Effects of Full-Spectrum Cannabis Oil with a Cannabidiol:Tetrahydrocannabinol 2:1 Ratio on the Mechanisms Involved in Hepatic Steatosis and Oxidative Stress in Rats Fed a Sucrose-Rich Diet.

Introduction: This study aimed to analyze the effects of cannabis oil (cannabidiol:tetrahydrocannabinol [CBD:THC], 2:1 ratio) on the mechanisms involved in hepatic steatosis and oxidative stress in an experimental model of metabolic syndrome (MS) induced by a sucrose-rich diet (SRD). We hypothesized that noninvasive oral cannabis oil administration improves hepatic steatosis through a lower activity of lipogenic enzymes and an increase in carnitine palmitoyltransferase-1 (CPT-1) enzyme activity involved in the mitochondrial oxidation of fatty acids. Furthermore, cannabis oil ameliorates liver oxidative stress through the regulation of the main regulatory factors involved, nuclear factor erythroid 2 (NrF2) and nuclear factor-kB (NF-κB) p65. For testing this hypothesize, a relevant experimental model of MS was induced by feeding rats with a SRD for 3 weeks. Methods: Male Wistar rats were fed the following diets for 3 weeks: reference diet: standard commercial laboratory diet, SRD, and SRD + cannabis oil: noninvasive oral administration of 1 mg/kg body weight cannabis oil daily. The full-spectrum cannabis oil presents a total cannabinoid CBD:THC 2:1 ratio. Serum glucose, triglyceride, total cholesterol, HDL-cholesterol, LDL-cholesterol, uric acid, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase (AP), N-arachidonoylethanolamine or anandamide and 2-arachidonoylglycerol endocannabinoids levels, thiobarbituric acid reactive substance (TBARS) levels, and non-enzymatic antioxidant capacity (ferric ion-reducing antioxidant power [FRAP]) were evaluated. In the liver tissue: histology, nonalcoholic fatty liver disease activity score (NAS), triglycerides and cholesterol content, lipogenic enzyme activities (fatty acid synthase, acetyl-CoA carboxylase, malic enzyme, and glucose-6-phosphate dehydrogenase), enzyme related to mitochondrial fatty acid oxidation (CPT-1), reactive oxygen species, TBARS, FRAP, glutathione, catalase, glutathione peroxidase, and glutathione reductase enzyme activities. 4-hydroxynonenal, NrF2, and NF-κB p65 levels were analyzed by immunohistochemistry. Results: The results showed that SRD-fed rats developed dyslipidemia, liver damage, hepatic steatosis (increase of key enzymes related to the novo fatty acid synthesis and decrease of key enzyme related to mitochondrial fatty acid oxidation), lipid peroxidation, and oxidative stress. Hepatic NrF2 expression was significantly decreased and NF-κB p65 expression was increased. Cannabis oil administration improved dyslipidemia, liver damage, hepatic steatosis, lipid peroxidation (improving enzymes involved in lipid metabolism), and oxidative stress. In the liver tissue, NrF2 expression increased, and NF-κB p65 expression was reduced. Conclusion: The present study revealed new aspects of liver damage and steatosis, lipid peroxidation, and oxidative stress in dyslipidemic insulin-resistant SRD-fed rats. We demonstrated new properties and molecular mechanisms of cannabis oil (CBD:THC, 2:1 ratio) on lipotoxicity and hepatic oxidative stress in an experimental model of MS.

Salvia hispanica L. (chia) seed improves liver inflammation and endothelial dysfunction in an experimental model of metabolic syndrome.

