RESUMO
Although it is well recognized that dietary supplementation with fish oil improves clinical symptoms in dogs suffering from osteoarthritis, the molecular basis for the dietary benefit is not yet completely resolved in dogs. This study was designed to further clarify how polyunsaturated fatty acids (PUFA) affect key factors of cartilage degeneration in a canine cell culture system mimicking osteoarthritis. Canine chondrocytes were incubated either without or with 10 µm of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), arachidonic acid (AA) or 3.6 µm ibuprofen (Ibu) as positive control for 6 days. After the supplementation, cells were stimulated with 10 ng/ml interleukin-1ß (IL-1ß) for another 48 hr to induce osteoarthritic changes, or left unstimulated. We analysed fatty acid uptake via gas-liquid chromatography, nitric oxide (NO) production via Griess assay, prostaglandin E (PGE) production via ELISA and relative gene expression of several cartilage matrix proteinases, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 via RT-qPCR. After supplementation, the chondrocytes rapidly incorporated the PUFA into their fatty acid pools. The stimulation with IL-1ß caused a marked increase of most of the inflammatory markers measured. N-3 PUFA EPA reduced IL-induced gene expression of iNOS and corresponding production of NO. N-6 PUFA AA also decreased iNOS and NO, but furthermore lowered gene expression of matrix metalloproteinase-3. On the other hand, AA upregulated the aggrecanase ADAMTS-5 and augmented the release of PGE. The effect of n-3 PUFA DHA turned out to be negligible. Our results reveal molecular mechanisms by which PUFA affect degenerative joint disease in dogs. Of particular importance is that not only EPA but also AA decreased several inflammatory markers in our model. Thus, we conclude that an appropriate balance of both n-3 and n-6 fatty acids deserves more attention in dietary interventions.
Assuntos
Ácido Araquidônico/farmacologia , Condrócitos/efeitos dos fármacos , Ácidos Docosa-Hexaenoicos/farmacologia , Ácido Eicosapentaenoico/farmacologia , Inflamação/veterinária , Osteoartrite/veterinária , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Anti-Inflamatórios não Esteroides/farmacologia , Ácido Araquidônico/administração & dosagem , Biomarcadores/metabolismo , Condrócitos/metabolismo , Ácidos Docosa-Hexaenoicos/administração & dosagem , Doenças do Cão/tratamento farmacológico , Doenças do Cão/metabolismo , Cães , Ácido Eicosapentaenoico/administração & dosagem , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Ibuprofeno/administração & dosagem , Ibuprofeno/farmacologia , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Osteoartrite/tratamento farmacológicoRESUMO
Conjugated linoleic acids (CLA), particularly cis-9,trans-11 (c9t11) and trans-10,cis-12 (t10c12), are used as feed additives to adapt to constantly increasing demands on the performance of lactating cows. Under these feeding conditions, the rumen wall, and the rumen epithelial cells (REC) in particular, are directly exposed to high amounts of CLA. This study determined the effect of CLA on the fatty acid (FA) metabolism of REC and expression of genes known to be modulated by FA. Cultured REC were incubated with c9t11, t10c12, and the structurally similar FA linoleic acid (LA), oleic acid (OA), and trans-vaccenic acid (TVA) for 48 h at a concentration of 100 µM. Cellular FA levels were determined by gas chromatography. Messenger RNA expression levels of stearoyl-CoA desaturase (SCD) and monocarboxylate transporter (MCT) 1 and 4 were quantified by reverse transcription-quantitative PCR. Fatty acid evaluation revealed significant effects of CLA, LA, OA, and TVA on the amount of FA metabolites of ß-oxidation and elongation and of metabolites related to desaturation by SCD. The observed changes in FA content point (among others) to the ability of REC to synthesize c9t11 from TVA endogenously. The mRNA expression levels of SCD identified a decrease after CLA, LA, OA, or TVA treatment. In line with the changes in mRNA expression, we found reduced amounts of C16:1n-7 cis-9 and C18:1n-9 cis-9, the main products of SCD. The expression of MCT1 mRNA increased after c9t11 and t10c12 treatment, and CLA c9t11 induced an upregulation of MCT4. Application of peroxisome proliferator-activated receptor (PPAR) α antagonist suggested that activation of PPARα is involved in the changes of MCT1, MCT4, and SCD mRNA expression induced by c9t11. Participation of PPARγ in the changes of MCT1 and SCD mRNA expression was shown by the application of the respective antagonist. The study demonstrates that exposure to CLA affects both FA metabolism and regulatory pathways within REC.
