Studies on equine lipid metabolism. 2. Lipolytic activities of plasma and tissue lipases in large horses and ponies

The enzymatic fundamentals of lipid metabolism of equine have not been thoroughly investigated at this point in time. It is still unclear why ponies in contrast to horses may become hyperlipaemic when coming negative energy balance. In this study, the activities of the triglyceride-cleaving key enzymes of ponies are large bred horses were investigated in order to obtain insight into the aetiology of the syndrome. The objective of the study was to measure the activities of hormone-sensitive lipase (HSL), lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) in ponies and horses in ex vivo in vitro assays. Norepinephrine (NE) stimulated pony adipocytes to release FFA in a linear fashion (4.57 +/- 2.09 nmol FFA.10(5) cells-1.min-1). This was not observed in horses. Lipolysis was significantly higher in fat cells of ponies than in horses when adenosine deaminase (ADA) and NE were added (12.71 +/- 3.12 vs. 1.96 +/- 1.22 nmol FFA.10(5) cells-1.min-1). Relative inhibition of lipolysis by the action of insulin was comparable in adipocytes of horses and ponies. However, absolute FFA release in pony fat cells was as high as the maximal NE and ADA stimulated lipolysis in horse adipocytes. Postheparin plasma lipase activities in ponies and horses did not differ between the sub-species. This finding was supported by the results obtained from measurement of LPL activity in adipose and muscle tissue showing only a tendency of increased activities in pony explants when compared to horse tissue incubations. This study further supports the hypothesis that differences in regulation of TG release from fat stores rather than clearance of TG from plasma is causative for the development of hyperlipaemia in ponies. Abbreviations used: ADA, adenosine deaminase; BW, body weight; FFA, free fatty acid; HSL, hormone-sensitive lipase; HTGL, hepatic triglyceride lipase; LPL, lipoprotein lipase; NE, norepinephrine; SDS, sodium dodecyl sulfate; TG, triglyceride; VLDL, very low density lipoprotein.     

Pathobiochemical importance of phospholipases for the release of mast cell mediators

We investigated the influence of polyunsaturated fatty acids on the activity of the cytosolic phospholipase A2 (cPLA2) in the canine mastocytoma cell line C2 as a model for canine atopic dermatitis (CAD). Cells were cultured in a basic medium or in media supplemented with different fatty acids (14 microM) for eight days. The supplemented fatty acids were linoleic acid (18:2n6), alpha-linolenic acid (18:3n3), gamma-linolenic acid (18:3n6) and docosahexaenoic acid (22:6n3). We measured enriched concentrations of the added fatty acid, their delta6-desaturated and elongated products. However, delta5-desaturated products were not increased. Culturing of C2 in 18:3n3 supplemented medium reduced the cPLA2 activity. Furthermore in these cells and in C2 cultured in 22:6n3 supplemented medium decreased the cPLA2 activity after stimulation. The reduced cPLA2 activity by the changed fatty acid pattern of C2 cultured in 18:3n3 or 22:6n3 possibly explain the beneficial effects of these fatty acids in CAD because increased cPLA2 activity is accompanied by enhanced release of proinflammatory type 2 prostaglandins and type 4 leukotrienes.     

The evaluation of hormone and metabolite profiles after the infusion of glucose, propionate and butyrate in cattle

Lactational performance in high-yielding dairy cows has its limits in metabolic processes. Energy metabolism is maintained by mobilization of body reserves via hormonal regulation, resulting in lipolysis and sometimes ketoacidosis. For characterization of such conditions the intraruminal and intravascular application of glucose and volatile fatty acids was used in ruminant physiology. On the other hand the results of such experiments were correlated to actual and potential milk yield. For this investigation pairs of monozygous Holstein Friesian twins were tested as heifers and as cows by intravenous infusion of glucose, propionate and butyrate after 18 hours of feed withdrawal. Insulin, growth hormone, glucose, free fatty acids and beta-hydroxybutyrate were measured before, during and up to 4 hours after infusion of substrates. Each substrate caused a transient change in plasma concentrations of metabolites and hormones. Differences between heifers and cows are discussed with the time series of the mean concentrations. The reaction profile of each animal was characterized by different parameters. Analysis of these parameters revealed a close relation between hormones and metabolites even under the conditions of the load-test.     

