C-reactive protein is associated with cognitive performance in a large cohort of euthymic patients with bipolar disorder.

Data support the notion that 40-60% of patients with bipolar disorder (BD) have neurocognitive deficits. It is increasingly accepted that functioning in BD is negatively impacted by these deficits, yet they have not been a successful target for treatment. The biomarkers that predict cognitive deficits in BD are largely unknown, however recent evidence suggests that inflammation may be associated with poorer cognitive outcomes in BD. We measured C-reactive protein (CRP), a marker of systemic inflammation and risk of inflammatory disease, in 222 euthymic BD patients and 52 healthy controls. Within the patient sample, using multivariate analyses of covariance (MANCOVA) we compared cognitive performance of those with high CRP (≥5 mg/L) versus the remaining subjects (<5 mg/L) on a battery of cognitive tests. We evaluated relationships with several other relevant clinical features. We also examined the role of CRP in cognitive decline using a proxy cognitive decline metric, defined as the difference between premorbid and current IQ estimates, in a logistic regression analysis. Approximately 80% of our sample were BD-I, and the remainder were BD-II and 42.6% of our sample had a history of psychosis. We found a statistically significant effect of CRP on cognitive performance on a broad range of tests; participants with CRP ≥ 5 mg/L had worse performance on several measures of executive functioning, MATRICS processing speed and MATRICS reasoning and problem solving relative to those with lower CRP. We also identified CRP as a significant positive predictor of proxy cognitive decline. Our results indicate that elevated CRP is associated with a broad cognitive dysfunction in affectively remitted BD patients. These results may point to a subgroup of patients who might benefit from treatments to reduce inflammation.

TNF-α and its soluble receptors mediate the relationship between prior severe mood episodes and cognitive dysfunction in euthymic bipolar disorder.

BACKGROUND: Bipolar disorder (BD) is one of the most disabling mental health conditions in the world. Symptoms of cognitive impairment in BD contribute directly to occupational and social deficiencies and are very difficult to treat. Converging evidence suggests that BD patients have increased peripheral markers of inflammation. The hypothesis of neuroprogression in BD postulates that cognitive deficits develop over the course of the illness and are influenced by prior severe mood episodes, leading to wear-and-tear on the brain- however, there exists a paucity of data statistically testing a mediating role of immune molecules in cognitive dysfunction in BD. METHODS: This is a cross-sectional study. We measured serum levels of tumor necrosis factor alpha (TNF-α), and soluble (s) TNF receptors one and two (sTNF-R1 and sTNF-R2) in 219 euthymic BD patients and 52 Healthy Controls (HCs). Structural equation modeling (SEM) was used for the primary purpose of assessing whether TNF markers (measured by the multiple indicators TNF-α, sTNF-R1 and sTNF-R2) mediate the effect or number of prior severe mood episodes (number of prior psychiatric hospitalizations) on cognitive performance. RESULTS: BD and HC groups did not differ on circulating levels of TNF molecules in the present study. However, we found higher sTNF-R1 concentration in 'late-stage' BD illness (>1 prior psychiatric hospitalization) compared to those in early stage illness. In the subsequent SEM, we found that the model fits the data acceptably (Chi-square = 49.2, p = 0.3), and had a 'close fit' (RMSEA = 0.02, PCLOSE = 0.9). Holding covariates constant (age, sex, premorbid IQ, education, and race), we found that the standardized indirect effect was significant, p = 0.015, 90%CI [-0.07, -0.01], indicating that the estimated model was consistent with peripheral TNF markers partially mediating a causal effect of severe mood episodes on executive function. CONCLUSIONS: Our results indicate that circulating levels of TNF molecules partially mediate the relationship between prior severe mood episodes and executive function in BD. These results may implicate TNF variables in the neuroprogressive course of BD and could point to novel interventions for cognition.

Neurocognitive subtypes in patients with bipolar disorder and their unaffected siblings.

