Effects of exogenous fatty acids and cholesterol on aminopeptidase activities in rat astroglia

Several studies have addressed the interaction between fatty acids and lipids with central nervous system peptides. Because aminopeptidases (AP) are involved in the regulation of neuropeptides, this work studies several AP expressed in cultured astroglia, after exogenous addition of oleic and linoleic fatty acids and cholesterol to the culture medium. Alanyl-AP, arginyl-AP, cystyl-AP, leucyl-AP, tyrosyl-AP and pyroglutamyl-AP activities were analysed in whole cells using the corresponding aminoacyl-beta-naphthylamides as substrates. Oleic acid inhibits alanyl-AP, cystyl-AP and leucyl-AP activities, whereas linoleic acid inhibits alanyl-AP, arginyl-AP and tyrosyl-AP activities. Neither oleic acid nor linoleic acid modifies pyroglutamyl-AP activity. In contrast, cholesterol increases arginyl-AP, cystyl-AP, leucyl-AP, tyrosyl-AP and pyroglutamyl-AP activities, although it does not modify alanyl-AP activity. The changes reported here suggest that oleic and linoleic fatty acids and cholesterol can modulate peptide activities via their degradation route involving aminopeptidases; each of them being differentially regulated.     

A short-term ingestion of fructo-oligosaccharides increases immunoglobulin A and mucin concentrations in the rat cecum,but the effects are attenuated with the prolonged ingestion

We examined the effects of fructo-oligosaccharides (FOS) on IgA and mucin secretion in the rat cecum after different ingestion periods. Rats were fed a control diet or a diet containing FOS for 1, 2, 4, and 8 wk. FOS ingestion greatly increased IgA and mucin concentrations at 1 and 2 wk, but the effects were disappeared or attenuated at 4 and 8 wk. After 1 wk, FOS induced higher lactobacilli and lactate concentrations and lower cecal pH in the cecum, but the alterations were moderated with the prolonged ingestion accompanying with increasing short-chain fatty acid concentrations. At 1 and 2 wk, FOS increased IgA plasma cells and polymeric immunoglobulin receptor expression in the cecal mucosa and strongly depressed fecal mucinase activities related to the lower cecal pH. These findings may explain the FOS-induced early elevation of IgA and mucin. Clearly, FOS effects on IgA and mucin secretion considerably differ depending on the ingestion period.     

盐度对黄鳝摄食节律和排粪时间的影响

室内研究了不同盐度(0、2、4、6、8、10和12 g·L^-1)对经过摄食驯化的黄鳝摄食节律和排粪时间的影响.结果表明：盐度对黄鳝的摄食节律没有显著影响（P>0.05）,但不同时段的摄食比例存在极显著差异(P<0.01),每昼夜只有1个摄食高峰期.黄鳝排粪分批次完成,不同盐度下其批次不同.在盐度为0（对照）、2、8、10和12 g·L^-1条件下黄鳝均为3批排粪,在盐度4和6 g·L^-1下排粪批次分别为4批和5批.不同盐度下黄鳝的同批排粪时间均存在极显著差异(P<0.01),对照组的前3批排粪时间极显著高于其它组(P<0.01),即对照组前3批排粪相对推迟4～25 h.研究结果将为盐碱、滩涂水域人工养殖黄鳝的摄食管理和水质调控提供理论依据.     

Delineation of molecular changes in intrahepatic cholesterol metabolism resulting from diminished cholesterol absorption

