Mutagenic and Spectroscopic Investigation of pH Dependent CooA DNA Binding

The carbon monoxide (CO) sensing heme protein, CooA, is a transcription factor which exists in several bacteria that utilize CO as an energy source. CooA positively regulates the expression of coo genes in the presence of CO such that the corresponding proteins may metabolize CO. The present studies have yielded the unexpected result that Fe(III) CooA binds DNA tightly at pH \u3c 7, deviating from all previously reported work which indicate that CooA DNA binding is initiated only when the exogenous CO effector reacts with the Fe(II) CooA heme. This observation suggests that the disruption of one or more salt bridges upon effector binding may be a critical feature of the normal CooA activation mechanism. To test this possibility, several protein variants that eliminated a selected salt bridge for the CooA homolog from Rhodospirillum rubrum were prepared via site-directed mutagenesis. Samples of these variant proteins, which were overexpressed in Escherichia coli, were then characterized by spectroscopic methods and functional assays to investigate the impact these mutations had on CooA heme coordination structure and DNA-binding activity. Results of this work are presented in light of the accepted CooA activation mechanism

The Influence of pH Variation on CooA Activity

CooA, a CO-sensing heme protein, acts as a transcriptional activator of CO-metabolizing proteins in bacteria such as Rhodospirillum rubrum and Carboxydothermus hydrogenoformans through sequence-specific DNA binding. Previous research indicated a reduced iron center and CO gas were necessary for CooA to achieve its active conformation and bind DNA. To determine if other reaction conditions facilitate CooA activation, the role of pH on CooA function was tested. Specifically, a fluorescence anisotropy assay was employed to measure possible Fe(III) CooA DNA binding from pH 3 - 12. Interestingly, CooA was observed to bind DNA without CO at acidic conditions, with optimal binding observed at pH ~3. These results are discussed in light of the normal CO-dependent activation mechanism of CooA proteins

47.4: Blue Phosphorescent Organic Light Emitting Device Stability Analysis

A model based on defect generation by exciton‐polaron annihilation interactions between the emitter and host molecules, in a blue phosphorescent OLED, is shown to fit well with experimental data. A blue PHOLED with (0.15, 0.25) chromaticity is shown to have a half‐life, from 1,000 nits, of 690 hrs.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/92134/1/1.3069766.pd

Different doses of supplemental vitamin D maintain interleukin-5 without altering skeletal muscle strength: a randomized, double-blind, placebo-controlled study in vitamin D sufficient adults

<p>Abstract</p> <p>Background</p> <p>Supplemental vitamin D modulates inflammatory cytokines and skeletal muscle function, but results are inconsistent. It is unknown if these inconsistencies are dependent on the supplemental dose of vitamin D. Therefore, the purpose of this study was to identify the influence of different doses of supplemental vitamin D on inflammatory cytokines and muscular strength in young adults.</p> <p>Methods</p> <p>Men (<it>n </it>= 15) and women (<it>n </it>= 15) received a daily placebo or vitamin D supplement (200 or 4000 IU) for 28-d during the winter. Serum 25-hydroxyvitamin D (25(OH)D), cytokine concentrations and muscular (leg) strength measurements were performed prior to and during supplementation. Statistical significance of data were assessed with a two-way (time, treatment) analysis of variance (ANOVA) with repeated measures, followed by a Tukey's Honestly Significant Difference to test multiple pairwise comparisons.</p> <p>Results</p> <p>Upon enrollment, 63% of the subjects were vitamin D sufficient (serum 25(OH)D ≥ 30 ng/ml). Serum 25(OH)D and interleukin (IL)-5 decreased (<it>P </it>< 0.05) across time in the placebo group. Supplemental vitamin D at 200 IU maintained serum 25(OH)D concentrations and increased IL-5 (<it>P </it>< 0.05). Supplemental vitamin D at 4000 IU increased (<it>P </it>< 0.05) serum 25(OH)D without altering IL-5 concentrations. Although serum 25(OH)D concentrations correlated (<it>P </it>< 0.05) with muscle strength, muscle strength was not changed by supplemental vitamin D.</p> <p>Conclusion</p> <p>In young adults who were vitamin D sufficient prior to supplementation, we conclude that a low-daily dose of supplemental vitamin D prevents serum 25(OH)D and IL-5 concentration decreases, and that muscular strength does not parallel the 25(OH)D increase induced by a high-daily dose of supplemental vitamin D. Considering that IL-5 protects against viruses and bacterial infections, these findings could have a broad physiological importance regarding the ability of vitamin D sufficiency to mediate the immune systems protection against infection.</p

Success Rate of Embolization for Type II Endoleaks at a Major Tertiary Referral Center

