NGF effects promote the maturation of rat pancreatic beta cells by regulating GLUT2 levels and distribution, and glucokinase activity.

The nerve growth factor (NGF) participates in cell survival and glucose-stimulated insulin secretion (GSIS) processes in rat adult beta cells. GSIS is a complex process in which metabolic events and ionic channel activity are finely coupled. GLUT2 and glucokinase (GK) play central roles in GSIS by regulating the rate of the glycolytic pathway. The biphasic release of insulin upon glucose stimulation characterizes mature adult beta cells. On the other hand, beta cells obtained from neonatal, suckling, and weaning rats are considered immature because they secrete low levels of insulin and do not increase insulin secretion in response to high glucose. The weaning of rats (at postnatal day 20 in laboratory conditions) involves a dietary transition from maternal milk to standard chow. It is characterized by increased basal plasma glucose levels and insulin levels, which we consider physiological insulin resistance. On the other hand, we have observed that incubating rat beta cells with NGF increases GSIS by increasing calcium currents in neonatal cells. In this work, we studied the effects of NGF on the regulation of cellular distribution and activity of GLUT2 and GK to explore its potential role in the maturation of GSIS in beta cells from P20 rats. Pancreatic islet cells from both adult and P20 rats were isolated and incubated with 5.6 mM or 15.6 mM glucose with and without NGF for 4 hours. Specific immunofluorescence assays were conducted following the incubation period to detect insulin and GLUT2. Additionally, we measured glucose uptake, glucokinase activity, and insulin secretion assays at 5.6 mM or 15.6 mM glucose concentrations. We observed an age-dependent variation in the distribution of GLUT2 in pancreatic beta cells and found that glucose plays a regulatory role in GLUT2 distribution independently of age. Moreover, NGF increases GLUT2 abundance, glucose uptake, and GSIS in P20 beta cells and GK activity in adult beta cells. Our results suggest that besides increasing calcium currents, NGF regulates metabolic components of the GSIS, thereby contributing to the maturation process of pancreatic beta cells.

Inhibitory effects of methanolic extracts from the stem barks of ten Mexican Bursera Jacq. ex L. species on the activity of α-amylase and acetylcholinesterase from Tenebrio molitor.

Methanolic stem bark extracts from ten Mexican Bursera Jacq. ex L. species were evaluated in vitro with regard to their inhibitory activity against two Tenebrio molitor-derived enzymes. Seven extracts (B. bicolor, B. copallifera, B. fagaroides, B. grandifolia, B. lancifolia, B. linanoe, and B. longipes) reduced α-amylase activity by 55.37% to 96.25%, with three samples proving to be particularly potent α-amylase inhibitors (B. grandifolia, B. lancifolia, and B. linanoe; IC50 = 162, 132, and 186 µg/mL, respectively). In contrast, no extract inhibited acetylcholinesterase activity by more than 39.94%. Quantitative HPLC analysis did not reveal any clear correlation between the species-specific flavonoid or phenolic acid profiles and the respective extracts' enzyme inhibitory activity. The findings reported herein do not only contribute to improving the current state of knowledge regarding the enzyme inhibitory potential of the Bursera genus, but could also lead to the development of new sustainable bioinsecticides.

Circulating markers of oxidative stress are associated with a muscle injury in patients with muscular dystrophy Duchenne.

BACKGROUND: A small number of studies have confirmed the presence of oxidative damage in patients with Duchenne muscular dystrophy (DMD). Nevertheless, it is unknown if there a relationship of circulating markers of oxidative stress with a muscle injury. OBJECTIVE: We evaluated if oxidative damage and anti-oxidant markers are associated with muscle damage in DMD. METHODS: This cross-sectional study included 24 patients with DMD classified in ambulatory and non-ambulatory. Markers of muscle damage (creatine kinase [CK]), oxidative damage (malondialdehyde [MDA], and 8-isoprostane), anti-oxidant function (Thiol and mRNA of NRF2 and NF-κB) and nitric oxide (NO) were quantified in circulation. RESULTS: Total NO, MDA, and 8-isoprostane concentrations were significantly (p < 0.05) higher, and thiol concentration was lower in non-ambulatory than ambulatory patients. A significant correlation (p < 0.05) between muscle injury (evaluated by Vignos scale) with CK (r = -0.382), NO (r = 0.444), MDA (r = 0.503), 8-isoprostanes (r = 0.435) and thiol (r = -0.430) was observed. CONCLUSION: These findings suggest that non-ambulatory have high oxidative damage and low anti-oxidant function than ambulatory patients with DMD. Total nitric oxide and oxidative damage plasma markers increase, but the anti-oxidant marker thiol decreases with a muscle injury in boys with DMD. The findings of this study suggest that these markers could be considered as goods indicators of oxidative damage in longitudinal studies to evaluate the muscle injury during DMD progression. Additionally, these findings add new information about the pathophysiology of DMD.

