Purification and properties of mycobacterial GDP-mannose pyrophosphorylase

The enzyme that catalyzes the formation of GDP-d-mannose from GTP and alpha-d-mannose-1-P was purified about 2300-fold to near homogeneity from the soluble fraction of Mycobacterium smegmatis. At the final stage of purification, a major protein band of 37 kDa was observed and this band was specifically labeled, and in a concentration-dependent manner, by the photoaffinity probe 8-N3-GDP[32P]-d-mannose. The purified enzyme was stable for several months when kept in the frozen state. The 37-kDa band was subjected to protein sequencing and one peptide sequence of 25 amino acids showed over 80% identity to GDP-mannose pyrophosphorylases of pig liver and Saccharomyces cerevesiae. In contrast to some other bacterial GDP-mannose pyrophosphorylases, the mycobacterial enzyme was not multifunctional and did not have phosphomannose isomerase or phosphoglucose isomerase activity. Also, in contrast to the pig liver enzyme which uses mannose-1-P or glucose-1-P plus GTP to synthesize either GDP-mannose or GDP-glucose, the mycobacterial enzyme was specific for mannose-1-P as the sugar phosphate substrate. The enzyme was also relatively specific for GTP as the nucleoside triphosphate substrate. ITP was about 18% as effective as GTP, but ATP, CTP, and UTP were inactive. The activity of the enzyme was inhibited by GDP-glucose and glucose-1-P, although neither was a substrate for this enzyme. The pH optimum for the enzyme was 8.0, and Mg2+ was the best cation with optimum activity at about 5 mM. This enzyme is important for producing the activated form of mannose for formation of cell wall lipoarabinomannan and various mannose-containing glycolipids and polysaccharides.     

Self-incompatibility alleles from Physalis: implications for historical inference from balanced genetic polymorphisms

Balanced genetic polymorphism has been proposed as a source from which to infer population history complementary to that of neutral genetic polymorphism, because genetic polymorphism maintained by balancing selection permits inferences about population size over much longer spans of time. However, empirical data for both S genes and major histocompatibility complex genes do not fit expectations of coalescent theory. Species-specific gene genealogies have longer terminal branches than expected, indicating an apparent slowdown in the origination of new alleles. Here, we present evidence that divergent S alleles were selectively maintained in Physalis cinerascens during a reduction in population size, generating longer terminal branches in the S gene genealogy relative to the congener Physalis crassifolia. Retention of divergent alleles during reduction in the number of alleles violates assumptions of the coalescent model used to estimate effective population size. Recent theoretical and empirical results are consistent with the proposition that nonrandom sorting is a general property of balanced genetic polymorphisms, suggesting that studies of balanced polymorphism that infer the absence of population bottlenecks may overestimate effective population size.     

ON THE MOMENT COEFFICIENT CALCULATION OF A ROTATING DISK IN A TURBULENT FLUID FLOW

Mixing in co-rotating sinusoidal cavity flows is studied with period, T, and phase shift, a, as parameters. Notice that the main obstacle to uniform mixing is the existence of stable :ower periodic orbits, mixing windows, where uniform mixing takes place, are found in the T— a space u:iing bifurcation analysis. The main advantage of this method is a great saving of the computation time.     

Accessory artificial circulation in complex treatment of experimental acute toxic pulmonary edema

In experiments on 50 dogs with toxic acute edema of the lung, induced with intravenous injection of 0.1% silver nitrate, the authors have studied the efficacy of accessory artificial circulation and "conservative" therapy. During the perfusion a discharge of the right portions, adequate extracorporeal gas metabolism, normalization of blood gas and acid-base balance were noted; an intensity of pulmonary edema is descreased. An intensive therapy for pulmonary edema was found to be more effective in association of "conservative" treatment with venoarterial perfusion and blood oxygenation.     

Nitrous oxide "analgesia": resemblance to opiate action

Nitrous oxide produced a dose-related "analgesia" in mice (median effective dose, 55 percent). The analgesia was evaluated by means of a phenylquinone writhing test. Narcotic antagonists or chronic morphinization reduced nitrous oxide analgesia. Either nitrous oxide releases an endogenous analgesic or narcotic antagonists have analgesic antagonist properties heretofore unappreciated.     