BACKGROUND: Our previous work revealed substantial heterogeneity in the cognitive profile of bipolar disorder (BD) due to the presence of three underlying cognitive subgroups characterized as: globally impaired, selectively impaired, or cognitively intact. In an effort to determine whether these subgroups are differentially related to genetic risk for the illness, we investigated whether cognitive deficits were more pronounced in unaffected siblings (UAS) of BD probands within identified clusters. METHODS: Cluster analysis was used to identify cognitive clusters in BD (N = 60). UAS (N = 49) were classified into groups according to their proband sibling's cluster assignment; comparisons were made across all clusters and healthy controls (HCs; N = 71). RESULTS: Three cognitive clusters in BD emerged: a globally impaired (36.7%), a selectively impaired (30%), and a cognitively intact cluster (33.3%). UAS showed a qualitatively similar pattern to their BD siblings; UAS of the globally impaired BD cluster showed verbal memory and general cognitive impairments relative to HCs. In contrast, UAS of the other two clusters did not differ from HCs. CONCLUSIONS: This study corroborates findings from prior work regarding the presence of cognitive heterogeneity in BD. UAS of subjects in the globally impaired BD cluster presented with a qualitatively similar cognitive profile to their siblings and performed worse than all other BD clusters and UAS groups. This suggests that inherited risk factors may be contributing to cognitive deficits more notably in one subgroup of patients with BD, pointing toward differential causes of cognitive deficits in discrete subgroups of patients with the disorder.

Empirical evidence for discrete neurocognitive subgroups in bipolar disorder: clinical implications.

BACKGROUND: Recent data suggest trait-like neurocognitive impairments in bipolar disorder (BPD), with deficits about 1 s.d. below average, less severe than deficits noted in schizophrenia. The frequency of significant impairment in BPD is approximately 60%, with 40% of patients characterized as cognitively spared. This contrasts with a more homogeneous presentation in schizophrenia. It is not understood why some BPD patients develop deficits while others do not. METHOD: A total of 136 patients with BPD completed the MATRICS Consensus Cognitive Battery and data were entered into hierarchical cluster analyses to: (1) determine the optimal number of clusters (subgroups) that fit the sample; and (2) assign subjects to a specific cluster based on individual profiles. We then compared subgroups on several clinical factors and real-world community functioning. RESULTS: Three distinct neurocognitive subgroups were found: (1) an intact group with performance comparable with healthy controls on all domains but with superior social cognition; (2) a selective impairment group with moderate deficits on processing speed, attention, verbal learning and social cognition and normal functioning in other domains; and (3) a global impairment group with severe deficits across all cognitive domains comparable with deficits in schizophrenia. CONCLUSIONS: These results suggest the presence of multiple cognitive subgroups in BPD with unique profiles and begin to address the relationships between these subgroups, several clinical factors and functional outcome. Next steps will include using these data to help guide future efforts to target these disabling symptoms with treatment.

The effect of dysmenorrhea severity and interference on reactions to experimentally-induced pain.

