Insight towards the conserved water mediated recognition of catalytic and structural Zn+2 ions in human Matrix Metalloproteinase-8 enzyme: A study by MD-simulation methods

Human matrix metalloproteinase-8 (hMMP-8) plays a important role in the progression of colorectal cancer, metastasis, multiple sclerosis and rheumetoid arthritis. Extensive MD-simulation of the PDB and solvated structures of hMMP-8 has revealed the presence of few conserved water molecules around the catalytic and structural zinc (ZnC and ZnS) ions. The coordination of two conserved water molecules (W and WS) to ZnS and the H-bonding interaction of WS to S151 have indicated the plausible involvement of that metal ion in the catalytic process. Beside this the coupling of ZnC and ZnS metal ions (ZnC – W_H (W₁)…..W₂ ….H₁₆₂ - ZnS) through two conserved hydrophilic centers (occupied by water molecules) may also provide some rational on the recognition of two zinc ions which were separated by ~13 Å in their X-ray structures. This unique recognition of both the Zn⁺² ions in the enzyme through conserved water molecules may be implemented/ exploited for the design of antiproteolytic agent using water mimic drug design protocol.     

Toxicity of cypermethrin: hsp70 as a biomarker of response in transgenic Drosophila

Heat shock protein induction is often associated with a cellular response to a harmful environment or to adverse life conditions. The main aims of our study were (1) to evaluate the cytotoxic potential of cypermethrin; and (2) to investigate the suitability of stress-induced heat shock protein Hsp70 as a biomarker for environmental pollutants in transgenic Drosophila melanogaster (Hsp70-lacZ)Bg⁹. Different concentrations of cypermethrin (0.002, 0.2, 0.5 and 50.0 p.p.m.) were mixed with food. Third instar larvae of transgenic Drosophila melanogaster were allowed to feed on these mixtures for different time intervals (2, 4, 6, 12, 24 and 48h). Following feeding, hsp70 induction and tissue damage were evaluated. In the highest concentration treatment group (50 p.p.m.), 100% larval mortality was recorded after 12 h exposure. Hsp70 was found to be induced even at the lowest concentration (0.002 p.p.m.) of the insecticide, while tissue damage was observed in the larvae exposed for 48 h. While an insignificant decline in hsp70 expression was observed in the larvae exposed to cypermethrin at a dietary concentration of 0.002 p.p.m. after 48 h compared with those exposed for 24 h, in the next two higher concentrations of the toxicant, a similar but significant decline in hsp70 expression was evident in the exposed larvae after 48 h. The present study reveals the cytotoxic potential of cypermethrin and further proposes that hsp70 induction in transgenic Drosophila melanogaster could be used as a sensitive biomarker in risk assessment.     

Systematic Review and Consensus Guidelines for Environmental Sampling of Burkholderia pseudomallei

Background

Burkholderia pseudomallei, a Tier 1 Select Agent and the cause of melioidosis, is a Gram-negative bacillus present in the environment in many tropical countries. Defining the global pattern of B. pseudomallei distribution underpins efforts to prevent infection, and is dependent upon robust environmental sampling methodology. Our objective was to review the literature on the detection of environmental B. pseudomallei, update the risk map for melioidosis, and propose international consensus guidelines for soil sampling.

Methods/Principal Findings

An international working party (Detection of Environmental Burkholderia pseudomallei Working Party (DEBWorP)) was formed during the VIth World Melioidosis Congress in 2010. PubMed (January 1912 to December 2011) was searched using the following MeSH terms: pseudomallei or melioidosis. Bibliographies were hand-searched for secondary references. The reported geographical distribution of B. pseudomallei in the environment was mapped and categorized as definite, probable, or possible. The methodology used for detecting environmental B. pseudomallei was extracted and collated. We found that global coverage was patchy, with a lack of studies in many areas where melioidosis is suspected to occur. The sampling strategies and bacterial identification methods used were highly variable, and not all were robust. We developed consensus guidelines with the goals of reducing the probability of false-negative results, and the provision of affordable and ‘low-tech’ methodology that is applicable in both developed and developing countries.

Conclusions/Significance

The proposed consensus guidelines provide the basis for the development of an accurate and comprehensive global map of environmental B. pseudomallei.     