The absorption of cholesterol by the small intestine is a major route for the net entry of cholesterol into the body and can therefore affect the plasma low density lipoprotein-cholesterol (LDL-C) concentration. These studies used ezetimibe, a potent inhibitor of cholesterol absorption, to delineate the biochemical and molecular changes in intrahepatic metabolism and biliary lipid secretion when there is a major reduction in chylomicron cholesterol delivery to the liver. In female LDL receptor (LDLR)-deficient (LDLR-/-) mice fed a basal diet containing ezetimibe (0-10 mg/day/kg body weight), cholesterol absorption was reduced up to 91%, fecal neutral sterol excretion was increased up to 4.7-fold, and plasma total cholesterol concentrations decreased by up to 18%. Blocking cholesterol absorption prevented the accumulation of very low density lipoproteins and LDL in the circulation of LDLR-/- mice fed a lipid-rich diet. In female LDLR+/+ mice fed the lipid-rich diet with ezetimibe, the relative mRNA level for the LDLR in the liver was 2-fold greater than in matching mice given the lipid-rich diet alone. We conclude that in the mouse the reduction in plasma LDL-C levels induced by blocking cholesterol absorption reflects both a diminished rate of LDL-C production and a modest increase in hepatic LDLR expression.     

Effects of bird ingestion on seed germination of Sorbus commixta

To determine the effects of ingestion by birds on seed germination, we performed germination experiments in the field and laboratory with Sorbus commixta. The germination of four groups of seeds was compared: ingested seeds, seeds defecated in feces after feeding of fruits to birds; extracted seeds, seeds deliberately extracted from the fruit pulp; juiced seeds, seeds plus the juice of the pulp after seeds had been deliberately extracted from the pulp; intact seeds, seeds in untreated intact fruits. In the laboratory, intact and juiced seeds hardly germinated, but ingested and extracted seeds germinated. Thus, the pulp and its juice appeared to inhibit germination, but seeds could germinate without ingestion by birds once the seeds had been manually extracted from the pulp. In the field, intact fruits did not germinate in the first spring, because the seed was still covered with pulp. The pulp of intact seeds decomposed during the first summer, and thus, the seeds had the potential to germinate during the second spring. In fact, most intact seeds do not germinate during the second spring either, since they lose their viability during the first summer. Thus, under natural conditions, most seeds of Sorbus commixta cannot germinate without bird ingestion. Received: 5 July 1997 / Accepted: 7 November 1997     

An experimental analysis of ingestion rates in an omnivorous species

Food intake is difficult to estimate under natural conditions. We investigated ingestion rates of 14 different food types in 26 captive capuchin monkeys (Cebus apella). The procedure consisted in weighing a piece of food and using a two alternative choice tests to present food to the subject, alone in its cage. We recorded the food chosen and the time it took the subject to consume the food entirely. Consumption time was converted into ingestion rates (g/s) for each food type. Ingestion rates of food types significantly differed, and the difference was significantly higher among foods than among subjects. In particular, ingestion rates of the fruits were higher than those of human-processed food. Interestingly, food preferences were significantly related to energy intake rate, i.e., to the amount of energy ingested per unit of time, but not with ingestion rates or energy content alone. The energy acquired by eating different types of food cannot be calculated on the basis of the time spent eating unless a correction factor for each given food (or similar ones) is applied. Future controlled studies should provide field researchers with such corrections factors, possibly using foods collected in the wild.     

Anacetrapib promotes reverse cholesterol transport and bulk cholesterol excretion in Syrian golden hamsters

Cholesteryl ester transfer protein (CETP) transfers cholesteryl ester (CE) and triglyceride between HDL and apoB-containing lipoproteins. Anacetrapib (ANA), a reversible inhibitor of CETP, raises HDL cholesterol (HDL-C) and lowers LDL cholesterol in dyslipidemic patients; however, the effects of ANA on cholesterol/lipoprotein metabolism in a dyslipidemic hamster model have not been demonstrated. To test whether ANA (60 mg/kg/day, 2 weeks) promoted reverse cholesterol transport (RCT), 3H-cholesterol-loaded macrophages were injected and (3)H-tracer levels were measured in HDL, liver, and feces. Compared to controls, ANA inhibited CETP (94%) and increased HDL-C (47%). 3H-tracer in HDL increased by 69% in hamsters treated with ANA, suggesting increased cholesterol efflux from macrophages to HDL. 3H-tracer in fecal cholesterol and bile acids increased by 90% and 57%, respectively, indicating increased macrophage-to-feces RCT. Mass spectrometry analysis of HDL from ANA-treated hamsters revealed an increase in free unlabeled cholesterol and CE. Furthermore, bulk cholesterol and cholic acid were increased in feces from ANA-treated hamsters. Using two independent approaches to assess cholesterol metabolism, the current study demonstrates that CETP inhibition with ANA promotes macrophage-to-feces RCT and results in increased fecal cholesterol/bile acid excretion, further supporting its development as a novel lipid therapy for the treatment of dyslipidemia and atherosclerotic vascular disease.     

