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Biomedical subjects

J Knudsen

Publications and source records attributed to J Knudsen.

At least 109 records · Page 6Linked to original sources
PubMed1987

Effect of exogenous long-chain fatty acids on lipid biosynthesis in dispersed ruminant mammary gland epithelial cells: esterification of long-chain exogenous fatty acids.

Dispersed epithelial cells from lactating bovine and goat mammary glands incorporated acetate into all fatty acids (C4 to C16) that were incorporated into mainly triacylglycerols. The cells secreted free fatty acids only into the incubation medium, and this secretion was dependent on the concentration of albumin and the type and amount of exogenous fatty acid added to the medium. Addition of palmitic acid to the incubation medium stimulated synthesis and incorporation of fatty acids synthesized de novo into triacylglycerols, whereas stearic and linoleic acid were inhibitory.

Acetates↗
PubMed1987

Effect of exogenous long-chain fatty acids on individual fatty acid synthesis by dispersed ruminant mammary gland cells.

We investigated the effect of exogenously added fatty acids on de novo synthesis of individual fatty acids and their incorporation into triacylglycerols by dispersed lactating ruminant mammary gland epithelial cells. Palmitate addition strongly stimulated synthesis and incorporation of butyrate and, to a smaller extent, palmitate synthesis and incorporation. Oleic acid strongly inhibited synthesis of all fatty acids except butyrate, whereas the effect of lauric acid was nearly neutral. Free fatty acid depletion of the mammary gland cells potentiated the effect of palmitate and made oleate less inhibitory.

Animals↗
PubMed1986

Synthesis of milk specific fatty acids and proteins by dispersed goat mammary-gland epithelial cells.

The method now described for preparation of dispersed lactating goat mammary-gland cells gives a high yield of morphologically and functionally normal mammary cells. The cells synthesize specific goat milk fatty acids in the right proportions, and they respond to hormones by increased protein synthesis. The cells can be frozen and thawed without losing the above properties, which makes them an excellent tool for metabolic and hormonal studies.

Acetates↗
PubMed1986

Absence of monoacylglycerol pathway for triacylglycerol synthesis in goat mammary gland.

Both subcellular fractions of lactating goat mammary gland and dispersed lactating goat mammary-gland cells were able to incorporate [U-3H]2-O-hexadecylglycerol into monoalkyl-monoacylglycerol. However, the incorporation of [U-3H]2-O-hexadecylglycerol into monoalkyl-monoacylglycerol by dispersed cells was not accompanied by incorporation of fatty acid synthesized de novo or added radiolabelled fatty acid. The result therefore shows that an active monoacylglycerol pathway does not exist in goat mammary gland.

Acetates↗
PubMed1986

Parallel measurements of heat production and thermogenin content in brown fat cells during cold acclimation of rats.

The maximum thermogenic capacity of brown fat cells from control and cold acclimated rats was measured using a continuous-flow microcalorimetric system. The content of the 32.000 D, brown fat specific protein, thermogenin, was measured in the cells used for heat production measurements by competitive ELISA. The ratio between the maximal thermogenic capacity and the amount of thermogenin for control and cold acclimated rats was compared. It was found that the ratio between the two parameters decreased during cold acclimation due to a decrease in maximal thermogenic capacity and an increase in the amount of thermogenin, indicating regulation of heat production either at thermogenin or receptor level.

Acclimatization↗
PubMed1986

Echinostoma revolutum: resistance to secondary and superimposed infections in mice.

A complete or almost complete resistance (94-100%) to a superimposed Echinostoma revolutum infection existed in mice harboring 20-, 30-, and 40-day-old infections in the range of 2-4 to 30-35 worms, but no resistance was found at challenge Day 10. A similar high level of resistance (85-100%) also existed in mice for at least 6 weeks after natural expulsion of a primary 6 metacercarial infection and for at least 5 weeks after anthelmintic termination of a 30-day-old 20 metacercarial infection. Thymus-deficient nude mice failed to develop resistance to a superimposed infection, and the resistance in normal mice was inhibited by corticosteroid treatment. These findings are all in favor of a host immune response being responsible for the resistance against both a secondary and a superimposed infection. Nearly all the worms of a superimposed infection were, in resistant mice, expelled prior to 24 hr following infection (rapid expulsion), and the few worms circumventing this early expulsion persisted for at least 8 days. Newly excysted juvenile worms implanted intraduodenally into resistant mice were rejected to the same degree as juvenile worms from an oral metacercarial infection indicating that the newly excysted juvenile worms are the target of the host immune response. However, 7-day-old worms implanted intraduodenally into resistant mice survived indicating that adaptation to the host immune response had occurred. In conclusion, this host-parasite model is an example of concomitant immunity because the immunological mechanism responsible for the expulsion of the superimposed infection had no effect on the number of primary worms present.

Adrenal Cortex Hormones↗
PubMed1986

Test-retest reliability of the Minnesota Leisure Time Physical Activity Questionnaire.

