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1.
Int J Biol Macromol ; 208: 299-313, 2022 May 31.
Artículo en Inglés | MEDLINE | ID: mdl-35288166

RESUMEN

The innate cartilage extracellular matrix is avascular and plays a vital role in innate chondrocytes. Recapping the crucial components of the extracellular matrix in engineered organs via polymeric gels and bioinspired approaches is promising for improving the regenerative aptitude of encapsulated cartilage/chondrocytes. Conventional gel formation techniques for polymeric materials rely on employing oxidative crosslinking, which is constrained in this avascular environment. Further, poor mechanical properties limit the practical applications of polymeric gels and reduce their therapeutic efficacy. Herein, the purpose of this study was to develop a bioadhesive gel possessing dual crosslinking for engineering cartilage. Tyramine (TYR) was first chemically conjugated to the alginate (ALG) backbone to form an ALG-TYR precursor, followed by the addition of calcium peroxide (CaO2); calcium ions of CaO2 physically crosslink with ALG, and oxygen atoms of CaO2 chemically crosslink TYR with tyrosinase, thus enabling dual/enhanced crosslinking and possessing injectability. The ALG-TYR/tyrosinase/CaO2 gel system was chemically, mechanically, cellularly, and microscopically characterized. The gel system developed herein was biocompatible and showed augmented mechanical strength. The results showed, for the first time, that CaO2 supplementation preserved cell viability and enhanced the crosslinking ability, bioadhesion, mechanical strength, chondrogenesis, and stability for cartilage regeneration.


Asunto(s)
Alginatos , Monofenol Monooxigenasa , Alginatos/química , Cartílago , Condrocitos , Condrogénesis , Hidrogeles/química , Peróxidos , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Tiramina
2.
iScience ; 24(7): 102807, 2021 Jul 23.
Artículo en Inglés | MEDLINE | ID: mdl-34337362

RESUMEN

Small-molecule tankyrase 1 and tankyrase 2 (TNKS1/2) inhibitors are effective antitumor agents in selected tumor cell lines and mouse models. Here, we characterized the response signatures and the in-depth mechanisms for the antiproliferative effect of tankyrase inhibition (TNKSi). The TNKS1/2-specific inhibitor G007-LK was used to screen 537 human tumor cell lines and a panel of particularly TNKSi-sensitive tumor cell lines was identified. Transcriptome, proteome, and bioinformatic analyses revealed the overall TNKSi-induced response signatures in the selected panel. TNKSi-mediated inhibition of wingless-type mammary tumor virus integration site/ß-catenin, yes-associated protein 1 (YAP), and phosphatidylinositol-4,5-bisphosphate 3-kinase/AKT signaling was validated and correlated with lost expression of the key oncogene MYC and impaired cell growth. Moreover, we show that TNKSi induces accumulation of TNKS1/2-containing ß-catenin degradasomes functioning as core complexes interacting with YAP and angiomotin proteins during attenuation of YAP signaling. These findings provide a contextual and mechanistic framework for using TNKSi in anticancer treatment that warrants further comprehensive preclinical and clinical evaluations.

3.
Cell Metab ; 31(6): 1173-1188.e5, 2020 06 02.
Artículo en Inglés | MEDLINE | ID: mdl-32413335

RESUMEN

G protein-coupled receptor 120 (GPR120) and PPARγ agonists each have insulin sensitizing effects. But whether these two pathways functionally interact and can be leveraged together to markedly improve insulin resistance has not been explored. Here, we show that treatment with the PPARγ agonist rosiglitazone (Rosi) plus the GPR120 agonist Compound A leads to additive effects to improve glucose tolerance and insulin sensitivity, but at lower doses of Rosi, thus avoiding its known side effects. Mechanistically, we show that GPR120 is a PPARγ target gene in adipocytes, while GPR120 augments PPARγ activity by inducing the endogenous ligand 15d-PGJ2 and by blocking ERK-mediated inhibition of PPARγ. Further, we used macrophage- (MKO) or adipocyte-specific GPR120 KO (AKO) mice to show that GRP120 has anti-inflammatory effects via macrophages while working with PPARγ in adipocytes to increase insulin sensitivity. These results raise the prospect of a safer way to increase insulin sensitization in the clinic.


Asunto(s)
Insulina/metabolismo , PPAR gamma/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Células 3T3-L1 , Acetatos/farmacología , Adipocitos/metabolismo , Animales , Células Cultivadas , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , PPAR gamma/agonistas , Receptores Acoplados a Proteínas G/agonistas , Receptores Acoplados a Proteínas G/deficiencia , Rosiglitazona/farmacología , Tiramina/análogos & derivados , Tiramina/farmacología
4.
Int J Obes (Lond) ; 44(8): 1691-1702, 2020 08.
Artículo en Inglés | MEDLINE | ID: mdl-32317752

