Intra-islet glucagon signalling regulates beta-cell connectivity, first-phase insulin secretion and glucose homoeostasis.

OBJECTIVE: Type 2 diabetes (T2D) is characterised by the loss of first-phase insulin secretion. We studied mice with ß-cell selective loss of the glucagon receptor (Gcgrfl/fl X Ins-1Cre), to investigate the role of intra-islet glucagon receptor (GCGR) signalling on pan-islet [Ca2+]I activity and insulin secretion. METHODS: Metabolic profiling was conducted on Gcgrß-cell-/- and littermate controls. Crossing with GCaMP6f (STOP flox) animals further allowed for ß-cell specific expression of a fluorescent calcium indicator. These islets were functionally imaged in vitro and in vivo. Wild-type mice were transplanted with islets expressing GCaMP6f in ß-cells into the anterior eye chamber and placed on a high fat diet. Part of the cohort received a glucagon analogue (GCG-analogue) for 40 days and the control group were fed to achieve weight matching. Calcium imaging was performed regularly during the development of hyperglycaemia and in response to GCG-analogue treatment. RESULTS: Gcgrß-cell-/- mice exhibited higher glucose levels following intraperitoneal glucose challenge (control 12.7 mmol/L ± 0.6 vs. Gcgrß-cell-/- 15.4 mmol/L ± 0.0 at 15 min, p = 0.002); fasting glycaemia was not different to controls. In vitro, Gcgrß-cell-/- islets showed profound loss of pan-islet [Ca2+]I waves in response to glucose which was only partially rescued in vivo. Diet induced obesity and hyperglycaemia also resulted in a loss of co-ordinated [Ca2+]I waves in transplanted islets. This was reversed with GCG-analogue treatment, independently of weight-loss (n = 8). CONCLUSION: These data provide novel evidence for the role of intra-islet GCGR signalling in sustaining synchronised [Ca2+]I waves and support a possible therapeutic role for glucagonergic agents to restore the insulin secretory capacity lost in T2D.

An innovative analysis of nasolabial dynamics of surgically managed adult patients with unilateral cleft lip and palate using 3D facial motion capture.

AIM: To compare dynamic nasolabial movement between end-of-treatment cleft and a matched non-cleft group in adult patients. MATERIALS AND METHODS: Thirteen treated adult participants with unilateral cleft lip and palate had images taken using a facial motion capture system performing a maximum smile. Seventeen landmarks were automatically tracked. For each landmark pair, on either side of the midline, changes in the x, y, and z directions were used to analyze the magnitude of displacement and path of motion. An asymmetry score was developed at rest, mid-smile, and maximum smile to assess the shape of the mouth and/or nose. RESULTS: At maximum smile, displacement of right and left cheilion was clinically and statistically (p < 0.05) less in the cleft group. The lip asymmetry score was greater (p < 0.05) at each time point in the cleft group using the clinical midline. Using Procrustes superimposition, the differences were significant (p < 0.05) only at rest and mid-smile. The alar bases were displaced significantly less (p < 0.05) in the z direction in the cleft group. The asymmetry score of the alar base was significantly higher using the clinical midline than using Procrustes superimposition in patients with cleft conditions (p < 0.001). In the cleft group, at maximum smile, the right and left cristae philter moved significantly less (p < 0.05) in the x and z directions. CONCLUSIONS: There was an increase in asymmetry score of the corners of the mouth and alar bases from rest to maximum smile. The lips were similar in shape but oriented differently in the faces of patients with cleft conditions than in individuals without those conditions.

Incidence of Deep Venous Thrombosis and Sickle Cell Disease in Patients Undergoing Spinal Surgery in South Gujarat, India: A Prospective Observational Study.