Objective: The rise of endovascular aneurysm repair (EVAR) as the preferred treatment for abdominal aortic aneurysm (AAA) has introduced endoleaks as a major complication following AAA repair. The objective of this study is to assess the outcomes associated with endovascular embolization of type II endoleaks after EVAR. Methods: The institutional Radiology database at our tertiary referral hospital was queried for type II endoleak during the period 2006-2018. A retrospective chart review was then carried out. Only patients who underwent intervention for isolated type 2 endoleaks were analyzed. The primary outcome was success of the endoleak repair as determined by cessation of growth (i.e., ≤5mm change in diameter over follow-up period) of the native aneurysm sac. Patient outcomes for each failure of the above criterion were also collected. Other data pertaining to the location of endoleak, type of occlusion performed, type of embolic agent used, type of endograft used for EVAR, and incidence of aneurysm rupture were collected as secondary outcomes.Results:During this period 41 patients were treated for type II endoleaks. Demographics are shown in table 1. Cessation of growth was achieved in 28/41 (68.3%) of the patients after one embolization procedure. In 13/41 (31.7%) of patients, growth of the native aneurysm sac continued. Of the patients whose aneurysms continued to grow, 61.5% (8/13) did not undergo a second embolization. The remaining 38.5% (5/13) underwent a second embolization.Patient outcomes for both of these groups are presented in table II. None of the patients were found to have ruptured their aneurysm sac during follow-up after embolization. None of gender, race, the embolization site, or method of embolization were associated with embolization failure. Conclusions: Embolization of type II endoleaks is associated with a cessation of growth in the majority of cases and seems to be protective regarding the risk of aneurysm sac rupture. Future studies and additional follow-up will be important to elucidate the most significant risk factors for expansion and/or rupture of the endovascularly repaired abdominal aneurysm.Table I: Demographics for patients with type II endoleaks who underwent endovascular embolizationVariableValue Age (years +/- sd)75.66 +/- yearsAverage follow-up (months)62.65 monthsSex (%)71.7% male28.3% femaleRace (%)77.7% white17.8% black4.4% otherInflow vessel (%)43.2% lumbar only36.4% IMA only20.5% mixEmbolization site (%)40.5% vessel only14.3% cavity only20.5% mixEmbolization type (%)66.7% coil9.5% glue23.8% mixTable II: Outcomes for patients with continued growth after embolizationThose that did no undergo further embolizationThose that underwent a second embolization 3 were found to have type III endoleak and were successfully repaired with lining of the graft.2 whose aneurysm sac ceased growing. 2 who declined further treatment. 2 whose aneurysm sac continued to grow with persistent evidence of endoleak.1 who died from non-vascular complications. 1 who was lost to follow-up. 1 who is scheduled future surgical repair. 1 who was lost to follow-up.https://scholarlycommons.henryford.com/merf2019clinres/1026/thumbnail.jp

Bhlhe40 is an essential repressor of IL-10 during Mycobacterium tuberculosis infection

The cytokine IL-10 antagonizes pathways that contro

Addressing cancer survivors\u27 cardiovascular health using the Automated Heart Health Assessment (AH-HA) EHR tool: Initial protocol and modifications to address COVID-19 challenges

BACKGROUND: The purpose of this paper is to describe the Automated Heart-Health Assessment (AH-HA) study protocol, which demonstrates an agile approach to cancer care delivery research. This study aims to assess the effect of a clinical decision support tool for cancer survivors on cardiovascular health (CVH) discussions, referrals, completed visits with primary care providers and cardiologists, and control of modifiable CVH factors and behaviors. The COVID-19 pandemic has caused widespread disruption to clinical trial accrual and operations. Studies conducted with potentially vulnerable populations, including cancer survivors, must shift towards virtual consent, data collection, and study visits to reduce risk for participants and study staff. Studies examining cancer care delivery innovations may also need to accommodate the increased use of virtual visits. METHODS/DESIGN: This group-randomized, mixed methods study will recruit 600 cancer survivors from 12 National Cancer Institute Community Oncology Research Program (NCORP) practices. Survivors at intervention sites will use the AH-HA tool with their oncology provider; survivors at usual care sites will complete routine survivorship visits. Outcomes will be measured immediately after the study visit, with follow-up at 6 and 12 months. The study was amended during the COVID-19 pandemic to allow for virtual consent, data collection, and intervention options, with the goal of minimizing participant-staff in-person contact and accommodating virtual survivorship visits. CONCLUSIONS: Changes to the study protocol and procedures allow important cancer care delivery research to continue safely during the COVID-19 pandemic and give sites and survivors flexibility to conduct study activities in-person or remotely