The effect of food composition on ingestion,development, and survival of a harpacticoid copepod, Tisbe cucumariae Humes

Feeding experiments were carried out on the benthic harpacticoid copepod Tisbe cucumariae Humes, using seven different diets of various dried and ground macroalgae and marsh grass, algal Aufwuchs, diatoms, polychaete meat, and cereal. In short-term experiments (1 h), ¹⁴C-labelled foods were used to measure ingestion rate of non-ovigerous adult females (individual dry wt = 5.57 ± 2.49 μg). No significant difference was found among the rates at which all foods, except polychaete meat, were ingested (12.7 to 17.3 × 10^?2μg dry wt· ind^?1· h^?1). Polychaete meat was consumed faster (23.9 × 10^?2μg dry wt·ind ^?1· h^?1). The nutritional value of the foods was estimated in long-term experiments (22 days) by measuring development time and survival of T. cucumariae. Both these variables were significantly correlated with the nitrogen, protein content, and C:N ratio of the foods. No relation was found, however, with the amount of carbon, calories and available calories in the diets. Thus, nitrogen (protein) content of the food was the factor limiting secondary production of the copepods.     

Hepatic overexpression of cholesteryl ester hydrolase enhances cholesterol elimination and in vivo reverse cholesterol transport

Neutral cholesteryl ester hydrolase (CEH)-mediated hydrolysis of cellular cholesteryl esters (CEs) is required not only to generate free cholesterol (FC) for efflux from macrophages but also to release FC from lipoprotein-delivered CE in the liver for bile acid synthesis or direct secretion into the bile. We hypothesized that hepatic expression of CEH would regulate the hydrolysis of lipoprotein-derived CE and enhance reverse cholesterol transport (RCT). Adenoviral-mediated CEH overexpression led to a significant increase in bile acid output. To assess the role of hepatic CEH in promoting flux of cholesterol from macrophages to feces, cholesterol-loaded and [(3)H]cholesterol-labeled J774 macrophages were injected intraperitoneally into mice and the appearance of [(3)H]cholesterol in gallbladder bile and feces over 48 h was quantified. Mice overexpressing CEH had significantly higher [(3)H]cholesterol radiolabel in bile and feces, and it was associated with bile acids. This CEH-mediated increased movement of [(3)H]cholesterol from macrophages to bile acids and feces was significantly attenuated in SR-BI(-/-) mice. These studies demonstrate that similar to macrophage CEH that rate-limits the first step, hepatic CEH regulates the last step of RCT by promoting the flux of cholesterol entering the liver via SR-BI and increasing hepatic bile acid output.     

Stimulatory effect of regucalcin on proteolytic activity is impaired in the kidney cortex cytosol of rats with saline ingestion

The effect of regucalcin (RC) on neutral proteolytic activity in the cytosol of rat kidney cortex was investigated. Proteolytic activity was significantly increased by the presence of RC (0.01 + 0.10 M) in the enzyme reaction mixture. This increase was completely abolished by the addition of anti-RC monoclonal antibody (150 ng/ml). When the renal cortex cytosol was incubated without RC addition, the degradation of globin of substrate was demonstrated by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. This degradation was clearly inhibited by the addition of anti-RC antibody (150 ng/ml), indicating that protein degradation results partly from the cytosolic endogenous RC. Meanwhile, proteolytic activity was significantly decreased in the renal cortex cytosol of rats with saline ingestion for 2, 7, and 14 days. The effect of RC (0.1 M) in increasing proteolytic activity was weakened in the kidney cortex cytosol of saline-ingested rats. The present study suggests that endogenous RC plays a role in the activation of proteases in the renal cortex cytosol, and that the RC effect is impaired in saline-ingested rats.     