Assuntos
Ácidos Graxos/metabolismo , Ácidos Linoleicos Conjugados/farmacologia , Rúmen/metabolismo , Ovinos/metabolismo , Animais , Células Cultivadas , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Feminino , Lactação/efeitos dos fármacos , Transportadores de Ácidos Monocarboxílicos/genética , Ácidos Oleicos , PPAR alfa/fisiologia , PPAR gama/fisiologia , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Estearoil-CoA Dessaturase/genéticaRESUMO
Canine atopic dermatitis (CAD) is an inflammatory skin disorder that is characterized by pruritus and associated cutaneous changes. Treatment interventions include allergen avoidance, allergen-specific immunotherapy as well as a symptomatic therapy using glucocorticoids and antihistamines. In addition, a dietary intervention using polyunsaturated fatty acids (PUFA) has been shown to alleviate symptoms in some dogs. Although the beneficial effects of PUFA in the treatment of CAD have been known for several years, their mode of action remains unclear. This review discusses the evidential basis of the therapeutic use of dietary PUFA in the treatment of CAD. Particular emphasis will be placed on the role of cutaneous mast cells. In addition, recent evidence from in vitro studies on the regulation of mast cell exocytosis will be used to build a mechanistic model of the active principle of PUFA. It is proposed that dietary PUFA are integrated into mast cell membranes resulting in a reorganization of membrane microdomains. This may then be accompanied by functional changes of membrane-associated proteins such as the phospholipases D (PLD), enzymes having an important impact on mast cell exocytosis processes.
Assuntos
Dermatite Atópica/veterinária , Doenças do Cão/tratamento farmacológico , Ácidos Graxos Insaturados/uso terapêutico , Mastócitos/efeitos dos fármacos , Ração Animal , Animais , Dermatite Atópica/tratamento farmacológico , Suplementos Nutricionais , Cães , Mastócitos/metabolismoRESUMO
An elevated level of long-chain n-3 fatty acids (FA) in tissue membranes has a positive influence on the progression and treatment of many diseases. Therefore, dietary supplementation of n-3 FA is recommended in some diseases. Even though n-3 FA are absorbed readily from the diet, their incorporation into tissues may be compromised in diseased animals. In a clinical setting, it is desirable to monitor the success of dietary intervention. Plasma FA as well as erythrocyte membrane (EM) FA can be used to monitor dietary FA intake. This study compares FA from EM and plasma with regard to their reaction time and reliability for monitoring dietary changes of tissue FA profiles in dogs. Thirty dogs were divided into three groups and fed for 12 weeks. The control group (CONT) was fed a commercial standard diet low in n-3 FA. One group received the standard diet and 85 mg/kg body weight of a docosahexaenoic acid (DHA) concentrate (ADD). The third group was fed a commercial dog food containing fish oil (FO), which is rich in eicosapentaenoic acid (EPA). EM and plasma FA profiles were analysed by GC separately. Data on EM FA were published recently. n-3 FA in plasma reached the new level after 2 weeks (8 weeks in EM). Dietary differences between DHA and EPA are obvious after 1 week already. The concomitant decrease in plasma n-6 FA differed between ADD and FO. In general, the correlation of n-6 FA between plasma and EM was low. We therefore conclude that analysis of plasma FA is sufficient for monitoring a diet-induced increase in tissue n-3 FA in dogs. However, EM FA should be analysed if the effect of dietary intervention on tissue n-6 FA is important.
