Biomedical subjects

Q X Chen

Publications and source records attributed to Q X Chen.

At least 37 records · Page 2Linked to original sources

PubMed1998

An essential lysine residue of green crab (Scylla Serrata) alkaline phosphatase.

The values of pKa (10.38) and Hion (10.92 Kcal/mol) have been determined for the ionizing groups controlling activity of green crab alkaline phosphatase. The results suggest that -NH2 of lysine residue responsible for the ionization with pKc = 10.38 and delta H(o)ion = 10.92 Kcal/mol is in the active site of the enzyme. Modification of lysine residues of the enzyme by an excess of 2,4,6-trinitrobenzenesulfonic acid leads to complete inactivation. The two results coincide with each other. Quantitative assessment of the data indicates that among the reactive -NH2 groups modified only one is essential for the activity of the enzyme.

Alkaline Phosphatase↗

PubMed1998

Zn2+ blocks the NMDA- and Ca2+ -triggered postexposure current ipe in hippocampal pyramidal cells.

Whole cell voltage-clamp recordings from acutely isolated hippocampal CA1 pyramidal cells from adult guinea pigs were used to evaluate divalent cations as possible blockers of the postexposure current (Ipe). Ipe is a cation current that is triggered by the rise in intracellular Ca2+ concentration that occurs after the application of a toxic level of N-methyl-D-aspartate (NMDA). Once triggered, Ipe continues to grow until death of the neuron occurs. Ipe may be a critical link between transient NMDA exposure and cell death. Ipe was blocked by micromolar concentrations of Zn2+. The Zn2+ effect had an IC50 of 64 microM and saturated at 500 microM. Prolonged Zn2+ block of Ipe revealed that the maintenance of a steady Ipe is not dependent on Ipe-mediated Ca2+ influx but that the continuous growth in Ipe is dependent on Ipe-mediated Ca2+ influx. The availability of an effective blocker of Ipe should facilitate the investigation of the intracellular activation pathway of Ipe and the role of Ipe in neuronal death.

Action Potentials↗

PubMed1998

Kinetics of inactivation of Penaeus penicillatus acid phosphatase during inhibition by N-bromosuccinimide.

In the present investigation, the inactivation by N-bromosuccinimide of acid phosphatase from penaeus penicillatus has been studied using the kinetic method of the substrate reaction during modification of enzyme activity as previously described by Tsou [(1988, Adv. Enzymemol. Related Areas Mol. Biol. 61, 381-436]. The results show that inactivation of the enzyme by N-bromosuccinimide is a slow, reversible reaction. The results also clearly show that the modification of the tryptophan residues of penaeus penicillatus acid phosphatase by high concentrations of N-bromosuccinimide led to the complete inactivation of the enzyme. The microscopic rate constants were determined for the reaction of the inactivator with the free enzyme and with the enzyme-substrate complex. Comparison of the obtained microscopic rate constants indicates that the presence of the substrate offers marked protection of the enzyme against inactivation by N-bromosuccinimide. The above results suggest that the tryptophan residue is essential for activity and may be situated at the active site of the enzyme.

Acid Phosphatase↗

PubMed1997

Secondary activation of a cation conductance is responsible for NMDA toxicity in acutely isolated hippocampal neurons.

One of the key questions concerning glutamate toxicity is how a transient NMDA exposure can lead to a delayed death of neurons. To address this issue, we performed whole-cell recording on acutely isolated hippocampal CA1 neurons to monitor the membrane response after NMDA exposure. Transient NMDA exposure (100 microM, 10 min) induced an inward current (postexposure current; Ipe) which was associated with a Ca2+- and Na+-permeable cation conductance. Ipe continuously increased (in the absence of NMDA) until death of the neuron occurred. Application of NMDA in the absence of extracellular calcium failed to trigger Ipe and neuronal death. Postexposure suppression of Ipe protected against NMDA toxicity. These results indicate that a cation current, which is induced by an increase in intracellular calcium concentration ([Ca2+]i) and is itself partly carried by Ca2+, links the initial NMDA exposure to neuronal death.

Animals↗

PubMed1997

An essential tryptophan residue of green crab (syclla serrata) alkaline phosphatase.

