Using rapid indicators for Enterococcus to assess the risk of illness after exposure to urban runoff contaminated marine water

Background: Traditional fecal indicator bacteria (FIB) measurement is too slow (>18 h) for timely swimmer warnings. Objectives: Assess relationship of rapid indicator methods (qPCR) to illness at a marine beach impacted by urban runoff. Methods: We measured baseline and two-week health in 9525 individuals visiting Doheny Beach 2007-08. Illness rates were compared (swimmers vs. non-swimmers). FIB measured by traditional (Enterococcus spp. by EPA Method 1600 or Enterolert™, fecal coliforms, total coliforms) and three rapid qPCR assays for Enterococcus spp. (Taqman, Scorpion-1, Scorpion-2) were compared to health. Primary bacterial source was a creek flowing untreated into ocean; the creek did not reach the ocean when a sand berm formed. This provided a natural experiment for examining FIB-health relationships under varying conditions. Results: We observed significant increases in diarrhea (OR 1.90, 95% CI 1.29-2.80 for swallowing water) and other outcomes in swimmers compared to non-swimmers. Exposure (body immersion, head immersion, swallowed water) was associated with increasing risk of gastrointestinal illness (GI). Daily GI incidence patterns were different: swimmers (2-day peak) and non-swimmers (no peak). With berm-open, we observed associations between GI and traditional and rapid methods for Enterococcus; fewer associations occurred when berm status was not considered. Conclusions: We found increased risk of GI at this urban runoff beach. When FIB source flowed freely (berm-open), several traditional and rapid indicators were related to illness. When FIB source was weak (berm-closed) fewer illness associations were seen. These different relationships under different conditions at a single beach demonstrate the difficulties using these indicators to predict health risk

Eliminating Summer Fallow Reduces Winter Wheat Yields, but Not Necessarily System Profitability

Summer fallow is commonly used to stabilize winter wheat (Triticum aestivum L.) production in the Central Great Plains, but summer fallow results in soil degradation, limits farm productivity and profitability, and stores soil water inefficiently. The objectives of this study were to quantify the production and economic consequences of replacing summer fallow with spring-planted crops on the subsequent winter wheat crop. A summer fallow treatment and five spring crop treatments [spring canola (Brassica napus L.), oat (Avena sativa L.) + pea (Pisum sativum L.) for forage, proso millet (Panicum miliaceum L.), dry bean (Phaseolus vulgaris L.), and corn (Zea mays L.)] were no-till seeded into sunflower (Helianthus annuus L.) residue in a randomized complete block design with five replications during 1999, 2000, and 2001. Winter wheat was planted in the fall following the spring crops. Five N fertilizer treatments (0, 22, 45, 67, and 90 kg N ha-1) were randomly assigned to each previous spring crop treatment in a split-plot treatment arrangement. The 3-yr mean wheat grain yield after summer fallow was 29% greater than following oat + pea for forage and 86% greater than following corn. The 3-yr mean annualized net return for the spring crop and subsequent winter wheat crop was $4.20, -$6.91, -$7.55, -$29.66, -$81.17, and -$94.88 ha-1 for oat + pea for forage, proso millet, summer fallow, dry bean, corn, and spring canola, respectively. Systems involving oat + pea for forage and proso millet are economically competitive with systems using summer fallow

EURODIAB childhood type 1 diabetes incidence registers - results from the first 20 years

