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D Han

Publications and source records attributed to D Han.

At least 181 records · Page 10Linked to original sources
PubMed1993

Comparative studies on binding of 3 different ligands to the N-methyl-D-aspartate recognition domain in brain synaptic membranes treated with Triton X-100.

Treatment with a low concentration of Triton X-100 almost tripled the binding of [3H]D,L-(E)-2-amino-4-propyl-5-phosphono-3-pentenoic acid (CGP 39653), a novel competitive antagonist at an N-methyl-D-aspartate (NMDA)-sensitive subclass of brain excitatory amino acid receptors, in synaptic membranes of the rat brain. The binding linearly increased with increasing protein concentrations of up to 0.4 mg/ml and also increased in proportion to incubation time with a plateau within 60 min after the initiation of incubation at 2 degrees C in Triton-treated membranes. Elevation of incubation temperature from 2 degrees C to 30 degrees C resulted in a marked decrease in the binding at equilibrium by 80%, and a maximal level was obtained within 1 min after the initiation of incubation at 30 degrees C with a gradual decline of up to 10 min. Bound [3H]CGP 39653 was rapidly dissociated by the addition of excess unlabeled L-glutamic acid (Glu), and the time required to attain complete dissociation was 60 min at 2 degrees C and 1 min at 30 degrees C, respectively. Among several agonists and antagonists tested, Glu was the most potent displacer of [3H]CGP 39653 binding with progressively less potent displacement by D-2-amino-5-phosphonovaleric, (+-)-3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic (CPP), D-2-amino-7-phosphonoheptanoic, N-methyl-D-aspartic and N-methyl-L-aspartic acids.(ABSTRACT TRUNCATED AT 250 WORDS)

2-Amino-5-phosphonovalerate↗
PubMed1993

Spectroscopic and kinetic studies of lipases solubilized in reverse micelles.

The conformation and activity of three different lipases have been studied in reverse micelles formed by sodium bis(2-ethylhexyl) sulfosuccinate (AOT) in isooctane. In the case of human pancreatic lipase, the conformation of the polypeptide chain--as judged from far-UV circular dichroism measurements--is only slightly altered after the enzyme is transferred from a bulk aqueous solution into the microenvironment of reverse micelles. Significant spectral changes in the near-UV circular dichroism and fluorescence spectrum indicate, however, that the solvation of aromatic amino acid side chains is considerably different in reverse micelles. Conversely, the circular dichroism spectra of the lipases from Candida rugosa and Pseudomonas sp. are considerably different in reverse micelles, compared with the spectra in aqueous solution, indicating that both enzymes loose the native structure at the water/AOT/oil interface. Bound substrate and/or product can prevent this denaturation. While Pseudomonas sp. and human pancreatic lipase are inhibited by tetrahydrolipstatin (THL), the lipase from Candida rugosa is not. These data, together with additional activity and inhibition data, indicate that the micellar microenvironment accentuates the difference between the different enzymes in terms of the relation structure/activity.

Animals↗
PubMed1993

Differential potentiation by spermidine of abilities of a variety of displacers for [3H]MK-801 binding in hippocampal synaptic membranes.

Binding of [3H](+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imi ne (MK-801) to an ion channel associated with the N-methyl-D-aspartate (NMDA)-sensitive subtype of brain excitatory amino acid receptors was studied in Triton-treated preparations of synaptic membranes of rat brain. The initial association rate of the binding measured at 30 min after onset of incubation was markedly potentiated by the addition of either L-glutamic acid (Glu) alone or both Glu and glycine (Gly) in a concentration-dependent manner at 10 nM to 0.1 mM. Potentiation occurred to a significantly greater extent in the hippocampus and cerebral cortex than in the cerebellum. In the presence of both Glu and Gly, the endogenous polyamine spermidine (SPD) further potentiated binding in hippocampal and cortical membranes at concentrations above 10 microM without significantly affecting that in cerebellar membranes. The binding of [3H]MK-801 was slowly equilibrated in 16 h. When examined in hippocampal synaptic membranes, the binding at equilibrium was markedly displaced by numerous noncompetitive antagonists for the NMDA receptor. The addition of SPD markedly enhanced potencies of those displacers having a high affinity to [3H]MK-801 binding sites, without affecting other displacers having a low affinity. These results suggest that SPD promotes transition of sites responsible for mediating NMDA responses within the channel to a state with higher affinity for noncompetitive blockers.

