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Biomedical subjects

M C Chung

Publications and source records attributed to M C Chung.

At least 37 records · Page 2Linked to original sources
PubMed1999

Amino acid sequence of trocarin, a prothrombin activator from Tropidechis carinatus venom: its structural similarity to coagulation factor Xa.

Among snake venom procoagulant proteins, group II prothrombin activators are functionally similar to blood coagulation factor Xa. We have purified and partially characterized the enzymatic properties of trocarin, the group II prothrombin activator from the venom of the Australian elapid, Tropidechis carinatus (rough-scaled snake). Prothrombin activation by trocarin is enhanced by Ca2+, phospholipids, and factor Va, similar to that by factor Xa. However, its amidolytic activity on peptide substrate S-2222 is significantly lower. We have determined the complete amino acid sequence of trocarin. It is a 46,515-Dalton glycoprotein highly homologous to factor Xa and shares the same domain architecture. The light chain possesses an N-terminal Gla domain containing 11 gamma-carboxyglutamic acid residues, followed by two epidermal growth factor (EGF)-like domains; the heavy chain is a serine proteinase. Both chains are likely glycosylated: the light chain at Ser 52 and the heavy chain at Asn 45. Unlike other types of venom procoagulants, trocarin is the first true structural homologue of a coagulation factor. It clots snake plasma and thus may be similar, if not identical, to snake blood coagulation factor Xa. Unlike blood factor Xa, it is expressed in high quantities and in a nonhepatic tissue, making snake venom the richest source of factor Xa-like proteins. It induces cyanosis and death in mice at 1 mg/kg body weight. Thus, trocarin acts as a toxin in venom and a similar, if not identical, protein plays a critical role in hemostasis.

Amino Acid Sequence↗
PubMed1999

Rhodocetin, a novel platelet aggregation inhibitor from the venom of Calloselasma rhodostoma (Malayan pit viper): synergistic and noncovalent interaction between its subunits.

A novel platelet aggregation inhibitor, rhodocetin, was purified from the crude venom of Calloselasma rhodostoma. It inhibited collagen-induced platelet aggregation in a dose-dependent manner, with an IC50 of 41 nM. Rhodocetin has a heterodimeric structure with alpha and beta subunits, which could be separated on a nonreducing denaturing gel or reverse-phase HPLC column. Individually neither subunit inhibited platelet aggregation even at 2.0 microM concentration. Titration and reconstitution experiments showed that, when these subunits are mixed to give a 1:1 complex, most of its biological activity was recovered. The reconstituted complex inhibited platelet aggregation with an IC50 of 112 nM, about 3-fold less effective than the native molecule. Circular dichroism analysis revealed that the reconstituted complex had a spectrum similar to that of the native protein. By using surface plasmon resonance studies, we established that the stoichiometry of binding between the two subunits is 1:1 and the subunits interact with a Kd of 0.14 +/- 0.04 microM. The complete amino acid sequences of the alpha (15956.16 Da, 133 residues) and beta (15185.10 Da, 129 residues) subunits show a high degree of homology with each other (49%) and with the Ca2+-dependent lectin-related proteins (CLPs) (typically 29-48%) isolated from other snake venoms. Unlike the other members of the family in which the subunits are held together by an interchain disulfide bond, rhodocetin subunits are held together only through noncovalent interactions. The cysteinyl residues forming the intersubunit disulfide bridge in all other known CLPs are replaced by Ser-79 and Arg-75 in the alpha and beta subunits of rhodocetin, respectively. These studies support the noncovalent and synergistic interactions between the two subunits of rhodocetin. This is the first reported CLP dimer with such a novel heterodimeric structure.

Amino Acid Sequence↗
PubMed1999

Identification of Enterobacteriaceae bacteria by direct matrix-assisted laser desorptiom/ionization mass spectrometric analysis of whole cells.

Several members of Enterobacteriaceas were analyzed by matrix-assisted laser desorption ionization (MALDI) time-of-flight mass spectrometry (TOFMS). Characteristic mass spectral peaks and patterns were observed in the mass range of 2 to 20 kDa. The mass peaks reported to be reproducibly observed by previous researchers, which were claimed to serve as species/strain-specific biomarkers, are consistently observed in our current study. Despite the high degree of similarity found in the MALDI mass spectra within the enteric bacteria, minor yet notable differences existed to allow their differentiation. Five spectral peaks at m/z 4364, 5380, 6384, 6856, and 9540, generated reproducibly for each genus studied here, are assigned as family-specific biomarkers for the Family Enterobacteriaceae. The mass peaks at m/z 7324, 7724, 9136, and 9253 are assigned as genus-specific biomarkers for Salmonella. Some unique biomarkers characterizing the species and strains of E. coli are also presented.

Biomarkers↗
PubMed1999

The relationship between trauma and personality in victims of the Boeing 737-2D6C crash in Coventry.