The test-retest reliability of the Minnesota Leisure Time Physical Activity Questionnaire was assessed by two administrations of the questionnaire at a five-week interval in 140 adults from a general population sample and at a two-week interval in 150 male participants in the Multiple Risk Factor Intervention Trial (MRFIT). In both study populations, there was a slight, nonsignificant drop in reported leisure time energy expenditure between the test and the retest. The largest test-retest differences in estimated energy expenditure were found among those reporting greater levels of energy expenditure. Nevertheless, in both study populations Spearman rank correlation coefficients between the test and retest were high--0.79 to 0.88 for total activity and ranging 0.69-0.86 among the light, moderate, and heavy intensity subcategories. The high reliability of the Minnesota Leisure Time Physical Activity Questionnaire corroborates its utility in epidemiologic research.

Adult↗
PubMed1986

Epidemiological studies on Schistosoma bovis in Iringa Region, Tanzania.

Various aspects of the epidemiology of Schistosoma bovis were studied over a one-year period in Iringa Region, Tanzania. An abattoir survey revealed an overall prevalence rate of 30.8% in cattle and 3.8% in goats in the area, and field studies on two dairy farms both providing good opportunities for schistosome transmission provided information concerning the transmission ecology of S. bovis in relation to different types of grazing and water supply. The traditional management system on one farm with a large number of cattle utilizing a limited water resource highly suitable for sustaining populations of the snail host Bulinus africanus resulted in intensive transmission as evidenced by uptake of massive infections in calves and development of resistance to S. bovis challenge in dairy cows. On another farm, appropriate management comprising watering of cattle at a B. africanus-free pond provided the background for less intensive transmission in that transmission risk was confined to occasional contact with water contact sites of secondary importance. Besides, the transmission pattern as regards intensity and seasonality was affected markedly by the geographical and seasonal distribution of the host snail B. africanus. Thus, transmission in canals and temporary ponds was limited mainly to the dry season and the end of the rainy season, respectively, while transmission in permanent ponds occurred intermittently throughout at least most of the year. It is concluded that prevention of severe loss of productivity in domestic ruminants due to schistosome infections should be possible using strategic management procedures provided that essential information is available concerning the pattern of transmission in the particular area.

Abattoirs↗
PubMed1985

Mammalian fatty acid synthetase is a structurally and functionally symmetrical dimer.

We have explored a comprehensive experimental approach to determine whether the two condensing-enzyme active centers of the mammalian fatty acid synthetase are simultaneously functional. Our strategy involved utilization of trypsinized fatty acid synthetase, which is a nicked homodimer composed of two pairs of 125 + 95-kDa polypeptides. These core polypeptides lack the chain-terminating thioesterase domains but retain all other functional domains of the native enzyme and can assemble long-chain acyl moieties at a rate equal to that of the native enzyme. The 4'-phosphopantetheine content of these enzyme preparations, estimated from the amount of beta-alanine present, from the amount of taurine formed by performic acid oxidation and from the amount of carboxymethylcysteamine formed by alkylation with iodo[2-14C]acetate, was typically 0.86 mol/mol 95-kDa polypeptide. The stoichiometry of long-chain acyl-enzyme synthesis, measured with radiolabeled precursors, indicated that 0.84 mol acyl-chains were assembled/mol 95-kDa polypeptide. When the small amount of apoenzyme present is taken into account, this stoichiometry translates to 1.94 acyl chains per holoenzyme dimer. The 125-kDa polypeptide of one subunit could be cross-linked to the 95-kDa polypeptide of the other subunit by 1,3-dibromo-2-propanone yielding a single molecular species of 220 kDa. Cross-linking was accompanied by a loss of condensing-enzyme activity. This result is consistent with a structurally symmetrical model for the animal fatty acid synthetase [J.K. Stoops and S.J. Wakil (1981) J. Biol. Chem. 256, 5128-5133] in which the juxtaposed 4'-phosphopantetheine and cysteine thiols of opposing subunits that form the two potential catalytic centers for condensing activity are readily susceptible to cross-linking. Both half-maximal cross-linking and 50% inhibition of activity were observed with 1 mol 1,3-dibromo-2-propanone bound/mol enzyme. After assembly of long-chain acyl moieties on the 4'-phosphopantetheine residues, no vacant condensing-enzyme active sites were demonstrable either by cross-linking with 1,3-dibromo-2-propanone or by formation of carboxymethylcysteamine on treatment with iodoacetate. These results are consistent with a structurally and functionally symmetrical model for the mammalian fatty acid synthetase in which the two condensation sites are simultaneously active.

Acetone↗
PubMed1985

Stoichiometry of substrate binding to rat liver fatty acid synthetase.

