Studies on the efficacy of a phytohormone producing phosphate solubilizingBacillus firmus in augmenting paddy yield in acid soils of Nagaland

Summary A super strain ofBacillus firmus (NCIM-2636) producing a phytohormone, indole-3-acetic acid, in addition to its high ability to solubilize insoluble inorganic phosphates were applied in acid soils of Nagaland, India. Rice (Oryza sativa L.) variety Jaya and IR-8 were grown in kharif season in two successive years 1980 and 1981. After proper manuring the soils received single super phosphate (S.S.P.) and Mussoorie Rock phosphate (R.P.) separately at different doses. Yield of crop in both the years increased significantly due to bacterial inoculation. Maximum grain yield was recorded in Jaya variety under S.S.P. and R.P. when treatments were at the dose of 43.75 and 17.5 kg P ha⁻¹ respectively while the same in IR-8 variety under S.S.P. and R.P. treatments were at the dose of 35 and 17.5 kg P ha⁻¹ respectively. Maximum straw yield was produced by Jaya variety when 35 and 43.75 kg P ha⁻¹ in the form of S.S.P. and R.P. respectively were applied. Highest straw yield of IR-8 variety was obtained after the application of 17.5 kg P ha⁻¹ (S.S.P. and R.P.) in combination with phosphate solubilizing bacteria. Bacterial inoculation decreased the phosphorus availability in 1 st year but increased the same in 2nd year. Phosphorus content in grains was significantly enhanced in both the trials. Maximum uptake of phosphorus by grains was noted in Jaya variety at the dose of 47.5 kg P ha⁻¹ and in IR-8 variety at the dose of 52.5 kg P ha⁻¹ under S.S.P. treatment, while 8.75 and 35 kg P ha⁻¹ in the form of R.P. yielded similar results in Jaya and IR-8 varieties respectively. Phosphorus at the dose of 35 kg ha⁻¹ was found to cause more P-uptake by straw in both S.S.P. and R.P. treatments. The various data from the experiment conclusively proved that the bacterium in combination with R.P. produced the desired effect more prominently than when bacterium applied in combination with S.S.P.     

A continuous fluorimetric assay for ATPase activity.

A new continuous coupled fluorimetric assay is described for ATPases in general. Thus phosphate released from ATP hydrolysis is coupled to the nucleoside phosphorylase reaction using 7-methylguanosine as a fluorescent substrate for the nucleoside phosphorylase reaction. The hydrolysis of 7-methylguanosine leads to 7-methylguanine, which has lower quantum yield and hence can be used to monitor ATP hydrolysis continuously. The method has the potential to be extended to GTPase and nucleotidyltransferase assays.     

The Effects of Calpain Inhibition on IkBα Degradation After Activation of PBMCs: Identification of the Calpain Cleavage Sites

Human peripheral blood mononuclear cells (PBMCs) were activated using anti-CD3/CD28 (HIT3A/CD28.2) resulting in degradation of IkB alpha, an inhibitor of NFkB, relative to unactivated cells. Degradation of IkB alpha began by 30 min and proceeded for at least 5 h. Calpeptin, a calpain inhibitor, inhibited IkB alpha degradation in a time- and dose-dependent manner. Furthermore, calpain inhibition increased IkB alpha levels compared to nonactivated controls. Recombinant IkB alpha was incubated with purified porcine m-calpain in the presence of 0.1% Triton X-100, and the degradation products were monitored by SDS-PAGE and sequenced. Most of the degradation products were peptides derived from calpain, but one was derived from IkB alpha cleaved between amino acids 50 and 51 (glutamine and glutamic acid). The liberated fragment included the entire signal response domain (SRD), a region containing key serine and threonine residues necessary for phosphorylation by the IKKinase complex and sites required for ubiquitination. The results suggest that calpain plays an important role in IkB alpha degradation, a crucial event in T cell activation.     

PAHs,OCPs and PCBs in sediments from three catchments in Durban,South Africa

Polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) were analysed from sediment samples collected in 2012 from rivers, surface runoff canals and estuaries in three highly urbanised catchments in durban, KwaZulu-Natal, South Africa. PAHs were ubiquitous, at concentrations between 36–6 800 ng g^?1 dry mass (dm). Congener ratio diagnosis suggests the PAHs were derived predominantly from pyrogenic sources. Four OCPs and/or their metabolites were detected at varying frequencies and concentrations. Of these, dichlorodiphenyltrichloroethane (ddT) and metabolites were most frequently detected and were at a high concentration in sediment in some systems. Toxaphene was detected at a high concentration at some sites. The total PCB concentration varied widely, from below the method detection limit to 110 ng g^?1 (dm). Based on the comparison of chemical concentrations to international sediment quality guidelines, PAH, OCP and PCB concentrations in most sediment samples posed a low risk to sediment-dwelling organisms. However, in some instances the risk was moderate or high. It is recommended that these compounds be monitored more frequently and comprehensively in aquatic ecosystems to better understand the ecological and human health implications.     

