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实时荧光核酸恒温检测技术(SAT)在甲型流感病毒动物模型检测中的应用 总被引:1,自引:0,他引:1
评价实时荧光核酸恒温检测技术(SAT)对H1N1和H3N2两种甲型流感病毒小鼠模型肺组织匀浆液的检测效能。小鼠滴鼻感染不同浓度H1N1、H3N2病毒液30μL,感染96h后取肺组织匀浆液分别进行SAT检测及抗原血凝效价测定。结果表明,SAT检测的病毒拷贝数基本与抗原血凝效价测定结果一致,但灵敏度提高10^4倍。与血凝实验比较,SAT检测具有灵敏度高、特异性强、结果准确、高通量自动化的优势,有利于甲型流感动物模型的建立,为实验室筛选抗甲型流感病毒药物奠定基础。 相似文献
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本研究比较不同来源、不同酶解物浓度的家蝇幼虫蛋白及其抗氧化活性。以来自猪粪源和鸡粪源家蝇幼虫为原料制备蛋白粉,用胰蛋白酶水解蛋白粉制备水解产物,以·OH和O_2~-·清除率为指标评价两种蛋白酶解物的抗氧化活性。结果显示,家蝇幼虫蛋白水解物在4 h达水解度最高值,其水解物的·OH清除率在5 h达到最大为59.02%,其水解物的O_2~-·清除率在4 h达到最大为73.24%;随着两种水解物浓度增加,其抗氧化效果越强,当猪粪源和鸡粪源家蝇幼虫酶解物浓度为10 mg/mL时,两者O_2~-·清除率达到最大分别为70%和80%;·OH清除率分别为45%和52%。由此可见,家蝇幼虫酶水解物对超氧阴离子、羟自由基均有清除能力,鸡粪源家蝇幼虫酶解物优于猪粪源。 相似文献
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家蝇作为饲料添加剂对清远良种鸡肉质和风味的影响 总被引:5,自引:0,他引:5
测定蝇蛆和蝇蛹作为饲料添加剂对清远良种鸡肉质和风味的影响,为家蝇Musca domestica L.作为添加饲料改善鸡的肉质和风味,以及代替鱼粉成为新的蛋白质来源提供数据。150只清远鸡雏鸡随机等分成5组,以鱼粉或商业饲料加入不同比例的家蝇幼虫或蝇蛹作为添加饲料加以饲养。结果显示,饲喂不同食料的清远鸡的增重、脾重和法氏囊重影响差异不显著,但饲喂蝇蛆和蝇蛹的鸡群的死亡率明显低于仅饲喂鱼粉或商业饲料的鸡群;饲喂蝇蛆或蝇蛹的鸡肉的系水力(用失水率表示)和风味评价明显提高,氨基酸含量差异显著。说明家蝇幼虫作为动物蛋白可提高鸡的抗疾病能力及改善鸡肉风味。 相似文献
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构建了同时带有乙型肝炎病毒表面抗原基因和大肠杆菌的β-半乳糖苷酶基因的杆状病毒转移载体质粒pVL941lacHBS。应用磷酸钙沉淀技术将pVL941lacHBS DNA转入事先用AcNPV感染过的Sf9细胞,在显色剂X-gal存在下,筛选无多角体的蓝色蚀斑。经过若干次蚀斑纯化,最后获得含乙肝病毒表面抗原基因和β-半乳糖苷酶基因的重组杆状病毒R-AcV941LS。 用R-AcV941LS感染Sf9细胞,在感染后72~96小时,用RPHA和RIA分别检测组织培养上清液和细胞裂解液中的乙型肝炎病毒表面抗原含量。结果,组织培养上清液为3.83μg/1×10~6~2×10~6细胞;细胞裂解液为4.39μg/1×10~6~2×10~6细胞。乙型肝炎病毒表面抗原的合成总量为8.22μg/1×10~6~2×10~6细胞。免疫电镜观察显示表达产物呈约22nm的球形颗粒。小鼠免疫接种实验结果表明,以R-AcV941LS感染Sf9细胞表达的乙型肝炎病毒表面抗原与在哺乳动物细胞系表达的乙型肝炎病毒表面抗原具有相近似的免疫原性。 相似文献
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艾蒿精油对家蝇的忌避及熏蒸活性 总被引:7,自引:0,他引:7
应用“Y”型嗅觉仪对家蝇进行了生物测定。结果表明,艾蒿(Artemisia vulgars L.)精油20μL和25μL剂量对家蝇Musca domestica rieina有很强的忌避活性,平均忌避活力分别为86%和88%,显著高于5和10μL剂量。植物精油对家蝇成虫具有很好的熏蒸活性。随着精油浓度的升高和熏蒸时间的延长,试虫的死亡率显著增加。艾蒿精油浓度为20μL/mL时,家蝇成虫的死亡率在24h后即达到100%;对照在24,48,72和96h时家蝇成虫的死亡率分别为0,0,5%和11%。 相似文献
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《环境昆虫学报》2014,(4):543-548
在实验室条件下研究了三叶蔓荆Vitex trifolia L叶片提取物对甘蓝蚜Brevicoryne brassicae L的拒食、对菜粉蝶Pieris rapae L幼虫的生长发育抑制以及对家蝇Musca domestica的毒杀活性。试验结果表明:10 mg/mL的乙酸乙酯提取物在24 h和48 h对蚜虫的拒食率分别为8889%和8636%,拒食效果明显。几种溶剂提取物对5龄菜粉蝶幼虫的生长发育均有不同程度的影响,其中乙酸乙酯提取物的体重增长抑制率第1天达7535%。各溶剂提取物在高浓度下对家蝇2龄幼虫均有一定的毒杀作用,低浓度下效果较差。乙酸乙酯提取物48 h的LC50为2278 mg/mL,远高于拒食和生长发育抑制试验设定的浓度。结果表明乙酸乙酯提取物拒食活性和生长发育抑制活性显著。 相似文献
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反义寡核苷酸体外抗流感病毒活性 总被引:2,自引:0,他引:2
为了获得具有抗流感病毒活性的反义寡核苷酸,针对A型流感病毒基因组3′和5′端保守序列,设计并合成了多条硫代寡核苷酸(ODN):3′端反义ODN(IV3#)与3′端正义ODN(IV3S);5′端反义ODN(IV4#)与5′端正义ODN(IV4S)以及由5′和3′端正义/反义保守序列组成的复合序列ODN(IV6#和IV7#)。测定了PSODN的体外细胞毒性和在MDCK细胞中对流感病毒复制的影响。结果表明:(1)PSODN浓度高达50μmol/L时对MDCK细胞末表现有毒性作用;(2)与流感病毒基因组5′端互补的ODN IV4#以及由5′和3′端保守序列构成的IV6#ODN和IV7#ODN均具有较高的抗病毒活性;如IV4#ODN浓度为1μmol/L时对流感病毒A/京防/861(H1N1)抑制率近50%,浓度为10μmol/L或更高时抑制率超过70%,且IV4#抑制病毒活性呈现明显的序列和剂量依赖性;(3)IV4#ODN不仅对A型流感病毒H1N1亚型有抑制作用,对H3N2亚型也表现较高的抑制活性;(4)病毒感染复数(MOI)对IV4#ODN抗病毒活性有一定影响,当MOI较低时,IV4#ODN表现的剂量效应关系更加明显。抗流感病毒反义寡核苷核IV4#ODN的发现为进一步研究流感新型药物奠定了实验基础。〖HTH〗关键词〖HTSS〗:流感病毒, 反义寡核苷酸, 体外细胞毒性, 抗病毒活性, 感染复数 相似文献
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Selenium-independent glutathione peroxidase activity associated with glutathione S-transferase from the housefly, Musca domestica 总被引:2,自引:0,他引:2