Assuntos
Ração Animal/análise , Dieta/veterinária , Cães/sangue , Eritrócitos/metabolismo , Ácidos Graxos/sangue , Ácidos Graxos/metabolismo , Animais , Gorduras na Dieta/análise , Gorduras na Dieta/farmacologia , Suplementos Nutricionais , Cães/metabolismo , Eritrócitos/química , Ácidos Graxos/químicaRESUMO
BACKGROUND: Studies in mammals proved dietary organic selenium (Se) being superior to inorganic Se regarding effects on growth performance, antioxidative status, immune response, and Se homeostasis. However, the picture of possible effects of different Se sources and - levels can be expanded. The present field study evaluated the effects on weight gain, hematological and selected biochemical variables as well as plasma concentrations of vitamin E (vitE), total Se and selenobiomolecules in piglets throughout the suckling period. METHODS: Piglets were monitored from birth to 38 days of age (d). The mother sows' diets were enriched with l-selenomethionine (SeMet-0.26 and -0.43 mg Se/kg feed) or sodium selenite (NaSe-0.40 and -0.60 mg Se/kg feed) from 1 month prior to farrowing until the end of lactation period. Piglets received pelleted feed supplemented with Se similarly to the sows' diets from one week of age. Selenite at 0.40 mg Se/kg (NaSe-0.40) represents a common Se source and -level in pig feed and served as control diet. RESULTS: From 24d, piglets in SeMet-groups had higher mean body weight (BW) compared with piglets from sows fed NaSe-0.40. Furthermore, from five-d and above, piglets from sows fed NaSe-0.60 had significantly higher BW than offspring from sows fed NaSe-0.40. Neonatal piglets in group SeMet-0.43 had significantly lower red blood cell counts (RBC), hemoglobin (Hgb) and hematocrit (Hct) concentrations compared with piglets from sows fed with NaSe-0.40. Neonatal and 5d-old piglets in group SeMet-0.26 showed higher gamma-glutamyl transferase activity than piglets in group NaSe-0.40. From five d and above, group NaSe-0.60 excelled with increased specific hematological variables culminating at age 38d with increased Hct, mean corpuscular volume (MCV), and MC hemoglobin (MCH) as well as increased activities of aspartate transaminase and lactate dehydrogenase compared with the other groups. Generally, offspring in the SeMet groups had higher total Se-concentrations in plasma than those from sows fed selenite, and showed a dose-response effect on plasma Se-concentrations. Furthermore, SeMet-fed piglets had higher plasma levels of the selenoproteins (Sel) glutathione peroxidase 3 (GPx3) and SelP as well as selenoalbumin. Plasma vitE levels were significantly negatively correlated with RBC throughout trial period. CONCLUSIONS: Maternal supplementation with SeMet during gestation influenced hematology and clinical biochemistry in neonatal piglets in a different way than in offspring from sows receiving selenite enriched diets. Growth performance was positively influenced by both dietary Se source and Se level. Higher plasma levels of GPx3 observed in piglets receiving SeMet probably improved the protection against birth or growth related oxidative stress. These might prime the piglets for demanding situations as indicated by higher weight gain in offspring from sows fed with SeMet-supplemented diets. Our results on some enzyme activities might indicate that piglets fed NaSe-0.60 had to cope with increased levels of oxidative stress compared with those originating from sows fed SeMet or lower dietary levels of selenite. We assume that combining inorganic and organic Se sources in complete feed for breeding sows might be beneficial fro reproduction and the offspring's performance.