Background and aims: The cellular properties of α cells in type 1 diabetes (T1D) are unknown. This is because T1D autoimmune destruction of β cells causes the islet mass to shrink in size rendering islet isolation and dispersion not technically feasible; and consequently electrophysiological characterization of islet cells to reveal the underling mechanisms explaining the distorted glucagon secretion in T1D could not be done. We employed GluCre-ROSA26-YFP (GYY) mice, which expresses YFP in pancreatic α cells. Along with our newly developed pancreas slice preparation whereby α cell and its precise secretory physiology within intact pancreatic tissue can be examined by patch clamp technique, unperturbed by conventional islet isolation and dispersion procedures, we are able to reliably localize and directly examine α cells electrophysiological properties in situ in health and T1D. We hypothesize that T1D α cells possess perturbed ion channels properties which contribute to hyperglucagonemia in early stage of T1D. Materials and methods: GYY mice were treated with streptozotocin (STZ) to induce T1D. IPGTT and radioimmunoassay (RIA) were performed to confirm diabetes phenotype. Pancreas slices were prepared from these mice to directly examine α cells ion channel properties in healthy and diseased islets by patch clamp technique. The identities of patched-cells were further confirmed by infusing fluoresecent marker (biocytin) during patching, showing its co-localization with YFP by confocal microscopy. Results: Normal GYY mice α cells in slices revealed identical electrophysiological features to those of their background C57/BL6 mice we previously characterized. These α cells are equipped with readily-activated A-type IK, voltage-gated INa, small size, low resting conductance, and inducible H/LVA ICa at -80mV. ICa influx correlated with glucagon exocytosis as either train of depolarization or UV photo-release of intracellular-loaded caged-Ca2+ stimulated Cm increase. 4 weeks after STZ treatment, GYY mice developed T1D, exhibiting higher fasting glucose, slower glucose clearance after a glucose challenge and higher fasting (control; 89 pg/ml vs. STZ group;122 pg/ml) and fed (control; 78 pg/ml vs. STZ group;112 pg/ml) serum glucagon levels. α cells in slices from these diabetic mice revealed augmentation of INa (control; 368 ± 43 pA vs. STZ group; 480 ± 71pA) and LVA ICa amplitudes (control; 40 ± 5 vs. STZ group; 49 ± 6 pA). HVA ICa however remained unaltered by T1D (control; 44 ± 5 pA vs. STZ group; 46 ± 6 pA) . Voltage-gated K+ current was found to be increased (STZ group; 2.13 nA vs. control; 1.76 nA). α cell size was unchanged compared to control (control; 4.7 ± 0.20 pF vs. STZ group; 4.8 ± 0.23 pF). Conclusion: GYY mouse α cell ion channel properties examined in slices were largely consistent with our previous findings and others, validating the feasibility of using pancreas slice approach to investigate α cells in normal and diabetic subjects. We postulate that the observed upregulation of INa and LVA ICa in diabetic α cells potentially elevates membrane potential that would more readily to trigger HVA Ca2+ channels opening, with ensuing initiation of action potential firing leading to glucagon secretion. This explains in part the observed glucagon hypersecretion in early stage of T1

The effects adiposity, diabetic status and depot-specificity on the activation of NFKB on JNK in human abdominal adipose tissue

Background and Aims: Both serum adiponectin, an adipokine with insulinsensitizing and anti-inflammatory action, and high sensitivity C-reactive protein (hsCRP), a marker of chronic low-grade systemic inflammation, have been reported to predict the development of type 2 diabetes. Here we examined, in a 10-year prospective study, whether these two biomarkers had an additive effect on predicting the long-term risk of type 2 diabetes. Materials and Methods: Serum adiponectin and hpCRP were measured in 417 non-diabetic Chinese subjects from the Hong Kong Cardiovascular Risk Factor Prevalence Study, who had returned for their 10-year follow-up in 2005/06. Of these, 76 had developed DM, according to the 1998 WHO diagnostic criteria. Adiponectin was measured using an in-house ELISA assay and hsCRP was measured using a particle-enhanced immunoturbidimetric assay. The association of adiponectin and hsCRP, alone or in combination, with the 10-year risk of diabetes was investigated. Results: Compared to the 341 subjects who remained non-diabetic at year- 10, subjects who had developed diabetes consisted of more men (p<0.003). They had higher baseline BMI, waist circumference, mean arterial blood pressure, plasma glucose (fasting and 2-hour post-OGTT), fasting insulin, HOMA-insulin resistance index (HOMA-IR), triglycerides, LDL-cholesterol and hsCRP levels (all p<0.001 except for LDL-cholesterol with p<0.05). They also had lower baseline serum adiponectin and HDL-cholesterol levels (both p<0.001). In a stepwise multiple logistic regression analysis model including sex, age, BMI, fasting insulin, presence of metabolic syndrome according to NCEP criteria, adiponectin and hsCRP, only baseline adiponectin (adjusted OR 0.44; 95% CI 0.27–0.74; p=0.002), hsCRP (adjusted OR 1.43; 95% CI 1.10–1.84; p=0.007) and metabolic syndrome (adjusted OR 2.72; 95% CI 1.55–4.78; p=0.001) were independently predictive of diabetes at 10 years. The addition of baseline adiponectin or hsCRP to a model consisting of sex, age and BMI significantly improved the prediction of diabetes risk, as reflected by the likelihood ratios in logistic regression analysis (p=0.001 for adiponectin and p=0.019 for hsCRP). Furthermore, the combined addition of adiponectin and hsCRP to the model provided significantly greater improvement than the addition of adiponectin alone (p=0.048) or the addition of hsCRP alone (p=0.003). Conclusion: Serum adiponectin and hsCRP were both independent predictors of the 10-year risk of diabetes in this Chinese cohort. Their usefulness as biomarkers for predicting the development of type 2 diabetes were over and above those of conventional risk factors including sex, age and BMI, and appeared to be additive when these two new biomarkers were used together. Supported by the STR seeding fund on Healthy Aging of the University of Hong Kong and the Innovation & Technology Fundlink_to_subscribed_fulltex

