Development of a novel method to determine very low density lipoprotein kinetics

Isotopic tracer methods of determining triglyceride-rich lipoprotein (TRL) kinetics are costly, time-consuming, and labor-intensive. This study aimed to develop a simpler and cost-effective method of obtaining TRL kinetic data, based on the fact that chylomicrons compete with large VLDL (VLDL(1); S(f) = 60-400) for the same catalytic pathway. Ten healthy subjects [seven men; fasting triglyceride (TG), 44.3-407.6 mg/dl; body mass index, 21-35 kg/m(2)] were given an intravenous infusion of a chylomicron-like TG emulsion (Intralipid; 0.1 g/kg bolus followed by 0.1 g/kg/h infusion) for 75-120 min to prevent the clearance of VLDL(1) by lipoprotein lipase. Multiple blood samples were taken during and after infusion for separation of Intralipid, VLDL(1), and VLDL(2) by ultracentrifugation. VLDL(1)-apolipoprotein B (apoB) and TG production rates were calculated from their linear increases in the VLDL(1) fraction during the infusion. Intralipid-TG clearance rate was determined from its exponential decay after infusion. The production rates of VLDL(1)-apoB and VLDL(1)-TG were (mean +/- SEM) 25.4 +/- 3.9 and 1,076.7 +/- 224.7 mg/h, respectively, and the Intralipid-TG clearance rate was 66.9 +/- 11.7 pools/day. Kinetic data obtained from this method agree with values obtained from stable isotope methods and show the expected relationships with indices of body fatness and insulin resistance (all P < 0.05). The protocol is relatively quick, inexpensive, and transferable to nonspecialist laboratories.     

Analyse,en olfactomètre tabulaire,de l'influence de différents stimulus olfactifs dans la recherche du partenaire sexuel par Zabrotes subfasciatus

Pour rechercher son partenaire sexuel Zabrotes subfasciatus doit utiliser les signaux olfactifs émis soit par les mâles ou par les femelles. Le pouvoir attractif de l'odeur des mâles et des femelles sont donc étudiés et comparés par olfactométrie. Le comportement de mâles et de femelles vierges et inséminées, mis en contact des graines de la plante hôte ou non, est observé en fonction de l'âge des imagos. En aucun cas, les odeurs des mâles n'exercent une attraction sur les femelles. Les femelles vierges en absence de graines de haricot (Phaseolus vulgaris) n'exercent pas d'attraction sur les mâles vierges. Par contre les odeurs des femelles vierges mises en contact avec les graines de haricot sont très attractives pour les mâles vierges et varie en fonction de l'âge: il est plus affirmé chez les femelles plus âgées. L'émission de la phéromone sexuelle est interrompue après l'insémination des femelles mais reprend au bout d'un certain temps après celle-ci.     

黑线仓鼠繁殖输出与基础代谢率的关系

为了解黑线仓鼠繁殖输出与基础代谢率(BMR)的关系,阐明最大持续能量收支(SusMR)的限制水平, 揭示哺乳期能量收支对策,本文测定了哺乳期黑线仓鼠的体重、摄食量、BMR 和身体组成,以及哺乳期的胎仔数、胎仔重和泌乳能量支出(MEO)。结果显示,黑线仓鼠哺乳期体重降低了15.0 ± 0.8% , 摄食量显著增加, 哺乳高峰期平均摄食量为13.9 ± 0.3 g /d, 摄入能为222.1 ± 5.3 kJ/ d, 比哺乳初期增加121% , 比对照组高288% ;哺乳高峰期MEO 为62.4 ± 2.3 kJ/ d, 哺乳末期BMR 为49.7 ± 1.1 kJ/ d; 断乳时平均胎仔数4.7 ± 0.2、窝胎仔重50.5 ±1.6 g; 哺乳末期BMR 比对照组增加48% ,BMR 与消化系统各器官的相关性高于对照组; BMR 与胎仔数、胎仔重、乳腺重量和MEO 显著正相关。结果表明:初次繁殖的黑线仓鼠哺乳期SusMR 限制为4.47 ×BMR, 在自身维持和繁殖输出之间采取了“权衡分配”的原则,通过体重降低以减少BMR 的增加幅度, 从而有利于繁殖输出。     