Assuntos
Antioxidantes/metabolismo , Comportamento Alimentar , Selenometionina/farmacologia , Selenito de Sódio/farmacologia , Animais , Animais Recém-Nascidos , Peso Corporal/efeitos dos fármacos , Dieta/veterinária , Especificidade de Órgãos/efeitos dos fármacos , Selênio/sangue , Suínos/sangue , Vitamina E/sangueRESUMO
BACKGROUND: Although the risk of developing dysglycaemia has been investigated in different communities this incidence is poorly studied in patients on maintenance haemodialysis (MHD). MATERIALS AND METHODS: In a multicentre observational cohort study the occurrence of dysglycaemia was assessed in 239 primary normoglycaemic end stage renal disease (ERSD) patients on MHD. Dysglycaemia (fasting blood glucose > 110 mg dL(-1), > 140 mg dL(-1) 2 h after food intake) or diabetes (fasting blood glucose > 126 mg dL(-1) or > 200 mg dL(-1) at any time) were defined according to WHO criteria and cases were compared with age matched controls within the cohort. RESULTS: Dysglycaemia was found in 82 primary normoglycaemic ESRD patients (34%) within 31 months after initiation of MHD. In 31 of these patients type 2 diabetes was diagnosed. When compared with matched control MHD patients differences in body mass index (BMI), HbA1c and postprandial blood glucose were detectable (P < 0.05). Increments in 0.1% of HbA1c were related with 11% higher odds for dysglycaemia (P = 0.002). In a subgroup of 36 primary normoglycaemic MHD patients who developed dysglycaemia event-free survival was 64%, 53%, 31%, 17% and 11% after 1, 2, 3, 4 and 5 years of haemodialysis treatment. CONCLUSION: Onset of dysglycaemia or diabetes is frequent in ESRD patients after onset of chronic haemodialysis. Routine measurement of blood glucose before and after haemodialysis should be implemented as a standard of care during MHD.
Assuntos
Diabetes Mellitus/etiologia , Falência Renal Crônica/complicações , Diálise Renal , Idoso , Biomarcadores/sangue , Diabetes Mellitus/sangue , Diabetes Mellitus/mortalidade , Progressão da Doença , Intervalo Livre de Doença , Feminino , Hemoglobinas Glicadas/análise , Humanos , Hiperglicemia/sangue , Hiperglicemia/etiologia , Falência Renal Crônica/mortalidade , Falência Renal Crônica/terapia , Masculino , Pessoa de Meia-Idade , Período Pós-Prandial , Estatísticas não ParamétricasRESUMO
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.
Assuntos
Dermatite Atópica/veterinária , Doenças do Cão/patologia , Ácidos Graxos Insaturados/farmacologia , Mediadores da Inflamação/metabolismo , Mastócitos/metabolismo , Fosfolipases/metabolismo , Animais , Linhagem Celular Tumoral , Dermatite Atópica/patologia , Modelos Animais de Doenças , Cães , Mastocitoma/patologia , Mastocitoma/veterinária , Fosfolipases/efeitos dos fármacosRESUMO
5-Monodeiodination of T4 and T3 and 5'-monodeiodination of T4 and rT3 were studied in brain homogenates of male Sprague-Dawley rats, aged 1-60 days. Portions of the homogenates were incubated with the substrates at 37 C for 30 min. The reaction products were estimated by specific RIAs. All of the four reactions were dependent upon time, temperature, pH, and upon the concentrations of substrate, thiol, and tissue protein. Maximal reactions were obtained between 40 and 160 mM dithioerythritol. T4 5'-deiodination proceeded optimally at pH 7.4 and 0.4 microM substrate, the other reactions at pH 8.5 and 10 microM substrate. The four reactions were inactivated by heat (56 C, 30 min) and inhibited by 10(-5) M iopanoic acid. Only rT3 5'-deiodination was inhibited by 3 X 10(-4) M propylthiouracil (greater than 95%). In cerebellum, basal ganglia, brainstem, and hypothalamus both T4 and T3 5-deiodinase activity were very high in perinatal rats [up to 5.56 pmol/(min X mg protein) in hypothalamus], and decreased rapidly with age. In cortex and olfactory bulb these enzyme activities were low after birth, followed by an increase during the growth spurt [up to 632 fmol/(min X mg protein) in olfactory bulb]. T4 and rT3 5'-deiodinase activity in all brain regions studied were at their lowest in perinatal rats. During and after the growth spurt an increase was observed [up to 457 fmol/(min X mg protein) in cerebellum]. The reciprocal course of 5- and 5'-deiodination between birth and growth spurt in most of the brain regions studied might lead to a reduced intracellular thyromimetic activity during the perinatal period.
