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Biomedical subjects

C Holm

Publications and source records attributed to C Holm.

At least 145 records · Page 8Linked to original sources
PubMed1988

Hormone-sensitive lipase: sequence, expression, and chromosomal localization to 19 cent-q13.3.

Hormone-sensitive lipase, a key enzyme in fatty acid mobilization, overall energy homeostasis, and possibly steroidogenesis, is acutely controlled through reversible phosphorylation by catecholamines and insulin. The 757-amino acid sequence predicted from a cloned rat adipocyte complementary DNA showed no homology with any other known lipase or protein. The activity-controlling phosphorylation site was localized to Ser563 in a markedly hydrophilic domain, and a lipid-binding consensus site was tentatively identified. One or several messenger RNA species (3.3, 3.5, or 3.9 kilobases) were expressed in adipose and steroidogenic tissues and heart and skeletal muscle. The human hormone-sensitive lipase gene mapped to chromosome 19 cent-q13.3.

Amino Acid Sequence↗
PubMed1988

Effects of dopamine on the portal circulation after therapeutic hepatic artery ligation.

The effects of exogenous dopamine (2, 4 and 6 micrograms.kg-1.min-1 i.v.) on the portal circulation were studied in six patients following therapeutic hepatic artery ligation. Portal blood flow (PBF) was measured by the continuous thermodilution technique. Portal venous pressure (PVP, n = 3) was monitored through the thermodilution catheter to allow derivation of preportal vascular resistance (PVR). Blood samples were taken through the portal venous catheter for measurement of dopamine. A significant increase in PBF and a decrease in PVR were observed during graded i.v. dopamine infusion. Thus, PBF was 961 +/- 119 ml.min-1 during control conditions and increased to 1446 +/- 221 ml.min-1 during the dopamine infusion at 6 micrograms.kg-1.min-1. No significant changes in mean arterial pressure or PVP were observed during dopamine administration. The pharmacokinetics of dopamine did not differ from that previously reported in patients with an intact arterial supply. In conclusion, our data indicate that exogenous dopamine consistently increases PBF by preportal vasodilation, also in patients with a surgically restricted hepatic arterial blood supply.

Adult↗
PubMed1988

Bone scintigraphy in post-traumatic reflex dystrophy.

Twenty-five patients presenting consecutively with persistent pain and tenderness, edema, and vasomotor instability in an extremity following trauma to the same extremity were clinically diagnosed as having early post-traumatic reflex dystrophy (PRD). All underwent scintigraphy with 99mTechnetium methylene diphosphonate before institution of prednisolone treatment. The time lag between the occurrence of the initial trauma and scintigraphic imaging averaged 11.5 weeks. Of 25 scintigrams, 24 showed multiple periarticular accentuation in the affected extremity. Scintigraphy is a useful clinical aid to establish the diagnosis of early PRD.

Adult↗
PubMed1987

Immunological evidence for the presence of hormone-sensitive lipase in rat tissues other than adipose tissue.

A polyclonal rabbit antibody was used to detect hormone-sensitive lipase in rat organs other than white adipose tissue. Inhibition of tissue diacylglycerol lipase activity by the anti-hormone-sensitive lipase, and by NaF, Hg2+ and diisopropyl fluorophosphate, known inhibitors of the hormone-sensitive lipase, demonstrated its presence in the adrenals, ovaries, testes, heart and skeletal muscle, but not in the liver and kidneys. After enrichment by immunoprecipitation an immunoreactive protein, corresponding to the adipose tissue hormone-sensitive lipase 84 kDa subunit, and some additional, higher Mrapp proteins, were detected by Western blotting in the same tissues. The adipose tissue contained greater than 80% of the total hormone-sensitive lipase, with 5-10- and 50-100-fold lower specific activity in the steroid-producing and the muscle tissues, respectively.

Adipose Tissue↗
PubMed1987

Hormone-sensitive lipase in brown adipose tissue: identification and effect of cold exposure.

Hormone-sensitive lipase (HSL) in brown adipose tissue from mice was identified through immunoprecipitation with a polyclonal antibody (anti-HSL) towards rat white fat HSL and Western blotting. An 82 kDa polypeptide, slightly smaller than the rat white fat HSL 84 kDa subunit, was detected and its identity as HSL verified by inhibition properties. The HSL concentration per g tissue was several-fold higher in the mouse brown adipose tissue than in the rat white adipose tissue, but the specific activities per mg protein were similar. Cold-exposure (4 degrees C) of the mice for 24 h approximately doubled the HSL concentration but this increase parallelled the overall protein increase and did not reflect a specific effect on the HSL.

Adipose Tissue, Brown↗
PubMed1986

Demonstration of the amphiphilic character of hormone-sensitive lipase by temperature-induced phase separation in Triton X-114 and charge-shift electrophoresis.

