Search PubMed⌕ Search

Biomedical subjects

H Yuan

Publications and source records attributed to H Yuan.

At least 199 records · Page 11Linked to original sources
PubMed1989

Reaction of rat liver DT-diaphorase (NAD(P)H:quinone acceptor reductase) with 5'-[p-(fluorosulfonyl)benzoyl]-adenosine.

Rat liver DT-diaphorase is inactivated by 5'-[p-(fluorosulfonyl)benzoyl]adenosine (5'-FSBA), following pseudo-first-order kinetics. A double-reciprocal plot of 1/kobs versus 1/[5'-FSBA] yields a straight line with a positive y intercept, indicative of reversible binding of the inhibitor before an irreversible incorporation. The dissociation constant (Kd) for the initial reversible enzyme-inhibitor complex is estimated at 2.86 mM with k2 = 0.22 min-1 (at pH 7.5 and 25 degrees). A stoichiometry of 2 mol of 5'-FSBA bound/mol of enzyme (i.e., 1 mol of the inhibitor bound/mol of subunit), at 100% inactivation, was determined from inactivation kinetics and from incorporation studies using 5'-[p-(fluorosulfonyl)benzoyl]-[14C]-adenosine. The irreversible inactivation as well as the covalent incorporation could be completely prevented by the presence of NAD(P)H during the incubation. These results indicate that 5'-FSBA inactivated DT-diaphorase by occupying its NAD(P)H binding site. Reverse phase high pressure liquid chromatography of tryptic digests of [14C]5'-FSBA-labeled DT-diaphorase revealed one radioactive peak containing two comigrating peptides. They are 146I-T-T-G-G-S-G-S-M-Y155 and 262S-I-P-A-D-N-Q-I-K270. By comparison of these sequences to those of the nucleotide binding sites of several kinases and dehydrogenases, it is suggested that the peptide I-T-T-G-G-S-G-S-M-Y is the one modified by 5'-FSBA and would be predicted to be the region where the pyrophosphate group of NAD(P)H binds.

Adenosine↗
PubMed1988

Structure-function relationship of NAD(P)H:quinone reductase: characterization of NH2-terminal blocking group and essential tyrosine and lysine residues.

The amino terminal blocked peptide of rat liver NAD(P)H:quinone reductase (DT-diaphorase) was determined by amino acid sequence analysis and by mass spectrometry. The mature protein is composed of 273 amino acids and contains an acetylated amino terminus, which was not identified by previous cDNA analysis. The enzyme was inactivated by p-nitrobenzenesulfonyl fluoride (NBSF) or 2,4,6-trinitrobenzenesulfonate (TNBS) with pseudo-first-order kinetics. These studies suggest that essential tyrosine and lysine may be present in the active site of this enzyme. The NBSF inhibition was protected by 1-naphthol and 1-naphthylamine, but not by NAD+. However, TNBS inhibition was not prevented by the naphthalene derivatives or NAD+. Specific peptides labeled with NBSF or TNBS were isolated by high-performance liquid chromatography and were sequenced. These analyses revealed that the NBSF-labeled tyrosine resides in a predominantly hydrophobic region and TNBS-labeled lysine in a predominantly hydrophilic region.

Amino Acid Sequence↗
PubMedDate not supplied

Increased susceptibility of late passage human diploid fibroblasts to oxidative stress.

Three strains of human diploid fibroblasts, TIG-3, TIG-7, and MRC-5, were serially cultivated. The susceptibility of early-passage and late-passage cells at 20-30 and 60-70 population doubling levels, respectively, to hydrogen peroxide, the superoxide radical (exposure to the hypoxanthine-xanthine oxidase system), or linoleic acid hydroperoxide was examined for lactate dehydrogenase release. The susceptibility of late-passage cells to such oxidative stress was considerably enhanced compared with early-passage cells. The concentration of reduced glutathione in late-passage cells was lower by 24-44% on a per-cell-number basis and by 86.0-94.5% on a per-protein-quantity basis than in early-passage cells. In addition, the activity of catalase in late-passage cells was lower by 19-46% compared with early-passage cells. There was, however, no difference between the mRNA levels of catalase in early-passage and late-passage cells. The activities and mRNA levels of copper/zinc superoxide dismutase, manganese superoxide dismutase, and glutathione peroxidase in late-passage cells were all higher than in early-passage cells. These results suggest that late-passage cells are more susceptible to oxidative stress than early-passage cells presumably because of decreases in cellular reduced glutathione concentration and catalase activity, and that their primary defense against oxidative stress is reduced glutathione.

