Absence of neurotensin attenuates intestinal dysbiosis and inflammation by maintaining Mmp7/α-defensin axis in diet-induced obese mice.

We previously reported that high levels of plasma neurotensin (NT), a gut hormone released from enteroendocrine cells of the small bowel, contribute to obesity and comorbid conditions. Gut microbiota has been implicated in the obesity development. Paneth cells are critical in maintaining gut microbiota composition and homeostasis by releasing antimicrobial proteins including α-defensins. The purpose of our current study was to determine the possible role of NT in gut microbiota composition and α-defensin gene expression associated with obesity. Here we show that the ratio of Firmicutes/Bacteroidetes (F/B ratio) and intestinal proinflammatory cytokines is significantly increased in NT+/+ mice fed with a high-fat diet (HFD) which were improved in NT-deficient mice. HFD disrupted the intestinal Mmp7/α-defensin axis, which was completely prevented in NT-/- mice. In addition, NT treatment inhibited DEFA5 expression and concurrent NF-κB activity, which was blocked by a pan PKC inhibitor (Gö6983) or an inhibitor for atypical PKCs (CRT0066854). More importantly, the shRNA-mediated knockdown of atypical PKCτ reversed NT-attenuated DEFA5 expression and increased NF-κB activity. NT contributes to the HFD-induced disruption of gut microbiota composition and α-defensin expression. PKCτ/λ plays a central role in NT-mediated α-defensin gene expression which might be mediated through the inhibition of NF-κB signaling pathways in Paneth cells.

FFAR4 Is Involved in Regulation of Neurotensin Release From Neuroendocrine Cells and Male C57BL/6 Mice.

Neurotensin (NT), a 13 amino-acid peptide, is predominantly released from enteroendocrine cells of the small bowel in response to fat ingestion. Free fatty acid receptors (FFARs) FFAR1 and FFAR4 regulate secretion of gut hormones and insulin. Here, we show that docosahexaenoic acid, a long-chain fatty acid, has the most dramatic effect on NT release. FFAR1 agonists slightly stimulate and FFAR4 agonists dramatically stimulate and amplify NT secretion. Double knockdown of FFAR1 and FFAR4 decreases NT release, whereas overexpression of FFAR4, but not FFAR1, increases NT release. Administration of cpdA, an FFAR4 agonist, but not TAK-875, a selective FFAR1 agonist, increases plasma NT levels and further increases olive oil-stimulated plasma NT levels. Inhibition of MAPK kinase (MEK)/ERK1/2 decreased fatty acid-stimulated NT release but increased AMP-activated protein kinase (AMPK) phosphorylation. In contrast, inhibition of AMPK further increased NT secretion and ERK1/2 phosphorylation mediated by FFAR1 or FFAR4. Our results indicate that FFAR4 plays a more critical role than FFAR1 in mediation of fat-regulated NT release and in inhibitory crosstalk between MEK/ERK1/2 and AMPK in the control of NT release downstream of FFAR1 and FFAR4.

Arytenoid abduction for dynamic rehabilitation of bilateral laryngeal paralysis.

OBJECTIVES: Bilateral laryngeal paralysis results in airway obstruction, but the voice is often nearly normal. Tracheotomy provides an airway and preserves voice. Surgical procedures to statically enlarge the glottis can permit decannulation, but do so at the expense of the voice. Motion analysis in cadaver larynges has demonstrated that adductor and abductor muscles rotate the arytenoid cartilage around different axes. We sought to determine whether external rotation of the arytenoid cartilage could enlarge the airway without abolishing residual phonatory adduction. METHODS: We performed arytenoid abduction in 6 patients with obstructing laryngeal paralysis. A suture was placed in the muscular process and posterior-inferior traction was applied, anchoring the suture to the inferior cornu of the thyroid cartilage. Outcomes were evaluated by assessing airway symptoms, by assessing the voice, and by documentation of laryngeal motion via videolaryngoscopy. RESULTS: Three patients with severe stridor had marked relief of symptoms, and 2 of the 3 tracheotomy-dependent patients were decannulated. Three patients had good voices, 2 had mild breathiness, and 1 was very breathy. CONCLUSIONS: Arytenoid abduction is a promising treatment for relieving airway obstruction in patients with laryngeal paralysis. It has the potential to preserve voice in patients with residual phonatory adduction.

