Cloning and expression of glucose regulated protein 78 (GRP78) in Fenneropenaeus chinensis

GRP78 (78 kDa glucose-regulated protein), also known as BiP (immunoglobulin heavy-chain-binding protein), is an essential regulator of endoplasmic reticulum (ER) homeostasis because of its multiple functions in protein folding, ER calcium binding, and controlling of the activation of transmembrane ER stress sensors. In this report, we cloned the full length cDNA of GRP78 (FcGRP78) from Chinese shrimp Fenneropenaeus chinensis. This cDNA revealed a 2,325 bp with 1,968 bp open reading frame encoding 655 amino acids. This is the first reported GRP78 gene in Crustacea. The deduced amino acid sequence of FcGRP78 shared high identity with previously reported insect GRP78s: 86, 87 and 85% identity with GRP78s of Drosophila melanogaster, Aedes aegypti and Bombyx mori, respectively. Northern blot analysis shows that FcGRP78 is ubiquitously expressed in tissues of shrimp. Heat shock at 35A degrees C significantly enhanced the expression of FcGRP78 at the first hour, reached the maximum at 4 h post heat shock, dropped after that and resumed to the normal level until 48 h of post recovery at 25A degrees C. Additionally, differential expression of FcGRP78 was detected in haemocytes, hepatopancreas and lymphoid organ when shrimp were challenged by white spot syndrome virus (WSSV). We inferred that FcGRP78 may play important roles in chaperoning, protein folding and immune function of shrimp.

Difference in Enzyme Activity and Conformation of Glucose Oxidase Before and After Purification

EEnzyme activity of commercial glucose oxidase was enhanced after purification through a strong anionic exchange resin. In order to get a better insight into this phenomenon, surface pressure–area ( –A) isotherms and surface pressure–time ( –t) isotherms was used to study the interaction and the absorption at different pH values of the subphases between octadecylamine and glucose oxidase purified by a styrene system quaternary ammonium type strongly basic anionic exchange resin. Circular dichroism (CD), electrophoresis and enzyme activity measurements were conducted to study these phenomena. A preliminary hypothesis has been suggested to explain why the enzyme activity of purified glucose oxidase was higher than that of the commercial one. © 2002 Elsevier Science B.V. All rights reserved.

Frog skins keep redox homeostasis by antioxidant peptides with rapid radical scavenging ability

The question of how amphibians can protect themselves from reactive oxygen species when exposed to the sun in an oxygen-rich atmosphere is important and interesting, not only from an evolutionary viewpoint, but also as a primer for researchers interested in mammalian skin biology, in which such peptide systems for antioxidant defense are not well studied. The identification of an antioxidant peptide named antioxidin-RL from frog (Odorrana livida) skin in this report supports the idea that a peptide antioxidant system may be a widespread antioxidant strategy among amphibian skins. Its ability to eliminate most of the 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radical tested within 2 s, which is much faster than the commercial antioxidant factor butylated hydroxytoluene, suggests that it has a potentially large impact on redox homeostasis in amphibian skins. Cys10 is proven to be responsible for its rapid radical scavenging function and tyrosines take part in the binding of antioxidin-RL to radicals according to our nuclear magnetic resonance assay. (C) 2010 Elsevier Inc. All rights reserved.

Enhancement of GABAA Receptor-Mediated Inhibitory Postsynaptic Currents Induced by "Partial Oxygen-Glucose Deprivation

