Development and Implementation of a High Throughput Screen for the Human Sperm-Specific Isoform of Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDHS)

Glycolytic isozymes that are restricted to the male germline are potential targets for the development of reversible, non-hormonal male contraceptives. GAPDHS, the sperm-specific isoform of glyceraldehyde-3-phosphate dehydrogenase, is an essential enzyme for glycolysis making it an attractive target for rational drug design. Toward this goal, we have optimized and validated a high-throughput spectrophotometric assay for GAPDHS in 384-well format. The assay was stable over time and tolerant to DMSO. Whole plate validation experiments yielded Z’ values >0.8 indicating a robust assay for HTS. Two compounds were identified and confirmed from a test screen of the Prestwick collection. This assay was used to screen a diverse chemical library and identified fourteen small molecules that modulated the activity of recombinant purified GAPDHS with confirmed IC50 values ranging from 1.8 to 42 µM. These compounds may provide useful scaffolds as molecular tools to probe the role of GAPDHS in sperm motility and long term to develop potent and selective GAPDHS inhibitors leading to novel contraceptive agents

Mentoring student nurses -an update on the role and responsibilities of the mentor Word count: 2894 words

Abstract Facilitating the learning of student nurses in the work place is an integral role of the registered nurse. This article aims is to provide an overview of the role and responsibilities of the mentor in supporting pre -registration nursing students in clinical practice. The professional obligations for the mentor to meet the Nursing and Midwifery Council (NMC) standards (NMC, 2008a.) will be explored, including the on going requirements to keep up to date in mentoring practices. Some of the challenges within the role will be identified, including the importance of recognising and supporting the failing student. Recent changes to the preparation requirements for sign-off mentors will be discussed. The article concludes by presenting the potential benefits to both the individual, and to the practice placement provider

Inadequacy of cardiovascular risk factor management in chronic kidney transplantation - evidence from the FAVORIT study

Kidney transplant recipients (KTRs) have increased risk for cardiovascular disease (CVD). Our objective is to describe the prevalence of CVD risk factors applying standard criteria and use of CVD risk factor lowering medications in contemporary KTRs

The Role of Organizational Control Systems in Employees’ Organizational Trust and Performance Outcomes

This study examined how organizational control is related to employees’ organizational trust. We specifically focus on how different forms of control (process, outcome, and normative) relate to employees’ trust in their employing organizations and examine whether such trust in turn relates positively to employee job performance (task performance and organizational citizenship behavior). In addition, and in response to the recommendations of past research, we examined these relationships in a high control and compliance-based cultural context. Using data from 105 employee–supervisor dyads from professional services firms in Singapore, we find support for our hypothesized model. The implications of the results for theory and practice, and directions for future research, are discussed.Economics of Technology and Innovatio

Contribution of the land sector to a 1.5 °C world

Acknowledgements The analysis in this study was guided by the valuable feedback and recommendations of expert consultations and interviews, and we extend our gratitude to all those individuals who contributed to our research and analysis: Jeff Atkins (Virginia Commonwealth University), Jonah Busch (Earth Innovation Institute), Peter Ellis (The Nature Conservancy), Jason Funk (Center for Carbon Removal), Trisha Gopalakrishna (The Nature Conservancy), Alan Kroeger (Climate Focus), Bernice Lee (Chatham House), Donna Lee (Climate and Land Use Alliance), Simon Lewis (University College London), Guy Lomax (The Nature Conservancy), Dann Mitchell (University of Bristol), Raoni Rajão (University of Minas Gerais), Joeri Rogelj (IIASA), Carl-Friedrich Schleussner (Climate Analytics), Paul West (University of Minnesota), Graham Wynne (Prince of Wales International Sustainability Unit), Ana Yang (Children’s Investment Fund Foundation) and Dan Zarin (Climate and Land Use Alliance). A special thank you to Esther Chak and Mary-Jo Valentino (Imaginary Office) for designing the figures in this study. This work was generously supported by the Children’s Investment Fund Foundation and the authors’ institutions and funding sources.Peer reviewedPostprin

Social support and social structure

The burgeoning study of social support in relation to social stress and health would benefit from increased attention to issues of social structure. Three aspects of social relationships, all often referred to as social support, must be more clearly distinguished—(1) their existence or quantity (i.e., social integration), (2) their formal structure (i.e., social networks), and (3) their functional or behavioral content (i.e., the most precise meaning of “social support”)—and the causal relationships between the structure of social relationships (social integration and networks) and their functional content (social support) must be more clearly understood. Research and theory are needed on the determinants of social integration, networks, and support as well as their consequences for stress and health. Among potential determinants, macrosocial structures and processes particularly merit attention.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/45658/1/11206_2005_Article_BF01107897.pd

Recombinant human sperm-specific glyceraldehyde-3-phosphate dehydrogenase (GAPDHS) is expressed at high yield as an active homotetramer in baculovirus-infected insect cells