Introduction: Our objective of this study was to assess the incidence of Deep Venous Thrombosis in patients including those with sickle cell disease who underwent spine surgery, and also to determine the association of Sickle Cell Disease as a clinical predictor for Deep Venous Thrombosis in spinal surgery patients. Materials and methods: All patients who underwent spinal surgery from January 2016 to October 2016 were included in this study. Detailed history, demographic data, physical findings, pre-operative haematological and radiological investigations were documented. All the patients underwent daily clinical evaluation for clinical signs of Deep Venous Thrombosis and also underwent a post-operative venous Doppler and D-dimer test. Results: Seventy-nine consecutive patients were included in the study with the mean age of 41 years. All patients had normal venous Doppler pre-operatively. A total of 2.5% patients had deep vein thrombosis in bilateral lower limbs while 2 patients (2.5%) had evidence of venous stasis but no thrombosis on Doppler ultrasound done post-operatively. Nine patients (11.4%) were sickle cell positive from which 4 patients showed evidence of Deep Venous Thrombosis or Venous Stasis. D-dimer was positive in 5 (8.3%) patients which included 4 patients with Sickle Cell Disease. Conclusion: This study concludes that Sickle Cell Disease is a risk factor for developing Deep Venous Thrombosis in patients undergoing spinal surgery. The study also concludes the effectiveness of mechanical prophylaxis in preventing Deep Venous Thrombosis and recommends pharmacological prophylaxis after assessing the risk profile or positive D-dimer test.

Strengthened capacity of India´s bedaquiline Conditional Access Programme for introducing new drugs and regimens.

BACKGROUND: Addressing TB in India is critical to meeting global targets. With the scale-up of diagnostic networks and the availability of new TB drugs, India had the opportunity to improve the detection and treatment outcomes in drug-resistant TB (DR-TB).OBJECTIVE: To document how the introduction of new drugs and regimens is helping India improve the care of DR-TB patients.DESIGN: In 2016, India´s National TB Programme (NTP) introduced bedaquiline (BDQ) under a Conditional Access Programme (BDQ-CAP) at six sites after providing extensive training and strengthening laboratory testing, pre-treatment evaluation, active drug safety monitoring and management (aDSM) and follow-up systems.RESULTS: An interim analysis reflected earlier and better culture conversion rates: 83% of the 620 patients converted within a median time of 60 days. However, 248 serious adverse events were reported, including 73 deaths (12%) and 100 cardiotoxicity events (16.3%). Encouraged by the evidence of safety and efficacy of BDQ, the NTP took steps to systematically expand its access to cover the entire population by 2018.CONCLUSION: The cautious yet focused approach used to introduce BDQ under BDQ-CAP paved the way for the rapid introduction of delamanid, as well as the shorter treatment regimen and the all-oral regimen for DR-TB.

Overexpression of miR-489 derails mammary hierarchy structure and inhibits HER2/neu-induced tumorigenesis.

Although it has been demonstrated that transformed progenitor cell population can contribute to tumor initiation, factors contributing to this malignant transformation are poorly known. Using in vitro and xenograft-based models, previous studies demonstrated that miR-489 acts as a tumor suppressor miRNA by targeting various oncogenic pathways. It has been demonstrated that miR-489 directly targets HER2 and inhibits the HER2 signaling pathway; however, its role in mammary gland development and HER2-induced tumor initiation hasn't been studied. To dissect the role of miR-489, we sorted different populations of mammary epithelial cells and determined that miR-489 was highly expressed in mammary stem cells. MMTV-miR-489 mice that overexpressed miR-489 in mammary epithelial cells were developed and these mice exhibited an inhibition of mammary gland development in early ages with a specific impact on highly proliferative cells. Double transgenic MMTV-Her2-miR489 mice were then generated to observe how miR-489 overexpression affects HER2-induced tumorigenesis. miR-489 overexpression delayed HER2-induced tumor initiation significantly. Moreover, miR-489 overexpression inhibited tumor growth and lung metastasis. miR-489 overexpression reduced mammary progenitor cell population significantly in preneoplastic mammary glands of MMTV-Her2 mice which showed a putative transformed population in HER2-induced tumorigenesis. The miR-489 overexpression reduced CD49fhiCD61hi populations in tumors that have stem-like properties, and miR-489 overexpression altered the HER2 signaling pathway in mammary tumors. Altogether, these data indicate that the inhibition of HER2-induced tumorigenesis by miR-489 overexpression was due to altering progenitor cell populations while decreasing tumor growth and metastasis via influencing tumor promoting genes DEK and SHP2.

