Visfatin reduces gap junction mediated cell-to-cell communication in proximal tubule-derived epithelial cells

Background/Aims: In the current study we examined if the adipocytokine, visfatin, alters connexin-mediated intercellular communication in proximal tubule-derived epithelial cells. Methods: The effects of visfatin (10-200ng/mL) on cell viability and cytotoxicity in HK2-cells were assessed by MTT, crystal violet and lactate dehydrogenase assays. Western blot analysis was used to confirm expression of Cx26, Cx40 and Cx43. The effect of visfatin (10-200ng/mL) on TGF-β1 secretion was confirmed by ELISA, and the effects of both TGF-β1 (2-10ng/mL) and visfatin (10-200ng/mL) on connexin expression were assessed by western blot. Functional intercellular communication was determined using transfer of Lucifer Yellow and paired-whole cell patch clamp electrophysiology. Results: In low glucose (5mM), visfatin (10-200ng/mL) did not affect membrane integrity, cytotoxicity or cell viability at 48hrs, but did evoke a concentration-dependent reduction in Cx26 and Cx43 expression. The expression of Cx40 was unaffected. At 48hrs, visfatin (10-200ng/mL) increased the secretion of TGF-β1 and the visfatin-evoked changes in connexin expression were mimicked by exogenous application of the pro-fibrotic cytokine (2-10ng/ml). Visfatin reduced dye transfer between coupled cells and decreased functional conductance, with levels falling by 63% as compared to control. Although input resistance was increased following visfatin treatment by 166%, the change was not significant as compared to control. The effects of visfatin on Cx-expression and cell-coupling were blocked in the presence of a TGF-β1 specific neutralizing antibody. Conclusions: The adipocytokine visfatin selectively evoked a non-toxic reduction in connexin expression in HK2-cells. The loss in gap-junction associated proteins was mirrored by a loss in functional conductance between coupled cells. Visfatin increased TGF-β secretion and the pattern of change for connexins expression was mimicked by exogenous application of TGF-β1. The effect of visfatin on Cx-expression and dye transfer were negated in the presence of a TGF-β1 neutralising antibody. These data suggest that visfatin reduces connexin-mediated intercellular communication in proximal tubule-derived epithelial cells via a TGF-β dependent pathway. © 2013 S. Karger AG, Base

Adenosine A1 receptor activation mediates the developmental shift at layer 5 pyramidal cell synapses and is a determinant of mature synaptic strength

During the first postnatal month glutamatergic synapses between layer 5 pyramidal cells in the rodent neocortex switch from an immature state exhibiting high probability of neurotransmitter release, large unitary amplitude and synaptic depression to a mature state with decreased probability of release, smaller unitary amplitude and synaptic facilitation. Using paired recordings, we demonstrate that the developmental shift in release probability at synapses between rat somatosensory layer 5 thick-tufted pyramidal cells is due to a higher and more heterogeneous activation of presynaptic adenosine A1 receptors. Immature synapses under control conditions exhibited distributions of CV, failure rate and release probability that were almost coincident with the A1 receptor blocked condition; however, mature synapses under control conditions exhibited much broader distributions that spanned those of both the A1 receptor agonised and antagonised conditions. Immature and mature synapses expressed A1 receptors with no observable difference in functional efficacy and therefore the heterogeneous A1 receptor activation seen in the mature neocortex is due to increased adenosine concentrations that vary between synapses. Given the central role demonstrated for A1 receptor activation in determining synaptic amplitude and the statistics of transmission between mature layer 5 pyramidal cells, the emplacement of adenosine sources and sinks near the synaptic terminal could constitute a novel form of long-term synaptic plasticity

Growth and Performance of Nursery Pigs Fed Crude Glycerol

The growth and performance of 96 nursery pigs fed crude glycerol was evaluated in a 33 d trial. Pigs were weaned at 21 d of age and had an average start weight of 7.9±1.2 kg (17.4±2.6 lb). Pigs were provided ad libitum access to corn soybean diets containing three levels of crude glycerol, 0, 5, or 10%. Diets were formulated to be ioscaloric and isolysinic. There was no difference in pig performance based on dietary treatment. Crude glycerol can be fed to young pigs as an energy source without growth or performance effects

Growth Performance of Growing Pigs Fed Crude Glycerol-Supplemented Diets

Growth performance of growing pigs fed crude glycerol was determined in a 138-d feeding trial. Crude glycerol utilized in the trial contained 84.51% glycerol, 11.95% water, 2.91% sodium chloride, and 0.32% methanol. Eight days post-weaning, 96 pigs (48 barrows, 48 gilts) with an average BW of 7.9 ± 0.4 kg were allotted to 24 pens (4 pigs/pen), with gender and pen weight balanced at the start of the experiment. Dietary regimes were randomly assigned to each pen. Dietary treatments were 0, 5, and 10% crude glycerol inclusion in corn-soybean meal based diets. Diets were offered ad libitum in meal form and formulated to be equal in metabolizable energy (ME), sodium, chloride, and Lys, with other amino acids (AA) balanced on an ideal AA basis. Every two weeks, pigs and feeders were weighed and G:F calculated. Pig growth, feed intake, and G:F were not affected by dietary treatment. Crude glycerol is a viable source of dietary energy that is well utilized by pigs. Inclusion of crude glycerol in pig diets may be determined by relative availability and price of other dietary energy sources

Electronic health record phenotyping improves detection and screening of type 2 diabetes in the general United States population: A cross-sectional, unselected, retrospective study

