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Credibility, legitimacy, and diversity of knowledge are critical

The Department of Neurobiology & Behavior at Stony Brook University offers a large‐enrollment gateway undergraduate physiology course, BIO 203 ‐ Fundamentals of Biology: Cellular and Organ Physiology. BIO 203 is a required course in the biology and biochemistry majors and in the pre‐health curriculum, and undergraduates typically register for BIO 203 in their sophomore year. Since Fall 2015, BIO 203 has been offered in two formats, traditional PowerPoint‐based lectures presented twice per week (80 min / session) in a 570‐seat auditorium with individual fixed seats and a flipped, active‐learning format delivered once per week (113 min / session) in a 250‐seat auditorium with shared desks and rotating chairs. Students in both sections had equal access to prerecorded lectures and online activities (content quizzes, journal activities and thought questions). The focus of the lecture section was content delivery while the flipped section emphasized active group learning facilitated by instructors and experienced undergraduate teaching assistants. Student learning was evaluated by performance on common high‐stakes multiple choice exams. On average, students in the flipped section exhibited better performance on the common exams than students in the lecture section (Fall 2019 exam total, p < 0.05 in two sample t test and Wilcoxon rank sum test). As a result of the COVID‐19 pandemic, Stony Brook University required large‐enrollment courses to switch to online instruction starting in March 2020 and continuing through the Fall 2020 semester. The BIO 203 instructors implemented a synchronous online version of BIO 203 based on the active‐learning curriculum developed for the flipped section. In Fall 2020, this synchronous online version of BIO 203 was offered simultaneously to two sections. Students in both sections had access to the same online resources (recorded lectures, content quizzes and activities) and were evaluated using low‐stakes quizzes (administered during scheduled class time) and a common cumulative final exam (administered during a common final exam period). The primary difference between the two sections was the frequency and duration of the synchronous online meetings with instructors and teaching assistants. Section 01 (595 students) met twice a week for 80 min per session, whereas section 02 (257 students) met once a week for 130 min. A preliminary analysis of the scores on the common final exam indicates that students in section 01 (two meetings per week) performed better than students in section 02 (1 meeting per week) (p < 0.05 in two sample t test and Wilcoxon rank sum test). These preliminary findings are consistent with the hypothesis that student learning is enhanced by frequent engagement in active learning.

The health of the ocean, central to human well-being, has now reached a critical point. Most fish stocks are overexploited, climate change and increased dissolved carbon dioxide are changing ocean chemistry and disrupting species throughout food webs, and the fundamental capacity of the ocean to regulate the climate has been altered. However, key technical, organizational, and conceptual scientific barriers have prevented the identification of policy levers for sustainability and transformative action. Here, we recommend key strategies to address these challenges, including (1) stronger integration of sciences and (2) ocean-observing systems, (3) improved science-policy interfaces, (4) new partnerships supported by (5) a new ocean-climate finance system, and (6) improved ocean literacy and education to modify social norms and behaviors. Adopting these strategies could help establish ocean science as a key foundation of broader sustainability transformations.

Increasing demand for solution-oriented environmental assessments brings significant opportunities and challenges at the science–policy–society interface. Solution-oriented assessments should enable inclusive deliberative learning processes about policy alternatives and their practical consequences.

A major challenge today and into the future is to maintain or enhance beneficial contributions of nature to a good quality of life for all people. This is among the key motivations of the Intergovernmental Science-Policy Platform on Biodiversity and Ecosystem Services (IPBES), a joint global effort by governments, academia, and civil society to assess and promote knowledge of Earth's biodiversity and ecosystems and their contribution to human societies in order to inform policy formulation. One of the more recent key elements of the IPBES conceptual framework (1) is the notion of nature's contributions to people (NCP), which builds on the ecosystem service concept popularized by the Millennium Ecosystem Assessment (MA) (2). But as we detail below, NCP as defined and put into practice in IPBES differs from earlier work in several important ways. First, the NCP approach recognizes the central and pervasive role that culture plays in defining all links between people and nature. Second, use of NCP elevates, emphasizes, and operationalizes the role of indigenous and local knowledge in understanding nature's contribution to people.

In 1984, Watson and Heilman reported a patient with a partial callosal disconnection following an infarction of the anterior portion of her corpus callosum. This woman's performance on line-bisection tasks revealed "callosal disconnection neglect." The objective of this research is to reexamine this woman 34 years after her callosal disconnection to gain information about her recovery. The patient completed visual line-bisection tasks in which horizontal lines were placed in the right, left, and center hemispaces and she performed these bisections using her right or left hand. Unlike her performance 34 years ago in which each hand deviated to its ipsilateral hemispace, with greater deviation when lines were placed in the contralateral rather than ipsilateral hemispace, currently, there were no significant main effects for hand or spatial position. Thus, there were notable differences between this woman's most recent performance on the line bisection and her previous performance 34 years ago. Unlike her prior testing 34 years back, this woman's most recent performance resembled the performance of a previous tested healthy control group for whom differences in hand and hemispace were not found. It remains unclear whether her callosal disconnection neglect improved because each hemisphere learned to allocate ipsilateral spatial attention or because she learned a compensatory strategy in which she turned her body so that the lines placed in her right or left hemispace were now toward her midline.

Der Report wurde im Auftrag der Republik Nauru fur den Ban Ki-moon Klimagipfel im September 2014 in New York erstellt. Die Republik Nauru hat derzeit den Vorsitz der Allianz der kleinen Inselstaaten (Alliance of Small Island States, AOSIS) inne. In dem Report werden Prioritaten fur einen raschen Ubergang zu einer klimavertraglichen Weltwirtschaft herausgearbeitet. AOSIS ist eine Koalition von 44 niedrig gelegenen Kustenstaaten, die von den Auswirkungen des Klimawandels am starksten betroffen sind. Der Report wurde von den AOSIS-Staaten an den Generalsekretar der Vereinten Nationen ubergeben.