The tryptophan residues in green crab (scylla serrata) alkaline phosphatase (EC 3.1.3.1) have been modified by N-bromosuccinimide (NBS). The modification of five tryptophan residues leads to complete loss of enzymatic activity. With the increase of NBS concentration, both the absorption at 278 nm and the fluorescence emission intensity at 335 nm of the modified enzyme decreased markedly indicating the modification of tryptophan residues. Quantitative treatment of the data (Tsou, Sci. Sinica 1962, 11, 1535-1558) shows that among the tryptophan residues modified, one is essential for its catalytic activity. The presence of the substrate markedly protects the modification of tryptophan residues as well as the inactivation, suggesting that the essential tryptophan residue is situated at the active site of this enzyme.

Alkaline Phosphatase↗

PubMed1997

Unfolding and inactivation of Penaeus penicillatus acid phosphatase during denaturation by guanidine hydrochloride.

The conformational changes of penaeus penicillatus acid phosphatase during denaturation in guanidine hydrochloride solutions were studied by following changes in the intrinsic fluorescence, ultraviolet difference absorption, and circular dichroism spectra. Inactivation of the enzyme in guanidine hydrochloride solutions were compared with unfolding of the enzyme molecule. The results show that the extent of unfolding in guanidine solutions measured by several different methods closely coincide with each other and that slightly lower concentrations of guanidine are required to bring about inactivation than are required to produce significant conformational changes of the enzyme molecule. At the same concentrations, the inactivation rate constants are markedly faster than the rate constants for unfolding of the enzyme. The above results suggest that the active sites of this enzyme display more conformational flexibility than the enzyme molecule as a whole.

Acid Phosphatase↗

PubMed1997

Kinetics of the thermal inactivation of alkaline phosphatase from green crab (Scylla serrata).

The kinetics of thermal inactivation of alkaline phosphatase from green crab (Scylla Serrata) has been studied using the kinetic method relating to the substrate reaction during irreversible inhibition of enzyme activity previously described by Tsou. The results show that the thermal inactivation of the enzyme is an irreversible reaction. Comparison of the microscopic rate constants for thermal inactivation of free enzyme and the enzyme-substrate complex shows that the presence of substrate has a certain protective effect against thermal inactivation.

Alkaline Phosphatase↗

PubMed1996

Kinetics of inhibition of alkaline phosphatase from green crab (Scylla serrata) by N-bromosuccinimide.

The inactivation of alkaline phosphatase from green crab (Scylla serrata) by N-bromosuccinimide has been studied using the kinetic method of the substrate reaction during modification of enzyme activity previously described by Tsou [(1988), Adv. Enzymol. Related Areas Mol. Biol. 61, 381-436]. The results show that inactivation of the enzyme is a slow, reversible reaction. The microscopic rate constants for the reaction of the inactivator with free enzyme and the enzyme-substrate complex were determined. Comparison of these rate constants indicates that the presence of substrate offers marked protection of this enzyme against inactivation by N-bromosuccinimide. The above results suggest that the tryptophan residue is essential for activity and is situated at the active site of the enzyme.

Alkaline Phosphatase↗

PubMed1996

Comparison of inactivation and unfolding of green crab (Scylla serrata) alkaline phosphatase during denaturation by guanidinium chloride.

Green crab (Scylla serrata) alkaline phosphatase (EC 3.1.3.1) is a metalloenzyme, each active site in which contains a tight cluster of two zinc ions and one magnesium ion. Unfolding and inactivation of the enzyme during denaturation in guanidinium chloride (GuHCl) solutions of different concentrations have been compared. The kinetic theory of the substrate reaction during irreversible inhibition of enzyme activity previously described by Tsou [(1988), Adv. Enzymol. Related Areas Mol. Biol. 61, 381-436] has been applied to a study on the kinetics of the course of inactivation of the enzyme during denaturation by GuHCl. The rate constants of unfolding and inactivation have been determined. The results show that inactivation occurs before noticeable conformational change can be detected. It is suggested that the active site of green crab alkaline phosphatase containing multiple metal ions is also situated in a limited region of the enzyme molecule that is more fragile to denaturants than the protein as a whole.

Alkaline Phosphatase↗

PubMed1996

Kinetics of inactivation of green crab (Scylla Serrata) alkaline phosphatase during removal of zinc ions by ethylenediaminetetraacetic acid disodium.

