Purification and characterization of hepatic triacylglycerol lipase isolated from rainbow trout,Oncorhynchus mykiss

Trialcylglycerol (TG) lipase was isolated and partially purified from rainbow trout liver. Triacylglycerol lipase activity was assayed by measuring¹⁴C-oleic acid release from¹⁴C-triolein.¹⁴C-oleic acid release was linear for up to two hours. Optimal activity occurred at pH 7.0 and 15°C. Most of the lipase activity was recovered in the cytosolic fraction. A 27,000-fold purification was achieved after Sepharose (Bio-gel A 0.5 M, 200–400 mesh) chromatography of a resuspended 20% ammonium sulfate fraction. The molecular weight of the trout hepatic lipase as determined by size-exclusion chromatography and by SDS-polyacrylamide gel electrophoresis was 40–43 kD. Lipase-mediated hydrolysis of TG resulted in the production of diacylglycerols, monoacylglycerols, and fatty acids. Kinetic analysis indicated that V_max=0.016 nmol/h/mg protein and that K_m=0.28 mM triolein. Lipolytic activity was enhanced in the presence of cAMP/ATP-Mg²⁺. These results suggest that the liver of trout possesses a neutral TG lipase that is responsible for mobilizing stored TG and is catalytically activated by phosphorylation.A part of this work was presented at the Annual Meeting of the American Society of Zoologists, December 26–30, 1990, San Antonio, TX.     

Essential Fatty Acids and the Cat

Abstract— The physiology of essential fatty acid metabolism in the cat is reviewed. Emphasis is placed on those aspects of the n:6 and n:3 fatty acids, their metabolites and interactions, which relate primarily to the skin. The functional roles, if known, of the fatty acids are discussed. Recent clinical research into the use of essential fatty acid supplements in the management of feline dermatoses is presented. Current indications for the therapeutic supplementation with essential fatty acids are summarised.     

Diurnal and individual variations in bile acids in the plasma of normal dairy cows

Blood samples were collected every 2 h during a 24 h period from 6 cows of one herd and 10 cows of another herd. In a third herd 9 cows were sampled every 2 h from 6 a.m. to 8 p.m. Concentrations of total bile acids, acetoacetate, glucose and free fatty acids were determined in blood plasma. A marked difference in individual bile acid concentrations and patterns of diurnal variation was found. For most cows the highest bile acid values were observed between 2 and 6 a.m. (overall mean (+/- SD) at 6 a.m.: 104 +/- 84 mumol/l, range: 20-307 mumol/l). Fourteen cows with a bile acid value greater than 90 mumol/l at 6 a.m. ("high BA") were characterized as a group by showing a pronounced decrease in the mean bile acid concentration after morning feeding. In the group of 11 cows with a 6 a.m. bile acid value less than 90 mumol/l ("low BA") the time of day did not contribute significantly to the bile acid variation. For the "high BA" group a nearly synchronous variation between the mean values of the 3 feeding dependent parameters (acetoacetate, glucose and free fatty acids) and the mean values of bile acids was found. The within animal coefficients of correlation between bile acids and the feeding dependent parameters were significantly higher in the "high BA" group than in the "low BA" group. No direct connection was found between bile acid levels and the quantity of concentrates fed or the individual milk yield.     

Effects of β-ME and RA on expression of GFAP in mesenchymal cells derived from mouse fetal liver

AIM: To investigate the effects of β-mercaptoethanol (β-ME) and all-trans rentinal acid (RA) on glial fibrillary acidic protein (GFAP) expression in mesenchymal cells derived from mouse fetal liver in vitro. METHODS: Cells suspension from 14.5-days-old mouse fetal liver were cultured in DMEM/HEPES/F12 supplemented with 20％ FCS and mesenchymal cells were acquired after discarding nonadherent cells. The 5th passage cells were induced by β-ME and RA. The characteristics of treated cells were assayed by immunocytochemistry staining at 5 hours and 5 days after induction. β-actin as an internal control, GFAP gene expression of mesenchyal cells was detected with semi-quantitative RT-PCR. RESULTS: After being inducted by β-ME and RA, 80％ approximately of the cells exhibited typical neural morphology and about 85％ expressed GFAP phenotype. Semi-quantitative RT-PCR showed that mRNA expression of GFAP increased in treated cells versus untreated cells (P<0.01). CONCLUSION: GFAP expression in mesenchymal cells derived from mouse fetal liver in vitro increases after being treated with β-ME and RA.     