Temperature-induced phase separation in Triton X-114 (Bordier, C. (1981) J. Biol. Chem. 256, 1604-1607) and charge-shift electrophoresis (Helenius, A., and Simons, K. (1977) Proc. Natl. Acad. Sci. U. S. A. 74, 529-532) were used to examine the amphiphilic character of hormone-sensitive lipase, purified from rat adipose tissue. In contrast to ATP-citrate lyase, a reference hydrophilic protein, the lipase was shown to partition predominantly (approximately 80%) into the detergent-rich phase upon phase separation in Triton X-114. Furthermore, its electrophoretic mobility was markedly shifted anodally and cathodally upon charge-shift electrophoresis in the presence of sodium taurodeoxycholate and cetyltrimethylammonium bromide, respectively. The results demonstrate that hormone-sensitive lipase possesses detergent-binding hydrophobic domain(s) and exhibits the same amphiphilicity as typical intrinsic membrane proteins.

ATP Citrate (pro-S)-Lyase↗
PubMed1986

Portal blood flow in man during graded positive end-expiratory pressure ventilation.

The cardiovascular response to graded PEEP ventilation (5-10 cm H20) was studied peroperatively in patients undergoing cholecystectomy (n = 8) or hepatic tumour surgery (n = 3). Portal blood flow was measured by the continuous thermodilution technique and cardiac output, in a sub-group of the patients, by impedance cardiography. A parallel reduction in cardiac output and portal blood flow was demonstrated in patients undergoing cholecystectomy as the result of the application of PEEP. Thus, ventilation with 5 cm H2O of PEEP elicited a 17% decrease in cardiac output and a 26% decrease in portal blood flow. During 10 cm H2O of PEEP cardiac output decreased by 22% and portal blood flow by 32%. However, there were no significant changes in preportal tissue perfusion pressure by the application of PEEP and preportal vascular resistance increased by 22% and 30%, respectively. This indicates that a vasoconstrictor response, elicited by PEEP, in the preportal tissue is the predominating mechanism for the observed decrease in portal blood flow. Systemic oxygen transport decreased by 214 ml/min during PEEP ventilation, but preportal tissue oxygen utilization was not significantly changed due to a concurrent increase (2.9%; p less than 0.05) in oxygen extraction.

Adolescent↗
PubMed1986

A rapid, efficient method for isolating DNA from yeast.

A method is described for the purification of chromosomal and plasmid DNA from the yeast Saccharomyces cerevisiae. This method is rapid, gives 75% of theoretical yield, and produces DNA that can be cut with restriction endonucleases. Yeast cells are treated with zymolyase, and the resulting spheroplasts are lysed in the presence of the chaotropic agent guanidine hydrochloride. After a brief ethanol precipitation, protein is removed by treatment with proteinase K followed by phenol-chloroform extraction. After ethanol precipitation, the DNA is sufficiently pure for restriction analysis or for the transformation of Escherichia coli.

Chromosomes↗
PubMed1985

DNA topoisomerase II is required at the time of mitosis in yeast.

We have constructed five new temperature-sensitive DNA topoisomerase II mutations and have analyzed their physiological consequences in yeast. Several lines of evidence suggest that the activity of topoisomerase II is required specifically at the time of miosis. First, top2 mutations cause dramatic lethality at the restrictive temperature, but only if the mutant cells are actively traversing the cell cycle. Second, temperature-shift experiments with synchronized cultures show that the onset of inviability coincides with the time of mitosis. Third, fluorescence microscopy reveals that the normal progression of mitosis is disturbed in mutant cells at the restrictive temperature. Finally, inviability at the restrictive temperature is prevented by nocodazole, an inhibitor of tubulin polymerization that prevents formation of the mitotic spindle. These results are consistent with the hypothesis that the essential function of topoisomerase II is to allow the separation of intertwined chromosomal DNA molecules during mitosis.

Benzimidazoles↗
PubMed1984

Purification of mouse brain (Na+ + K+)-ATPase catalytic unit, characterization of antiserum, and immunocytochemical localization in cerebellum, choroid plexus, and kidney.

The denatured catalytic polypeptide of mouse brain (Na+ + K+)-adenosine triphosphatase(ATPase) was separated from microsomal membranes on polyacrylamide gels and used as an immunogen. The antiserum, characterized by immunoblots, recognizes the polypeptide corresponding to the catalytic unit in various fractions of mouse brain and cross-reacts with the catalytic unit from lamb kidney, duck salt gland, and electroplax. The same polypeptide in brain and salt gland is recognized by antiserum raised against purified lamb kidney enzyme. Light microscopy was performed with the peroxidase-conjugated second antibody method. In mouse cerebellum, immunochemical staining outlines Purkinje cell and granule cell perikarya. Intense activity is associated with regions of high synaptic content including the pericellular basket meshes and preaxonal regions of Purkinje cells and the glomeruli in the granular layer. In the molecular layer, the neuropil is diffusely reactive with distinct vertically oriented processes evident. White matter exhibits light stain deposition. Choroid plexus presents abundant reaction product only at ependymal apical surfaces, while the ependymal lining of the fourth ventricle displays little or no immunoreactivity. Specificity of the antiserum was demonstrated further in mouse kidney where staining conforms to the well-characterized localization of the enzyme along basolateral surfaces of cortical and medullary tubules. The biochemical and immunocytochemical data show the efficacy of generating antisera to brain (Na+ + K+)-ATPase using catalytic polypeptide as an immunogen.