Green crab (Scylla Serrata) alkaline phosphatase (EC 3.1.3.1) is a metalloenzyme, the each active site in which contains a tight cluster of two zinc ions and one magnesium ion. The kinetic theory of the substrate reaction during irreversible inhibition of enzyme activity previously described by Tsou has been applied to a study on the kinetics of the course of inactivation of the enzyme by ethylenediaminetetraacetic acid disodium (EDTA). The kinetics of the substrate reaction with different concentrations of the substrate p-nitrophenyl phosphate (PNPP) and inactivator EDTA suggested a complexing mechanism for inactivation by, and substrate competition with, EDTA at the active site. The inactivation kinetics are single phasic, showing the initial formation of an enzyme-EDTA complex is a relatively rapid reaction, followed a slow inactivation step that probably involves a conformational change of the enzyme. Zinc ions are finally removed from the enzyme. The presence of metal ions apparently stabilizes an active-site conformation required for enzyme activity.

Alkaline Phosphatase↗

PubMed1996

Kinetics of inhibition of green crab (Scylla serrata) alkaline phosphatase by phenylglyoxal.

The kinetics method of the substrate reaction during modification of enzyme activity previously described by Tsou (Adv. Enzymol. Related. Areas Mol. Biol., 1988, 61, 381-436) has been applied to study the kinetics of the course of inactivation of green crab alkaline phosphatase by phenylglyoxal. The obtained results shows that the inactivation of the enzyme by phenylglyoxal is a slow reversible reaction. The microscopic rate constants for the reaction of the inhibitor with the enzyme were determined. The presence of substrate offers marked protection of this enzyme against inactivation by phenylglyoxal. The above results suggest that the arginine residue is essential for activity and is situated at the active site of the enzyme.

Alkaline Phosphatase↗

PubMed1996

[3.3-style nurses' training goal made by the ABC analytical method].

Statistics of the structure of educational level and worked years of 140 occupied nurses, analysed by ABC curvegram, showed that the key training objects are class A (1 to 10 years at work) and class B nurses. Then the goal of nurses' training at work during 3 years for class A and B was made according to the difference of their classes and need of knowledge. In order to improve the effect of 3.3 style nurse training goal a method of two-direction teaching and assessing were applied, actual effect was achieved during a three-year conduction.

China↗

PubMed1995

Suppression of GABAA receptor responses by NMDA application in hippocampal neurones acutely isolated from the adult guinea-pig.

1. In acutely isolated hippocampal cells, NMDA and glutamate application suppressed GABAA receptor-mediated responses. We studied the cellular events underlying the interaction between the two classes of receptors by using a whole-cell voltage-clamp approach. 2. Following an NMDA application, an outward current mediated by GABAA receptor activation (GABA response) was suppressed for up to 12 s. The suppression of the GABA response was reduced when Ca2+ in the extracellular solution was replaced by Ba2+ or when intracellular BAPTA (1,2-bis(O-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid) was increased from 1 to 10 mM. 3. Replacing ATP in the intracellular solution by adenosine-5'-O-3-thiotriphosphate reduced the suppressive effect of NMDA application on the GABA response. Okadaic acid, a phosphatase inhibitor, also prevented the NMDA-induced suppression of the GABA response. In addition, when the intracellular perfusing solution contained the calcineurin autoinhibitory fragment (50 microM), suppression of the GABA response by the NMDA current was also reduced. 4. Intracellular perfusion of an activated form of the Ca(2+)-dependent phosphatase, calcineurin, suppressed GABA responses. 5. The results show that NMDA responses elicited in hippocampal neurones transiently suppressed GABA responses. The data suggest that the functional linkage of the NMDA response with the GABA response was established via a Ca(2+)-dependent dephosphorylation process.

Animals↗

PubMed1995

Suppression of a Ca2+ current by NMDA and intracellular Ca2+ in acutely isolated hippocampal neurons.

1. Ca2+ current was examined in acutely isolated hippocampal cells with the use of whole cell voltage-clamp recording and under continuous intracellular perfusion. A persistent Ca2+ current was activated by depolarization to -10 mV from a holding potential of -50 mV. 2. The persistent Ca2+ current was suppressed upon a wash out of the intracellular Mg(2+)-ATP. Adenosine 3',5'-cyclic monophosphate (cAMP) introduced intracellularly potentiated the Ca2+ current, and kinase A inhibitor blocked the current. 3. Reversible suppression of the persistent Ca2+ current was also observed by elevating intracellular Ca2+. This Ca(2+)-dependent suppression was retarded by the addition of a phosphatase inhibitor, okadaic acid, to the intracellular solution. 4. N-methyl-D-aspartate (NMDA) elicited inward current (NMDA response) in the isolated cells. The persistent Ca2+ current was transiently suppressed after the NMDA response. Suppression of the Ca2+ current by NMDA was reduced when intracellular Ca2+ buffering capacity was increased by increasing the concentration of bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA) from a concentration of 1-10 mM. 5. Substitution of ATP in the intracellular solution with ATP-gamma-S or the addition of okadaic acid to the intracellular solution reduced the suppressive effect of NMDA on the Ca2+ current. 6. The results suggest that the persistent Ca2+ current in the hippocampal cells is maintained by a kinase A-mediated phosphorylation. Increases in the intracellular Ca2+ concentration suppressed the Ca2+ current via a mechanism involving a phosphatase. Ca2+ entry through the NMDA receptor channel suppressed the Ca2+ channel by activating the phosphatase.

