Controlled initiation of chromosomal replication in Escherichia coli requires functional Hda protein

Regulatory inactivation of DnaA helps ensure that the Escherichia coli chromosome is replicated only once per cell cycle, through accelerated hydrolysis of active replication initiator ATP-DnaA to inactive ADP-DnaA. Analysis of deltahda strains revealed that the regulatory inactivation of DnaA component Hda is necessary for maintaining controlled initiation but not for cell growth or viability.     

Physical Activity Energy Expenditure of Adolescents in India

Physical activity (PA) has rarely been quantified in adolescent populations undergoing economic transition; therefore relationships with disease still remain uncertain. This study assessed whether absolute PA energy expenditure (PAEE), PAEE/kg, and PAEE/kg_FFM could be accurately estimated using accelerometry and a questionnaire in Indian adolescents and how these values compared to those of other populations. PAEE was assessed using doubly labeled water (DLW) in 30 adolescents from Chennai, India, over seven consecutive days, simultaneous with the measurement of PA using accelerometry and a previous‐week recall questionnaire. Accelerometry counts (regression analysis) and questionnaire data were used to estimate PAEE; estimates were cross‐validated using the Bland‐Altman method. Accelerometry data and DLW‐derived PAEE were visually compared to values from four North American and European populations. For boys, 49% of the variance in DLW‐derived PAEE was explained with an equation including accelerometry counts and fat‐free mass (FFM). Questionnaire‐derived estimates did not contribute to the explained variance in DLW derived PAEE. The group‐level PA of these Indian adolescents was successfully assessed using accelerometry, but not questionnaire. DLW‐derived PAEE/kg_FFM (mean (s.d.): 53.0 (27.5) kJ/kg_FFM/day) was lower in this group than other adolescent populations in Europe and similar to those in North America. Additionally, four boys and none of the girls accumulated ≥60 min/day of accelerometry‐derived moderate intensity activity, indicating low levels of PAEE and PA in these adolescents. Further research is necessary to investigate the association between PA and health outcomes in Indian adolescents.     

Subcellular localization of ammonium transporters in <Emphasis Type="Italic">Dictyostelium discoideum</Emphasis>

Background  

With the exception of vertebrates, most organisms have plasma membrane associated ammonium transporters which primarily serve to import a source of nitrogen for nutritional purposes. Dictyostelium discoideum has three ammonium transporters, Amts A, B and C. Our present work used fluorescent fusion proteins to determine the cellular localization of the Amts and tested the hypothesis that the transporters mediate removal of ammonia generated endogenously from the elevated protein catabolism common to many protists.     

The Fis Protein Has a Stimulating Role in Initiation of Replication in Escherichia coli In Vivo

The Fis protein is a nucleoid associated protein that has previously been reported to act negatively in initiation of replication in Escherichia coli. In this work we have examined the influence of this protein on the initiation of replication under different growth conditions using flow cytometry. The Fis protein was found to be increasingly important with increasing growth rate. During multi-fork replication severe under-initiation occurred in cells lacking the Fis protein; the cells initiated at an elevated mass, had fewer origins per cell and the origins were not initiated in synchrony. These results suggest a positive role for the Fis protein in the initiation of replication.     

Diversity of Sulfur Isotope Fractionations by Sulfate-Reducing Prokaryotes

Batch culture experiments were performed with 32 different sulfate-reducing prokaryotes to explore the diversity in sulfur isotope fractionation during dissimilatory sulfate reduction by pure cultures. The selected strains reflect the phylogenetic and physiologic diversity of presently known sulfate reducers and cover a broad range of natural marine and freshwater habitats. Experimental conditions were designed to achieve optimum growth conditions with respect to electron donors, salinity, temperature, and pH. Under these optimized conditions, experimental fractionation factors ranged from 2.0 to 42.0‰. Salinity, incubation temperature, pH, and phylogeny had no systematic effect on the sulfur isotope fractionation. There was no correlation between isotope fractionation and sulfate reduction rate. The type of dissimilatory bisulfite reductase also had no effect on fractionation. Sulfate reducers that oxidized the carbon source completely to CO₂ showed greater fractionations than sulfate reducers that released acetate as the final product of carbon oxidation. Different metabolic pathways and variable regulation of sulfate transport across the cell membrane all potentially affect isotope fractionation. Previous models that explained fractionation only in terms of sulfate reduction rates appear to be oversimplified. The species-specific physiology of each sulfate reducer thus needs to be taken into account to understand the regulation of sulfur isotope fractionation during dissimilatory sulfate reduction.     

Linkage of type I interferon activity and TNF-alpha levels in serum with sarcoidosis manifestations and ancestry

Background

Both type I interferon (IFN), also known as IFN-α and tumor necrosis factor alpha (TNF-α) have been implicated in the pathogenesis of sarcoidosis. We investigated serum levels of these cytokines in a large multi-ancestral sarcoidosis population to determine correlations between cytokine levels and disease phenotypes.

Methods

We studied serum samples from 98 patients with sarcoidosis, including 71 patients of African-American ancestry and 27 patients of European-American ancestry. Serum type I IFN was measured using a sensitive reporter cell assay and serum TNF-α was measured using a commercial ELISA kit. Clinical data including presence or absence of neurologic, cardiac, and severe pulmonary manifestations of sarcoidosis were abstracted from medical records. Twenty age-matched non-autoimmune controls were also studied from each ancestral background. Differences in cytokine levels between groups were analyzed with Mann-Whitney U test, and correlations were assessed using Spearman''s rho. Multivariate logistic regression models were used to detect associations between cytokines and clinical manifestations.

Results

Significant differences in cytokine levels were observed between African- and European-American patients with sarcoidosis. In African-Americans, serum TNF-α levels were significantly higher relative to matched controls (P = 0.039), and patients with neurologic disease had significantly higher TNF-α than patients lacking this manifestation (P = 0.022). In European-Americans, serum type I IFN activity was higher in sarcoidosis cases as compared to matched controls, and patients with extra-pulmonary disease represented a high serum IFN subgroup (P = 0.0032). None of the associations observed were shared between the two ancestral groups.

Conclusions

Our data indicate that significant associations between serum levels of TNF-α and type I IFN and clinical manifestations exist in a sarcoidosis cohort that differ significantly by self-reported ancestry. In each ancestral background, the cytokine elevated in patients with sarcoidosis was also associated with a particular disease phenotype. These findings may relate to ancestral differences in the molecular pathogenesis of this heterogeneous disease.