Animals↗
PubMed1993

Support for radiolabeling of a glycine recognition domain on the N-methyl-D-aspartate receptor ionophore complex by 5,7-[3H]dichlorokynurenate in rat brain.

Pretreatment with Triton X-100 more than doubled the binding of radiolabeled 5,7-dichlorokynurenic acid (DCKA), a proposed antagonist at a glycine (Gly) recognition domain on the N-methyl-D-aspartate (NMDA) receptor ionophore complex, in rat brain synaptic membranes. The binding exhibited an inverse temperature dependency, reversibility, and saturability, the binding sites consisting of a single component with a high affinity (27.5 nM) and a relatively low density (2.87 pmol/mg of protein). The binding of both [3H]DCKA and [3H]Gly was similarly displaced by numerous putative agonists and antagonists at the Gly domain in a concentration-dependent manner at a concentration range of 100 nM to 0.1 mM. Among the 24 putative ligands tested, DCKA was the second most potent displacer of the binding of both radioligands with no intrinsic affinity for the binding of [3H]kainic acid and alpha-amino-3-hydroxy-5- [3H]methylisoxazole-4-propionic acid (AMPA) to the non-NMDA receptors. In contrast, the other proposed potent Gly antagonist, 5,7-dinitroquinoxaline-2,3-dione, was active in displacing the binding of [3H]glutamic ([3H]Glu) and D,L-(E)-2-amino-4-[3H]propyl-5-phosphono-3-pentenoic acids to the NMDA recognition domain with a relatively high affinity for the non-NMDA receptors. In addition, the proposed antagonist at the AMPA-sensitive receptor, 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline, not only displaced weakly the binding of both [3H]-Gly and [3H]DCKA, but also inhibited the binding of (+)-5-[3H]methyl-10,11- dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine ([3H]MK-801) to an ion channel associated with the NMDA-sensitive receptor in the presence of added Glu alone in a manner sensitive to antagonism by further added Gly. Clear correlations were seen between potencies of the displacers to displace [3H]DCKA binding and [3H]Gly binding, in addition to between the potencies to displace [3H]-DCKA or [3H]Gly binding and to potentiate or inhibit [3H]MK-801 binding. All quinoxalines tested were invariably more potent displacers of [3H]DCKA binding than [3H]Gly binding, whereas kynurenines were similarly effective in displacing the binding of both [3H]Gly and [3H]DCKA. These results undoubtedly give support to the proposal that [3H]DCKA is one useful radioligand available in terms of its high selectivity and affinity for the Gly domain in the brain. Possible multiplicity of the Gly domain is suggested by the differential pharmacological profiles between the binding of [3H]Gly and [3H]DCKA.

Animals↗
PubMed1993

Further evidence for multiple forms of an N-methyl-D-aspartate recognition domain in rat brain using membrane binding techniques.

Pretreatment with sulfhydryl-reactive agents, such as N-ethylmaleimide and p-chloromercuriphenylsulfonic acid, invariably resulted in marked inhibition of the binding of DL-(E)-2-amino-4-[3H]propyl-5-phosphono-3-pentenoic acid ([3H]CGP 39653), a competitive antagonist at an N-methyl-D-aspartate (NMDA)-sensitive subclass of central excitatory amino acid receptors, in brain synaptic membranes extensively washed and treated with Triton X-100, but did not significantly affect the binding of L-[3H]-glutamic acid ([3H]Glu), an endogenous agonist. The pretreatment was effective in reducing the binding of [3H]-CGP 39653 at equilibrium, without altering the initial association rate, and decreased the affinity for the ligand. Pretreatment with sulfhydryl-reactive agents also enhanced the potencies of NMDA agonists to displace [3H]-CGP 39653 binding and attenuated those of NMDA antagonists, but had little effect on the potencies of the agonists and antagonists to displace [3H]Glu binding. The binding of both [3H]CGP 39653 and [3H]Glu was similarly sensitive to pretreatment with four different proteases in Triton-treated membranes, whereas pretreatment with phospholipase A2 or C markedly inhibited [3H]CGP 39653 binding without altering [3H]Glu binding. Moreover, both phospholipases not only induced enhancement of the abilities of NMDA agonists to displace the binding of [3H]CGP 39653 and [3H]Glu, but also caused diminution of those of NMDA antagonists. These results suggest that both sulfhydryl-reactive agents and phospholipases may predominantly interfere with radiolabeling of the NMDA recognition domain in a state favorable to an antagonist by [3H]CGP 39653, with concomitant facilitation of that in an antagonist-preferring form by [3H]Glu. The possible presence of multiple forms of the NMDA recognition domain is further supported by these data.

