43 Chipotle's remain closed as officials investigate E. coli outbreak

Twenty-two people in Washington and Oregon have been sickened; most of those people had eaten at Chipotle during past few weeks

From: http://www.cbsnews.com/news/43-chipotles-remain-closed-as-officials-investigate-e-coli-outbreak/

Year three of Obamacare sees spiked premiums for many

Health insurance sign-up season started Sunday for year three of the Affordable Care Act

From: http://www.cbsnews.com/videos/year-three-of-obamacare-sees-spiked-premiums-for-many/

In year 3, many hit with Obamacare sticker shock

Price of health insurance premiums goes down in a few states, but up dramatically in most, putting many buyers in tough positions

From: http://www.cbsnews.com/news/in-year-3-many-hit-with-obamacare-sticker-shock/

A novel exon in the human Ca2+-activated Cl- channel Ano1 imparts greater sensitivity to intracellular Ca2+

Anoctamin 1 (Ano1; TMEM16A) is a Ca²⁺-activated Cl^– channel (CACC) expressed in interstitial cells of Cajal. The mechanisms by which Ca²⁺ regulates Ano1 are incompletely understood. In the gastrointestinal tract, Ano1 is required for normal slow wave activity and is involved in regulating cell proliferation. Splice variants of Ano1 have varying electrophysiological properties and altered expression in disease states. Recently, we identified a transcript for human Ano1 containing a novel exon-"exon 0" upstream of and in frame with exon 1. The electrophysiological properties of this longer Ano1 isoform are unknown. Our aim was to determine the functional contribution of the newly identified exon to the Ca²⁺ sensitivity and electrophysiological properties of Ano1. Constructs with [Ano1(+0)] or without [Ano1(–0)] the newly identified exon were transfected into human embryonic kidney-293 cells. Voltage-clamp electrophysiology was used to determine voltage- and time-dependent parameters of whole cell Cl^– currents between isoforms with varying concentrations of intracellular Ca²⁺, extracellular anions, or Cl^– channel inhibitors. We found that exon 0 did not change voltage sensitivity and had no impact on the relative permeability of Ano1 to most anions. Ano1(+0) exhibited greater changes in current density but lesser changes in kinetics than Ano1(–0) in response to varying intracellular Ca²⁺. The CACC inhibitor niflumic acid inhibited current with greater efficacy and higher potency against Ano1(+0) compared with Ano1(–0). Likewise, the Ano1 inhibitor T16A_inh-A01 reduced Ano1(+0) more than Ano1(–0). In conclusion, human Ano1 containing exon 0 imparts its Cl^– current with greater sensitivity to intracellular Ca²⁺ and CACC inhibitors.

From: Strege, P. R., Bernard, C. E., Mazzone, A., Linden, D. R., Beyder, A., Gibbons, S. J., Farrugia, G. http://ajpgi.physiology.org/cgi/content/abstract/309/9/G743?rss=1

Zinc dyshomeostasis during polymicrobial sepsis in mice involves zinc transporter Zip14 and can be overcome by zinc supplementation

Integrity of the immune system is particularly dependent on the availability of zinc. Recent data suggest that zinc is involved in the development of sepsis, a life-threatening systemic inflammation with high death rates, but with limited therapeutic options. Altered cell zinc transport mechanisms could contribute to the inflammatory effects of sepsis. Zip14, a zinc importer induced by proinflammatory stimuli, could influence zinc metabolism during sepsis and serve as a target for therapy. Using cecal ligation-and-puncture (CLP) to model polymicrobial sepsis, we narrowed the function of ZIP14 to regulation of zinc homeostasis in hepatocytes, while hepatic leukocytes were mostly responsible for driving inflammation, as shown by higher expression of IL-1β, TNFα, S100A8, and matrix metalloproteinase-8. Using Zip14 knockout (KO) mice as a novel approach, we found that ablation of Zip14 produced a delay in development of leukocytosis, prevented zinc accumulation in the liver, altered the kinetics of hypozincemia, and drastically increased serum IL-6, TNFα, and IL-10 concentrations following CLP. Hence, this model revealed that the zinc transporter ZIP14 is a component of the pathway for zinc redistribution that contributes to zinc dyshomeostasis during polymicrobial sepsis. In contrast, using the identical CLP model, we found that supplemental dietary zinc reduced the severity of sepsis, as shown by amelioration of cytokines, calprotectins, and blood bacterial loads. We conclude that the zinc transporter ZIP14 influences aspects of the pathophysiology of nonlethal polymicrobial murine sepsis induced by CLP through zinc delivery. The results are promising for the use of zinc and its transporters as targets for future sepsis therapy.

