Rapid detection of virulent protease secreted by Vibrio anguillarum by dot enzyme-linked immunosorbent assay

Dot enzyme-linked immunosorbent assay (dot-ELISA), indirect ELISA and Western blot were performed to detect the virulent protease secreted by Vibrio anguillarum which was isolated from the diseased left-eyed flounder, Paralichthys olivaceous. Sensitivity results showed that dot-ELISA is a more sensitive, rapid and simple technique for the protease detection. The minimal detectable amount of protease is about 7 pg in the dot-ELISA test, while 7.8 ng in the indirect ELISA and 6.25 ng in the Western blot respectively. Protease could be detected 2 h after incubation of V. anguillarum in the 2216E liquid medium but enzyme activity was very low at that period. From 6 to 12 h, the amount and enzyme activity of protease increased markedly and reached maximum at stationary phase. Analysis of serum samples periodically collected from the infected flounders showed that after 2 h of infection by V. anguillarum, the pathogenic bacteria could be detected in the blood of the infected flounders but no protease was found. It was 5 similar to 6 h after infection that the protease was detected in blood and then the amount increased as infection advanced. Quantitative detection of protease either incubation in the medium or from the blood of infected flounders could be accomplished in virtue of positive controls of quantificational protease standards ("marker") so that the alterations of protease secretion both in vitro and in vivo could be understood generally. In addition, the indirect ELISA and dot-ELISA were also performed to detect V. anguillarum cells. Results indicated that the sensitivity of indirect ELISA to bacteria cells is higher than that of the dot-ELISA, and that the minimal detectable amount is approximately 10(4) cell/mL in the indirect ELISA, while 10(5) cell/mL in the dot-ELISA.

Analysis of three marine fish cell lines by RAPD assay

We tested the applicability of the random amplified polymorphic deoxyribonucleic acid (RAPD) analysis for identification of three marine fish cell lines FG. SPH, and RSBK and as a possible tool to detect cross-contamination. Sixth commercial 10-mer RAPD primers were tested on the cell lines and on samples collected from individual fish. The results obtained showed that the cell lines could be identified to the correspondent species on the basis of identical Patterns produced by 35-48% of the primers tested. the total mean similarity indices for cell lines versus correspondent species of individual fish ranged from 0.825 to 0.851. indicating the existence of genetic variation in these cell lines in relation to the species of their origin. Also, four primers, which gave a monomorphic hand pattern within species/line, but different among the species/line, were obtained. These primers can be useful for identification of these cell lines and for characterization of the genetic variation of these cell lines in relation to the species of their origin. This supported the use of RAPD analysis as an effective tool in species identification and cross-contamination test among different cell lines.

Protein phosphatase inhibition assay for detection of diarrhetic shellfish poison in oyster

A method based on protein phosphatase enzyme activity inhibition for the detection of diarrhetic shellfish poison (DSP) was used to analyze the DSP toxicity in three oyster samples. Based on the standard dose-effect curve developed with a series of okadaic acid (OA) standard solutions, the DSP toxicity of the three oyster samples collected were screened, and the results showed that there were no OA and dinophysis toxins ( DTXs) in the samples without hydrolization. However, the OA toxicity could be detected in two of the hydrolyzed samples, and the OA toxicity of the two samples were 1.81 and 1.21 mu g OA eq./kg oyster, respectively.

Monitoring microbial populations of sulfate-reducing bacteria using an impedimetric immunosensor based on agglutination assay

An impedimetric immunosensor was fabricated for rapid and non-labeled detection of sulfate-reducing bacteria, Desulforibrio caledoiensis (SRB) by immobilizing lectin-Concanavalin A using an agglutination assay. The immobilization of lectin was conducted using amine coupling on the surface of a gold (Au) electrode assembled with 11-Mercaptounclecanoic acid. Electrochemical impedance spectroscopy (EIS) was used to verify the stepwise assembly of the sensor system. The work conditions of the impedimetric immunosensor, such as pH of the buffer solutions and the incubation time of lectin, were optimized. Faradic impedance spectra for charge transfer for the redox probe Fe(CN)(6)(3-/4-) were measured to determine SRB concentrations. The diameter of the Nyquist diagram that is equal to the charge-transfer resistance (RI) increased with increasing SRB concentration. A linear relationship between R-ct and SRB concentration was obtained in SRB concentration range of 1.8 to 1.8 x 10(7) cfu/ml. The variation of the SRB population during the growth process was also monitored using the impedimetric immunosensor. This approach has great potential for simple, low-cost. and time-saving monitoring of microbial populations. (C) 2009 Elsevier B.V. All rights reserved.