2-Amino-5-phosphonovalerate↗
PubMed1993

[Higher production of tumor necrosis factor (TNF) elicited by a biological response modifier (BRM) in aging mice].

We investigated age-related changes in the capacity for tumor necrosis factor (TNF) production in young and aged inbred C3H/He mice by injecting them with OK-432, a biological response modifier (BRM). An intravenous injection of 0.4mg of OK-432 was found to induce TNF production and two consecutive injections of 2KE of OK-432 induced much higher TNF production. Both the single and two consecutive injections of OK-432 induced significantly higher TNF production in aged mice than in young ones. Furthermore, the TNF-productive response to the two consecutive injections of OK-432 seemed to increase with aging. Male mice tended to show a marginally higher TNF-productive response than females. The mechanism by which aged mice have a higher capacity for TNF production is not clear. The following possibilities are conceivable. 1) Macrophages which are major TNF producer cells may be activated in aged mice. 2) Specific T cells which are cross-reactive to antigenic determinants in OK-432 may be increased in number in aging mice and activate macrophages effectively to produce TNF when stimulated by OK-432. In general, immunological functions tend to decline with aging. Our present results, however, suggest that by using an appropriate BRM we may be able to induce higher TNF production in the aged. This might lead to effective prevention and therapy for tumors, which increase in incidence with age.

Aging↗
PubMed1993

Novel preservation solution permits 24-hour preservation in rat and baboon cardiac transplant models.

BACKGROUND: Cardiac preservation for transplantation disrupts normal vascular homeostatic mechanisms. Hypoxia and reoxygenation increase endothelial cell permeability, induce procoagulant activity, and alter endothelial cell/leukocyte interactions, with a parallel reduction in endothelial cAMP and nitric oxide levels. Because hypoxia/reoxygenation simulates a significant component of the global ischemia/reperfusion of cardiac transplantation, we hypothesized that preservation failure may be related to these perturbations. This work focuses on repleting the intercellular/intracellular second messengers nitric oxide/cGMP and cAMP in the donor heart to enhance cardiac preservation for transplantation. METHODS AND RESULTS: A heterotopic rat heart transplant model was used to compare lactated Ringer's (LR) and University of Wisconsin (UW) solutions to a novel storage solution (Columbia University solution, CU), which contains a cAMP analogue (dibutyryl cAMP) and nitroglycerin (to enhance nitric oxide-related mechanisms). By 24 to 28 hours of preservation, no LR hearts survived (n = 9), 35% of UW hearts survived (n = 20), and 88% of CU-preserved hearts survived (n = 8; P < .05) versus LR or UW). The viability of preserved hearts was explored by determining whether CU preservation enabled myocytes to maintain resting membrane potentials and preserve their ability to generate an action potential in response to a field stimulus. Of 24 sites explored with a microelectrode in UW-preserved hearts, only 4% were able to generate an action potential, compared with 75% of 36 sites in CU-preserved hearts (P < .001), with corresponding preservation of resting membrane potential in the CU-preserved hearts (-13 mV for UW versus -54 mV for CU, P < .001). Orthotopic baboon cardiac transplantation performed after 24-hour simple hypothermic preservation demonstrated that no UW-preserved heart (n = 4) survived the perioperative period; in contrast, four of five hearts stored for 24 hours in CU solution sustained the recipient with minimal inotropic support, and two animals survived long-term. CONCLUSIONS: Sustaining higher levels of intercellular/intracellular second messengers cAMP and nitric oxide/cGMP provides a new approach to enhancing cardiac preservation.