The aims of this paper were to (a) ascertain the extent of psychological distress and (b) identify the association between personality variables and psychological distress among individuals who had been exposed to an aircraft disaster in Coventry, U.K. Hundreds of people escaped death but were exposed to the impact of the disaster when a Boeing 737-2D6C 7T-VEE crashed into a woodland area on the edge of a large housing estate in Coventry, U.K. in 1994. Eighty-two residents were randomly chosen for interviews in which they were assessed using the Impact of Event Scale, the General Health Questionnaire, and the Eysenck Personality Questionnaire-R Short Scale (EPQ-R). The results showed that the Coventry residents' scores reached similar levels of intrusion and avoidance compared with standardized samples and the Lockerbie samples. Fifty-two percent reached the GHQ case level score, which was again similar to the Lockerbie residents. The Coventry residents were significantly less extroverted and neurotic than standardized samples. Stepwise multiple regression showed that there were associations between intrusion and neuroticism and intrusion and extroversion, as well as between avoidance and neuroticism.

Adult↗
PubMed1999

Nursing students' perceptions of their first mental health placement.

The way in which the theory and practice of mental health nursing is presented to students is of particular importance, especially at a time when recruitment into the profession is falling. This study sought to examine how nursing students experience their first mental health placement. A pre-/post-test design was used with a standardized instrument (Attitude Towards Psychiatry questionnaire) to assess the influence of their mental health placements on nursing students' attitudes towards mental health nurses, mental health nursing practice and education and theoretical aspects of mental health nursing. Results suggest that the placements were effective in improving students' regard in each of these areas. Though no generalizable conclusions can be inferred from this study some interesting insights are provided by nursing students into their perceptions of current mental health nursing practice.

Adult↗
PubMed1999

Describing traumatic responses and distress of community residents directly and indirectly exposed to an aircraft crash.

This study described the traumatic responses and the extent of psychological distress among residents who had been exposed directly or indirectly to an aircraft crash in Coventry, U.K. The direct exposure group consisted of 62 residents who were on the housing estate at the time of the accident and 20 who were not. They were interviewed on their subjective responses to the crash and then asked to fill in two distress measures: the Impact of Event Scale (IES) and the General Health Questionnaire (GHQ). The results showed a contrast between the responses of the two groups at the time of the crash. They also showed that the direct exposure group had a significantly higher score in the IES item of avoidance, the IES total, the GHQ items of somatization, social dysfunction, and the total score than the indirect exposure group had. Associations were then made between subjective responses and distress measures. Stepwise multiple regression analyses showed that for the direct exposure residents, the IES total was predicted by "whether they received professional help" and "whether they were worried about their safety after the crash." The GHQ total of the direct exposure residents was predicted by "anger about what had happened to them." For the indirect exposure residents, the IES total was predicted by "their present feelings when they heard planes flying over." There were no predictions found between responses and the GHQ total.

Accidents, Aviation↗
PubMed1999

A follow-up study of mentally disordered offenders after a court diversion scheme: six-month and one-year comparison.

The aim of this study was to investigate offenders' living patterns, quality of life, types of aftercare received and psychological well-being, following their diversion from one court diversion scheme in England. Sixty-five offenders were followed up after six months, 22 of whom were also followed up after one year. Their transient living patterns throughout these two periods were described. Their quality of life was found to be poor in that most did not have an occupation, nor did they receive education, and the life experiences during both periods were lower than that of standardized urban samples. There were no significant changes in their poor life experiences over the two periods. Over 55% of the 22 offenders still contacted their GPs regularly but there was a significant drop in those consulting hospital doctors. Only a small number of offenders had contact with the social services in these two periods and no significant differences were found between them. Their GHQ total scores went above cutoff point and the level of psychiatric conditions was similar in both periods.

Adolescent↗
PubMed1998

A description of a forensic diversion service in one city in the United Kingdom.

This paper describes some recent research on forensic diversion services and the application of such services in Birmingham, UK. Key components include the diversion scheme in court, diversion at point of arrest, and the remand prison liaison scheme. Various community facilities, such as a specialized bail hostel and landlady schemes to which offenders are possibly referred, are also briefly described. We advocate the need for further research in order to examine the effectiveness of the services in Birmingham.

England↗
PubMed1998

Longitudinal changes in adaptive behavior in adults with Down syndrome: interim findings from a longitudinal study.

Age-related changes in adaptive behavior have been established in adults with Down syndrome. However, most studies have been cross-sectional, without controls for cohort and survival effects. We examined underlying factors for age-related decline in adaptive behavior in 128 adults with trisomy 21 over a 3-year period. Presence of dementia was the only determining factor, although the difference in trend over time as compared to subjects without dementia was not significant. No association between gender, sensory loss, severity of mental retardation, or place of residence was found. For subjects with no significant medical disorder (physical or psychological), no decline was seen. This longitudinal study confirms previously reported findings.