Background Medical student wellness has emerged as an important issue in medical education. The purpose of the present study was to obtain a comprehensive assessment of substance use, psychological distress, and help-seeking among male and female medical students in order to identify targets for continued intervention efforts. Methods Medical students from all 9 medical schools in the state of Florida were invited via e-mail and/or announcements to complete an anonymous online questionnaire assessing their well-being. Of 5053 matriculating medical students, 1137 (57.1% female) responded to the questionnaire. Descriptive statistics, t tests, and chi-square analyses were computed using SPSS 20. Results Over 70% of students acknowledged binge drinking, with men reporting higher frequency than women (χ2 = 13.90, P = .003), and 22.7% ( n = 201) reported marijuana use during medical school, with higher rates (χ2 = 9.50, P = .02) among men (27.0%, n = 99) than women (18.9%, n = 93). A significant minority of students reported nonmedical use of prescription stimulants and prescription opioids. In addition, 3.3% of male students ( n = 12) compared with 0.6% of female students ( n = 3) reported problematic drug use. Further, almost 2/3 of respondents reported decreased psychological health since beginning medical school, with women noting greater reductions (χ2 = 12.39, P = .05) and higher levels of stress (χ2 = 16.30, P = .003). Over 10% of students ( n = 102) endorsed “thoughts of committing suicide” during medical school, and 70.1% felt they would benefit from mental healthcare (79.3% of women vs. 59.6% of men; χ2 = 41.94, P < .001), although only 39.8% accessed help. Conclusions Despite efforts to address medical student wellness, students continue to report concerning levels of psychological distress, suicidal ideation, and substance use. More work is needed to effectively address medical student mental health and well-being.

Nature is perceived and valued in starkly different and oftenconflicting ways. This paper presents the rationale for theinclusive valuation of nature's contributions to people (NCP) indecision making, as well as broad methodological steps fordoing so. While developed within the context of theIntergovernmental Platform on Biodiversity and Ecosystem Services (IPBES), this approach is more widely applicable toinitiatives at the knowledge?policy interface, which require apluralistic approach to recognizing the diversity of values. Weargue that transformative practices aiming at sustainablefutures would benefit from embracing such diversity, which require recognizing and addressing power relationships across stake holder groups that hold different values on human nature relations and NCP Fil: Pascual, Unai. University Of Cambridge; Reino Unido Fil: Balvanera, Patricia. Universidad Nacional Autónoma de México; México Fil: Díaz, Sandra Myrna. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentina Fil: Pataki, György. Corvinus University of Budapest; Hungría Fil: Roth, Eva. University of Southern Denmark; Dinamarca Fil: Stenseke, Marie. University Goteborg; Suecia Fil: Watson, Robert T.. University of East Anglia; Reino Unido Fil: Basak Dessane, Esra. Project House; Turquía Fil: Islar, Mine. Lund University; Suecia Fil: Kelemen, Eszter. Environmental Social Science Research Group; Hungría Fil: Maris, Virginie. Centre d’Ecologie Fonctionnelle et Evolutive; Francia Fil: Quaas, Martin. Kiel University; Alemania Fil: Subramanian, Suneetha M. United Nations University; Japón Fil: Wittmer, Heidi. Helmholtz-Centre for Environmental Research; Alemania Fil: Adlan, Asia. University of Khartoum; Sudán Fil: Ahn, SoEun. Korea Environment Institute; Corea del Sur Fil: Al-Hafedh, Yousef S. King Abdulaziz City for Science & Technology; Arabia Saudita Fil: Amankwah, Edward. Center for Environmental Governance; Ghana Fil: Berry, Pam. University of Washington; Estados Unidos Fil: Bilgin, Adem. University of Oxford; Reino Unido Fil: Breslow, Sara J. Ministry of Forestry and Water Affairs of Turkey; Turquía Fil: Bullock, Craig. University College Dublin; Reino Unido Fil: Caceres, Daniel Mario. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias. Departamento de Desarrollo Rural; Argentina Fil: Daly-Hassen, Hamed. University of Carthage; Túnez Fil: Figueroa, Eugenio. Universidad de Chile; Chile Fil: van den Belt, Marjan. Victoria University of Wellington; Nueva Zelanda Fil: Verma, Madhu. Indian Institute of Forest Management; India Fil: Wickson, Fern. GenØk Centre for Biosafety, Siva innovasjonssenter; Noruega Fil: Yagi, Noboyuki. The University of Tokyo; Japón

Answering large-scale questions in ecology can involve time-consuming data compilation. We show how networks of undergraduate classes can make these projects more manageable and provide an authentic research experience for students. With this approach, we examined the factors associated with plant species richness in US national wildlife refuges. We found that the richness of harmful invasive plants and the richness of native plants were positively correlated in mainland refuges but negatively correlated in island refuges. Nonnative richness and invasive richness were also positively correlated with colonization pressure as indicated by nonnative richness around each refuge. Associations between refuge characteristics and invasive plants varied substantially among regions, with refuge area and habitat diversity important predictors of invasion in some regions but not in others. Our results serve to identify the refuges that are most susceptible to plant invasion and demonstrate the potential value of a new model for education and research integration.

The Florida State University College of Medicine (FSU COM) was established in 2000, the first new MD-granting medical school in the United States in over 25 years. In its brief history, the FSU COM has developed rapidly in accordance with its founding mission to meet the need for primary care physicians, especially those caring for the elderly and the underserved. The school recently received a full continuation of accreditation for the maximum period, eight years, from the Liaison Committee on Medical Education.The authors describe FSU COM's new, innovative educational program using community-based clinical training on six statewide regional campuses and two rural sites. Third- and fourth-year students are assigned to community physicians in a one-on-one clinical training model in all of the settings where physicians practice. Over 70% of student clinical training is in such settings. The authors describe how the model operates, including curricular oversight (which ensures quality and equivalence of the educational experience at all sites), the regional campus structure, administration, education program delivery during core clerkships, and assessment of students' performance. Ongoing required faculty development for all clerkship faculty is an essential feature of the training program, as is tracking of all individual student contacts through an online clinical data collection system used for evaluation of the clerkship experiences as well as research.The authors demonstrate that the school has been highly successful in implementing its mission, and that the challenge ahead is to sustain its approach to the training of future physicians.

