α-Melanocyte stimulating hormone promotes muscle glucose uptake via melanocortin 5 receptors

OBJECTIVE: Central melanocortin pathways are well-established regulators of energy balance. However, scant data exist about the role of systemic melanocortin peptides. We set out to determine if peripheral α-melanocyte stimulating hormone (α-MSH) plays a role in glucose homeostasis and tested the hypothesis that the pituitary is able to sense a physiological increase in circulating glucose and responds by secreting α-MSH.

METHODS: We established glucose-stimulated α-MSH secretion using humans, non-human primates, and mouse models. Continuous α-MSH infusions were performed during glucose tolerance tests and hyperinsulinemic-euglycemic clamps to evaluate the systemic effect of α-MSH in glucose regulation. Complementary ex vivo and in vitro techniques were employed to delineate the direct action of α-MSH via the melanocortin 5 receptor (MC5R)-PKA axis in skeletal muscles. Combined treatment of non-selective/selective phosphodiesterase inhibitor and α-MSH was adopted to restore glucose tolerance in obese mice.

RESULTS: Here we demonstrate that pituitary secretion of α-MSH is increased by glucose. Peripheral α-MSH increases temperature in skeletal muscles, acts directly on soleus and gastrocnemius muscles to significantly increase glucose uptake, and enhances whole-body glucose clearance via the activation of muscle MC5R and protein kinase A. These actions are absent in obese mice, accompanied by a blunting of α-MSH-induced cAMP levels in skeletal muscles of obese mice. Both selective and non-selective phosphodiesterase inhibition restores α-MSH induced skeletal muscle glucose uptake and improves glucose disposal in obese mice.

CONCLUSION: These data describe a novel endocrine circuit that modulates glucose homeostasis by pituitary α-MSH, which increases muscle glucose uptake and thermogenesis through the activation of a MC5R-PKA-pathway, which is disrupted in obesity.

Dissecting isoform selectivity of P13K inhibitors: the role of non-conserved residues in the catalytic pocket

The last few years have seen the identification of numerous small molecules that selectively inhibit specific class I isoforms of PI3K (phosphoinositide 3-kinase), yet little has been revealed about the molecular basis for the observed selectivities. Using site-directed mutagenesis, we have investigated one of the areas postulated as being critical to the observed selectivity. The residues Thr⁸⁸⁶ and Lys⁸⁹⁰ of the PI3Kγ isoform project towards the ATP-binding pocket at the entrance to the catalytic site, but are not conserved. We have made reciprocal mutations between those residues in the β isoform (Glu⁸⁵⁸ and Asp⁸⁶²) and those in the α isoform (His⁸⁵⁵ and Gln⁸⁵⁹) and evaluated the potency of a range of reported PI3K inhibitors. The results show that the potencies of β-selective inhibitors TGX221 and TGX286 are unaffected by this change. In contrast, close analogues of these compounds, particularly the α-isoform-selective compound (III), are markedly influenced by the point mutations. The collected data suggests two distinct binding poses for these inhibitor classes, one of which is associated with potent PI3Kβ activity and is not associated with the mutated residues, and a second that, in accord with earlier hypotheses, does involve this pair of non-conserved amino acids at the catalytic site entrance and contributes to the α-isoform-selectivity of the compounds studied.

Measuring non-economic well-being achievement

Income per capita and most widely reported, non- or non-exclusively income based human well-being indicators are highly correlated among countries. Yet many countries exhibit higher achievement in the latter than predicted by the former. The reverse is true for many other countries. This paper commences by extracting the inter-country variation in a composite of various widely-reported, non-income-based well-being indices not accounted for by variations in income pre capita. This extraction is interpreted inter alia as a measure of non-economic well-being. The paper then looks at correlations between this extraction and a number of new or less widely-used well-being measures, in an attempt to find the measure that best captures these achievements. A number of indicators are examined, including measures of poverty, inequality, health status, education status, gender bias, empowerment, governance and subjective well-being.

