Proliferación de células β pancreáticas en ratones diabéticos tratados con Cucurbita ficifolia Bouché

En México, la diabetes mellitus (DM) es una de las principales causas de muerte debido a las complicaciones vasculares que la acompañan, reduciendo la expectativa de vida de las personas que la padecen y ubicándose como una enfermedad con gastos excesivos en su prevención y control. Se estima que en el mundo hay más de 346 millones de adultos con diabetes y que en el 2030 se convertirá en la séptima causa de muerte. La DM caracteriza por un estado de hiperglucemia crónico debido a defectos en la acción y/o secreción de insulina, además de presentar alteraciones en el metabolismo de carbohidratos, lípidos y proteínas. La diabetes tipo 2 (DT2) se presenta principalmente en edad adulta y se caracteriza por resistencia y disminución de la insulina, teniendo como consecuencia la disfunción progresiva de las células β del páncreas, las cuales tienen una baja capacidad de regeneración; sin embargo, hoy en día se tienen evidencias de la existencia de marcadores moleculares que regulan el ciclo de la célula β, como la cinasa tipo 4 dependiente de ciclina (CDK4), involucrada en la capacidad de regeneración, crecimiento y con la vía de síntesis de insulina, ya que se encuentra regulando la fase G1 de ciclo celular, representando uno de los marcadores más importantes de proliferación de la célula β. La importancia epidemiológica de la diabetes y el desarrollo de las complicaciones vasculares ha llevado a proponer nuevas alternativas farmacológicas que disminuyan dichas complicaciones. En México, una planta que se utiliza empíricamente en el control de la DT2 es Cucurbita ficifolia Bouché (C. ficifolia). Se tienen evidencias experimentales y clínicas acerca de su acción hipoglucemiante. Así mismo, algunas observaciones histológicas preliminares sugieren que C. ficifolia podría aumentar la densidad celular en los islotes de Langerhans del páncreas de ratón. Sin embargo, dichas observaciones requieren ser confirmadas a través de estudios morfológicos cuantitativos e inmunohistoquímicos más precisos. En el presente estudio se realizó un estudio morfológico en hígado, riñón y páncreas de ratones sanos y diabéticos tratados diariamente con C. ficifolia, cuantificando en este último órgano el número de islotes y su densidad celular. Además, se estudió el efecto sobre CDK4 (un marcador de proliferación celular), así como sobre las concentraciones séricas de insulina en el mismo modelo. Esto con la finalidad de conocer si C. ficifolia es capaz de incrementar la producción de insulina por un mecanismo que podría implicar regeneración de las células β pancreáticas. Los resultados mostraron que la administración de C. ficifolia a ratones sanos y diabéticos reduce los niveles de glucemia y corrige algunos parámetros característicos de DT2. Además, C. ficifolia incrementó las concentraciones de insulina, así como de CDK4 en los islotes pancreáticos, sin alterar la morfología de este órgano. C ficifolia tendió a mantener la estructura general del hígado y riñón. C. ficifolia modificó también la distribución del glucógeno hepático, sugiriendo que el mecanismo de acción de la planta podría estar mediado tanto por la síntesis de insulina a través de CDK4, como por la vía de síntesis de glucógeno. C. ficifolia representa una alternativa terapéutica para el tratamiento de la diabetes mellitus, con efecto hipoglucémico en hígado y regenerador pancreático, lo que potencialmente podría ayudar a un mejor control de la diabetes, previniendo el desarrollo de complicaciones vasculares y mejorando la calidad de vida de los pacientes.In Mexico, diabetes mellitus (DM) is one of the leading causes of death due to vascular complications that accompany it, reducing the life expectancy of the patients, and it is considered as a disease with excessive cost for prevention and control. It is estimated that the world has more than 346 million people with diabetes and in 2030 will become the seventh leading cause of death. The DM is characterized by a chronic state of hyperglycemia due to defects in the action and/or secretion of insulin, causing alterations in the metabolism of carbohydrates, lipids and proteins. In particular, type 2 diabetes (DT2) occurs mostly in adulthood and is characterized by resistance and decreased in the secretion of insulin, as resulted of progressive dysfunction of pancreatic β cells, which have a low capacity for regeneration. The cyclin-dependent kinase type 4 (CDK4), which is involved in the cell regeneration capacity, as well as in the growth and biosynthesis of insulin, which regulating cell cycle in the G1 phase, represents one of the most important markers of proliferation of the β cell. The high incidence, prevalence and mortality due to diabetes, as well as its vascular complications, have led to propose new pharmacological alternatives of treatment. Cucurbita ficifolia Bouché (C. ficifolia), a plant used empirically in the control of DT2 in Mexico, has shown hypoglycemic activity in clinical and experimental studies. In addition, some preliminary histological observations suggest the possibility of increased cell-β density in the islets of Langerhans of mouse pancreas. However, these observations require support through morphological and immuno-histochemical quantitative studies. In order to establish the basis for proposing to C.ficifolia like an alternative therapy with capacity of regenerate the pancreas, through the augmenting the proliferation of cells-β pancreatic, which may avoid the development of vascular complications, in the present research a morphological study was performed in liver, kidney and pancreas in normal and diabetic mice treated with C. ficifolia, quantifying the number of islets and its cellular density in pancreas. In addition, the effect on CDK4 (a cell proliferation marker) was evaluated, as well as on the serum concentrations of insulin in the same experimental model. The results showed that C. ficifolia reduces blood glucose levels, correcting some characteristic parameters of DT2. In addition, C. ficifolia increased insulin levels, as well as CDK4 in the pancreatic islets without altering the morphology of this organ. C. ficifolia treated of maintain the overall structure of the liver and kidney. C. ficifolia also modified the distribution of liver glycogen, suggesting that in the mechanism of action of the extract may be involved both the synthesis of insulin by CDK4 as the glycogen synthesis pathway. C. ficifolia may be considered a therapeutic alternative in the treatment of diabetes mellitus, with both liver hypoglycemic effect and pancreatic regenerator, which could slow the development of vascular complications and improve quality of life of patients with diabetes