Loss of ability of the left upper limb (LUL) to correctly produce spatial and temporal components of skilled purposeful movements was reported 34 years ago in a woman with a callosal infarction. To learn about recovery, we recently reexamined this woman. This woman was tested for ideomotor apraxia by asking her to pantomime to command and to seeing pictures of tools. Whereas she performed normally with her right upper limb, her LUL remained severely apraxic, making many spatial (postural and movement) errors. Initially, she did not reveal loss of finger-hand deftness (limb-kinetic apraxia), and when tested again with the coin rotation task, her left hand performance was normal. Without vision, she could name objects placed in her left hand but not name numbers written in this hand. Since this woman had a callosal lesion, failure to recover cannot be accounted for by left hemisphere inhibition of her right hemisphere. Although failure for her LUL to improve may have been related to not using her LUL for skilled actions, her right hemisphere was able to observe transitive actions, and this failure of her LUL to produce skilled purposeful movements suggests her right hemisphere may have not had the capacity to learn these movement representations. Without vision, her ability to recognize objects with her left hand, but not numbers written on her left palm, suggests graphesthesia may require that her left hand did not have access to movement representations important for programming these numbers when writing.

Rapid reductions in emissions of the short-lived climate forcers black carbon, tropospheric ozone and methane have been identified as an effective strategy to slow rapid warming and melting in the Arctic and other glaciated areas in the near-term, and avert abrupt, irreversible changes while strategies to reduce emissions of CO2 are implemented to limit long-term consequences. Black carbon is a component of fine particulate matter, a traditional air pollutant with significant health impacts; it has a short atmospheric residence time of about a week, thus emissions reductions provide rapid climate benefit. This article analyses the mitigation approaches that are being discussed in multilateral fora including the Arctic Council, the Convention on Long Range Transboundary Air Pollution and the International Maritime Organization. Arctic climate dynamics will affect climate globally, which calls for leadership from Arctic nations to spur faster action to abate emissions.

Limb apraxia is the loss of the ability to perform voluntary skilled movements, when this loss cannot be attributed to elemental sensorimotor deficits. Successful manual interactions with the objects in the environment require the storage of information about movement parameters. This information is stored in specific cortical modules and the correct performance of a skilled act requires interactions between these modules. Thus, apraxia can occur with degradation of these critical representations or a disconnection between modules. The goal of this paper is to define the different forms of limb apraxia and discuss how apraxia can be induced by both a deterioration of these modules as well as disconnections between these modules that form an anatomically distributed system.

It is well known that insulin receptor substrates (IRS) act as a mediator for signal transduction of insulin, insulin-like growth factors, and several cytokines. To identify proteins that interact with IRS and modulate IRS-mediated signals, we performed yeast two-hybrid screening with IRS-1 as bait. Out of 109 cDNA-positive clones identified from a human placental cDNA library, two clones encoded 53BP2, p53-binding protein 2 (53BP2S), a short form splicing variant of the apoptosis-stimulating protein of p53 that possesses Src homology region 3 domain, and ankyrin repeats domain, and had been reported to interact with p53, Bcl-2, and NF-kappaB. Interaction of 53BP2S with IRS-1 was confirmed by glutathione S-transferase pull-down and co-immunoprecipitation assays in COS-7 cells and 3T3-L1 adipocytes. The Src homology region 3 domain and ankyrin repeats domain of 53BP2S were responsible for its interaction with IRS-1, whereas the phosphotyrosine binding domain and a central domain (amino acid residues 750-861) of IRS-1 were required for its interaction with 53BP2S. In CHO-C400 cells, expression of 53BP2S reduced insulin-stimulated IRS-1 tyrosine phosphorylation with a concomitant enhancement of IRS-2 tyrosine phosphorylation. In addition, the amount of the phosphatidylinositol 3-kinase regulatory p85 subunit associated with tyrosine-phosphorylated proteins, and activation of Akt was enhanced by 53BP2S expression. Although 53BP2S also enhanced Akt activation in 3T3-L1 adipocytes, insulin-induced glucose transporter 4 translocation was markedly inhibited in accordance with reduction of insulin-induced AS160 phosphorylation. Together these data demonstrate that 53BP2S interacts and modulates the insulin signals mediated by IRSs.

The insulin granule integral membrane protein marker phogrin-green fluorescent protein was co-localized with insulin in Min6B1 beta-cell secretory granules but did not undergo plasma membrane translocation following glucose stimulation. Surprisingly, although expression of a dominant-interfering dynamin mutant (Dyn/K44A) inhibited transferrin receptor endocytosis, it had no effect on phogringreen fluorescent protein localization in the basal or secretagogue-stimulated state. By contrast, co-expression of Dyn/K44A with human growth hormone as an insulin secretory marker resulted in a marked inhibition of human growth hormone release by glucose, KCl, and a combination of multiple secretagogues. Moreover, serial pulse depolarization stimulated an increase in cell surface capacitance that was also blocked in cells expressing Dyn/K44A. Similarly, small interference RNA-mediated knockdown of dynamin resulted in marked inhibition of glucose-stimulated insulin secretion. Together, these data suggest the presence of a selective kiss and run mechanism of insulin release. Moreover, these data indicate a coupling between endocytosis and exocytosis in the regulation of beta-cell insulin secretion.

Insulin stimulates glucose uptake by promoting translocation of the Glut4 glucose transporter from intracellular storage compartments to the plasma membrane. In the absence of insulin, Glut4 is retained intracellularly, although the mechanism underlying this process remains uncertain. Using the TC10-interacting protein CIP4 as bait in a yeast two-hybrid screen, we cloned a RasGAP and VPS9 domain-containing protein, Gapex-5/RME-6. The VPS9 domain is a guanine nucleotide exchange factor for Rab31, a Rab5 subfamily GTPase implicated in trans-Golgi Network (TGN)-to-endosome trafficking. Overexpression of Rab31 blocks insulin-stimulated Glut4 translocation, whereas knockdown of Rab31 potentiates insulin-stimulated Glut4 translocation and glucose uptake. Gapex-5 is predominantly cytosolic in untreated cells; its overexpression promotes intracellular retention of Glut4 in adipocytes. Insulin recruits the CIP4/Gapex-5 complex to the plasma membrane, thus reducing Rab31 activity and permitting Glut4 vesicles to translocate to the cell surface, where Glut4 docks and fuses to transport glucose into the cell.

