Functional characterization, homology modeling and docking studies of beta-glucosidase responsible for bioactivation of cyanogenic hydroxynitrile glucosides from Leucaena leucocephala (subabul)

Glycosyl hydrolase family 1 beta-glucosidases are important enzymes that serve many diverse functions in plants including defense, whereby hydrolyzing the defensive compounds such as hydroxynitrile glucosides. A hydroxynitrile glucoside cleaving beta-glucosidase gene (Llbglu1) was isolated from Leucaena leucocephala, cloned into pET-28a (+) and expressed in E. coli BL21 (DE3) cells. The recombinant enzyme was purified by Ni-NTA affinity chromatography. The optimal temperature and pH for this beta-glucosidase were found to be 45 A degrees C and 4.8, respectively. The purified Llbglu1 enzyme hydrolyzed the synthetic glycosides, pNPGlucoside (pNPGlc) and pNPGalactoside (pNPGal). Also, the enzyme hydrolyzed amygdalin, a hydroxynitrile glycoside and a few of the tested flavonoid and isoflavonoid glucosides. The kinetic parameters K (m) and V (max) were found to be 38.59 mu M and 0.8237 mu M/mg/min for pNPGlc, whereas for pNPGal the values were observed as 1845 mu M and 0.1037 mu M/mg/min. In the present study, a three dimensional (3D) model of the Llbglu1 was built by MODELLER software to find out the substrate binding sites and the quality of the model was examined using the program PROCHEK. Docking studies indicated that conserved active site residues are Glu 199, Glu 413, His 153, Asn 198, Val 270, Asn 340, and Trp 462. Docking of rhodiocyanoside A with the modeled Llbglu1 resulted in a binding with free energy change (Delta G) of -5.52 kcal/mol on which basis rhodiocyanoside A could be considered as a potential substrate.

On the zeta function of a periodic-finite-type shift

Periodic-finite-type shifts (PFT's) are sofic shifts which forbid the appearance of finitely many pre-specified words in a periodic manner. The class of PFT's strictly includes the class of shifts of finite type (SFT's). The zeta function of a PET is a generating function for the number of periodic sequences in the shift. For a general sofic shift, there exists a formula, attributed to Manning and Bowen, which computes the zeta function of the shift from certain auxiliary graphs constructed from a presentation of the shift. In this paper, we derive an interesting alternative formula computable from certain ``word-based graphs'' constructed from the periodically-forbidden word description of the PET. The advantages of our formula over the Manning-Bowen formula are discussed.

Flexibility in Food Extraction Techniques in Urban Free-Ranging Bonnet Macaques, Macaca radiata

Non-human primate populations, other than responding appropriately to naturally occurring challenges, also need to cope with anthropogenic factors such as environmental pollution, resource depletion, and habitat destruction. Populations and individuals are likely to show considerable variations in food extraction abilities, with some populations and individuals more efficient than others at exploiting a set of resources. In this study, we examined among urban free-ranging bonnet macaques, Macaca radiata (a) local differences in food extraction abilities, (b) between-individual variation and within-individual consistency in problem-solving success and the underlying problem-solving characteristics, and (c) behavioral patterns associated with higher efficiency in food extraction. When presented with novel food extraction tasks, the urban macaques having more frequent exposure to novel physical objects in their surroundings, extracted food material from PET bottles and also solved another food extraction task (i.e., extracting an orange from a wire mesh box), more often than those living under more natural conditions. Adults solved the tasks more frequently than juveniles, and females more frequently than males. Both solution-technique and problem-solving characteristics varied across individuals but remained consistent within each individual across the successive presentations of PET bottles. The macaques that solved the tasks showed lesser within-individual variation in their food extraction behavior as compared to those that failed to solve the tasks. A few macaques appropriately modified their problem-solving behavior in accordance with the task requirements and solved the modified versions of the tasks without trial-and-error learning. These observations are ecologically relevant - they demonstrate considerable local differences in food extraction abilities, between-individual variation and within-individual consistency in food extraction techniques among free-ranging bonnet macaques, possibly affecting the species' local adaptability and resilience to environmental changes.