Engineered microenvironments for synergistic VEGF - integrin signalling during vascularization

We have engineered polymer-based microenvironments that promote vasculogenesis both in vitro and in vivo through synergistic integrin-growth factor receptor signalling. Poly(ethyl acrylate) (PEA) triggers spontaneous organization of fibronectin (FN) into nanonetworks which provide availability of critical binding domains. Importantly, the growth factor binding (FNIII12-14) and integrin binding (FNIII9-10) regions are simultaneously available on FN fibrils assembled on PEA. This material platform promotes synergistic integrin/VEGF signalling which is highly effective for vascularization events in vitro with low concentrations of VEGF. VEGF specifically binds to FN fibrils on PEA compared to control polymers (poly(methyl acrylate), PMA) where FN remains in a globular conformation and integrin/GF binding domains are not simultaneously available. The vasculogenic response of human endothelial cells seeded on these synergistic interfaces (VEGF bound to FN assembled on PEA) was significantly improved compared to soluble administration of VEGF at higher doses. Early onset of VEGF signalling (PLCγ1 phosphorylation) and both integrin and VEGF signalling (ERK1/2 phosphorylation) were increased only when VEGF was bound to FN nanonetworks on PEA, while soluble VEGF did not influence early signalling. Experiments with mutant FN molecules with impaired integrin binding site (FN-RGE) confirmed the role of the integrin binding site of FN on the vasculogenic response via combined integrin/VEGF signalling. In vivo experiments using 3D scaffolds coated with FN and VEGF implanted in the murine fat pad demonstrated pro-vascularization signalling by enhanced formation of new tissue inside scaffold pores. PEA-driven organization of FN promotes efficient presentation of VEGF to promote vascularization in regenerative medicine applications

Chemical characterization of a hypoglycemic extract from Cucurbita ficifolia bouche that induces liver glycogen accumulation in diabetic mice

