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Biomedical subjects

M C Chung

Publications and source records attributed to M C Chung.

At least 109 records · Page 6Linked to original sources
PubMed1992

Purification and characterization of a novel enzyme, arylalkyl acylamidase, from Pseudomonas putida Sc2.

A novel enzyme, arylalkyl acylamidase, which shows a strict specificity for N-acetyl arylalkylamines, but not acetanilide derivatives, was purified from the culture broth of Pseudomonas putida Sc2. The purified enzyme appeared to be homogeneous, as judged by native and SDS/PAGE. The enzyme has a molecular mass of approximately 150 kDa and consists of four identical subunits. The purified enzyme catalyzed the hydrolysis of N-acetyl-2-phenylethylamine to 2-phenylethylamine and acetic acid at the rate of 6.25 mumol.min-1.mg-1 at 30 degrees C. It also catalyzed the hydrolysis of various N-acetyl arylalkylamines containing a benzene or indole ring, and acetic acid arylalkyl esters. The enzyme did not hydrolyze acetanilide, N-acetyl aliphatic amines, N-acetyl amino acids, N-acetyl amino sugars or acylthiocholine. The apparent Km for N-acetylbenzylamine, N-acetyl-2-phenylethylamine and N-acetyl-3-phenylpropylamine are 41 mM, 0.31 mM and 1.6 mM, respectively. The purified enzyme was sensitive to thiol reagents such as Ag2SO4, HgCl2 and p-chloromercuribenzoic acid, and its activity was enhanced by divalent metal ions such as Zn2+, Mg2+ and Mn2+.

Amidohydrolases↗
PubMed1992

Brain glutamine synthetase increases following cerebral ischemia in the rat.

Changes in astrocyte glutamine synthetase (GS) in postischemic rat brain were evaluated and correlated with regional neuronal vulnerability or resistance to ischemia. Rats subjected to 20 or 30 min of cerebral ischemia were allowed to survive for 3 or 24 h after ischemia; normal animals served as controls. Resultant neuronal necrosis was severe in the striatum by 24 h and in the CA1 region of the hippocampus at 72 h; neurons in paramedian cortex and CA3 region of the hippocampus were not permanently damaged. Glutamine synthetase (GS) immunocytochemistry was performed on vibratome sections of paraformaldehyde-fixed brains and enzyme activity was assayed in frozen samples of cerebral cortex, striatum and hippocampus. At 3 and 24 h after ischemia, GS immunoreactivity increased and was secondary to enlargement of GS-positive cell bodies and processes as well as to increased numbers of GS-positive astrocytes. Enzyme activity also increased in cortex, striatum and hippocampus at 3 and 24 h (P less than or equal to 0.03). This study shows that increase in astrocyte GS occurs rapidly after ischemia, and prior studies indicate that this increase occurs in parallel with proliferative changes in astrocyte organelles. The results also suggest that astrocyte metabolism of glutamate increases after ischemia. The increased capacity for glutamine synthetase may be important in normalizing extracellular glutamate following ischemia and protecting brain from the neurotoxic effects of this excitatory amino acid.

Animals↗
PubMed1991

Localization of specific repetitive DNA sequences in individual rice chromosomes.

This paper describes the characterization and chromosomal distribution of three different rice (Oryza sativa) repetitive DNA sequences. The three sequences were characterized by sequence analysis, which gave 355, 498 and 756 bp for the length of the repeat unit in Os48, OsG3-498 and OsG5-756, respectively. Copy number determination by quantitative DNA slot-blot hybridization analysis showed 4000, 1080 and 920 copies, respectively, per haploid rice genome for the three sequences. In situ DNA hybridization analysis revealed that 95% of the silver grains detected with the Os48 probe were localized to euchromatic ends of seven long arms and one short arm out of the 12 rice chromosomes. For the OsG3-498 repetitive sequence, the majority of silver grains (58%) were also clustered at the same chromosomal ends as that of Os48. The minority (28%) of silver grains were located at heterochromatic short arms and centromeric regions. For the OsG5-756 repetitive sequence, 81% of the silver grains labeled the heterochromatic short arms and regions flanking all of the 12 centromeres. Thus, each of these three repetitive sequences was distributed at specific defined chromosomal locations rather than randomly at many chromosomal locations. The approximate copy number of a given repetitive DNA sequence at any specific chromosomal location was calculated by combining the information from in situ DNA hybridization analysis and the total copy number as determined by DNA slot-blot hybridization.