In muscle and adipose cells, the insulin-responsive aminopeptidase (IRAP) is localized to intracellular storage sites and undergoes insulin-dependent redistribution to the cell surface. Following expression, the newly synthesized IRAP protein traffics to the perinuclear insulin-sensitive compartment and acquires insulin sensitivity 6-9 h following biosynthesis. Knockdown of GGA1 by RNA interference prevented IRAP from entering, but not exiting, the insulin-responsive compartment. Mutation of the dileucine motif at positions 76 and 77 (EGFP-IRAP/AA(76,77)), but not the dileucine motif at positions 53 and 54, resulted in the rapid default of the reporter to the cell surface beginning at 3 h following biosynthesis. Alanine substitution of 9 residues amino- or carboxyl-terminal to LL(76,77) did not perturb basal intracellular sequestration or abrogate insulin-stimulated IRAP translocation. Moreover, a dominant interfering GGA mutant (VHS-GAT) potently inhibited insulin-stimulated translocation of EGFP-IRAP/WT but did not block the constitutive exocytotic trafficking of EGFP-IRAP/AA(76,77). In addition, the EGFP-IRAP/WT and EGFP-IRAP/AA(76,77) constructs occupied morphologically distinct tubulovesicular compartments in the perinuclear region. Taken together, these data indicate that LL(76,77) functions during the GGA-dependent sorting of newly made IRAP into the insulin-responsive storage compartment.

Upon binding and activating its cell-surface receptor, insulin triggers signaling cascades that regulate many cellular processes. Regarding glucose homeostasis, insulin suppresses hepatic glucose production and increases glucose transport into muscle and adipose tissues. At the cellular level, glucose uptake results from the insulin-stimulated translocation of the glucose transporter 4 (GLUT4) from intracellular storage sites to the plasma membrane. Although the signaling molecules that function proximal to the activated insulin receptor have been well characterized, it is not known how the distal insulin-signaling cascade interfaces with and mobilizes GLUT4-containing compartments. Recently, several candidate signaling molecules, including AS160, PIKfyve and synip, have been identified that might provide functional links between the insulin signaling cascade and GLUT4 compartments. Future work will focus on delineating the precise GLUT4 trafficking steps regulated by these molecules.

To examine the acquisition of insulin sensitivity after the initial biosynthesis of the insulin-responsive aminopeptidase (IRAP), 3T3-L1 adipocytes were transfected with an enhanced green fluorescent protein-IRAP (EGFP-IRAP) fusion protein. In the absence of insulin, IRAP was rapidly localized (1–3 h) to secretory membranes and retained in these intracellular membrane compartments with little accumulation at the plasma membrane. However, insulin was unable to induce translocation to the plasma membrane until 6–9 h after biosynthesis. This was in marked contrast to another type II membrane protein (syntaxin 3) that rapidly defaulted to the plasma membrane 3 h after expression. In parallel with the time-dependent acquisition of insulin responsiveness, the newly synthesized IRAP protein converted from a brefeldin A-sensitive to a brefeldin A-insensitive state. The initial trafficking of IRAP to the insulin-responsive compartment was independent of plasma membrane endocytosis, as expression of a dominant-interfering dynamin mutant (Dyn/K44A) inhibited transferrin receptor endocytosis but had no effect on the insulin-stimulated translocation of the newly synthesized IRAP protein.

This paper discusses key issues in the science–policy interface. It stresses the importance of linking the conservation and sustainable use of biodiversity to the Millennium Development Goals and to issues of immediate concern to policy-makers such as the economy, security and human health. It briefly discusses the process of decision-making and how the scientific and policy communities have successfully worked together on global environmental issues such as stratospheric ozone depletion and climate change, and the critical role of international assessments in providing the scientific basis for informed policy at the national and international level. The paper also discusses the drivers of global environmental change, the importance of constructing plausible futures, indicators of change, the biodiversity 2010 target and how environmental issues such as loss of biodiversity, stratospheric ozone depletion, land degradation, water pollution and climate change cannot be addressed in isolation because they are strongly interconnected and there are synergies and trade-offs among the policies, practices and technologies that are used to address these issues individually.

We have recently reported that following initial biosynthesis, the GLUT4 protein exits the Golgi apparatus and directly enters the insulin-responsive compartment(s) without transiting the plasma membrane (Watson, R. T., Khan, A. H., Furukawa, M., Hou, J. C., Li, L., Kanzaki, M., Okada, S., Kandror, K. V., and Pessin, J. E. (2004) EMBO J. 23, 2059–2070). To investigate the structural motifs involved in these initial sorting events, we have generated a variety of loss-of-function and gain-of-function GLUT4/GLUT1 chimera proteins. Substitution of the GLUT4 carboxyl-terminal domain with GLUT1 had no significant effect on the acquisition of insulin responsiveness. In contrast, substitution of either the GLUT4 amino-terminal domain or the large cytoplasmic loop between transmembrane domains 6 and 7 resulted in the rapid default of GLUT4 to the plasma membrane with blunted insulin response. Consistent with these findings, substitution of the amino-terminal, cytoplasmic loop, or carboxyl-terminal domains individually into GLUT1 backbone did not recapitulate normal GLUT4 trafficking. Similarly, dual substitutions of the GLUT1 amino and carboxyl termini with GLUT4 domains or the combination of the cytoplasmic loop plus the carboxyl terminus failed to display normal GLUT4 trafficking. However, the dual replacement of the amino terminus plus the cytoplasmic loop of GLUT4 in the GLUT1 backbone resulted in a complete restoration of normal GLUT4 trafficking. Alanine-scanning mutagenesis of the GLUT4 amino terminus demonstrated that Phe5 and Ile8 within the FQQI motif and, to a lesser extent, Asp12/Gly13 were necessary for the appropriate initial trafficking following biosynthesis. In addition, amino acids 229–271 in the large intracellular loop between transmembrane domains 6 and 7 functionally cooperated with the amino-terminal domain. These data demonstrate that initial trafficking of GLUT4 from the Golgi to the insulin-responsive GLUT4 compartment requires the functional interaction of two distinct domains.