Introduction: Dysmenorrhea is associated with increased risk of chronic pain and hyperalgesia. Menstruating individuals with dysmenorrhea are more likely to have elevated pain reactivity when experiencing experimental pain, than those without. However, no study has examined intragroup differences in reactions to experimentally induced pain for individuals with dysmenorrhea. The main aim of this study was to examine the relative roles of dysmenorrhea severity and interference in the experience of experimentally-induced pain. Methods: Participants were 120 menstruating individuals involved in a larger research study examining the influence of expectations on experimentally-induced pain. As part of the study, participants completed an online questionnaire regarding demographic and menstrual information and participated in a cold pressor task. Participants were randomized into four groups based on the manipulation of two independent variables: (1) high vs. low expectations about pain severity (pain-expectations); (2) and high vs. low expectations about one's pain tolerance (self-expectations). Participants verbally rated their pain severity throughout the cold pressor task using a 0-10 scale. Regression analyses were conducted examining the relationships between dysmenorrhea experience (i.e., average severity and interference) and cold pressor data [pain severity ratings and pain tolerance (i.e., total time in the cold pressor)], controlling for the manipulated expectations and age. Then, moderation analyses were conducted examining expectation group differences. Results: When controlling for manipulated expectations and age, dysmenorrhea severity significantly predicted initial pain severity rating (p = 0.022) but did not predict final pain severity rating (p = 0.263) or pain tolerance (p = 0.120). Dysmenorrhea interference did not predict initial pain severity rating (p = 0.106), final pain severity rating (p = 0.134), or pain tolerance (p = 0.360). A moderation analysis indicated that the relationship between dysmenorrhea severity and initial pain severity rating was not moderated by pain-expectations, χ 2(1) = 0.412, p = 0.521. Discussion: During an experimentally-induced pain task, dysmenorrhea severity but not interference predicted initial pain severity rating, such that higher levels of dysmenorrhea severity predicted greater initial pain severity rating. This suggests individuals with more severe dysmenorrhea pain may experience greater initial sensitivity to pain and be at risk for increased sensitivity to acute pain and potentially the development of chronic pain.

Dependence of volatile droplet lifetime on the hydrophobicity of the substrate.

In this Article, we demonstrate the dependence of the lifetime of a volatile droplet on the hydrophobicity of the substrate. Ethanol droplets placed on the molecularly smooth surfaces of three polymers, applied to substrates by spin-coating, showed distinct types of behavior depending on the hydrophobicity of the latter. High contact angles, θ, lead to fairly regular recession of the triple line during liquid evaporation at essentially constant θ, whereas low contact angle caused pinning, θ decreasing with time. The latter case leads to shorter drop lifetimes.

International perspectives on radiography practice education.

INTRODUCTION: The radiography profession is built upon strong educational foundations which help ensure graduate radiographers have the required knowledge, skills, and competence to practise safely and effectively. Changing clinical practices, service needs, technological developments, regulatory changes, together with our growing professional evidence-base, all contribute to the need for our curricula to responsive and continually reviewed and enhanced. This study aims to explore similarities and differences in training curricula and follows a 2012 global survey on radiography education and more recent surveys undertaken by the European Federation of Radiographer Societies (EFRS). METHODS: An online questionnaire, based on previous EFRS education and clinical education surveys, which comprised of open and closed questions and consisted of sections designed to ascertain data on: type, level and duration of education programmes leading to an initial or pre-registration qualification in radiography/medical radiation practice, pre-clinical skill development and clinical placement within programmes. The survey was distributed via social media channels and through an international network of professional societies. Descriptive statistics are reported for most analyses while open questions were analysed thematically. RESULTS: Responses were received from 79 individuals from 28 identified countries across four continents. This represented a total of 121 different pre-registration/entry level programmes offered across these institutions. While dedicated diagnostic radiography programmes were most common (42/121), almost one-third of programmes (40/121) offered two or more areas of specialisation within the curriculum. The average of total hours for clinical placement were 1397 h for diagnostic radiography programmes; 1300 h for radiation therapy programmes; 1025 h for nuclear medicine programmes; and 1134 h for combined specialisation programmes, respectively. Institutions provided a range of physical and virtual systems to support pre-clinical skills development. CONCLUSION: Around the world, radiography programmes vary considerably in terms of their level, duration, programme type, pre-clinical and clinical training, use of simulation, and also in terms of class sizes, student/staff ratios, and graduate employment prospects. The ability of graduates to work independently in areas covered within their programmes varied considerably. While some changes around simulation use were evident, given the impact of COVID-19 it would be beneficial for future research to investigate if pre-clinical and clinical education hours or use of simulation resources has changed due to the pandemic. IMPLICATIONS FOR PRACTICE: The heterogeneity that exists between radiography programmes presents a significant challenge in terms of the mutual recognition of qualifications and the international movement of the radiographer workforce.

