Surface Mutation Thr34His Facilitates Purification of Haemophilus influenza Carbonic Anhydrase via Metal Affinity Chromatography

In order to pursue Haemophilus influenza carbonic anhydrase (HICA) as a potential drug target, easy and efficient purification methods must be developed. While immobilized metal affinity chromatography (IMAC) may be used, complications with polyhistidine tags is a concern. Inspired by the endogenous metal affinity of Escherichia coli β-carbonic anhydrase (ECCA), we suggest that the generation of histidine clusters on HICA’s surface will facilitate its purification by metal affinity chromatography without the potential interference of His-tags. Here we investigate the Thr34His mutation as a method to generate metal affinity in HICA. Since Thr34His is located only 5.3 Å away from His32, the two residues make a vicinal histidine pair that can interact with nickel resin. We report successful generation of Thr34His HICA mutant plasmid via site-directed mutagenesis. To obtain mutant protein for metal affinity chromatography, Thr34His HICA was overexpressed in E. coli cells and isolated as a cell lysate with a concentration of 20.2 ± 0.6 mg/mL. Metal affinity chromatography was performed on the sample, and the chromatography fractions were analyzed by SDS-PAGE in order to assess the metal affinity of the mutant. SDS-PAGE revealed that while Thr34His HICA eluted at low 10 mM and 25 mM concentrations of imidazole, 150 mM imidazole was required to fully elute the mutant. These results suggest that through the generation of surface histidine pairs, HICA can be engineered to have metal affinity and thus be easily purified via IMAC

Access to Specialist Palliative Care Services and Place of Death in Ireland

The aim of this report is to provide data, analysis and commentary to stimulate discussion on emerging trends in relation to the provision of specialist palliative care in Ireland. The report mines existing data from four key sources -- the Health Service Executive's (HSE) Minimum Data Set for Palliative Care; the National Cancer Registry; the Hospital Inpatient Enquiry, and HSE population records (2011) -- to compare and contrast administrative regions of the health service against national averages on a number of key indicators (number of hospice beds, waiting times for first assessment of patient, place of death, etc), and to examine the impact of varying levels of investment in palliative care on access to services and service activity. In particular, the report seeks to explore possible relationships between access to specialist palliative care services and place of death. It is clear from the data emerging from all four sources that there is a marked correlation between the availability or otherwise of hospice/palliative care services and where people die

Australian Judicial Review

Part I outlines the basic structure of Australia’s constitutional system and considers the source and operation of Australian judicial review. Part II examines the High Court’s approach to judicial review by considering three phases in the Court’s history. First, this Part surveys the formative years of the High Court and the development of its legalistic approach to judicial review, with particular attention given to the High Court’s earliest years, from 1903 to 1919, and Sir Owen Dixon’s tenure as Chief Justice from 1952 to 1964. Second, this Part considers the Mason Court (1987-1995), which was the first High Court to conduct its work without the oversight of the Privy Council. Widely regarded as Australia’s most activist High Court, the Mason Court took a very different view of its judicial review power, making its work of particular interest in this Article. Finally, Part II studies the contemporary High Court, the Gleeson Court, and its attempt to reign in the perceived activism of the Mason years. Examining the High Court’s constitutional jurisprudence beginning in 1998, the year of Chief Justice Gleeson’s appointment, this Article contends that the Gleeson Court’s approach to judicial review bears many hallmarks of the legalistic approach adopted by the early High Court

Physician-Assisted Suicide in Oregon: A Medical Perspective

This Article examines the Oregon Death with Dignity Act from a medical perspective. Drawing on case studies and information provided by doctors, families, and other care givers, it finds that seemingly reasonable safeguards for the care and protection of terminally ill patients written into the Oregon law are being circumvented. The problem lies primarily with the Oregon Public Health Division ( OPHD ), which is charged with monitoring the law. OPHD does not collect the information it would need to effectively monitor the law and in its actions and publications acts as the defender of the law rather than as the protector of the welfare of terminally ill patients. We make explicit suggestions for what OPHD would need to do to change that

Physician-Assisted Suicide in Oregon: A Medical Perspective

This Article examines the Oregon Death with Dignity Act from a medical perspective. Drawing on case studies and information provided by doctors, families, and other care givers, it finds that seemingly reasonable safeguards for the care and protection of terminally ill patients written into the Oregon law are being circumvented. The problem lies primarily with the Oregon Public Health Division ( OPHD ), which is charged with monitoring the law. OPHD does not collect the information it would need to effectively monitor the law and in its actions and publications acts as the defender of the law rather than as the protector of the welfare of terminally ill patients. We make explicit suggestions for what OPHD would need to do to change that

The Impact of Parental Marital Disruption on Children's Performance in School

Although it is well-documented that children from divorced or single-parent families have lower performance in school, little is known about whether the timing of divorce is linked to a decline in school performance. In this paper, we examine whether and how the timing of change in parents' marital status influences the odds that a child is held back a grade in school using the Baltimore Parenthood Study, a thirty-year longitudinal study that has tracked the lives of inner-city families. Using logistic regression and hazard analysis, we show that the odds that a child is held back in school increase dramatically around the period of marital dissolution

Use of antiepileptic drugs and lipid-lowering agents in the United States.

