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Biomedical subjects

K S Cheah

Publications and source records attributed to K S Cheah.

At least 55 records · Page 3Linked to original sources
PubMed1988

Lipid analysis of skeletal muscle from pigs susceptible to malignant hyperthermia.

Previous studies demonstrated that lipid profiles of humans and pigs susceptible to malignant hyperthermia (MH) differ from those of normal humans and pigs. Lipid extraction techniques retaining in vivo lipid profiles most closely were used in the present study to determine if stimulation of lipolysis by the processes of homogenization or extraction might account for the reported differences in lipid profiles. No differences were observed among three genotypes of British Landrace pigs with respect to cholesterol levels, triglyceride levels, or total lipid phosphorus values of whole muscle (longissimus dorsi). Phospholipid distributions were the same for all three groups. Individual free fatty acids and fatty acids acylated to triglycerides were similar among the genotypes. These results do not support altered lipid profiles in vivo in MH-susceptible swine. Previously used homogenization and extraction procedures most likely affect the lipolytic activity to a different extent in muscle from MH-susceptible pigs and normal pigs.

Animals↗
PubMed1987

Tissue-specific expression of the human type II collagen gene in mice.

Type II collagen is crucial to the development of form in vertebrates as it is the major protein of cartilage. To study the factors regulating its expression we introduced a cosmid containing the human type II collagen gene, including 4.5 kilobases of 5' and 2.2 kilobases of 3' flanking DNA, into embryonic stem cells in vitro. The transformed cells contribute to all tissues in chimeric mice allowing the expression of the exogenous gene to be studied in vivo. Human type II collagen mRNA is restricted to tissues showing transcription from the endogenous gene and human type II collagen is found in extracellular matrix surrounding chondrocytes in cartilage. The results indicate that the cis-acting requirements for correct temporal and spatial regulation of the gene are contained within the introduced DNA.

Animals↗
PubMed1986

The human type II collagen gene (COL2A1) assigned to 12q14.3.

A cosmid clone containing the entire human type II alpha 1 collagen gene (COL2A1) was used as probe in the Southern analysis of DNA from a panel of human/hamster somatic cell hybrids containing different portions of human chromosome 12. Two of the hybrids exhibited a similar terminal deletion q14.3----qter, but one was positive for the gene while the other was negative. Therefore, the gene must reside in the region q14.3.

Animals↗
PubMed1985

A highly polymorphic region 3' to the human type II collagen gene.

We have characterised a highly polymorphic region 1.3kb downstream of the human Type II collagen gene. It consists of a highly AT-rich tandem repetitive region (minisatellite) approximately 650bp long. Two alleles had been observed previously, differing in size by approximately 300bp. When this region was cloned from four unrelated individuals carrying the larger allele, DNA sequence data identified three alleles, suggesting far higher polymorphism than was originally supposed. This minisatellite was shown to be present in a single copy in the human genome, and to have arisen after the divergence of Old and New World monkeys.

Alleles↗
PubMed1985

Malignant hyperthermia: molecular defects in membrane permeability.

Malignant hyperthermia (MH), a genetically inherited disorder of skeletal muscle, is due to molecular defect in membrane permeability. The alteration in membrane permeability is suggested to be due to enhanced phospholipase A2 activity which is responsible for the increased level in sarcoplasmic Ca2+. The excess Ca2+ is responsible for muscle hyper-rigidity and enhanced rate of glycolysis, resulting in a rapid rate of lactic acid production and a low pH in MH muscle.

Animals↗
PubMed1985

Influence of environmental temperature and energy intake on porcine skeletal muscle mitochondria.

The influence of acclimation to a cold (10 degrees C) or warm (35 degrees C) environment on the functional characteristics of skeletal muscle mitochondria has been investigated in young pigs on a high (H) or low (L) energy intake. Living at 10 degrees C increased the amount of mitochondrial protein and the concentration of cytochrome. Ca2+-stimulated succinate oxidation by mitochondria from the 35H group was tightly coupled and similar to that in pigs living under normal husbandry conditions. Mitochondria from the three other groups were readily uncoupled and thus resembled those from pigs susceptible to malignant hyperthermia. Arrhenius plots of State 3 Ca2+-stimulated succinate oxidation showed that energy intake altered the transition temperature suggesting possible differences in the structure of the mitochondrial membrane.

Adenosine Diphosphate↗
PubMed1985

Identification and characterization of the human type II collagen gene (COL2A1).

The gene contained in the human cosmid clone CosHcol1, previously designated an alpha 1(I) collagen-like gene, has now been identified. CosHcol1 hybridizes strongly to a single 5.9-kilobase mRNA species present only in tissue in which type II collagen is expressed. DNA sequence analysis shows that this clone is highly homologous to the chicken alpha 1(II) collagen gene. These data together suggest that CosHcol1 contains the human alpha 1(II) collagen gene COL2A1. The clone appears to contain the whole gene (30 kilobases in length) and will be extremely useful in the study of cartilage development and for identifying those inherited chondrodystrophies in which defects occur in this gene.

Animals↗
PubMed1984

Lethal osteogenesis imperfecta congenita and a 300 base pair gene deletion for an alpha 1(I)-like collagen.

Broad boned lethal osteogenesis imperfecta is a severely crippling disease of unknown cause. By means of recombinant DNA technology a 300 base pair deletion in an alpha 1(I)-like collagen gene was detected in six patients and four complete parent-child groups including patients with this disease. One from each set of the patients' clinically unaffected parents also carried the deletion, implying that affected patients were genetic compounds. The study suggests that prenatal diagnosis should be possible with 100% accuracy in subjects without the deletion and with 50% accuracy in those who possess it (who would be either heterozygous--normal, or affected with the disease).

