Alpha-amylase treatment increases extractable phenolics and antioxidant capacity of oat (Avena nuda L.) flour

In this work, α-amylase was used to treat oat flour with the intent to release phenolic compounds with known antioxidant properties. After methanol extraction, the amounts of nine beneficial phenolic compounds were measured using HPLC. The antioxidant activities of the extracts were assessed using 2,2′-azinobis (3- ethylbenzothiazoline-6-sulfonic acid) (ABTS)，2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP) and protein oxidative damage protection assays. Compared with heating-only treated oat flour, that treated with α-amylase showed significant increase of extractable total phenolic content (0.46–1.35 μmol gallic acid equivalents per gram oat), total antioxidant capacity, and an increased ability on the protection of protein from oxidative damage. Heating-only increased caffeic acid and vanillin content by 17 (0.03 vs 0.54 μg/g oat) and 1.8 (0.62 vs 1.11 μg/g oat) folds, but slightly increased the content of other phenols. Excluding heating effect, α-amylase treatment increased gallic acid content by 2.6 folds (0.38 vs 1.38 μg/g oat), caffeic acid content by 2.4 (0.54 vs 1.82 μg/g oat) folds, and other phenols by 1.0–1.8 folds. In conclusion, α-amylase treatment can yield oat products containing more extractable phenolic compounds with increased antioxidant capacity.     

Immunogenicity of Major Cell Surface Protein(s) of Brucella melitensis Rev 1

Brucella melitensis Rev 1 organisms were salt-extracted and the cell surface proteins (BCSPs) were found to be mainly 39-42 kDa (group 2 porin proteins) in addition to 31.6, 32.5, 58.5 and 14.7 kDa proteins. DEAE-Sephadex anion-exchange column chromatography of BCSPs yielded fraction 1, which contained one major protein (39.8-42.0 kDa) and a minor protein (31.6 kDa). All these proteins were found to be immunogenic by Western blotting. Fraction 1 along with monophosphoryl lipid A and trehalose dicorynomycolate adjuvants as well as BCSPs alone induced significant (p < or = 0.05) protection in BALB/c mice. Both these immunizing agents produced almost equivalent protection to live B. melitensis Rev 1 vaccine at 15 and 30 days post challenge. Lymphocyte stimulation test as well as delayed-type hypersensitivity reaction revealed that both these preparations induced cell-mediated immune response. These preparations also induced humoral immune response as indicated by indirect ELISA. Neither of the immune responses was significantly less (p < or = 0.05) than that with live B. melitensis Rev 1 vaccine, except that their duration was short.     

基于模糊控制的LPG/柴油双燃料电控喷射系统

机械控制的LPG／柴油双燃料发动机难以保证LPG与柴油的最佳掺烧比,满足低碳烟排放,低油耗的要求。本文将模糊控制算法应用于LPG流量的控制,设计了以87C552单片机为核心的模糊智能控制系统,应用于改装后的LPG／柴油双燃料发动机。以发动机转速和油门拉杆位置为输入量,LPG流量为控制量,控制规则设计离线进行,通过控制量决策表对LPG流量进行智能控制,使发动机的碳烟排放大幅降低,燃油消耗有所下降。该系统具有较好的自适应性和鲁棒性。     

20个茶花品种遗传关系的ISSR分析

采用ISSR分子标记技术对茶花品种群中有代表性的20个国内外茶花品种进行了品种间遗传关系的分析.从60对随机扩增引物中筛选出了21对扩增带型清晰及重复性好的引物序列,共扩增出153条带,其中146条呈现多态性,多态性条带比例为95.4%;Nei's基因多样性指数介于0.40～0.48,Shannon信息指数在0.57～0.67,基因分化系数介于0.5～0.7,基因流值介于0.2～0.5,遗传相似系数介于0.50～0.74,平均为0.63.参试的20个茶花栽培品种间遗传差异相对比较窄;结合花型形态学特征与UPGMA聚类分析结果,可将20个茶花品种分为2个大类群.此外,结果显示:ISSR分子标记适用于分析山茶品种间遗传多样性和亲缘关系.     

Effect of TLR2/4 ligand-enhanced non-specific liver inflammation on autoimmune hepatitis in BALB/c mice

AIM: To explore the role of non-specific liver inflammation in inducing autoimmune hepatitis (AIH) in BALB/c mice.METHODS: The plasmids pCYP2D6, pcDNA3.1 and psTLR2/4 were administered by tail vein injection. The carbon tetrachloride (CCl₄) was injected in the abdominal cavity. The autoimmune response was measured by ELISA. The liver inflammation was observed by HE staining. The liver fibrosis was evaluated by Sirius red staining. RESULTS: CCl₄ induced non-specific liver inflammation in the BALB/c mice, and TLR2/4 ligand enhanced the inflammatory responses. After the repeated injection of CCl₄ stopped, the non-specific liver inflammation disappeared, but CCl₄ promoted autoimmune response, autoimmune hepatitis and liver fibrosis induced by mimetic antigen human CYP2D6, and TLR2/4 ligand enhanced these changes. CONCLUSION: TLR2/4-amplified liver non-specific inflammation may play an important role in the initiation and progression of autoimmune hepatitis in BALB/c mice.     