Two rat liver fatty acid synthetase preparations, containing 1.6 and 2.0 mol of 4'-phosphopantetheine/mol of synthetase, showed specific activity of 2006 and 2140 nmol of NADPH oxidized/min per mg of protein respectively. The two synthetase preparations could be loaded with either 3.3-4.4 mol of [1-14] acetate or 2.9-3.7 mol of [2-14C]malonate, by incubation with either [1-14C] acetyl-CoA or [2-14C]malonyl-CoA. The 4'-phosphopantetheine site could be more than 90% saturated and the serine site about 80% saturated with malonate derived from malonyl-CoA. However, with acetyl-CoA as substrate, binding at both the 4'-phosphopantetheine and cysteine thiol sites did not reach saturation. We interpret these results to indicate that, whereas the equilibrium constant for transfer of substrates between the serine loading site and the 4'-phosphopantetheine site is close to unity, that for transfer of acetyl moieties between the 4'-phosphopantetheine and cysteine sites favours formation of the 4'-phosphopantetheine thioester. Thus, despite the apparent sub-stoichiometric binding of acetate, the results are consistent with a functionally symmetrical model for the fatty acid synthetase which permits simultaneous substrate binding at two separate active centres.

Acetates↗
PubMed1985

Evidence that the medium-chain acyltransferase of lactating-goat mammary-gland fatty acid synthetase is identical with the acetyl/malonyltransferase.

Competitive binding experiments with malonyl-CoA and [1-14C]acetyl-CoA, [1-14C]butyryl-CoA or [1-14C]decanoyl-CoA indicate that all these substrates are transferred to lactating-goat mammary-gland fatty acid synthetase by the same transferase. Isolation and determination of the amino acid sequence of [1-14C]decanoyl-labelled CNBr-cleavage peptide from the decanoyltransferase site showed that this transferase is identical with the acetyl/malonyltransferase.

Acetyl Coenzyme A↗
PubMed1985

Amino acid sequence around the active-site serine residue in the acyltransferase domain of goat mammary fatty acid synthetase.

Goat mammary fatty acid synthetase was labelled in the acyltransferase domain by formation of O-ester intermediates by incubation with [1-14C]acetyl-CoA and [2-14C]malonyl-CoA. Tryptic-digest and CNBr-cleavage peptides were isolated and purified by high-performance reverse-phase and ion-exchange liquid chromatography. The sequences of the malonyl- and acetyl-labelled peptides were shown to be identical. The results confirm the hypothesis that both acetyl and malonyl groups are transferred to the mammalian fatty acid synthetase complex by the same transferase. The sequence is compared with those of other fatty acid synthetase transferases.

Acetyl Coenzyme A↗
PubMed1985

Limitations in the use of the enzyme-linked immunosorbent assay (ELISA) for identification and quantification of thermogenin.

The use of immunological assays, ELISA and RIA, for the identification and quantification of thermogenin (the brown adipose tissue-specific, GDP-binding, 32 kDa uncoupling protein) raises doubts regarding the exclusive occurrence of thermogenin in brown adipose tissue. Weak reactions between mitochondria from rat liver, rat skeletal and heart muscle and hamster white adipose and thermogenin antibodies have been observed (Cannon et al., 1982; Lean et al., 1983; Hansen et al., 1984). In order to study whether these reactions were due to thermogenin in tissues other than brown adipose tissue (BAT) or due to non-specific binding of thermogenin antibodies, a protein from rat liver mitochondria and a protein from tubifex mitochondria were isolated by the same procedure as thermogenin. The 2 proteins had almost the same molecular weight as thermogenin and reacted with thermogenin antibodies in ELISA and dot-blotting, but did not bind GDP and had an amino acid composition different from that of thermogenin. It is concluded that the weak reactions seen between thermogenin antibodies and mitochondria from different tissues other than BAT are due to non-specific binding, and that antibody cross-reactivity alone is unsuitable for the identification of thermogenin.

Adipose Tissue↗
PubMed1984

Triacylglycerol synthesis in goat mammary gland. The effect of ATP, Mg2+ and glycerol 3-phosphate on the esterification of fatty acids synthesized de novo.

Goat mammary-gland microsomal fraction by itself induces synthesis of medium-chain-length fatty acids by goat mammary fatty acid synthetase and incorporates short- and medium-chain fatty acids into triacylglycerol. Addition of ATP in the absence or presence of Mg2+ totally inhibits triacylglycerol synthesis from short- and medium-chain fatty acids, and severely inhibits synthesis de novo of medium-chain fatty acids. The inhibition by ATP of fatty acid synthesis and triacylglycerol synthesis de novo can be relieved by glycerol 3-phosphate. The effect of ATP could not be mimicked by the non-hydrolysable ATP analogue, adenosine 5'-[beta,gamma-methylene]triphosphate and could not be shown to be caused by inhibition of the diacylglycerol acyltransferase by a phosphorylation reaction. Possible explanations for the mechanism of the inhibition by ATP are discussed, and a hypothetical model for its action is outlined.

Adenosine Triphosphate↗