45 resultados para CAT EYE SYNDROME
Resumo:
The paper describes the rapid and label-free detection of the white spot syndrome virus (WSSV) using a surface plasmon resonance (SPR) device based on gold films prepared by electroless plating. The plating condition for obtaining films suitable for SPR measurements was optimized. Gold nanoparticles adsorbed on glass slides were characterized by transmission electron microscopy (TEM). Detection of the WSSV was performed through the binding between WSSV in solution and the anti-WSSV single chain variable fragment (scFv antibody) preimmobilized onto the sensor surface. Morphologies of the as-prepared gold films, gold films modified with self-assembled alkanethiol monolayers, and films covered with antibody were examined using an atomic force microscope (AFM). To demonstrate the viability of the method for real sample analysis, WSSV of different concentrations present in a shrimp hemolymph matrix was determined upon optimizing the surface density of the antibody molecules. The SPR device based on the electroless-plated gold films is capable of detecting concentration of WSSV as low as 2.5 ng/mL in 2% shrimp hemolymph, which is one to two orders of magnitude lower than the level measurable by enzyme-linked immunosorbant assays. (c) 2007 Elsevier B.V. All rights reserved.
Resumo:
For goldfish (Carassius auratus), there are many varieties with different eye phenotypes due to artificial selection and adaptive evolution. Dragon eye is a variant eye characterized by a large-size eyeball protruding out of the socket similar to the eye of dragon in Chinese legends. In this study, anatomical structure of the goldfish dragon eye was compared with that of the common eye, and a stretching of the retina was observed in the enlarged dragon eye. Moreover, the homeobox-containing transcription factor Six3 cDNAs were cloned from the two types of goldfish, and the expression patterns were analyzed in both normal eye and dragon eye goldfish. No amino acid sequence differences were observed between the two deduced peptides, and the expression pattern of Six3 protein in dragon eye is quite similar to common eye during embryogenesis, but from 2 days after hatching, ectopic Six3 expression began to occur in the dragon eye, especially in the outer nuclear layer cells. With eye development, more predominant Six3 distribution was detected in the outer nuclear layer cells of dragon eye than that of normal eye, and fewer cell-layers in outer nuclear layer were observed in dragon eye retina than in normal eye retina. The highlight of this study is that higher Six3 expression occurs in dragon eye goldfish than in normal eye goldfish during retinal development of larvae. (C) 2007 Elsevier Inc. All rights reserved.
A new fluorescent quantitative PCR-based in vitro neutralization assay for white spot syndrome virus
Resumo:
A fluorescent quantitative PCR (FQ-PCR) assay utilizing SYBR green I dye is described for quantitation of white spot syndrome virus (WSSV) particles isolated from infected crayfish, Cambarus clarkii. For this assay, a primer set was designed which amplifies, with high efficiency and specificity, a 129 bp target sequence within ORF167 of the WSSV genome. Conveniently, WSSV particles can be added into the FQ-PCR assay with a simple and convenient method to release its DNA. To establish the basis for an in vitro neutralization test, primary cultures of shrimp cells were challenged with WSSV that had been incubated with a polyclonal anti-WSSV serum or with control proteins. The number of WSSV particles released from the cells after these treatments were assayed by FQ-PCR. This test may serve as a method to screen monoclonal antibody pools or recombinant antibody pools for neutralizing activity prior to in vivo animal experiments. (c) 2007 Elsevier B.V. All rights reserved.
Resumo:
Immunostimulants are the substances, which enhance the non-specific defence mechanism and provide resistance against the invading pathogenic micro-organism. In order to increase the immunity of shrimps against the WSSV, the methanolic extracts of five different herbal medicinal plants like Cyanodon dactylon, Aegle marmelos, Tinospora cordifolia, Picrorhiza kurooa and Eclipta alba were selected and mixed thoroughly in equal proportion. The mixed extract was supplemented with various concentrations viz. 100 (A), 200 (B), 400 (C), and 800 (D) mg kg(-1) through artificial diets individually. The prepared diets (A-D) were fed individually to WSSV free healthy shrimp Penaeus monodon with an average weight of 8.0 +/- 0.5 g for 25 days. Control diet (E), devoid of herbal extract was also fed to shrimps simultaneously. After 25 days of feeding experiment, the shrimps were challenged with WSSV, which were isolated and propagated from the infected crustaceans. The shrimps succumbed to death within 7 days when fed on no herbal immunostimulant diet (E). Among the different concentrations of herbal immunostimulant supplemented diets, the shrimps fed on diet D (800 mg kg(-1)) significantly (P < 0.0001) had more survival (74%) and reduction in the viral load. Also the better performance of haematological, biochemical and immunological parameters was found in the immunostimulant incorporated diets fed shrimps. The present work revealed that the application of herbal immunostimulants will be effective against shrimp viral pathogenesis and they can be recommended for shrimp culture. (c) 2006 Published by Elsevier Ltd.