The global impact of the COVID-19 pandemic on clinical radiography practice: A systematic literature review and recommendations for future services planning.

INTRODUCTION: Worldwide, reports and experiences indicate that there has been extensive re-organisation within diagnostic imaging and radiotherapy departments in response to the COVID-19 pandemic. This was necessary due to changes in workload and working practice guidelines that have evolved during the pandemic. This review provides a comprehensive summary of the global impact of the COVID-19 pandemic on radiography practice, service delivery and workforce wellbeing. METHODS: A systematic review methodology was adopted to obtain data from primary studies of qualitative, quantitative, and mixed methods designs from databases (PubMed, Science Direct, Cumulative Index of Nursing and Allied Health Literature [CINAHL], and SCOPUS: all 2020 to present). The included articles were subjected to information extraction and results-based convergent synthesis. RESULTS: The electronic database search yielded 10,420 articles after removal of duplicates. Of these, 31 articles met the final inclusion criteria with some (n = 8) fully focussed on radiotherapy workforce and service delivery. The pandemic impact on radiography practice is broadly themed around: training, communication, and information dissemination; infrastructure, technology, and clinical workflow; and workforce mental health and well-being. CONCLUSION: Globally, most radiographers received inadequate training for managing COVID-19 patients during the initial acute phase of the pandemic. Additionally, there were significant changes to clinical practice, working patterns and perceived increase in workload due to surges in COVID-19 patients and the consequent strict adherence to new infection protocols. These changes, coupled with fear emanating from the increased risk of the workforce to contracting the infection, contributed to anxiety and workplace-related stress during the pandemic. IMPLICATIONS FOR PRACTICE: Local pandemic response strategies must be appropriately developed from standard protocols in readiness for safe clinical practice and well-being management training of practitioners.

Emotional processing subtypes in bipolar disorder: A cluster analysis.

BACKGROUND: Evidence regarding the performance of Bipolar Disorder patients (BD) on Emotional Processing (EP) is conflicting, suggesting that heterogeneity within this population may exist. It is not completely understood if this impacts on clinical presentation and functional outcomes. METHODS: A total of 212 BD patients were recruited. Patients underwent MATRICS Consensus Cognitive Battery as well as a clinical evaluation to detect premorbid traits, comorbidities and clinical features. Performance on each basic emotion on the Emotional Recognition Task (ERT) and Reading the Mind in the Eyes Test were entered into hierarchical cluster analyses in order to determine the number of clusters and to assign subjects to specific clusters. We then compared subgroups on clinical factors and real-world community functioning. RESULTS: No differences between BD patients as a group and controls were found in EP performance. Two clusters of BD patients were found, one with "intact" performance (71.2%) that performed as healthy controls (HC) and other with "impaired" performance (28.8%) performing worse than HC and schizophrenic patients on basic emotion recognition. Patients in the "impaired group" presented higher rates of childhood trauma, schizotypal traits, lower premorbid IQ and education, poor psychosocial functioning and cognitive performance. LIMITATIONS: Cross-sectional data which limits our ability to infer directionality of our findings. CONCLUSION: These results suggest the presence of two subgroups regarding EP performance with unique clinical and neurodevelopmental profiles associated. Next steps will include using these data to identify a homogeneous group of patients to target these disabling symptoms with treatment.

The Mortality After Release from Incarceration Consortium (MARIC): Protocol for a multi-national, individual participant data meta-analysis.