T W Simmons I S Jamall R A Lockshin 《Comparative biochemistry and physiology. B, Comparative biochemistry》1989,94(2):323-327
1. A glutathione S-transferase having Se-independent glutathione peroxidase activity was isolated from 100,000 g supernatant from housefly homogenate. 2. The specific activity of the partially purified Se-independent glutathione peroxidase was 1776 nmol NADPH oxidized/min/mg protein, representing an 87-fold purification. 3. The Mr of this enzyme was estimated to be 37,000 and 26,000 by gel filtration chromatography and gel electrophoresis, respectively. 4. Selenium-dependent glutathione peroxidase activity could not be detected in this same supernatant. 5. Se-independent glutathione peroxidase activity should be considered in future studies of the insect antioxidant defense system. 相似文献
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溴氰菊酯对不同品系家蝇脑突触体膜蛋白磷酸化及ATP酶活性的影响 总被引:6,自引:2,他引:6
以敏感品系家蝇和溴氰菊酯抗性品系家蝇(Musca domestica L.)为材料,研究和比较了神经毒剂溴氰菊酯对其脑突触体蛋白磷酸化作用的影响。结果表明,浓度为10-5 mol/L溴氰菊酯抑制了敏感品系家蝇脑突触体蛋白磷酸化作用,而对抗性品系家蝇脑突触体蛋白磷酸化作用无明显影响。若反应体系中加入2.5×10-6 mol/L的cAMP显著激活了敏感品系家蝇脑突触体蛋白磷酸化水平,但是当0.6 mmol/Lca2+或0.6 mmol/L Ca2+加10-5 mol/L钙调蛋白时明显增强了抗性品系家蝇脑突触体蛋白磷酸化水平,甚至超过了其对敏感品系的作用水平。此外,还发现不同浓度的溴氰菊酯可抑制突触膜上的Na/K-ATP酶和Ca-ATP酶活力,浓度越高抑制作用也越大,并且敏感品系家蝇对溴氰菊酯的敏感度要高于抗性品系。 相似文献
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Abstract In this paper, we reported the differences in susceptibility to insecticides between adults and larvae of housefly, Musca domestica (L.), and the mechanisms for the differences. The larvae of housefly were much more tolerant to insecticides than the adults, and the tolerance ratio to cyhalothrin was as high as 205.5 for susceptible strain. Mechanism studies showed that higher GST activity was associated with higher insecticide tolerance in the larvae. The co-toxicity coefficient of the mixture of cyhalothrin and methylene dithiocyanate(4:1) on pyrehid-resistant houseflies was 188. 相似文献
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Evaluation of OX40 ligand as a costimulator of human antiviral memory CD8 T cell responses: comparison with B7.1 and 4-1BBL 总被引:5,自引:0,他引:5
Serghides L Bukczynski J Wen T Wang C Routy JP Boulassel MR Sekaly RP Ostrowski M Bernard NF Watts TH 《Journal of immunology (Baltimore, Md. : 1950)》2005,175(10):6368-6377
CTL are important effectors of antiviral immunity. Designing adjuvants that can induce strong cytotoxic T cell responses in humans would greatly improve the effectiveness of an antiviral vaccination or therapeutic strategy. Recent evidence suggests that, in addition to its well-established role in costimulation of CD4 T cell responses, OX40L (CD134) can directly costimulate mouse CD8 T cells. In this study, we evaluated the role of OX40L in costimulation of human antiviral CD8 T cell responses and compared it with two other important costimulators, B7.1 (CD80) and 4-1BBL (CD137L). Delivery of OX40L to human monocytes using a recombinant replication-defective adenovirus led to greater expansion, up-regulation of perforin, enhanced cytolytic activity, and increased numbers of IFN-gamma- and TNF-alpha-producing antiviral memory CD8 T cells in cultures of total T cells. Synergistic