Following biosynthesis, both GLUT1 and VSV-G proteins appear rapidly (2-3 h) at the plasma membrane, whereas GLUT4 is retained in intracellular membrane compartments and does not display any significant insulin responsiveness until 6-9 h. Surprisingly, the acquisition of insulin responsiveness did not require plasma membrane endocytosis, as expression of a dominant-interfering dynamin mutant (Dyn/K44A) had no effect on the insulin-stimulated GLUT4 translocation. Furthermore, expression of endocytosis-defective GLUT4 mutants or continuous surface labeling with an exofacial specific antibody demonstrated that GLUT4 did not transit the cell surface prior to the acquisition of insulin responsiveness. The expression of a dominant-interfering GGA mutant (VHS-GAT) had no effect on the trafficking of newly synthesized GLUT1 or VSV-G protein to the plasma membrane, but completely blocked the insulin-stimulated translocation of newly synthesized GLUT4. Furthermore, in vitro budding of GLUT4 vesicles but not GLUT1 or the transferrin receptor was inhibited by VHS-GAT. Together, these data demonstrate that following biosynthesis, GLUT4 directly sorts and traffics to the insulin-responsive storage compartment through a specific GGA-sensitive process.

Since the discovery of insulin roughly 80 yr ago, much has been learned about how target cells receive, interpret, and respond to this peptide hormone. For example, we now know that insulin activates the tyrosine kinase activity of its cell surface receptor, thereby triggering intracellular signaling cascades that regulate many cellular processes. With respect to glucose homeostasis, these include the function of insulin to suppress hepatic glucose production and to increase glucose uptake in muscle and adipose tissues, the latter resulting from the translocation of the glucose transporter 4 (GLUT4) to the cell surface membrane. Although simple in broad outline, elucidating the molecular intricacies of these receptor-signaling pathways and membrane-trafficking processes continues to challenge the creative ingenuity of scientists, and many questions remain unresolved, or even perhaps unasked. The identification and functional characterization of specific molecules required for both insulin signaling and GLUT4 vesicle trafficking remain key issues in our pursuit of developing specific therapeutic agents to treat and/or prevent this debilitating disease process. To this end, the combined efforts of numerous research groups employing a range of experimental approaches has led to a clearer molecular picture of how insulin regulates the membrane trafficking of GLUT4.

To measure students' competencies in evidence-based medicine (EBM) skills [clinical decision making using evidence from published literature (content) and in transmitting clinical information to patients (communication)] within the context of a performance-based examination (PBE).In 2002-03, under the direction of a Performance-Based Examination Oversight Committee, 16 EBM queries were developed for a pair of third-year PBEs. At the last station of the PBE, the standardized patient (SP) for that station asked a clinical EBM question relating to their "disease process." Students were asked to develop an appropriate clinical question, perform a Medline search for appropriate articles, critically appraise a complete selected article, reach a conclusion to their question, and transmit the information to the SP. Each student's clinical question, search terms, selected articles, and rationale were evaluated by faculty question-writers, clinical librarians, and the EBM course director using a five-point Likert scale, with 1 being inadequate performance and 5 being superior performance. The SP evaluated the communication skills using a checklist.Students' performances were very good, with means of 3.7 to 4.0 in each area. Agreement between the course director and station developers was good. Seventy-five percent of the students performed adequate Medline searches. Students averaged over 93% on the performance of four communication skills.The evaluation of EBM skills can be carried out during a performance-based examination. Results can assist in developing students' skills and directing curricular efforts.

Medical schools, once devoted primarily to educating medical students, have evolved into complex academic medical centers (AMCs), some of which place a greater emphasis on research and the clinical business than on educating future physicians. This occurred primarily as the result of outside forces, specifically the available revenue streams that have fostered growth. Discipline-based departments have been at the center of the governance structure of medical schools, but many AMCs now have research institutes and centers to enhance research productivity, and faculty group practices to maximize clinical revenue. Although AMCs have been successful in making scientific discoveries, developing new technologies, and providing state-of-the-art clinical care, their successes have not always been favorable to the education mission. Furthermore, the roles of departments and their chairs have not always been carefully considered; a mismatch between organizational and governance structures is occurring. In this article several suggestions are offered to help medical schools rediscover their unique reason for existence and better distinguish core missions from core businesses. Mission-based management and mission-based budgeting provide the framework for maximum success of all the missions. Specific suggestions include (1) organizing a national task force to consider optimal organizational and governance structures of modern AMCs, (2) establishing a core teaching faculty, (3) creating a matrix letter of assignment that aligns salary rates with assigned activities, (4) linking education to the provision of health care to the underinsured, and (5) forming education centers to effectively centralize governance of the education mission.

To examine the structural determinants necessary for TC10 trafficking, localization, and function in adipocytes, we generated a series of point mutations in the carboxyl-terminal targeting domain of TC10. Wild-type TC10 (TC10/WT) localized to secretory membrane compartments and caveolin-positive lipid raft microdomains at the plasma membrane. Expression of a TC10/C206S point mutant resulted in a trafficking and localization pattern that was indistinguishable from that of TC10/WT. In contrast, although TC10/C209S or the double TC10/C206,209S mutant was plasma membrane localized, it was excluded from both the secretory membrane system and the lipid raft compartments. Surprisingly, inhibition of Golgi membrane transport with brefeldin A did not prevent plasma membrane localization of TC10 or H-Ras. Moreover, inhibition of trans-Golgi network exit with a 19 degrees C temperature block did not prevent the trafficking of TC10 or H-Ras to the plasma membrane. These data demonstrate that TC10 and H-Ras can both traffic to the plasma membrane by at least two distinct transport mechanisms in adipocytes, one dependent upon intracellular membrane transport and another independent of the classical secretory membrane system. Moreover, the transport through the secretory pathway is necessary for the localization of TC10 to lipid raft microdomains at the plasma membrane.