The purposes of the present study were to analyze liver inflammation and endothelial dysfunction in an experimental model of metabolic syndrome (MS) induced by chronic administration of a sucrose-rich diet (SRD) and to evaluate the effects of chia seed as a therapeutic strategy. Male Wistar rats were fed with a reference diet (RD) for 6 months or a SRD for 3 months. Then, the latter group was randomly divided into two subgroups. One subgroup continued receiving the SRD for up to 6 months and the other was fed with a SRD where whole chia seed was incorporated as a source of dietary fat for the next 3 months (SRD + CHIA). Results showed that rats fed a SRD for a long period of time developed dyslipidemia, hyperglycemia, inflammation and endothelial dysfunction. Hepatic NAS, IL-1ß, NFκB p65, PAI-1, and F4-80 expression, as well as MPO activity were significantly increased and IL-10 expression was significantly decreased; this was accompanied by increased plasma IL-6 and TNF-α levels in rats fed a SRD. In addition, serum and liver nitric oxide (NO) levels and nitric oxide synthase (NOS) were significantly increased in the SRD group. In addition, a significant increase in hepatic iNOS expression and a positive correlation of this with liver NFκB p65 was found. We observed a significant increase in hepatic intercellular adhesion molecule (ICAM), and a negative correlation of this with liver Nrf2 was found. The administration of chia seed for 3 months reversed dyslipidemia, hyperglycemia, inflammation and endothelial dysfunction. In the liver tissue, NAS, IL-1ß, IL-10, NFκB p65, PAI-1, and F4-80 expression and MPO activity were normalized. Serum and liver NO and NOS levels and hepatic iNOS expression were decreased and this last one was associated with a decrease in liver NFκB p65 levels. Hepatic ICAM-1 was normalized and negatively correlated with liver NrF2 levels. This study showed new aspects of liver inflammation and endothelial dysfunction in dyslipidemic insulin resistant rats chronically fed with a sucrose-rich diet. In addition, we demonstrated new properties and molecular mechanisms associated with beneficial effects on inflammation and endothelial dysfunction of chia seed as a therapeutic strategy.

Salvia hispanica L. (chia) seed ameliorates liver injury and oxidative stress by modulating NrF2 and NFκB expression in sucrose-rich diet-fed rats.

The aim of this study was to analyze the liver injury and oxidative stress in an experimental model of Metabolic Syndrome (MS) induced by chronic administration of a sucrose-rich diet (SRD) and to evaluate the effects of chia seed as a therapeutic strategy. Male Wistar rats were fed with a reference diet (RD) -6 months- or a SRD -3 months. Then, the latter group was randomly divided into two subgroups. One subgroup continued receiving the SRD for up to 6 months and the other was fed with a SRD where whole chia seed was incorporated as a source of dietary fat for the next 3 months (SRD+CHIA). The results showed that rats fed with a SRD for a long period of time developed dyslipidemia, hyperglycemia, hepatic lipid accumulation, liver injury, hepatic lipid peroxidation and oxidative stress. Hepatic NrF2 expression was significantly decreased. In addition, a significant increase in hepatic NFκB p65 expression and a positive correlation of this with plasma TNFα levels were found. The administration of chia seed for 3 months reversed dyslipidemia, hyperglycemia, lipid accumulation, liver injury, lipid peroxidation and oxidative stress. In the liver tissue, NrF2 expression was normalized and NFκB p65 expression was decreased, the latter was associated with a decrease in plasma TNFα levels. The present study showed new aspects of liver damage, lipid peroxidation and oxidative stress in dyslipidemic insulin resistant rats chronically fed with a sucrose-rich diet. However, we demonstrated new properties and molecular mechanisms associated with the beneficial anti-oxidant effects of chia seed consumption.

Effects of Salvia hispanica L. (chia) seed on blood coagulation, endothelial dysfunction and liver fibrosis in an experimental model of Metabolic Syndrome.

The aim of this study was to analyze blood coagulation, endothelial dysfunction and liver fibrosis in an experimental model of Metabolic Syndrome (MS) induced by chronic administration of a sucrose-rich diet (SRD) and to evaluate the effects of chia seed as a therapeutic strategy. Male Wistar rats were fed with a reference diet (RD) - 6 months - or a SRD - 3 months. Then, the last group was randomly divided into two subgroups. One subgroup continued receiving the SRD for up to 6 months and the other was fed with a SRD where whole chia seed was incorporated as the source of dietary fat for the next 3 months (SRD + CHIA). Results showed that rats fed a SRD for a long period of time develop dyslipidemia, visceral adiposity, insulin resistance, and a hypercoagulable and hypofibrinolytic basal state. Hepatic VCAM-1 (main adhesion molecules involved in endothelial dysfunction) expression was significantly increased. In addition, the SRD group presented hepatic steatosis, a significant increase in interstitial collagen deposition and hydroxyproline content. Liver TGF-ß1 (a key cytokine involved in fibrogenesis) levels increased and a negative correlation with PPARα protein mass levels was found. The administration of chia seed for 3 months reversed dyslipidemia, visceral adiposity and insulin resistance. Platelet count, coagulation parameters and plasma fibrinogen levels were normalized. In the liver tissue, VCAM-1 expression, steatosis, interstitial collagen deposition and the hydroxyproline content decreased. TGF-ß1 expression was decreased and this was associated with an increase in the PPARα protein levels. The present study showed new aspects in the progression from liver steatosis to fibrosis in dyslipidemic insulin-resistant rats chronically fed a sucrose-rich diet. Chia seed supplementation could be used as a functional food and a potential dietary strategy to prevent or ameliorate disorders related to atherothrombotic cardiovascular events and NASH.