Tall oil fatty acid inclusion in the diet improves performance and increases ileal density of lactobacilli in broiler chickens

1. Studies were conducted with tall oil fatty acids (TOFA) to determine their effect on broiler chicken performance and ileal microbiota. TOFA, a product originating from coniferous trees and recovered by fractional distillation of side-streams from pulp production, mainly comprises free long-chain fatty acids (~90%) and resin acids (~8%). Conjugated linolenic acids and pinolenic acid are characteristic fatty acid components of TOFA.

2. TOFA products at 750 mg/kg feed were tested in two 35-day broiler chicken trials, each using a wheat soya-based diet and with 12 replicate pens per treatment. In both trials, TOFA improved body weight gain at all time points (P < 0.001) and feed conversion efficiency during the first 21 days (P < 0.01). Two different dry TOFA formulations (silica carrier and palm oil coating) were tested and showed performance effects similar to liquid TOFA.

3. Ileal digesta of the broiler chickens was analysed for total eubacteria, Lactobacillus spp., Enterococcus spp., Escherichia coli and Clostridium perfringens on days 14 and 35. TOFA significantly increased total eubacteria and lactobacilli density on day 14 (P < 0.05). There was a significant positive correlation between these bacterial groups and broiler body weight on day 14 (P < 0.01).

4. A numerical reduction in C. perfringens was observed. In vitro growth inhibition studies showed that C. perfringens was strongly inhibited by 10 mg/l TOFA (P < 0.001), while common lactobacilli were resistant to >250 mg/l. The in vitro results were thus in line with in vivo observations.

5. The mechanisms behind the bacterial shifts and their role in performance improvement are unknown. Further purification of TOFA components is needed to identify the effective agents.     

美仁牦牛肉不同部位常规营养成分及氨基酸和脂肪酸测定分析

摘 要：选取放牧条件下体况良好、体重相近的甘南成年美仁公牦牛 6 头、母牦牛 4头进行屠宰,采取美仁牦牛肉的肩甲、背最长肌、股二头肌三个部位的分割肉。测定不同部位营养成分、氨基酸和脂肪酸。 结果显示：不同部位美仁牦牛肉的蛋白质含量为22.97%～23.07%,其中股二头肌蛋白质含量最低,背最长肌最高;脂肪含量为1.27%～2.49%,股二头肌脂肪脂肪含量较低,不足1.27%。氨基酸含量丰富,脂肪酸总量依次为肩甲、背最长肌、股二头肌上脑肉。通过对比分析不同部位美仁牦牛肉中营养成分、氨基酸和脂肪酸等含量存在差异。     

BCG诱导RAW264.7细胞脂肪酸氧化对细胞自噬和促炎因子表达的调控作用

旨在探究脂肪酸氧化（fatty acid oxidation,FAO）对BCG介导的RAW264.7细胞自噬和促炎因子表达的调控作用。用BODIPY染色和游离脂肪酸定量试剂盒检测BCG感染后RAW264.7细胞中脂滴聚集情况以及脂肪酸含量;Western blot检测BCG感染对肉毒碱棕榈酰基转移酶1A （CPT-1A）表达的影响;Etomoxir （100 μmol·L^-1）预处理细胞2 h后,BCG感染细胞6 h,检测RAW264.7细胞中BCG存留量,并用Western blot方法检测自噬相关蛋白（Beclin1、LC3-II）和溶酶体蛋白（Rab7）的表达情况;用免疫荧光方法和mRFP-GFP-LC3荧光双标腺病毒分别检测自噬小体聚集和自噬流;荧光定量PCR和ELISA分别检测促炎因子IL-1β、IL-6和TNF-α mRNA表达情况以及在细胞培养上清中的含量。结果显示,BCG感染促进RAW264.7细胞中脂滴聚集和CPT-1A的表达,而游离脂肪酸含量降低;Etomoxir预处理抑制了细胞中BCG存活,并上调了Beclin1、LC3-II和Rab7表达,且细胞中出现大量自噬小体聚集,自噬流增强,却抑制了促炎因子IL-1β、IL-6和TNF-α mRNA表达与分泌。综上表明,抑制FAO可促进BCG感染诱导的RAW264.7细胞自噬,并抑制BCG感染引起的炎症反应。     