Assuntos
Encéfalo/enzimologia , Iodeto Peroxidase/metabolismo , Animais , Animais Recém-Nascidos/metabolismo , Encéfalo/efeitos dos fármacos , Encéfalo/crescimento & desenvolvimento , Ditioeritritol/farmacologia , Concentração de Íons de Hidrogênio , Ácido Iopanoico/farmacologia , Masculino , Adeno-Hipófise/enzimologia , Propiltiouracila/farmacologia , Radioimunoensaio , Ratos , Ratos Endogâmicos , Temperatura , Tiroxina/metabolismo , Fatores de Tempo , Distribuição Tecidual , Tri-Iodotironina/metabolismo , Tri-Iodotironina Reversa/metabolismoRESUMO
The objective of this study was to investigate the effects of alpha-linolenic acid (18:3n-3) and linoleic acid (18:2n-6) on the fatty acid composition and the activity and release of mast cell mediators in the canine mastocytoma cell line C2. Cells were cultured in Dulbecco's modified Eagle's medium mixed with 50% Ham's F12 (containing linoleic acid 0.14 micro M). The basic medium (DEH) was supplemented with 0.14 micro M alpha-linolenic acid. 14.0 micro M alpha-linolenic acid (DEH-n-3) or 14.0 micro M linoleic acid (DEH-n-6) was added. Eight days after culturing of C2 in DEH-n-3 we measured elevated levels of n-3 fatty acids up to 22:3. The tryptase activity and the stimulated PGE2 production and histamine release were reduced. In contrast, after culturing of C2 in DEH-n-6 we determined elevated levels of n-6 fatty acids up to 20:3, increased tryptase activity and stimulated histamine release. Thus 18:3n-3 has anti-inflammatory effects in cultured canine mastocytoma cells.
Assuntos
Ácidos Graxos Essenciais/farmacologia , Mastócitos/efeitos dos fármacos , Animais , Linhagem Celular Tumoral , Quimases , Dinoprostona/biossíntese , Relação Dose-Resposta a Droga , Liberação de Histamina/efeitos dos fármacos , Ácido Linoleico/farmacologia , Mastócitos/metabolismo , Serina Endopeptidases/metabolismo , Fatores de Tempo , Triptases , Ácido alfa-Linolênico/farmacologiaRESUMO
Thyronine (T4), triiodothyronine (T3), and reverse-triiodothyronine (rT3) levels were evaluated in cerebrospinal fluid (CSF) and in serum of 12 patients with definite amyotrophic lateral sclerosis (ALS) by specific radioimmunoassays. Circulating microsomal and thyroglobulin antibodies were also evaluated. In all patients serum levels of T4, T3 and rT3 were within normal limits. In CSF, the rT3 levels were significantly elevated to 0.118 micrograms/l (mean), the T4 levels were not significantly elevated, and the T3 levels were below the detection limit of 0.03 micrograms/l. A correlation between the elevated rT3 levels in CSF and the severity or type of ALS could not be demonstrated by this study. The antithyroid antibodies (thyroglobulin antibodies, microsomal antibodies) showed normal titres and did not suggest disturbances of thyroid autoimmunity in the patients with ALS.
Assuntos
Esclerose Lateral Amiotrófica/sangue , Tironinas/sangue , Tri-Iodotironina Reversa/sangue , Tri-Iodotironina/sangue , Adulto , Idoso , Esclerose Lateral Amiotrófica/líquido cefalorraquidiano , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Tironinas/líquido cefalorraquidiano , Tri-Iodotironina/líquido cefalorraquidiano , Tri-Iodotironina Reversa/líquido cefalorraquidianoRESUMO
An important part in the use of genomic DNA libraries is the sequencing of identified clones for detailed information. In this study, methods for DNA sequence analysis were elaborated and employed for the k-casein gene, a bovine milk protein. The results encourage further research.