Maturation of the Human Cerebral Cortex During Adolescence: Myelin or Dendritic Arbor?

Previous in vivo studies revealed robust age-related variations in structural properties of the human cerebral cortex during adolescence. Neurobiology underlying these maturational phenomena is largely unknown. Here we employ a virtual-histology approach to gain insights into processes associated with inter-regional variations in cortical microstructure and its maturation, as indexed by magnetization transfer ratio (MTR). Inter-regional variations in MTR correlate with inter-regional variations in expression of genes specific to pyramidal cells (CA1) and ependymal cells; enrichment analyses indicate involvement of these genes in dendritic growth. On the other hand, inter-regional variations in the change of MTR during adolescence correlate with inter-regional profiles of oligodendrocyte-specific gene expression. Complemented by a quantitative hypothetical model of the contribution of surfaces associated with dendritic arbor (1631 m2) and myelin (48 m2), these findings suggest that MTR signals are driven mainly by macromolecules associated with dendritic arbor while maturational changes in the MTR signal are associated with myelination.

Correction: Overexpression of miR-489 derails mammary hierarchy structure and inhibits HER2/neu-induced tumorigenesis.

In the published version of this paper the author A. Awgulewitsch's surname was incorrectly given as Awagulerwitsch instead of Awgulewitsch. This has now been corrected in the HTML version of the paper, the PDF was correct at the time of publication.

Seroprevalence of brucellosis in small ruminants in organized and unorganized sectors of Gujarat state, India.

AIM: The present study aimed to study the seroprevalence of brucellosis in small ruminants of Gujarat state, India, using Rose Bengal Plate test (RBPT) and indirect enzyme-linked immunosorbent assay (iELISA). MATERIALS AND METHODS: A total of 2444 sera samples (675 sheep and 1769 goat) from unorganized sector and 1310 sera samples (861 sheep and 449 goat) from seven organized farms were collected for brucellosis screening. RESULTS: In unorganized sector, 23.70% sheep (160/675) and 15.99% goat (283/1769) were positive by RBPT and 24.44% sheep (165/675) and 17.24% goat (305/1769) by iELISA. The organized sector samples showed higher seroprevalence in goat (7.79 %, 35/449) than sheep (4.06 %, 35/861) by RBPT. Similarly, in iELISA, goat samples showed a higher seroprevalence (9.35%, 42/449) compared to sheep (7.50%, 65/861). The diagnostic sensitivity and specificity of RBPT with ELISA were 88.69% and 99.65%, respectively, and showed a significant difference (p≤0.0001). The Chi-square analysis revealed a significant difference in seroprevalence between sectors (p≤0.01) and species (p≤0.01). CONCLUSION: The seroprevalence of brucellosis in small ruminants of Gujarat was investigated and showed a higher prevalence of brucellosis and warrants the implementation of proper preventive measures.

Revisiting the evolutionary origins of obesity: lazy versus peppy-thrifty genotype hypothesis.

The recent global obesity epidemic is attributed to major societal and environmental changes, such as excessive energy intake and sedentary lifestyle. However, exposure to 'obesogenic' environments does not necessarily result in obesity at the individual level, as 40-75% of body mass index variation in population is attributed to genetic differences. The thrifty genotype theory posits that genetic variants promoting efficient food sequestering and optimal deposition of fat during periods of food abundance were evolutionarily advantageous for the early hunter-gatherer and were positively selected. However, the thrifty genotype is likely too simplistic and fails to provide a justification for the complex distribution of obesity predisposing gene variants and for the broad range of body mass index observed in diverse ethnic groups. This review proposes that gene pleiotropy may better account for the variability in the distribution of obesity susceptibility alleles across modern populations. We outline the lazy-thrifty versus peppy-thrifty genotype hypothesis and detail the body of evidence in the literature in support of this novel concept. Future population genetics and mathematical modelling studies that account for pleiotropy may further improve our understanding of the evolutionary origins of the current obesity epidemic.