Non-economic well-being achievement in Pacific Asia

It is well known that income per capita and most widely reported non-economic well-being achievement measures are highly correlated among countries. Yet many countries exhibit higher achievement in the latter than predicted by the former. The reverse is true for many other countries. This paper commences by extracting the inter-country variation in a composite of three widely reported educational and health status indicators not accounted for by variations in income per capita. This extraction is interpreted inter alia as a measure of non-economic well-being. Using data for a sample of Pacific Asian countries, the paper then looks at correlations between this extraction and a number of new or less widely-used well-being measures, in an attempt to find the measure that best captures these achievements.

Insulin-regulated aminopeptidase : analysis of peptide substrate and inhibitor binding to the catalytic domain

Peptide inhibitors of insulin-regulated aminopeptidase (IRAP) accelerate spatial learning and facilitate memory retention and retrieval by binding competitively to the catalytic site of the enzyme and inhibiting its catalytic activity. IRAP belongs to the M1 family of Zn2+-dependent aminopeptidases characterized by a catalytic domain that contains two conserved motifs, the HEXXH(X)18E Zn2+-binding motif and the GXMEN exopeptidase motif. To elucidate the role of GXMEN in binding peptide substrates and competitive inhibitors, site-directed mutagenesis was performed on the motif. Non-conserved mutations of residues G428, A429 and N432 resulted in mutant enzymes with altered catalytic activity, as well as divergent changes in kinetic properties towards the synthetic substrate leucine β-naphthalamide. The affinities of the IRAP inhibitors angiotensin IV, Nle1-angiotensin IV, and LVV-hemorphin-7 were selectively decreased. Substrate degradation studies using the in vitro substrates vasopressin and Leu-enkephalin showed that replacement of G428 by either D, E or Q selectively abolished the catalysis of Leu-enkephalin, while [A429G]IRAP and [N432A]IRAP mutants were incapable of cleaving both substrates. These mutational studies indicate that G428, A429 and N432 are important for binding of both peptide substrates and inhibitors, and confirm previous results demonstrating that peptide IRAP inhibitors competitively bind to its catalytic site.

Anisotropic hardening model based on non-associated flow rule and combined nonlinear kinematic hardening for sheet materials

 A material model for more effective analysis of plastic deformation of sheet materials is presented in this paper. The model is capable of considering the following aspects of plastic deformation behavior of sheet materials: the anisotropy in yielding stresses in different directions by using a quadratic yield function (based on Hill’s 1948 model and stress ratios), the anisotropy in work hardening by introducing non-constant flow stress hardening in different directions, the anisotropy in plastic strains in different directions by using a quadratic plastic potential function and non-associated flow rule (based on Hill’s 1948 model and plastic strain ratios, r-values), and finally some of the cyclic hardening phenomena such as Bauschinger’s effect and transient behavior for reverse loading by using a coupled nonlinear kinematic hardening (so-called Armstrong-Frederick-Chaboche model). Basic fundamentals of the plasticity of the model are presented in a general framework. Then, the model adjustment procedure is derived for the plasticity formulations. Also, a generic numerical stress integration procedure is developed based on backward-Euler method (so-called multistage return mapping algorithm). Different aspects of the model are verified for DP600 steel sheet. Results show that the new model is able to predict the sheet material behavior in both anisotropic hardening and cyclic hardening regimes more accurately. By featuring the above-mentioned facts in the presented constitutive model, it is expected that more accurate results can be obtained by implementing this model in computational simulations of sheet material forming processes. For instance, more precise results of springback prediction of the parts formed from highly anisotropic hardened materials or that of determining the forming limit diagrams is highly expected by using the developed material model.

