Glutamine synthetase of pea leaves. I. Purification, stabilization, and pH optima

1. Glutamine synthetase [l-glutamate: ammonia ligase (ADP), E.C. 6.3.1.2]was purified to apparent homogeneity from the shoots of light-grown pea seedlings. It was found to be quite unstable, but. could be partially stabilized by the addition of divalent cation (Mg²⁺ or Mn²⁺), and still further by the addition of sucrose (0.5–1.5 m), fructose (1–2.5 m), or ethyleneglycol (20–30%). Stability varied considerably depending on pH and whether Mg²⁺ or Mn²⁺ was used. Under certain incubation conditions the inactivated enzyme could be partially reactivated. 2. The pH optimum for glutamine synthesis varied widely (5.0–8.2) depending on the type and concentration of divalent cation (Mn²⁺, Co²⁺, Mg²⁺) and the ATP concentration.     

Asymptomatic Significant Bacteriuria in the Non-pregnant Woman: II. Response to Treatment and Follow-up

Short courses of nitrofurantoin and ampicillin produced an immediate cure in 80% of adult non-pregnant bacteriuric women. Of the subjects so treated, 55% remained cured at the end of one year. Over the same follow-up period 36% of untreated bacteriuric women developed a spontaneous remission of bacteriuria. Treatment failed to prevent the development of symptomatic infection, and the reinfections which followed successful treatment were more commonly associated with the development of symptoms than the persistent or relapsing infections in untreated or unsuccessfully treated subjects.It is concluded that a search for bacteriuria in non-pregnant women is unlikely to be of value as a preventive measure, since in many instances it fails to detect urinary tract infection at an early stage and since treatment by methods suitable for large-scale use is ineffective.     

Effects of ovine LH, GH and prolactin, and testosterone on serum testosterone and estradiol-17 beta levels, and seminal vesicle and testicular activity in the catfish Clarias batrachus (L.)

Intraperitoneal administrations of testosterone (0.5 microgram/g body wt), and ovine LH (1.0 microgram/g body wt), GH (5 micrograms/g body wt) and prolactin (10 micrograms/g body wt) daily for 7 days during early prespawning phase (May) in C. batrachus produced varied effects on seminal vesicle (SVSI) and testicular (GSI) weights and biochemical correlates. Testosterone and LH treatments significantly increased serum testosterone level and concentrations of total proteins, fructose, hexosamines and sialic acid in both seminal vesicles and testis. Serum E2 levels increased significantly only after testosterone treatment. GH treatment increased significantly serum testosterone level and only the concentrations of SV hexosamines and testicular protein. Prolactin, however, significantly lowered serum testosterone level and concentrations of total protein, hexosamines in both SV and testis, and testicular fructose and sialic acid levels. The results show that the stimulating effect of LH and GH on SV and testicular activity is mediated through the increased secretion of testosterone and the inhibitory effect of prolactin by decreased testosterone secretion.     

Root dynamics of Carex stricta-dominated tussock meadows

Background and aims

The roots of tussock-forming plants contribute to the formation of microtopographic features in many ecosystems, but the dynamics of such roots are poorly understood. We examined the spatial heterogeneity of tussock fine root dynamics to investigate allocation patterns and the role of root productivity in the persistence of tussock structures.

Methods

We compared the spatial variability of fine root (<1 mm, 1–2 mm) density, biomass, % live, allocation, turnover rate (using bomb 14C), and productivity of four Carex stricta Lam.-dominated tussock meadows in the upper Midwest, USA (3 reference, 1 restored site).

Results

Relative to underlying microsites, tussocks were warm, dry, and high in root density, productivity, % live biomass, and turnover. Root productivity averaged 649 g?m^?2 yr^?1 (±208) in reference sites, comprised 57 % (±10) of total net production, and was concentrated in tussocks (70 %?±?4). Root turnover rate averaged 0.63 yr^?1 (±0.08), but tussocks had ~50 % faster root turnover than the underlying soil, and <1 mm roots turned over ~40 % faster than 1–2 mm roots.

Conclusions

Our detailed analysis of the spatial heterogeneity of tussock root dynamics suggests that high allocation and elevated turnover of tussock roots facilitates organic matter accumulation and tussock persistence over time.     

Therapeutic effect of ethanolic extract of Hygrophila spinosa T. Anders on gentamicin-induced nephrotoxicity in rats

Therapeutic effect of ethanolic extract of Hygrophila spinosa in gentamicin-induced nephrotoxic model of kidney injury in male Sprague-Dawley rats was studied. Rats were administered with gentamicin at a dose of 80 mg/kg intraperitoneally (ip) to induce nephrotoxicity. Kidney function was assessed by measuring serum creatinine and urea. Kidney superoxide dismutase, lipid peroxidation, catalase and reduced glutathione were also measured in control and treated rats. H. spinosa extract showed free radical scavenging activities at doses of 50 and 250 mg/kg with a predominant activity at 250 mg/kg. The ethanolic extract also caused a reduction in serum creatinine and urea levels. Histopathological studies were conducted to confirm the therapeutic action of the plant extract. The results demonstrated that the ethanolic extract of whole plant of H. spinosa evinced the therapeutic effect and inhibited gentamicin-induced proximal tubular necrosis.     