Long-term orlistat treatment reduces endotoxinaemia in fatty liver disease patients

Background and Aim: We previously tested the effects of a 18h culture in 2, 5, 10 and 30 mmol/l glucose (G2, G5, G10 and G30) on the transcriptome of cultured rat islets and identified 18 clusters with distinct glucose-dependent mRNA profiles. Of these, genes that were up-regulated between G10 and G30 are particularly interesting, as they may contribute to beta-cell glucotoxicity or protection against it. Besides genes involved in the unfolded protein response, this cluster contains most glycolytic enzymes and other genes typically induced by hypoxia, such as Adrenomedullin (Adm). Hypothesis: High glucose, which increases O2 consumption in β-cells, may induce hypoxia and thereby cause glucotoxicity. In this study, we tested whether overnight culture in high glucose activates the hypoxia-inducible transcription factor HIF in cultured rat beta-cells. Material and Methods: Male Wistar rat islets were precultured for one week in serum-free RPMI medium containing 10 mmol/l glucose (G10) and 5 g/l BSA, during which islets with signs of central necrosis were systematically discarded. INS1-E cells were cultured as monolayers (passage 72 to 80) in the presence of 10% FBS. Rat islets and INS1-E cells (70% confluence) were then cultured 18h in the presence of increasing glucose concentrations (G2, G5, G10 and G30) with various test substances and at different O2 concentrations (1%, 20% or 60% corresponding to a pO2 of 7.6, 152 and 456 mmHg). Results: Compared with G2, culture in G30 significantly increased 4-fold the protein levels of HIF1α and HIF2α, and 2-fold their dimerization partner HIF1β in INS1-E cell nuclear extracts while significantly decreasing HIF1β protein levels by ~50% in cytosolic extracts. High glucose also significantly up-regulated the mRNA levels of several HIF target genes (Adm, Glyceraldehyde phosphate dehydrogenase, Aldolase A, Lactate dehydrogenase A). These glucose effects, which were confirmed in whole rat islets, were mimicked by 6h exposure to a low pO2 (1% O2) or by 18h treatment with 10-30 µmol/l CoCl2, a known activator of HIF. In contrast, they were almost completely inhibited by 60% O2 or by various agents that inhibit Ca2+ influx and insulin secretion (250 µmol/l diazoxide, 1 µmol/l nimodipine and 1 µmol/l clonidine) and thereby likely reduce without suppressing the glucose stimulation of β-cell O2 consumption. In comparison with HIF target genes, Thioredoxin interacting protein (Txnip), one of the most glucose-responsive genes in cultured rat islets, was regulated in a completely different manner. Thus, Txnip mRNA levels were not increased by CoCl2 and 1% O2, and their increase by G30 was not reduced by 60% O2 and was markedly enhanced by diazoxide, nimodipine and clonidine. Conclusion: High glucose induces HIF1α and HIF2α protein stabilization, HIF1/2α-HIF1β dimer translocation to the nucleus, and increased expression of glycolytic enzymes and other HIF target genes in cultured rat beta-cells. These effects likely result from the glucose stimulation of ATP utilization and O2 consumption which may induce beta-cell hypoxia. These effects could contribute to in vitro β-cell glucotoxicity not only in whole islets but also in insulin-secreting cells cultured as monolayers

The correlation of dative bond length and parameter n in adducts Me3N-AlMe3-nCln (n

Complexes of the general formula Me3N-AlMe3-nCln (Me = CH3; n = 0, 1, 2, 3) 1-4 have been synthesized and structurally characterized by X-ray crystallography and NMR spectroscopy. The shape of the molecules is trigonal-antiprismatic with the nitrogen and aluminium atoms being tetrahedrally surrounded. The length of the dative bond N-Al is correlated with the parameter n due to inductive effects of the electronegative chlorine substituents, resulting in a difference in N-Al between 1 and 4 of 0.1 Angstrom (2.045, 2.010, 1.971, and 1.949 Angstrom). This shortening with increasing n, however, is not linear