Base Sequence↗
PubMed1991

Purification of transferrins and lactoferrin using DEAE affi-gel blue.

1. A simple method for purifying transferrins and lactoferrin is described. 2. The method consists of a preliminary step of dye-ligand chromatography using DEAE Affi-Gel Blue as the gel matrix at pH 7.5. In this chromatographic step, the transferrins and lactoferrin were readily separated from the bulk of the other proteins by start buffer elution. 3. Differences in the chromatographic behaviour of the various serum transferrins (monkey, human, rabbit, pig, chicken and duck) and ovotransferrin upon DEAE Affi-Gel Blue chromatography can be attributed to differences in the anionic charge of the transferrins in 0.02 M potassium phosphate buffer, pH 7.5, thus resulting in the differential retardation of these protein molecules by the gel matrix. 4. The result of DEAE Affi-Gel Blue chromatography of human lactoferrin is different from that for the transferrins. This may possibly reflect the differences in the strength of interaction between lactoferrin and transferrin with this gel matrix.

Amino Acid Sequence↗
PubMed1988

Ketopantoic acid reductase of Pseudomonas maltophilia 845. Purification, characterization, and role in pantothenate biosynthesis.

Ketopantoic acid reductase (EC 1.1.1.169), an enzyme that catalyzes the formation of D-(-)-pantoic acid from ketopantoic acid, was purified 6,000-fold to apparent homogeneity with a 35% overall recovery from Pseudomonas maltophilia 845 and then crystallized. The relative molecular mass of the native enzyme, as estimated by the sedimentation equilibrium method, is 87,000 +/- 5,000, and the subunit molecular mass is 30,500. The enzyme shows high specificity for ketopantoic acid as a substrate (Km = 400 microM, Vm = 1,310 units/mg of protein) and NADPH as a coenzyme (Km = 31.8 microM). Only 2-keto-3-hydroxyisovalerate (Km = 8.55 mM, Vm = 35.8 units/mg) was reduced among a variety of other carbonyl compounds tested. The reaction is reversible (Km for D-(-)-pantoic acid = 52.1 mM), although the reaction equilibrium greatly favors the direction of D-(-)-pantoic acid formation. That the enzyme is responsible for the synthesis of D-(-)-pantoic acid necessary for the biosynthesis of pantothenic acid in P. maltophilia 845 is indicated by the observations that only this enzyme is missing in D-(-)-pantoate (or pantothenate)-requiring mutants derived from P. maltophilia 845 among several enzymes (i.e. ketopantoyl lactone reductase (EC 1.1.1.168) and acetohydroxy acid isomeroreductase (EC 1.1.1.86], which may be concerned in the formation of D-(-)-pantoic acid, assayed, whereas it is present in substantial amounts in the parent strain and in spontaneous revertants of the mutants.

Alcohol Oxidoreductases↗
PubMed1986

Acute and chronic effects of haloperidol on plasma and brain homovanillic acid in the rat.

The effects of acute and chronic administration of haloperidol on homovanillic acid (HVA) in plasma and the brain were examined in the rat. Acute haloperidol treatment (1 mg/kg) resulted in highly significant elevations in HVA within 30 min and produced a maximal increase of HVA in 3-6 hr in both plasma and the whole brain. The response of brain HVA to increasing doses (0.05-30 mg/kg) of haloperidol showed an inverted U pattern. Plasma HVA showed a very flat response to lower doses (less than or equal to 5 mg/kg) of haloperidol and a dramatically elevated one to higher doses (greater than or equal to 10 mg/kg). Haloperidol produced a parallel increase in plasma and brain HVA at lower doses (less than or equal to 2 mg/kg) only. After chronic administration of haloperidol for up to 28 days, the response of HVA in plasma correlated mainly with, but tolerated later than, those in the whole brain and the olfactory tubercle.

Animals↗
PubMed1985

A specific iron stain for iron-binding proteins in polyacrylamide gels: application to transferrin and lactoferrin.

A new method for specifically staining the iron atoms present in transferrin and lactoferrin after polyacrylamide gel electrophoresis and isoelectric focusing is described. The stain, 3-(2-pyridyl)-5,6-bis(2-(5-furylsulfonic acid))-1,2,4-triazine, disodium salt, or Ferene S, will detect transferrin in 5 microliters of human serum, lactoferrin in 10 microliters of human whey, and 10 micrograms of purified primate (Macaca fascicularis) transferrin. This method of staining is very rapid as the serum transferrin bands can be seen within 5 to 10 min of staining.

