Myogenesis defect due to Toca-1 knockdown can be suppressed by expression of N-WASP

Skeletal muscle formation is a multistep process involving proliferation, differentiation, alignment and fusion of myoblasts to form myotubes which fuse with additional myoblast to form myofibers. Toca-1 (Transducer of Cdc42-dependent actin assembly), is an adaptor protein which activates N-WASP in conjunction with Cdc42 to facilitate membrane invagination, endocytosis and actin cytoskeleton remodeling. Expression of Toca-1 in mouse primary myoblasts and C2C12 myoblasts was up-regulated on day 1 of differentiation and subsequently down-regulated during differentiation. Knocking down Toca-1 expression in C2C12 cells (Toca-1^KD cells) resulted in a significant decrease in myotube formation and expression of shRNA-resistant Toca-1 in Toca-1^KD cells rescued the myogenic defect, suggesting that the knockdown was specific and Toca-1 is essential for myotube formation. Toca-1^KD cells exhibited elongated spindle-like morphology, expressed myogenic markers (MyoD and MyHC) and localized N-Cadherin at cell periphery similar to control cells suggesting that Toca-1 is not essential for morphological changes or expression of proteins critical for differentiation. Toca-1^KD cells displayed prominent actin fibers suggesting a defect in actin cytoskeleton turnover necessary for cell–cell fusion. Toca-1^KD cells migrated faster than control cells and had a reduced number of vinculin patches similar to N-WASP^KO MEF cells. Transfection of N-WASP-expressing plasmid into Toca-1^KD cells restored myotube formation of Toca-1^KD cells. Thus, our results suggest that Toca-1^KD cells have defects in formation of myotubes probably due to reduced activity of actin cytoskeleton regulators such as N-WASP. This is the first study to identify and characterize the role of Toca-1 in myogenesis.     

Insecticidal and antifungal potentials of a proteinaceous extract of Cassia occidentalis (Linn.) seeds

Proteinaceous extract obtained from Cassia occidentalis seeds with purification fold of 3.91 and 82.7% of bovine trypsin inhibitory activity was assessed at different concentrations (25, 50, 100, 200, 400 and 800 μg/ml) against Spodoptera litura. Assay of larval feeding suggested proteinaceous extract to be toxic as prepupal (80.16%) and pupal mortalities (100%) along with growth deterrent effect with only 16.71% pupation was observed at 800 μg/ml. Fifty per cent mortality (LC₅₀) was observed at 132.91 μg/ml. Also the inhibitor affected fecundity, longevity and percentage of egg hatching. Nutritional indices were adversely affected as both efficiency of conversion of ingested and digested food decreased while approximate digestibility and metabolic cost increased. In vitro studies on proteolytic enzymes of S. litura revealed inhibition of trypsin and chymotrypsin in lumen and faecal matter at all tested concentrations. Also proteinaceous extract inhibited mycelial growth in Fusarium oxysporum, Alternaria brassicicola and Alternaria alternata at 100 μg/ml.     

Epiphytic succession on tree trunks in a mixed oak-cedar forest,Kumaun Himalaya

The epiphytes present at about breast height on trunks of different size were studied for three major tree species in a seasonally wet forest at 2050 m altitude in the Kumaun Himalaya: Cedrus deodora, Quercus floribunda and Q. leucotrichophora. The total biomass and species number per unit trunk area, were found to increase with trunk size. It was supposed that the results indicated a succession in the type of epiphytic cover from young trunks to older trunks.The amount of loose material (plant remains and soil) per unit area of trunk increased with increasing girth. The C:N ratio in this material was initially very high on the oaks (129–197) and declined with increasing trunk size (to 73–78); the ratio was constant across girth classes in the cedar (86–87).Bryophytes produced most biomass on most trunks; next to them were lichens on the smallest trunks, and flowering plants on the largest. The number of species of epiphytes was similar on all three host species. The results are discussed in relation to contemporary ideas on diversity and strategies.Nomenclature follows: Ganguli (1972, 1977 & 1980); Kashyap (1929); Nayar (1970); Osmaston (1927).Financial support from the Council of Scientific and Industrial Research, Delhi and University Grants Commission, New Delhi is gratefully acknowledged.     

Right bundle branch block pattern after uncomplicated right ventricular outflow tract pacing in a patient with a left sided superior vena cava and corrected tetralogy of Fallot

Usually an electrocardiogram after right ventricular (RV) pacing should yield left bundle branch block (LBBB) pattern. However, the presence of right bundle branch block (RBBB) pattern after pacemaker implantation should alert the physician to a malposition of lead. We report a case of 18-year-old female who underwent dual chamber pacemaker implantation and had RBBB pattern post implantation. Detailed evaluation revealed an uncomplicated right ventricular outflow tract pacing. The possible causes of this abnormal pattern after an uncomplicated RV pacing are also reviewed.     

Urea‐induced binding between diclofenac sodium and bovine serum albumin: a spectroscopic insight

We investigated the interaction of diclofenac sodium (Dic.Na) with bovine serum albumin (BSA) in the absence and presence of urea using different spectroscopic techniques. A fluorescence quenching study revealed that the Stern–Volmer quenching constant decreases in the presence of urea, decreasing further at higher urea concentrations. The binding constant and number of binding sites were also evaluated for the BSA–Dic.Na interaction system in the absence and presence of urea using a modified Stern–Volmer equation. The binding constant is greater at high urea concentrations, as shown by the fluorescence results. In addition, for the BSA–Dic.Na interaction system, a static quenching mechanism was observed, which was further confirmed using time‐resolved fluorescence spectroscopy. UV–vis spectroscopy provided information about the formation of a complex between BSA and Dic.Na. Circular dichroism was carried out to explain the conformational changes in BSA induced by Dic.Na in the absence and presence of urea. The presence of urea reduced the α‐helical content of BSA as the Dic.Na concentration varied. The distance r between the donor (BSA) and acceptor (Dic.Na) was also obtained in the absence and presence of urea, using fluorescence resonance energy transfer. Copyright © 2015 John Wiley & Sons, Ltd.