A sensitive & specific multiplex PCR assay for simultaneous detection of Bacillus anthracis, Yersinia pestis, Burkholderia pseudomallei & Brucella species.

Background & objectives: Bacillus anthracis, Yersinia pestis, Burkholderia pseudomallei and Brucella species are potential biowarfare agents. Classical bacteriological methods for their identification are cumbersome, time consuming and of potential risk to the handler. Methods: We describe a sensitive and specific multiplex polymerase chain reaction (mPCR) assay involving novel primers sets for the simultaneous detection of B. anthracis, Y. pestis, B. pseudomallei and Brucella species. An additional non-competitive internal amplification control (IAC) was also included. Results: The mPCR was found to be specific when tested against closely related organisms. The sensitivity of the assay in spiked blood samples was 50 colony forming units (cfus)/25 μl reaction, for the detection of B. anthracis, Y. pestis and Brucella species; and 150 cfus/25 μl reaction, for B. pseudomallei. The assay proved useful in correctly and promptly identifing the clinical isolates of the targeted agents recovered from patients, compared to the gold standard culture methods. Interpretation & conclusion: The assay described in this study showed promise to be useful in application as a routine detection cum diagnostic method for these pathogens.

Epigenetics and epigenetic mechanisms in disease with emphasis on autoimmune diseases.

Epigenetics deals with molecular heritable patterns relating to chromatin, which exists in two alterable and inter-convertible states. The two conformations of chromatin i.e., compact and relaxed are due to epigenetic regulation. The alterations in chromatin normalize gene expression patterns. Thus, the epigenetic marks on chromatin are the deciding factors for either gene silencing or activation. The epigenetics introduced a new term viz., epiallele which deviates from the classical Mendelian allele. The remodeling of chromatin during embryonic phase, post-translational aberrations of chromatin proteins causing cellular dysfunction and possible epigenetic therapies are discussed in the present article. The role of epigenetic mechanisms in triggering / progression of several autoimmune diseases is being emphasized off late. The study of such complex epigenetic processes becomes very important in understanding the etiopathology of the disease as well as in designing target therapies.

Copper & biological health.

Cu being a transition metal is ubiquitously engaged in biological systems to derive electrons through its participation in several enzymatic reactions. Upon bestowing the significance of Cu in biological systems, an elaborate mechanism is set forth by nature for maintaining Cu homeostasis. As a consequence, a wide variety of proteins viz., family of Cu bearing proteins, cuproenzymes, Cu transporters and Cu chaperone proteins have been manifested for enabling Cu to show its relevance in biological health. In addition, understanding the role of Cu in hepatic and neuronal functions and also in angiogenesis keeps progressing with the advent of novel molecular tools. The studies on genetic defects in Cu metabolism causing abnormalities are providing insights leading to the possible prognostic cues to alleviate the sufferings.