RESUMEN

OBJECTIVE: Human TNKS, encoding tankyrase 1 (TNKS1), localizes to a susceptibility locus for obesity and type 2 diabetes mellitus (T2DM). Here, we addressed the therapeutic potential of G007-LK, a TNKS-specific inhibitor, for obesity and T2DM. METHODS: We administered G007-LK to diabetic db/db mice and measured the impact on body weight, abdominal adiposity, and serum metabolites. Muscle, liver, and white adipose tissues were analyzed by quantitative RT-PCR and western blotting to determine TNKS inhibition, lipolysis, beiging, adiponectin level, mitochondrial oxidative metabolism and mass, and gluconeogenesis. Protein interaction and PARylation analyses were carried out by immunoprecipitation, pull-down and in situ proximity ligation assays. RESULTS: TNKS inhibition reduced body weight gain, abdominal fat content, serum cholesterol levels, steatosis, and proteins associated with lipolysis in diabetic db/db mice. We discovered that TNKS associates with PGC-1α and that TNKS inhibition attenuates PARylation of PGC-1α, contributing to increased PGC-1α level in WAT and muscle in db/db mice. PGC-1α upregulation apparently modulated transcriptional reprogramming to increase mitochondrial mass and fatty acid oxidative metabolism in muscle, beiging of WAT, and raised circulating adiponectin level in db/db mice. This was in sharp contrast to the liver, where TNKS inhibition in db/db mice had no effect on PGC-1α expression, lipid metabolism, or gluconeogenesis. CONCLUSION: Our study unravels a novel molecular mechanism whereby pharmacological inhibition of TNKS in obesity and diabetes enhances oxidative metabolism and ameliorates lipid disorder. This happens via tissue-specific PGC-1α-driven transcriptional reprogramming in muscle and WAT, without affecting liver. This highlights inhibition of TNKS as a potential pharmacotherapy for obesity and T2DM.


Asunto(s)
Diabetes Mellitus Tipo 2/metabolismo , Dislipidemias/tratamiento farmacológico , Obesidad/metabolismo , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/metabolismo , Tanquirasas/antagonistas & inhibidores , Grasa Abdominal , Tejido Adiposo Blanco , Animales , Peso Corporal , Hígado , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos , Oxidación-Reducción , Poli ADP Ribosilación , Sulfonas/uso terapéutico , Tanquirasas/metabolismo , Triazoles/uso terapéutico
5.
J Psychol ; 154(5): 325-345, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32281919

RESUMEN

The advancement of technology has led to an increasingly permeable boundary between work and off-work time. As such, employees may face pressure to immediately respond to work-related information and communication technology (ICT) messages during off-work time. This study examines the mediating role of workplace telepressure on the relationships between ICT availability demands with burnout and work-family conflict, as well as the moderating effects of self-regulation on these relationships. Data were collected from 185 full-time employees at two time points. Results indicated full support for the moderated mediation model, demonstrating that workplace telepressure mediated the relationships between ICT availability demands and burnout and work-family conflict. Moreover, dispositional self-regulation strengthened the direct effect of ICT availability demands on workplace telepressure and the indirect effects of ICT availability demands on burnout and work-family conflict. Theoretical and practical implications are discussed.


Asunto(s)
Agotamiento Profesional , Familia , Personalidad , Autocontrol , Telecomunicaciones , Equilibrio entre Vida Personal y Laboral , Lugar de Trabajo/psicología , Niño , Crianza del Niño , Empleo , Femenino , Humanos , Masculino , Encuestas y Cuestionarios
6.
J Appl Psychol ; 105(6): 597-618, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-31556628

RESUMEN

Emotional labor, or regulating emotions as part of one's work role, is needed for performance yet may come with far-reaching costs to employee health and performance. Based on ego depletion theorizing, we propose that on days employees perform more surface acting (i.e., faking positive and hiding negative emotional expressions), they will consume more alcohol later-due to reduced self-control (i.e., depletion). In 2 studies, public-facing employees completed multiple assessments per day for 2 weeks. Study 1 showed that surface acting had no direct or indirect effect on alcohol use via depletion, nor via negative mood as an alternative measure of depletion. Study 2 demonstrated that surface acting directly increased subsequent drinking only for those with high emotional demands, but not through depletion. Across both studies, daily deep acting (i.e., modifying emotions to feel positive) consistently predicted less alcohol consumption, but this did not occur through depletion. Study 2 provided evidence for an alternative, motivational shift explanation-a reduced motive to detach from work after regulating by deep acting-rather than self-control capacity. These findings contribute to debate on ego depletion theory by providing insightful field evidence, while demonstrating when emotional labor is likely to help or harm employees' health. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Asunto(s)
Alcoholismo/psicología , Regulación Emocional , Satisfacción en el Trabajo , Estrés Laboral/psicología , Adulto , Femenino , Humanos , Masculino , Ajuste Social
7.
Sci Rep ; 9(1): 4710, 2019 03 18.
Artículo en Inglés | MEDLINE | ID: mdl-30886164

RESUMEN

Key to whole body glucose homeostasis is the ability of fat and muscle cells to sequester the facilitative glucose transporter GLUT4 in an intracellular compartment from where it can be mobilized in response to insulin. We have previously demonstrated that this process requires ubiquitination of GLUT4 while numerous other studies have identified several molecules that are also required, including the insulin-responsive aminopeptidase IRAP and its binding partner, the scaffolding protein tankyrase. In addition to binding IRAP, Tankyrase has also been shown to bind the deubiquinating enzyme USP25. Here we demonstrate that USP25 and Tankyrase interact, and colocalise with GLUT4 in insulin-sensitive cells. Furthermore depletion of USP25 from adipocytes reduces cellular levels of GLUT4 and concomitantly blunts the ability of insulin to stimulate glucose transport. Collectively, these data support our model that sorting of GLUT4 into its insulin-sensitive store involves a cycle of ubiquitination and subsequent deubiquitination.