Adenosine↗
PubMed1992

Differential modulation by divalent cations of [3H]MK-801 binding in brain synaptic membranes.

Endogenous divalent cations, such as Mg2+, Ca2+, and Zn2+, differentially affected the binding of (+)-[3H]5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imi ne maleate ([3H]MK-801) to an ion channel associated with an N-methyl-D-aspartate-sensitive subclass of excitatory amino acid receptors in different preparations of brain synaptic membranes. Both Mg2+ and Ca2+ were weak inhibitors of the binding in membranes which had not been extensively washed (nonwashed membranes), over a concentration range effective in markedly potentiating the binding in the absence of any added stimulants in membranes which had been extensively washed, but not treated with a detergent (untreated membranes). In membranes extensively washed and treated with Triton X-100 (Triton-treated membranes), both cations significantly potentiated the binding in the presence of added glutamate alone. In contrast, Zn2+ was invariably active as a potent inhibitor of the binding irrespective of the membrane preparations used. In untreated membranes, Ca2+ markedly accelerated the initial association rate of [3H]MK-801 binding without affecting the binding at equilibrium in a manner similar to that found with glycine, as well as with glutamate; Mg2+, however, facilitated the initial association rate with a concomitant reduction of the binding at equilibrium. Zn2+ was effective in accelerating the initial rapid phase of association, with the initial slow phase being delayed, and in markedly reducing the binding at equilibrium. Both Mg2+ and Ca2+ also facilitated dissociation of the bound [3H]MK-801 and Zn2+ slowed the dissociation in untreated membranes.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

[Mechanisms of protective action of radix Salviae miltiorrhizae (RSM) against experimental hepatic injury in rats].

The results indicated that RSM could significantly inhibit the lipid peroxidation of normal livers and cultured hepatocytes of rats, induce liver microsomal cytochrome P450 in normal rats, increase nucleic acids, proteins, urea and cerolloplasmin of damaged cultured hepatocytes of rats, relieve ultrastructural damage of cultured hepatocytes induced by CCl4. The pharmacological actions mentioned above should be considered as important mechanisms of RSM against liver injury.

Animals↗
PubMed1992

[Evaluation of compound action potential tuning curves from normal guinea pig cochleas].

Eighteen ears of normal guinea pigs were examined by means of compound action potential tuning curve (AP-TC) with different measure parameters. AP-TC was measured at pure-tone of various frequencies using simultaneous masking procedure by the electrodes on the round window. AP was evoked by filter click (2, 3, 4, 5, 6, 10 kHz) with an intensity 10, 30 and 50 dB above threshold. Masker level producing 30% reduction in AP amplitude was used. The results showed that AP-TC was more sharply as test frequency was elevated. It is corresponded well with the published results. No differences in tuning (i.e., the form of the curves and Q10 dB values) and thresholds were found between two different times for the same animal. In addition, the Q10 dB values of AP-TC were gradually reduced along with the elevate of the test stimuli intensity from 10 to 50 dB above threshold.

Action Potentials↗
PubMed1991

Free radical recycling and intramembrane mobility in the antioxidant properties of alpha-tocopherol and alpha-tocotrienol.

d-Alpha-tocopherol (2R,4'R,8'R-Alpha-tocopherol) and d-alpha-tocotrienol are two vitamin E constituents having the same aromatic chromanol "head" but differing in their hydrocarbon "tail": tocopherol with a saturated and toctrienol with an unsaturated isoprenoid chain. d-Alpha-tocopherol has the highest vitamin E activity, while d-alpha-tocotrienol manifests only about 30% of this activity. Since vitamin E is considered to be physiologically the most important lipid-soluble chain-breaking antioxidant of membranes, we studied alpha-tocotrienol as compared to alpha-tocopherol under conditions which are important for their antioxidant function. d-Alpha-tocotrienol possesses 40-60 times higher antioxidant activity against (Fe2+ + ascorbate)- and (Fe2+ + NADPH)-induced lipid peroxidation in rat liver microsomal membranes and 6.5 times better protection of cytochrome P-450 against oxidative damage than d-alpha-tocopherol. To clarify the mechanisms responsible for the much higher antioxidant potency of d-alpha-tocotrienol compared to d-alpha-tocopherol, ESR studies were performed of recycling efficiency of the chromanols from their chromanoxyl radicals. 1H-NMR measurements of lipid molecular mobility in liposomes containing chromanols, and fluorescence measurements which reveal the uniformity of distribution (clusterizations) of chromanols in the lipid bilayer. From the results, we concluded that this higher antioxidant potency of d-alpha-tocotrienol is due to the combined effects of three properties exhibited by d-alpha-tocotrienol as compared to d-alpha-tocopherol: (i) its higher recycling efficiency from chromanoxyl radicals, (ii) its more uniform distribution in membrane bilayer, and (iii) its stronger disordering of membrane lipids which makes interaction of chromanols with lipid radicals more efficient. The data presented show that there is a considerable discrepancy between the relative in vitro antioxidant activity of d-alpha-tocopherol and d-alpha-tocotrienol with the conventional bioassays of their vitamin activity.