Objectives: In the United States, 25% of people with type 2 diabetes are undiagnosed. Conventional screening models use limited demographic information to assess risk. We evaluated whether electronic health record (EHR) phenotyping could improve diabetes screening, even when records are incomplete and data are not recorded systematically across patients and practice locations. Methods: In this cross-sectional, retrospective study, data from 9,948 US patients between 2009 and 2012 were used to develop a pre-screening tool to predict current type 2 diabetes, using multivariate logistic regression. We compared (1) a full EHR model containing prescribed medications, diagnoses, and traditional predictive information, (2) a restricted EHR model where medication information was removed, and (3) a conventional model containing only traditional predictive information (BMI, age, gender, hypertensive and smoking status). We additionally used a random-forests classification model to judge whether including additional EHR information could increase the ability to detect patients with Type 2 diabetes on new patient samples. Results: Using a patient's full or restricted EHR to detect diabetes was superior to using basic covariates alone (p<0.001). The random forests model replicated on out-of-bag data. Migraines and cardiac dysrhythmias were negatively associated with type 2 diabetes, while acute bronchitis and herpes zoster were positively associated, among other factors. Conclusions: EHR phenotyping resulted in markedly superior detection of type 2 diabetes in a general US population, could increase the efficiency and accuracy of disease screening, and are capable of picking up signals in real-world records

Cross-sectional associations between domain-specific sitting time and other lifestyle health behaviours: the Stormont study

BackgroundThere is a dearth of literature on how different domains of sitting time relate to other health behaviours. Therefore, this study aimed to explore these associations in a sample of office workers.Methods7170 Northern Irish Civil Servants completed an online survey which included information on workday and non-workday sitting time in five domains (travel, work, TV, computer-use, leisure-time), physical activity, fruit and vegetable intake, alcohol consumption and cigarette smoking. An unhealthy behaviour score was calculated by summing the number of health behaviours which did not meet the current guidelines. Multinomial regressions examined associations between unhealthy behaviour score and each domain of sitting time.Results≥7 hours sitting at work and ≥2 hours TV viewing on a workday both more than doubled the odds of partaking in ≥3 unhealthy behaviours [Odds ratio, OR = 2.03, 95% CI, (1.59–2.61); OR = 2.19 (1.71–2.80)] and ≥3 hours of TV viewing on a non-workday nearly tripled the odds [OR = 2.96 (2.32–3.77)].ConclusionsHigh sitting time at work and TV viewing on a workday and non-workday are associated with increased odds of partaking in multiple unhealthy behaviours. Interventions need to focus on these domains and public health policy should consider sitting time as an important health behaviour

Estimated Drug Overdose Deaths Averted by North America's First Medically-Supervised Safer Injection Facility

Illicit drug overdose remains a leading cause of premature mortality in urban settings worldwide. We sought to estimate the number of deaths potentially averted by the implementation of a medically supervised safer injection facility (SIF) in Vancouver, Canada.The number of potentially averted deaths was calculated using an estimate of the local ratio of non-fatal to fatal overdoses. Inputs were derived from counts of overdose deaths by the British Columbia Vital Statistics Agency and non-fatal overdose rates from published estimates. Potentially-fatal overdoses were defined as events within the SIF that required the provision of naloxone, a 911 call or an ambulance. Point estimates and 95% Confidence Intervals (95% CI) were calculated using a Monte Carlo simulation. Between March 1, 2004 and July 1, 2008 there were 1004 overdose events in the SIF of which 453 events matched our definition of potentially fatal. In 2004, 2005 and 2006 there were 32, 37 and 38 drug-induced deaths in the SIF's neighbourhood. Owing to the wide range of non-fatal overdose rates reported in the literature (between 5% and 30% per year) we performed sensitivity analyses using non-fatal overdose rates of 50, 200 and 300 per 1,000 person years. Using these model inputs, the number of averted deaths were, respectively: 50.9 (95% CI: 23.6–78.1); 12.6 (95% CI: 9.6–15.7); 8.4 (95% CI: 6.5–10.4) during the study period, equal to 1.9 to 11.7 averted deaths per annum.Based on a conservative estimate of the local ratio of non-fatal to fatal overdoses, the potentially fatal overdoses in the SIF during the study period could have resulted in between 8 and 51 deaths had they occurred outside the facility, or from 6% to 37% of the total overdose mortality burden in the neighborhood during the study period. These data should inform the ongoing debates over the future of the pilot project

The First Substellar Subdwarf? Discovery of a Metal-poor L Dwarf with Halo Kinematics

We present the discovery of the first L-type subdwarf, 2MASS J05325346+8246465. This object exhibits enhanced collision-induced H$_2$ absorption, resulting in blue NIR colors ($J-K_s = 0.26{\pm}0.16$). In addition, strong hydride bands in the red optical and NIR, weak TiO absorption, and an optical/J-band spectral morphology similar to the L7 DENIS 0205$-$1159AB imply a cool, metal-deficient atmosphere. We find that 2MASS 0532+8246 has both a high proper motion, $\mu$ = 2\farcs60\pm0\farcs15 yr$^{-1}$, and a substantial radial velocity, $v_{rad} = -195{\pm}11$ km s$^{-1}$, and its probable proximity to the Sun (d = 10--30 pc) is consistent with halo membership. Comparison to subsolar-metallicity evolutionary models strongly suggests that 2MASS 0532+8246 is substellar, with a mass of 0.077 $\lesssim$ M $\lesssim$ 0.085 M_{\sun} for ages 10--15 Gyr and metallicities $Z = 0.1-0.01$ Z_{\sun}. The discovery of this object clearly indicates that star formation occurred below the Hydrogen burning mass limit at early times, consistent with prior results indicating a flat or slightly rising mass function for the lowest-mass stellar subdwarfs. Furthermore, 2MASS 0532+8246 serves as a prototype for a new spectral class of subdwarfs, additional examples of which could be found in NIR proper motion surveys.Comment: 9 pages, 3 figures, accepted to Ap