Salvia hispanica L. (chia) seed promotes body fat depletion and modulates adipocyte lipid handling in sucrose-rich diet-fed rats.

The aim of this study was to analyze the effects of Salvia hispanica L. (chia) seed upon metabolic pathways that play a key role in adipose tissue lipid handling which could be involved in visceral adiposity reduction developed in rats fed a sucrose-rich diet (SRD). Male Wistar rats were fed with a reference diet (RD) -6 months- or SRD-3 months. Then, the last group was randomly divided into two subgroups. One subgroup continued receiving the SRD up to 6 months and the other was fed with a SRD where whole chia seed was incorporated as the source of dietary fat for the next 3 months (SRD + CHIA). Results showed that chia seed in the SRD-fed rat reduced the abdominal and thoracic circumferences, carcass fat content, adipose tissue weights, and visceral adiposity index. This was accompanied by an improvement in insulin sensitivity and plasma lipid profile. In epididymal adipose tissue, the decreased fat cell triglyceride content was associated with a reduction in both, FAT/CD 36 plasma membrane levels and the fat synthesis enzyme activities. There were not changes in oxidative CPT enzyme activities. PKCß and the precursor and mature forms of SREBP-1 protein levels were decreased, while pAMPK was increased. Our findings suggest that chia seed supplementation can modulate essential pathways of lipid metabolism in adipose tissue, contributing to reduced visceral fat accumulation in SRD-fed rats.

Ingesta de calcio en jóvenes estudiantes de Licenciatura en Nutrición. Santa Fe (Argentina) / Calcium intake in young students of Nutrition. Santa Fe (Argentina)

INTRODUCCIÓN: En Argentina, el 94% de la población nocumple las recomendaciones para el consumo de alimentos fuente de calcio (Ca), cuya ingesta se correlaciona con la salud ósea e inversamente con el índice de masa corporal y desarrollo de sobrepeso y obesidad. OBJETIVO: Evaluar la ingesta de Ca, estado nutricional, realización de actividad física (AF) y antecedentes familiares, en mujeres estudiantes de la carrera de Licenciatura en Nutrición. MÉTODOS: Estudio descriptivo-transversal en 123 universitarias. Se midieron peso, talla y circunferencia de cintura (CC). Se indagó sobre antecedentes familiares y estilo de vida. Mediante 3 recordatorios de 24 horas se evaluó la distribución diaria de comidas, tipo de lácteos incorporados en el desayuno e ingesta habitual de Ca total y Ca lácteo. Se utilizó el Sistema de Análisis y Registro de los Alimentos del Ministerio de Salud de la Nación y los resultados se presentaron con frecuencias absolutas y relativas, utilizando medidas de tendencia central y de dispersión. RESULTADOS: El 8,1% tuvo antecedentes de 1° grado de enfermedad ósea, y el 19,5%, de 2° grado. El 17,1% presentó exceso de peso. La CC mostró un 11,4% de riesgo cardiovascular "aumentado o muy aumentado". El 35% no cumplió con las recomendaciones de AF. Casi la totalidad de las estudiantes manifestaron el hábito de desayunar, sin embargo, un 21,1% no incorporó lácteos en dicha comida. La ingesta de Ca total fue de 693,79±208,95 mg/día, y 438,23±194,53 mg/día proveniente de alimentos lácteos, representando el 60,93±13,50%. Menos del 10% de las estudiantes cumplieron con las recomendaciones de 1.000 mgCa/día. CONCLUSIÓN: Casi la totalidad de la población estudiada, a pesar de ser futuras profesionales de la nutrición, no cumple con las recomendaciones de consumo de Ca establecidas por las Guías Alimentarias para la Población Argentina