Animals↗
PubMed1985

Primate (Macaca fascicularis) transferrin: isolation and partial characterization.

The serum transferrin from the primate, Macaca fascicularis is isolated by a purification protocol consisting of ammonium sulphate precipitation and column chromatography. The hexose (galactose + mannose) content of Macaca transferrin is 4.7 mole per mole of protein. Quantitative determination of the sialic acid content shows that there are two sialic acid residues per molecule of Macaca transferrin. This conclusion is supported by the neuraminidase treatment of Macaca transferrin, in which there is a 2-step decrease in electrophoretic mobility. Monoferric Macaca transferrins with Fe3+ selectively labelled at the C- and N-terminal sites (TfFec and FeNTf) are prepared at pH 5.5 and 8.5 using ferric dinitrilotriacetate [Fe(NTA)2] chelate and ferrous ammonium sulphate, respectively.

Animals↗
PubMed1985

Studies on equine transferrin--I. The isolation and partial characterization of the D and R variants.

Each of two genetic variants of equine transferrin, D and R, is isolated from the blood of the heterozygote by a gentle fractionation procedure at pH 7.2. It is shown by step gradient polyacrylamide gel electrophoresis at pH 7.9 that each of these phenotypes exhibits two major bands (designated F, fast, and S, slow) and several minor bands. Components corresponding to these bands are separated by ion-exchange chromatography at pH 6.6 and 6.9 respectively for the D and R variants. The F and S components of each variant contain respectively four and two sialic acid residues. The nature of their heterogeneity is, at least in part, due to their varying sialic acid contents. It has not been possible to desialylate them completely by neuraminidase. On the basis of comparative studies of the tryptic and chymotryptic peptide maps of transferrins D and R it is concluded that there are at least two amino acid substitutions--D:R:Asp:Gly and Glu:Gly. These two substitutions are qualitatively in accordance with the difference in the electrophoretic mobility between the two variants at alkaline pH.

Amino Acid Sequence↗
PubMed1982

The physico-chemical and functional properties of Abarenicola affinis affinis (Ashworth) extracellular haemoglobin (erythrocruorin).

The purified erythrocruorin from the Arenicolid polychaete, Abarenicola affinis affinis (Ashworth) has an S020,w of 57.5 S, and a molecular weight of 3.29 . 10(6) as determined by sedimentation equilibrium at pH 7.0. In the electron microscope it displays a two-tiered hexagonal array characteristic of other annelid erythrocruorins and chlorocruorins, with dimensions of approximately 25.5 nm diameter and 17.0 nm height. The isoelectric point of the molecule is at pH 4.63, and its amino acid composition and absorption spectrum are similar to those of other annelid erythrocruorins. The haem and iron contents are 2.77% and 0.23%, corresponding to minimum molecular weights of 22 250 and 24 800, respectively. In pH-induced dissociation studies, the erythrocruorin is found to be stabilized by Ca2+ up to pH 8.6, while in the presence of EDTA, dissociation begins at pH 7.8. In discontinuous gel electrophoresis, dissociated erythrocruorin gives subunits with molecular weights 263 000, 117 500, 68 400, 35 900 and 28 800, while, in detergent electrophoresis in the presence of 2-mercaptoethanol, polypeptide chains with molecular weights 13 700, 15 800, 24 700, 27 800 and 33 600 are obtained. Abarenicola erythrocruorin is also shown to bind oxygen cooperatively with a Hill coefficient h = 4.0 and P50 = 6.0 mmHg at pH 7.0 and 25 degrees C. Over the pH range of 7.0 to 7.8 the molecule lacks an alkaline Bohr effect.

Amino Acids↗
PubMed1982

The subunits of Abarenicola affinis affinis (Ashworth) extracellular haemoglobin (erythrocruorin).

The subunits of the erythrocruorin of Abarenicola affinis affinis (Ashworth) were investigated by various physicochemical methods. Gel chromatography at pH 9.0 in Sephadex G-200 gave three main protein peaks with molecular weights of 240 000, 100 000 and 31 000, respectively. Polyacrylamide gel electrophoresis showed considerable heterogeneity in the subunits, although the main components gave similar molecular weight ranges to the above. Anomalous results were also obtained in SDS-polyacrylamide gel electrophoresis. In the analytical ultracentrifuge, the subunit from the first peak showed a sedimentation coefficient of 10 S and Mw 250 000, and the second yielded s20,w = 5 S. Functionally, the 10 S subunit showed cooperative oxygen binding, but the Hill coefficient was lower than that observed for the native molecule. A possible model for the subunit structure of Abarenicola affinis affinis erythrocruorin is discussed.