Assuntos
Bovinos/genética , DNA/química , Biblioteca Gênica , Animais , Clonagem MolecularRESUMO
Nutritional encephalomalacia (NE) in broiler chicken is considered as a peroxidative dysfunction caused by vitamin E-deficient diets. A feeding experiment was performed to investigate the consequences of feeding different fats in combination with increasing amounts of vitamin E on liver lipid peroxidation and plasma prostanoid pattern. Newly hatched chicks from hens on a vitamin E-poor diet were fed with either mainly linolenic, linoleic or oleic acid-rich oils in a vitamin E-deficient (5 ppm) basic diet. The animals were supplemented with vitamin E on three levels (0, 20 or 120 ppm). On appearance of the first symptoms of NE after 8 days post-hatching, the animals were examined. Typical symptoms with a high incidence only occurred in the group fed linoleic acid and 5 ppm vitamin E. Plasma prostanoids and microsomal alkane production in liver as a measure of endogenous lipid peroxidation were determined. The dietary conditions affected plasma prostaglandin E2 and thromboxane A2, but not prostacyclin. However, it seems unlikely that the prostanoids are involved in the pathogenesis of NE. Liver lipid peroxidation increased in vitamin E deficiency. The level of alkanes depended on the type of fat supplied. The consequences of the different dietary fats in combination with vitamin E deficiency on peroxidative metabolism of broiler chickens are evident, indicating that a high level of oxidative stress is imposed by the linoleic acid-rich fat.
Assuntos
Galinhas/metabolismo , Gorduras na Dieta/administração & dosagem , Encefalomalacia/veterinária , Peroxidação de Lipídeos , Doenças das Aves Domésticas/metabolismo , Vitamina E/administração & dosagem , Alcanos/metabolismo , Animais , Gorduras na Dieta/farmacologia , Encefalomalacia/etiologia , Encefalomalacia/metabolismo , Ácido Linoleico , Ácidos Linoleicos/administração & dosagem , Ácidos Linoleicos/farmacologia , Microssomos Hepáticos/metabolismo , Ácido Oleico , Ácidos Oleicos/administração & dosagem , Ácidos Oleicos/farmacologia , Doenças das Aves Domésticas/etiologia , Prostaglandinas/sangue , Vitamina E/farmacologia , Deficiência de Vitamina E/complicações , Deficiência de Vitamina E/veterinária , Ácido alfa-Linolênico/administração & dosagem , Ácido alfa-Linolênico/farmacologiaRESUMO
This study examined the shelf life of cured sausage under different packaging conditions from vitamin E supplemented pigs. One group (n=6) of crossbred pigs received a normal fattening diet containing 20 ppm α-tocopherol for 39 days before slaughter. Another group was fed a diet containing 410 ppm α-tocopherol during the same period. After slaughter, cured sausages were produced, packaged under three different atmospheric conditions, ripened for 4 weeks and then stored for 8 weeks (9 °C; 200 lux). The α-tocopherol content was recorded in the fat, liver, muscles and the sausage. TBARS, L*, a*, b* values and antioxidative capacity were evaluated in the sausage during storage. The results showed a transfer of vitamin E into tissues and sausage but no detectable effect on TBARS and colour stability. However, antioxidative effects of vitamin E were seen by provocation. Probably the effect of vitamin E in the sausage was masked by nitrite in the curing salt. The oxygen content of the packs had an influence on TBARS and colour stability.
RESUMO
This study investigated the inhibitory efficiency of all-rac-alpha-tocopherol, 2,6-ditert-butyl-p-cresole (BHT), and 6-hydroxy-2,5,7,8-tetramethyl-chroman-2-carbonic acid (Trolox) on determination of thiobarbituric acid-reactive substances (TBARS) and short-chain alkenals in rat liver homogenates. The concentration of TBARS was measured fluorophotometrically. Aldehydes were determined after derivatization with methylhydrazine by gas chromatography (GLC). The concentrations of alkenals and TBARS in liver homogenates were diminished when antioxidants were present during the sample preparation. It is suggested that in the absence of antioxidants the samples are autoxidized further during the preparative procedures. For the aldehyde determination all-rac-alpha-tocopherol was the most effective antioxidant to reduce the bias due to autoxidation, whereas for TBARS it was Trolox.
