The decapeptide CMS001 enhances swimming endurance in mice

Now peptides achieve distinct advantages over protein in biological application because of its quick and easy absorption, low power, and high activity. Some bioactive peptides had been developed to be used in the management of exercise-related disorders. In this study, we investigated whether the decapeptide CMS001 (Pro-Thr-Thr-Lys-Thr-Tyr-Phe-Pro-His-Phe) isolated from pig spleen had anti-fatigue effects. Male Balb/c mice were administered CMS001 (20 microg/(kgd)(-1) or 5 microg/(kgd)(-1) for 30 d, intraperitoneal injections) and tested in an exhaustive swim time task. In order to examine the mechanisms of CMS001 anti-fatigue effects, we analyzed liver glycogen stores, blood urea nitrogen (BUN) levels, lactic acid levels, ultrastructural integrity, and levels of both a free radical metabolite and an anti-oxidant enzyme. CMS001 treatment prolonged exhaustive swim time, increased liver glycogen levels, reduced BUN levels, and decreased accumulation of lactic acid in the blood, relative to mice injected with only saline. Examination of the ultrastructure of mitochondria and sarcoplasmic reticulum in skeletal and cardiac muscle of CMS001-treated and control mice revealed that CMS001 can reduce the damage to cardiac and skeletal muscle caused by an exhaustive swim challenge, such that the structure of most tissue specimens were normal in the peptide-treated group. Furthermore the free radical analysis after acute exercise indicated that CMS001 treatment decreased malondialdehyde (MDA) and increased superoxide dismutase (SOD) levels. The present findings indicate that the spleen-derived peptide CMS001 has anti-fatigue effects in mice, and further suggest that the mechanism may involve reduction of tissue damaging free radicals in muscle tissues.     

Cloning and partial characterization of a gene in Bombyx mori homologous to a human adiponectin receptor

In this study, we report the cloning and characteristics of an adiponectin-like receptor gene from Bombyx mori (BmAdipoR) with highly conserved deduced amino-acid sequences and similar structure to the human adiponectin receptor (AdipoR). Structural analysis of the translated cDNA suggested it encoded a membrane protein with seven transmembrane domains. BmAdipoR was found to be expressed in multiple tissues and highly expressed in Malpighian tubules, fat body and testis. BmNPV (Bombyx mori nucleopolyhedrovirus) bacmid system combined with confocal microscopy revealed that BmAdipoR was targeted to the cell membrane. We also found that infection with BmNPV did not have an effect on BmAdipoR mRNA quantity in the midgut of susceptible Bombyx mori strain (306) at 48 h, but BmAdipoR mRNA quantity increased significantly at 72 h. We concluded that BmAdipoR gene was a membrane protein ubiquitously expressed in Bombyx mori tissues and that its expression was altered by treating with BmNPV.     

Distinct control of MyD88 adapter-dependent and Akt kinase-regulated responses by the interleukin (IL)-1RI co-receptor, TILRR

Inflammatory responses are controlled through members of the interleukin-1 receptor (IL-1R)/Toll-like receptor superfamily. Our earlier work demonstrates that the IL-1 receptor type 1 (IL-1RI) co-receptor, Toll-like and IL-1 receptor regulator (TILRR), amplifies IL-1 activation of NF-κB and inflammatory genes. Here we show that TILRR similarly promotes IL-1-induced anti-apoptotic signals and reduces caspase-3 activity. Further, the TILRR-induced effects on cell survival and inflammatory responses are controlled through distinct parts of the IL-1RI regulatory Toll IL-1 receptor (TIR) domain. Alanine-scanning mutagenesis identified a functional TILRR mutant (R425A), which blocked increases in cell survival and upstream activation of Akt but had no effect on amplification of MyD88-dependent inflammatory responses. A second mutant (D448A) blocked TILRR potentiation of MyD88-dependent signals and inflammatory activation but had no impact on cell survival. Secondary structure predictions suggested that the mutations induce distinct alterations in the α-helical structure of the TILRR core protein. The results indicate a role for TILRR in selective amplification of NF-κB responses through IL-1RI and suggest that the specificity is determined by changes in receptor conformation and adapter protein recruitment.     

Overexpressions of dihydroxyacetone synthase and dihydroxyacetone kinase in chloroplasts install a novel photosynthetic HCHO-assimilation pathway in transgenic tobacco using modified Gateway entry vectors