From: Wessels, I., Cousins, R. J. http://ajpgi.physiology.org/cgi/content/abstract/309/9/G768?rss=1

Antibodies: friend or foe?

From: Bron, R., Bunnett, N. W. http://ajpgi.physiology.org/cgi/content/full/309/9/G717?rss=1

Dietary vitamin D3 deficiency alters intestinal mucosal defense and increases susceptibility to Citrobacter rodentium-induced colitis

Vitamin D deficiency affects more that 1 billion people worldwide. Although thought to increase risk of bacterial infections, the importance of vitamin D on host defense against intestinal bacterial pathogens is currently unclear since injection of the active form of vitamin D, 1,25(OH)2D3, increased susceptibility to the enteric bacterial pathogen Citrobacter rodentium by suppressing key immune/inflammatory factors. To further characterize the role of vitamin D during bacteria-induced colitis, we fed weanling mice either vitamin D3-deficient or vitamin D3-sufficient diets for 5 wk and then challenged them with C. rodentium. Vitamin D3-deficient mice lost significantly more body weight, carried higher C. rodentium burdens, and developed worsened histological damage. Vitamin D3-deficient mice also suffered greater bacterial translocation to extra-intestinal tissues, including mesenteric lymph nodes, spleen, and liver. Intestinal tissues of infected vitamin D3-deficient mice displayed increased inflammatory cell infiltrates as well as significantly higher gene transcript levels of inflammatory mediators TNF-α, IL-1β, IL-6, TGF-β, IL-17A, and IL-17F as well as the antimicrobial peptide REG3. Notably, these exaggerated inflammatory responses accelerated the loss of commensal microbes and were associated with an impaired ability to detoxify bacterial lipopolysaccharide. Overall, these studies show that dietary-induced vitamin D deficiency exacerbates intestinal inflammatory responses to infection, also impairing host defense.

From: Ryz, N. R., Lochner, A., Bhullar, K., Ma, C., Huang, T., Bhinder, G., Bosman, E., Wu, X., Innis, S. M., Jacobson, K., Vallance, B. A. http://ajpgi.physiology.org/cgi/content/abstract/309/9/G730?rss=1

Chronic alcohol exposure affects pancreatic acinar mitochondrial thiamin pyrophosphate uptake: studies with mouse 266-6 cell line and primary cells

Thiamin is essential for normal metabolic activity of all mammalian cells, including those of the pancreas. Cells obtain thiamin from their surroundings and enzymatically convert it into thiamin pyrophosphate (TPP) in the cytoplasm; TPP is then taken up by mitochondria via a specific carrier the mitochondrial TPP transporter (MTPPT; product of the SLC25A19 gene). Chronic alcohol exposure negatively impacts the health of pancreatic acinar cells (PAC), but its effect on physiological/molecular parameters of MTPPT is not known. We addressed this issue using mouse pancreatic acinar tumor cell line 266-6 and primary PAC of wild-type and transgenic mice carrying the SLC25A19 promoter that were fed alcohol chronically. Chronic alcohol exposure of 266-6 cells (but not to its nonoxidative metabolites ethyl palmitate and ethyl oleate) led to a significant inhibition in mitochondrial TPP uptake, which was associated with a decreased expression of MTPPT protein, mRNA, and activity of the SLC25A19 promoter. Similarly, chronic alcohol feeding of mice led to a significant inhibition in expression of MTPPT protein, mRNA, heterogeneous nuclear RNA, as well as in activity of SLC25A19 promoter in PAC. While chronic alcohol exposure did not affect DNA methylation of the Slc25a19 promoter, a significant decrease in histone H3 euchromatin markers and an increase in H3 heterochromatin marker were observed. These findings show, for the first time, that chronic alcohol exposure negatively impacts pancreatic MTPPT, and that this effect is exerted, at least in part, at the level of Slc25a19 transcription and appears to involve epigenetic mechanism(s).