Assuntos
Aldeídos/análise , Antioxidantes/farmacologia , Fígado/química , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Aldeídos/metabolismo , Animais , Hidroxitolueno Butilado/farmacologia , Cromanos/farmacologia , Cromatografia Gasosa , Feminino , Fluorofotometria , Fígado/efeitos dos fármacos , Fígado/metabolismo , Oxirredução , Ratos , Ratos Wistar , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Vitamina E/farmacologiaRESUMO
The concentrations of hormones, metabolites and electrolytes in plasma of 45 ewes have been examined repeatedly during gestation (day 80 to 149) and at the third day of lactation. Healthy single pregnant (n = 8) and twin pregnant ewes (n = 12) have been compared with ewes sick with subclinical (3-hydroxybutyrate > 1,0 mmol/l, n = 6) and clinical (3-hydroxybutyrate > 1,6 mmol/l, n = 19) pregnancy ketosis. The concentration of 3-hydroxybutyrate, glucose, calcium, sodium and growth hormone in the plasma of the healthy animals were higher post partum than ante partum, while the concentrations of inorganic phosphorus, cortisol and total oestrogen were lower post partum than during pregnancy. In the pregnant ewes the concentrations of 3-hydroxybutyrate, bilirubin, total oestrogen and growth hormone in the plasma increased, while the concentration of insulin decreased with raising number of fetuses. In comparison with the healthy twin pregnant ewes the animals with subclinical and clinical pregnancy ketosis showed increased values of bilirubin, glutamate-dehydrogenase, growth hormone, total oestrogen, cortisol and cortisol/insulin ratios as well as decreased values of glucose, insulin and potassium. Individual differences of the animals within their groups were observed for the examined hormones as well as for glucose, calcium, sodium and potassium (analysis of variance). Beside multiple pregnancy low insulin values and high cortisol/insulin ratios, which are obviously due to individual disposition, influence enhanced ketogenesis and disorder of glucostasis.
Assuntos
Cetose/veterinária , Complicações na Gravidez/veterinária , Doenças dos Ovinos/etiologia , Animais , Estrogênios/sangue , Feminino , Hormônio do Crescimento/sangue , Hidrocortisona/sangue , Insulina/sangue , Cetose/etiologia , Gravidez , Complicações na Gravidez/etiologia , OvinosRESUMO
Experimentally induced salinomycin toxicosis in weanling pigs showed typical clinical signs of an intoxication with a polyether antibiotic. Severe ataxia and recumbency were the most prominent symptoms, which could be attributed to acute skeletal muscle necrosis by estimation of muscle enzyme activities (creatine kinase, aspartate aminotransferase) and histopathological examination. Intoxication had neither influence on concentrations of vitamin E and selenium-dependent glutathione peroxidase in plasma and different organs nor on contents of fatty acids in skeletal muscles. No signs of increased lipid peroxidation in muscle tissue could be found. Prophylactic application of vitamin E or selenium one day before administration of salinomycin as well as treatment on the following days produced no protective effects. The treated pigs showed equal clinical and pathomorphological alterations as the untreated animals, although applications caused a significant increase of alpha-tocopherol and glutathione peroxidase concentrations in blood and different organs.
Assuntos
Antibacterianos/intoxicação , Piranos/intoxicação , Selênio/uso terapêutico , Doenças dos Suínos/induzido quimicamente , Vitamina E/uso terapêutico , Animais , Intoxicação/tratamento farmacológico , Intoxicação/veterinária , Suínos , Doenças dos Suínos/tratamento farmacológicoRESUMO
The main characteristics of the chemical structure of Deoxyribonucleic acid is described. The methods of sequencing of DNA used today are represented, the base-specific cleavage reactions used in the chemical sequencing procedure and the chain-termination reactions used in the enzymatic procedure. From own studies bovine genomic sequences are shown and some important methodical aspects explained. The application of the sequence analysis is shown with the organization of the porcine growth hormone gene and with the sequence dependence of DNA conformation.