鸭类摄食对海三棱藨草种子萌发的影响

研究了鸭类的消化作用对海三棱藤草（Scirpus mariqueter）萌发速度和萌发率的影响。模拟消化道的环境(酸性条件pH=2,高温T=42℃,和消化道对种子的研磨作用),分别对海三棱蔗草的种子进行了处理,并与自然环境下鸭类未取食的种子及鸭类粪便中采集的种子进行比较。结果表明。研磨作用能够显著加快种子的萌发速度,而酸性环境和高温条件减缓了种子的萌发速度。鸭类的消化作用对海三棱藤草种子的萌发率没有显著影响。     

Regulation of acyl CoA: Cholesterol acyl transferase (ACAT) activity by mevalonate and cholesterol in isolated rat hepatocytes during perinatal development

Acyl CoA: cholesterol acyl transferase (ACAT) activity presents marked oscillations and differential sensitivity to the in vitro stimulation of the kinase-phosphatase modulatory system in the perinatal rat liver.The regulation of this enzyme activity by some modulators generally active in adulthood, such as cholesterol, lipoproteins and mevalonate, has been studied in hepatocytes isolated at different developmental stages. A lack of effect of mevalonate and a positive effort of lipoprotein cholesterol have been observed at the fetal and neonatal stages.A differential prevalence is suggested of one of the two modulatory mechanisms (phosphorylation-dephosphorylation system, or substrate effect) at each developmental stage.     

Endogenous cholesterol synthesis is sufficient for ACTH-induced differentiation of rat adrenocortical cells in primary culture

Summary To define the role of endogenously synthesized cholesterol in the differentiation of adrenocortical cells in primary culture, fetal rat adrenal cells were cultured in the presence of exogenous cholesterol (serum-supplemented medium) or in the absence of it (serum-free medium or lipoprotein-free medium). Ultrastructurally the cells had features of glomerulosa cells: mitochondria were oval or rod shaped with lamellar inner membranes. The amount of smooth endoplasmic reticulum was small, and lipid droplets were few. When the cells were cultured in serum-free medium some intracytoplasmic vacuoles were seen. The undifferentiated zona glomerulosa-like cells secreted low amounts of corticosterone and 18-OH-deoxycorticosterone (18-OH-DOC) in all three media (serum-supplemented medium, serum-free medium, and lipoprotein-free medium). Stimulation of the adrenocortical cells with ACTH induced the ultrastructural features of differentiated zona fasciculata-like cells. Mitochondrial inner membranes were well developed in lipoprotein-free medium, but not in serum-free medium. The amount of intracellular lipids was increased in both media devoid of cholesterol. In the ACTH stimulated cultures the presence of exogenous cholesterol resulted in increased secretions of corticosterone and 18-OH-DOC. In the absence of an exogenous source of cholesterol, the amounts of steroids secreted were only half of that secreted in the presence of serum-supplemented medium. Endogenously synthesized cholesterol is sufficient for the morphologic differentiation of fetal rat adrenocortical cells under ACTH stimulation. However, without exogenously provided cholesterol, the steroid production accounts only for half of the maximal output achieved using serum-supplemented medium. This work was supported by Finnish Culture Foundation.     

His-tag对胆固醇氧化酶基因在大肠杆菌中表达的影响

为考察组氨酸标签(His-tag)对Brevibacterium sp.DGCDC-82中胆固醇氧化酶基因(ChoAb)在大肠杆菌中表达的影响,将PCR扩增后得到的结构基因与pET28a(+)连接,构建重组质粒pETChoAb(不带His-tag),pETChoAbn(His-tag位于N端)和pETChoAbc(His-tag位于C端)并在大肠杆菌中进行表达.对重组酶进行酶活检测,结果表明His-tag位于ChoAb的C端和N端,COD单位体积酶活由未带标签时的1.72 U/mL分别提高到4.03 U/mL和11.36 U/mL.利用软件Quantity One对SDS-PAGE电泳条带进行灰度分析,结果显示与不带His-tag的COD相比,His-tag位于ChoAb的C端和N端,COD表达量由8.8％增加到16.4％与72.3％.同时菌体浓度分别提高了1.2倍和3.2倍.作为纯化标签,该研究结果对His-tag用于诊断用酶COD的分离纯化可以提供一定的理论指导.     