Asunto(s)
Adipocitos/metabolismo , Cistinil Aminopeptidasa/metabolismo , Transportador de Glucosa de Tipo 4/metabolismo , Tanquirasas/metabolismo , Ubiquitina Tiolesterasa/metabolismo , Células 3T3-L1 , Adipocitos/citología , Animales , Membrana Celular/metabolismo , Técnicas de Silenciamiento del Gen , Glucosa/metabolismo , Insulina/metabolismo , Ratones , Ubiquitina Tiolesterasa/genética , Ubiquitinación
8.
Adv Mater ; 30(22): e1705605, 2018 May.
Artículo en Inglés | MEDLINE | ID: mdl-29665153

RESUMEN

Treatment with exogenous nitric oxide (NO) donors is regarded as being effective against osteoporosis. However, NO has a short half-life, limiting its clinical usefulness. To overcome this limitation, an injectable microparticle (MP) system is developed that consists of phase-change materials capric acid (CA) and octadecane, and encapsulates a NO donor. The therapeutic efficacy of the MPs is evaluated in ovariectomized (OVX) rats with osteoporosis. Upon subcutaneous administration, the MPs undergo a phase transition, leaching out the NO donor and generating NO bubbles that are instantly covered by a layer of tightly packed CA surfactant molecules, forming micellar depots. The in situ self-assembling micellar depots can actively protect the NO bubbles, prolonging their half-life, while the entrapped NO may passively diffuse through the micellar depots over time, performing a long-lasting therapeutic function, reversing the OVX-induced osteoporosis. It is possible to use the concept of in situ self-assembling micellar depots developed herein to expand the therapeutic effect of NO in its diverse range of clinical applications.


Asunto(s)
Óxido Nítrico/química , Animales , Micelas , Osteoporosis , Ratas
9.
Diabetes ; 67(5): 841-848, 2018 05.
Artículo en Inglés | MEDLINE | ID: mdl-29432123

RESUMEN

The activation of Kupffer cells (KCs) and monocyte-derived recruited macrophages (McMΦs) in the liver contributes to obesity-induced insulin resistance and type 2 diabetes. Mice with diet-induced obesity (DIO mice) treated with chromogranin A peptide catestatin (CST) showed several positive results. These included decreased hepatic/plasma lipids and plasma insulin, diminished expression of gluconeogenic genes, attenuated expression of proinflammatory genes, increased expression of anti-inflammatory genes in McMΦs, and inhibition of the infiltration of McMΦs resulting in improvement of insulin sensitivity. Systemic CST knockout (CST-KO) mice on normal chow diet (NCD) ate more food, gained weight, and displayed elevated blood glucose and insulin levels. Supplementation of CST normalized glucose and insulin levels. To verify that the CST deficiency caused macrophages to be very proinflammatory in CST-KO NCD mice and produced glucose intolerance, we tested the effects of (sorted with FACS) F4/80+Ly6C- cells (representing KCs) and F4/80-Ly6C+ cells (representing McMΦs) on hepatic glucose production (HGP). Both basal HGP and glucagon-induced HGP were markedly increased in hepatocytes cocultured with KCs and McMΦs from NCD-fed CST-KO mice, and the effect was abrogated upon pretreatment of CST-KO macrophages with CST. Thus, we provide a novel mechanism of HGP suppression through CST-mediated inhibition of macrophage infiltration and function.


Asunto(s)
Cromogranina A/farmacología , Glucosa/metabolismo , Resistencia a la Insulina , Macrófagos del Hígado/efectos de los fármacos , Hígado/efectos de los fármacos , Macrófagos/efectos de los fármacos , Obesidad/metabolismo , Fragmentos de Péptidos/farmacología , Animales , Cromogranina A/genética , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/inmunología , Glucagón/farmacología , Gluconeogénesis/efectos de los fármacos , Gluconeogénesis/genética , Hormonas/farmacología , Inflamación/inmunología , Insulina/metabolismo , Macrófagos del Hígado/inmunología , Metabolismo de los Lípidos/efectos de los fármacos , Hígado/inmunología , Hígado/metabolismo , Macrófagos/inmunología , Masculino , Ratones , Ratones Noqueados , Obesidad/inmunología , Fragmentos de Péptidos/genética
10.
J Biol Chem ; 293(13): 4735-4751, 2018 03 30.
Artículo en Inglés | MEDLINE | ID: mdl-29378845

