In vitro plant regeneration from cotyledonary nodes of Withania somnifera (L.) Dunal and assessment of clonal fidelity using RAPD and ISSR markers

An efficient large-scale clonal propagation protocol has been described for Withania somnifera (L.) Dunal, a valuable medicinal plant, using cotyledonary nodes derived from axenic seedlings. Murashige and Skoog’s (Physiol Plant 15:473–497, 1962) (MS) medium supplemented with 1.0 mg l^?1 N ⁶-benzyladenine (BA) was found to be optimum for production of multiple shoots (100 % shoot proliferation frequency and 16.93 shoots per explant). Successive shoot cultures were established by repeatedly sub-culturing the original cotyledonary node on a fresh medium after each harvest of newly formed shoots. Multiple shoot proliferation was also achieved from nodal segments derived from in vitro raised shoots on MS medium augmented with 1.0 mg l^?1 BA. Regenerated shoots were best rooted (95.2 %, 38.7 roots per shoot) in half-strength MS medium supplemented with 1.0 mg l^?1 indole-3-butyric acid. The plantlets were successfully acclimated and established in soil. Random amplified polymorphic DNA and inter-simple sequence repeats analysis revealed a homogeneous amplification profile for all micropropagated plants analyzed validating the genetic fidelity of the in vitro regenerated plants.     

Setaria digitata infections in cattle: parasite load, microfilaraemia status and relationship to immune response

A total of 110 cattle were examined in an area endemic for Bancroftian filariasis for the prevalence of infection of the bovine filarial parasite Setaria digitata. About 12.5% of cattle were found to harbour both adult worms in the peritoneum and microfilariae (mf) in circulation; 70% of the cattle were amicrofilaraemic but with an adult worm infection. A third group of cattle (16.5%) was free of detectable mf and adult worms. The presence of adult worms and/or mf did not influence the antibody levels to any of the four antigen preparations of S. digitata. However, there was a significant inverse relationship between the presence of antibodies to microfilarial sheaths and the absence of circulating mf as shown by the immunoperoxidase assay. Cattle immunoglobulin containing high titres of anti-sheath antibodies cleared circulating microfilariae very effectively in Mastomys coucha thus demonstrating the protective nature of anti-sheath antibodies in eliminating circulating microfilariae in vivo.     

Functional Interaction between the Scaffold Protein Kidins220/ARMS and Neuronal Voltage-Gated Na+ Channels

Kidins220 (kinase D-interacting substrate of 220 kDa)/ankyrin repeat-rich membrane spanning (ARMS) acts as a signaling platform at the plasma membrane and is implicated in a multitude of neuronal functions, including the control of neuronal activity. Here, we used the Kidins220^−/− mouse model to study the effects of Kidins220 ablation on neuronal excitability. Multielectrode array recordings showed reduced evoked spiking activity in Kidins220^−/− hippocampal networks, which was compatible with the increased excitability of GABAergic neurons determined by current-clamp recordings. Spike waveform analysis further indicated an increased sodium conductance in this neuronal subpopulation. Kidins220 association with brain voltage-gated sodium channels was shown by co-immunoprecipitation experiments and Na⁺ current recordings in transfected HEK293 cells, which revealed dramatic alterations of kinetics and voltage dependence. Finally, an in silico interneuronal model incorporating the Kidins220-induced Na⁺ current alterations reproduced the firing phenotype observed in Kidins220^−/− neurons. These results identify Kidins220 as a novel modulator of Na_v channel activity, broadening our understanding of the molecular mechanisms regulating network excitability.     

ZnAl2O4:Eu novel phosphor: SEM and mechanoluminescence characterization synthesized by solution combustion technique

Eu doped ZnAl₂O₄ phosphors were synthesized by the solution combustion technique using carbohydrazide as a fuel. Mechanoluminescence (ML) was excited impulsively by dropping a piston of 0.7 kg onto the phosphors. Two distinct peaks were observed in the ML glow curve of the γ‐ray irradiated ZnAl₂O₄:Eu phosphors. Dependence of ML on various parameters as impact velocity of the piston dropped on to it, mass of the sample, gamma ray doses given to the sample and ML spectra have been studied. ML emission spectrum showed the characteristic emission of Eu³⁺ ion in this system. ML is observed to be optimum for the sample having 0.2 mol% of Eu in the ZnAl₂O₄ phosphor. XRD result confirms formation of the phosphors. SEM characterization shows its surface morphology. This novel phosphor may be a potential candidate for dosimetric use due to its linear dose response. Copyright © 2014 John Wiley & Sons, Ltd.     