Risk Factors for MDR and XDR-TB in a Tertiary Referral Hospital in India

Background

India has a high burden of drug resistant TB, although there are few data on XDR-TB. Although XDR-TB has existed previously in India, the definition has not been widely applied, and surveillance using second line drug susceptibility testing has not been performed. Our objective was to analyze clinical and demographic risk factors associated with isolation of MDR and XDR TB as compared to susceptible controls, at a tertiary center.

Methodology/Findings

Retrospective chart review based on positive cultures isolated in a high volume mycobacteriology laboratory between 2002 and 2007. 47 XDR, 30 MDR and 117 susceptible controls were examined. Drug resistant cases were less likely to be extrapulmonary, and had received more previous treatment regimens. Significant risk factors for XDR-TB included residence outside the local state (OR 7.43, 3.07-18.0) and care costs subsidized (OR 0.23, 0.097-0.54) in bivariate analysis and previous use of a fluoroquinolone and injectable agent (other than streptomycin) (OR 7.00, 95% C.I. 1.14-43.03) and an initial treatment regimen which did not follow national guidelines (OR 5.68, 1.24-25.96) in multivariate analysis. Cavitation and HIV did not influence drug resistance.

Conclusions/Significance

There is significant selection bias in the sample available. Selection pressure from previous treatment and an inadequate initial regimen increases risk of drug resistance. Local patients and those requiring financial subsidies may be at lower risk of XDR-TB.     

Correlation between cell killing and massive second-round superinfection by members of some subgroups of avian leukosis virus.

Avian leukosis viruses of subgroups B, D, and F are cytopathic for chicken cells, whereas viruses of subgroups A, C, and E are not. The amounts of unintegrated linear viral DNA in cells at different times after infection with cytopathic or noncytopathic viruses were determined by hybridization and transfection assays. Shortly after infection, there is a transient accumulation of unintegrated linear viral DNA in cells infected with cytopathic avian leukosis viruses. By 10 days after infection, the majority of this unintegrated viral DNA is not present in the infected cells. The transient cytopathic effect seen in these infected cells also disappears by this time. Low amounts of unintegrated linear viral DNA persist in these cells. Cells infected with noncytopathic viruses do not show this transient accumulation of unintegrated viral DNA. Cells infected with cytopathic viruses and subsequently grown in the presence of neutralizing antibody do not show the transient accumulation of unintegrated viral DNA or cytopathic effects. These results demonstrate a correlation between envelope subgroup, transient accumulation of unintegrated linear viral DNA, and transient cell killing by avian leukosis viruses. The cell killing appears to be the result of massive second-round superinfection by the cytopathic avian leukosis viruses.     

Microbial community analysis of a coastal salt marsh affected by the Deepwater Horizon oil spill

Coastal salt marshes are highly sensitive wetland ecosystems that can sustain long-term impacts from anthropogenic events such as oil spills. In this study, we examined the microbial communities of a Gulf of Mexico coastal salt marsh during and after the influx of petroleum hydrocarbons following the Deepwater Horizon oil spill. Total hydrocarbon concentrations in salt marsh sediments were highest in June and July 2010 and decreased in September 2010. Coupled PhyloChip and GeoChip microarray analyses demonstrated that the microbial community structure and function of the extant salt marsh hydrocarbon-degrading microbial populations changed significantly during the study. The relative richness and abundance of phyla containing previously described hydrocarbon-degrading bacteria (Proteobacteria, Bacteroidetes, and Actinobacteria) increased in hydrocarbon-contaminated sediments and then decreased once hydrocarbons were below detection. Firmicutes, however, continued to increase in relative richness and abundance after hydrocarbon concentrations were below detection. Functional genes involved in hydrocarbon degradation were enriched in hydrocarbon-contaminated sediments then declined significantly (p<0.05) once hydrocarbon concentrations decreased. A greater decrease in hydrocarbon concentrations among marsh grass sediments compared to inlet sediments (lacking marsh grass) suggests that the marsh rhizosphere microbial communities could also be contributing to hydrocarbon degradation. The results of this study provide a comprehensive view of microbial community structural and functional dynamics within perturbed salt marsh ecosystems.     

Multiple transporters associated with malaria parasite responses to chloroquine and quinine

Mutations and/or overexpression of various transporters are known to confer drug resistance in a variety of organisms. In the malaria parasite Plasmodium falciparum, a homologue of P-glycoprotein, PfMDR1, has been implicated in responses to chloroquine (CQ), quinine (QN) and other drugs, and a putative transporter, PfCRT, was recently demonstrated to be the key molecule in CQ resistance. However, other unknown molecules are probably involved, as different parasite clones carrying the same pfcrt and pfmdr1 alleles show a wide range of quantitative responses to CQ and QN. Such molecules may contribute to increasing incidences of QN treatment failure, the molecular basis of which is not understood. To identify additional genes involved in parasite CQ and QN responses, we assayed the in vitro susceptibilities of 97 culture-adapted cloned isolates to CQ and QN and searched for single nucleotide polymorphisms (SNPs) in DNA encoding 49 putative transporters (total 113 kb) and in 39 housekeeping genes that acted as negative controls. SNPs in 11 of the putative transporter genes, including pfcrt and pfmdr1, showed significant associations with decreased sensitivity to CQ and/or QN in P. falciparum. Significant linkage disequilibria within and between these genes were also detected, suggesting interactions among the transporter genes. This study provides specific leads for better understanding of complex drug resistances in malaria parasites.