From: Srinivasan, P., Nabokina, S., Said, H. M. http://ajpgi.physiology.org/cgi/content/abstract/309/9/G750?rss=1

Plasma miR-185 is decreased in patients with esophageal squamous cell carcinoma and might suppress tumor migration and invasion by targeting RAGE

The receptor for advanced-glycation end products (RAGE) is upregulated in various cancers and has been associated with tumor progression, but little is known about its expression and regulation by microRNAs (miRNAs) in esophageal squamous cell carcinoma (ESCC). Here, we describe miR-185, which represses RAGE expression, and investigate the biological role of miR-185 in ESCC. In this study, we found that the high level of RAGE expression in 29 pairs of paraffin-embedded ESCC tissues was correlated positively with the depth of invasion by immunohistochemistry, suggesting that RAGE was involved in ESCC. We used bioinformatics searches and luciferase reporter assays to investigate the prediction that RAGE was regulated directly by miR-185. Besides, overexpression of miR-185 in ESCC cells was accompanied by 27% (TE-11) and 49% (Eca-109) reduced RAGE expression. The effect was further confirmed in RAGE protein by immunofluorescence in both cell lines. The effects were reversed following cotransfection with miR-185 and high-level expression of the RAGE vector. Furthermore, the biological role of miR-185 in ESCC cell lines was investigated using assays of cell viability, Ki-67 staining, and cell migration and invasion, as well as in a xenograft model. We found that overexpression of miR-185 inhibited migration and invasion by ESCC cells in vitro and reduced their capacity to develop distal pulmonary metastases in vivo partly through the RAGE/heat shock protein 27 pathway. Interestingly, in clinical specimens, the level of plasma miR-185 expression was decreased significantly (P = 0.002) in patients with ESCC [0.500; 95% confidence interval (CI) 0.248–1.676] compared with healthy controls (2.410; 95% CI 0.612–5.671). The value of the area under the receiver-operating characteristic curve was 0.73 (95% CI 0.604–0.855). In conclusion, our findings shed novel light on the role of miR-185/RAGE in ESCC metastasis, and plasma miR-185 has potential as a novel diagnostic biomarker in ESCC.

From: Jing, R., Chen, W., Wang, H., Ju, S., Cong, H., Sun, B., Jin, Q., Chu, S., Xu, L., Cui, M. http://ajpgi.physiology.org/cgi/content/abstract/309/9/G719?rss=1

Effect of dietary fructose on portal and systemic serum fructose levels in rats and in KHK-/- and GLUT5-/- mice

Elevated blood fructose concentrations constitute the basis for organ dysfunction in fructose-induced metabolic syndrome. We hypothesized that diet-induced changes in blood fructose concentrations are regulated by ketohexokinase (KHK) and the fructose transporter GLUT5. Portal and systemic fructose concentrations determined by HPLC in wild-type mice fed for 7 days 0% free fructose were <0.07 mM, were independent of time after feeding, were similar to those of GLUT5^–/–, and did not lead to hyperglycemia. Postprandial fructose levels, however, increased markedly in those fed isocaloric 20% fructose, causing significant hyperglycemia. Deletion of KHK prevented fructose-induced hyperglycemia, but caused dramatic hyperfructosemia (>1 mM) with reversed portal to systemic gradients. Systemic fructose in wild-type and KHK^–/– mice changed by 0.34 and 1.8 mM, respectively, for every millimolar increase in portal fructose concentration. Systemic glucose varied strongly with systemic, but not portal, fructose levels in wild-type, and was independent of systemic and portal fructose in KHK^–/–, mice. With ad libitum feeding for 12 wk, fructose-induced hyperglycemia in wild-type, but not hyperfructosemia in KHK^–/– mice, increased HbA_1c concentrations. Increasing dietary fructose to 40% intensified the hyperfructosemia of KHK^–/– and the fructose-induced hyperglycemia of wild-type mice. Fructose perfusion or feeding in rats also caused duration- and dose-dependent hyperfructosemia and hyperglycemia. Significant levels of blood fructose are maintained independent of dietary fructose, KHK, and GLUT5, probably by endogenous synthesis of fructose. KHK prevents hyperfructosemia and fructose-induced hyperglycemia that would markedly increase HbA_1c levels. These findings explain the hyperfructosemia of human hereditary fructosuria as well as the hyperglycemia of fructose-induced metabolic syndrome.