Resumo:
An antibody phage display library against White Spot Syndrome Virus (WSSV) was constructed. After four rounds of panning against WSSV, 192 out of 480 clones displayed WSSV binding activity. One of the positive clones, designated A1, had relatively higher activity specifically binding to WSSV A1-soluble, single-chain fragment variable (scFv) antibody has an affinity constant (K-aff) of 2.02 +/- 0.42 x 10(9) M-1. Dot blot assays showed that A1-soluble scFv could detect WSSV directly from shrimp hemolymph after 24-h feeding infection by WSSV. A1 scFv has potential for the development of a cheap, simple and sensitive diagnostic kit for WSSV in the field. (C) 2003 Elsevier Science B.V. All rights reserved.
Resumo:
Three virus isolates, RGV-9506, RGV-9807 and RGV-9808, were obtained from cultured pig frogs Rana grylio undergoing lethal infections. Previously, the first isolate, RGV-9506, was shown to be an iridovirus based on ultrastructural and morphological studies. In the present study, the original isolate, along with 2 recent ones, were more extensively characterized by experimental infection studies, histopathology, electron microscopy, serological reactivity, gel electrophoresis of viral polypeptides and DNA restriction fragments, PCR amplification, and nucleic acid sequence analysis of the major capsid protein (MCP) gene. The 3 isolates were shown to be identical to each other, and very similar to FV3, the type species of the genus Ranavirus (family Iridoviridae). These results suggest that RGV should be considered a strain of FV3, and indicate that FV3-like iridoviruses are capable of causing widespread, severe disease among cultured frogs.
Resumo:
Penaeid shrimp is the natural host of White Spot Syndrome Virus (WSSV) that can cause high mortality in the infected hosts. Attempts to obtain sufficient amounts of purified intact WSSV for characterization have been unsuccessful. Using crayfish, Cambarus clarkii as a proliferation system, a large amount of infectious WSSV was reproduced and intact WSSV viral particles were purified with a new isolation medium by ultra-centrifugation. Purified WSSV particles were very sensitive to organic solvents and the detergent, Triton X-100. The size of the rod-shape, somewhat elliptical, intact WSSV was 110-130 x 260-350 mm with a long, tail-like envelope extension. The naked viral nucleocapsid was about 80 x 350 nm, and it possessed 15 spiral and cylindrical helices composed of 14 globular capsomers along its long axis, and a 'ring' structure at one terminus. Distinct WSSV genome DNA patterns were obtained when the purified genomic dsDNA of WSSV was digested with five different restriction enzymes (HindIII, XhoI, B(BamHI, SalI, and SacI). In addition, at least 13 major and distinct protein bands could be observed when purified intact WSSV viruses were separated by SDS-PAGE followed by Coomassie Brilliant R-250 staining. The estimated molecular weights of these proteins were 190, 84, 75, 69, 68, 58, 52, 44, 28, 27.5, 23, 19, and 16 kD, respectively. Both the 44 and 190 kD proteins were easily removed if the hemolymph from the: WSSV infected crayfish was transiently treated with 1%, Triton X-100 before it was subjected to gradient centrifugation, indicating that both of them are located on the surface of the viral envelope. These characteristics are consistent with WSSV isolated from the penaeid shrimp. (C) 2001 Elsevier Science B.V. All rights reserved.
Resumo:
We present a novel high-energy, single-mode, all-fiber-based master-oscillator-power-amplifier (MOPA) laser system operating in the C-band with 3.3-ns pulses and a very widely tunable repetition rate, ranging from 30 kHz to 50 MHz. The laser with a maximum pulse energy of 25 mu J and a repetition rate of 30 kHz is obtained at, a wavelength of 1548 nm by using a double-clad, single-mode, Er:Yb co-doped fiber power amplifier.