下载PDF阅读器"氧糖剥夺"模型作为研究脑缺血的离体模型被广泛使用,该模型模拟了局灶性脑缺血的主要病理变化.然而在缺血病灶核心区与正常脑组织之间称为缺血半暗带的区域,脑血流也有程度不一的降低.为了模拟这种病理变化,发展了一种"不完全氧糖剥夺"的离体脑片模型,该模型满足两个条件,灌流液里氧气部分剥夺而葡萄糖含量降低;"氧糖剥夺"可以导致谷氨酸介导的兴奋性毒性,从而引起神经细胞的坏死.而A型γ-氨基丁酸受体(GABAAR)介导的神经元抑制性活动可以对抗谷氨酸引起的兴奋性毒性,因此近年来引起广泛的研究兴趣.而谷氨酸受体和γ-氨基丁酸受体功能在缺血半暗带是否有改变尚不得而知.因此本文采用海马脑片全细胞膜片钳的记录方法,研究"不完全氧糖剥夺"对海马CA1区神经元的A型γ-氨基丁酸受体介导的抑制性突触后膜电流(IPSCs)的影响.研究发现"不完全氧糖剥夺"使GABAAR介导的IPSCs的峰值增加而衰减时程延长.进一步研究发现该电流的峰值增加是由于GABAAR-氯离子通道的电导增加所致,而与氯离子的反转电位变化无关.这些发现提示在脑缺血的缺血半暗带区域GABAAR介导的神经元抑制性活动可能是增强的,这可能是神经元面对缺血状态产生自我保护的一种内稳态机制.

Nutritional homeostasis in carnivorous southern catfish (Silurus meridionalis): is there a mechanism for increased energy expenditure during carbohydrate overfeeding?

In previous growth experiments with carnivorous southern catfish (Silurus meridionalis), the non-fecal energy lose was positively related to dietary. carbohydrate level. To test whether metabolic energy expenditure accounts for such energy loss, an experiment was performed with southern catfish juveniles (33.2-71.9 g) to study the effect of dietary carbohydrate level on fasting metabolic rate and specific dynamic action (SDA) at 27.5 degreesC. The fasting metabolic rate in this catfish was increased with dietary carbohydrate level, and the specific dynamic action (SDA) coefficient (energy expended on SDA as percent of assimilated energy) was not affected by dietary carbohydrate level. The results suggest that in southern catfish, carbohydrate overfeeding increases metabolic rate to oxidize unwanted assimilated carbohydrate. A discussion on the poor capacity of intermediate metabolism for adapting dietary carbohydrate in carnivorous fish and its possible relationship with facultative component of SDA was also documented in this paper. (C) 2004 Elsevier Inc. All rights reserved.

Potentiometric glucose sensors

The determination of glucose is possible with the enzymatic reaction of glucose oxidase and potentiometric detection. The signal is proportional to the concentration up to 50 mg/dl. This value is fixed by the concentration of oxygen in the sample. By adding catalase, concentrations up to 2000 mg/dl are detectable. The steepness of the calibration curve is not affected by oxygen concentrations greater than 4 mg/l. In contrast to amperometric sensors, an influence of deposits on the electrodes surface on the signal cannot be found with potentiometric sensors

Effects of temperature, glucose and inorganic nitrogen inputs on carbon mineralization in a Tibetan alpine meadow soil

High levels of available nitrogen (N) and carbon (C) have the potential to increase soil N and C mineralization We hypothesized that with an external labile C or N supply alpine meadow soil will have a significantly higher C mineralization potential and that temperature sensitivity of C mineralization will increase To test the hypotheses an incubation experiment was conducted with two doses of N or C supply at temperature of 5 15 and 25 C Results showed external N supply had no significant effect on CO2 emission However external C supply increased CO2 emission Temperature coefficient (Q(10)) ranged from 113 to 1 29 Significantly higher values were measured with C than with N addition and control treatment Temperature dependence of C mineralization was well-represented by exponential functions Under the control CO2 efflux rate was 425 g CO2-Cm-2 year(-1) comparable to the in situ measurement of 422 g CO2-Cm-2 year(-1) We demonstrated if N is disregarded microbial decomposition is primarily limited by lack of labile C It is predicted that labile C supply would further increase CO2 efflux from the alpine meadow soil (C) 2010 Elsevier Masson SAS All rights reserved

A sensitive NADH and glucose biosensor tuned by visible light based on thionine bridged carbon nanotubes and gold nanoparticles multilayer

A NADH and glucose biosensor based on thionine cross-linked multiwalled carbon nanotubes (MWNTs) and Au nanoparticles (Au NPs) multilayer functionalized indium-doped tin oxide (ITO) electrode were presented in this paper. The effect of light irradiation on the enhancement of bioelectrocatalytic processes of the biocatalytic systems by the photovoltaic effect was investigated.