Tribulus terrestris (Linn.) Attenuates Cellular Alterations Induced by Ischemia in H9c2 Cells Via Antioxidant Potential

Tribulus terrestris L. was evaluated for its cardioprotective property against myocardial ischemia in a cell line model. Initially, methanolic extract was prepared and subjected to sequential extraction with various solvents. The extract with high phenolic content (T. terrestris L. ethyl acetate extract–TTME) was further characterized for its chemical constituents and taken forward for evaluation against cardiac ischemia. HPLC analysis revealed the presence of phenolic compounds like caffeic acid (12.41 ± 0.22 mg g^?1), chlorogenic acid (0.52 ± 0.06 mg g^?1) and 4‐hydroxybenzoic acid (0.60 ± 0.08 mg g^?1). H9c2 cells were pretreated with TTME (10, 25, 50 and 100 µg/ml) for 24 h before the induction of ischemia. Then ischemia was induced by exposing cells to ischemia buffer, in a hypoxic chamber, maintained at 0.1% O₂, 95% N₂ and 5% CO₂, for 1 h. A significant (p ≤ 0.05) increase in reactive oxygen species generation (56%), superoxide production (18%), loss of plasma membrane integrity, dissipation of transmembrane potential, permeability transition pore opening and apoptosis had been observed during ischemia. However, pretreatment with TTME was found to significantly (p ≤ 0.05) attenuate the alterations caused by ischemia. The overall results of this study partially reveal the scientific basis of the use of T. terrestris L. in the traditional system of medicine for heart diseases. Copyright © 2015 John Wiley & Sons, Ltd.     

The bortezomib/proteasome inhibitor PS-341 and triterpenoid CDDO-Im induce synergistic anti-multiple myeloma (MM) activity and overcome bortezomib resistance

The synthetic triterpenoid 2-cyano-3, 12-dioxooleana-1, 9-dien-28-oic acid (CDDO) induces apoptosis in leukemic cells. Here we show that CDDO and its new derivative CDDO-imidazolide (CDDO-Im) trigger apoptosis in multiple myeloma (MM) cells resistant to conventional therapies including melphalan (LR-5), doxorubicin (Dox-40), and dexamethasone (MM.1R, U266, RPMI 8226) without affecting the viability of normal cells. CDDO-IM also triggers apoptosis in bone marrow stromal cells (BMSCs) and decreases interleukin-6 (IL-6) secretion induced by MM cell adhesion to BMSCs. Moreover, CDDO-Im-induced apoptosis in MM cells is not blocked by IL-6 or insulin growth factor-1 (IGF-1). Importantly, CDDO-Im and bortezomib/proteasome inhibitor PS-341 trigger synergistic apoptosis in MM cells associated with loss of mitochondrial membrane potential, superoxide generation, release of mitochondrial proteins cytochrome c/second mitochondria-derived activator of caspases (cytochrome c/Smac), and activation of caspase-8, -9, and -3. Conversely, the pancaspase inhibitor Z-VAD-fmk abrogates the CDDO-Im + bortezomib-induced apoptosis. Low doses of CDDO-Im and bortezomib overcome the cytoprotective effects of antiapoptotic proteins Bcl2 and heat shock protein-27 (Hsp27) as well as nuclear factor-kappa B (NF-kappaB)-mediated growth/survival and drug resistance. Finally, combining CDDO-Im and bortezomib induces apoptosis even in bortezomib-resistant MM patient cells. Together, these findings provide the framework for clinical evaluation of CDDO-Im, either alone or in combination with bortezomib, to overcome drug resistance and improve patient outcome in MM.     

Identification of laminin-binding motifs of Yersinia pestis plasminogen activator by phage display

Yersinia pestis plasminogen activator (Pla), a surface virulence factor contributes to the highly invasive nature of the pathogen by binding various tissue matrix components. In this study we characterised the laminin-binding site(s) of Pla via phage display and alanine-scanning mutagenesis. Previously we isolated 18 different heptamer peptide sequences from a phage display library with biopanning on laminin, and have shown that two of them with sequences of WSLLTPA or YPYIPTL completely inhibited laminin binding of a Pla-expressing recombinant Escherichia coli strain. These phages themselves strongly bound laminin in an ELISA assay utilising horseradish peroxidase-labelled anti-M 13 antibody. In the present study, with the application of synthetic peptides, a 55% and a 33% inhibition was demonstrated using WSLLTPA and YPYIPTL, respectively. Peptide pattern alignment showed two homologous regions for WSLLTPA and one for YPYIPTL inside the Pla sequence. Amino acids were changed for alanine in one of the WSLLTPA regions and in the YPYIPTL region in order to prove the role of the LTP/PTL motifs in laminin binding. Of the four constructed mutants, the triple mutant L65A-T66A-L67A in the WSLLTPA region and the double mutant G178A-L179A in the YPYIPTL region decreased the laminin binding capacity of the Pla-expressing recombinant E. coli by about 50%.     

