Stimulation of ROS-scavenging systems in squash (Cucurbita pepo L.) plants by compost supplementation under normal and low temperature conditions

The beneficial effect of compost, the final product of aerobic biodegradation of organic matter, on growth, lipid peroxidation [as malondialdehyde (MDA], hydrogen peroxide (H₂O₂) and superoxide anion (O₂^•−), activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR), as well as reduced ascorbate (ASC) and glutathione (GSH) and their oxidized forms was investigated in squash (Cucurbita pepo L. cv. Eskandarany) plants exposed to normal and low temperature (LT) conditions. LT stress of 8 °C significantly reduced the plant growth of untreated plants, but compost alleviated the adverse effect of stress and significantly increased the fresh and dry weights under normal and stress conditions. LT also induced accumulation of H₂O₂ and O₂^•− and resulted in increased lipid peroxidation, pointing out to cellular oxidative stress. Under compost application, such reactive oxygen species (ROS) and peroxidized lipids were markedly reduced, but SOD, CAT, APX and GR activities, key enzymes of ROS-scavenging systems, were significantly increased. Data also indicated that there were general reductions in total ascorbate and glutathione pool in LT control plants, but compost-treated ones considerably have maintained higher levels of such redox metabolites. Significantly higher ratios of ASC/DHA (dehydroascorbate) and GSH/GSSG (glutathione disulfide) were generally found in compost-treated plants than in untreated-ones. It is evident that compost induced enhancement of LT tolerance was related to up-regulation of enzymatic and non-enzymatic antioxidant systems. Such enhancement would eventually protect plant cells from LT-induced oxidative stress reactions via scavenging ROS.     

Isolation and Characterization of Collagen and Antioxidant Collagen Peptides from Scales of Croceine Croaker (Pseudosciaena crocea)

Acid soluble collagen (ASC) from scales of croceine croaker (ASC-C) was successfully isolated with the yield of 0.37% ± 0.08% (dry weight basis), and characterized as type I collagen on the basis of amino acid analysis and electrophoretic pattern. The antioxidant hydrolysate of ASC-C (ACH) was prepared through a two-stage in vitro digestion (4-h trypsin followed by 4-h pepsin), and three antioxidant peptides (ACH-P1, ACH-P2, and ACH-P3) were further isolated from ACH using ultrafiltration, gel chromatography, and RP-HPLC, and their amino acid sequences were identified as GFRGTIGLVG (ACH-P1), GPAGPAG (ACH-P2), and GFPSG (ACH-P3). ACH-P1, ACH-P2, and ACH-P3 showed good scavenging activities on hydroxyl radical (IC₅₀ 0.293, 0.240, and 0.107 mg/mL, respectively), DPPH radical (IC₅₀ 1.271, 0.675, and 0.283 mg/mL, respectively), superoxide radical (IC₅₀ 0.463, 0.099, and 0.151 mg/mL, respectively), and ABTS radical (IC₅₀ 0.421, 0.309, and 0.210 mg/mL, respectively). ACH-P3 was also effectively against lipid peroxidation in the model system. The antioxidant activities of three collagen peptides were due to the presence of hydrophobic amino acid residues within the peptide sequences. The collagen peptides might be used as antioxidant for the therapy of diseases associated with oxidative stress, or reducing oxidative changes during storage.     

猪ASC及Ub基因的克隆及真核表达重组质粒的构建

炎症小体不仅参与机体的天然免疫反应,同时还参与机体抗病毒等多种宿主防御反应。在多种疾病相关信号通路反应中都伴随着蛋白质的泛素化修饰。NOD样受体(NLRs)介导的炎症小体信号通路中,凋亡相关点状样蛋白(ASC)是关键的接头蛋白。为了解猪瘟病毒(CSFV)感染对ASC泛素化水平的影响,通过RT-PCR从猪血管内皮细胞(ECs)中扩增获得ASC和泛素(Ub)的全长cDNA双链片段,连接至克隆载体pMD-18T,获得重组克隆质粒pMD-ASC和pMD-Ub。双酶切pMD-ASC获得ASC目的序列,连接至pcDNA3.1-Flag,双酶切pMD-Ub获得Ub目的序列,连接至pcDNA3.0-HA,获得重组表达质粒pcDNA3.1-Flag-ASC和pcDNA3.0-HA-Ub。将pcDNA3.1-Flag-ASC和pcDNA3.0-HA-Ub转染ECs,Western blot在蛋白水平检测ASC和Ub的表达。结果表明,成功构建了带有Flag标签的ASC真核表达质粒和带有HA标签的Ub真核表达质粒,且在ECs中获得高效表达,为进一步体外研究CSFV调控猪血管内皮细胞ASC泛素化的机制提供了实验基础。     