Role of calpain in spinal cord injury: Increased calpain immunoreactivity in rat spinal cord after impact trauma

Impact spinal cord injury (20 g-cm) was induced in rat by weight drop. The immunoreactivity of mcalpain was examined in the lesion and adjacent areas of the cord following trauma. Increased calpain immunoreactivity was evident in the lesion compared to control and the immunostaining intensity progressively increased after injury. The calpain immunoreactivity was also increased in tissue adjacent to the lesion. mCalpain immunoreactivity was significantly stronger in glial and endothelial cells, motor neurons and nerve fibers in the lesion. The calpain immunoreactivity also increased in astrocytes and microglial cells in the adjacent areas. Proliferation of microglia and astrocytes identified by GSA histochemical staining and GFAP immunostaining, respectively, was seen at one and three days after injury. Many motor neurons in the ventral horn showed increased calpain immunoreactivity and were shrunken in the lesion. These studies indicate a pivotal role for calpain and the involvement of glial cells in the tissue destruction in spinal cord injury. Special issue dedicated to Dr. Marion E. Smith.     

Comparison of the Effect of Calpain Inhibitors on Two Extralysosomal Proteinases: The Multicatalytic Proteinase Complex and m-Calpain

Abstract: The potencies of three peptide aldehyde inhibitors of calpain (calpain inhibitors 1 and 2 and calpeptin) as inhibitors of four catalytic activities of the multicatalytic proteinase complex (MPC) were compared with their potencies as inhibitors of m-calpain. The chymotrypsinlike activity (cleavage after hydrophobic amino acids) and the caseinolytic activity (degradation of β-casein) of MPC were strongly inhibited by calpain inhibitors 1 and 2 (IC₅₀ values in the low micromolar range). Cleavage by MPC after acidic amino acids (peptidylglutamyl-peptide bond hydrolyzing activity) and basic amino acids (trypsinlike activity) was inhibited less effectively, declining moderately with increasing concentrations of calpain inhibitors 1 and 2. Calpeptin only weakly inhibited the four MPC activities, yet was the most potent inhibitor of m-calpain. These results indicate that caution must be exercised when calpain inhibitors 1 and 2 are used to infer calpain function. Calpeptin may be a better choice for such studies, although its effect on other cysteine or serine proteinases remains to be determined.     

The regional and subcellular distribution of calcium activated neutral proteinase (CANP) in the bovine central nervous system

Calcium-activated neutral proteinase (CANP) activity was determined in subcellular fractions and in different regions of bovine brain. The CANP specific activity in spinal cord and corpus callosum, areas rich in myelin, were almost six-fold greater than cerebral cortex and cerebellum. Treatment of whole homogenate and myelin with 0.1% Triton X-100 increased the CANP activity by tenfold. Subcellular fractions were prepared from bovine brain gray and white matter. Most of the CANP activity (70%) was in the primary particulate fractions P₁ (nuclear), P₂ (mitochondrial) and P₃ (microsomal). On subfractionation of each particulate fraction, the majority of the activity (greater than 50%) was recovered in the myelin-enriched fractions (P₁A, P₂A, P₃A) which separate at the interphase of 0.32 M- and 0l85 M-sucrose. The distribution of activity was P₂A>P₁A>P₃A. Further purification of myelin (of P₂A) increased the specific activity over homogenate by more than three-fold. The same myelin fractions contained the highest proportion (60%) and specific activity (five-fold increase) of CNPase. The enzyme activity in different regions of brain and in subcellular fractions was increased by 20–39% after the inhibitor was removed. Electron microscopic study confirmed that the myelin fractions were highly purified. The cytosolic fraction contained 20–30% of the total homogenate CANP activity. Other fractions contained low enzyme activity. CANP was identified in the purified myelin fraction by electroimmublot-technique. It is concluded that the bulk of CANP in CNS is tightly bound to the membrane, may be masked or hidden and is intimately associated with the myelin sheath.Abbreviations Used CANP calcium-activated neutral proteinase - CNPase adenosine-2, 3-cyclic nucleotide 3-phosphohydrolase     

Deciphering Parameter Sensitivity in the BvgAS Signal Transduction

To understand the switching of different phenotypic phases of Bordetella pertussis, we propose an optimized mathematical framework for signal transduction through BvgAS two-component system. The response of the network output to the sensory input has been demonstrated in steady state. An analysis in terms of local sensitivity amplification characterizes the nature of the molecular switch. The sensitivity analysis of the model parameters within the framework of various correlation coefficients helps to decipher the contribution of the modular structure in signal propagation. Once classified, the model parameters are tuned to generate the behavior of some novel strains using simulated annealing, a stochastic optimization technique.     

Degradation of cytoskeletal proteins in experimental spinal cord injury

Spinal cord injury was produced in rats by dropping a 10 g weight from 30 cm upon dura-invested exposed spinal cord. Examination of the fine structure of the traumatic lesion (15 min to 30 min) revealed granular degeneration of axons and occasional loosening of myelin lamellae. Older lesions (4 to 72 hours) showed degeneration of axons and vesiculation of myelin. At 15 minutes there is more loss of neurofilament proteins than of myelin proteins. Substantial decreases in the neurofilament and myelin proteins were observed at 30 minutes and the losses were even greater 2–72 hours after injury. This indicates that degeneration of axons may precede degradation of the myelin sheath and also that increased proteinase(s) activity, possibly activated by calcium, mediates the traumatic axonolysis and myelinolysis in experimental spinal cord trauma.