Subendocardial infarction in a young survivor of aluminium phosphide poisoning

Aluminium phosphide is a solid fumigant pesticide and has systemic toxicity due to liberation of phosphine gas. We report a case of aluminium phosphide poisoning with systemic toxicity in a 16-year-old patient who developed previously undocumented complication of subendocardial infarction, with characteristic electrocardiographic changes reverting back to normal after a period of 10 weeks, even though the patient had clinical recovery much earlier.     

Listeria monocytogenes as a vector for tumor-associated antigens for cancer immunotherapy

As a facultative intracellular bacterium, Listeria monocytogenes has adapted to live within the cytosol of the host cell. It is actively taken up by antigen-presenting cells through phagocytosis, and as Listeria survive within these cells, it is an ideal vector for the delivery of antigens to be processed and presented through both the class I and II antigen-processing pathways. Once phagocytosed, Listeria produces virulence factors within the phagolysosome of the host cell, which allows it to break out of this organelle and live in the host cytosol. It is possible that these virulence factors can enhance the immunogenicity of tumor-associated antigens, which are poorly immunogenic. Recent progress in the development of this bacterium as a vaccine vector for tumor-associated antigens is discussed in the context of bacterial vectors in general. In several mouse models, Listeria-based vaccines have been demonstrated to be an effective method of influencing tumor growth and eliciting potent antitumor immune responses. Safety issues and the transition of Listeria into human clinical trials will also be discussed in this review.     

Evaluation of mutagenicity of wastewater in the vicinity of pesticide industry

Pesticide industrial wastewater samples were taken from the Chinhat industrial area nearby Lucknow city, India. GC–MS analysis revealed the presence of pesticides lindane, α-endosulfan, β-endosulfan, chlorpyriphos, monocrotophos, dimethoate and malathion. A pesticide mixture and wastewater extracts were studied to determine the mutagenicity by Ames Salmonella test, survival of DNA repair defective E. coli K-12 mutants and bacteriophage λ systems. Wastewater samples were concentrated with XAD-resins as an adsorbent and liquid–liquid extraction procedure. The XAD concentrated sample exhibited maximum mutagenic activity in comparison to liquid–liquid extracted sample. TA98 strain was the most responsive strain for both test samples with (+S9) and without (?S9) metabolic activation, while other strains exhibited weak response. A significant decline of DNA repair defective E. coli K-12 mutants, bacteriophage λ was observed with test samples in the survival. The intracellular damage was highest when treated with XAD concentrated sample as compared to liquid–liquid extract after 6 h treatment.     

Diaminothiazoles inhibit angiogenesis efficiently by suppressing Akt phosphorylation

The prevention of neovessel formation or angiogenesis is a recent popular strategy for limiting and curing cancer. Diaminothiazoles are a class of compounds that have been reported to show promise in the treatment of cancer by inhibiting cancer cell proliferation and inducing apoptosis, because of their effects on microtubules and as inhibitors of cyclin-dependent kinases. Many microtubule-targeting agents are being studied for their antiangiogenic activity, and a few have shown promising activity in the treatment of cancer. Here, we report that diaminothiazoles can be highly effective as antiangiogenic agents, as observed in the chick membrane assay. The lead compound, 4-amino-5-benzoyl-2-(4-methoxyphenylamino)thiazole (DAT1), inhibits endothelial cell processes such as invasion, migration, and tubule formation, which require a functional cytoskeleton. DAT1 also decreases the expression of cell adhesion markers. The antiangiogenic activities of DAT1 occur at concentrations that are not cytotoxic to the normal endothelium. Analysis of intracellular signaling pathways shows that DAT1 inhibits Akt phosphorylation, which is actively involved in the angiogenic process. The antiangiogenic properties of diaminothiazoles, in addition to their promising antimitotic and cytotoxic properties in cancer cell lines, give them an extra advantage in the treatment of cancer.