RESUMEN

Estrogen receptor α (ERα) action plays an important role in pancreatic ß-cell function and survival; thus, it is considered a potential therapeutic target for the treatment of type 2 diabetes in women. However, the mechanisms underlying the protective effects of ERα remain unclear. Because ERα regulates mitochondrial metabolism in other cell types, we hypothesized that ERα may act to preserve insulin secretion and promote ß-cell survival by regulating mitochondrial-endoplasmic reticulum (EndoRetic) function. We tested this hypothesis using pancreatic islet-specific ERα knockout (PERαKO) mice and Min6 ß-cells in culture with Esr1 knockdown (KD). We found that Esr1-KD promoted reactive oxygen species production that associated with reduced fission/fusion dynamics and impaired mitophagy. Electron microscopy showed mitochondrial enlargement and a pro-fusion phenotype. Mitochondrial cristae and endoplasmic reticulum were dilated in Esr1-KD compared with ERα replete Min6 ß-cells. Increased expression of Oma1 and Chop was paralleled by increased oxygen consumption and apoptosis susceptibility in ERα-KD cells. In contrast, ERα overexpression and ligand activation reduced both Chop and Oma1 expression, likely by ERα binding to consensus estrogen-response element sites in the Oma1 and Chop promoters. Together, our findings suggest that ERα promotes ß-cell survival and insulin secretion through maintenance of mitochondrial fission/fusion-mitophagy dynamics and EndoRetic function, in part by Oma1 and Chop repression.


Asunto(s)
Apoptosis , Estrés del Retículo Endoplásmico , Receptor alfa de Estrógeno/metabolismo , Células Secretoras de Insulina/metabolismo , Mitocondrias/metabolismo , Mitofagia , Animales , Supervivencia Celular , Receptor alfa de Estrógeno/genética , Femenino , Insulina/genética , Insulina/metabolismo , Metaloproteasas/biosíntesis , Metaloproteasas/genética , Ratones , Ratones Noqueados , Mitocondrias/genética , Proteínas Mitocondriales/biosíntesis , Proteínas Mitocondriales/genética , Especies Reactivas de Oxígeno/metabolismo , Factor de Transcripción CHOP/biosíntesis , Factor de Transcripción CHOP/genética
11.
J Occup Health Psychol ; 23(1): 58-70, 2018 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-27786506

RESUMEN

Drawing on the stressor-emotion model, we examine how customer mistreatment can evoke service workers' passive forms of deviant behaviors (i.e., work withdrawal behavior [WWB]) and negative impacts on their home life (i.e., work-family conflict [WFC]), and whether individuals' core self-evaluations and customer service training can buffer the negative effects of customer mistreatment. Using the experience sampling method, we collect daily data from 77 customer service employees for 10 consecutive working days, yielding 546 valid daily responses. The results show that daily customer mistreatment increases service workers' daily WWB and WFC through negative emotions. Furthermore, employees with high core self-evaluations and employees who received customer service training are less likely to experience negative emotions when faced with customer mistreatment, and thus are less likely to engage in WWB or provoke WFC. (PsycINFO Database Record


Asunto(s)
Comportamiento del Consumidor , Conflicto Familiar/psicología , Relaciones Interpersonales , Negativismo , Estrés Psicológico , Trabajo/psicología , Adulto , Agresión/psicología , Femenino , Humanos , Industrias , Masculino , Persona de Mediana Edad , Ocupaciones , Autoimagen , Taiwán
12.
Cell Tissue Res ; 368(3): 487-501, 2017 06.
Artículo en Inglés | MEDLINE | ID: mdl-28220294

RESUMEN

Chromogranin A (CgA) is a prohormone and a granulogenic factor that regulates secretory pathways in neuroendocrine tissues. In ß-cells of the endocrine pancreas, CgA is a major cargo in insulin secretory vesicles. The impact of CgA deficiency on the formation and exocytosis of insulin vesicles is yet to be investigated. In addition, no literature exists on the impact of CgA on mitochondrial function in ß-cells. Using three different antibodies, we demonstrate that CgA is processed to vasostatin- and catestatin-containing fragments in pancreatic islet cells. CgA deficiency in Chga-KO islets leads to compensatory overexpression of chromogranin B, secretogranin II, SNARE proteins and insulin genes, as well as increased insulin protein content. Ultrastructural studies of pancreatic islets revealed that Chga-KO ß-cells contain fewer immature secretory granules than wild-type (WT) control but increased numbers of mature secretory granules and plasma membrane-docked vesicles. Compared to WT control, CgA-deficient ß-cells exhibited increases in mitochondrial volume, numerical densities and fusion, as well as increased expression of nuclear encoded genes (Ndufa9, Ndufs8, Cyc1 and Atp5o). These changes in secretory vesicles and the mitochondria likely contribute to the increased glucose-stimulated insulin secretion observed in Chga-KO mice. We conclude that CgA is an important regulator for coordination of mitochondrial dynamics, secretory vesicular quanta and GSIS for optimal secretory functioning of ß-cells, suggesting a strong, CgA-dependent positive link between mitochondrial fusion and GSIS.