Adenosine Triphosphate↗

PubMed1994

A new laser-ultrasound transducer for medical applications.

A new type of probe is presented, which combines an ultrasound receiver constructed from piezoelectric polymer materials with a coaxial optical fibre for laser-induced ultrasound. The initial application of this probe, which may be further miniaturized, is in intra-arterial imaging and therapy. Overall diameters of the prototype probes presently range from 2.5 mm to 10.0 mm. Pulsed laser beams are introduced through the optical fibre to form a photoacoustic source within the material under investigation. Ultrasonic waves are induced in the material due to the photo-acoustic effect and received on return by a forward-looking piezoelectric polymer transducer. In principle, other high-power laser beams can also be introduced through the same optical fibre for therapeutic use. Initial development and characterization of this combined probe is reported in this paper, together with some results.

Acoustics↗

PubMed1994

Analysis of tuberculous meningitis cases by an immunoblotting assay based on a mycobacterial antigen complex.

Tuberculous meningitis cases were analyzed by an immunoblotting test based on Mycobacterium bovis BCG antigen complex A60. Anti-A60 immunoglobulin G (IgG) in cerebrospinal fluid (CSF) allowed early diagnosis, and concentrations decreased after recovery. In primary meningitis forms, anti-A60 IgGs were intrathecally synthesized and specific oligoclonal IgGs were present in CSF. In meningeal complications of pulmonary tuberculosis, there were matching titers of anti-A60 IgG in blood and CSF (mirror pattern). Correlation between CSF-restricted patterns and CSF pleocytosis was shown.

Adolescent↗

PubMed1994

Intracellular control of GABAA-receptor responses in hippocampal neurons: role of phosphorylation.

Recent studies show that GABAA-receptors mediated responses in the hippocampal cells are suppressed upon increases in intracellular Ca2+ [Ca2+]i. A series of experiments were carried out to evaluate the physiological significance of this modulatory action of [Ca2+]i. We show that GABAA-responses are suppressed by a preceding NMDA application in hippocampal cells. Furthermore the suppressive effect is mediated via an increase in [Ca2+]i. Additional data reveal that NMDA application suppressed the GABAA response by initiating an Ca2+ dependent dephosphorylation process and the suppression is prevented by intracellular ATP-gamma-S or okadaic acid. Our study suggest that there interactions between the excitatory synaptic responses and the inhibitory synaptic responses may be effected with intracellular Ca2+ serving as a second messenger.

Animals↗

PubMed1992

Different effects of intracellular and extracellular TEA on voltage-dependent K currents of acutely isolated hippocampal CA1 neurons.

Tetraethylammonium (TEA) effects on K currents were examined on either side of the membrane of hippocampal CA1 neurons by means of whole-cell voltage-clamp recording and intracellular perfusion. Recording media contained ion channel blockers to allow the selective activation of voltage-dependent K currents which consisted of a rapidly decaying component (A-current) and a delayed component. Voltage protocols were applied to separate the A-current from the delayed component. Results show that 10 mM extracellular TEA suppressed 50 +/- 11% (S.D., n = 4) of the delayed current at different levels of depolarization but had little effect on the A-current. In contrast 10 mM TEA applied by intracellular perfusion suppressed the A-current by 42 +/- 10% (S.D., n = 4) in addition to inhibiting the delayed currently 55 +/- 15% (S.D., n = 4). Both the intracellular and extracellular actions of TEA on K currents showed no voltage- nor time-dependency. The results suggest that voltage-dependent transient current (A-current) is mediated through a separate group of ionic channels distinct from those that sustained the delayed current. Furthermore, the asymmetrical effects of intracellular and extracellular TEA on the transient current are similar to those described for the A-current in molluscan neurons. This observation supports the notion that the structure of the ion channel mediating the A-current is closely conserved across different species.

Animals↗