Activities of Daily Living↗
PubMed1997

Genomic structure of a potassium channel toxin from Heteractis magnifica.

We provide information on the gene encoding the K+ channel toxin, HmK, of the sea anemone Heteractis magnifica. A series of DNA amplifications by PCR, which included the amplification of the 5'-untranslated region of the gene, showed that an intron of 402 nucleotides separated the sequence that encodes the matured toxin from the signal peptide sequence. A second 264 nucleotide intron interrupted the 5'-untranslated region of the previously reported HmK cDNA. Two possible transcription-initiation sites were identified by primer extension analysis. Corresponding TATA-box consensus sequences, characteristic of a promoter region, were also located from PCR products of uncloned libraries of adaptor-ligated genomic DNA fragments. The coding region for matured HmK is intronless. The same is also true for other sea anemone toxins reported thus far. More notably, a similar intron-exon organization is present in other ion channel-blocking toxins from scorpions implying that molecules having similar functions share a similar organization at the genomic level suggesting a common path of evolution.

Amino Acid Sequence↗
PubMed1997

A new potassium channel toxin from the sea anemone Heteractis magnifica: isolation, cDNA cloning, and functional expression.

A new potassium channel toxin, HmK, has been isolated from the sea anemone Heteractis magnifica. It inhibits the binding of [125I]-alpha-dendrotoxin (a ligand for voltage-gated K channels) to rat brain synaptosomal membranes with a Ki of about 1 nM, blocks K+ currents through Kv 1.2 channels expressed in a mammalian cell line, and facilitates acetylcholine release at the avian neuromuscular junction. HmK comprises of 35 amino acids (Mr 4055) with the sequence R1TCKDLIPVS10ECTDIRCRTS20MKYRLNLCRK30TCGSC35. A full assignment of the disulfide linkages was made by using partial reduction with tri(2-carboxyethyl)phosphine (TCEP) at acid pH and rapid alkylation with iodoacetamide. The disulfide bridges were identified as Cys3-Cys35, Cys12-Cys28, and Cys17-Cys32. A cDNA clone encoding HmK was isolated using RT-PCR from the total RNA obtained from sea anemone tentacles, while the 5'- and 3'-flanking regions of the cDNA were amplified by RACE. The full-length cDNA was 563 bp long and contained a sequence encoding a signal peptide of 39 amino acids. The coding region for matured HmK toxin was cloned and expressed as a glutathione S-transferase (GST) fusion product in the cytoplasm of Escherichia coli. After affinity purification and cleavage, the recombinant toxin was shown to be identical to native HmK in its N-terminal sequence, chromatographic behavior, and binding to dendrotoxin binding sites on rat brain membranes.

Amino Acid Sequence↗
PubMed1997

Rabbit ubiquitin-activating enzyme E1: cDNA cloning, sequence and expression.

A cDNA clone encoding ubiquitin-activating enzyme E1 has been isolated from a rabbit heart cDNA library and sequenced. The 3.485 kb cDNA contains an open reading frame of 1058 amino acid residues which predicts a protein of approx. 118 kDa. The deduced protein sequence exhibits a very high homology to other ubiquitin-activating enzymes identified in a variety of organisms. Northern blot analysis reveals a single transcript of approx. 3.5 kb in all the rabbit tissues examined. The entire coding region of the rabbit E1 cDNA has been expressed as a his-tagged protein. The recombinant protein has been verified by its ability to cross-react with anti-human E1 antibodies. Ubiquitin thiolester assay shows that the recombinant rabbit E1 protein is functional.

Amino Acid Sequence↗
PubMed1997

Makatoxin I, a novel toxin isolated from the venom of the scorpion Buthus martensi Karsch, exhibits nitrergic actions.

Buthus martensi Karsch venom exhibits nitrergic action in rat anococcygeus muscle (ACM). We have purified a novel toxin, makatoxin I (MkTx I), which exhibits nitrergic action, to homogeneity from this venom by a combination of gel-filtration, cation-exchange chromatography, and reverse-phase chromatography. Its purity was assessed by capillary electrophoresis and mass spectrometry. Its molecular weight was found to be 7031.71 +/- 2.88 as calculated from electrospray mass spectrographic data. The complete amino acid sequence was elucidated by sequencing of reduced and S-pyridylethylated toxin and a carboxyl-terminal peptide, P55-64, generated by the cleavage of toxin with endoproteinase Lys-C. The complete sequence of MkTx I is GRDAYIADSENCTYTCALNPYCNDLCTKNGAKSGYCQWAGRYGNACWCIDLPDKVPIRISGSCR. This toxin is composed of 64 amino acid residues and contains 8 half-cystine residues. Structurally, MkTx I has high similarity to Bot I and Bot II when compared with toxins from other scorpion species. The effects of MkTx I on nitrergic responses were investigated using the rat isolated ACM mounted in Krebs solution (37 degrees C, 5% CO2 in O2). MkTx I (2 microg/ml) markedly relaxed the carbachol-precontracted ACM; the relaxation was inhibited by the stereoselective inhibitor of nitric oxide synthase, Nomega-nitro-L-arginine methyl ester (50 microM). Thus, MkTx I is the first alpha-toxin that can mediate nitrergic responses in the rat isolated ACM.