From: Patel, C., Sugimoto, K., Douard, V., Shah, A., Inui, H., Yamanouchi, T., Ferraris, R. P. http://ajpgi.physiology.org/cgi/content/abstract/309/9/G779?rss=1

RhoA enhances store-operated Ca2+ entry and intestinal epithelial restitution by interacting with TRPC1 after wounding

Early mucosal restitution occurs as a consequence of epithelial cell migration to resealing of superficial wounds after injury. Our previous studies show that canonical transient receptor potential-1 (TRPC1) functions as a store-operated Ca²⁺ channel (SOC) in intestinal epithelial cells (IECs) and plays an important role in early epithelial restitution by increasing Ca²⁺ influx. Here we further reported that RhoA, a small GTP-binding protein, interacts with and regulates TRPC1, thus enhancing SOC-mediated Ca²⁺ entry (SOCE) and epithelial restitution after wounding. RhoA physically associated with TRPC1 and formed the RhoA/TRPC1 complexes, and this interaction increased in stable TRPC1-transfected IEC-6 cells (IEC-TRPC1). Inactivation of RhoA by treating IEC-TRPC1 cells with exoenzyme C₃ transferase (C₃) or ectopic expression of dominant negative RhoA (DNMRhoA) reduced RhoA/TRPC1 complexes and inhibited Ca²⁺ influx after store depletion, which was paralleled by an inhibition of cell migration over the wounded area. In contrast, ectopic expression of wild-type (WT)-RhoA increased the levels of RhoA/TRPC1 complexes, induced Ca²⁺ influx through activation of SOCE, and promoted cell migration after wounding. TRPC1 silencing by transfecting stable WT RhoA-transfected cells with siRNA targeting TRPC1 (siTRPC1) reduced SOCE and repressed epithelial restitution. Moreover, ectopic overexpression of WT-RhoA in polyamine-deficient cells rescued the inhibition of Ca²⁺ influx and cell migration induced by polyamine depletion. These findings indicate that RhoA interacts with and activates TRPC1 and thus stimulates rapid epithelial restitution after injury by inducing Ca²⁺ signaling.

From: Chung, H. K., Rathor, N., Wang, S. R., Wang, J.-Y., Rao, J. N. http://ajpgi.physiology.org/cgi/content/abstract/309/9/G759?rss=1

Eat Well, America!

This American Diabetes Month, We’re Helping you Eating Well, America^SM!

November is American Diabetes Month. That means it’s time to Eat Well, America! Whether you are one of the nearly 30 million Americans living with diabetes or the 86 million Americans with prediabetes or you simply want to live a healthier lifestyle—our Eat Well, America! campaign will show you how easy and joyful healthy eating can be.

Throughout November, we will share nutritious recipes selected by noted chefs and cookbook authors for every meal, including snacks and special occasion treats.

What’s In it For Me?!

• Delicious breakfast recipes that motivate you to get your busy day off to the right start.
• Easy and satisfying snack recipes that make the walk past the vending machine a breeze.
• Lunch recipes to celebrate National Healthy Lunch Day on Nov. 17.
• Seasonal dinner recipes that ensure you don’t miss out on the autumn and holiday flavors you love.
• Healthy special occasion options, including sweets, so you never feel deprived of your favorite treats and stay on track with good nutrition.

How Can I Participate?

1. Follow us on Facebook, Twitter, Instagram, Pinterest and YouTube throughout November!
2. Join the #EatWellAmerica conversation on social media! Share your own favorite healthy recipes and meal preparation tips with your friends and family.
3. Celebrate #HealthyLunchDay on Nov. 17. Bring or buy a healthy lunch and share your plate on social media using #MyHealthyLunch. Remember to tag us in your post!
4. Join our #HealthyLunchDay Thunderclap and donate your social media status to ignite a dialogue about the importance of healthy eating!

We look forward to putting good food, and good health, on your table this month!

From: American Diabetes Association http://diabetesstopshere.org/2015/11/01/eat-well-america/