Nephrin regulates lamellipodia formation by assembling a protein complex that includes Ship2, filamin and lamellipodin

Actin dynamics has emerged at the forefront of podocyte biology. Slit diaphragm junctional adhesion protein Nephrin is necessary for development of the podocyte morphology and transduces phosphorylation-dependent signals that regulate cytoskeletal dynamics. The present study extends our understanding of Nephrin function by showing in cultured podocytes that Nephrin activation induced actin dynamics is necessary for lamellipodia formation. Upon activation Nephrin recruits and regulates a protein complex that includes Ship2 (SH2 domain containing 5' inositol phosphatase), Filamin and Lamellipodin, proteins important in regulation of actin and focal adhesion dynamics, as well as lamellipodia formation. Using the previously described CD16-Nephrin clustering system, Nephrin ligation or activation resulted in phosphorylation of the actin crosslinking protein Filamin in a p21 activated kinase dependent manner. Nephrin activation in cell culture results in formation of lamellipodia, a process that requires specialized actin dynamics at the leading edge of the cell along with focal adhesion turnover. In the CD16-Nephrin clustering model, Nephrin ligation resulted in abnormal morphology of actin tails in human podocytes when Ship2, Filamin or Lamellipodin were individually knocked down. We also observed decreased lamellipodia formation and cell migration in these knock down cells. These data provide evidence that Nephrin not only initiates actin polymerization but also assembles a protein complex that is necessary to regulate the architecture of the generated actin filament network and focal adhesion dynamics.     

Development of insect resistant transgenic cotton lines expressing cry1EC gene from an insect bite and wound inducible promoter

Transgenic cotton lines were developed for high-level expression of a synthetic cry1EC gene from a wound inducible promoter. The tobacco pathogenesis related promoter PR-1a was modified by placing CaMV35S promoter on its upstream in reverse orientation. The resultant chimeric promoter CaMV35S(r)PR-1a expressed constitutively and was further up-regulated at the site of feeding by insects. It was induced more rapidly by treatment with salicylic acid (SA). The CaMV35S(r)PR-1a cry1EC expressing transgenic lines of cotton showed 100% mortality of Spodoptera litura larvae. The tightly regulated low-level expression of PR-1a was modified to a highly expressing constitutive expression by CaMV35S placed in reverse orientation. Salicylic acid treatment and wounding enhanced the expression further by the chimeric promoter. The leaves expressed more δ-endotoxin around the sites of insect bites. The levels of expression and induction varied among different transgenic lines, suggesting position effect. Some of the transgenic lines that expressed Cry1EC from the chimeric promoter at a low level also showed 100% mortality when induced with salicylic acid. A highly expressing insect bite and wound inducible promoter is desirable for developing insect resistant transgenic plants.     

Low‐intensity microwave irradiation does not substantially alter gene expression in late larval and adult Caenorhabditis elegans

Reports that low‐intensity microwave radiation induces heat‐shock reporter gene expression in the nematode, Caenorhabditis elegans, have recently been reinterpreted as a subtle thermal effect caused by slight heating. This study used a microwave exposure system (1.0 GHz, 0.5 W power input; SAR 0.9–3 mW kg^?1 for 6‐well plates) that minimises temperature differentials between sham and exposed conditions (≤0.1 °C). Parallel measurement and simulation studies of SAR distribution within this exposure system are presented. We compared five Affymetrix gene arrays of pooled triplicate RNA populations from sham‐exposed L4/adult worms against five gene arrays of pooled RNA from microwave‐exposed worms (taken from the same source population in each run). No genes showed consistent expression changes across all five comparisons, and all expression changes appeared modest after normalisation (≤40% up‐ or down‐regulated). The number of statistically significant differences in gene expression (846) was less than the false‐positive rate expected by chance (1131). We conclude that the pattern of gene expression in L4/adult C. elegans is substantially unaffected by low‐intensity microwave radiation; the minor changes observed in this study could well be false positives. As a positive control, we compared RNA samples from N2 worms subjected to a mild heat‐shock treatment (30 °C) against controls at 26 °C (two gene arrays per condition). As expected, heat‐shock genes are strongly up‐regulated at 30 °C, particularly an hsp‐70 family member (C12C8.1) and hsp‐16.2. Under these heat‐shock conditions, we confirmed that an hsp‐16.2::GFP transgene was strongly up‐regulated, whereas two non‐heat‐inducible transgenes (daf‐16::GFP; cyp‐34A9::GFP) showed little change in expression. Bioelectromagnetics 30:602–612, 2009. © 2009 Wiley‐Liss, Inc.     

Studies on Guizotia abyssinica L. oil: Biodiesel synthesis and process optimization

Guizotia abyssinica seeds, a common bird feedstock, have been explored for the potential of biodiesel synthesis. The oil was extracted from the seeds by solvent extraction and composition of G. abyssinica oil was examined. The reaction parameters for biodiesel synthesis have been optimized. Temperature, oil: methanol ratio, catalyst type and catalyst concentration were found to have significant role on ester conversion. According to this study, the maximum yield of ester (98.7%) can be obtained with optimized sodium methoxide catalyst dosage (0.6%) at an operational temperature of 65 °C. Methyl ester of G. abyssinica oil was also studied for its oxidation stability and low temperature properties. Further, the synthesized product was blended in diesel at 5–20% ratios and evaluated for physico-chemical properties.     