Catalase↗
PubMedDate not supplied

Psychosocial correlates of nicotine dependence among men and women in the U.S. naval services.

This study assessed the prevalence and psychosocial correlates of clinical criteria-based diagnoses of nicotine dependence in men and women in the U.S. naval services. Analyses were based on data from the 1995 Perceptions of Wellness and Readiness Assessment, a population-based self-report survey of 9,856 active-duty Navy and Marine Corps members worldwide, specifically focusing on a subsample of those who completed the telephone version of the Quick Diagnostic Interview Schedule (n = 782). Twenty-five percent of all telephone interview volunteers met diagnostic criteria for lifetime nicotine dependence, and 15% met criteria for nicotine dependence in the last year. There was no difference in nicotine dependence by job classification (traditional vs. nontraditional) among women. Women who were abused prior to entering the military were almost three times as likely to be nicotine dependent as those not abused. Men who had been exposed to combat or violence were twice as likely to be nicotine dependent as those not exposed.

Adaptation, Psychological↗
PubMedDate not supplied

Topological indices based on vertex, distance, and ring: on the boiling points of paraffins and cycloalkanes.

Vertex, distance, and ring (in cyclic compounds) are three essential structure elements of a molecular graph, based on which three new topological indices VDI, OEI, and RDI are proposed. Multiple regression analysis was carried out against the boiling points of 343 hydrocarbons(160 paraffins and 183 cycloalkanes) with VDI, OEI, RDI, and N(2/3)(N is the number of carbon atoms) together, and a good QSPR model was obtained: ln(577 - Bp) = 6.729609 - 0.154107N(2/3) + 2.285632 x 10(-2)VDI - 7.921410 x 10(-3)OEI - 1.821962 x 10(-2)RDI (F = 6455.09, r = 0.9935, rms = 6.44 degrees C, n = 343).

Journal Article↗
PubMedDate not supplied

Measurement of vitellogenin gene expression by RT-PCR as a tool to identify endocrine disruption in Japanese medaka (Oryzias latipes).

In order to monitor vitellogenin gene expression in the Japanese medaka (Oryzias latipes), a reverse transcription-polymerase chain reaction (RT-PCR) system was developed. To date cDNA for medaka vitellogenin has not been published; therefore, initially a sequence fragment had to be obtained and compared with other known vertebrate vitellogenins. For this, a 1.2 kb cDNA of medaka vitellogenin (M-Vg1.2) was amplified by RT-PCR and cloned into a pCRR H-TOPO bacterial vector. On Northern blot analysis, the antisense cRNA of M-Vg1.2 stained a 5.5 kb gene product found exclusively in female fish, but not in males. Additionally, the 5'-end of medaka vitellogenin cDNA was amplified by 5'-RACE-PCR. The analysed nucleotide sequence of 1.6 kb shared significant similarities with vitellogenins known from other fish species: approximately 72% similarity with mummichog (Fundulus heteroclitus) vitellogenin I and approximately 62% with fathead minnow (Pimephales promelas) vitellogenin. To develop a semiquantitative RT-PCR for the measurement of vitellogenin gene expression, primers specific to a 500 bp sequence of the vitellogenin cDNA (M-Vg0.5) were constructed using the gene product of elongation factor 1 alpha as internal standard. Induction of vitellogenin gene expression was measured in male medaka exposed to 0, 2, 20 and 50 micrograms l-1 nonylphenol and 0, 2.5, 25 and 100 ng l-1 17 alpha-ethinyloestradiol for 7 days. The LOECs for vitellogenin induction in male medaka were 20 micrograms l-1 and 25 ng l-1 for nonylphenol and 17 alpha-ethinyloestradiol, respectively.

Animals↗
PubMedDate not supplied

Automated in-tube solid-phase microextraction coupled with liquid chromatography-electrospray ionization mass spectrometry for the determination of selected benzodiazepines.