Incubation of isolated GLUT4-containing vesicles with Xenopus oocyte extracts resulted in a guanosine 5'-[gamma-thio]triphosphate (GTP gamma S) and sodium orthovanadate stimulation of actin comet tails. The in vitro actin-based GLUT4 vesicle motility was inhibited by both latrunculin B and a dominant-interfering N-WASP mutant, N-WASP/Delta VCA. Preparations of gently sheared (broken) 3T3L1 adipocytes also displayed GTP gamma S and sodium orthovanadate stimulation of actin comet tails on GLUT4 intracellular compartments. Furthermore, insulin pretreatment of intact adipocytes prior to gently shearing also resulted in a marked increase in actin polymerization and actin comet tailing on GLUT4 vesicles. In addition, the insulin stimulation of actin comet tails was completely inhibited by Clostridum difficile toxin B, demonstrating a specific role for a Rho family member small GTP-binding protein. Expression of N-WASP/Delta VCA in intact cells had little effect on adipocyte cortical actin but partially inhibited insulin-stimulated GLUT4 translocation. Taken together, these data demonstrate that insulin can induce GLUT4 vesicle actin comet tails that are necessary for the efficient translocation of GLUT4 from intracellular storage sites to the plasma membrane.

Insulin increases glucose transport into cells of target tissues, primarily striated muscle and adipose. This is accomplished via the insulin-dependent translocation of the facilitative glucose transporter 4 (GLUT4) from intracellular storage sites to the plasma membrane. Insulin binds to the cell-surface insulin receptor and activates its intrinsic tyrosine kinase activity. The subsequent activation of phosphatidylinositol 3-kinase (PI 3-K) is well known to be necessary for the recruitment of GLUT4 to the cell surface. Both protein kinase B (PKB) and the atypical protein kinase C(lambda/zeta) (PKClambda/zeta) appear to function downstream of PI 3-K, but how these effectors influence GLUT4 translocation remains unknown. In addition, emerging evidence suggests that a second signaling cascade that functions independently of the PI 3-K pathway is also required for the insulin-dependent translocation of GLUT4. This second pathway involves the Rho-family GTP binding protein TC10, which functions within the specialized environment of lipid raft microdomains at the plasma membrane. Future work is necessary to identify the downstream effectors that link TC10, PKB, and PKClambda/zeta to GLUT4 translocation. Progress in this area will come from a better understanding of the compartmentalization of GLUT4 within the cell and of the mechanisms responsible for targeting the transporter to specialized insulin-responsive storage compartments. Furthermore, an understanding of how GLUT4 is retained within and released from these compartments will facilitate the identification of downstream signaling molecules that function proximal to the GLUT4 storage sites.

Tensilon (edrophonium chloride) is a reversible acetylcholinesterase inhibitor used in the diagnosis of myasthenia gravis, diagnosis and treatment of arrhythmias, detection of early digitalis toxicity, reversal of neuromuscular blockade, and other medical conditions. Toxicity associated with Tensilon use has appeared in the literature for decades.This review discusses the risks of Tensilon and the information practitioners should know before administering the drug. We review the literature regarding serious toxicity of this drug and offer recommendations for its safe use.A careful medication history should be taken before the administration of Tensilon. Additionally, physicians should be aware of appropriate alternative methods of diagnosis before choosing to administer Tensilon. Physicians should be aware of the clinical situations where the Tensilon test no longer is indicated.

The stimulation of glucose uptake by insulin in muscle and adipose tissue requires translocation of the GLUT4 glucose transporter protein from intracellular storage sites to the cell surface. Although the cellular dynamics of GLUT4 vesicle trafficking are well described, the signalling pathways that link the insulin receptor to GLUT4 translocation remain poorly understood. Activation of phosphatidylinositol-3-OH kinase (PI(3)K) is required for this trafficking event, but it is not sufficient to produce GLUT4 translocation. We previously described a pathway involving the insulin-stimulated tyrosine phosphorylation of Cbl, which is recruited to the insulin receptor by the adapter protein CAP. On phosphorylation, Cbl is translocated to lipid rafts. Blocking this step completely inhibits the stimulation of GLUT4 translocation by insulin. Here we show that phosphorylated Cbl recruits the CrkII-C3G complex to lipid rafts, where C3G specifically activates the small GTP-binding protein TC10. This process is independent of PI(3)K, but requires the translocation of Cbl, Crk and C3G to the lipid raft. The activation of TC10 is essential for insulin-stimulated glucose uptake and GLUT4 translocation. The TC10 pathway functions in parallel with PI(3)K to stimulate fully GLUT4 translocation in response to insulin.

Recent studies indicate that insulin stimulation of glucose transporter (GLUT)4 translocation requires at least two distinct insulin receptor–mediated signals: one leading to the activation of phosphatidylinositol 3 (PI-3) kinase and the other to the activation of the small GTP binding protein TC10. We now demonstrate that TC10 is processed through the secretory membrane trafficking system and localizes to caveolin-enriched lipid raft microdomains. Although insulin activated the wild-type TC10 protein and a TC10/H-Ras chimera that were targeted to lipid raft microdomains, it was unable to activate a TC10/K-Ras chimera that was directed to the nonlipid raft domains. Similarly, only the lipid raft–localized TC10/ H-Ras chimera inhibited GLUT4 translocation, whereas the TC10/K-Ras chimera showed no significant inhibitory activity. Furthermore, disruption of lipid raft microdomains by expression of a dominant-interfering caveolin 3 mutant (Cav3/DGV) inhibited the insulin stimulation of GLUT4 translocation and TC10 lipid raft localization and activation without affecting PI-3 kinase signaling. These data demonstrate that the insulin stimulation of GLUT4 translocation in adipocytes requires the spatial separation and distinct compartmentalization of the PI-3 kinase and TC10 signaling pathways.