Animals↗
PubMed1983

Influence of dixyrazine on intestinal and renal vasoconstrictor responses during fentanyl-nitrous oxide anaesthesia.

In cats (n = 24) anaesthetized with fentanyl-nitrous oxide and diazepam, stimulation of the hypothalamic defence-alarm area (DA) or afferent activation of somatic pain fibres (SA), elicited a pronounced increase in intestinal (DA 297%, SA 107%) and renal (DA 214%, SA 90%) vascular resistance as well as a decrease in diuresis. These stress-related responses were markedly counteracted by dixyrazine (0.15-0.5 mg X kg-1 b.w.i.v.), especially in the kidney where the subsequent increase in vascular resistance to DA and SA stimulations amounted to only 25% and 13%, respectively, while diuresis increased. Corresponding data for stimulation-induced increases in intestinal vascular resistance after dixyrazine were DA 156% and SA 28%. Dixyrazine is suggested to act both through interaction with peripheral alpha-adrenergic mechanisms in control of vascular tone and through central nervous cardiovascular reflex depression. In man (n = 7), during a similar form of anaesthesia, portal vein blood flow (1137 +/- 177 ml) was measured by the continuous thermodilution method. Preportal tissue vascular resistance during surgery decreased significantly (11.3 vs 8.7 kPa X min X ml-1 X 10(-3] after i.v. dixyrazine (0.15 mg X kg-1 b.w.). A concomitant increase in oxygen uptake in preportal tissues occurred (19.9 ml min-1 vs 24.5 ml X min-1).

Aged↗
PubMed1983

Portal blood flow in man during surgery, measured by a modification of the continuous thermodilution method.

The continuous thermodilution method was applied to measurement of peroperative portal venous blood flow in man in relation to hepatic surgery during steady-state enflurane nitrous oxide anaesthesia. Portal-vein catheterization through the umbilical vein for cytostatic administration and blood flow measurements was successful in 14 out of 17 patients with primary or secondary liver tumours. Mean portal blood flow was 1195 +/- 179 ml X min-1 (mean +/- SEM), corresponding to 17.6 +/- 2.9 ml X min-1 X kg-1 body weight. Mean oxygen consumption in the prehepatic splanchnic area was 0.27 +/- 0.04 ml X min-1 X kg-1. No significant complications were associated with the catheterization or flow recording procedures. The described method offers possibilities for haemodynamic and pharmacokinetic investigations in man.

Catheterization↗
PubMed1982

Clonal lethality caused by the yeast plasmid 2 mu DNA.

Strains of Saccharomyces that carry the nib allele of a nuclear gene exhibit a "nibbled" colony morphology if they also harbor the plasmid 2 mu DNA. I have found that the expression of the nibbled phenotype is correlated with the presence of a subpopulation of abnormally large cells that give rise to mortal clones. Large cells apparently become large as a consequence of a defect in DNA replication or nuclear division. Large nib cells contain twice as much 2 mu DNA per microgram of total DNA as small nib cells do, and elevated 2 mu DNA copy number is the cause, not the effect, of increased cell size. It appears that the NIB allele can prevent an increase in 2 mu DNA copy number, but cannot produce a decrease once the copy number has exceeded the normal level. I propose, therefore, that the NIB gene product normally represses the amplification of 2 mu DNA copy number, and that the nib allele is partially defective in this function.

Cell Cycle↗
PubMed1982

Morphological and electrophysiological mapping of giant neurons in the suboesophageal ganglia of Helix pomatia.

1. To reveal the morphology of the suboesophageal ganglia of Helix pomatia, the connective tissue was completely removed and the preparations stained whole-mount with methyl green-pyronin G to display the relative locations of the neurons. 2. Fifteen large cells which could be recognized as individuals in at least 75% of the preparations investigated, were morphologically identified by size, position and color. 3. The cells were electrophysiologically characterized with respect to spontaneous activity, synaptic input from peripheral nerves, and response to application of drugs (e.g. ACh, DA and 5-HT). 4. The peripheral axonal projections of eight of the major identified cells were investigated by intracellular CoCl2 injection and by cobalt backfilling of the peripheral nerves.

Animals↗