Base Composition↗
PubMed1983

Effect of trifluoperazine on skeletal muscle mitochondrial respiration.

The effect of trifluoperazine on the respiration of porcine liver and skeletal muscle mitochondria was investigated by polarographic and spectroscopic techniques. Low concentrations of trifluoperazine (88 nmol/mg protein) inhibited both the ADP- and Ca2+-stimulated oxidation of succinate, and reduced the values of the respiratory control index and the ADP/O and Ca2+/O ratio. High concentrations inhibited both succinate and ascorbate plus tetramethyl-p-phenylenediame (TMPD) oxidations, and uncoupler (carbonyl cyanide p-trifluromethoxyphenylhydrazone) and Ca2+-stimulated respiration. Porcine liver mitochondria were more sensitive to trifluoperazine than skeletal muscle mitochondria. Trifluoperazine inhibited the electron transport of succinate oxidation of skeletal muscle mitochondria within the cytochrome b-c1 and cytochrome c1-aa3 segments of the respiratory chain system. 233 nmol trifluoperazine/mg protein inhibited the aerobic steady-state reduction of cytochrome c1 by 92% with succinate as substrate, and of cytochrome c and cytochrome aa3 by 50-60% with ascorbate plus TMPD as electron donors. Trifluoperazine can thus inhibit calmodulin-independent reactions particularly when used at high concentrations.

Adenosine Diphosphate↗
PubMed1982

Isolation and characterization of a human collagen alpha 1(I)-like gene from a cosmid library.

We have isolated a human collagen alpha 1(I)-like gene from a cosmid library. The clone which contains 37kb of human DNA has been shown to contain this gene by DNA sequencing, hybrid arrest and hybrid selection assays and Northern blot hybridizations. The collagen gene sequence extends through most of the cloned DNA and must, therefore, be at least 35kb in length.

Base Sequence↗
PubMed1981

Mitochondrial calcium transport and calcium-activated phospholipase in porcine malignant hyperthermia.

The interaction of Ca2+ with mitochondria isolated from longissismus dorsi, a predominantly white skeletal muscle, of normal and malignant hyperthermia pigs was investigated using tightly-coupled preparations. Arrhenius plots of mitochondrial Ca2+ -stimulated respiration for succinate oxidation of malignant hyperthermia pigs showed a transition temperature (Tt) of 26.31 +/- 0.80 degrees C (n = 5), which was decreased by spermine to 15.41 +/- 0.69 degrees C (n = 3), a value very similar to that for normal pigs. No difference in either the Tt or in the activation energy (Ea) was observed between the two types of pigs when ADP was used instead of Ca2+. Mitochondria of malignant hyperthermia pigs were uncoupled at 40 degrees C by exogenous Ca2+ at 1221 +/- 301 (n = 9) nmol Ca2+ per mg proteinn during succinate oxidation and the uncoupled mitochondria showed large amplitude swelling. Both the Ca2+ -induced uncoupling and swelling were prevented by bovine serum albumin and by the phospholipase inhibitors, spermine and tetracaine. In contrast, mitochondria of normal pigs were still tightly coupled even after a total addition of 2313 +/- 287 (n = 5) nmol Ca2+ per mg protein and retained the original condensed configuration in the presence or absence of spermine and tetracaine. Mitochondria of malignant hyperthermia pigs contained significantly (P less than 0.001) higher quantities of endogenous Ca2+ and showed a significantly (P less than 0.001) faster FCCP-induced endogenous Ca2+ efflux rate than normal when monitored spectroscopically with murexide. No significant difference was observed in either the rate of exogenous Ca2+ uptake or in the extent of Ca2+ accumulated in the aerobic steady state during succinate oxidation between the two types of pigs. The rate of mitochondrial Ca2+ efflux of malignant hyperthermia pigs during anaerobiosis was about twice that of normal. Experimental evidence suggests that mitochondria from musculi longissimus dorsi of malignant hyperthermia pigs contained a Ca2+ -stimulated phospholipase A2 (EC 3.1.1.4, phosphatide 2-acylhydrolase), and that this enzyme if present in mitochondria of normal pigs is either latent or in very low concentration. The significance of the Ca2+ -stimulated phospholipase A2 and its association with the enhanced rate of glycolysis in porcine malignant hyperthermia syndrome and in the post-mortem formation of the pale, soft and exudative conditions observed in white skeletal muscles of malignant hyperthermia pigs is discussed.

Adenosine Diphosphate↗
PubMed1981

Identification of the primary translation product of elastic mRNA.

Poly (A) RNA was prepared from matrix free cells derived from the major thoracic arteries of 17-day old chick embryos. This was translated in a messenger-dependent reticulocyte cell-free system in the presence of [35S]-methionine, [3H]-proline or [3H]-valine. The translation products were analyzed by SDS polyacrylamide gel electrophoresis and fluorography and elastin-related products were immunoprecipitated by the addition of anti-elastin sheep antiserum in the presence of proteinase inhibitors and the precipitate analyzed by the same technique. The elastin-related products were found to consist of a doublet of two closely related polypeptides (mol. wts. 70,000 and 73,000) comigrating with a tropoelastin standard. Unhydroxylated pro alpha 1 and pro alpha 2 collagen polypeptides were also found, but no elastin related product corresponding to proelastin. It is concluded that the primary translation product of elastin mRNA consists of two polypeptides between 70,000 and 73,000 mol wt. which are closely related in size and immunoreactivity to tropoelastin.

Animals↗