A Systematic Review and Meta-Analysis of Baseline Ohip-Edent Scores.

BACKGROUND: OHIP-EDENT is widely used in the literature to assess Oral-Health-Related-Quality-of-Life (OHRQoL) for edentulous patients. However the normal variance and mean of the baseline OHIP scores has not been reported. It would facilitate critical appraisal of studies if we had knowledge of the normal variation and mean of baseline OHIP-EDENT scores. An established figure for baseline OHIP-EDENT, obtained from a meta-analysis, would simplify comparisons of studies and quantify variations in initial OHRQoL of the trial participants. OBJECTIVES: The aim of this study is to quantify a normal baseline value for pre-operative OHIP-EDENT scores by a systematic review and meta-analysis of the available literature. METHODS: A systematic literature review was carried. 83 papers were identified that included OHIP-EDENT values. After screening and eligibility assessment, 7 papers were selected and included in the meta-analysis. RESULTS: A meta-analysis for the 7 papers by a random-effect model yielded a mean baseline OHIP-EDENT score of 28.63 with a 95% Confidence intervals from 21.93 to 35.34. CONCLUSION: A pre-operative baseline OHIP-EDENT has been established by meta-analysis of published papers. This will facilitate the comparison of the initial OHRQoL of one study population to that found elsewhere in the published literature.

Risk factors for biopsy-proven advanced non-alcoholic fatty liver disease in the Veterans Health Administration.

BACKGROUND: With its increasing incidence, nonalcoholic fatty liver disease (NAFLD) is of particular concern in the Veterans Health Administration (VHA). AIMS: To evaluate risk factors for advanced fibrosis in biopsy-proven NAFLD in the VHA, to identify patients at risk for adverse outcomes. METHODS: In randomly selected cases from VHA databases (2005-2015), we performed a retrospective case-control study in adults with biopsy-defined NAFLD or normal liver. RESULTS: Of 2091 patients reviewed, 399 met inclusion criteria. Normal controls (n = 65) had normal liver function. The four NAFLD cohorts included: NAFL steatosis (n = 76), nonalcoholic steatohepatitis (NASH) without fibrosis (n = 68), NAFLD/NASH stage 1-3 fibrosis (n = 82), and NAFLD/NASH cirrhosis (n = 70). NAFLD with hepatocellular carcinoma (HCC) was separately identified (n = 38). Most patients were older White men. NAFLD patients with any fibrosis were on average severely obese (BMI>35 kg/m2 ). Diabetes (54.4%-79.6%) and hypertension (85.8%-100%) were more common in NAFLD with fibrosis or HCC. Across NAFLD, 12.3%-19.5% were enrolled in diet/exercise programs and 0%-2.6% had bariatric surgery. Hispanics exhibited higher rates of NASH (20.6%), while Blacks had low NAFLD rates (1.4%-11.8%), particularly NAFLD cirrhosis and HCC (1.4%-2.6%). Diabetes (OR 11.8, P < .001) and BMI (OR 1.4, P < .001) were the most significant predictors of advanced fibrosis. CONCLUSIONS: In the VHA, diabetes and severe obesity increased risk for advanced fibrosis in NAFLD. Of these patients, only a small proportion (~20%) had enrolled in diet/exercise programs or had bariatric surgery (~2%). These results suggest that providers should focus/tailor interventions to improve outcomes, particularly in those with diabetes and severe obesity.

Effect of season on physiological, biochemical, hormonal, and oxidative stress parameters of indigenous sheep.