基于MIT规则的自适应Unscented卡尔曼滤波及其在旋翼飞行机器人容错控制的应用

提出一种新颖的基于MIT规则的自适应Unscented卡尔曼滤波(Unscented Kalman filter,UKF)算法,用来进行参数以及状态的联合估计。针对旋翼飞行机器人执行器提出一种执行器健康因子(Actuator health coefficients,AHCs)的故障模型结构,应用自适应UKF对AHCs参数进行在线估计,将联合估计的状态以及故障参数引入基于模型的反馈线性化控制结构,组成完整的容错控制系统。提出的自适应UKF算法以及容错控制结构经过中科院沈阳自动化研究所ServoHeli-20旋翼无人智能平台数学模型进行仿真试验验证,效果良好。

MIT SchMUSE: Class-Based Remote Delegation in a Capricious Distributed Environment

MIT SchMUSE (pronounced "shmooz") is a concurrent, distributed, delegation-based object-oriented interactive environment with persistent storage. It is designed to run in a "capricious" network environment, where servers can migrate from site to site and can regularly become unavailable. Our design introduces a new form of unique identifiers called "globally unique tickets" that provide globally unique time/space stamps for objects and classes without being location specific. Object location is achieved by a distributed hierarchical lazy lookup mechanism that we call "realm resolution." We also introduce a novel mechanism called "message deferral" for enhanced reliability in the face of remote delegation. We conclude with a comparison to related work and a projection of future work on MIT SchMUSE.

MIT Scheme Reference Manual

MIT Scheme is an implementation of the Scheme programming language that runs on many popular workstations. The MIT Scheme Reference Manual describes the special forms, procedures, and datatypes provided by the implementation for use by application programmers.

Danieli Hodrovej mit i poetyka miasta

The author analyses the theoretical and historical work of the Czech literary theoretician entitled “Citlivé město (eseje z mytopoetiky”), Praha 2006, ss. 416. Hodrova’s work is focused on the topic of the city as an architectural creation and a complicated social and cultural phenomenon. In the post-modern anthropological thought the city is the subject of interest as an area that defines the identity of the human being through urban notions. The historical model of Hodrova’s reflection is the Czech Prague seen through the eyes of writers, artists, architects but also through the European tradition of presenting the city. Ipso facto, the monograph that reconstructs the artistic means of presenting Prague transforms into a dispute in the area of social psychology, mythical studies and anthropology.

Die Organisation und Leitung der Arbeit Mit Audiovisuellen Lehr- und Lernmitteln an der Humboldt-Universität zu Berlin und an Anderen Hochschulen in der Ddr

Heun Hans-Georg, Die Organisation und Ieitung der Arbeit mit audiovisuellen lehr- und Ißrnmitteln an der Humboldt-Universität zu Berlin und an anderen Hochschulen in der DDR (Organization and conducting of work with audiovidual aids at the Humboldt University and other schools of higher education in the German Democratic Republic), „Neodidagmata” XX, Poznan 1991, Adam Mickiewicz University Press, pp. 131-136. ISBN 83-232-0302-4. ISBN 0077-653X. Received: November 1987. Audiovisual aids have become now an indispensable help for teachers. The centre of audiovisual aids at the Humboldt University in Berlin performs the following tasks: a) production of audiovisual materials; b) technical service of classes and lectures and seminars and repairing of the equipment; c) information on new audiovisual aids and lending services; d) training of students and university teachers in operating audiovisual aids; e) studies on application of audiovisual aids in schools of higher education.

Mit Ofelii we francuskojęzycznej literaturze belgijskiej doby symbolizmu.

Wydział Neofilologia: Instytut Filologii Romańskiej

Evaluation of a WHO-Validated Serotype-Specific Serological Assay for the Diagnosis of Pneumococcal Etiology in Children with Community-Acquired Pneumonia.

BACKGROUND: The etiologic diagnosis of community-acquired pneumonia (CAP) remains challenging in children because blood cultures have low sensitivity. Novel approaches are needed to confirm the role of Streptococcus pneumoniae. METHODS: In this study, pneumococcal aetiology was determined by serology using a subset of blood samples collected during a prospective multicentre observational study of children <15 years of age hospitalised in Belgium with X-ray-confirmed CAP. Blood samples were collected at admission and 3-4 weeks later. Pneumococcal (P)-CAP was defined in the presence of a positive blood or pleural fluid culture. Serotyping of Streptococcus pneumoniae isolates was done with the Quellung reaction. Serological diagnosis was assessed for nine serotypes using World Health Organization validated IgG and IgA serotype-specific enzyme-linked immunosorbent assays (ELISAs). RESULTS: Paired admission/convalescent sera from 163 children were evaluated by ELISA (35 with proven P-CAP and 128 with non proven P-CAP). ELISA detected pneumococci in 82.8% of patients with proven P-CAP. The serotypes identified were the same as with the Quellung reaction in 82% and 59% of cases by IgG ELISA and IgA ELISA, respectively. Overall, ELISA identified a pneumococcal aetiology in 55% of patients with non-proven P-CAP. Serotypes 1 (51.6%), 7F (19%), and 5 (15.7%) were the most frequent according to IgG ELISA. CONCLUSIONS: In conclusion, the serological assay allows recognition of pneumococcal origin in 55% of CAP patients with negative culture. This assay should improve the diagnosis of P-CAP in children and could be a useful tool for future epidemiological studies on childhood CAP etiology.