Hepatic Glucose Output Inhibition by Mexican Plants Used in the Treatment of Type 2 Diabetes.

De novo hepatic glucose production or hepatic gluconeogenesis is the main contributor to hyperglycemia in the fasting state in patients with type 2 diabetes (T2D) owing to insulin resistance, which leads to at least twice as much glucose synthesis compared to healthy subjects. Therefore, control of this pathway is a promising target to avoid the chronic complications associated with elevated glucose levels. Patients with T2D in the rural communities of Mexico use medicinal plants prepared as infusions that are consumed over the day between meals, thus following this rationale (consumption of the infusions in the fasting state), one approach to understanding the possible mechanism of action of medicinal plants is to assess their capacity to inhibit hepatic glucose production. Furthermore, in several of these plants, the presence of phenolic acids able to block the enzyme glucose-6-phosphatase (G6Pase) is reported. In the present work, extracts of Ageratina petiolaris, Bromelia karatas, Equisetum myriochaetum, Rhizophora mangle, and Smilax moranensis, which are Mexican plants that have been traditionally used to treat T2D, were assayed to evaluate their possible hepatic glucose output (HGO) inhibitory activity with a pyruvate tolerance test in 18-h fasted STZ-NA Wistar rats after oral administration of the extracts. In addition, the in vitro effects of the extracts on the last HGO rate-limiting enzyme G6Pase was analyzed. Our results showed that four of these plants had an effect on hepatic glucose production in the in vivo or in vitro assays. A. petiolaris and R. mangle extracts decreased glucose output, preventing an increase in the blood glucose levels and sustaining this prevented increase after pyruvate administration. Moreover, both extracts inhibited the catalytic activity of the G6Pase complex. On the other hand, even though S. moranensis and B. karatas did not exhibit a significant in vivo effect, S. moranensis had the most potent inhibitory effect on this enzymatic system, while the E. myriochaetum extract only inhibited hepatic glucose production in the pyruvate tolerance test. Because of the traditional method in which diabetic patients use plants, hepatic glucose production inhibition seems to be a mechanism that partially explains the common hypoglycemic effect. However, further studies must be carried out to characterize other mechanisms whereby these plants can decrease HGO.

Hypoglycemic Activity of Medicinal Plants Used among the Cakchiquels in Guatemala for the Treatment of Type 2 Diabetes.

Type 2 diabetes (T2D) is a major health problem worldwide. In this condition, the organism can produce insulin but becomes resistant to it; thus the insulin is ineffective. High blood glucose levels are a result of insulin resistance and insulin deficiency; they produce diabetes-associated complications such as kidney failure, blindness, cardiovascular disease, and lower-limb amputation. In Guatemala, there were over 752.700 cases of the disease in 2017 with prevalence of 8.4 (IDF, 2017). The use of plants for medicinal purposes has been practiced in the country since pre-Hispanic times. Among the Cakchiquels, the aerial parts of Hamelia patens Jacq., Neurolaena lobata (L.) R.Br. ex Cass., and Solanum americanum Mill. and the cortex of Croton guatemalensis Lotsy are used to treat type 2 diabetes. The aim of the present study was to confirm the hypoglycemic effect of the plants under normal conditions and under maltose and sucrose tolerance tests, as well as to test the activity of the plant extracts in vitro against alpha-glucosidases types I and II. In agreement with the traditional usage of the plants, in normal conditions without a sugar load, the extracts produced a statistically significant hypoglycemic effect similar to the control drug glibenclamide. When the sugar load was maltose, only Croton and Solanum produced a statistically significant (p < 0.05) hypoglycemic effect compared to the control drug, but when the sugar was sucrose, Croton and Hamelia produced a statistically significant effect (p < 0.05) beginning at 30 min compared to the control group, while Solanum did so at 60 min and Neurolaena at 90 min. In vitro assays showed that the extracts inhibited yeast alpha-glucosidases but not the rat intestinal ones. Of the tested plants, Croton exert an effect both under sugars' tests and under a normal tolerance test; these results suggest the potential use of this plant. The results presented here provided evidence based on the use of these plants as hypoglycemic agents in the treatment of type 2 diabetes.