Asunto(s)
Cromogranina A/fisiología , Insulina/metabolismo , Islotes Pancreáticos/metabolismo , Dinámicas Mitocondriales , Animales , Calreticulina/metabolismo , Diferenciación Celular , Cromogranina A/deficiencia , Cromogranina A/metabolismo , Exocitosis , Regulación de la Expresión Génica , Glucosa/metabolismo , Secreción de Insulina , Islotes Pancreáticos/citología , Islotes Pancreáticos/ultraestructura , Masculino , Ratones , Ratones Endogámicos C57BL , Dinámicas Mitocondriales/genética , Fragmentos de Péptidos/metabolismo , Vesículas Secretoras
13.
J Endocrinol ; 232(2): 137-153, 2017 02.
Artículo en Inglés | MEDLINE | ID: mdl-27799464

RESUMEN

Chromogranin A (CgA) is widely expressed in endocrine and neuroendocrine tissues as well as in the central nervous system. We observed CgA expression (mRNA and protein) in the gastrocnemius (GAS) muscle and found that performance of CgA-deficient Chga-KO mice in treadmill exercise was impaired. Supplementation with CgA in Chga-KO mice restored exercise ability suggesting a novel role for endogenous CgA in skeletal muscle function. Chga-KO mice display (i) lack of exercise-induced stimulation of pAKT, pTBC1D1 and phospho-p38 kinase signaling, (ii) loss of GAS muscle mass, (iii) extensive formation of tubular aggregates (TA), (iv) disorganized cristae architecture in mitochondria, (v) increased expression of the inflammatory cytokines Tnfα, Il6 and Ifnγ, and fibrosis. The impaired maximum running speed and endurance in the treadmill exercise in Chga-KO mice correlated with decreased glucose uptake and glycolysis, defects in glucose oxidation and decreased mitochondrial cytochrome C oxidase activity. The lack of adaptation to endurance training correlated with the lack of stimulation of p38MAPK that is known to mediate the response to tissue damage. As CgA sorts proteins to the regulated secretory pathway, we speculate that lack of CgA could cause misfolding of membrane proteins inducing aggregation of sarcoplasmic reticulum (SR) membranes and formation of tubular aggregates that is observed in Chga-KO mice. In conclusion, CgA deficiency renders the muscle energy deficient, impairs performance in treadmill exercise and prevents regeneration after exercise-induced tissue damage.


Asunto(s)
Cromogranina A/metabolismo , Resistencia a la Insulina/fisiología , Músculo Esquelético/metabolismo , Condicionamiento Físico Animal/fisiología , Animales , Cromogranina A/genética , Cromogranina A/farmacología , Complejo IV de Transporte de Electrones/metabolismo , Glucosa/metabolismo , Glucólisis/fisiología , Ratones , Ratones Noqueados , Mitocondrias/metabolismo , Músculo Esquelético/efectos de los fármacos , Fosforilación/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo
14.
J Bone Miner Res ; 31(10): 1841-1844, 2016 10.
Artículo en Inglés | MEDLINE | ID: mdl-27105398

RESUMEN

The CYP24A1 gene encodes a mitochondrial 24-hydroxylase that inactivates 1,25(OH)2 D. Loss-of-function mutations in CYP24A1 cause hypercalcemia, nephrolithiasis and nephrocalcinosis. We describe a woman with CYP24A1 deficiency and recurrent gestational hypercalcemia. Her first pregnancy, at age 20, resulted with the intrauterine demise of twin fetuses. Postpartum, she developed severe hypercalcemia (14 mg/dL), altered mental status, and acute pancreatitis. Her PTH was suppressed (6 pg/mL) and her 1,25(OH)2 D was elevated (165 and 195 pg/mL on postpartum day 1 and 5, respectively). Between one and three months postpartum, her serum calcium decreased from 11.4 to 10.2 mg/dL while her 1,25(OH)2 D level decreased from 83 to 24 pg/mL. Her 24-hour urine calcium was 277 mg. Six months postpartum, she became pregnant again. At 14 weeks, her albumin-corrected calcium level was 10.4 mg/dL and her 1,25(OH)2 D level exceeded 200 pg/mL. To establish the diagnosis of CYP24A1 deficiency, we showed her 24,25(OH)2 D level to be undetectable (<2 ng/mL). Exon sequencing of the CYP24A1 gene revealed a homozygous, 8-nucleotide deletion in exon 8, causing an S334V substitution and premature termination due to a frame shift (c.999_1006del, p.Ser334Valfs*9). To prevent hypercalcemia, she was advised to discontinue prenatal vitamins, avoid sun exposure and calcium-rich foods, and start omeprazole and a calcium binder (250 mg K-Phos-neutral with meals). Despite these measures, both hypercalcemia (11.5 mg/dL) and acute pancreatitis recurred. Labor was induced and a healthy, normocalcemic boy was delivered. In the absence of lactation, maternal hypercalcemia resolved within 2 months. This report shows that CYP24A1-deficient subjects may be normocalcemic at baseline. Hypercalcemia may be unmasked by pregnancy through the routine use of calciferol-containing prenatal vitamins, increased 1-alpha hydroxylation of VitD by the placenta and maternal kidney, and production of PTHrP by the uteroplacental unit. CYP24A1 deficiency should be considered in patients with unexplained vitamin D-mediated hypercalcemia. © 2016 American Society for Bone and Mineral Research.