Nanostructure Patterning on Flexible Substrates Using Electron Beam Lithography

Patterning nanostructures on flexible substrates plays a key role in the emerging flexible electronics technology. The flexible electronic devices are inexpensive and can be conformed to any shape. The potential applications for such devices are sensors, displays, solar cells, RFID, high-density biochips, optoelectronics etc. E-beam lithography is established as a powerful tool for nanoscale fabrication, but its applicability on insulating flexible substrates is often limited because of surface charging effects. This paper presents the fabrication of nanostructures on insulating flexible substrates using low energy E-beam lithography along with metallic layers for charge dissipation. Nano Structures are patterned on different substrates of materials such as acetate and PET foils. The fabrication process parameters such as the proximity gap of exposure, the exposure dosage and developing conditions have been optimized for each substrate.

Polyester derived from recycled poly(ethylene terephthalate) waste for regenerative medicine

Despite advances in regenerative medicine, the cost of such therapies is beyond the reach of many patients globally in part due to the use of expensive biomedical polymers. Large volumes of poly(ethylene terephthalate) (PET) in municipal waste is a potential source of low cost polymers. A novel polyester was prepared by a catalyst-free, melt polycondensation reaction of bis(hydroxyethylene) terephthalate derived from PET post-consumer waste with other multi-functional monomers from renewable sources such as citric acid, sebacic acid and D-mannitol. The mechanical properties and degradation rate of the polyester can be tuned by varying the composition and the post-polymerization time. The polyester was found to be elastomeric, showed excellent cytocompatibility in vitro and elicited minimal immune response in vivo. Three-dimensional porous scaffolds facilitated osteogenic differentiation and mineralization. This class of polyester derived from low cost, recycled waste and renewable sources is a promising candidate for use in regenerative medicine.

Effects of Withania somnifera and Tinospora cordifolia Extracts on the Side Population Phenotype of Human Epithelial Cancer Cells: Toward Targeting Multidrug Resistance in Cancer

Recent reports suggest the existence of a subpopulation of stem-like cancer cells, termed as cancer stem cells (CSCs), which bear functional and phenotypic resemblance with the adult, tissue-resident stem cells. Side population (SP) assay based on differential efflux of Hoechst 33342 has been effectively used for the isolation of CSCs. The drug resistance properties of SP cells are typically due to the increased expression of ABC transporters leading to drug efflux. Conventionally used chemotherapeutic drugs may often leads to an enrichment of SP, revealing their inability to target the drug-resistant SP and CSCs. Thus, identification of agents that can reduce the SP phenotype is currently in vogue in cancer therapeutics. Withania somnifera (WS) and Tinospora cordifolia (TC) have been used in Ayurveda for treating various diseases, including cancer. In the current study, we have investigated the effects of ethanolic (ET) extracts of WS and TC on the cancer SP phenotype. Interestingly, we found significant decrease in SP on treatment with TC-ET, but not with WS-ET. The SP-inhibitory TC-ET was further fractionated into petroleum ether (TC-PET), dichloromethane (TC-DCM), and n-butyl alcohol (TC-nBT) fractions using bioactivity-guided fractionation. Our data revealed that TC-PET and TC-DCM, but not TC-nBT, significantly inhibited SP in a dose-dependent manner. Furthermore, flow cytometry-based functional assays revealed that TC-PET and TC-DCM significantly inhibited ABC-B1 and ABC-G2 transporters and sensitized cancer cells toward chemotherapeutic drug-mediated cytotoxicity. Thus, the TC-PET and TC-DCM may harbor phytochemicals with the potential to reverse the drug-resistant phenotype, thus improving the efficacy of cancer chemotherapy.