A unique proline-rich region in the COOH-terminal tail of trp, which is absent in trpl, may be necessary for capacitative Ca2+ entry.

Animals↗
PubMed1994

Downregulation of beta-adrenergic receptors and signal transduction response in salivary glands of NOD mice.

The nonobese diabetic (NOD) mouse is subject to autoimmune disease-associated lymphocytic attack on the salivary glands with a corresponding loss of exocrine function. Downregulation of stimulus response to the beta-adrenoceptor agonist, isoproterenol, appears to be related to a decline in beta-adrenergic receptor density, changes in the level of intracellular second messenger signaling component adenosine 3',5'-cyclic monophosphate, and protein kinase A activity. An autoantibody to the beta 1-adrenergic receptor present in the sera of diabetic NOD mice may be involved in the reduced agonist response by virtue of its ability to retard dihydroalprenolol radioligand binding to receptors in the membranes of salivary glands from control mice and recognition of purified beta 1-adrenergic receptor by immunoblotting techniques.

Animals↗
PubMed1994

Surface staining and cytotoxic activity of heat-shock protein 60 antibody in stressed aortic endothelial cells.

Heat-shock protein (hsp) expression can be induced by high temperature, exposure to cytokines or oxygen radicals, ischemia, hemodynamic overload, or viral infections. To determine whether surface expression of hsp60 occurs in aortic endothelial cells stressed by high temperature or cytokines, cells from rat aortas were cultivated and stained with several types of monoclonal antibodies against hsp60. Other antibodies, eg, those against intercellular adhesion molecule-1 (ICAM-1), or immune response-associated antigens were also used as controls. Positive staining of endothelial cells on the surface and in the cytoplasm was observed after pretreatment of the cells with cytokine-containing medium, tumor necrosis factor-alpha (TNF-alpha), or interleukin-1 alpha and labeling with a specific monoclonal antibody against hsp60 (II-13). Fluorescence-activated cell sorter analyses showed that over 80% of living endothelial cells stressed by cytokine-containing medium, by TNF-alpha, or at 42 degrees C, but not by interleukin-1 alpha, were positively surface stained with this antibody. Increased intensity of immunostaining with antibodies to ICAM-1 and immune response-associated antigen was also seen on the cytokine-stressed endothelial cells. Furthermore, when TNF-alpha stimulated endothelial cells labeled with 51Cr were incubated with antibody II-13 in the presence of complement, significant lysis occurred. In summary, endothelial cells stressed by high temperature or certain cytokines, eg, TNF-alpha, express hsp60 in the cytoplasm and on their surfaces, and these cells were susceptible to complement-dependent lysis by hsp60-specific antibody. These observations may be significant for elucidating the mechanisms of the involvement of immune reactions to hsp65/60 in initiating atherosclerosis.

Animals↗
PubMed1994

[Investigation of skull base in nasopharyngeal carcinoma].

100 patients with destruction of skull base in nasopharyngeal carcinoma, confirmed by pathology and CT scanning before radiotherapy, were analysed. It showed that the primary characteristics of skull base destruction are headache and cranial nerve involvement. Although the destruction of skull base showed evident manifestation in TNM classification, it can be inferred that the erosion development probably took place within 1-6 months. Comparing the CT scanning with X-ray submento-vertical view, the result indicated that the erosions of the various positions of the skull base and the invasion of paranasal sinus, cavernous sinus and orbit were clearly displayed by CT scanning but only 23% was discovered by X-ray. There was significant difference between CT scanning and X-ray, and CT scanning had more advantages. The relation between the erosion of the skull base and the invasion of parapharynx space and the metastatics of the cervical nodes were also discussed.

Adolescent↗