Asunto(s)
Secuencia de Bases , Hipercalcemia , Pancreatitis , Trastornos Puerperales , Eliminación de Secuencia , Vitamina D3 24-Hidroxilasa/deficiencia , Enfermedad Aguda , Adulto , Femenino , Humanos , Hipercalcemia/sangre , Hipercalcemia/tratamiento farmacológico , Hipercalcemia/genética , Pancreatitis/sangre , Pancreatitis/tratamiento farmacológico , Pancreatitis/genética , Embarazo , Trastornos Puerperales/sangre , Trastornos Puerperales/tratamiento farmacológico , Trastornos Puerperales/genética
15.
Am J Physiol Endocrinol Metab ; 310(11): E874-85, 2016 06 01.
Artículo en Inglés | MEDLINE | ID: mdl-27072493

RESUMEN

Contact between ß-cells is necessary for their normal function. Identification of the proteins mediating the effects of ß-cell-to-ß-cell contact is a necessary step toward gaining a full understanding of the determinants of ß-cell function and insulin secretion. The secretory machinery of the ß-cells is nearly identical to that of central nervous system (CNS) synapses, and we hypothesize that the transcellular protein interactions that drive maturation of the two secretory machineries upon contact of one cell (or neural process) with another are also highly similar. Two such transcellular interactions, important for both synaptic and ß-cell function, have been identified: EphA/ephrin-A and neuroligin/neurexin. Here, we tested the role of another synaptic cleft protein, CADM1, in insulinoma cells and in rat and human islet ß-cells. We found that CADM1 is a predominant CADM isoform in ß-cells. In INS-1 cells and primary ß-cells, CADM1 constrains insulin secretion, and its expression decreases after prolonged glucose stimulation. Using a coculture model, we found that CADM1 also influences insulin secretion in a transcellular manner. We asked whether extracellular CADM1 interactions exert their influence via the same mechanisms by which they influence neurotransmitter exocytosis. Our results suggest that, as in the CNS, CADM1 interactions drive exocytic site assembly and promote actin network formation. These results support the broader hypothesis that the effects of cell-cell contact on ß-cell maturation and function are mediated by the same extracellular protein interactions that drive the formation of the presynaptic exocytic machinery. These interactions may be therapeutic targets for reversing ß-cell dysfunction in diabetes.


Asunto(s)
Moléculas de Adhesión Celular/metabolismo , Exocitosis/fisiología , Inmunoglobulinas/metabolismo , Insulina/metabolismo , Islotes Pancreáticos/metabolismo , Sintaxina 1/metabolismo , Animales , Molécula 1 de Adhesión Celular , Comunicación Celular/fisiología , Línea Celular , Líquido Extracelular/metabolismo , Humanos , Secreción de Insulina , Ratas , Especificidad de la Especie
16.
Diabetologia ; 59(3): 582-91, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26631215

RESUMEN

AIMS/HYPOTHESIS: Tankyrase (TNKS) is a ubiquitously expressed molecular scaffold that is implicated in diverse processes. The catalytic activity of TNKS modifies substrate proteins through poly-ADP-ribosylation (PARsylation) and is responsive to cellular energetic state. Global deficiency of the TNKS protein in mice accelerates glucose utilisation and raises plasma adiponectin levels. The aim of this study was to investigate whether the PARsylation activity of TNKS in adipocytes plays a role in systemic glucose homeostasis. METHODS: To inhibit TNKS-mediated PARsylation, we fed mice with a diet containing the TNKS-specific inhibitor G007-LK. To genetically inactivate TNKS catalysis in adipocytes while preserving its function as a molecular scaffold, we used an adipocyte-selective Cre transgene to delete TNKS exons that encoded the catalytic domain at the C-terminus. Tissue-specific insulin sensitivity in mice was investigated using hyperinsulinaemic-euglycaemic clamps. To model adipose-liver crosstalk ex vivo, we applied adipocyte-conditioned media to hepatocytes and assessed the effect on gluconeogenesis. RESULTS: The TNKS inhibitor G007-LK improved glucose tolerance and insulin sensitivity and promptly increased plasma adiponectin levels. In female mice, but not in male mice, adipocyte-selective genetic inactivation of TNKS catalysis improved hepatic insulin sensitivity and post-transcriptionally increased plasma adiponectin levels. Both pharmacological and genetic TNKS inhibition in female mouse-derived adipocytes induced a change in secreted factors to decrease gluconeogenesis in primary hepatocytes. CONCLUSIONS/INTERPRETATION: Systemic glucose homeostasis is regulated by the PARsylation activity of TNKS in adipocytes. This regulation is mediated in part by adipocyte-secreted factors that modulate hepatic glucose production. Pharmacological TNKS inhibition could potentially be used to improve glucose tolerance.