Characteristics of Collagen from Rohu (Labeo rohita) Skin

Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) were isolated from rohu skin with the yield of 64.2 and 6.8% (dry weight basis), respectively. Both collagens had glycine as the major amino acid with imino acid content of 196–202 residues/1,000 residues and were characterized as type I collagen with molecular composition of (α1)₂α2-heterotrimer. Fourier transform infrared spectra of both collagens were similar, with no shift in wavenumber of all amide bands. The T_max value of ASC and PSC was 36.40 and 35.48°C, respectively. The zero surface net charge of ASC and PSC was found at pH 5.9 and 5.3, respectively.     

Vitreosity,its stability and relationship to protein content in durum wheat

High quality requirements are set on durum wheat (Triticum durum) from semolina mills and pasta producers. For the production of semolina and pasta with good cooking quality, high grain protein content and vitreosity is required. The dependency of vitreosity on protein content as well as its stability under the influence of humidity was not well investigated up to now. We (1) compared two methods to determine vitreosity, (2) investigated the relationship between vitreosity and protein content, (3) developed a method to analyze vitreosity under humidity, and (4) examined the relationship between protein content and agronomical as well as quality traits in durum wheat. The results showed that the formation of vitreous kernels greatly depends on the protein content. To evaluate the stability of vitreosity under the influence of humidity a new method was elaborated and employed to assess the durum germplasm under study. This revealed that vitreosity of a durum wheat variety depends on the potential to form vitreous kernels but also to maintain this vitreosity under the influence of humidity. Our results further show that protein content is a central trait in durum wheat that strongly influences important traits like grain yield, vitreosity, and b-value.     

Magnesium deficiency induced oxidative stress and antioxidant responses in mulberry plants

The aim of the study was to implicate induction of oxidative stress and antioxidative responses with the effects of Mg deficiency in mulberry plants. Mulberry (Morus alba L.) cv. Kanva-2 plants grown in hydroponics were subjected to deficiency of Mg. Mg-deficient plants developed visible symptoms—deep interveinal chlorotic mottling and necrosis in the older and middle leaves. The decreases in the dry matter yield of plants and concentrations of sugars and starch in the leaves of Mg-deficient plants are suggestive of decreased photosynthetic activity. Mg-deficiency decreased concentrations of photosynthetic pigments, and increased concentrations of H₂O₂ and ascorbate and activities of antioxidative enzymes—peroxidase (POD), ascorbate peroxidase (APX) and superoxide dismutase (SOD). The results suggest induction of oxidative stress by enhancing generation of ROS and inducing alterations in redox status, accompanied by activation of antioxidant machinery including induction of some new SOD isoforms in Mg-deficient mulberry plants. Despite significant increase in H₂O₂, lipid peroxidation was decreased in Mg-deficient plants.     

The Cd‐tolerant rice mutant cadH‐5 is a high Cd accumulator and shows enhanced antioxidant activity