AIM: This study was conducted to evaluate the impact of summer and winter season on physiological, biochemical, hormonal, and antioxidant parameters in Indigenous sheep. MATERIALS AND METHODS: The research was carried out during summer and winter season. 8 adult apparently healthy female sheep (aged 2-4 years) of similar physiological status were selected. Daily ambient temperature and relative humidity were recorded to calculate the temperature-humidity index (THI). The THI value of summer and winter season were 82.55 and 59.36, respectively, which indicate extreme hot condition during summer season and extreme cold condition during winter season. Physiological parameters were recorded daily during the experimental periods. Blood samples were collected at weekly interval and analyzed for biochemical, hormonal, and antioxidant parameters. The results were analyzed using completely randomized design. RESULTS: From data obtained in this study, we found that higher THI during summer have significant effect over various physiological, biochemical, hormonal, and enzymatic indices of indigenous sheep. The physiological response such as rectal temperature, respiration rate (RR), pulse rate (PR), and skin temperature (ST) was increased significantly. We also found a significant increase in some biochemical parameters such as blood urea nitrogen (BUN), uric acid, creatinine (Cr), alanine transaminase (ALT), aspartate transaminase (AST), sodium (Na), and potassium (K). The level of cortisol hormone and superoxide dismutase (SOD), glutathione peroxidase (GPx), and lipid peroxidase (LPO) antioxidants increased significantly during summer. Whereas, some parameters such as glucose, cholesterol, calcium (Ca), inorganic phosphorus (IP), triiodothyronine (T3), and thyroxine (T4) were decreased significantly during summer season. CONCLUSION: It was concluded that the THI is a sensitive indicator of heat stress and is impacted by ambient temperature more than the relative humidity in Indigenous sheep. Higher THI is associated with significant increase in RT, RR, PR, ST, BUN, uric acid, Cr, ALT, AST, Na, K, cortisol, SOD, GPx, and LPO and with a significant decrease in glucose, cholesterol, Ca, IP, T3 and T4.

Population Pharmacokinetics of Selumetinib and Its Metabolite N-desmethyl-selumetinib in Adult Patients With Advanced Solid Tumors and Children With Low-Grade Gliomas.

Selumetinib (AZD6244, ARRY-142886), a mitogen activated protein kinases (MEK1 and 2) inhibitor, has been granted orphan drug designation for differentiated thyroid cancer. The primary aim of this analysis was to characterize the population pharmacokinetics of selumetinib and its active metabolite N-desmethyl-selumetinib in patients with cancer. Concentration-time data from adult and pediatric clinical trials were pooled to develop a population pharmacokinetic model using a sequential approach where selumetinib and N-desmethyl-selumetinib data were modeled separately. A sequential zero- and first-order absorption with lag time with a two-compartment model for selumetinib and a two-compartment model for N-desmethyl-selumetinib best described the concentration-time data. Intrapatient variability in absorption was higher than interpatient variability. The apparent drug clearance (CL/F) from the central compartment was 13.5 L/hr (RSE 4.9%). Significant covariates for CL/F were age, alanine aminotransferase, and body surface area. This study confirms that flat dosing is appropriate in adults, whereas body-surface area based dosing should be used in pediatric patients.

Changes in Weight and Glucose Can Protect Against Progression in Early Diabetes Independent of Improvements in ß-Cell Function.