Bioactive monoterpene indole alkaloids from Nauclea officinalis

Two new monoterpene indole alkaloids, naucleaoffines A (1) and B (2), together with six known alkaloids (3–8), were isolated from the stems and leaves of Nauclea officinalis. The structures of 1 and 2 were elucidated by extensive spectroscopic methods and the known compounds were identified by comparisons with the data reported in literature. All isolated compounds were evaluated for their anti-inflammatory activities and anti-HIV-1 activities. Compounds 1–8 exhibited significant inhibitory activities on nitric oxide (NO) production induced by lipopolysaccharide in mouse macrophage RAW 264.7 cells in vitro with IC₅₀ values comparable to that of hydrocortisone. In addition, compounds 1–8 showed significant anti-HIV-1 activities with EC₅₀ ranged from 0.06 to 2.08 µM. These findings suggest that the discoveries of these indole alkaloids with significant anti-inflammatory activities and anti-HIV-1 activities isolated from N. officinalis could be of great importance to the development of new anti-inflammatory and anti-HIV agents.     

Zooplankton as a compound mineralising and synthesizing system: phosphorus excretion

Data on phosphate excretion rates of zooplankton are based on measurements using the pelagic crustacean zooplankton of Lake Vechten and laboratory-cultured Daphnia galeata. In case of Daphnia sp we measured the effects of feeding on P-rich algae and P-poor algae (Scenedesmus) as food on the P-excretion rates at 20°C. The excretion rates of the natural zooplankton community, irrespective of the influence of the factors mentioned, varied by an order of magnitude: 0.025–0.275µg PO₄-Pmg^–1C in zooplankton (C _zp ) h^–1. The temperature accounted for about half the observed variation in excretion rates. The mean excretion rates in the lake, computed for 20°C, varied between 0.141 and 0.260 µg Pmg^–1C _zp h^–1. Based on data of zooplankton biomass in the lake the P-regeneration rates by zooplankton covered between 22 and 239% of the P-demand of phytoplankton during the different months of the study period.In D. galeata, whereas the C/P ratios of the Scenedesmus used as food differed by a factor 5 in the experiments, the excretion rates differed by factor 3 only. Despite the higher P-excretion rates (0.258± 0.022 µg PO₄-P mg^–1 C h^–1) of the daphnids fed with P-rich food than those fed with P-poor food (0.105 ± 0.047 µg PO₄-P mg^–1 C h^p–1), both the categories of the animals were apparently conserving P. A survey of the literature on zooplankton excretion shows that in Daphnia the excretion rates vary by a factor 30, irrespective of the species and size of animals and method of estimation and temperature used.About two-thirds of this variation can be explained by size and temperature. A major problem of comparability of studies on P-regeneration by zooplankton relates to the existing techniques of P determination, which necessitates concentrating the animals several times above the in situ concentration (crowding) and prolonged experimental duration (starving), both of which manifest in marked changes that probably lead to underestimation of the real rates.     

Influence of age on the bioaccumulation of heavy metals in Apodemus sylvaticus at Merja Zerga lagoon,Morocco

The influence of age and sex on the bioaccumulation of heavy metals in Apodemus sylvaticus was studied in Merja Zerga lagoon in northern Morocco. Five trace metal elements (Zn, Pb, Cr, Cu and Fe) were quantitatively analyzed by Varian AA 240 atomic absorption spectroscopy with graphite furnace in three organs (Liver, Kidney and Heart) from animals of different age and sex. The maximum metal level of the analyzed samples was recorded in adults and was limited to 46.62 μg/g for Pb and 35.1 μg/g for Cu, while it reached 22.69 μg/g, 7.59 μg/g and 6.78 μg/g for Cr, Zn and Fe, respectively. Highly significant differences were found for bioaccumulation of heavy metals according to animal ages and no significant differences were observed between the two sexes among the studied animals. Our results revealed also the existence of a strong correlation (r > 0.65) between the majority of biometric parameters and the trace element concentrations. In general, we found that age is a critical factor in estimating the level of heavy metal pollution. Other characteristics such as habitat, feeding habits and anti-predator behavior of the species need to be studied.     