To investigate the potential role of trimeric GTP-binding proteins regulating GLUT4 translocation in adipocytes, wild type and constitutively active G(q) (G(q)/Q209L), G(i) (G(i)/Q205L), and G(s) (G(s)/Q227L) alpha subunit mutants were expressed in 3T3L1 adipocytes. Although expression of neither the wild type nor G(i)/Q205L and G(s)/Q227L alpha subunit mutants had any effect on the basal or insulin-stimulated translocation of a co-expressed GLUT4-enhanced green fluorescent protein (EGFP) fusion protein, expression of G(q)/Q209L resulted in GLUT4-EGFP translocation in the absence of insulin. In contrast, microinjection of an inhibitory G(q)/G(11) alpha subunit-specific antibody but not a G(i) or G(s) alpha subunit antibody prevented insulin-stimulated endogenous GLUT4 translocation. Consistent with a required role for GTP-bound G(q)/G(11), expression of the regulators of G protein signaling (RGS4 and RGS16) also attenuated insulin-stimulated GLUT4-EGFP translocation. To assess the relationship between G(q)/G(11) function with the phosphatidylinositol 3-kinase dependent pathway, expression of a dominant-interfering p85 regulatory subunit, as well as wortmannin treatment inhibited insulin-stimulated but not G(q)/Q209L-stimulated GLUT4-EGFP translocation. Furthermore, G(q)/Q209L did not induce the in vivo accumulation of phosphatidylinositol-3,4,5-trisphosphate (PIP(3)), whereas expression of the RGS proteins did not prevent the insulin-stimulated accumulation of PIP(3). Together, these data demonstrate that insulin stimulation of GLUT4 translocation requires at least two independent signal transduction pathways, one mediated through the phosphatidylinositol 3-kinase and another through the trimeric GTP-binding proteins G(q) and/or G(11).

It has been previously reported that calmodulin plays a regulatory role in the insulin stimulation of glucose transport. To examine the basis for this observation, we examined the effect of a panel of calmodulin antagonists that demonstrated a specific inhibition of insulin-stimulated glucose transporter 4 (GLUT4) but not insulin- or platelet-derived growth factor (PDGF)-stimulated GLUT1 translocation in 3T3L1 adipocytes. These treatments had no effect on insulin receptor autophosphorylation or tyrosine phosphorylation of insulin receptor substrate 1 (IRS1). Furthermore, IRS1 or phosphotyrosine antibody immunoprecipitation of phosphatidylinositol (PI) 3-kinase activity was not affected. Despite the marked insulin and PDGF stimulation of PI 3-kinase activity, there was a near complete inhibition of protein kinase B activation. Using a fusion protein of the Grp1 pleckstrin homology (PH) domain with the enhanced green fluorescent protein, we found that the calmodulin antagonists prevented the insulin stimulation of phosphatidylinositol 3,4,5-trisphosphate [PI(3,4,5)P3] formation in vivo. Similarly, although PDGF stimulation increased PI 3-kinase activity in in vitro immunoprecipitation assays, there was also no significant formation of PI(3,4,5)P3 in vivo. These data demonstrate that calmodulin antagonists prevent insulin-stimulated GLUT4 translocation by inhibiting the in vivo production of PI(3,4,5)P3 without directly affecting IRS1- or phosphotyrosine-associated PI 3-kinase activity. This phenomenon is similar to that observed for the PDGF stimulation of 3T3L1 adipocytes.

To identify the targeting domains of syntaxin 6 responsible for its localization to the trans-Golgi network (TGN), we examined the subcellular distribution of enhanced green fluorescent protein (EGFP) epitope-tagged syntaxin 6/syntaxin 4 chimerae and syntaxin 6 truncation/deletion mutants in 3T3L1 adipocytes. Expression of EGFP-syntaxin 6 resulted in a perinuclear distribution identical to endogenous syntaxin 6 as determined both by confocal fluorescence microscopy and subcellular fractionation. Furthermore, both the endogenous and the expressed EGFP-syntaxin 6 fusion protein were localized to a brefeldin A-insensitive but okadaic acid-sensitive compartment characteristic of the TGN. In contrast, EGFP-syntaxin 6 constructs lacking the H2 domain were excluded from the TGN and were instead primarily localized to the plasma membrane. Although syntaxin 4 was localized to the plasma membrane, syntaxin 6/syntaxin 4 chimerae and syntaxin 6 truncations containing the H2 domain of syntaxin 6 were predominantly directed to the TGN. Importantly, the syntaxin 6 H2 domain fused to the transmembrane domain of syntaxin 4 was also localized to the TGN, demonstrating that the H2 domain was sufficient to confer TGN localization. In addition to the H2 domain, a tyrosine-based plasma membrane internalization signal (YGRL) was identified between the H1 and H2 domains of syntaxin 6. Deletion of this sequence resulted in the accumulation of the EGFP-syntaxin 6 reporter construct at the plasma membrane. Together, these data demonstrate that syntaxin 6 utilizes two distinct domains to drive its specific subcellular localization to the TGN.

