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BackgroundBrucellosis, as a serious zoonotic infectious disease, has been recognized as a re-emerging disease in the developing countries worldwide. In china, the incidence of brucellosis is increasing each year, seriously threatening the health of humans as well as animal populations. Despite a quite number of diagnostic methods currently being used for brucellosis, innovative technologies are still needed for its rapid and accurate diagnosis, especially in area where traditional diagnostic is unavailable.Methodology/principal findingsIn this study, a total of 22 B cell linear epitopes were predicted from five Brucella outer membrane proteins (OMPs) using an immunoinformatic approach. These epitopes were then chemically synthesized, and with the method of indirect ELISA (iELISA), each of them displayed a certain degree of capability in identifying human brucellosis positive sera. Subsequently, a fusion protein consisting of the 22 predicted epitopes was prokaryotically expressed and used as diagnostic antigen in a newly established brucellosis testing method, nano-ZnO modified paper-based ELISA (nano-p-ELISA). According to the verifying test using a collection of sera collected from brucellosis and non-brucellosis patients, the sensitivity and specificity of multiepitope based nano-p-ELISA were 92.38% and 98.35% respectively. The positive predictive value was 98.26% and the negative predictive value was 91.67%. The multiepitope based fusion protein also displayed significantly higher specificity than Brucella lipopolysaccharide (LPS) antigen.ConclusionsB cell epitopes are important candidates for serologically testing brucellosis. Multiepitope fusion protein based nano-p-ELISA displayed significantly sensitivity and specificity compared to Brucella LPS antigen. The strategy applied in this study will be helpful to develop rapid and accurate diagnostic method for brucellosis in human as well as animal populations.

In recent years, the incidence of brucellosis has increased annually, causing tremendous economic losses to animal husbandry in a lot of countries. Therefore, developing rapid, sensitive, and specific diagnostic techniques is critical to control the spread of brucellosis. In this study, bioinformatics technology was used to predict the B cell epitopes of the main outer membrane proteins of Brucella, and the diagnostic efficacy of each epitope was verified by an indirect enzyme-linked immunosorbent assay (iELISA). Then, a fusion protein containing 22 verified epitopes was prokaryotically expressed and used as an antigen in paper-based ELISA (p-ELISA) for serodiagnosis of brucellosis. The multi-epitope-based p-ELISA was evaluated using a collection of brucellosis-positive and -negative sera collected from bovine and goat, respectively. Receiver operating characteristic (ROC) curve analysis showed that the sensitivity and specificity of detection-ELISA in diagnosing goat brucellosis were 98.85 and 98.51%. The positive and the negative predictive values were 99.29 and 98.15%, respectively. In diagnosing bovine brucellosis, the sensitivity and specificity of this method were 97.85 and 96.61%, with the positive and negative predictive values being identified as 98.28 and 97.33%, respectively. This study demonstrated that the B cell epitopes contained in major antigenic proteins of Brucella can be a very useful antigen source in developing a highly sensitive and specific method for serodiagnosis of brucellosis.

This study was designed to investigate the cytotoxicity of lead acetate (Pb(AC)2, a representative air pollutant) by focusing on PPARγ/caspase-3/PARP apoptotic signaling pathway and to explore the inhibitory effect of PPARγ antagonist on apoptosis of TM3 Leydig cells. MTT assay was utilized to examine cell viability. Cell apoptosis was analyzed using a flow cytometry by staining with Annexin V-PE/7AAD staining and a fluorescence microscope by staining with Hoechst 33,258. The levels of apoptosis-related proteins were examined using western blot. From the results, Pb reduced significantly TM3 cell proliferation in concentration- and time-dependent manner. It increased significantly apoptosis; increased the PPARγ, Bax, procaspase-3, cleaved caspase-3, proPARP, cleaved PARP levels; and decreased Bcl-2 level in Pb-treated TM3 cells as compared to control cells. Furthermore, pretreatment with PPARγ antagonist significantly attenuated the apoptosis and cleavage of caspase-3 and PARP induced by Pb. Our results suggested that Pb induced cytotoxicity on TM3 Leydig cells, at least in part, by increasing PPARγ expression, stimulating cleavage of caspase-3 and PARP, and then induced cell apoptosis. [ABSTRACT FROM AUTHOR]

The multiple ecological niche of one-species (variety) (MENOS) population is formed by one species (variety), but the ecological units have different temporal, spatial, nutritional, temperature, or water ecological niche. The formation of MENOS population is characterized by its specific object, one species (variety), multiple layer canopy, human being's active adjustment, specific niche breadth, and overlap and separation of ecological units. This paper analyzed the characteristics of the super high yield cotton population with dual ecological niche of one-species (variety) (DENOS). As for the yield components, MENOS has the characteristics of high yield canopy and different yield plants.