INTRODUCTION: More than 30 million adults are released from incarceration globally each year. Many experience complex physical and mental health problems, and are at markedly increased risk of preventable mortality. Despite this, evidence regarding the global epidemiology of mortality following release from incarceration is insufficient to inform the development of targeted, evidence-based responses. Many previous studies have suffered from inadequate power and poor precision, and even large studies have limited capacity to disaggregate data by specific causes of death, sub-populations or time since release to answer questions of clinical and public health relevance. OBJECTIVES: To comprehensively document the incidence, timing, causes and risk factors for mortality in adults released from prison. METHODS: We created the Mortality After Release from Incarceration Consortium (MARIC), a multi-disciplinary collaboration representing 29 cohorts of adults who have experienced incarceration from 11 countries. Findings across cohorts will be analysed using a two-step, individual participant data meta-analysis methodology. RESULTS: The combined sample includes 1,337,993 individuals (89% male), with 75,795 deaths recorded over 9,191,393 person-years of follow-up. CONCLUSIONS: The consortium represents an important advancement in the field, bringing international attention to this problem. It will provide internationally relevant evidence to guide policymakers and clinicians in reducing preventable deaths in this marginalized population. KEY WORDS: Mortality; incarceration; prison; release; individual participant data meta-analysis; consortium; cohort.

Alexithymia predicts poorer social and everyday functioning in schizophrenia and bipolar disorder.

Alexithymia, or the inability to identify and describe one's emotions, is significantly higher in bipolar disorder (BD) and schizophrenia (SZ), compared to healthy controls (HC). Alexithymia has also been observed to predict psychosocial functioning in SZ. We investigated whether alexithymia predicted social and everyday functioning in BD, as well as transdiagnostically in HC, BD, and SZ patients. 56 BD, 45 SZ, and 50 HC were administered and compared on tests measuring neurocognition, social cognition, functioning and alexithymia. We conducted linear regressions assessing whether alexithymia predicted functional outcomes in BD. Next, we conducted hierarchical stepwise linear regressions investigating the predictive ability of neurocognition, social cognition and alexithymia on everyday and social functioning in our overall sample. BD and SZ patients were comparable on most demographics and demonstrated higher alexithymia compared to HCs. In BD, alexithymia predicted social functioning only. In the overall sample, difficulty identifying and describing feelings predicted everyday functioning; difficulty describing feelings predicted social functioning. Results suggest that aspects of alexithymia significantly predict functioning among these psychiatric groups, above and beyond the contributions of previously identified factors such as neurocognition and social cognition. Results may aid in developing proper interventions aimed at improving patients' ability to articulate their feelings.

Early temperament and negative reactivity in boys with fragile X syndrome.

BACKGROUND: The phenotype of children and adults with fragile X syndrome (FXS) includes a number of problem behaviours such as inattention, social anxiety and aggressive outbursts. However, very little work has been conducted with young children with FXS less than 5 years of age to examine the developmental pathway of problem behaviours in this population and to determine if later occurring problem behaviours may be rooted in early appearing temperament profiles. METHODS: Parent ratings and laboratory-based behavioural observations of negative reactivity were examined in 25 3-year-old boys with FXS and compared with 64 typically developing boys matched on age. RESULTS: Compared with the typically developing group, boys with FXS were rated by their parents as exhibiting less anger and sadness on the Child Behaviour Questionnaire (CBQ), and they showed less facial sadness on the Laboratory Temperament Assessment Battery (Lab-TAB). No group differences were found on the Lab-TAB measures of distress vocalisations, bodily struggle, and facial anger; and anger peaked in the middle of the arm restraint episode for both groups. For boys with FXS, mental age was moderately positively correlated, and autistic behaviour was moderately negatively correlated, with sadness scores from the CBQ. CONCLUSIONS: Our results show different behavioural profiles in very young children with FXS than reported in older-aged children with FXS which implies that temperamental differences and elevated problem behaviours reported in older-aged children with FXS may not be rooted in early temperament. This information is important to develop the phenotype of early development in FXS to facilitate early identification and treatment.

Social cognition moderates the relationship between neurocognition and community functioning in bipolar disorder.