A simple, rapid, and sensitive method, which allowed us to simultaneously determine seven benzodiazepines (diazepam, nordiazepam, temazepam, oxazepam, 7-aminoflunitrazepam, N-desmethylflunitrazepam, and clonazepam) in buffer solution and in urine and serum samples, was investigated by automated in-tube solid-phase microextraction (SPME) coupled with liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS). In-tube SPME, in which the analytes were extracted from the sample directly into an open tubular capillary column by repeated draw/eject cycles of sample solution, is an extraction technique for organic compounds in aqueous samples. The separation of benzodiazepines was carried out under ion-suppressed reversed-phase conditions by using methanol/50mM ammonium acetate in water (60:40) as a mobile phase with a Supelco LC-18 column. The optimal extraction condition was 10 draw/eject cycles of 30 mL of sample in 100mM Tris-HCl (pH 8.5) at a flow rate of 0.3 mL/min using a piece of 60-cm length Supelco-Q plot capillary column as the extraction capillary. The quantitative study was explored by operating in selected-ion monitoring (SIM) mode. The calibration curves were linear in the range from 0.5 ng/mL or 2 ng/mL to 500 ng/mL. The detection limits were from 0.02 ng/mL to 2 ng/mL. At the optimized capillary and fragmentor voltages, the characteristic ions for each compound clearly showed up in the spectra and it is possible to use the LC-MS to identify these compounds. The method was applied to the analysis of biological samples without interfering peaks. However, the recoveries for some of the compounds in serum samples need to be further improved.

Benzodiazepines↗
PubMedDate not supplied

Lactate elicits vascular endothelial growth factor from macrophages: a possible alternative to hypoxia.

Macrophages respond to various stimuli to produce angiogenic factors but few mechanistic details are known. We examined the effects of hypoxia, lactate and nicotinamide on the expression of vascular endothelial growth factor by cultured macrophages. These agents were chosen because they down-regulate polyadenosine diphosphoribose levels. Following exposure, conditioned media were analyzed for vascular endothelial growth factor protein. Nicotinamide adenine dinucleotide, polyadenosine diphosphoribose, and vascular endothelial growth factor mRNA were measured in the cellular fraction. Angiogenic capacity of the conditioned media was tested in rabbit corneas and Matrigel implants. All three agents, hypoxia, lactate and nicotinamide, elicited significantly increased levels of vascular endothelial growth factor mRNA and vascular endothelial growth factor in the conditioned media, and these levels were paralleled by their angiogenic activity. Polyadenosine diphosphoribose in the cellular fraction was correspondingly depressed. Anti-vascular endothelial growth factor antibody inhibited most of the angiogenic response whereas anti-basic fibroblast growth factor antibody had little effect. We propose that redox changes associated with the alteration of cellular nicotinamide adenine dinucleotide and polyadenosine diphosphoribose are involved in lactate-mediated VEGF expression.

Animals↗
PubMedDate not supplied

Sensitivity analysis for evaluating nonlinear models of lung mechanics.

We present a combined theoretical and numerical procedure for sensitivity analyses of lung mechanics models that are nonlinear in both state variables and parameters. We apply the analyses to a recently proposed nonlinear lung model which incorporates a wide range of potential nonlinear identification conditions including nonlinear viscoelastic tissues, airway inhomogeneities via a parallel airway resistance distribution function, and a nonlinear block-structure paradigm. Additionally, we examine a system identification procedure which fits time- and frequency-domain data simultaneously. Model nonlinearities motivate sensitivity analyses involving numerical approximation of sensitivity coefficients. Examination of the normalized sensitivity coefficients provides direct insight on the relative importance of each model parameter, and hence the respective mechanism. More formal quantification of parameter uniqueness requires approximation of the paired and multidimensional parameter confidence regions. Combined with parameter estimation, we use the sensitivity analyses to justify tissue nonlinearities in modeling of lung mechanics for healthy and airway constricted conditions, and to justify both airway inhomogeneities and tissue nonlinearities during bronchoconstriction. The tools in this paper are general and can be applied to a wide class of nonlinear models.

Airway Resistance↗
PubMedDate not supplied

Parametric and nonparametric nonlinear system identification of lung tissue strip mechanics.