INTRODUCTION: In Argentina, 94% of the population does not meet the recommendations for the consumption of food sources of calcium (Ca), whose intake correlates with bone health and inversely with the body mass index and development of overweight and obesity. OBJECTIVE: To evaluate Ca intake, nutritional status, physical activity (PA) performance, and family history, in female students of Nutrition degree. METHODS: Descriptive-cross-sectional study in 123 university students. Weight, height, and waist circumference (CC) were measured. Family history and lifestyle were investigated. Through 3 reminders of 24 hours, the daily distribution of meals, type of dairy incorporated in breakfast and habitual intake of total Ca and dairy Ca were evaluated. The Food Analysis and Registration System of the National Ministry of Health was used, and the results were presented with absolute and relative frequencies, using measures of central tendency and dispersion. RESULTS: 8.1% had a history of 1st degree of bone disease, and 19.5% of 2nd degree. 17.1% presented excess weight. The CC showed an 11.4% cardiovascular risk "increased or greatly increased". 35% did not comply with the recommendations for PA. Almost all of the students manifested the habit of having breakfast, however, 21.1% did not incorporate dairy in said food. Total Ca intake was 693.79 ± 208.95 mg/day, and 438.23 ± 194.53 mg/day from dairy foods, representing 60.93 ± 13.50%. Less than 10% of the students met the recommendations of 1,000 mg Ca/day. CONCLUSION: Almost the entire population studied, despite being future nutrition professionals, does not comply with the recommendations for Ca consumption established by the Dietary Guidelines for the Argentine Population

Generation and characterization of adeno-associated virus producer cell lines for research and preclinical vector production.

Adeno-associated virus (AAV) producer cell lines represent an effective method for large-scale production of AAV vectors. We set out to evaluate and characterize the use of an abbreviated protocol to generate "masterwells" (MWs; a nonclonal cell population) as a platform for research and preclinical vector production. In this system, a single plasmid containing three components, the vector sequence, the AAV rep, and cap genes, and a selectable marker gene is stably transfected into HeLaS3 cells. Producer cell lines generating an AAV2 vector expressing a secreted form of human placental alkaline phosphatase (SEAP) have been created. Several MWs showed vector yields in the 5×10(4) to 2×10(5) DNase-resistant particles/cell range, and the productivity was stable over >60 population doublings. Integrated plasmid copy number in three high-producing MWs ranged from approximately 12 to 50; copies were arranged in a head-to-tail configuration. Upon infection with adenovirus, rep/cap copy number was amplified approximately 100-fold and high yield appeared to be dependent on the extent of amplification. Rep/cap gene expression and vector packaging both reached a peak at 48 hr postinfection. AAV2-SEAP vector was produced in 1-liter shaker culture and purified for assessment of vector quality and potency. The data showed that the majority of the capsids from the MWs contained vector DNA (≥70%) and that purified vector was free of replication-competent AAV. In vitro and in vivo analyses demonstrated that potency of the producer cell-derived vector was comparable to vector generated via the standard transfection method.

A finite element model for describing the effect of muscle shortening on surface EMG.

A finite-element model for the generation of single fiber action potentials in a muscle undergoing various degrees of fiber shortening is developed. The muscle is assumed fusiform with muscle fibers following a curvilinear path described by a Gaussian function. Different degrees of fiber shortening are simulated by changing the parameters of the fiber path and maintaining the volume of the muscle constant. The conductivity tensor is adapted to the muscle fiber orientation. In each point of the volume conductor, the conductivity of the muscle tissue in the direction of the fiber is larger than that in the transversal direction. Thus, the conductivity tensor changes point-by-point with fiber shortening, adapting to the fiber paths. An analytical derivation of the conductivity tensor is provided. The volume conductor is then studied with a finite-element approach using the analytically derived conductivity tensor. Representative simulations of single fiber action potentials with the muscle at different degrees of shortening are presented. It is shown that the geometrical changes in the muscle, which imply changes in the conductivity tensor, determine important variations in action potential shape, thus affecting its amplitude and frequency content. The model provides a new tool for interpreting surface EMG signal features with changes in muscle geometry, as it happens during dynamic contractions.