CONTEXT: Evidence-based strategies to prevent progression of dysglycemia in newly diagnosed type 2 diabetes are needed. OBJECTIVE: To undertake a secondary analysis of the Early Diabetes Intervention Program (EDIP) in order to understand the features that were protective against worsening glycemia. DESIGN: EDIP was a randomized, placebo-controlled trial. SETTING: Two university diabetes centers. PATIENTS: A total of 219 overweight individuals with fasting glucose < 7.8 mmol/L and 2-hour oral glucose tolerance test (OGTT) glucose > 11.1 mmol/L. INTERVENTIONS: Acarbose versus placebo, on a background of dietary recommendations, with quarterly visits to assess glycemia and intervention adherence for up to 5 years. MAIN OUTCOME MEASURES: Progression of fasting glucose ≥ 7.8 mmol/L on two consecutive quarterly visits. Cox proportional hazards modeling and ANOVA were performed to evaluate determinants of progression. RESULTS: Progression-free status was associated with reductions in weight, fasting glucose, 2-hour OGTT glucose, and increases in the high-density lipoprotein/triglyceride ratio. The reduction in fasting glucose was the only effect that remained significantly associated with progression-free status in multivariable Cox modeling. The reduction in fasting glucose was in turn primarily associated with reductions in weight and in 2-hour OGTT glucose. Acarbose treatment did not explain these changes. CONCLUSIONS: In early diabetes, reductions in glucose, driven by reductions in weight, can delay progressive metabolic worsening. These observations underscore the importance of lifestyle management including weight loss as a tool to mitigate worsening of glycemia in newly diagnosed diabetes.

Fundamentals of cutting.

The process of cutting is analysed in fracture mechanics terms with a view to quantifying the various parameters involved. The model used is that of orthogonal cutting with a wedge removing a layer of material or chip. The behaviour of the chip is governed by its thickness and for large radii of curvature the chip is elastic and smooth cutting occurs. For smaller thicknesses, there is a transition, first to plastic bending and then to plastic shear for small thicknesses and smooth chips are formed. The governing parameters are tool geometry, which is principally the wedge angle, and the material properties of elastic modulus, yield stress and fracture toughness. Friction can also be important. It is demonstrated that the cutting process may be quantified via these parameters, which could be useful in the study of cutting in biology.

Fracture investigation in starch-based foods.

The study of oral processing and specifically cutting of the food piece during mastication can lead towards optimization of products for humans or animals. Food materials are complex biocomposites with a highly nonlinear constitutive response. Their fracture properties have not been largely investigated, while the need for models capable of predicting food breakdown increases. In this study, the blade cutting and the essential work of fracture (EWF) methodologies assessed the fracture behaviour of starch-based pet food. Tensile tests revealed rate-dependent stiffness and stress softening effects, attributed to viscoplasticity and micro-cracking, respectively. Cutting data were collected for 5, 10 and 30 mm s(-1) sample feed rates, whereas the EWF tests were conducted at 1.7, 3.3 and 8.3 mm s(-1) crosshead speeds corresponding to average crack speeds of 4, 7 and 15 mm s(-1), respectively. A reasonable agreement was achieved between cutting and EWF, reporting 1.26, 1.78, 1.76 kJ m(-2) and 1.52, 1.37, 1.45 kJ m(-2) values, respectively, for the corresponding crack speeds. These toughness data were used in a novel numerical model simulating the 'first' bite mastication process. A viscoplastic material model is adopted for the food piece, combined with a damage law that enabled predicting fracture patterns in the product.

Translational Pharmacokinetic-Pharmacodynamic Modeling and Simulation: Optimizing 5-Fluorouracil Dosing in Children With Pediatric Ependymoma.

We previously investigated novel therapies for pediatric ependymoma and found 5-fluorouracil (5-FU) i.v. bolus increased survival in a representative mouse model. However, without a quantitative framework to derive clinical dosing recommendations, we devised a translational pharmacokinetic-pharmacodynamic (PK-PD) modeling and simulation approach. Results from our preclinical PK-PD model suggested tumor concentrations exceeded the 1-hour target exposure (in vitro IC90), leading to tumor growth delay and increased survival. Using an adult population PK model, we scaled our preclinical PK-PD model to children. To select a 5-FU dosage for our clinical trial in children with ependymoma, we simulated various 5-FU dosages for tumor exposures and tumor growth inhibition, as well as considering tolerability to bolus 5-FU administration. We developed a pediatric population PK model of bolus 5-FU and simulated tumor exposures for our patients. Simulations for tumor concentrations indicated that all patients would be above the 1-hour target exposure for antitumor effect.