Effects of geography,age, and gender on Chinese children's soil ingestion rate

The soil/dust ingestion rate (SIR) is a crucial parameter for a human health risk assessment associated with contaminated soil. In addition, SIR levels may be influenced by demographic characteristics and geographic environments. The objectives of this study were to explore the effects of geography, age, and gender on Chinese children's SIR levels. We estimated SIRs for 210 children aged 2.5 to <12 years from three cities of China. The results showed that the mean SIR level for children aged 2.5 to <6 years (kindergarten age) was 60.8 mg d^?1, significantly lower than that (91.6 mg d^?1) observed for children aged 6 to <12 years (primary school age). There was no significant difference in mean SIR levels between females (78.0 mg d^?1) and males (69.2 mg d^?1). The mean SIR level (117.4 mg d^?1) for the children group in Lanzhou was significantly higher than that for the Wuhan children group (63.0 mg d^?1) and the Shenzhen children group (47.5 mg d^?1). However, there was an appreciable but not statistically significant difference in SIR levels between children groups in Wuhan and Shenzhen. Our study highlights the necessity of developing SIR values for children in different geographic locations and age stages.     

Age-dependent changes in rat liver lipid peroxidation and glutathione content induced by acute ethanol ingestion

The study of the influence of the age of animals (13 to 53 weeks) on total liver thiobarbituric acid reactive substances (TBAR) content showed an increase which is maximal in rats of 39 weeks of age compared to young animals (13 weeks), followed by a dimunition in the 53 weeks old group. In this situation, the content of hepatic GSH and total GSH equivalents as well as the GSH/GSSG ratio were decreased with ageing, while GSSG levels were enhanced in the oldest group studied. Acute ethanol intoxication resulted in a marked increase in liver TBAR content in young animals, together with a decline in GSH, total GSH equivalents and GSH/GSSG ratio, and an enhancement in GSSG. These changes elicited by ethanol intake were reduced with ageing. It is concluded that ethanol-induced oxidative stress in the liver is diminished during ageing, despite the progressive decrease in the glutathione content of the tissue observed in control animals.     

In vivo tissue cholesterol efflux is reduced in carriers of a mutation in APOA1

Atheroprotection by high density lipoprotein (HDL) is considered to be mediated through reverse cholesterol transport (RCT) from peripheral tissues. We investigated in vivo cholesterol fluxes through the RCT pathway in patients with low plasma high density lipoprotein cholesterol (HDL-c) due to mutations in APOA1. Seven carriers of the L202P mutation in APOA1 (mean HDL-c: 20 ± 19 mg/dl) and seven unaffected controls (mean HDL-c: 54 ± 11 mg/dl, P < 0.0001) received a 20 h infusion of ¹³C₂-cholesterol (¹³C-C). Enrichment of plasma and erythrocyte free cholesterol and plasma cholesterol esters was measured. With a three-compartment SAAM-II model, tissue cholesterol efflux (TCE) was calculated. TCE was reduced by 19% in carriers (4.6 ± 0.8 mg/kg/h versus 5.7 ± 0.7 mg/kg/h in controls, P = 0.02). Fecal ¹³C recovery and sterol excretion 7 days postinfusion did not differ significantly between carriers and controls: 21.3 ± 20% versus 13.3 ± 6.3% (P = 0.33), and 2,015 ± 1,431 mg/day versus 1456 ± 404 mg/day (P = 0.43), respectively. TCE is reduced in carriers of mutations in APOA1, suggesting that HDL contributes to efflux of tissue cholesterol in humans. The residual TCE and unaffected fecal sterol excretion in our severely affected carriers suggest, however, that non-HDL pathways contribute to RCT significantly.