To investigate the mechanism of cadmium (Cd) detoxification in rice (Oryza sativa L.), a Cd‐tolerant mutant cadH‐5, obtained by an Agrobacterium tumefaciens‐based gene‐delivery system, was used for a Cd‐tolerance and accumulation study. After 15 d of exposure to 0.75 mM CdCl₂, significant phenotypic differences were observed between the wild type (WT) and cadH‐5. When exposed to 0.5 mM CdCl₂, higher Cd levels were accumulated in cadH‐5 root cell wall, root cytosol, and membranes than those in WT. However, Cd concentrations in root tissues varied in both WT and cadH5. No significant difference of hydrogen peroxide (H₂O₂) concentrations was observed between WT and cadH‐5, while contents of cell‐wall polysaccharides and phytochelatins (PCs) in the mutant were higher compared to WT. The ratios of reduced glutathione to oxidized glutathione (GSH : GSSG) and ascorbate to dehydroascorbate (ASC : DHA) were lower in WT than in cadH‐5, while the NADPH : NADP⁺ ratio was different to the ratios of GSH : GSSG and ASC : DHA; the ascorbate peroxidase (APX, EC 1.11.1.11), glutathione peroxidase (GR, EC 1.6.4.2), dehydroascorbate reductase (DHAR, EC 1.8.5.1), and monodehydroascorbate reductase (MDHAR, EC 1.6.5.4) activities were lower in WT compared to cadH‐5. Our results indicate that under long‐term Cd stress, cadH‐5 plants can accumulate more Cd with more PC. Also, the redox status of ASC‐GSH cycle was more inhibited in WT than in cadH‐5 plants, rendering WT less able to scavenge reactive oxygen species (ROS). The cadH‐5 mutant maintains relatively high ASC, GSH, and NADPH concentrations, ratios of ASC : DHA, GSH : GSSG, and NADPH : NADP⁺, as well as antioxidative enzymatic activities and PC concentrations. Thus, it is tolerant of relatively high Cd accumulation.     

鲤鱼鳞酸溶性胶原蛋白提取的研究

研究了以酸法提取鲤鱼鳞胶原蛋白过程中温度、时间、乙酸浓度的影响,并以SAS对影响因素进行了回归分析,得到了鲤鱼鳞胶原蛋白提取的最佳条件,温度18℃、提取时间38 h、乙酸浓度为1.3 m ol/L,在提取之前,采用Na2CO3处理对胶原蛋白提取有较大的提高。为淡水鱼综合加工利用提供理论依据。     

L-leucine and L-alanine uptake by trout (Salmo trutta) cardiomyocytes: the effect of IGF-I and insulin

The uptake of L-leucine and L-alanine by trout cardiomyocytes was analysed to identify the carriers involved in their incorporation into cells. The sodium independent systems L and asc, as well as the sodium dependent system ASC, were found to be present. The V_max of these carriers was 33.0, 40.1 and 19.4 pmol amino acid mg⁻¹ prot min⁻¹ for L, asc and ASC, respectively, and the K_m values obtained for these transport systems were 296, 269 and 349 μM amino acid. The L system had as its main substrates the large apolar branched-chain amino acids and showed stereospecificity. The asc and ASC systems carried the short-chain neutral amino acids and were not inhibited by Me-AIB. The ASC transporter also showed adaptive regulation. The effect of IGF-I and insulin on L-alanine uptake, as well as the incorporation of L-leucine into protein, were also studied. Low IGF-I concentrations (below 2.5 ng ml⁻¹) stimulated the uptake of L-alanine and protein synthesis but this effect reverted at high IGF-I concentrations. Insulin depressed protein synthesis and did not affect L-alanine uptake. This revised version was published online in August 2006 with corrections to the Cover Date.     

NLRP3/ASC/caspase-1/IL-1β/IL-18轴在大鼠脓毒症急性肾损伤中的作用

[目的]观察盲肠结扎穿孔(Cecal ligation and puncture,CLP)所致的脓毒症诱导急性肾损伤(AKI)后大鼠不同时间点的血清及肾组织中NLRP3/ASC/caspase-1/IL-1β/IL-18的表达情况。[方法]建立大鼠CLP诱导的脓毒症AKI模型,并于手术后不同时间点用ELISA试剂盒方法测定血清尿素氮(BUN)、血清肌酐(Scr)水平及IL-1β/IL-18的表达量;采用Western blotting法检测肾脏组织NLRP3/ASC/caspase-1/IL-1β/IL-18蛋白水平,并对肾脏组织进行HE染色病理分析。[结果]CLP诱导的脓毒症大鼠血清BUN和Scr水平显著增高,肾小管坏死也显著增加,均于CLP术后24 h达到高峰,与假手术组大鼠相比差异显著(P0.05或P0.01);大鼠肾脏组织中NLRP3/ASC/caspase-1/IL-1β/IL-18及血清IL-1β/IL-18水平也随着损伤程度的加重表现出升高趋势,与假手术组相比差异均达到显著水平。[结论]大鼠脓毒症AKI后肾组织或血清中NLRP3/ASC/caspase-1/IL-1β/IL-18蛋白水平表达增加,表明NLRP3炎症小体的激活可能对脓毒症AKI及其病理改变有促进作用。