Animals↗
PubMed1991

Simplified pressure method for respirator fit testing.

A simplified pressure method has been developed for fit testing air-purifying respirators. In this method, the air-purifying cartridges are replaced by a pressure-sensing attachment and a valve. While wearers hold their breath, a small pump extracts air from the respirator cavity until a steady-state pressure is reached in 1 to 2 sec. The flow rate through the face seal leak is a unique function of this pressure, which is determined once for all respirators, regardless of the respirator's cavity volume or deformation because of pliability. The contaminant concentration inside the respirator depends on the degree of dilution by the flow through the cartridges. The cartridge flow varies among different brands and is measured once for each brand. The ratio of cartridge to leakflow is a measure of fit. This flow ratio has been measured on human subjects and has been compared to fit factors determined on the same subjects by means of photometric and particle count tests. The aerosol tests gave higher values of fit.

Humans↗
PubMed1991

Respirator fit and protection through determination of air and particle leakage.

A laboratory technique for determining the respirator protection factor from a test of fit is described. A dynamic pressure test quantifies the air flow through the leak. Calibration data, stored in a computer, relate the contaminant influx to this air flow, and a similar pressure test determines the flow through the respirator cartridges and, therefore, the dilution characteristics. Contaminant removal characteristics of the cartridges are stored in the computer. The contaminant penetration is calculated from these data on flow and removal efficiency. Through specification of the aerosol size distribution and the method of measurement, protection factors are calculated for specific work environments, work loads and respirator cartridges. The protection factor is shown to be highly dependent on the method of measuring the contaminant and on the cartridges used.

Air Pollutants↗
PubMed1991

[Intrauterine adhesion and ovulatory disorder following induced abortion].

Follicular stimulating hormone, luteinizing hormone, prolactin, estradiol and progesterone were measured by radioimmunoassay in 34 patients with those from 34 normal cycling women after induced abortion. The result showed that the anovulatory incidence of the patients (20.6%) was significantly higher than that of the controls (2.8%) (P less than 0.05). Increased prolactin and premature ovarian failure might be responsible for the anovulatory disorder, resulting in unsatisfactory restoration of fertility. It is suggested that the low success rate in treating patients with intrauterine adhesion after induced abortion may be due to premature ovarian failure and hyperprolactinaemia.

Abortion, Induced↗
PubMed1991

[Relation between changes in compound action potential tuning curves and the pathology of cochlear hair cells stereocilia].

Relations of the changes of compound action potential tuning curve (AP-TC) and AP(N1) threshold to the pathology of the cochlear hair cells stereocilia were examined in acoustic-traumatized guinea pigs. The results showed that all animals with hair cell stereocilia damaged showed deteriorations of AP-TC. It supports the suggestion that mechanical properties of hair cell stereocilia contribute to the frequency selectivity of cochlea. After exposure, the characteristic frequency (CF) shift of AP-TC was observed. The mechanism of CF shift may be related to the lesion of hair cell stereocilia which may change normal tono-topic on cochlear level. Correlation between the damages of inner/outer hair cell stereocilia and changes of AP-TC and AP threshold suggested that the threshold and tuning properties of the cochlea are dependent upon the integrity of the inner and outer hair cells and their inter-relationship.

Action Potentials↗