Syntheses of novel high affinity ligands for opioid receptors

A series of novel high affinity opioid receptor ligands have been made whereby the phenolic-OH group of nalbuphine, naltrexone methiodide, 6-desoxonaltrexone, hydromorphone and naltrindole was replaced by a carboxamido group and the furan ring was opened to the corresponding 4-OH derivatives. These furan ring ‘open’ derivatives display very high affinity for μ and κ receptors and much less affinity for δ. The observation that these target compounds have much higher receptor affinity than the corresponding ring ‘closed’ carboxamides significantly strengthens our underlying pharmacophore hypothesis concerning the bioactive conformation of the carboxamide group.     

Taxonomic studies on genus Tetramorium Mayr (Hymenoptera, Formicidae) with report of two new species and three new records including a tramp species from India with a revised key

Two new species of Tetramorium Mayr, namely Tetramorium shivalikensesp. n. and Tetramorium triangulatumsp. n. are described. Tetramorium triangulatumsp. n. belongs to the inglebyi-species group and is described based on worker, queen and male caste, while Tetramorium shivalikensesp. n. belongs to the ciliatum-species group and is described based on worker caste only. Three species viz., Tetramorium caldarium (Roger), Tetramorium tonganum Mayr and Tetramorium urbanii Bolton represent first records from India. The male caste is described for the first time in the case of Tetramorium tonganum. Among these, Tetramorium caldarium is a tramp species which extends its limit to India as well. A revised key to the Indian ants of this genus is also provided herewith.     

YspC: A Unique Translocator Exhibits Structural Alteration in the Complex form with Chaperone SycB

YspC is an annotated translocator of Yersinia secretion apparatus-Yersinia secretion protein type three secretion system of Yersinia enterocolitica, it forms an 1:1 complex with its cognate chaperone SycB. Unlike other translocators, YspC is highly soluble inspite of having a transmembrane region. Size exclusion chromatography shows that YspC exists predominantly in a monomeric form. Multiple sequence alignment and ConSurf (a web based bioinformatic tool) analysis confirm its significant deviation from the closest class of minor translocators. YspC also possesses a tertiary structure signal seen from near UV CD, further confirming its unique nature amongst the groups of translocators. Far UV CD depicts that YspC is predominantly an ??-helical protein; however, its secondary structure alters in the YspC-SycB complex. Thermal denaturation curve predicts a cooperative melting behaviour for YspC which is altered in the YspC-SycB complex. Furthermore, trypsinolysis data confirms a different digestion pattern for YspC in isolation, when compared to the complex form with SycB. From the Forsters resonance energy transfer analysis, it can be predicted that the two tetratricopeptide repeat regions of SycB are masked while it forms a complex with YspC and this is further confirmed by the interaction studies of YspC with two truncated forms of SycB. YspC interacted with ?SycB_(1?C114) and ?SycB_(36?C114) (possessing only the two TPR regions). However, the complexes formed between YspC and truncated forms of SycB have altered physiological states.     

Spatial and temporal variation of secondary metabolite profiles in Ginkgo biloba leaves

Terpene trilactones (TTLs) are the main bioactive constituents of Ginkgo biloba used for the preparation of drugs for several ailments. Flavonoid glycosides (FGs) are the significant group of compounds found in Ginkgo leaves used in food and healthcare products. The variation in the content of bioactive constituents, as well as antioxidant activity, with respect to change in altitude and the time of sample collection was investigated. The amount of TTLs varied from 0.218-0.709% w/w, whereas FGs were observed in the range of 0.130-0.209% w/w. Ginkgolides J and A showed significant variation (p ≤ 0.05) with the change in altitude, while other components remained more or less unaffected. The amount of TTLs and FGs were not affected significantly by the time of sample collection. The antioxidant capacity (by using DPPH, ABTS, and FRAP assays) of ginkgo extracts was highly dependent on the altitude, and was at maximum in the samples collected from 1000-2000 m, followed by the samples collected from altitudes lower than 1000 m and higher than 2000 m. Thus, Ginkgo harvested from 1000-2000 m of altitudinal range irrespective of the season could provide better nutraceutical formulations, whereas that from below 1000 m may yield a terpene trilactone-enriched fraction.     

Methane emission potential and increased efficiency of a phytoremediation system bioaugmented with Bacillus firmus XJSL 1-10

Horizontal subsurface flow constructed wetland mesocosms (HSSCW) designed to treat municipal waste water were bioaugmented with Bacillus firmus XJSL 1-10. The efficiencies of the three HSSCW mesocosms (non-vegetated HSSCW, Schoenoplectus validus HSSCW and Bambusa vulgaris HSSCW) were assessed. Bioaugmentation not only enhanced the efficiency of the phytoremediation system but also reduced methane emission from an average of 51.3 mg/m2/d to 21.6 mg/m2/d in Schoenoplectus validus HSSCW and from an average of 1708 mg/m2/d to 1473 mg/m2/d in Bambusa vulgaris HSSCW. Each of the three types of bioaugmented HSSCWs showed higher purification efficiency with respect to the removal of BOD and NH4-N than the non-bioaugmented HSSCWs. The performance enhancement was most significant in bioaugmented Schoenoplectus validus HSSCW mesocosm with 48.8 and 44.8% lower BOD, and NH4-N, respectively than the non-bioaugmented HSSCW.