BACKGROUND: Schizophrenia (SZ) studies suggest that neurocognition predicts functional outcome and that social cognition mediates this relationship. Bipolar disorder (BD) patients also have cognitive, social, and functional impairments but the relationship among these factors in BD is not well established. We assessed whether social cognition modulates the influence of neurocognition on community functioning in BD, as found in SZ. METHODS: 200 BD patients and 49 healthy controls (HC) were administered and compared on a battery of tests assessing neurocognition, social cognition, and community functioning. We conducted a series of regression analyses to investigate potential mediation or moderation of social cognition on the relationship between neurocognition and community functioning. RESULTS: BD patients performed worse on neurocognitive domains of processing speed, attention, verbal learning, and global neurocognition. Also, BD patients performed worse on theory of mind, the social cognition composite score, and community functioning. Neurocognition did not significantly predict functional outcome in our BD sample. However, we found a moderating effect of social cognition: among patients with poor social cognition, better neurocognition was associated with better community functioning, a relationship not seen in BD patients with good social cognition. LIMITATIONS: The study was limited by a relatively small HC group and assessing one subtype of functioning status. CONCLUSIONS: The relationship between neurocognition and community functioning in BD may be dependent on social cognition status, implying the presence of social cognitive heterogeneity. Results may be relevant to choosing proper treatment interventions depending on the patient's social cognitive level.

A pilot study investigating two dose reduction techniques for AP lumbar spine radiography using direct dosimetry and Projection VR.

INTRODUCTION: The purpose of this study was to compare radiation dose measurements generated using a virtual radiography simulation with experimental dosimeter measurements for two radiation dose reduction techniques in digital radiography. METHODS: Entrance Surface Dose (ESD) measurements were generated for an antero-posterior lumbar spine radiograph experimentally using NanoDOT™, single point dosimeters, for two radiographic systems (systems 1 and 2) and using Projection VR™, a virtual radiography simulation (system 3). Two dose reduction methods were tested, application of the 15% kVp rule, or simplified 10 kVp rule, and the exposure maintenance formula. The 15% or 10 kVp rules use a specified increase in kVp and halving of the mAs to reduce patient ESD. The exposure maintenance formula uses the increase in source-to-object distance to reduce ESD. RESULTS: Increasing kVp from 75 to 96 kVp, with the concomitant decrease in mAs, resulted in percent ESD reduction of 59.5% (4.02-1.63 mGy), 60.8% (3.55-1.39 mGy), and 60.3% (6.65-2.64 mGy), for experimental systems 1 and 2, and virtual simulation (system 3), respectively. Increasing the SID (with the appropriate increase in mAs) from 100 to 140 cm reduced ESD by 22.3% 18.8%, and 23.5%, for experimental systems 1 and 2, and virtual simulation (system 3), respectively. CONCLUSION: Percent dose reduction measurements were similar between the experimental and virtual measurement systems investigated. For the dose reduction practices tested, Projection VR™ provides a realistic alternate of percent dose reduction to direct dosimetry.

A cost-effectiveness analysis of heroin detoxification methods in the Australian National Evaluation of Pharmacotherapies for Opioid Dependence (NEPOD).

This economic evaluation was part of the Australian National Evaluation of Pharmacotherapies for Opioid Dependence (NEPOD) project. Data from four trials of heroin detoxification methods, involving 365 participants, were pooled to enable a comprehensive comparison of the cost-effectiveness of five inpatient and outpatient detoxification methods. This study took the perspective of the treatment provider in assessing resource use and costs. Two short-term outcome measures were used-achievement of an initial 7-day period of abstinence, and entry into ongoing post-detoxification treatment. The mean costs of the various detoxification methods ranged widely, from AUD 491 dollars(buprenorphine-based outpatient); to AUD 605 dollars for conventional outpatient; AUD 1404 dollars for conventional inpatient; AUD 1990 dollars for rapid detoxification under sedation; and to AUD 2689 dollars for anaesthesia per episode. An incremental cost-effectiveness analysis was carried out using conventional outpatient detoxification as the base comparator. The buprenorphine-based outpatient detoxification method was found to be the most cost-effective method overall, and rapid opioid detoxification under sedation was the most cost-effective inpatient method.