Assuntos
DNA/genética , Código Genético , Animais , Sequência de Bases , Hormônio do Crescimento/genética , Dados de Sequência Molecular , SuínosRESUMO
Early weaned piglets were divided into eight groups of 6 animals each. The animals were fed diets differing in fat quality (4% soybean oil, POZ 5 or 176) and in the content of the vitamins A (5,000 or 20,000 I.U./kg) and E (25 or 125 I.U./kg) over a period of 7 weeks. At the beginning, on day 25 and 47 blood samples were taken and analysed for vitamin A and E. In liver, heart, M. longissimus dorsi and M. semitendinosus vitamin A, E and the TBA-reactive substances were analysed. Induced lipid peroxidation was assessed by the ethane and pentane production rate in the skeletal muscle. During the weaning period a decrease in the alpha-tocopherol level was observed. In groups with the lower doses of vitamin E this effect was more pronounced. After 47 days the alpha-tocopherol concentrations in plasma and heart and skeletal muscle fell about 25-30% by offering high doses of vitamin A compared to those groups fed low doses. Oxidized fats also led to lower tocopherol concentrations in muscle tissues. Hydrocarbon production in M. longissimus dorsi and M. semitendinosus was significantly reduced in groups with the high supplement of vitamin E. A tendentially opposite effect was seen in groups supplied with high levels of vitamin A or oxidized fat. Although retinyl esters in plasma are a minor fraction of the vitamin A activity, they present 99% of the vitamin A in the liver. The distribution pattern of the different retinyl esters was independent of the amount of supplementary vitamin A. In the present experiment 20,000 U of Vitamin A reduced plasma and tissue vitamin E levels. This effect led to an increase of lipid peroxidation indicated by the higher production of hydrocarbons. The results raise concerns about further increases of vitamin A supplementation in piglet feed.
Assuntos
Antioxidantes/metabolismo , Gorduras na Dieta , Suínos/metabolismo , Vitamina A/farmacologia , Vitamina A/farmacocinética , Vitamina E/farmacologia , Vitamina E/farmacocinética , Animais , Alimentos Fortificados , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/metabolismo , Músculo Esquelético/metabolismo , Miocárdio/metabolismo , Óleo de Soja , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Distribuição Tecidual , Vitamina A/sangue , Vitamina E/sangueRESUMO
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.
Assuntos
Deficiências Nutricionais/veterinária , Doenças das Cabras/diagnóstico , Selênio/deficiência , Doenças dos Ovinos/diagnóstico , Deficiência de Vitamina E/veterinária , Animais , Deficiências Nutricionais/sangue , Deficiências Nutricionais/diagnóstico , Doenças das Cabras/sangue , Cabras , Selênio/sangue , Ovinos , Doenças dos Ovinos/sangue , Vitamina E/sangue , Deficiência de Vitamina E/sangue , Deficiência de Vitamina E/diagnósticoRESUMO
In this study consequences of vitamin A-supplementation to the vitamin E-status was investigated in the boar. Three groups of boars, each with 9 animals were fed over a period of seven month with 30000 I.E. Vit. A/kg concentrate (group A), 90 mg b-carotene + 1000 I.E. Vit. A/kg (group B) and 1000 I.E. Vit. A/kg (group C). Every boar was given 100 mg Vit. E/kg plus 50 ml soybean oil/kg to induce oxidative stress. After four month group C showed a higher amount of tocopherol in serum (p < 0.05). The amount of tocopherol in serum of the group B were exactly between group A and C. The amount of retinol in serum of the group C began to decrease after three month due to the high reserve capacity of the liver (p < 0.01). The retinyl ester in serum reflected the state of supply. 90 mg b-carotene led to an efficiency of 15000 I.E. Vit. A. The vitamin antagonism between Vit. A and Vit. E is not based on an antagonism of the intestinal resorption. There was no influence on the daily sperm production caused by different supplementations. The sperm quality was lowered in group C; the number of defective sperm increased (p < 0.001). The supplementation of soybean oil lead to an increase of the saturated fatty acids in the fatty acid pattern of the sperm cells. The increase of saturated fatty acids was the lowest in group C that showed the highest amount of tocopherol in serum.