Activation of AMPK Stimulates Neurotensin Secretion in Neuroendocrine Cells.

AMP-activated protein kinase (AMPK), a critical fuel-sensing enzyme, regulates the metabolic effects of various hormones. Neurotensin (NT) is a 13-amino acid peptide predominantly localized in enteroendocrine cells of the small bowel and released by fat ingestion. Increased fasting plasma levels of pro-NT (a stable NT precursor fragment produced in equimolar amounts relative to NT) are associated with an increased risk of diabetes, cardiovascular disease, and mortality; however, the mechanisms regulating NT release are not fully defined. We previously reported that inhibition of the mammalian target of rapamycin (mTOR) complex 1 (mTORC1) increases NT secretion and gene expression through activation of the MEK/ERK pathway. Here, we show that activation of AMPK increases NT secretion from endocrine cell lines (BON and QGP-1) and isolated mouse crypt cells enriched for NT-positive cells. In addition, plasma levels of NT increase in mice treated with 5-aminoimidazole-4-carboxamide riboside, a pharmacologic AMPK activator. Small interfering RNA-mediated knockdown of AMPKα decrease, whereas overexpression of the subunit significantly enhances, NT secretion from BON cells treated with AMPK activators or oleic acid. Similarly, small interfering RNA knockdown of the upstream AMPK kinases, liver kinase B1 and Ca(2+) calmodulin-dependent protein kinase kinase 2, also attenuate NT release and AMPK phosphorylation. Moreover, AMPK activation increases NT secretion through inhibition of mTORC1 signaling. Together, our findings show that AMPK activation enhances NT release through inhibition of mTORC1 signaling, thus demonstrating an important cross talk regulation for NT secretion.

The association of tongue scalloping with obstructive sleep apnea and related sleep pathology.

OBJECTIVE: The association between OSAS and patient history and physical exam findings is previously established; however, to our knowledge there are no studies that evaluate the role of tongue scalloping as a reliable clinical indicator for OSA, snoring, or the presence of other sleep pathology as evidenced by polysomnography. This study evaluates the hypothesis that such an association exists. SUBJECTS AND METHODS: Sixty-one otolaryngology clinic patients were evaluated by history and physical exam for the presence and degree of tongue scalloping, snoring, and other previously established clinical indicators for sleep-disordered breathing and obstructive apnea. Twenty-five of the 61 study patients were additionally evaluated by overnight polysomnography to provide conclusive diagnosis of sleep pathology. The degree of tongue scalloping was graded from 0 to 3 and its significance as a screening, diagnostic, and predictive factor for sleep pathology was then statistically determined. RESULTS: Twenty-seven patients (44%) had known or newly documented OSA and 47 (77%) had a history of snoring. Twenty-seven patients (44%) had some degree of tongue scalloping (1-3) and 74% of these patients were male. The presence of any degree of tongue scalloping (grade 1-3) in patients with known or newly documented OSA showed sensitivity, specificity, PPV, and NPV of 52%, 68%, 70%, and 50% respectively. The presence of tongue scalloping in patients with either known snoring history or newly documented snoring showed sensitivity, specificity, PPV, and NPV of 47%, 64%, 81%, and 26% respectively. Presence of tongue scalloping was 71% specific for abnormal sleep efficiency (<85%), 70% specific for abnormal AHI (>5), and 86% specific for nocturnal desaturation >4% below baseline. Presence of tongue scalloping also showed PPV of 67% for abnormal AHI, 89% for apnea or hypopnea, and 89% for nocturnal desaturation. Presence and severity of tongue scalloping showed positive correlation with increasing Mallampati and modified Mallampati airway classification. CONCLUSIONS: In high-risk patients we found tongue scalloping to be predictive of sleep pathology. Tongue scalloping was also associated with pathologic polysomnography data and abnormal Mallampati grades. We feel the finding of tongue scalloping is a useful clinical indicator of sleep pathology and that its presence should prompt the physician to inquire about snoring history.