Up-regulation of GABAB Receptor Signaling by Constitutive Assembly with the K+ Channel Tetramerization Domain-containing Protein 12 (KCTD12)

GABA_B receptors are the G-protein coupled receptors (GPCRs) for GABA, the main inhibitory neurotransmitter in the central nervous system. Native GABA_B receptors comprise principle and auxiliary subunits that regulate receptor properties in distinct ways. The principle subunits GABA_B1a, GABA_B1b, and GABA_B2 form fully functional heteromeric GABA_B(1a,2) and GABA_B(1b,2) receptors. Principal subunits regulate forward trafficking of the receptors from the endoplasmic reticulum to the plasma membrane and control receptor distribution to axons and dendrites. The auxiliary subunits KCTD8, -12, -12b, and -16 are cytosolic proteins that influence agonist potency and G-protein signaling of GABA_B(1a,2) and GABA_B(1b,2) receptors. Here, we used transfected cells to study assembly, surface trafficking, and internalization of GABA_B receptors in the presence of the KCTD12 subunit. Using bimolecular fluorescence complementation and metabolic labeling, we show that GABA_B receptors associate with KCTD12 while they reside in the endoplasmic reticulum. Glycosylation experiments support that association with KCTD12 does not influence maturation of the receptor complex. Immunoprecipitation and bioluminescence resonance energy transfer experiments demonstrate that KCTD12 remains associated with the receptor during receptor activity and receptor internalization from the cell surface. We further show that KCTD12 reduces constitutive receptor internalization and thereby increases the magnitude of receptor signaling at the cell surface. Accordingly, knock-out or knockdown of KCTD12 in cultured hippocampal neurons reduces the magnitude of the GABA_B receptor-mediated K⁺ current response. In summary, our experiments support that the up-regulation of functional GABA_B receptors at the neuronal plasma membrane is an additional physiological role of the auxiliary subunit KCTD12.     

Staining with Fluorescein Diacetate Correlates with Hepatocyte Function

To establish the importance of fluorescein diacetate (FDA) as a viability stain for cultured hepatocytes. we hypothesized that FDA staining would correlate positively with hepatocyte viability and function. Mixtures of live and dead cells were stained with FDA and scanned by flow cytometry. A close correlation was observed between the live cell fraction and percent viability as determined by FDA staining (R² = 0.962). Hepatocytes were also sorted into low fluorescence and high fluorescence groups. Both albumin production and lidocaine metabolism (P-450 activity) were significantly increased in the high fluorescence group compared to the low fluorescence group. An automated, fluorescence-activated assay was useful for rapid assessment of hepatocyte viability. In addition. the intensity of green fluorescence following staining with FDA correlated well with two specific measures of hepatocyte function.     

能量标签技术及其在红树林生态系统能流研究中的应用

传统上认为红树林输出的有机质产生巨大的能流,支持了巨大的河口和近岸水域生态系统的次级生产。但能量标签技术的研究结果却显示红树林输出的有机质的作用并没有如此巨大。用红树碎屑难消化特性来解释此现象,此外数学模型模拟分析发现潮汐的稀释作用也可以解释这种现象。但这两者都不能解释,在其他初级生产者稀少时,红树材输出的有机质可以被大量利用的现象。在有红树林的河口和近海岸水域生态系统中,藻类等非红树初级生产者具有比红树植物更高的初级生产力,而且更容易被动物获得和消化。可以认为是藻类等巨大初级生产力的竞争作用导致红树初级生产在消费者组织中很难被发现,如此上面提到的难题就能得到很好的解决。此外能量标签技术检测出的是红树的初级生产在消费者组织中的相对比率,不是绝对数量值,从此角度看,能量标签技术的结果与传统观点不是矛盾而是互相补充的关系。由此推测红树的初级生产应该还是被消费者所利用,只是它们在消费者初级营养来源组成中占的比例并不大,但其绝对数量并不少。这与传统观点认为的红树的初级生产被大量利用,支撑了具有巨大的次级生产稍有不同。此外,能量标签技术在红树林生态系统中的适用性尚未检验;计算食物组成的数学工具不是很完善;实验设计上考虑的不够全面;对定量研究有一定的影响。     

印度胶树茎、叶制片新尝试

对于印度胶榕这种特殊类的材料制片,在取材、抽气和染色等过程中,常出现抽气不净和染色不均匀及乳汁脱不掉等问题,影响观察效果。从操作程序上进行了改进,取得了明显的效果,对于特殊植物材料制片有一定的参考价值。