The sperm-specific glyceraldehyde-3-phosphate dehydrogenase (GAPDHS) isoform is a promising contraceptive target because it is specific to male germ cells, essential for sperm motility and male fertility, and well suited to pharmacological inhibition. However, GAPDHS is difficult to isolate from native sources and recombinant expression frequently results in high production of insoluble enzyme. We chose to use the Bac-to-Bac baculovirus-insect cell system to express a His-tagged form of human GAPDHS (Hu his-GAPDHS) lacking the proline-rich N-terminal sequence. This recombinant Hu his-GAPDHS was successfully produced in Spodoptera frugiperda 9 (Sf9) cells by infection with recombinant virus as a soluble, enzymatically active form in high yield, >35mg/L culture. Biochemical characterization of the purified enzyme by mass spectrometry and size exclusion chromatography confirmed the presence of the tetrameric form. Further characterization by peptide ion matching mass spectrometry and Edman sequencing showed that unlike the mixed tetramer forms produced in bacterial expression systems, human his-GAPDHS expressed in baculovirus-infected insect cells is homotetrameric. The ability to express and purify active human GAPDHS as homotetramers in high amounts will greatly aid in drug discovery efforts targeting this enzyme for discovery of novel contraceptives and three compounds were identified as inhibitors of Hu his-GAPDHS from a pilot screen of 1120 FDA-approved compounds

Transient Facial Nerve Paralysis (Bell's Palsy) following Intranasal Delivery of a Genetically Detoxified Mutant of Escherichia coli Heat Labile Toxin

BACKGROUND: An association was previously established between facial nerve paralysis (Bell's palsy) and intranasal administration of an inactivated influenza virosome vaccine containing an enzymatically active Escherichia coli Heat Labile Toxin (LT) adjuvant. The individual component(s) responsible for paralysis were not identified, and the vaccine was withdrawn. METHODOLOGY/PRINCIPAL FINDINGS: Subjects participating in two contemporaneous non-randomized Phase 1 clinical trials of nasal subunit vaccines against Human Immunodeficiency Virus and tuberculosis, both of which employed an enzymatically inactive non-toxic mutant LT adjuvant (LTK63), underwent active follow-up for adverse events using diary-cards and clinical examination. Two healthy subjects experienced transient peripheral facial nerve palsies 44 and 60 days after passive nasal instillation of LTK63, possibly a result of retrograde axonal transport after neuronal ganglioside binding or an inflammatory immune response, but without exaggerated immune responses to LTK63. CONCLUSIONS/SIGNIFICANCE: While the unique anatomical predisposition of the facial nerve to compression suggests nasal delivery of neuronal-binding LT-derived adjuvants is inadvisable, their continued investigation as topical or mucosal adjuvants and antigens appears warranted on the basis of longstanding safety via oral, percutaneous, and other mucosal routes

Journal of Teaching Effectiveness and Student Achievement Volume 1, Issue 1

JournalAngelo State University College EducationSupervised Field Experiences for Pre-­Service Teachers:Is it Worth the Effort? Dr. Tammy Abernathy, Dr. Ginny Beck, and Dr. Shanon Taylor………….…..5 Math Remediation?-­ Success is Possible! Dr. Deborah Banker and Dr. Stella Filizola …………………………...……………..17 Improving Pre-­Service Teacher Dispositions Dr. Marcia Bolton and Dr. Dana Reisboard ……….………………………………...24 An Investigation into the Expansive-­‐Restrictive Nature of Teachers’ Learning Situated in the Workplace Dr. Eric J. Feeney ……………………………………………………………………….………33 Using Metacognitive Awareness of Fluency to Enhance Vocabulary Dr. Teri Fowler and Dr. William Laird ………………………………………….……..44 Culturally Responsive Teaching: Increasing Involvement of Minority Students and Parents Ms. Angela Piña …………………………………………………………………………………52 Teacher Candidates’ Perceptions of Special Education Dr. S. Nina Saha-­‐Gupta, Dr. Margarita Lara, and Mr. Jeffrey House………………….60 The Teacher Preparation Initiative Dr. Yolanda Salgado, Dr. Janet A. Carter, Dr. Jeannine Hurst, and Dr. Ann Marie Smith……...…..7

Tinnitus

Tinnitus is a prevalent experience and, for those who are troubled by it, it can be debilitating. Risk factors include hearing loss, ototoxic medication, head injury and depression, and at presentation the possibility of otologic disease and of anxiety/depression should be considered. Effective drug treatments have proven elusive, though this is a vibrant theme in tinnitus research. Surgical intervention for any otological pathology associated with tinnitus may be effective for that condition, but the tinnitus may persist. Presently available treatments include the provision of hearing aids when a hearing loss is identified (even when mild or unilateral), wide band sound therapy and counselling. In some patients, cognitive behavioural therapy (CBT) is indicated though availability of tinnitus specific CBT is limited in the UK. Of these treatments the evidence base is strongest for a combination of sound therapy and CBT based counselling, though clinical trials are constrained by the heterogeneity of the tinnitus patient population. Research into mechanisms of tinnitus and effective treatments now abounds, and progress is keenly anticipated