Amino Acid Sequence↗
PubMed1997

Cloning, characterization and expression of a cDNA clone encoding rabbit ubiquitin-conjugating enzyme, E2(32k).

A cDNA clone encoding rabbit E2(32k) was obtained by library screening and PCR. The cDNA contains an open reading frame coding for 238 amino acids which shows an overall identity of 81% to human CDC34, the cell cycle-related ubiquitin-conjugating enzyme. A 50% homology to yeast CDC34 within the conserved core domain was also observed. Northern blot analysis indicated that three transcripts existed in all six rabbit tissues examined but their expression levels varied over a wide range. The putative cDNA coding region was highly expressed in Escherichia coli as a his-tagged protein which was purified to homogeneity. The ability of this expressed protein to form a thiolester bond with ubiquitin showed that it was functionally active. The ability of this protein to catalyze the conjugation of ubiquitin to histone H2A and H2B was also examined.

Amino Acid Sequence↗
PubMed1997

Complete amino acid sequence of an acidic, cardiotoxic phospholipase A2 from the venom of Ophiophagus hannah (King Cobra): a novel cobra venom enzyme with "pancreatic loop".

A phospholipase A2 (OHV A-PLA2) from the venom of Ophiophagus hannah (King cobra) is an acidic protein exhibiting cardiotoxicity, myotoxicity, and antiplatelet activity. The complete amino acid sequence of OHV A-PLA2 has been determined using a combination of Edman degradation and mass spectrometric techniques. OHV A-PLA2 is composed of a single chain of 124 amino acid residues with 14 cysteines and a calculated molecular weight of 13719 Da. It contains the loop of residues (62-66) found in pancreatic PLA2s and hence belongs to class IB enzymes. This pancreatic loop is between two proline residues (Pro 59 and Pro 68) and contains several hydrophilic amino acids (Ser and Asp). This region has high degree of conformational flexibility and is on the surface of the molecule, and hence it may be a potential protein-protein interaction site. A relatively low sequence homology is found between OHV A-PLA2 and other known cardiotoxic PLA2s, and hence a contiguous segment could not be identified as a site responsible for the cardiotoxic activity.

Amino Acid Sequence↗
PubMed1997

Role of polyethyleneimine in the purification of recombinant human tumour necrosis factor beta.

The chromatographic behaviour of recombinant human tumour necrosis factor beta (rhTNF-beta) (pI approximately 9.0) during cation-exchange chromatography at pH 7.5 is investigated. Without prior treatment of the Escherichia coli cell extract with polyethyleneimine (PEI), very little rhTNF-beta was bound to the column. However, upon addition of 5% PEI (100 microliters ml-1) to the cell lysate, rhTNF-beta was shown to bind to cation-exchange columns normally. TNF-beta was readily precipitated from the clarified cell extract by 20% ammonium sulphate, but ony ca. 25% of this precipitate could be re-solubilized for further purification. However, when 5% PEI was included in the solubilization buffer, the balance of the rhTNF-beta could be recovered. It is proposed that charge interaction between rhTNF-beta and nucleic acids in the cell extract is responsible for both of these anomalous phenomena, and that PEI (a cationic polyelectrolyte) was able to disrupt this interaction by displacing rhTNF-beta from the charge complex.

Ammonium Sulfate↗
PubMed1997

Cloning and characterization of cDNAs encoding three isoforms of phospholipase A2 in Malayan spitting cobra (Naja naja sputatrix) venom.

cDNAs encoding three phospholipase A2 (PLA2) isoforms in Naja naja sputatrix were cloned and characterized. One of them encoded an acidic PLA2 (APLA) while the others encoded neutral PLA2 (NPLA-1 and NPLA-2). The specific characteristics of APLA and NPLA were attributed to mutations at nt139 and nt328 from G to C and G to A, respectively, resulting in amino acid substitutions from Asp20 and 83 in APLA to His20 and Asn83 in NPLA. Amino acid sequencing of purified protein also showed the presence of this Asp20 and His20 in APLA and NPLA, respectively. The cDNA encoding one of the PLA2 (NAJPLA-2A), when expressed in Escherichia coli, yielded a protein that exhibited PLA2 activity.

Amino Acid Sequence↗