Galantamine alleviates inflammation and other obesity-associated complications in high-fat diet-fed mice

Obesity, a serious and growing health threat, is associated with low-grade inflammation that plays a role in mediating its adverse consequences. Previously, we have discovered a role for neural cholinergic signaling in controlling inflammation, and demonstrated that the cholinergic agent galantamine suppresses excessive proinflammatory cytokine release. The main objective of this study was to examine the efficacy of galantamine, a clinically-approved drug, in alleviating obesity-related inflammation and associated complications. After 8 wks on a high-fat diet, C57BL/6J mice were treated with either galantamine (4 mg/kg, intraperitoneally [i.p.]) or saline for 4 wks in parallel with mice on a low-fat diet and treated with saline. Galantamine treatment of obese mice significantly reduced body weight, food intake, abdominal adiposity, plasma cytokine and adipokine levels, and significantly improved blood glucose, insulin resistance and hepatic steatosis. In addition, galantamine alleviated impaired insulin sensitivity and glucose intolerance significantly. These results indicate a previously unrecognized potential of galantamine in alleviating obesity, inflammation and other obesity-related complications in mice. These findings are of interest for studying the efficacy of this clinically-approved drug in the context of human obesity and metabolic syndrome.     

Coupling dTTP hydrolysis with DNA unwinding by the DNA helicase of bacteriophage T7

The DNA helicase encoded by gene 4 of bacteriophage T7 assembles on single-stranded DNA as a hexamer of six identical subunits with the DNA passing through the center of the toroid. The helicase couples the hydrolysis of dTTP to unidirectional translocation on single-stranded DNA and the unwinding of duplex DNA. Phe(523), positioned in a β-hairpin loop at the subunit interface, plays a key role in coupling the hydrolysis of dTTP to DNA unwinding. Replacement of Phe(523) with alanine or valine abolishes the ability of the helicase to unwind DNA or allow T7 polymerase to mediate strand-displacement synthesis on duplex DNA. In vivo complementation studies reveal a requirement for a hydrophobic residue with long side chains at this position. In a crystal structure of T7 helicase, when a nucleotide is bound at a subunit interface, Phe(523) is buried within the interface. However, in the unbound state, it is more exposed on the outer surface of the helicase. This structural difference suggests that the β-hairpin bearing the Phe(523) may undergo a conformational change during nucleotide hydrolysis. We postulate that upon hydrolysis of dTTP, Phe(523) moves from within the subunit interface to a more exposed position where it contacts the displaced complementary strand and facilitates unwinding.     

Antigen adsorbed calcium phosphate nanoparticles stimulate both innate and adaptive immune response in fish, Labeo rohita H

Calcium phosphate nanoparticles as an antigen/protein delivery was explored in a fish model Labeo rohita H. S-layer protein (of Aeromonas hydrophila) adsorbed on nano sized calcium phosphate particles elicited both innate and adaptive immune parameters which persisted up to 63 days of post immunization through parenteral immunization and gave cross protections.     

Selection on GGU and CGU Codons in the High Expression Genes in Bacteria

The fourfold degenerate site (FDS) in coding sequences is important for studying the effect of any selection pressure on codon usage bias (CUB) because nucleotide substitution per se is not under any such pressure at the site due to the unaltered amino acid sequence in a protein. We estimated the frequency variation of nucleotides at the FDS across the eight family boxes (FBs) defined as Um(g), the unevenness measure of a gene g. The study was made in 545 species of bacteria. In many bacteria, the Um(g) correlated strongly with Nc′—a measure of the CUB. Analysis of the strongly correlated bacteria revealed that the U-ending codons (GGU, CGU) were preferred to the G-ending codons (GGG, CGG) in Gly and Arg FBs even in the genomes with G+C % higher than 65.0. Further evidence suggested that these codons can be used as a good indicator of selection pressure on CUB in genomes with higher G+C %.