Creosote Bush (Larrea tridentata) Improves Insulin Sensitivity and Reduces Plasma and Hepatic Lipids in Hamsters Fed a High Fat and Cholesterol Diet.

Creosote bush, Larrea tridentata (Sesse y Moc. Ex DC, Zygophyllaceae) is a shrub found in the deserts of Northern Mexico and Southwestern United States. In traditional medicine, it is used to treat a variety of illnesses including type 2 diabetes. The present study aims to investigate the effects of creosote bush ethanolic extract on plasma and liver parameters associated with the metabolic syndrome in hamsters fed a high fat and cholesterol diet (HFD), comparing them with those induced by ezetimibe (EZ). Seven groups of six hamsters each were formed. Six groups were fed HFD for 2 weeks. The following 2 weeks, the HFD groups received: (1) only HFD, (2) HFD + 3 mg% EZ, (3) HFD + 0.2% creosote bush ethanolic extract, (4) only standard diet (Std Diet), (5) Std Diet + 3 mg% EZ, (6) Std Diet + 0.2% creosote bush ethanolic extract. The beneficial effects of creosote bush ethanolic extract in the HFD hamster model were a reduction of insulin resistance, associated with lower serum insulin and leptin, lower hepatic lipid peroxidation and higher liver antioxidant capacity. Plasma and liver lipids tended or were reduced to values closer to those of animals fed standard diet. A similar effect on lipids was induced by EZ, although with even lower hepatic cholesterol and total lipids concentrations. In general, the change from HFD to standard diet plus ethanolic extract induced the same but deeper changes, including a reduction in plasma glucose and an increase in the percentage of HDL cholesterol. Unlike creosote bush extract, EZ increased food consumption and neutral fecal steroids, with no significant effect on body weight, epididymal fat pads, liver peroxidation or antioxidant capacity. Also EZ did not modify serum insulin and leptin. However, insulin sensitivity improved to values similar to those induced by the extract. This suggests that the mechanism of action of creosote bush ethanolic extract is different to inhibition of cholesterol absorption or increase excretion. The ethanolic extract of L. tridentata could be useful in the treatment of the metabolic syndrome.

Evaluación de dieta purificada para la obtención de biomodelo de ratas anémicas / Evaluation of a purified diet to obtain a biomodel of anemic rats

Introducción: el desarrollo científico y tecnológico en las ciencias biológicas y biomédicas ha sido posible gracias a la utilización de modelos animales experimentales. La anemia por deficiencia de hierro (Fe), afecta aproximadamente a la tercera parte de la población mundial. La rata recién destetada ha sido el modelo animal más empleado en la obtención de un biomodelo experimental de anemia. Para la obtención de anemia se utiliza el método de depleción de la hemoglobina (Hb), mediante una alimentación basada en una dieta deficiente en Fe. Objetivo: el objetivo de este trabajo fue evaluar la eficacia de una dieta purificada a partir de caseína en la obtención de ratas anémicas. Métodos: se utilizaron 40 ratas Sprague Dawley recién destetadas (21-24 días), la mitad de cada sexo. Se alojaron en grupos de 3 del mismo sexo en cajas de acero inoxidable con piso de rejillas. Durante 8 semanas se les administró agua desionizada y una dieta de caseína al 12,58 por ciento de proteínas y 1,28 mgFe/Kg. Cada dos semanas se midió el peso corporal y a partir de la cuarta semana, la concentración de Hb. Resultados: se obtuvo un incremento significativo del peso corporal sin observarse diferencias entre hembras y machos. Se tomó como criterio de obtención de la anemia aquellos animales cuya Hb disminuyó al menos en un 30 por ciento del valor inicial promedio por grupo. El porcentaje de animales anémicos a las 8 semanas fue superior en las hembras (75 por ciento), mientras que en machos (60 por ciento). La dieta utilizada en este estudio permitió obtener ratas anémicas en el sexo hembras con una eficiencia aceptable, en relación al porcentaje de animales anémicos y al incremento del peso corporal. Conclusiones: la dieta utilizada en este estudio permitió obtener ratas anémicas en el sexo hembras con una eficiencia aceptable, en relación al porcentaje de animales anémicos y al incremento del peso corporal(AU)