Isolated cardiac myocytes. A new cellular model for studying insulin modulation of monosaccharide transport.

The usefulness of isolated Ca2+-tolerant myocytes as a cellular model system for investigating modulation of monosaccharide transport by insulin was investigated. We have found that the isolation technique described by Haworth et al. (Haworth, R.A., Hunter, D.R. and Berkoff, H.A. (1980) J. Mol. Cell. Cardiol. 12, 715-724), with some minor modifications, consistently gave the highest yield of quiescent, rod-shaped myocytes which maintained their integrity in the presence of 2 mM calcium. Using 3-0-methylglucose, a non-metabolized sugar, transport was shown to possess saturability, substrate stereospecificity, competition and countertransport; all of which have been thoroughly established for D-glucose transport in other systems. The apparent Km of transport ranged from 2.3 to 3.5 mM. Insulin (10 nM) caused a small but significant increase in Km and a 2-3-fold increase in Vmax. These results suggest that this myocyte preparation will provide a useful model for studying the transport-related effects of insulin as well as current hypotheses regarding the mechanism of insulin modulation of transport at the cellular level.

Interactions of insulin, catecholamines and adenosine in the regulation of glucose transport in isolated rat cardiac myocytes.

The regulation of the glucose transport system by catecholamines and insulin has been studied in isolated rat cardiomyocytes. In the basal state, 1-isoproterenol exhibited a biphasic concentration-dependent regulation of 3-O-methylglucose transport. At low concentrations (less than 10 nM), isoproterenol induced a maximal inhibition of 65-70% of the basal rates, while at higher concentrations (greater than 10 nM) a 25-70% stimulation of transport was observed. In the presence of adenosine deaminase, the inhibition of isoproterenol at low doses was attenuated. No effect of adenosine deaminase was observed on the stimulation of transport at high doses of isoproterenol. The inhibitory effect of isoproterenol returned when N6-phenylisopropyladenosine (a non-metabolizable analog of adenosine) was included along with adenosine deaminase. Dibutyryl cAMP and forskolin both inhibited basal transport rates. In the presence of maximally stimulating concentrations of insulin, cardiomyocyte 3-O-methylglucose transport was generally elevated 200-300% above basal levels. In the presence of isoproterenol, insulin stimulation was inhibited at both high and low concentrations of catecholamine, with maximum inhibition occurring at the lowest concentrations tested. When cells were incubated with both adenosine deaminase and isoproterenol, the inhibition of the insulin response was greater at all concentrations of catecholamine and was almost completely blocked at isoproterenol concentrations of 10 nM or less. Dibutyryl cAMP inhibited the insulin response to within 10% of basal transport levels, while forskolin completely inhibited all transport activity in the presence of insulin. These results suggest that catecholamines regulate basal and insulin-stimulated glucose transport via both cAMP-dependent and cAMP-independent mechanisms and that this regulation is modulated in the presence of extracellular adenosine.

Localization of the binding domain of the inhibitory ligand forskolin in the glucose transporter GLUT-4 by photolabeling, proteolytic cleavage and a site-specific antiserum.