A non-associated plasticity model with anisotropic and nonlinear kinematic hardening for simulation of sheet metal forming

A material model for more thorough analysis of plastic deformation of sheet materials is presented in this paper. This model considers the following aspects of plastic deformation behavior of sheet materials: (1) the anisotropy in yield stresses and in work hardening by using Hill's 1948 quadratic yield function and non-constant stress ratios which leads to different flow stress hardening in different directions, (2) the anisotropy in plastic strains by using a quadratic plastic potential function and non-associated flow rule, also based on Hill's 1948 model and r-values, and (3) the cyclic hardening phenomena such as the Bauschinger effect, permanent softening and transient behavior for reverse loading by using a coupled nonlinear kinematic hardening model. Plasticity fundamentals of the model were derived in a general framework and the model calibration procedure was presented for the plasticity formulations. Also, a generic numerical stress integration procedure was developed based on backward-Euler method, so-called multi-stage return mapping algorithm. The model was implemented in the framework of the finite element method to evaluate the simulation results of sheet metal forming processes. Different aspects of the model were verified for two sheet metals, namely DP600 steel and AA6022 aluminum alloy. Results show that the new model is able to accurately predict the sheet material behavior for both anisotropic hardening and cyclic hardening conditions. The drawing of channel sections and the subsequent springback were also simulated with this model for different drawbead configurations. Simulation results show that the current non-associated anisotropic hardening model is able to accurately predict the sidewall curl in the drawn channel sections.

A non-destructive test to assess the axial heterogeneity of in situ modified monoliths for HPLC

This paper describes a non-destructive "peak parking" protocol in order to assess the axial heterogeneity of an in situ modified monolithic column for high performance liquid chromatography; a "gradient stationary phase" was designed whereby the ligand density decreases along the length of the rod in the "forward flow" configuration. The results of multi-location peak parking demonstrated a consistent increase in peak variance from the 1 cm position of the column to the 9 cm location. This increase in band broadening supported the theory of a decreasing ligand density along the length of this gradient column. This is consistent with efficiency measurements performed in both the forward and reverse flow directions, with an improved efficiency (15% increase in N m^-1) in the reverse direction. These results are consistent with theoretical investigations into stationary phase gradients.

Cancer risk among people with type 1 and type 2 diabetes: disentangling true associations, detection bias, and reverse causation

 OBJECTIVE: Evidence indicates an increased risk of certain cancers among people with type 2 diabetes. Evidence for rarer cancers and for type 1 diabetes is limited. We explored the excess risk of site-specific cancer incidence and mortality among people with type 1 and type 2 diabetes, compared with the general Australian population. RESEARCH DESIGN AND METHODS: Registrants of a national diabetes registry (953,382) between 1997 and 2008 were linked to national death and cancer registries. Standardized incidence and mortality ratios (SIRs/SMRs) are reported. RESULTS: For type 1 diabetes, significant elevated SIRs were observed for pancreas, liver, esophagus, colon and rectum (females only [F]), stomach (F), thyroid (F), brain (F), lung (F), endometrium, and ovary, and decreased SIRs were observed for prostate in males. Significantly increased SMRs were observed for pancreas, liver, and kidney (males only), non-Hodgkin's lymphoma, brain (F), and endometrium. For type 2 diabetes, significant SIRs were observed for almost all site-specific cancers, with highest SIRs observed for liver and pancreas, and decreased risks for prostate and melanoma. Significant SMRs were observed for liver, pancreas, kidney, Hodgkin's lymphoma, gallbladder (F), stomach (F), and non-Hodgkin's lymphoma (F). Cancer risk was significantly elevated throughout follow-up time but was higher in the first 3 months postregistration, suggesting the presence of detection bias and/or reverse causation. CONCLUSIONS: Type 1 and type 2 diabetes are associated with an excess risk of incidence and mortality for overall and a number of site-specific cancers, and this is only partially explained by bias. We suggest that screening for cancers in diabetic patients is important.