Resumo:
利用不同pH值的酸碱溶液处理蚕豆或者离体蚕豆叶片,研究酸碱胁迫下蚕豆叶片保护酶活性的变化。结果表明,离体蚕豆叶片处理10 min和30 min的SOD活性相差不大,整株处理5 h的蚕豆叶片SOD活性远大于处理12 h的SOD活性;离体蚕豆叶片和整株蚕豆处理,较长时间处理下的CAT活性基本上高于短时间处理的CAT活性,短时间处理下(10 min、5 h)POD活性均显著低于对照值,较长时间处理(30 min、12h)下POD活性变化的规律性不强,且变化幅度较小。
Resumo:
非晶状体βγ-晶状体蛋白(non-lens βγ-crystallin, α-亚基)和三叶因子(trefoil factor, β-亚基) 复合物(non-lens βγ-crystallin and trefoil factor complex, 缩写为βγ-CAT)是从大蹼铃蟾皮肤中分离、纯化的一种新型蛋白, 具有促进细胞迁移、伤口愈合功能,同时还可通过核定位、调节转录因子和炎症相关蛋白诱导细胞脱落、凋亡。βγ-CAT对心血管和血液系统的作用和机制还不清楚。本研究的目的在于深入研究这些问题,为人类重大疾病的研究提供新思路。 首先,我们利用各种整体动物模型,研究了βγ-CAT对心血管系统和血液系统的影响。结果发现βγ-CAT可对白细胞、红细胞、血小板、肝细胞、肾细胞和心血管系统产生毒理作用。βγ-CAT可导致白细胞计数、红细胞计数和血小板计数减少、低血压、心律失常、心肌细胞轻度水肿、部分肺泡淤血、炎症细胞浸润肺泡壁、肝细胞水样变性、肾小管水肿、肾小球淤血和脾脏淤血、高钾血症、血糖升高、转氨酶和乳酸脱氢酶升高。我们推测βγ-CAT造成实验动物死亡的主要原因是心功能衰竭、高钾血症和白细胞毒素效应。 其次,为研究βγ-CAT对血管的效应,我们用兔胸主动脉进行了一系列的实验。结果表明,βγ-CAT可引起兔胸主动脉环剂量依赖性收缩,半数有效浓度为(EC50)10 nM; α-肾上腺素能受体阻断剂(酚妥拉明)和5-羟色胺受体阻断剂(S006)不能抑制βγ-CAT的血管收缩效应。因此,我们认为,整体实验观察到的低血压不是由于血管舒张造成的,而是由于βγ-CAT对心肌了产生了抑制效应,并且,βγ-CAT产生的血管收缩效应是通过新的途径引起的。 最后,我们把βγ-CAT导致兔死亡的原因归结为心血管系统衰竭,因为,一方面βγ-CAT抑制心肌使每搏量减少,另一方面它可收缩动脉使心脏的后负荷增加,从而导致重要器官、组织灌注不良而死亡。但导致心功能衰竭的机制还不清楚,于是,我们进行了离体心脏灌流、内皮细胞培养、细胞因子测定、免疫组化、凋亡实验,试图阐明导致动物心力衰竭的机制。首先,我们在离体心脏灌流装置上以恒压和恒流的灌流模式来研究βγ-CAT的心肌变力效应。接下来,我们用高钾去除冠脉血管内皮细胞,以评估内皮细胞在βγ-CAT对心肌的直接效应。最后,我们用βγ-CAT刺激培养的心内皮细胞和主动脉内皮细胞,之后检测细胞因子的释放;并用免疫组化的方法定位冠脉内皮和心肌细胞细胞因子的释放和对这些细胞的凋亡效应。实验一的结果表明,βγ-CAT导致心力衰竭的部分原因是由于βγ-CAT引起的冠脉血管收缩造成;实验二的结果表明,βγ-CAT引起的心力衰竭是内皮依赖的。实验三检测到心内皮、主动脉内皮和冠脉内皮释放TNF-α(TNF-α),高浓度的βγ-CAT还可诱导冠脉内皮凋亡,但不引起心肌细胞凋亡。综上所述,我们认为,冠脉内皮在βγ-CAT引起的心力衰竭中具有较大的贡献,通过冠脉内皮释放细胞因子(如TNF-α)作用于心肌细胞,从而导致收缩能力降低,引起心力衰竭。
Resumo:
To study the influence of Hypericum perforatum extract (HPE) on piglets infected with porcine respiratory and reproductive syndrome virus (PRRSV), enzyme-labeled immunosorbent assay (ELISA) and cytopathic effect (CPE) were used to determine in vitro whether HPE could induce swine pulmonary alveolar macrophages (PAMs) to secrete IFN-gamma, and whether PRRSV titers in PAMs were affected by the levels of HPE-induced IFN-gamma. HPE (200 mg kg(-1)) was administrated by oral gavage to piglets infected with the PRRSV in vivo to observe whether HPE affected the viremia, lung viral titers, and weight gain of piglets infected with PRRSV. The results showed that HPE was capable of inducing PAMs to produce IFN-gamma in a dose dependent manner and HPE pretreatment was capable of significantly reducing PRRSV viral titers in PAMs (P<0.01). Administration of HPE to the PRRSV-infected animals significantly (P<0.05) reduced viremia over time as compared with the PRRSV-infected animals. But there was not significant decrease in lung viral titers at day 21 post-infection between the HPE-treated animals and the PRRSV-infected control piglets. There were no significant differences in weight gain over time among the HPE-treatment animals, the normal control, and the HPE control animals. The PRRSV-infected animals caused significant (P<0.01) growth retardation as compared with the HPE controls and the normal piglets. It suggested that HPE might be an effective novel therapeutic approach to diminish the PRRSV-induced disease in swine.