Introduction: the scientific and technological development of biological and biomedical sciences has been possible due to the use of experimental animal models. Iron- (Fe-) deficiency anemia affects about one third of the world population. Just weaned rats have been the animal model most commonly used to obtain an experimental biomodel of anemia. The disease is obtained by the hemoglobin (Hb) depletion method, feeding the animal an iron-deficient diet. Objective: evaluate the efficacy of a purified diet based on casein to obtain anemic rats. Method: a study was conducted of 40 just weaned (21-24 days) Sprague Dawley rats, 20 of each sex. The animals were housed in groups of 3 rats of the same sex in stainless steel boxes with mesh bottoms. For 8 weeks the animals were administered deionized water and a casein diet of 12.58 percent protein and 1.28 mgFe/kg. Body weight was measured every two weeks. Hb concentration was gauged from the fourth week onwards. Results: a significant increase in body weight was obtained, with no differences between male and female rats. The criterion for anemia was at least a 30 percent decrease in Hb with respect to the average baseline value per group. The percentage of anemic animals at 8 weeks was higher for females (75 percent ) than for males (60 percent ) Conclusions: the diet used for the study made it possible to obtain female anemic rats with acceptable efficiency in relation to the percentage of anemic animals and the increase in body weight(AU)

Evaluación de dieta purificada para la obtención de biomodelo de ratas anémicas / Evaluation of a purified diet to obtain a biomodel of anemic rats

Introducción: el desarrollo científico y tecnológico en las ciencias biológicas y biomédicas ha sido posible gracias a la utilización de modelos animales experimentales. La anemia por deficiencia de hierro (Fe), afecta aproximadamente a la tercera parte de la población mundial. La rata recién destetada ha sido el modelo animal más empleado en la obtención de un biomodelo experimental de anemia. Para la obtención de anemia se utiliza el método de depleción de la hemoglobina (Hb), mediante una alimentación basada en una dieta deficiente en Fe. Objetivo: el objetivo de este trabajo fue evaluar la eficacia de una dieta purificada a partir de caseína en la obtención de ratas anémicas. Métodos: se utilizaron 40 ratas Sprague Dawley recién destetadas (21-24 días), la mitad de cada sexo. Se alojaron en grupos de 3 del mismo sexo en cajas de acero inoxidable con piso de rejillas. Durante 8 semanas se les administró agua desionizada y una dieta de caseína al 12,58 por ciento de proteínas y 1,28 mgFe/Kg. Cada dos semanas se midió el peso corporal y a partir de la cuarta semana, la concentración de Hb. Resultados: se obtuvo un incremento significativo del peso corporal sin observarse diferencias entre hembras y machos. Se tomó como criterio de obtención de la anemia aquellos animales cuya Hb disminuyó al menos en un 30 por ciento del valor inicial promedio por grupo. El porcentaje de animales anémicos a las 8 semanas fue superior en las hembras (75 por ciento), mientras que en machos (60 por ciento). La dieta utilizada en este estudio permitió obtener ratas anémicas en el sexo hembras con una eficiencia aceptable, en relación al porcentaje de animales anémicos y al incremento del peso corporal. Conclusiones: la dieta utilizada en este estudio permitió obtener ratas anémicas en el sexo hembras con una eficiencia aceptable, en relación al porcentaje de animales anémicos y al incremento del peso corporal

