Chemoreceptor sensitivity and maladaptation to high altitude in man

Studies were carried out to find out the role of chemoreceptor sensitivity in the causation of maladaptation syndromes on acute exposure to altitude. The experiments were done in two phases. In phase I, the responses in chemoreceptor sensitivity were studied in altitude acclimatized subjects and compared with those who suffered from either High Altitude Pulmonary Oedema (HAPO) or Acute Mountain Sickness (AMS). In Phase II, a similar comparison was done in two groups of subjects, one representing normal sojourners at 3,500 m and the other being subjects who had just recovered from HAPO. The first phase was done at Delhi; and the second at an altitude of 3,500 m. Parameters of assessment were hypoxic sensitivity, carbon dioxide sensitivity, ventilation (VE), respiratory frequency (Rf), forced vital capacity (FVC), forced expiratory volume at the first second (FEV1), heart rate (HR), blood pressure (BP), and oral temperature (Tor). The results showed significantly lower sensitivity to both hypoxia and carbon dioxide in maladapted subjects, as compared to those who were acclimatized in both the categories suggesting thereby that reduced chemoreceptor sensitivity might be an initiating factor in the causation of maladaptation syndromes at altitude.     

Purification and properties of valyl-tRNA synthetase from Mycobacterium smegmatis.

Valyl-tRNA synthetase from Mycobacterium smegmatis has been purified over 1200-fold by conventional techniques as well as affinity chromatography on valyl-aminohexyl Sepharose columns. The purified preparation is homogeneous by electrophoretic and immunologic criteria. The enzyme is a tetramer of approximate molecular weight of 120,000, composed of a single type of subunit. The synthetase exhibited maximal activity between 35--40 degrees C and pH 6.8--7.0. The pure enzyme though stable for several months below 0 degrees C, loses activity completely at 70 degrees C, for 1 min. The enzyme showed normal Michaelis-Menten kinetic behaviour in the total aminoacylation reaction with Km values of 1.25 microM, 0.1 mM and 1.0 microM for valine, ATP and tRNA, respectively, but the kinetic response deviated from the above pattern in the partial (activation) reaction. Based on these findings, the existence of the enzyme in two molecular forms, modulated by substrate concentration has been suggested; of these, only one may be active in the total reaction, while both forms may function in the phophosphate exchange reaction.     

SB203580 Modulates p38 MAPK Signaling and Dengue Virus-Induced Liver Injury by Reducing MAPKAPK2, HSP27, and ATF2 Phosphorylation

Dengue virus (DENV) infection causes organ injuries, and the liver is one of the most important sites of DENV infection, where viral replication generates a high viral load. The molecular mechanism of DENV-induced liver injury is still under investigation. The mitogen activated protein kinases (MAPKs), including p38 MAPK, have roles in the hepatic cell apoptosis induced by DENV. However, the in vivo role of p38 MAPK in DENV-induced liver injury is not fully understood. In this study, we investigated the role of SB203580, a p38 MAPK inhibitor, in a mouse model of DENV infection. Both the hematological parameters, leucopenia and thrombocytopenia, were improved by SB203580 treatment and liver transaminases and histopathology were also improved. We used a real-time PCR microarray to profile the expression of apoptosis-related genes. Tumor necrosis factor α, caspase 9, caspase 8, and caspase 3 proteins were significantly lower in the SB203580-treated DENV-infected mice than that in the infected control mice. Increased expressions of cytokines including TNF-α, IL-6 and IL-10, and chemokines including RANTES and IP-10 in DENV infection were reduced by SB203580 treatment. DENV infection induced the phosphorylation of p38MAPK, and its downstream signals including MAPKAPK2, HSP27 and ATF-2. SB203580 treatment did not decrease the phosphorylation of p38 MAPK, but it significantly reduced the phosphorylation of MAPKAPK2, HSP27, and ATF2. Therefore, SB203580 modulates the downstream signals to p38 MAPK and reduces DENV-induced liver injury.     

Enhanced glutamate,IP3 and cAMP activity in the cerebral cortex of Unilateral 6-hydroxydopamine induced Parkinson's rats: Effect of 5-HT,GABA and bone marrow cell supplementation

Parkinson's disease is characterized by progressive cell death in the substantia nigra pars compacta, which leads to dopamine depletion in the striatum and indirectly to cortical dysfunction. Increased glutamatergic transmission in the basal ganglia is implicated in the pathophysiology of Parkinson's disease and glutamate receptor mediated excitotoxicity has been suggested to be one of the possible causes of the neuronal degeneration. In the present study, the effects of serotonin, gamma-aminobutyric acid and bone marrow cells infused intranigrally to substantia nigra individually and in combination on unilateral 6-hydroxydopamine induced Parkinson's rat model was analyzed. Scatchard analysis of total glutamate and NMDA receptor binding parameters showed a significant increase in B_max (P < 0.001) in the cerebral cortex of 6-hydroxydopamine infused rat compared to control. Real Time PCR amplification of NMDA2B, mGluR5, bax, and ubiquitin carboxy-terminal hydrolase were up regulated in cerebral cortex of 6-hydroxydopamine infused rats compared to control. Gene expression studies of GLAST, ά-Synuclien and Cyclic AMP response element-binding protein showed a significant (P < 0.001) down regulation in 6-OHDA infused rats compared to control. Behavioural studies were carried out to confirm the biochemical and molecular studies. Serotonin and GABA along with bone marrow cells in combination showed reversal of glutamate receptors and behaviour abnormality shown in the Parkinson's rat model. The therapeutic significance in Parkinson's disease is of prominence.     

Purification and kinetic mechanism of lysyl-tRNA synthetase from Mycobacterium smegmatis SN2

Lysyl-tRNA synthetase [L-lys:tRNAlys ligase (AMP forming) EC:6.1.1.6] has been purified to homogeneity from Mycobacterium smegmatis SN2. The enzyme is a dimer of molecular weight 126,000 and is composed of identical subunits. A detailed analysis of the kinetic mechanism of the lysyl-tRNA synthetase has been carried out. A rapid equilibrium random ter ter mechanism is proposed based on initial velocity and product inhibition studies. There is no evidence for the formation of enzyme-bound lysyl-adenylate. The reverse reaction, studied by the deacylation of lysyl-tRNA, requires the presence of both AMP and PPi. This observation is consistent with the mechanism proposed.     

Transfection of Mycobacterium smegmatis SN2 with mycobacteriophage I3 DNA

Mycobacterium smegmatis SN2 does not exhibit natural competence for the uptake of phage I3 DNA. Competence can artificially be induced by treatment with glycine or CaCl₂, and the combination of both is even more effective. The efficiency of transfection can be improved by inclusion of protamine sulphate and heterologous RNA in the system. From ³²P DNA uptake studies the major barrier for the entry of DNA has been found to be the complex cell wall. The efficiency of transfection calculated on the basis of fraction of DNA which has entered the cell is comparable to that of other bacterial systems. The phage development takes a longer time (7 h for one cycle) after transfection, as compared to infection (4 h).