Measurement and Simulation of the Flow Field around a Triangular Lattice Meteorological Mast

The international standard IEC 61400-12-1 Wind turbines--Part 12-1： Power performance measurements of electricity producing wind turbines＂ aims to provide a uniform methodology that will ensure consistency, accuracy and reproducibility in the measurement and analysis of power performance by wind turbines. Annex G of this standard provides a methodology for the appropriate arrangement of instruments on the meteorological mast to ensure accurate measurement. For cup anemometers it provides recommendations about their location relative to the mast so that the effect of mast and boom interference on their output may be minimised. These recommendations are given for both tubular masts and lattice masts. This paper compares the flow distortion predicted by the IEC standard and the results of a 3D CFD （computational fluid dynamics） simulation of a triangular lattice mast. Based on the results of wind tunnel and CFD simulation it was found that the flow distortion surrounding the lattice mast was overpredicted by the method suggested in appendix G oflEC61400-12-1. Using the CFD data it was possible to determine, for a range of flow directions and mast heights, the distance from the mast that anemometers would need to be in order to be outside the flow distortion field.     

Stimulation of the extracellular signal-regulated kinase 1/2 pathway by human beta-3 adrenergic receptor: new pharmacological profile and mechanism of activation

(-)Deprenyl is an irreversible inhibitor of monoamine oxidase B (MAO-B) frequently used as an adjunct therapy in the treatment of Parkinson's Disease. Recent evidence, however, has found that deprenyl's metabolites are associated with an antiapoptotic action within certain neuronal populations. Interestingly, deprenyl's antiapoptotic actions appear not to depend upon the inhibition of MAO-B. Due to a paucity of information surrounding (-)deprenyl's ability to spare neurons in vivo, a series of studies was conducted to further investigate this phenomenon within an apoptotic neuronal death model: kainic acid induced excitotoxicity. Results indicated that (-)deprenyl increased hippocampal neuronal survival compared to saline-matched controls following kainic acid insult. Furthermore, it was discovered that (-)deprenyl treatment could be stopped 14 days following CNS insult by kainate, with evidence of neuronal sparing still present by day 28. In open-field locomotor activity testing of kainate-treated animals, those given subsequent (-)deprenyl treatment showed habituation curves similar to control subjects, while saline-treated animals did not. Given deprenyl's antiapoptotic actions, it is proposed that (-)deprenyl may be beneficial in the treatment of a variety of neurodegenerative diseases where evidence of apoptosis exists, such as Parkinson's and Alzheimer's Disease, by slowing the disease process itself.     

Aurothioglucose inhibits murine thioredoxin reductase activity in vivo

Gold (I)-containing compounds, including aurothioglucose (ATG), are potent in vitro inhibitors of several selenocysteine-containing enzymes. Gold compounds have also been shown to potentiate the virulence of several viruses in mice, including coxsackievirus, implicated as a possible infectious agent in Keshan disease. One possible mechanism by which gold compounds may be increasing the virulence of viral infections in mice is by acting as a selenium antagonist in vivo and inducing oxidative stress. To investigate the possible role of gold compounds in inducing oxidative stress in mice, we assessed the ability of ATG administered in vivo to inhibit the activity of the selenocysteine-containing enzymes thioredoxin reductase (TR) and glutathione peroxidase (GPX1). Doses as low as 0. 025 mg ATG/g body weight caused significant and prolonged inhibition of TR activity in all tissues examined. No such inhibition of GPX1 activity was seen, indicating differential in vivo sensitivity of the enzymes to inhibition by ATG. In liver and heart, some recovery of TR activity was observed after a 7-d period, but no recovery was observed in pancreas or kidney. Because TR is involved in several important cellular redox functions, its inhibition most likely will affect multiple cellular processes. These results indicate that in vivo administration of ATG results in significant and long-lasting inhibition of TR activity. Such inhibition of TR could lead to increased levels of oxidative stress in vivo, thereby increasing the virulence of several viruses including the coxsackievirus.