Expression of recombinant proteins in the methane-producing archaeon Methanosarcina acetivorans

Recombinant protein expression is a necessary tool for the investigation of proteins in the post-genomic era. While many systems exist for recombinant protein expression in organisms of the eukaryotic and eubacterial domains, few to none are available in the Archaea. A recombinant protein expression system using the methanogenic archaeon Methanosarcina acetivorans was developed which uses the highly regulated cdh promoter and allows expression of recombinant protein with optional 6xHis protein fusions to facilitate rapid purification. A protocol for high-density mass cultivation of M. acetivorans in a stainless steel bioreactor configured as a pH-auxostat was developed. The cdh promoter and alternate promoters were analyzed in attempt to enhance expression of recombinant proteins. The protein expression system was tested on several proteins: the Methanocaldococcus jannaschii prolyl tRNA synthetase, the M. acetivorans prolyl tRNA synthetase, the Methanosarcina thermophila carbonic anhydrase, and the Dehalococcoides ethenogenes tricholorethylene dehalogenase

Barriers to Utilizing Medicaid Smoking Cessation Benefits

Introduction. Smoking is the number one preventable cause of deathin the United States. Under the Affordable Care Act, Kansas Medicaidcovers all seven FDA-approved smoking cessation therapies.However, it is estimated only 3% of Kansas Medicaid smokers usetreatment compared to the national estimate of 10%. The objectiveis to determine systemic barriers in place that prevent optimal utilizationof Medicaid smoking cessation benefits among KU MedicalCenter Internal Medicine patients. Methods. For this quality improvement project, a population of 169Kansas Medicaid smokers was identified who had been seen at the KUInternal Medicine Clinic from January 1, 2015 - February 16, 2016.Phone surveys were completed with 62 individuals about smokingstatus, interest in using smoking cessation treatment options, andawareness of Medicaid coverage of treatment. Results. Of the 62 respondents, 24 (39%) were prescribed pharmacotherapyand 41 (66%) were interested in using smoking cessationtreatment. There were eight who had quit smoking. Of the remaining54 smokers, 31 (57%) were unaware that Medicaid would coverpharmacotherapy. Of 24 participants who received a prescription forpharmacotherapy, 13 (54%) were able to fill the prescription at no costusing the Medicaid benefit. Conclusion. The majority of respondents were interested in usingsmoking cessation treatment, yet three main barriers existed to usingMedicaid smoking cessation benefits: physicians not prescribingtreatment to patients, patients not aware of Medicaid coverage, andinadequate pharmacy filling. Improved physician and patient awarenessof Medicaid coverage will facilitate more patients receivingsmoking cessation therapy and ultimately quitting smoking.KS J Med 2017;10(4):88-91

Predicting Persistent Opioid Use, Abuse, and Toxicity Among Cancer Survivors.

BackgroundAlthough opioids play a critical role in the management of cancer pain, the ongoing opioid epidemic has raised concerns regarding their persistent use and abuse. We lack data-driven tools in oncology to understand the risk of adverse opioid-related outcomes. This project seeks to identify clinical risk factors and create a risk score to help identify patients at risk of persistent opioid use and abuse.MethodsWithin a cohort of 106 732 military veteran cancer survivors diagnosed between 2000 and 2015, we determined rates of persistent posttreatment opioid use, diagnoses of opioid abuse or dependence, and admissions for opioid toxicity. A multivariable logistic regression model was used to identify patient, cancer, and treatment risk factors associated with adverse opioid-related outcomes. Predictive risk models were developed and validated using a least absolute shrinkage and selection operator regression technique.ResultsThe rate of persistent opioid use in cancer survivors was 8.3% (95% CI = 8.1% to 8.4%); the rate of opioid abuse or dependence was 2.9% (95% CI = 2.8% to 3.0%); and the rate of opioid-related admissions was 2.1% (95% CI = 2.0% to 2.2%). On multivariable analysis, several patient, demographic, and cancer and treatment factors were associated with risk of persistent opioid use. Predictive models showed a high level of discrimination when identifying individuals at risk of adverse opioid-related outcomes including persistent opioid use (area under the curve [AUC] = 0.85), future diagnoses of opioid abuse or dependence (AUC = 0.87), and admission for opioid abuse or toxicity (AUC = 0.78).ConclusionThis study demonstrates the potential to predict adverse opioid-related outcomes among cancer survivors. With further validation, personalized risk-stratification approaches could guide management when prescribing opioids in cancer patients

