Apoptosis of human lymphocytes after exposure to hydatid fluid

Modulation of the immune response is an important strategy by which establishment and growth of hydatid cyst in the internal organs of human is warranted. Induction of apoptosis in the lymphocytes might be a considerable component. This study was designed to evaluate apoptotic impact of hydatid fluid [HF] on human lymphocytes. Human lymphocytes were treated with hydatid fluid. After 6 hours of exposure, caspase-3 activity, the central enzyme of apoptosis cascade, was measured by fluorometric assay in the HF-treated lymphocytes and control cells. In addition, the expression of Bax [a pro-apoptotic protein] and Bcl-2 [an anti-apoptotic protein] mRNA was assessed by RT-PCR after 12 hours of exposure. Both the ratio of Bax/Bcl-2 mRNA expression and Caspase-3 activity were higher in the HF-treated lymphocytes relative to the control group. Apoptosis could be as a possible mechanism by which Echinococcus granulosus overwhelms host defenses

Isolation, cloning and expression of the brucella melitensis Omp31 gene

Brucellosis is a zoonotic disease transmitted to humans either from animals or from their products. Although brucellosis can be found worldwide, the Mediterranean Basin, South and Central America, Eastern Europe, Asia, Africa, the Caribbean, and the Middle East have currently been listed as high-risk regions. The genus Brucella is classified in at least nine species. Brucella melitensis is the global pathogenic species of Brucella. The outer membrane protein 31, [Omp31] from B. melitensis is considered as a protective immunogen and an important candidate vaccine. Contamination of purified Omp31 protein by biochemical methods has made some restrictions in practical experiments. In this study, the Omp31 coding gene of B. melitensis Rev 1 strain was inserted in pET32b[+] plasmid with extra His-tag sequence. The integrity of the constructed plasmid was confirmed using restriction enzyme mapping and sequencing. Omp31 was expressed after induction with IPTG in Escherichia coli BL21. Recombinant Omp31 [rOmp31] was purified immunoblotting confirmed immunereactivity of rOmp31. Obtained rOmp31 could be useful as a research experimental tool in protection assays to find its potential as a vaccine candidate

Characterization of Bacillus anthracis spores isolates from soil by biochemical and multiplex PCR analysis

Outbreaks of Bacillus anthracis in animals are repeatedly reported in the Islamic Republic of Iran. In this study soil samples were analysed from endemic regions of the country, and B. anthracis isolates were identified by classical bacteriological and biochemical methods. A multiplex polymerase chain reaction [PCR] assay was also developed as an alternative for identification of isolates, and was shown to be a rapid, sensitive and specific diagnostic assay. The results confirmed that 25 samples contained B. anthracis, of which 9 were virulent for mice and guinea pigs. This study suggests that multiplex PCR can be used as a reliable alternative for the detection of B. anthracis spores

[Development of an indirect ELISA for detection of specific IgG subclasses for saffron pollen allergens]

Allergic reactions to plant pollens are a common problem. There are some evidences that saffron pollen has allergenic properties. Although IgE-mediated hypersensitivity has been reported, other immunologic mechanisms may be involved based on a variety of clinical manifestations in allergic cases. One of the mechanisms that have been suggested is specific IgG and its subclasses. In this study, an Enzyme-linked Immunosorbent Assay [ELISA] was developed for detection of specific IgG subclasses against saffron pollen extract. Briefly, microtiter plates were coated with crude extract of saffron pollen and then blocked with bovine serum albumin [BSA], and the sera were added to the plates which followed by the addition of anti-human IgG. TMB was used as substrate. Finally, after stopping the reaction with addition of HCl, plates read at 450 nm. The method was verified by testing two groups of samples, first, sera from individuals living near saffron farms in Khorasan and second, sera from individuals living in areas not having any saffron farms. The results indicated significant differences between the two groups