Background: The aqueous extract of Cucurbita ficifolia (C. ficifolia) fruit has demonstrated hypoglycemic effect, which may be attributed to some components in the extract. However, the major secondary metabolites in this fruit have not yet been identified and little is known about its extra-pancreatic action, in particular, on liver carbohydrate metabolism. Therefore, in addition to the isolation and structural elucidation of the principal components in the aqueous extract of C. ficifolia, the aim of this study was to determine whether or not the hypoglycemic effect of the aqueous extract of Cucurbita ficifolia (C. ficifolia) fruit is due to accumulation of liver glycogen in diabetic mice.Materials and Methods: The aqueous extract from fruit of C. ficifolia was fractionated and its main secondary metabolites were purified and chemically characterized (NMR and GC-MS). Alloxan-induced diabetic mice received daily by gavage the aqueous extract (30 days). The liver glycogen content was quantified by spectroscopic method and by PAS stain; ALT and AST by spectrometric method; glycogen synthase, glycogen phosphorylase and GLUT2 by Western blot; the mRNA expression of GLUT2 and glucagon-receptor by RT-PCR; while serum insulin was quantified by ELISA method. A liver histological analysis was also performed by H&E stain.Results: Chemical fingerprint showed five majoritarian compounds in the aqueous extract of C. ficifolia: p-coumaric acid, p-hydroxybenzoic acid, salicin, stigmast-7,2,2-dien-3-ol and stigmast-7-en-3-ol. The histological analysis showed accumulation of liver glycogen. Also, increased glycogen synthase and decreased glycogen phosphorylase were observed. Interestingly, the histological architecture evidenced a liver-protective effect due the extract.Conclusion: Five compounds were identified in C. ficifolia aqueous extract. The hypoglycemic effect of this extract may be partially explained by liver glycogen accumulation. The bioactive compound responsible for the hypoglycemic effect of this extract will be elucidated in subsequent studies.Keywords: Cucurbita ficifolia, Cucurbitaceae, liver glycogen, hypoglycemic plants, p-coumaric acid, salicin, p-hydroxybenzoic aci

A Rare Case of Double Belly Soleus Muscle

The rare anatomical anomaly of a double belly soleus muscle has been associated with some clinical and non-clinical correlations in patients. With symptomatic patients, usually not presenting until 20–30 years old, pain is exacerbated by long walks, running, and standing for long periods of time. This presentation could mimic the properties of a soft tissue tumor, thus misleading physicians. The discovery of the double belly soleus muscle is noticed while in surgery, biopsy, computed tomography scan, or magnetic resonance imaging machine. We herewith present a rare case of double belly soleus on the left and right posterior lower leg region of a 58-year-old female cadaver. This article will help bring awareness to the signs and symptoms of this rare anatomical anomaly

CHEMICAL CHARACTERIZATION OF A HYPOGLYCEMIC EXTRACT FROM CUCURBITA FICIFOLIA BOUCHE THAT INDUCES LIVER GLYCOGEN ACCUMULATION IN DIABETIC MICE.

Background: The aqueous extract of Cucurbita ficifolia (C. ficifolia) fruit has demonstrated hypoglycemic effect, which may be attributed to some components in the extract. However, the major secondary metabolites in this fruit have not yet been identified and little is known about its extra-pancreatic action, in particular, on liver carbohydrate metabolism. Therefore, in addition to the isolation and structural elucidation of the principal components in the aqueous extract of C. ficifolia, the aim of this study was to determine whether or not the hypoglycemic effect of the aqueous extract of Cucurbita ficifolia (C. ficifolia) fruit is due to accumulation of liver glycogen in diabetic mice. Materials and Methods: The aqueous extract from fruit of C. ficifolia was fractionated and its main secondary metabolites were purified and chemically characterized (NMR and GC-MS). Alloxan-induced diabetic mice received daily by gavage the aqueous extract (30 days). The liver glycogen content was quantified by spectroscopic method and by PAS stain; ALT and AST by spectrometric method; glycogen synthase, glycogen phosphorylase and GLUT2 by Western blot; the mRNA expression of GLUT2 and glucagon-receptor by RT-PCR; while serum insulin was quantified by ELISA method. A liver histological analysis was also performed by H&E stain. Results: Chemical fingerprint showed five majoritarian compounds in the aqueous extract of C. ficifolia: p-coumaric acid, p-hydroxybenzoic acid, salicin, stigmast-7,2,2-dien-3-ol and stigmast-7-en-3-ol. The histological analysis showed accumulation of liver glycogen. Also, increased glycogen synthase and decreased glycogen phosphorylase were observed. Interestingly, the histological architecture evidenced a liver-protective effect due the extract. Conclusion: Five compounds were identified in C. ficifolia aqueous extract. The hypoglycemic effect of this extract may be partially explained by liver glycogen accumulation. The bioactive compound responsible for the hypoglycemic effect of this extract will be elucidated in subsequent studies

Pharmacokinetics of esomeprazole in goats (Capra aegagrus hircus) after intravenous and subcutaneous administration