The binding domain of forskolin in the adipocyte/muscle-type glucose transporter (GLUT-4) was localized with the aid of the photoreactive derivative, [125I]IAPS-forskolin (3-[125I]iodo-4-azidophenethylamido-7-O-succinyldeacetyl-forskolin). Plasma membranes from insulin-treated rat adipocytes containing predominantly the GLUT-4 isoform were irradiated with UV light in the presence of [125I]IAPS-forskolin. The covalently labeled glucose transporters were isolated by immunoprecipitation with specific antiserum and partially digested with trypsin and elastase. The fragments were separated by gel electrophoresis, transferred on to nitrocellulose membranes, and identified by direct autoradiography and by immunoassay with antiserum against a peptide sequence corresponding to the C-terminus of GLUT-4. Digestion with a high-purity grade trypsin generated two photolabeled fragments with apparent molecular weights of 21 and 16 kDa. Since the antiserum detected two fragments with identical electrophoretic mobility, both labeled fragments appeared to contain the intact C-terminus of GLUT-4. In contrast, digestion with elastase generated only one photolabeled fragment with intact C-terminus at 21 kDa, and a smaller unlabeled fragment with intact C-terminus at 15 kDa. A less pure trypsin preparation generated two labeled (21 and 17 kDa) and one unlabeled (15 kDa) fragment with intact C-terminus. These data suggest that the site of covalent binding of IAPS-forskolin in the GLUT-4 is located within a region of 1-6 kDa defined by the difference between the unlabeled C-terminal fragment (15 kDa) and the labeled fragments (21, 17 and 16 kDa). Based on a tentative allocation of the fragments to the sequence of the GLUT-4, it is suggested that the covalent binding site of IAPS-forskolin is located between the membrane spanning helices 7-9, possibly in the proximity of helix 9.

Glucose transport activity and ligand binding (cytochalasin B, IAPS-forskolin) of chimeric constructs of GLUT2 and GLUT4 expressed in COS-7-cells.

Chimeric constructs of glucose transporters GLUT2 and GLUT4 were transiently expressed in COS-7 cells in order to determine regions of the proteins responsible for their differences in activity and ligand binding. Exchange of the C-terminal tail (aa 479-509) of GLUT4 failed to affect glucose transport activity assayed at 1 mM glucose or ligand binding (cytochalasin B, IAPS-forskolin). In contrast, exchange of the C-terminal half of GLUT4 (aa 222-509) for that of GLUT2 markedly reduced ligand binding (Kd of cytochalasin B binding 1.88 +/- 0.2 microM vs. 0.21 +/- 0.06 in the wild-type GLUT4), and moderately (25%) reduced glucose transport activity. These data support the conclusion that the domains determining differences in ligand binding between GLUT4 and GLUT2 are located in the C-terminal half of the glucose transporters.

Stimulation of glucose transport in rat cardiac myocytes by guanosine 3',5'-monophosphate.

Glucose transport in isolated rat cardiomyocytes is stimulated by insulin, catecholamines, and anoxia approximately 2- to 3-fold over basal rates. The molecular mechanisms controlling these responses are unknown. In our search for possible cellular mediators of glucose transport stimulation, we examined the effects of a number of nucleotides on 3-O-methylglucose transport in heart cells. The nucleotides and/or permeable analogs (monosuccinyl, 8-bromo, and dibutyryl derivatives) included cUMP, cIMP, cCMP, cAMP, and cGMP at concentrations ranging from 10 nM to 1 mM. Of all the nucleotides tested only cGMP analogs induced a significant stimulation of transport at concentrations as low as 100 nM. This effect was observed in both the 8-bromo- and dibutyryl derivatives and with 1 mM cGMP itself. The effect was concentration dependent for both analogs and produced a maximal response equivalent to that of 100 nM insulin. This insulinomimetic effect of cGMP was examined in more detail in order to evaluate its role as a potential mediator of this response. Agents that are known to stimulate guanylate cyclase in the heart produced a clear stimulation of transport when added to cardiomyocytes. These include insulin, aminophylline, histamine, beta-estradiol, and biotin-nitrophenyl ester. Methylene blue, an inhibitor of guanylate cyclase, blocked the insulin response when added to cells before insulin, but was ineffective when added after insulin. In addition, agents that raise intracellular cGMP levels by inhibiting cyclic nucleotide phosphodiesterases were also examined for effects on glucose transport. Out of several phosphodiesterase inhibitors tested, only Zaprinast (which selectively increases cGMP in heart) stimulated transport in a concentration-dependent manner to within 80% of the maximal insulin effect. These results are consistent with the notion that cGMP may be involved in glucose transport stimulation.