Phenotypic and proteomic analysis of positively regulated gellan biosynthesis pathway in Sphingomonas elodea

Sphingomonas elodea is a Gram-negative bacterium capable of producing ‘gellan gum’ exopolysaccharide that is the most extensively studied expolysaccharides of microbial origin. In this study, we investigated the phenotypic and proteomic alterations in S. elodea by homogeneously expressing both gelA and gelN involved in positive regulation and extracellular secretion of metabolites in gellan biosynthesis, respectively. Expression of six histidine-tagged GelA and GelN was determined by Western blot analysis. Successful expression of GelA and GelN resulted in both morphological changes of colonies and enhanced secretion of gellan into the growth medium (GelA, 21.2% more and GelN, 48.3% more) overexpressed compared to the wile-type. Comparative two-dimensional gel electrophoresis analysis revealed a differential proteome expression in S. elodea overexpressing GelA and GelN. Proteins up- or down-regulated by GelA and GelN overexpression were found to be mainly sugar transportation proteins, two-component regulatory proteins, and proteins involved in secretion pathways. The results suggest that the effect of GelA and GelN overexpression on gellan biosynthesis might be mainly caused by increased transportation of sugar units or enhanced exportation of gellan.     

Parasitoids ofPectinophora gossypiella [Lep.: Gelechiidae] andEarias spp. [Lep.: Noctuidae] in the Punjab

Observations on the parasitoids of cotton bollworms in the Punjab were made during 1978 and 1979. The 2 trichogrammatid egg parasitoids, viz.Trichogramma achaeae Nagaraja & Nagarkatti andTrichogrammatoidea sp. nearguamensis Nagaraja (MS) were recovered fromPectinophora gossypiella (Saunders) in addition to 1 braconid egg-larval parasitoidChelonus sp. and 7 larval parasitoids, viz. 4 braconidsApanteles angaleti Muesebeck,Bracon greeni Ashsmead,Camptothlipsis sp. andRogas sp., 1 elasmid,Elasmus johnstoni Ferrière, 1 bethylid,Goniozus sp. and 1 ichneumonid,Scambus lineipes (Morley). FromEarias insulana Boisduval andEarias vittella F., 3 trichogrammatid egg parasitoids,T. achaeae, Trichogramma chilonis Ishii, andT. sp. nearguamensis 2 braconid larval parasitoids,B. greeni andRogas sp. and 1 chalcid pupal parasitoid,Brachymeria nephantidis Gahan, were recovered. One eurytomid hyperparasitoid,Eurytoma braconidis Ferrière was also recovered from the cocoons ofB. greeni. Of these parasitoids,T. achaeae, T. sp. nearguamensis, Camptothlipsis sp. andS. lineipes fromP. gossypiella, T. achaeae andB. nephantidis fromEarias spp. andE. braconidis fromB. greeni are new records.     

Pulmonary blood volume and edema in postpneumonectomy lung growth in rats

After pneumonectomy in young animals, the contralateral lung undergoes compensatory growth and generally attains the same weight and air space volume as both lungs in age-matched controls. In this study, we determined the contribution of lung edema and increased blood volume to the weight gain in rats. Three weeks after pneumonectomy (n = 18) or sham pneumonectomy (n = 17), the pulmonary blood volume and the extravascular water and albumin were evaluated by use of 51Cr-labeled erythrocytes and 125I-labeled albumin. The air space volume, blood-free lung weights, and DNA and protein content were also compared. The data show that the total pulmonary blood volumes and the blood volume per gram of blood-free dry lung were similar in pneumonectomized and age-matched sham controls. The total extravascular albumin and the extravascular albumin per gram of blood-free dry lung were also similar as well as the extravascular lung water, wet-to-dry weight ratios, DNA and protein content, and air space volumes. These data indicate that the increased weight of the postpneumonectomy lung was due to cellular and stromal proliferation. The blood volume and interstitial fluid increased in proportion to the increase in lung parenchyma. Neither vascular congestion nor increased extravascular protein and water contributed to the observed weight gain.     

Effects of velocity jumps on blood flow in large arteries

When a human being experiences a sudden velocity change, the blood flow is disturbed. A theoretical analysis to predict the effects of sudden velocity changes on blood flow in large arteries is presented. The situations is modelled as a one-dimensional flow problem in a viscoelastic tube where the fluid viscosity convective term in the equation of motion and nonlinearity in the elastic modulus of the tube wall are neglected. The governing equations of the model are solved by Laplace transformation. The computed results show that relatively high blood pressures, capable of harming circulation, are produced even by relatively moderate velocity jumps.     