Background: Stressed and hospitalized goats are at risk of developing abomasal (gastric) ulceration, but there is a paucity of pharmacokinetic studies for proton pump inhibiting drugs, such as, esomeprazole in goats. Objectives: The objectives for this study were to estimate plasma pharmacokinetic parameters for esomeprazole in adult goats after intravenous (IV) and subcutaneous (SQ) administration. A secondary objective was to describe the plasma kinetics of the metabolite esomeprazole sulfone after IV and SC administration in goats. Materials and methods: Esomeprazole was administered to 5 adult goats in a crossover study at doses of 1 mg/kg IV or 2 mg/kg SC. Plasma samples were collected over 36 h and analyzed via reverse phase HPLC to determine concentrations of esomeprazole and esomeprazole sulfone. Pharmacokinetic parameters were derived via non-compartmental analysis. Results: Following IV administration, mean values for plasma clearance (Cl), elimination half-life [T1/2 (λz)], C0, and volume of distribution (Vz) of esomeprazole were estimated at 24.9 mL/min/kg, 6 min, 2.324 μg/mL, and 0.23 L/kg, respectively. After SC administration elimination half-life, maximum concentration (Cmax) and time to maximum concentration (Tmax) of esomeprazole were estimated at 29 min, 1.038 μg/mL, and 22 minutes respectively. Maximum concentrations of the sulfone metabolite were 32 and 18 ng/mL after IV and SC administration. Conclusion: Esomeprazole was rapidly eliminated from plasma after both IV and SC injection in goats. The elimination half-life in goats appears to be shorter than reported in dogs, as well as less than that reported for pantoprazole in goats. The sulfone metabolite was detected and also rapidly eliminated from the plasma after both IV and SC administration. Additional pharmacodynamic investigations are needed to determine the efficacy of esomeprazole on abomasal (gastric) acid suppression in goats and could include larger doses or additional routes of administration

Disseminated histoplasmosis and hemophagocytic syndrome in renal transplantation

El síndrome hemofagocítico es una entidad causada por hipercitoquinemia y alteraciones del sistema inmune, puede ser hereditario o adquirido y genera alta mortalidad pese al tratamiento. Dentro de las etiologías, las infecciones ocupan un lugar importante, una de ellas la histoplasmosis diseminada, infección oportunista de difícil diagnóstico que afecta a pacientes inmunosuprimidos. Presentamos el caso de una paciente, adulta mayor, trasplantada de riñón, quien presenta síndrome hemofagocítico secundario a histoplasmosis diseminada y, quien presenta respuesta adecuada con el tratamiento instaurado.Reporte de caso93-98Hemophagocytic syndrome is an entity caused by hypercytokinemia and impaired immune system, may be hereditary or acquired and generates high mortality despite treatment. Among the etiologies, infections occupy an important place, one of them disseminated histoplasmosis, an opportunistic infection of difficult diagnosis which affects immunosuppressed patients. We present the case of an adult patient, with renal transplant who developed hemophagocytic syndrome secondary to disseminated histoplasmosis and responded adequately to the antifungal treatment

Information and communication technologies for approaching smokers : a descriptive study in primary healthcare

Background: Common interventions for smoking cessation are based on medical advice and pharmacological aid. Information and communication technologies may be helpful as interventions by themselves or as complementary tools to quit smoking. The objective of the study was to determine the use of information and communication technologies (ICTs) in the smoking population attended in primary care, and describe the major factors associated with its use. Methods: Descriptive observational study in 84 health centres in Cataluña, Aragon and Salamanca. We included by simple random sampling 1725 primary healthcare smokers (any amount of tobacco) aged 18-85. Through personal interview professionals collected Socio-demographic data and variables related with tobacco consumption and ICTs use were collected through face to face interviews Factors associated with the use of ICTs were analyzed by logistic regression. Results: Users of at least one ICT were predominantly male, young (18-45 years), from most favoured social classes and of higher education. Compared with non-ICTs users, users declared lower consumption of tobacco, younger onset age, and lower nicotine dependence. The percentages of use of email, text messages and web pages were 65.3%, 74.0% and 71.5%, respectively. Factors associated with the use of ICTs were age, social class, educational level and nicotine dependence level. The factor most closely associated with the use of all three ICTs was age; mainly individuals aged 18-24. Conclusions: The use of ICTs to quit smoking is promising, with the technology of mobile phones having a broader potential. Younger and more educated subjects are good targets for ICTs interventions on smoking cessation