BCX4430 – A broad-spectrum antiviral adenosine nucleoside analog under development for the treatment of Ebola virus disease

SummaryThe adenosine nucleoside analog BCX4430 is a direct-acting antiviral drug under investigation for the treatment of serious and life-threatening infections from highly pathogenic viruses, such as the Ebola virus. Cellular kinases phosphorylate BCX4430 to a triphosphate that mimics ATP; viral RNA polymerases incorporate the drug's monophosphate nucleotide into the growing RNA chain, causing premature chain termination. BCX4430 is active in vitro against many RNA viral pathogens, including the filoviruses and emerging infectious agents such as MERS-CoV and SARS-CoV. In vivo, BCX4430 is active after intramuscular, intraperitoneal, and oral administration in a variety of experimental infections. In nonclinical studies involving lethal infections with Ebola virus, Marburg virus, Rift Valley fever virus, and Yellow Fever virus, BCX4430 has demonstrated pronounced efficacy. In experiments conducted in several models, both a reduction in the viral load and an improvement in survival were found to be related to the dose of BCX4430. A Phase 1 clinical trial of intramuscular administration of BCX4430 in healthy subjects is currently ongoing

Multi-Modal Exercise Training and Protein-Pacing Enhances Physical Performance Adaptations Independent of Growth Hormone and BDNF but May Be Dependent on IGF-1 in Exercise-Trained Men

OBJECTIVE: Protein-pacing (P; 5-6meals/day @ 2.0g/kgBW/day) and multi-mode exercise (RISE; resistance, interval, stretching, endurance) training (PRISE) improves muscular endurance, strength, power and arterial health in exercise-trained women. The current study extends these findings by examining PRISE on fitness, growth hormone (GH), insulin-like growth factor-1 (IGF-1), and brain-derived neurotrophic factor (BDNF) response, cardiometabolic health, and body composition in exercise-trained men. DESIGN: Twenty active males (\u3e4daysexercise/week) completed either: PRISE (n=11) or RISE (5-6meals/day @ 1.0g/kgBW/day; n=9) for 12weeks. Muscular strength (1-repetition maximum bench and leg press, 1-RM BP, and 1-RM LP), endurance (sit-ups, SU; push-ups, PU), power (squat jump, SJ, and bench throw, BT), flexibility (sit-and-reach, SR), aerobic performance (5km cycling time-trial, TT), GH, IGF-1, BDNF, augmentation index, (AIx), and body composition, were assessed at weeks 0 (pre) and 13 (post). RESULTS:At baseline, no differences existed between groups except for GH (RISE, 230±13 vs. PRISE, 382±59pg/ml, p CONCLUSIONS: Exercise-trained men consuming a P diet combined with multi-component exercise training (PRISE) enhance muscular power, strength, aerobic performance, and flexibility which are not likely related to GH or BDNF but possibly to IGF-1 response

Environmental Scan: Metro Regional Report

In 2010, the Extension Center for Family Development of the University of Minnesota launched a project to learn more about the current and future issues affecting families in Minnesota. During this environmental scan project, community-level interviews were conducted in 11 different regions of the state. This report summarizes the community-level interviews conducted in metro Minnesota. University of Minnesota Extension Center for Family Development staff members — Ali Shurilla, Mary Marczak, Jon Fu, and Heather Lee — assisted in the development of this report.This archival publication may not reflect current scientific knowledge or recommendations. Current information available from the University of Minnesota Extension: https://www.extension.umn.edu