Effects of dietary vitamin E and fat supplementation in growing-finishing swine fed to a heavy slaughter weight of 150 kg: II. Tissue fatty acid profile,vitamin E concentrations,and antioxidant capacity of plasma and tissue

The study aimed to assess the effects of vitamin E (VE) supplementation and fat source on fatty acid (FA) composition, VE concentrations, and antioxidant capacity in plasma and tissues of pigs fed to a heavy slaughter weight (150 kg). A total of 64 pigs (32 barrows, 32 gilts; 28.41 ± 0.83 kg) were blocked by sex and weight, and randomly assigned to one of eight dietary treatments (n = 8 per treatment) in a 4 × 2 factorial arrangement. Fat sources included corn starch (CS), 5% tallow (TW), 5% distiller’s corn oil (DCO), and 5% coconut oil (CN); VE supplementation levels were 11 and 200 ppm. Five-phase diets were formulated to meet requirement estimates of NRC (2012) and fed to pigs for each period of 25 kg from 25 to 150 kg. Increasing VE supplementation level increased C16:1 (P < 0.05) content but decreased C20:0 (P < 0.05) content in backfat and belly fat, while in liver, it increased C17:0 (P < 0.05) but decreased C18:0 (P < 0.05). Compared to the pigs fed the CS diet, the pigs fed the CN diet had greater (P < 0.05) content of total saturated FA, the pigs fed the DCO diet had greater (P < 0.05) content of total polyunsaturated FA content and iodine value, and the pigs fed the TW diet had greater (P < 0.05) content of total monounsaturated FA in backfat, belly fat, and liver. Plasma VE concentrations increased linearly (P < 0.05) with increasing length of feeding but faster (P < 0.05) in the pigs fed the CN and TW diets compared with the CS and DCO diets within the 200 ppm VE level; the pigs fed the DCO diet had the highest plasma VE concentrations (P < 0.05) from Phase 2 to Phase 5 within the 11 ppm VE level. The VE concentrations in liver and loin muscle (P < 0.05) increased with increasing dietary VE level from 11 to 200 ppm, but it was not affected by dietary fat source. There was no effect of VE supplementation and fat source on antioxidant capacity in plasma and liver except that pigs fed the DCO diet had greater liver SOD activity (P < 0.05) than the pigs fed the CN diet. In conclusion, dietary VE supplementation did not affect FA profile in backfat, belly fat, and liver consistently, while dietary FA composition with different fat sources affected much of the FA profile in backfat, belly fat, and liver. The higher level of VE supplementation increased liver and muscle VE concentrations and dietary fat sources affected plasma VE concentrations differently (P < 0.05), wherein the TW and CN diets increased the VE absorption greater than the DCO diet.     

低氧适应动物喜马拉雅旱獭的组织学观察

对生活在青藏高原（海拔３２００－３４００m）喜马拉雅旱獭的肝脏、肺、肾、卵巢、甲状腺等器官结构进行了组织学观察。     

High‐level exogenous trans10, cis12 conjugated linoleic acid plays an anti‐lipogenesis role in bovine mammary epithelial cells

Primary bovine mammary epithelial cells (BMECs) were treated by 0, 37.5, 75, 112.5, 150 μmol/L trans10, cis12 conjugated linoleic acid (CLA) to evaluate the effects of different level trans10, cis12 CLA on lipogenesis in BMEC. Addition of 75–150 μmol/L trans10, cis12 CLA reduced significantly the triacylglycerol (TAG) content (P < 0.05), but did not have inhibiting action on cell proliferation (P > 0.05). Treatment with 150 μmol/L trans10, cis12 CLA for 48 h resulted in a 17.1% reduction (P < 0.0001) of medium chain fatty acids (MCFA, C14 < C < C16), a 26.5% reduction (P < 0.0001) of unsaturated fatty acids (UFA) and a corresponding reduction of the mRNA abundance of acetyl coenzyme A (acetylCoA) carboxylase (ACC) (P = 0.046), fatty acid synthase (FAS) (P = 0.017) and stearoylCoA desaturase1 (SCD1) (P = 0.002). Another finding was that trans10, cis12 CLA elevated expression of diacylglycerol acyltransferase2 (DGAT2) (P = 0.020) and long chain acylCoA synthetases (ACSL) (P = 0.032). In conclusion, higher trans10, cis12 CLA, not low trans10, cis12 CLA, inhibited milk fat synthesis and changed fatty acid composition by regulating the expression of FAS, ACC, SCD1, DGAT2 and ACSL.