Systems Model of T Cell Receptor Proximal Signaling Reveals Emergent Ultrasensitivity

Receptor phosphorylation is thought to be tightly regulated because phosphorylated receptors initiate signaling cascades leading to cellular activation. The T cell antigen receptor (TCR) on the surface of T cells is phosphorylated by the kinase Lck and dephosphorylated by the phosphatase CD45 on multiple immunoreceptor tyrosine-based activation motifs (ITAMs). Intriguingly, Lck sequentially phosphorylates ITAMs and ZAP-70, a cytosolic kinase, binds to phosphorylated ITAMs with differential affinities. The purpose of multiple ITAMs, their sequential phosphorylation, and the differential ZAP-70 affinities are unknown. Here, we use a systems model to show that this signaling architecture produces emergent ultrasensitivity resulting in switch-like responses at the scale of individual TCRs. Importantly, this switch-like response is an emergent property, so that removal of multiple ITAMs, sequential phosphorylation, or differential affinities abolishes the switch. We propose that highly regulated TCR phosphorylation is achieved by an emergent switch-like response and use the systems model to design novel chimeric antigen receptors for therapy.     

Variation in the Activity of Three Principal Detoxifying Enzymes in Major Sucking Pest of Tea, Helopeltis theivora Waterhouse (Heteroptera: Miridae) from Sub-Himalayan Tea Plantations of West Bengal, India

Helopeltis theivora Waterhouse (Heteroptera: Miridae), is a major sucking pest of tea in North East India along with other tea growing countries. In West Bengal, tea is cultivated in three sub-Himalayan regions, Terai (foothill plains western to river Teesta), the Dooars (foothill plains eastern to river Teesta) and the Darjeeling hill slopes. Most plantations, in these regions are managed conventionally i.e. by spraying different synthetic insecticides and a few by organic farming using different herbal and microbial insecticides. In conventional plantations, continuous application of insecticides may lead to the selection of more tolerant H. theivora populations making the pest difficult to control. So, there is a pressing need to know the biochemical variability in relation to the metabolic resistance in the pest populations and develop a population specific control strategy. Activity of three principal insecticide detoxifying enzymes in H. theivora populations were studied from three tea growing regions of North Bengal. Higher levels of activity of all the studied enzymes were found from conventional tea plantations. In male H. theivora, the activity of general esterases (GEs) was 6.6–11.2 and 10.5–11.4 fold higher, cytochrome P450 (CYPs) was 2.0–3.2 and 3.0–3.2 fold higher and glutathione S-transferase (GSTs) was 5.2–8.3 and 6.4–8.7 fold higher in Terai and the Dooars populations, respectively than organic populations from Darjeeling hill slopes. Similarly, in female H. theivora, activity of GEs was 6.2–10.3 and 8.3–9.6 folds higher, CYPs was 1.9–3.2 and 3.0–3.3 fold, and GSTs was 3.5–5.4 and 4.4–6.0 fold higher in Terai and the Dooars, respectively than the organic populations from Darjeeling hill slopes. The activities of all three enzymes were found to be significantly low in organic plantations from Darjeeling hill slopes. Esterase I–VI isozymes with higher level of expression were found in specimens from conventional tea plantations than organic populations of H. theivora from Darjeeling hill slopes in isozymes study.     

Habitat Preference and Diversity of Anuran in Durgapur, an Industrial City of West Bengal, India

Anuran diversity study was conducted for one year between June 2008 and August 2009 at Durgapur, an industrial city of West Bengal, India. Nine species under four families of the Order Anura were found to occur in Durgapur. They have a wide range of habitat preference like permanent or temporary aquatic bodies, human residential area, forested areas, termite nest, tree hole, under spaces of logs and so forth. Of the nine sp. 8 were confined to Amarabati pond (site-I), 5 were confined to Durgapur Government College campus (site-II) and 7 were confined at Fuljhore (site-III) of Durgapur. Analysis of the collected data on the anuran community of the study area revealed Shannon–Wiener species diversity index minimum (1.312) at site-II and maximum (1.938) at site-III. In contrast Margalef richness index value was minimum (0.627) at site-II and maximum (1.424) at site-III.     

Regulation of β-catenin stabilization in human platelets

The Wnt/β-catenin pathway controls developmental processes and homeostasis; however, abnormal activation of this pathway has been linked to several human diseases. Recent reports have demonstrated regulation of platelet function by canonical and non-canonical Wnt signalling. Platelet aggregation plays a crucial role in haemostasis and thrombosis. Here we report for the first time that, induction of sustained aggregation of platelets by a strong agonist in the presence of calcium was associated with nearly complete proteolysis of β-catenin, which was abrogated upon depletion of calcium from platelet suspension. β-catenin cleavage was disallowed in absence of aggregation, thus implicating integrin α_IIbβ₃ engagement in β-catenin proteolysis. Degradation of β-catenin was blocked partially by inhibitors of either proteasome or calpain and completely when cells were exposed to both the inhibitors. Protein kinase C inhibition, too, abolished β-catenin degradation. Thus activities of proteasome, calpain and protein kinase C regulate stabilization of β-catenin in aggregated human platelets.