Evaluation of the Pro-Lab ID ring system for the identification of medically important yeasts

We evaluated 151 coded isolates of medically important yeast species belonging to the genera Candida, Cryptococcus, Geotrichum, Rhodoturula, Saccharomyces and Torulopsis using the newly developed rapid Pro-Lab Identification Ring, PL 960 system (PLID-Ring). All isolates were concurrently identified by the API 20C and conventional procedures comprising macro- and micromorphology, assimilation and fermentation of various carbon and nitrogen compounds. The PLID-Ring system identified isolates of Candida albicans, C. kefyr, C. krusei, C. lusitaniae, C. parapsilosis, Rhodotorula rubra, and Torulopsis glabrata with 100% accuracy in 24 h. This system identified C guilliermondii and S. cerevisiae isolates with an accuracy of 90% and 86%, respectively, while those belonging to Cr. neoformans, T. candida (= C. famata), C. rugosa and C. tropicalis were identified with 38.4%, 50%, 12.5% and 50% accuracy, respectively. Three isolates of Cr. laurentii were not identified by the PLID-Ring system. The overall accuracy of the PLID-Ring system was 81.45% (123 of 151 isolates). However, the system does not include species such as Cr. laurentii in its data base. When these three Cr. laurentii isolates were excluded from the evaluation, the accuracy of the PLID-Ring system increased from 81.45% to 83.1%.     

Some measures of reducing leaching loss of nitrates beyond potential rooting zone

Summary Distribution patterns of nitrate in field are studied in twelve treatments comprising of different N splits and irrigation schedules, after the harvest of wheat. Total amount of irrigation and nitrogen application were kept same for each treatment. The curves show that heavy irrigation at greater intervals can result in larger amount of unutilised NO₃ ^–-N, which will eventually be lost beyond potential rooting zone. As irrigation becomes lighter and frequent, nitrates travel slowly and thus remain for more time within the reach of roots and are lost to a less extent. When whole of the nitrogen is applied in one lot, considerably more NO₃ ^–-N is lost under all the irrigation schedules. As the number of splits are increased, susceptibility of nitrate nitrogen for leaching decreases to a greater extent under lighter and more frequent irrigation schedule than the other. Besides N-splitting and irrigation criteria, efficiency and depth of rooting system of plants seems to play a major role in defining nitrate leaching patterns towards unsaturated zone.     

The structure of postsynaptic densities isolated from dog cerebral cortex: I. overall morphology and protein composition

A postsynaptic density (PSD) fraction, including some adherent subsynaptic web material, has been isolated from dog cerebral cortex by a short-procedure modification of methods of Davis and Bloom (21, 22) and Cotman and Taylor (20), using Triton X-100. The fraction has been visualized by thin-section, replica, and negative (phosphotungstic acid) staining electron microscopy and its proteins separated by high-resoltuion SDS gel electrophoresis. Morphologically, the preparation seems to be quite pure, with very little membrane contamination. The density is composed of protein, no nuclei acids, and very little phospholipids being detectable. The fraction had no ATPase or GTPase activity, but it did have a very small amount of cytochrome c oxidase activity (of a specific activity less than 0.5 percent that of a mitochondrial fraction) and a small amount of 5'- nucleotidase activity (of a specific activity between 6 and 7 percent that of a synaptic membrane fraction). Electron micrographs reveal cup-shaped structures approximately 400nm long and approximately 40nm wide, made up of apparent particles 13-28nm in diameter. However, en face views, and particularly micrographs of replicas and PTA-stained preparations, reveal a disk-shaped structure, outside diameter approximately 400 nm, in which filaments are seen to extend from the central part of the density. High resolution gel electrophoresis studies indicated some 15 major proteins and perhaps 10 or more minor ones; the predominant protein had a mol wt of 51,000, followed by ones at 45,000, 40,000, 31,000, 26,000, and several at 100,000. A comparison by gel electrophoresis of density fraction proteins with those of a lysed synaptosomal membrane fraction containing some adherent densities indicated some comigrating proteins, but the major membrane fraction protein, mol wt 52,000, was not found in the density fraction. Antibodies raised against the density fraction reacted with a preparation of solubilized synaptic membrane proteins. By both these criteria, it was considered that the density and the synaptic membrane have some proteins in common. By separately mixing (125)I-labeled myelin, synaptic vesicle, and mitochondrial fraction proteins with synaptosomes, and then isolating the density fraction from the mixture, it was concluded that a major 26,000 mol wt density fraction protein was common to both mitochondria and density, that none of the proteins of the density were contaminants from the mitochondrial fraction, that a minor approximately 150,000 band was a contaminant from the synaptic vesicle fraction, and that the moderately staining PSD fraction protein of 17,000 mol wt band was the result of contamination by the major basic protein of myelin. On the basis of the marker enzymatic assays and the mixing experiments, it is considered that the density fraction is moderately pure biochemically, and that its protein composition, aside from a few exceptions noted above, reflects its in situ character.