In this paper I would like to present the case of why climate change is a serious environmental issue, which presents the nuclear industry with a major opportunity to meet the growing energy demands of the world. There is absolutely no question we need to increase the amount of energy in the world. Today there are two billion people who do not have electricity. This leads to the inefficient use of dung and woody materials as energy sources, causing significant health problems. I do believe climate change is an argument your industry can use, that we need to move away from the fossil era, to produce and use energy in very different ways from the ways we do it today. Article 2 of the UN Framework Convention on Climate Change (UNFCCC) states very simply that we need to stabilise greenhouse gas concentrations at levels below those which would cause dangerous anthropogenic perturbation of the climate system. It goes on to talk specifically about allowing eco-systems to adapt naturally, that agricultural production should be unaffected, and that we need to respond in a way which is sustainable from an economic standpoint. I think this last point is extremely important. Embedded in this is that we have to look at human health issues and water issues. The problem is that the word “dangerous” in Article 2 is a very subjective word, it is not something for the scientific community to debate. What we need to do as scientists is present to governments what would happen at different levels of greenhouse gas concentrations, and they, the governments of the world, have to decide at what level they want to stabilise. Just to put a couple of numbers in perspective, pre-industrial levels of carbon dioxide were 280 parts per million (ppm), while today they are 350 ppm. What is a safe level? Do we need to stabilise at today’s level, or twice today’s level, or three times today’s level? The European Union has argued that we need to stabilise at 550 ppm, that would be double the pre-industrial number and about another 200 ppm above today. The point is, it is not a scientific debate. It is a political debate about what Energy & Environment, Vol. 10, 1999, No. 1 19

C. Obst et al. provide a welcome opportunity to clarify the difference between environmental-economic cost-benefit analyses (such as ours) and environmental accounting exercises [such as the UN-SEEA ([ 1 ][1], [ 2 ][2]) initiative]. Accounting studies attempt to assess the total value of goods related to ecosystem services in a manner comparable to that used for market-priced goods in national accounts. A decline in the ecosystem services account over time signals a potential need to invest in underlying natural capital. However, such accounts do not indicate the most cost-effective form of that investment. Environmental economic analyses such as ours typically consider changes in value from the status quo that alternative investments provide, and identify those that yield higher value for money. The two approaches are complements rather than substitutes and serve differing but highly compatible elements of the decision-making process. A. Graham et al. criticize our use of “the value of nonmarket goods such as biodiversity.” Although we valued nonmarket greenhouse gas emissions and recreation, we explicitly did not attempt to define biodiversity in terms of economic values; instead, we applied a quantitative constraint prohibiting the degradation of biodiversity within our scenario analyses and examined the costs this would entail, finding them to be minor relative to other values. Graham et al. 's critique that we should not compare nonmarket values for ecosystem services with the market price of agricultural output ignores the fact that, as stated, we are conducting an economic analysis of marginal changes from the status quo and not attempting to assess the total value of food. In such assessments of changes, the use of market prices is standard ([ 3 ][3]). Indeed, there is an argument ([ 3 ][3]) that such analyses should subtract subsidies (including income support), which would reduce agricultural values ([ 4 ][4]). Graham et al. also question the possible increase in yields attributable to climate change. We do believe that UK farming will generally benefit from warmer temperatures but caution that [as detailed in ([ 5 ][5])] within areas of lower rainfall, increased drought could potentially reduce or even reverse these gains. We agree with R. Aspinall and P. Gregory that it would be better to consider net profits rather than farm gate prices, although again this would have reduced estimates of agricultural values. The need to link land use to its ecosystem service impacts favored our use of Agricultural Census ([ 6 ][6]) data, which omits profits. We are currently addressing this through a link to the Farm Business Survey ([ 7 ][7]) database. However, we disagree with the authors' contention that we should have included the added-value of post-farm food processing. Aside from the fact that the UK food processing industry is a major importer of non-UK produce, such an approach would be analogous to valuing timber at the price of fine furniture. It is the raw material value that is relevant here. Similarly, our analysis explicitly links agricultural land use to its impacts on the ecosystem service considered. 1. [↵][8] United Nations, European Commission, Food and Agriculture Organization of the United Nations, International Monetary Fund, Organisation for Economic Co-operation and Development, World Bank, “System of Environmental-Economic Accounting 2012: Central Framework, prepublication (white cover)” (2012); . 2. [↵][9] United Nations, European Commission, Organisation for Economic Co-operation and Development, World Bank, “System of Environmental-Economic Accounting 2012: Experimental Ecosystem Accounting, pre-publication (white cover)” (2013); [http://unstats.un.org/unsd/envaccounting/eea\_white\_cover.pdf][10]. 3. [↵][11] 1. H. M. Treasury , The Green Book: Appraisal and Evaluation in Central Government (The Stationery Office, London, 2003). 4. [↵][12] 1. I. J. Bateman, 2. A. A. Lovett, 3. J. S. Brainard , Applied Environmental Economics: A GIS Approach to Cost-Benefit Analysis (Cambridge Univ. Press, Cambridge, 2003). 5. [↵][13] 1. C. Fezzi 2. et al ., Env. Res. Econ., 10.1007/s10640-013-9663-x (2013). doi:10.1007/s10640-013-9663-x [OpenUrl][14][CrossRef][15] 6. [↵][16] Department for Environment, Food and Rural Affairs, June Agricultural Census (Edina, Manchester, UK, 2010). 7. [↵][17] Rural Business Research, “Farm Business Survey” (2013); [www.fbspartnership.co.uk][18]. [1]: #ref-1 [2]: #ref-2 [3]: #ref-3 [4]: #ref-4 [5]: #ref-5 [6]: #ref-6 [7]: #ref-7 [8]: #xref-ref-1-1 "View reference 1 in text" [9]: #xref-ref-2-1 "View reference 2 in text" [10]: http://unstats.un.org/unsd/envaccounting/eea_white_cover.pdf [11]: #xref-ref-3-1 "View reference 3 in text" [12]: #xref-ref-4-1 "View reference 4 in text" [13]: #xref-ref-5-1 "View reference 5 in text" [14]: {openurl}?query=rft.jtitle%253DEnv.%2BRes.%2BEcon.%26rft_id%253Dinfo%253Adoi%252F10.1007%252Fs10640-013-9663-x%26rft.genre%253Darticle%26rft_val_fmt%253Dinfo%253Aofi%252Ffmt%253Akev%253Amtx%253Ajournal%26ctx_ver%253DZ39.88-2004%26url_ver%253DZ39.88-2004%26url_ctx_fmt%253Dinfo%253Aofi%252Ffmt%253Akev%253Amtx%253Actx [15]: /lookup/external-ref?access_num=10.1007/s10640-013-9663-x&link_type=DOI [16]: #xref-ref-6-1 "View reference 6 in text" [17]: #xref-ref-7-1 "View reference 7 in text" [18]: http://www.fbspartnership.co.uk