Introduction: the scientific and technological development of biological and biomedical sciences has been possible due to the use of experimental animal models. Iron- (Fe-) deficiency anemia affects about one third of the world population. Just weaned rats have been the animal model most commonly used to obtain an experimental biomodel of anemia. The disease is obtained by the hemoglobin (Hb) depletion method, feeding the animal an iron-deficient diet. Objective: evaluate the efficacy of a purified diet based on casein to obtain anemic rats. Method: a study was conducted of 40 just weaned (21-24 days) Sprague Dawley rats, 20 of each sex. The animals were housed in groups of 3 rats of the same sex in stainless steel boxes with mesh bottoms. For 8 weeks the animals were administered deionized water and a casein diet of 12.58 percent protein and 1.28 mgFe/kg. Body weight was measured every two weeks. Hb concentration was gauged from the fourth week onwards. Results: a significant increase in body weight was obtained, with no differences between male and female rats. The criterion for anemia was at least a 30 percent decrease in Hb with respect to the average baseline value per group. The percentage of anemic animals at 8 weeks was higher for females (75 percent ) than for males (60 percent ) Conclusions: the diet used for the study made it possible to obtain female anemic rats with acceptable efficiency in relation to the percentage of anemic animals and the increase in body weight

Alfa-glucosidase-inhibiting activity of some Mexican plants used in the treatment of type 2 diabetes.

Type 2 diabetes is an endocrine disease, which accounts for 9% of deaths worldwide. The aim of oral therapy is to reach normoglycemia to prevent later complications. Among glucose-lowering medications, alpha-glucosidase inhibitors delay the absorption of ingested carbohydrates, reducing the postprandial glucose and insulin peaks. In the present study, we tested the butanolic extracts of four Mexican plants with respect to their alpha-glucosidase inhibition activity, without excluding other possible mechanisms of action. The plants Cecropia obtusifolia Bertol., Equisetum myriochaetum Schlecht & Cham, Acosmium panamense (Benth.) Yacolev and Malmea depressa (Baill) R.E. Fries are used in traditional medicine to treat type 2 diabetes. In previous studies, we have demonstrated these plants' hypoglycemic activity and determined the phytochemical composition of their extracts. Our results in n-STZ diabetic rats loaded with maltose showed that Malmea and Acosmium extracts decreased plasma glucose significantly from 30 min on resembling the effect of acarbose. Cecropia extract produced the highest reduction of plasma glucose, and at 90 min, the glucose level was lower than the fasting level, which suggests another mechanism of action. Equisetum did not exert any effect. In vitro assays of alpha-glucosidase activity showed an IC(50) of 14 microg/ml for Cecropia, 21 microg/ml for Malmea, and 109 microg/ml for Acosmium, which were lower than that of acarbose (128 microg/ml). Equisetum did not show any significant effect on this assay, either. These results contribute to understand the mechanism of action of these plants on glucose metabolism.

The Harderian gland of the Mexican volcano mouse Neotomodon alstoni alstoni (Merriam 1898): a morphological and biochemical approach.

The Harderian glands of rodents are large intraorbital exocrine glands with histologic organization that varies among mammalian species. Here we describe some ultrastructural and biochemical features of the Harderian gland in the Mexican volcano mouse Neotomodon alstoni alstoni, a species of restricted habitat. The Harderian glands from male and female adult mice were dissected, processed and embedded in Epon 812 for light and electron microscopy studies. Porphyrin and total lipids were biochemically determined. The macroscopic appearance of the Harderian gland is similar in the male and female. The gland is a bilobulate structure, situated in the orbit towards the posterior side of the eyeball, of whitish color and is surrounded by a connective tissue capsule. The male gland is slightly heavier (127 mg) than that of the female (113 mg). The Harderian gland shows a tubulo-alveolar organization and is composed exclusively of one type of secretory cells. No branched duct system within the gland was found. Adrenergic nerves endings and mast cell were observed in the interstices of the alveoli. Male and female glands produce similar levels of porphyrins. Triglyceride levels were significantly higher (P < 0.05) in the female compared to the male. Abundance of lipids could induce corneal lubrication of the Harderian gland which may confer a protective and adaptative function to the volcano mouse in its natural habitat during the dry and cold seasons.

Pathophysiology of cholesterol gallstone disease

The purpose of the present paper is to review the current knowledge about cholesterol gallstone disease. It is generally accepted that the formation of cholesterol gallstone requires three major pathogenic defect, namely, supersaturation, nucleation and crystal growth as well as disorder of gallbladder motility. The supersaturation is necessary but not sufficient to explain stone formation. It has been suggested that nucleation is the key factor for gallstone formation. However, those three factors are necessary for the formation of cholesterol gallstones, and the presence of just one or two factor does not lead to stones. We also touch briefly on the results form studies performed in Mexico in this area