INTRODUCTION: The extent to which enzyme-inducing antiepileptic drugs (EIAEDs) are used as first-line treatment in the United States remains unknown. Studies suggest that EIAEDs produce elevation of serum lipids, which could require additional treatment. We assessed the current use of EIAED in monotherapy for epilepsy in the U.S., as well as the correlation between the use of EIAEDs and subsequent new prescriptions for HMG-CoA reductase inhibitors ( statins ) for hyperlipidemia. METHODS: We queried the MarketScan® databases between July 2009 and January 2013, covering 66million patients with commercial or supplemental Medicare insurance. We identified individuals who had a diagnosis of seizures, continuous enrollment in the database from 6months prior to 24months after the epilepsy diagnosis, no utilization of an AED or a statin prior to that diagnosis, and at least 1 new AED prescription. We tabulated the fraction of subjects who were prescribed EIAEDs (phenytoin, carbamazepine, or barbiturates) and those prescribed all other AEDs. Rates of new statin prescription between 1 and 24months after AED prescription were assessed among the two groups, restricted to those with no prior history of vascular disease who had lipid serology obtained subsequent to the new AED prescription. RESULTS: Of the 11,893 patients with newly treated epilepsy, 2425 (20.4%) were started on an EIAED, and 9468 (79.6%) were started on a noninducing AED. There was a consistent and significant trend for EIAEDs to be increasingly prescribed with increasing age (p CONCLUSIONS: Enzyme-inducing antiepileptic drug prescription for epilepsy appears to increase with increasing age in the U.S. despite the absence of a cogent rationale for this practice, suggesting a failure to appreciate the complications of EIAED therapy among U.S. physicians. Statins were more often prescribed to those newly treated with EIAEDs compared with those given noninducing AEDs. These preliminary data provide further evidence suggesting that EIAEDs elevate lipids in a clinically meaningful manner

A METHODOLOGY FOR CLASSIFYING THE COMPLEXITY OF EXPERT SYSTEMS: A PILOT STUDY

The focus of this paper is to present a classification methodology for evaluating the complexity of expert systems. Complexity in the area of expert systems consists of two basic dimensions: the complexity of the underlying knowledge residing with the experts and the complexity of the technology incorporated into a given system. The classification methodology was developed and tested for its ability to accurately differentiate expert systems with a pilot sample of six expert systems. Using this approach provides a basis for managers to assess the complexity of a particular expert system and thereby assist in planning the scope of the development and implementation effort and the fit of a particular project with the firm\u27s internal resources and the needs of its competitive environment

Direct healthcare costs of hip, vertebral, and non-hip, non-vertebral fractures.

Limited data exist regarding the cost of non-hip, non-vertebral (NHNV) fractures. Although NHNV fractures may be less expensive than hip and vertebral fractures, they have a higher incidence rate. The objective of this study was to quantify first-year healthcare costs of hip, vertebral, and NHNV fractures. This was a claims-based retrospective analysis using a case-control design among patients with commercial insurance and Medicare employer-based supplemental coverage. Patients were \u3e or =50 years old with a closed hip, vertebral, or NHNV fracture between 7/1/2001 and 12/31/2004, and continuous enrollment 6 months prior to and 12 months after the index fracture. Adjusted mean first-year healthcare costs associated with these fractures were determined. Six cohorts were identified. Patients 50-64 years: NHNV (n=27,424), vertebral (n=3386) and hip (n=2423); patients \u3e or =65 years: NHNV (n=40,960), vertebral (n=11,751) and hip (n=21,504). The ratio of NHNV to hip fractures was 11:1 in the 50-64 cohort and 2:1 in the \u3e or =65 cohort. Adjusted mean first-year costs associated with hip, vertebral, and NHNV fractures were $26,545,$14,977, and $9183 for the 50-64 age cohort, and$15,196, $6701, and$6106 for patients \u3e or =65 years. After taking prevalence rate into account, the proportion of the total fracture costs accounted for by NHNV, hip, and vertebral fractures were 66%, 21% and 13% for the 50-64 age cohort, and 36%, 52% and 12% for the \u3e or =65 age cohort. Limitations included the exclusion of the uninsured and those covered by Medicaid or military-based insurance programs. The results of this study demonstrate that osteoporotic fractures are associated with significant costs. Although NHNV fractures have a lower per-patient cost than hip or vertebral fractures, their total first-year cost is greater for those 50-64 because of their higher prevalence

An investigation of screening for medical referral by Physical Therapists

Screening for medical referral is essential to physical therapist practice; however, no studies have examined medical screening across physical therapy practice settings. The purpose of this study was to determine if physical therapists in a variety of practice settings, given brief clinical vignettes, would screen for medical referral in a similar manner. The Delphi Technique was implemented to establish content validity, although only 2 vignettes reached 100% consensus by the expert panel. After reviewing the vignettes, physical therapist subjects determined if they would provide intervention, provide intervention and refer, or refer before intervention. To determine which variables were associated with identified appropriate decisions in each category, 4 sets of logistic regressions were performed. A random sample of APTA members in targeted sections (n=214) completed the survey. Physical therapists with more than 23 years experience were 7 times more likely to make similar management decisions in noncritical vignettes then those with 11 years or less. Half of the participants practiced in outpatient practice settings. In critical medical vignettes, 86% of physical therapists made similar decisions. Physical therapists in this study made identified appropriate management decisions for 90% of the vignettes. Board certification, practice setting and highest earned degree were not found to be statistically significant. It is imperative not only to ensure that the client is appropriate for physical therapy intervention, but it is also vital to recognize and identify signs and symptoms that warrant evaluation by other health care providers