Animals↗
PubMedDate not supplied

Purification and partial characterization of hyaluronidase from stonefish (Synanceja horrida) venom.

1. A marine hyaluronidase was purified 261-fold from the stonefish (Synanceja horrida) crude venom using Sephacryl S-200 HR and heparin affinity-gel chromatography. 2. Stonefish hyaluronidase has a pI of 9.2, a mol. wt of 62,000 and it was purified to a very high spec. act. of 1.6 x 10(6) NFU/mg protein. 3. It was heat sensitive and was inhibited by Cu2+, Hg2+ and heparin. 4. Stonefish hyaluronidase did not contain any haemorrhagic or lethal activity. 5. The N-terminal sequence of stonefish hyaluronidase has been determined to be A-P-S-X-D-E-G-N-K-K-A-D-N-L-L-V-K-K-I-N.

Amino Acid Sequence↗
PubMedDate not supplied

The burnout of nursing staff working with challenging behaviour clients in hospital-based bungalows and a community unit.

The aim of this pilot study was to compare the burnout of nursing staff who worked with clients who displayed challenging behaviour in hospital-based bungalows and a community unit. Twenty-six and 12 nursing staff were randomly chosen from four hospital-based bungalows and a community unit. Staff assessed their clients' behaviour, using the Aberrant Behavior Checklist and were later interviewed, using a self-constructed Staff Questionnaire and the Maslach Burnout Inventory. The results showed that clients in the hospital-based bungalows were more severe in challenging behaviour than those in the community unit. The staff in the hospital-based bungalows were less satisfied with their salaries and enjoyed less their contact with clients. They complained more than the community unit staff and felt that their present training was inadequate. They were also more emotionally exhausted and experienced more depersonalization towards clients than the community unit staff. The level of personal accomplishment was, however, similar in the two groups. Correlations showed that there were significant associations between staff burnout and management issues rather than clients' behaviour, particularly in the hospital-based bungalow group.

Adult↗
PubMedDate not supplied

Proteomic investigation of metabolic shift in mammalian cell culture.

Mammalian cells, under typical cultivation conditions, produce large quantities of lactate and ammonia that affect cell growth adversely and result in low cell concentration. Controlled nutrient feeding to maintain low concentrations of glucose and glutamine reduces metabolite production drastically, altering the metabolism of the cells. This metabolic shift results in higher cell concentration in continuous cultures and does not affect the specific productivity of the cells. We have taken a proteomics approach to investigate the differential protein expression with metabolic shift. Using two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS), we have found at least eight differentially expressed spots; two proteins were down-regulated, and the others were up-regulated with metabolic shift. These included metabolic enzymes, the brain form of phosphoglycerate mutase, which was down-regulated, and the precursor of the 23 kDa subunit of NADH-ubiquinone oxidoreductase, which was up-regulated. Another enzyme, the L1 isozyme of ubiquitin carboxyl-terminal hydrolase, which is involved in protein turnover and degradation, was also up-regulated in the metabolically altered cells. The remaining down-regulated spot had been identified as two isoforms of cytoplasmic actins, while three of the up-regulated spots were viral GAG polyproteins from various murine viruses. An unidentified protein was also up-regulated in the cells with altered metabolic state. This study shows the potential of using a proteomics approach in deciphering the intracellular changes in cells with physiological changes such as metabolism shift. The new insight into cell metabolism afforded by this analysis will greatly facilitate process optimization of continuous cell cultures.

Alkylation↗
PubMedDate not supplied

Reviewing Frankl's Will to meaning and its implications for psychotherapy dealing with post-traumatic stress disorder.

Recent research has shown that people who have gone through war experiences in one way or the other tend to manifest reactions classified as post-traumatic stress disorder. Viktor Frankl, the founder of logotherapy, manifested PTSD reactions when he was in a concentration camp. This paper attempts to sketch how he lived through his traumatic experiences by relying on the principle of Will to Meaning as his survival mechanism. Two implications will be outlined derived from this principle for psychotherapies dealing with disaster survivors.

Behavior Therapy↗