or additive effects were observed when OX40L costimulation was combined with 4-1BBL (CD137L) or B7.1 (CD80) costimulation. In total T cell cultures, at low Ag dose, 4-1BBL provided the most potent costimulus for influenza-specific CD8 T cell expansion, followed by B7.1 (CD80) and then OX40L. For isolated CD8 T cells, 4-1BBL was also the most consistent costimulator, followed by B7.1. In contrast, OX40L showed efficacy in direct activation of memory CD8 T cells in only one of seven donors. Thus, OX40L costimulates human antiviral memory CD8 T cell responses largely through indirect effects and can enhance anti-influenza, anti-EBV, and anti-HIV responses, particularly in combination with 4-1BBL or B7. 相似文献
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Ali Nejidat 《Physiologia plantarum》1995,94(3):411-414
Calcium ions play an important role in the regulation of stomatal movement and the mechanism underlying this action is yet to be determined. It is suggested that guard cell plasma membrane ATPase is a target for calcium action and that this effect is mediated by calmodulin. In this study, the effects of calcium and two calmodulin antagonists on ATPase activity in a crude homogenate of Commelina communis L. guard cell protoplasts were examined. The homogenate contained Mg2+ -dependent, K+ -simulated ATPase activity, which was inhibited by CaCl2 while stimulated by the calmodulin antagonists, compound 48/80 and chlorpromazine. The calmodulin antagonists partially reversed the inhibitory effect of calcium ions. The results support the possibility of calmodulin involvement in the regulation of guard cell ATPase activity by calcium ions. 相似文献
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A specially prepared and partially fractionated preparation of gymnemic acid compounds were found to be inhibitory to the ATPase system from housefly brain and labellum, and fish brain. Enzyme activity in a homogenate fraction from labellum showed greatest sensitivity to the gymnemic acid preparation. Na+-K+ ATPase activity from the labellum, which contains the sweet taste sensory receptors, was inhibited by over 92% at a concentration of 4 middot; 10?6M gymnemic acid. Because gymnemic acid compounds are specific suppressants of sweet receptors, the evidence presented in this paper implies that a portion of the ATPase system may be biochemically important in the sweet-sensing mechanism. 相似文献
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为了优化甲醇萃取滞育家蝇Musca domestica幼虫血淋巴中抗病毒肽的方法, 本实验分别比较甲醇及乙腈萃取血淋巴中抗病毒组分效果及最佳使用梯度、上样蛋白浓度、洗脱次数、洗脱流速及操作环境温度, 用Tricine-SDS-PAGE分析甲醇及乙腈的洗脱组分的构成。结果表明:甲醇萃取的活性组分多于乙腈萃取的; 用连续梯度(10%~100%)甲醇萃取的抗病毒活性组分, 比分别用10%, 30%, 50%, 70%, 90%和20%, 40%, 60%, 80%, 100%间断梯度洗脱的回收率高, 同时获得活性组分的重现性好。在0~4℃冰浴的环境下, 于250 mg/6 mL C18萃取柱内加入1.5±0.4 mg/mL蛋白浓度的血淋巴粗提液, 用1 mL/min流速4次洗脱, 抗病毒活性组分可以得到较好的分离效果。 相似文献
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《Insect Biochemistry》1976,6(6):627-629
The transfer ribonucleic acid (tRNA) methylases of the housefly was isolated from different tissues. The activity of this enzyme was tested during the life cycle of the insect. The data show how enzyme activity changes during the course of metamorphosis of the housefly, Musca domestica. 相似文献