Callosal disconnection neglect: reassessment after 34 years.

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.

Callosal apraxia: a 34-year follow-up study.

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.

Gapex-5, a Rab31 guanine nucleotide exchange factor that regulates Glut4 trafficking in adipocytes.

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; 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.

Bridging the GAP between insulin signaling and GLUT4 translocation.

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.

Do we need a new science-policy interface for food systems?

Credibility, legitimacy, and diversity of knowledge are critical.

The C-terminus of GLUT4 targets the transporter to the perinuclear compartment but not to the insulin-responsive vesicles.

Postprandial blood glucose clearance is mediated by GLUT4 (glucose transporter 4) which is translocated from an intracellular storage pool to the plasma membrane in response to insulin. The nature of the intracellular storage pool of GLUT4 is not well understood. Immunofluorescence staining shows that, under basal conditions, the major population of GLUT4 resides in the perinuclear compartment. At the same time, biochemical fractionation reveals that GLUT4 is localized in IRVs (insulin-responsive vesicles). The relationship between the perinuclear GLUT4 compartment and the IRVs is not known. In the present study, we have exchanged the C-termini of GLUT4 and cellugyrin, another vesicular protein that is not localized in the IRVs and has no insulin response. We have found that GLUT4 with the cellugyrin C-terminus loses its specific perinuclear localization, whereas cellugyrin with the GLUT4 C-terminus acquires perinuclear localization and becomes co-localized with GLUT4. This, however, is not sufficient for the effective entry of the latter chimaera into the IRVs as only a small fraction of cellugyrin with the GLUT4 C-terminus is targeted to the IRVs and is translocated to the plasma membrane in response to insulin stimulation. We suggest that the perinuclear GLUT4 storage compartment comprises the IRVs and the donor membranes from which the IRVs originate. The C-terminus of GLUT4 is required for protein targeting to the perinuclear donor membranes, but not to the IRVs.

Regulated membrane trafficking of the insulin-responsive glucose transporter 4 in adipocytes.

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.

GLUT4 translocation: the last 200 nanometers.

Insulin regulates circulating glucose levels by suppressing hepatic glucose production and increasing glucose transport into muscle and adipose tissues. Defects in these processes are associated with elevated vascular glucose levels and can lead to increased risk for the development of Type 2 diabetes mellitus and its associated disease complications. At the cellular level, insulin stimulates glucose uptake by inducing the translocation of the glucose transporter 4 (GLUT4) from intracellular storage sites to the plasma membrane, where the transporter facilitates the diffusion of glucose into striated muscle and adipocytes. Although the immediate downstream molecules that function proximal to the activated insulin receptor have been relatively well-characterized, it remains unknown how the distal insulin-signaling cascade interfaces with and recruits GLUT4 to the cell surface. New biochemical assays and imaging techniques, however, have focused attention on the plasma membrane as a potential target of insulin action leading to GLUT4 translocation. Indeed, it now appears that insulin specifically regulates the docking and/or fusion of GLUT4-vesicles with the plasma membrane. Future work will focus on identifying the key insulin targets that regulate the GLUT4 docking/fusion processes.

The disconnection apraxias.

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.

The exocytotic trafficking of TC10 occurs through both classical and nonclassical secretory transport pathways in 3T3L1 adipocytes.

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.

Small GTP-binding protein TC10 differentially regulates two distinct populations of filamentous actin in 3T3L1 adipocytes.

TC10 is a member of the Rho family of small GTP-binding proteins that has previously been implicated in the regulation of insulin-stimulated GLUT4 translocation in adipocytes. In a manner similar to Cdc42-stimulated actin-based motility, we have observed that constitutively active TC10 (TC10/Q75L) can induce actin comet tails in Xenopus oocyte extracts in vitro and extensive actin polymerization in the perinuclear region when expressed in 3T3L1 adipocytes. In contrast, expression of TC10/Q75L completely disrupted adipocyte cortical actin, which was specific for TC10, because expression of constitutively active Cdc42 was without effect. The effect of TC10/Q75L to disrupt cortical actin was abrogated after deletion of the amino terminal extension (DeltaN-TC10/Q75L), whereas this deletion retained the ability to induce perinuclear actin polymerization. In addition, alteration of perinuclear actin by expression of TC10/Q75L, a dominant-interfering TC10/T31N mutant or a mutant N-WASP protein (N-WASP/DeltaVCA) reduced the rate of VSV G protein trafficking to the plasma membrane. Furthermore, TC10 directly bound to Golgi COPI coat proteins through a dilysine motif in the carboxyl terminal domain consistent with a role for TC10 regulating actin polymerization on membrane transport vesicles. Together, these data demonstrate that TC10 can differentially regulate two types of filamentous actin in adipocytes dependent on distinct functional domains and its subcellular compartmentalization.

Effect of background motion on line bisection performance in normal subjects.

Previous studies have demonstrated that optokinetic stimulation (OKS) influences line bisection (LB) performance in normal subjects and patients with hemispatial neglect. Since subjects were required to attend to stationary targets on a moving background, prior experimental designs might have induced an illusion of target motion or induced motion (IM) in a direction opposite the background. The current study tested whether the IM affects LB performance in normal subjects and how the speed of targets also influences LB. Thirty-two right-handed normal volunteers (aged 28.0 +/- 5.3 years) were asked to bisect stationary lines with a background of horizontal OKS. These stimuli were generated by computer displayed on a large screen via a beam projector. The OKS was varied according to direction (leftward or rightward) and speed (9.4 degrees/sec or 56.1 degrees/sec), producing 4 different experimental conditions. Mean bisection errors in all conditions were compared with a control condition with no background OKS. For each condition, subjects rated the degree of IM on a 5 point scale. With fast rate OKS, subjects reported minimal IM and LB errors were in the same direction as background motion, a finding that replicates previous studies. Conversely, the slow OKS rate caused subjects to report IM and resulted in deviation of the bisection mark in a direction opposite the background OKS. While this discrepancy between the slow and fast OKS conditions might be related to motion illusion, we did not find a direct correlation between the degree of IM and bisection errors and thus reasons for these results remain unexplained.

Rediscovering the medical school.

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.

Evaluating evidence-based medicine skills during a performance-based examination.

PURPOSE: 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). METHOD: 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. RESULTS: 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. CONCLUSION: 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.

Environmental threats, mitigation strategies and high-mountain areas.

High-mountain areas suffer from increasing environmental threats. The causes are often global in dimension but lead to specific impacts under conditions of steep and/or high-altitude terrain with strong effects from snow and ice. This paper presents a global perspective, focussing primarily on observed and projected changes in climate and then goes on to discuss key messages of greatest relevance to the highest belts of mountain regions. The paper finishes with a brief discussion of mitigation strategies.

Bringing ecosystem services into economic decision-making: land use in the United Kingdom.

Landscapes generate a wide range of valuable ecosystem services, yet land-use decisions often ignore the value of these services. Using the example of the United Kingdom, we show the significance of land-use change not only for agricultural production but also for emissions and sequestration of greenhouse gases, open-access recreational visits, urban green space, and wild-species diversity. We use spatially explicit models in conjunction with valuation methods to estimate comparable economic values for these services, taking account of climate change impacts. We show that, although decisions that focus solely on agriculture reduce overall ecosystem service values, highly significant value increases can be obtained from targeted planning by incorporating all potential services and their values and that this approach also conserves wild-species diversity.