Dihydroxyacetone synthase (DAS) and dihydroxyacetone kinase (DAK) are two key enzymes for formaldehyde assimilation in methylotrophic yeasts. In order to using a Gateway LR recombination reaction to construct a plant expression vector that contains the expression cassettes for the das and dak genes and allow the proteins encoded by the two target genes to be localized to the chloroplasts of transgenic plants, the entry vector pEN-L4*-PrbcS-*T-gfp-L3* contained the tomato rbcS 3C promoter (PrbcS) with its transit peptide sequence (*T) and a GFP reporter gene (gfp) was constructed in this study. To verify the applicability of pEN-L4*-PrbcS-*T-gfp-L3*, we generated an entry vector for the dak gene by replacing the gfp gene in this entry vector with the dak gene. We also generated an entry vector for the das gene by replacing the gus gene in another entry vector (pENTR*-PrbcS-*T-gus) with the das gene. Using these entry vectors and pK7m34GW2-8m21GW3, we successfully constructed the pKm-35S-PrbcS-*T-gfp-PROLD-PrbcS-*T-gus and the pKm-35S-PrbcS-*T-dak-PROLD-PrbcS-*T-das expression vectors. Our results showed that high expression of GUS was achieved in leaves, and the expressed GFP, DAS and DAK proteins could be targeted to the chloroplasts after the two expression vectors were used to transform tobacco. The overexpressions of DAS and DAK in the chloroplasts successfully created a novel photosynthetic HCHO-assimilation pathway in transgenic tobacco. By utilizing these expression vectors, we not only successfully expressed two target genes with one transformation but also localized the expressed proteins to chloroplasts via the transit peptide sequence (*T). Therefore, the construction of pEN-L4*-PrbcS-*T-gfp-L3* establishes a technique platform that provides a convenient means for chloroplast genetic engineering.     

不同产地丹参药材HPLC指纹图谱分析及4种菲醌类成分含量的比较

对7个产地丹参(Salvia miltiorrhiza Bge.)药材进行了HPLC分析并确定了其指纹图谱的共有模式谱,在此基础上比较了7个产地丹参药材中4种菲醌类成分(二氢丹参酮Ⅰ、丹参酮Ⅰ、隐丹参酮和丹参酮ⅡA)的相对含量。结果表明:来源于不同产地的丹参药材的HPLC图谱均有21个峰,其中有8个共有峰;各共有峰的相对保留时间基本相同,RSD值均小于0.086%;但相对峰面积差异明显。以4号峰(二氢丹参酮Ⅰ)为参照峰,初步构建了丹参药材的HPLC指纹图谱的共有模式谱,7个产地丹参药材的HPLC指纹图谱与共有模式谱的相似性良好,相似度值为0.916～0.973。不同产地丹参药材中4种菲醌类成分的相对含量有明显差异,其中,河南产药材中4种菲醌类成分含量均最高,而河北产药材中均最低;此外,不同产地丹参药材中4种成分的组成比例也有明显差异。根据实验结果,建议将丹参药材的HPLC指纹图谱的共有模式谱作为丹参药材质量控制和真伪辨别的标准之一。     

Effects of quorum sensing autoinducer degradation gene on virulence and biofilm formation of Pseudomonas aeruginosa

The aiiA gene from Bacillus thuringiensis was cloned into the Pseudomonas/E. coli shuttle vector and transformed into Pseudomonas aeruginosa strain PAO1. Western blotting showed that the AiiA protein was expressed in PAO1. After induction by IPTG for 6 h and 18 h, expression of the aiiA gene in PAO1 completely degraded the quorum sensing autoinducers N-acylhomoserine lactones (AHLs): N-oxododecanoyl-L-homoserine lactone (OdDHL) and N-butyryl-L-homoserine lactone (BHL). The reduced amount of AHLs in PAO1 was also correlated with decreased expression and production of several virulence factors such as elastase and pyocyanin. AiiA expression also influenced bacterial swarming motility. Most importantly, our studies indicated that aiiA played significant roles in P. aeruginosa biofilm formation and dispersion, as observed by the differences of the biofilm formation on liquid and solid surfaces, and biofilm structures under a scanning electron microscope. These authors contributed equally to this work Supported by the National Natural Science Foundation of China (Grant No. 30570020) and Natural Science Foundation of Hubei Province of China (Grant No. 2004ABA120)     

Technical note phytoremediation of triazophos by Canna indica Linn. in a hydroponic system

The phytoremediation of triazophos (O, O-diethyl-O-(1-phenyl-1, 2, 4-triazole-3-base) sulfur phosphate, TAP) by Canna indica Linn. in a hydroponic system was studied. After 21 d of exposure, the removal kinetic constant (K) of TAP was 0.0229-0.0339 d(-1) and the removal percentage of TAP was 41-55% in the plant system and the K and removal percentage of TAP were about 0.002 d(-1) and 1%, respectively, in darkness and disinfected control. However, the K and removal percentage of TAP were 0.006 d(-1) and approximately 11%, respectively, in the treatment with eluate from the media of constructed wetland. The contribution of plant to the remediation of TAP was 74% and C. indica played the most important role in the hydroponic system. Under the stress of TAP and without inorganic phosphorus nutrient, the activity of phosphatase in the plant system increased and phytodegradation was observed. The production and release of phosphatase is seen as the key mechanism for C. indica to degrade TAP. C. indica, which showed the potential of phytoremediation of TAP, and is commonly used in constructed wetland, so the technique of phytoremediation of TAP from contaminated water can be developed with the combination of constructed wetland.