Die Wirkung einer Vitamin-A-Zulage auf die Konzentration einzelner antioxidativer Komponenten und die Antioxidative Kapazität im Serum von Ponys

Einleitung
Das Plasma enthält wenige antioxidative Enzyme, ist aber reich an anderen hoch- und niedermolekularen antioxidativen Molekülen. Hier sind die Plasma-Proteine mit ihren Thiolgruppen, allen voran Albumin, zu nennen. Von den niedermolekularen Verbindungen haben Urat, Ascorbat, Thiole, Bilirubin und α-Tocopherol eine Bedeutung. Als Parameter für die oxidative Homöostase des Organismus kann die Bestimmung der antioxidativen Kapazität des Plasmas und anderer biologischer Flüssigkeiten herangezogen werden (L ewin and P opov 1994). In den letzten Jahren wurde eine Vielzahl von Methoden zur Bestimmung der Antioxidativen Kapazität von Einzelsubstanzen und biologischem Material entwickelt. Die Generation freier Radikale geschah photochemisch (P opov and L ewin 1994), chemisch oder chemisch-enzymatisch (P ascual and R omay 1992). Die antioxidative Wirkung wurde dann über Radikal-Spintrapping (S oriani et al. 1994), Fluoreszens (R-Phycoerythrin) oder Chemilumineszenz (Luminol) ermittelt.
Die genannten Verfahren haben bisher kaum Eingang in die Tiermedizin gefunden, obwohl eine Reihe metabolischer Dysfunktionen bei Haustieren mit Störungen des peroxidativen Stoffwechsels in Zusammenhang gebracht werden (F uhrmann et al. 1996; M udron et al. 1997; S allmann 1998).
Ein weiterer Aspekt der Antioxidativen Kapazität ist die bei einigen Spezies beschriebene verminderte Vitamin E-Retention bei hohen Gaben von Vitamin A (Huhn: C ombs 1976; F rigg and B roz 1983; Ratte: J enkins and M itchell 1986; Schwein: H oppe and S ch & ouml ; ner 1993). Demzufolge erhob sich die Frage, ob auch beim Pony eine überhöhte Gabe von Vitamin A die Tocopherolgehalte und damit den antioxidativen Stoffwechsel negativ beeinflußt. Dazu wurden neben einzelnen antioxidativen Komponenten die Antioxidative Kapazität des Serums mit zwei unterschiedlichen Methoden bestimmt.     

Investigations on manifestations of vitamin E and selenium deficiency in sheep and goats]

In a prospective study from 1991-1998 plasma concentrations of alpha-Tocopherole (VitE) and Selenium (Se) were analysed in 125 sheep and 32 goats with generalised motor disturbances or elevated plasma-activities of Creatine-Kinase (CK). VitE-values < 1.0 mg/l and Se-values < 0.08 mg/l were regarded as deficiency. Diagnosis of pathological manifestations was based on blood enzyme values. CK-values in plasma > 300 U/l or Aspatate-Amino-Transferase (ASAT) > 150 U/l were regarded as myopathy, Glutamat-Dehydrogenase-values > 25 U/l indicated a hepatopathy. 92 (74%) sheep and 10 (31%) goats showed VitE and/or Se deficiency. There were no principal differences in the frequencies of myopathies (67%) and hepatopathies (46%) between pure VitE deficiencies or pure Se deficiencies or combinations of both deficiencies. In VitE deficiency and in the combinations of VitE and Se deficiency the elevations of the enzyme activities were more pronounced than in Se deficiency. The lethality was significantly higher in combined VitE + Se deficiencies (73%) than in pure VitE or Se deficiencies (35%). Combined VitE + Se deficiencies occurred more frequent in lambs (53%) and was often accompanied by anaemia (29%) and hypoproteinaemia (32%). The diagnostic reliability of ASAT and GLDH for the detection of VitE and Se deficiency was better than CK. Diagnostic and therapeutic strategies in flock diseases are discussed.