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.     

Reproductive biology of the female Japanese mantis shrimp Oratosquilla oratoria (Stomatopoda) in relation to changes in the seasonal pattern of larval occurrence in Tokyo Bay, Japan

ABSTRACT:   The reproductive traits and the monthly larval abundance of the mantis shrimp Oratosquilla oratoria were investigated in Tokyo Bay, Japan, in 2002. The goal of the study was to elucidate the cause of changes in the monthly pattern of larval abundance from the 1980s to the 1990s as these changes relate to variation in the stock size of the adult shrimp. Oogenesis was divided into 10 stages by histological observation. The developmental stage of oocytes in an individual's ovary was synchronous, suggesting that almost all the oocytes in an ovary are spawned at the same time. The size at first maturity was estimated to be 7 ≤ body length ( BL ) < 8 cm. Fecundity was expressed as a function of BL , ranging from 19 300 eggs for 8 cm BL to 92 100 eggs for 14 cm BL . Small female shrimps (<10 cm BL ) spawned around August. Most large female shrimps (≥10 cm BL ) spawned around May, and some large female shrimps also spawned until September. Although most large female shrimps spawned in spring, the larval abundance was low before July and high from August onwards. The results suggest that a substantial decrease in the stock size of large individuals causes the low larval abundance before July.     

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.     

Sca-1+ cells from mouse fetal liver can differentiate into neurons in vitro

AIM: To study whether Sca-1⁺ cells from fetal liver can be induced to differentiate into neuronal cells in vitro. METHODS:Sca-1⁺cells from 14 5-days-old murine fetal liver were isolated with a magnetic cell sorting kit, and were cultured in Dulbecco s modif ied Eagle s medium(DMEM)/F12 supplemented with 10%fetal bovine serum(FBS), and passaged at a rat io of 1 3 when cells reached more than 80%confluence.The 5 passage cells were induced by 10^-3mol/Lβ-mercaptoethanol(β-ME)and 5×10^-7 mol/L all-trans-retinoic acid(RA)for 24 hours, and then incubated in serum-free medium for 5 hours to 5 days.The characteristics of treated cel s were assayed by immunocytochemistry staining analysis at 5 hours, or 5 days.RESULTS: Cells treated with β-ME and RA exhibited neuronal phenotype and expressed neuron-specific protein such as neuron-specific nuclear protein (NeuN), neuronfilament-M, and neuron-specific tubulin-1 (TuJ-1) but not tau, MAP-2, or the astrocyte-specific marker glial fibrillary acidic protein (GFAP).CONCLUSION: Sca-1⁺ cells from fetal liver, of which most are regarded as hematopoietic stem cells, could differentiate into early immature neuronal cells in vitro. These findings suggest that Sca-1⁺ cells from fetal liver may be an alternative source in cell therapy and gene therapy of neural dysfunction.     

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.     

Effect of Discontinuous Administration of β -glucan and Glycyrrhizin on the Growth and Immunity of Pacific White Shrimp Litopenaeus Vannamei

A six-week growth trial was conducted to compare the effects of different feeding strate- gies of dietary immunostimulants on the growth and immunity of white shrimp Litopenaeus vannamei （4.70 ±0.20g）. Shrimps were fed with diet containing glycyrrhizin continuously, containing β -glucan continuously, discontinuously （seven days with diet containing β -gluseven days with diet without -glucan; two days with diet containing β-glucan following five days with diet without -glucan）,     

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。     

连续低剂量黄曲霉毒素B1对大鼠肝肾损伤作用

用含有不同质量浓度黄曲霉素B1（AFB1）（0、4、20、25、50μg／kg）的污染饲料连续饲喂各试验组大鼠60d,于7、15、30、60d静脉取血,测定各组大鼠肝功能与肾功能生化指标。每周检测大鼠体质量,试验第60天断头处死大鼠,检测肝肾组织病理学变化并计算各组大鼠的脏体系数。试验期间备组受试动物未发现明显的中毒症状和异常情况,60d内无受试动物死亡。与对照相比,不同剂量组AFB1对大鼠体质量无显著性影响;连续喂饲7d,大鼠血清AST和BuN含量显著升高（P〈0．05）;连续喂饲15d,大鼠血清AsT、BUN和ALB含量显著升高（P〈0．05）,连续喂饲30d,大鼠血清BUN含量显著升高（P〈0．05）,连续喂饲60d,大鼠血清CR和BUN含量显著升高（P〈0．05）。在各测试质量浓度条件下,大鼠虽未发现明显的中毒症状和异常情况,试验期间无动物死亡,但与空白对照相比,各剂量组AFBl对大鼠均产生了显著的肝肾组织病理学损伤,中毒程度与给毒剂量成正相关性。随着毒素暴露时间延长,大鼠肝脏对低剂量AFBl表现出一定的耐受性,而大鼠的肾脏损伤与毒素暴露时间表现出显著的积累效应。     

何首乌对兔血脂及脂肪肝形成的影响

为观察何首乌对高脂饮食兔血清脂质代谢和脂肪肝形成的影响,采用高脂饮食建立兔高脂血症模型,动态检测对照组、高脂模型组、何首乌高剂量组、中剂量组和低剂量组血清中TC、TG、LDL—C、HDL—C含量,实验8周末进行肝脏病理组织学观察。结果：何首乌高、中、低剂量组能明显降低高脂饮食兔血清中TC、TG、LDL—C含量（P〈0．01）,并能提高HDL—C含量（P〈0．05）;光镜下何首乌高、中剂量组肝脏脂肪病变减轻,肝细胞脂变率明显下降（P〈0．05）。结果表明,何首乌具有一定的调节血脂,抑制高脂饮食所致兔脂肪肝形成的作用。