Effect of Obesity on Acute Ozone-Induced Changes in Airway Function, Reactivity, and Inflammation in Adult Females

We previously observed greater ozone-induced lung function decrements in obese than non-obese women. Animal models suggest that obesity enhances ozone-induced airway reactivity and inflammation. In a controlled exposure study, we compared the acute effect of randomized 0.4ppm ozone and air exposures (2 h with intermittent light exercise) in obese (N = 20) (30<BMI<40Kg/m2) vs. non-obese (N = 20) (BMI<25Kg/m2) non-smoking 18–35 year old women. We measured spirometry and bronchial reactivity to inhaled methacholine (3h post-exposure). Inflammation and obesity markers were assessed in the blood (pre, 4h post, and 20h post exposures) and induced-sputum (4h post-exposures and on 24h pre-exposure training day, no exercise): measures of C reactive protein (CRP) (blood only), leptin (blood only), adiponectin, interleukins IL-6, IL-1b, and IL-8, and tumor necrosis factor alpha, and sputum cell differential cell counts. The pre- to post-exposure decrease in forced vital capacity after ozone (adjusted for the change after air exposure) was significantly greater in the obese group (12.5+/-7.5 vs. 8.0+/-5.8%, p<0.05). Post ozone exposure, 6 obese and 6 non-obese subjects responded to methacholine at ≤ 10mg/ml (the maximum dose); the degree of hyperresponsiveness was similar for the two groups. Both BMI groups showed similar and significant ozone-induced increases in sputum neutrophils. Plasma IL-6 was increased by exercise (4 hr post air exposure vs. pre) only in the obese but returned to pre-air exposure levels at 20hr post-exposure. Plasma IL-6 was significantly increased at 4hr post ozone exposure in both groups and returned to pre-exposure levels by 20h post-exposure. These results confirm our previous findings of greater post-ozone spirometric decrements in obese young women. However, acute ozone-induced airway reactivity to methacholine and airway inflammation did not differ by obesity at the exposure and exercise levels used

Impact of recent cyclone lashed along West Bengal, Orissa and northern Andhra Pradesh coasts on the marine fishery sector

Impact of recent cyclone lashed along West Bengal, Orissa and northern Andhra Pradesh coasts on the marine fishery secto

Major and minor fisheries harbours of India. 8. Fisherries harbours along the coasts of North Andhra Pradesh, Orissa and West Bengal

The present account gives information about the three major and five minor fisheries harbours along the coasts of north Andhra Pradesh, Orissa and West Benga

Scientific, common and vernacular names of commercially important fin and shell fishes. 5. Orissa

Present paper focus on the scientific, common and vernacular names of important fin and shell fishes of Oriss

Not Available

Not AvailablePresent paper focus on the scientific, common and vernacular names of important fin and shell fishes of OrissaNot Availabl

Phosphorylation at Serines 216 and 221 Is Important for <i>Drosophila HeT-A</i> Gag Protein Stability

<div><p>Telomeres from <i>Drosophila</i> appear to be very different from those of other organisms – in size and the mechanism of their maintenance. In the absence of the enzyme telomerase, <i>Drosophila</i> telomeres are maintained by retrotransposition of three elements, <i>HeT-A</i>, <i>TART</i>, and <i>TAHRE</i>, but details of their transposition mechanisms are not known. Here we characterized some biochemical characteristics of the <i>HeT-A</i> Gag protein encoded by the <i>HeT-A</i> element to understand this mechanism. The <i>HeT-A</i> Gag protein when overexpressed in S2 cells was localized to the nucleus but was resistant to high salt, detergents and nuclease extraction treatments. Analysis of the <i>HeT-A</i> Gag protein by tandem mass spectrophotometry revealed that serines 216 and 221 are phosphorylated. Substituting these serines with alanine or aspartic acid by site-directed mutagenesis did not result in any changes in <i>HeT-A</i> Gag translocation across the nucleus, suggesting that phosphorylation of these sites is not associated with <i>HeT-A</i> Gag translocation, but time course experiments showed that these phosphorylation sites are important for Gag-protein stability.</p></div

Mutagenesis sites, primers and templates for site-directed mutagenesis.

<p>Mutagenesis sites, primers and templates for site-directed mutagenesis.</p

Tandem MS analysis of <i>HeT-A</i> Gag protein by mass spectrophometry.

<p>(A) MS/MS of ion m/z shows an extensive y-ion series and the presence of phosphorylation at serines 216 or 221, as indicated by the arrow. (B) Line diagram of different forms of <i>HeT-A</i> Gags (M1, M2, and M3) that were generated by site directed mutagenesis. Primer sets and template used for site directed mutagenesis are described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075381#pone-0075381-t002" target="_blank">Table 2</a>.</p

Time course of <i>HeT-A</i> Gag protein expression and proteasome treatment.

<p>(A) For the time course study, wild type, M1, M2, and M3 cells were seeded in T-25 flaks. At 80% confluence, cells were treated with 500 µM CuSO₄, and equal numbers of cells were removed every 24 hrs for 5 days. Collected cells were lysed in SDS-PAGE buffer, subjected to immunoblotting, and the protein was detected by anti-FLAG antibody. Each membrane was stripped and probed for actin as a loading control. (B) For the proteasome treatment study, cells were seeded and induced as above. 72 hrs post CuSO₄ inductions wild type, M2, and M3 cells were treated with 10 µM of proteasome inhibitor (MG132). Equal numbers of cells were harvested for various time points and analyzed by immunoblot.</p

Making of a <i>HeT-A</i> Gag-FLAG stable cell line.

<p>(A) Line diagram of the PMK33 vector showing important genetic markers with <i>HeT-A</i> Gag ORF1 cloned at the SmaI restriction site and a 3x FLAG tag at the C-terminus. (B) Expression of <i>HeT-A</i> Gag-FLAG in S2 cell lines as analyzed by PCR and immunoblot. GAPDH and actin serve as internal controls for PCR and immunoblots respectively. (C) Stability of expressed <i>HeT-A</i> Gag-FLAG protein in S2 cells. Cells were seeded in a T-25 flask and induced with 500 µM CuSO4. 1.0 ml cells were collected each day for a total of 6 days and analyzed by immunoblotting. (D) <i>HeT-A</i> Gag-FLAG is targeted to the nucleus. Stably transfected S2 cells were induced with CuSO₄ and 48 hr post-induction cytoplasmic and nuclear fractions were analyzed by immunoblot. (E) <i>HeT-A</i> Gag-FLAG is only detected in stably transfected S2 cells and not in S2 cells. Cytoplasmic, nuclear, and nuclei fractions from both S2 and stably transfected S2 cells were analyzed by immunoblot (1 =  cytosolic, 2 =  nuclear, and 3 =  nuclei).</p