Lung parenchyma is a soft biological material composed of many interacting elements such as the interstitial cells, extracellular collagen-elastin fiber network, and proteoglycan ground substance. The mechanical behavior of this delicate structure is complex showing several mild but distinct types of nonlinearities and a fractal-like long memory stress relaxation characterized by a power-law function. To characterize tissue nonlinearity in the presence of such long memory, we investigated the robustness and predictive ability of several nonlinear system identification techniques on stress-strain data obtained from lung tissue strips with various input wave forms. We found that in general, for a mildly nonlinear system with long memory, a nonparametric nonlinear system identification in the frequency domain is preferred over time-domain techniques. More importantly, if a suitable parametric nonlinear model is available that captures the long memory of the system with only a few parameters, high predictive ability with substantially increased robustness can be achieved. The results provide evidence that the first-order kernel of the stress-strain relationship is consistent with a fractal-type long memory stress relaxation and the nonlinearity can be described as a Wiener-type nonlinear structure for displacements mimicking tidal breathing.

Algorithms↗
PubMedDate not supplied

[125I]iodomelatonin binding sites in the chicken brain: diurnal variation and effect of melatonin injection or pinealectomy.

The diurnal variation of [125I]iodomelatonin binding sites in the brain and serum melatonin levels were studied at 4-hour intervals under a 12 h:12 h light:dark cycle in 5-week-old chicks. There was a diurnal rhythm of [125I]iodomelatonin binding capacities in brain membrane preparations with the peak in the light period and the trough in the dark period. However, a diurnal variation with high levels in the dark and low levels in the light was observed in serum melatonin concentrations. The binding capacity of [125I]iodomelatonin was inversely related to the serum melatonin level. Four-week-old chicks were injected with melatonin (1 mg melatonin/kg; i.p.) twice a day (shortly after the light onset and shortly before light offset) for 3 weeks. When chickens were killed at 7 weeks, there was no diurnal rhythm of melatonin in the circulation. In addition, melatonin injection also abolished the diurnal variation of [125I]iodomelatonin binding sites in brain membrane preparations in these animals. This was achieved by decreasing the binding capacity in the light period. Pinealectomy in 2-day-old chickens increased the density of [125I]iodomelatonin binding sites in the brain membrane preparations both at midlight and middark when the chicks were later sacrificed at 6 weeks of age, although the rhythm of binding sites persisted. It is suggested that the [125I]iodomelatonin binding capacity in the chicken brain is regulated by endogenous melatonin and influenced by exogenous melatonin. In addition, the rhythmicity of [125I]iodomelatonin binding sites in the chicken brain is affected by, but not dependent on, pineal melatonin.

Animals↗
PubMedDate not supplied

Effects of guanosine 5'-O-(3-thiotriphosphate) on 2-[125I]iodomelatonin binding in the chicken lung, brain and kidney: hypothesis of different subtypes of high affinity melatonin receptors.

Effects of 10 mumol/l guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S), a non-hydrolyzable analog of guanosine 5'-triphosphate, on 2-[125I]iodomelatonin binding were investigated. In the chicken lung, 10 mumol/l GTP gamma S significantly increased (p < 0.05) the equilibrium dissociation constant (Kd) values, but did not affect the maximum number of binding sites (Bmax). Conversely, in the chicken brain, GTP gamma S significantly depressed (p < 0.05) the Bmax, but did not change the Kd of the 2-[125I]iodomelatonin binding sites in the brain tissue. A third variation was observed in the chicken kidney with GTP gamma S altering (p < 0.05) both the Kd and the Bmax of 2-[125I]iodomelatonin binding sites. The reason underlying the different effects of GTP gamma S on 2-[125I]iodomelatonin binding in the tissue preparations is not clear. However, we would like to hypothesize that they may represent distinct subtypes of the ML-1-type melatonin receptor with different receptor-G-proteins-effector complex. The group represented by the 2-[125I]iodomelatonin binding sites found in the chicken lung, which is downregulated by GTP gamma S with a consequent increase in the Kd value, has been designated ML-1 alpha. The second group, exemplified by the brain 2-[125I]iodomelatonin binding sites, which respond to GTP gamma S with a change in Bmax, has been labelled ML-1 beta. The third group, characterized by GTP gamma S-mediated alterations in both Bmax and Kd and found in the chicken kidney, has been called ML-1 gamma. Different subtypes of melatonin receptors may address the issue of the different physiological actions of melatonin reported in individual tissues within the same species or similar tissues but different species. Specialized responses could be generated depending on the predominant subtype of ML-1 receptors associated with the target tissue.

Animals↗