Critical role of cystic fibrosis transmembrane conductance regulator in hypoxia-induced apoptosis of H9c2 cardiomyocytes

AIM:To explore the roles of cystic fibrosis transmembrane conductance regulator (CFTR) in hypoxia-induced apoptosis of H9c2 cardiomyocytes and the underlying mechanisms. METHODS:The rat H9c2 cardiomyocytes were exposed to a 1% hypoxic environment in a hypoxic chamber. After CFTR overexpression, H9c2 cardiomyocytes were cultured in a hypoxic environment. The mRNA and protein levels of CFTR were examined by RT-qPCR and Western blot, respectively. The cell viability was measured by MTT assay. The apoptotic rate was determined by Hoechst 33342 and Annexin V-FITC/PI staining, and the production of reactive oxygen species (ROS) was examined by dichloro-dihydro-fluorescein diacetate (DCF-DA) staining. RESULTS:Hypoxic exposure caused the apoptosis of H9c2 cardiomyocytes, which was accompanied by the down-regulation of CFTR at mRNA and protein levels and over-production of ROS (P<0.05). After CFTR overexpression, the apoptotic rate of the H9c2 cardiomyocytes induced by hypoxia was significantly reduced, with a prominent inhibition of ROS production (P<0.05). However, pretreatment with CFTRinh-172, a specific inhibitor of CFTR, reversed the protective effect of CFTR overexpression in H9c2 cardiomyocytes. CONCLUSION:CFTR has a critical role in protecting against hypoxia-induced apoptosis of H9c2 cells, which may be through inhibiting the generation of ROS.     

Optimisation of Healthy-Lipid Content and Oxidative Stability during Oil Extraction from Squid (Illex argentinus) Viscera by Green Processing

Green extraction was applied to Argentinean shortfin squid (Illex argentinus) viscera, consisting of a wet pressing method including a drying step, mechanic pressing, centrifugation of the resulting slurry, and oil collection. To maximise the oil yield and ω3 fatty acid content and to minimise the oil damage degree, a response surface methodology (RSM) design was developed focused on the drying temperature (45–85 °C) and time (30–90 min). In general, an increase of the drying time and temperature provided an increase in the lipid yield recovery from the viscera. The strongest drying conditions showed a higher recovery than 50% when compared with the traditional chemical method. The docosahexaenoic and eicosapentaenoic acid contents in the extracted oil revealed scarce dependence on drying conditions, showing valuable ranges (149.2–166.5 and 88.7–102.4 g·kg⁻¹ oil, respectively). Furthermore, the values of free fatty acids, peroxides, conjugated dienes, and ω3/ω6 ratio did not show extensive differences by comparing oils obtained from the different drying conditions. Contrary, a polyene index (PI) decrease was detected with increasing drying time and temperature. The RSM analysis indicated that optimised drying time (41.3 min) and temperature (85 °C) conditions would lead to 74.73 g·kg⁻¹ (oil yield), 1.87 (PI), and 6.72 (peroxide value) scores, with a 0.67 desirability value.     

AT1-AA induces autophagy and apoptosis of H9c2 cardiomyocytes through oxidative stress

AIM: To investigate the effects of angiotensin Ⅱ type 1 receptor autoantibody (AT1-AA) on oxidative stress, autophagy and apoptosis in H9c2 cells, and to analyze the possible mechanism. METHODS: The rat H9c2 cells were cultured in vitro. The effect of AT1-AA at different concentrations for different time on the cell viability was measured by CCK-8 assay. Upon the optimum concentration (10^-5 mol/L) and time point (24 h) determined in this stu-dy, the experssion levels of autophagy- and apoptosis-related proteins were detected by Western blot, and the levels of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and malondialdehyde (MDA) were examined by oxidative stress kits. RESULTS: AT1-AA decreased cell viability in a concentration- and time-dependent manner, promoted apoptosis, and up-regulated the levels of autophagy and oxidative stress (P<0.05). The apoptosis of H9c2 cells induced by AT1-AA was decreased after pretreatment with autophagy inhibitor 3-methyladenine (P<0.05). The levels of autophagy and apoptosis in the H9c2 cells pretreated with α-lipoic acid were decreased (P<0.05). Pretreatment with angiotensin Ⅱtype 1 receptor inhibitor telmisartan inhibited oxidative stress, and significantly decreased the levels of autophagy and apoptosis induced by AT1-AA in the H9c2 cells (P<0.05). CONCLUSION: AT1-AA induces autophagy and apoptosis of H9c2 cells through oxidative stress.     

牛巴贝斯虫GST-MSA-2c融合蛋白抗象间接ELISA检测方法的初步建立

以纯化的牛巴贝斯虫GST-MSA-2c融合蛋白作为检测抗原,通过优化ELISA反应条件,初步建立了检测牛巴贝斯虫血清特异性抗体的间接ELISA方法。方阵试验确定的GST-MSA-2C抗原的最适包被浓度为2.5μg／mL,血清最佳稀释倍数为20倍,ELISA阳性反应的临界值为OD450≥0.347,批内和批间重复试验的变异系数均小于10％。经对多例血清检测表明,所建ELSIA检测法重复性好、特异性强、灵敏度高。