Asunto(s)
Tejido Adiposo/efectos de los fármacos , Tejido Adiposo/enzimología , Glucosa/metabolismo , Tanquirasas/metabolismo , Animales , Glucemia/efectos de los fármacos , Metabolismo de los Hidratos de Carbono/efectos de los fármacos , Femenino , Masculino , Ratones , Sulfonas/farmacología , Tanquirasas/antagonistas & inhibidores , Triazoles/farmacología
17.
Am J Trop Med Hyg ; 93(4): 822-7, 2015 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-26195463

RESUMEN

A 50-year-old male immigrant from Ethiopia presented for consultation after 3 years of hematochezia/melena requiring > 25 units of blood transfusions. Physical examination revealed severe proximal muscle wasting and weakness, central obesity, proptosis, and abdominal striae, accompanied by eosinophilia, elevated hemoglobin A1c, elevated 24-hour urinary cortisol, lack of suppression of 8 am cortisol levels by 1 mg dexamethasone, and inappropriately elevated random adrenocorticotropic hormone (ACTH) level. Histopathological examination of gastrointestinal biopsies showed large numbers of Strongyloides stercoralis, indicating Strongyloides hyperinfection. Treatment with 2 days of ivermectin led to resolution of gastrointestinal bleeding. This syndrome was due to chronic immunosuppression from a pituitary ACTH (corticotroph) microadenoma, of which resection led to gradual normalization of urine cortisol, improved glycemic control, resolution of eosinophilia, and no recurrence of infection.


Asunto(s)
Hemorragia Gastrointestinal/etiología , Hipersecreción de la Hormona Adrenocorticotrópica Pituitaria (HACT)/diagnóstico , Strongyloides stercoralis , Estrongiloidiasis/diagnóstico , Enfermedad Aguda , Animales , Antihelmínticos/uso terapéutico , Hemorragia Gastrointestinal/parasitología , Humanos , Huésped Inmunocomprometido/inmunología , Ivermectina/uso terapéutico , Masculino , Persona de Mediana Edad , Hipersecreción de la Hormona Adrenocorticotrópica Pituitaria (HACT)/complicaciones , Hipersecreción de la Hormona Adrenocorticotrópica Pituitaria (HACT)/inmunología , Hipersecreción de la Hormona Adrenocorticotrópica Pituitaria (HACT)/parasitología , Hipersecreción de la Hormona Adrenocorticotrópica Pituitaria (HACT)/patología , Adenohipófisis/patología , Estrongiloidiasis/complicaciones
18.
J Biol Chem ; 290(23): 14454-61, 2015 Jun 05.
Artículo en Inglés | MEDLINE | ID: mdl-25944897

RESUMEN

In adipose and muscle cells, insulin stimulates the exocytic translocation of vesicles containing GLUT4, a glucose transporter, and insulin-regulated aminopeptidase (IRAP), a transmembrane aminopeptidase. A substrate of IRAP is vasopressin, which controls water homeostasis. The physiological importance of IRAP translocation to inactivate vasopressin remains uncertain. We previously showed that in skeletal muscle, insulin stimulates proteolytic processing of the GLUT4 retention protein, TUG, to promote GLUT4 translocation and glucose uptake. Here we show that TUG proteolysis also controls IRAP targeting and regulates vasopressin action in vivo. Transgenic mice with constitutive TUG proteolysis in muscle consumed much more water than wild-type control mice. The transgenic mice lost more body weight during water restriction, and the abundance of renal AQP2 water channels was reduced, implying that vasopressin activity is decreased. To compensate for accelerated vasopressin degradation, vasopressin secretion was increased, as assessed by the cosecreted protein copeptin. IRAP abundance was increased in T-tubule fractions of fasting transgenic mice, when compared with controls. Recombinant IRAP bound to TUG, and this interaction was mapped to a short peptide in IRAP that was previously shown to be critical for GLUT4 intracellular retention. In cultured 3T3-L1 adipocytes, IRAP was present in TUG-bound membranes and was released by insulin stimulation. Together with previous results, these data support a model in which TUG controls vesicle translocation by interacting with IRAP as well as GLUT4. Furthermore, the effect of IRAP to reduce vasopressin activity is a physiologically important consequence of vesicle translocation, which is coordinated with the stimulation of glucose uptake.