Meropenem versus piperacillin-tazobactam for definitive treatment of bloodstream infections due to ceftriaxone non-susceptible Escherichia coli and Klebsiella spp (the MERINO trial): study protocol for a randomised controlled trial

BACKGROUND: Gram-negative bacteria such as Escherichia coli or Klebsiella spp. frequently cause bloodstream infections. There has been a worldwide increase in resistance in these species to antibiotics such as third generation cephalosporins, largely driven by the acquisition of extended-spectrum beta-lactamase or plasmid-mediated AmpC enzymes. Carbapenems have been considered the most effective therapy for serious infections caused by such resistant bacteria; however, increased use creates selection pressure for carbapenem resistance, an emerging threat arising predominantly from the dissemination of genes encoding carbapenemases. Recent retrospective data suggest that beta-lactam/beta-lactamase inhibitor combinations, such as piperacillin-tazobactam, may be non-inferior to carbapenems for the treatment of bloodstream infection caused by extended-spectrum beta-lactamase-producers, if susceptible in vitro. This study aims to test this hypothesis in an effort to define carbapenem-sparing alternatives for these infections.

METHODS/DESIGN: The study will use a multicentre randomised controlled open-label non-inferiority trial design comparing two treatments, meropenem (standard arm) and piperacillin-tazobactam (carbapenem-sparing arm) in adult patients with bacteraemia caused by E. coli or Klebsiella spp. demonstrating non-susceptibility to third generation cephalosporins. Recruitment is planned to occur in sites across three countries (Australia, New Zealand and Singapore). A total sample size of 454 patients will be required to achieve 80% power to determine non-inferiority with a margin of 5%. Once randomised, definitive treatment will be for a minimum of 4 days, but up to 14 days with total duration determined by treating clinicians. Data describing demographic information, antibiotic use, co-morbid conditions, illness severity, source of infection and other risk factors will be collected. Vital signs, white cell count, use of vasopressors and days to bacteraemia clearance will be recorded up to day 7. The primary outcome measure will be mortality at 30 days, with secondary outcomes including days to clinical and microbiological resolution, microbiological failure or relapse, isolation of a multi-resistant organism or Clostridium difficile infection.

Dangerous practices : the practicum experiences of non-Indigenous pre-service teachers in remote communities

This paper seeks to explore the risks of providing preserviceteachers with professional experiences in remote communities.In particular this paper focuses on the risks associated with this kindof professional experience. Twelve pre-service teachers wereinterviewed whilst on a three-week practicum around Katherine andin Maningrida in the Northern Territory during 2012. The dangersoutlined in this paper relate to the way their experiences continued tobe mediated by stereotypes and perpetuating colonial practices. Thepre-service teachers’ limited understandings of Indigenousknowledges and languages are discussed before exploring the vexedissue of reverse culture shock that some of the participants identifiedwhen they returned home. The paper concludes by exploring thenotion of ‘allies’ as a way to negotiate the problematic nature of thiswork.

Mitigation of N₂O emissions from surface-irrigated cropping systems using water management and the nitrification inhibitor DMPP

Soils under irrigated agriculture are a significant source of nitrous oxide (N2O) owing to high inputs of nitrogen (N) fertiliser and water. This study investigated the potential for N2O mitigation by manipulating the soil moisture deficit through irrigation scheduling in combination with, and in comparison to, using the nitrification inhibitor, 3,4-dimethylpyrazole phosphate (DMPP). Lysimeter cores planted with wheat were fitted with automated chambers for continuous measurements of N2O fluxes. Treatments included conventional irrigation (CONV), reduced deficit irrigation (RED), CONV-DMPP and RED-DMPP. The total seasonal volume of irrigation water applied was constant for all treatments but the timing and quantity in individual irrigation applications varied among treatments. 15N-labelled urea was used to track the source of N2O emissions and plant N uptake. The majority of N2O emissions occurred immediately after irrigations began on 1 September 2014. Applying RED and DMPP individually slightly decreased N2O emissions but when applied in combination (RED-DMPP) the greatest reductions in N2O emissions were observed. There was no effect of treatments on plant N uptake, 15N recovery or yield possibly because the system was not N limited. Half of the plant N and 53% to 87% of N2O was derived from non-fertiliser sources in soil, highlighting the opportunity to further exploit this valuable N pool.