Asunto(s)
Proteínas Portadoras/metabolismo , Glucosa/metabolismo , Músculo Esquelético/metabolismo , Vasopresinas/metabolismo , Células 3T3-L1 , Animales , Transporte Biológico , Cistinil Aminopeptidasa/metabolismo , Exocitosis , Transportador de Glucosa de Tipo 4/metabolismo , Insulina/metabolismo , Péptidos y Proteínas de Señalización Intracelular , Ratones , Ratones Endogámicos C57BL
19.
PLoS One ; 10(4): e0122948, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25876076

RESUMEN

The poly-ADP-ribosylation (PARsylation) activity of tankyrase (TNKS) regulates diverse physiological processes including energy metabolism and wnt/ß-catenin signaling. This TNKS activity uses NAD+ as a co-substrate to post-translationally modify various acceptor proteins including TNKS itself. PARsylation by TNKS often tags the acceptors for ubiquitination and proteasomal degradation. Whether this TNKS activity is regulated by physiological changes in NAD+ levels or, more broadly, in cellular energy charge has not been investigated. Because the NAD+ biosynthetic enzyme nicotinamide phosphoribosyltransferase (NAMPT) in vitro is robustly potentiated by ATP, we hypothesized that nutritional energy might stimulate cellular NAMPT to produce NAD+ and thereby augment TNKS catalysis. Using insulin-secreting cells as a model, we showed that glucose indeed stimulates the autoPARsylation of TNKS and consequently its turnover by the ubiquitin-proteasomal system. This glucose effect on TNKS is mediated primarily by NAD+ since it is mirrored by the NAD+ precursor nicotinamide mononucleotide (NMN), and is blunted by the NAMPT inhibitor FK866. The TNKS-destabilizing effect of glucose is shared by other metabolic fuels including pyruvate and amino acids. NAD+ flux analysis showed that glucose and nutrients, by increasing ATP, stimulate NAMPT-mediated NAD+ production to expand NAD+ stores. Collectively our data uncover a metabolic pathway whereby nutritional energy augments NAD+ production to drive the PARsylating activity of TNKS, leading to autoPARsylation-dependent degradation of the TNKS protein. The modulation of TNKS catalytic activity and protein abundance by cellular energy charge could potentially impose a nutritional control on the many processes that TNKS regulates through PARsylation. More broadly, the stimulation of NAD+ production by ATP suggests that nutritional energy may enhance the functions of other NAD+-driven enzymes including sirtuins.


Asunto(s)
Insulinoma/patología , NAD/química , Tanquirasas/química , Células 3T3 , Acrilamidas/química , Adenosina Trifosfato/química , Animales , Catálisis , Metabolismo Energético/genética , Glucosa/química , Células HEK293 , Humanos , Ratones , Nicotinamida Fosforribosiltransferasa/química , Piperidinas/química , Complejo de la Endopetidasa Proteasomal/química , Procesamiento Proteico-Postraduccional , Ratas , Ubiquitina/química
20.
Diabetes ; 64(1): 104-16, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25048197

RESUMEN

Chromogranin A knockout (Chga-KO) mice exhibit enhanced insulin sensitivity despite obesity. Here, we probed the role of the chromogranin A-derived peptide pancreastatin (PST: CHGA(273-301)) by investigating the effect of diet-induced obesity (DIO) on insulin sensitivity of these mice. We found that on a high-fat diet (HFD), Chga-KO mice (KO-DIO) remain more insulin sensitive than wild-type DIO (WT-DIO) mice. Concomitant with this phenotype is enhanced Akt and AMPK signaling in muscle and white adipose tissue (WAT) as well as increased FoxO1 phosphorylation and expression of mature Srebp-1c in liver and downregulation of the hepatic gluconeogenic genes, Pepck and G6pase. KO-DIO mice also exhibited downregulation of cytokines and proinflammatory genes and upregulation of anti-inflammatory genes in WAT, and peritoneal macrophages from KO mice displayed similarly reduced proinflammatory gene expression. The insulin-sensitive, anti-inflammatory phenotype of KO-DIO mice is masked by supplementing PST. Conversely, a PST variant peptide PSTv1 (PST-NΔ3: CHGA(276-301)), lacking PST activity, simulated the KO phenotype by sensitizing WT-DIO mice to insulin. In summary, the reduced inflammation due to PST deficiency prevented the development of insulin resistance in KO-DIO mice. Thus, obesity manifests insulin resistance only in the presence of PST, and in its absence obesity is dissociated from insulin resistance.


Asunto(s)
Cromogranina A/inmunología , Obesidad/inmunología , Obesidad/metabolismo , Hormonas Pancreáticas/farmacología , Paniculitis/inmunología , Transducción de Señal/inmunología , Tejido Adiposo/inmunología , Tejido Adiposo/metabolismo , Animales , Células Cultivadas , Quimiotaxis/inmunología , Cromogranina A/genética , Cromogranina A/metabolismo , Proteína Forkhead Box O1 , Factores de Transcripción Forkhead/inmunología , Factores de Transcripción Forkhead/metabolismo , Intolerancia a la Glucosa/tratamiento farmacológico , Intolerancia a la Glucosa/inmunología , Intolerancia a la Glucosa/metabolismo , Resistencia a la Insulina/inmunología , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/inmunología , Macrófagos Peritoneales/metabolismo , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Obesidad/tratamiento farmacológico , Hormonas Pancreáticas/inmunología , Hormonas Pancreáticas/metabolismo , Paniculitis/tratamiento farmacológico , Paniculitis/metabolismo , Transducción de Señal/efectos de los fármacos , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/inmunología , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/metabolismo
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