Biomarkers in Systemic Lupus Erythematosus along with Metabolic Syndrome.

Metabolic syndrome (MetS) is a group of physiological abnormalities characterized by obesity, insulin resistance (IR), and hypertriglyceridemia, which carry the risk of developing cardiovascular disease (CVD) and type 2 diabetes (T2D). Immune and metabolic alterations have been observed in MetS and are associated with autoimmune development. Systemic lupus erythematosus (SLE) is an autoimmune disease caused by a complex interaction of environmental, hormonal, and genetic factors and hyperactivation of immune cells. Patients with SLE have a high prevalence of MetS, in which elevated CVD is observed. Among the efforts of multidisciplinary healthcare teams to make an early diagnosis, a wide variety of factors have been considered and associated with the generation of biomarkers. This review aimed to elucidate some primary biomarkers and propose a set of assessments to improve the projection of the diagnosis and evolution of patients. These biomarkers include metabolic profiles, cytokines, cardiovascular tests, and microRNAs (miRs), which have been observed to be dysregulated in these patients and associated with outcomes.

Osteopontin: A Bone-Derived Protein Involved in Rheumatoid Arthritis and Osteoarthritis Immunopathology.

Osteopontin (OPN) is a bone-derived phosphoglycoprotein related to physiological and pathological mechanisms that nowadays has gained relevance due to its role in the immune system response to chronic degenerative diseases, including rheumatoid arthritis (RA) and osteoarthritis (OA). OPN is an extracellular matrix (ECM) glycoprotein that plays a critical role in bone remodeling. Therefore, it is an effector molecule that promotes joint and cartilage destruction observed in clinical studies, in vitro assays, and animal models of RA and OA. Since OPN undergoes multiple modifications, including posttranslational changes, proteolytic cleavage, and binding to a wide range of receptors, the mechanisms by which it produces its effects, in some cases, remain unclear. Although there is strong evidence that OPN contributes significantly to the immunopathology of RA and OA when considering it as a common denominator molecule, some experimental trial results argue for its protective role in rheumatic diseases. Elucidating in detail OPN involvement in bone and cartilage degeneration is of interest to the field of rheumatology. This review aims to provide evidence of the OPN's multifaceted role in promoting joint and cartilage destruction and propose it as a common denominator of AR and OA immunopathology.

Osteopontin: another piece in the systemic lupus erythematosus immunopathology puzzle.

Osteopontin (OPN) is a phosphoglycoprotein involved in bone remodelling, wound healing, cell adhesion, tissue remodelling, and immune response that is distributed widely in normal adult tissues. OPN biological activity is regulated by thrombin and matrix metalloproteinases (MMPs) cleavage, where the full-length (OPN-FL) protein and the cleaved OPN-N are associated with autoimmune diseases such as systemic lupus erythematosus (SLE). OPN overexpression has been associated with a predisposition to SLE and bad prognosis since OPN could mediate a sustained polyclonal B cell activation that besides to intracellular OPN (iOPN) form, promote the T follicular helper (TFH) cells and enhance anti-nuclear antibody production. Currently, the role of OPN in lupus nephritis (LN) has been reported and extensively studied; however, no data are available about the potential mechanism of OPN in neuropsychiatric SLE (NPSLE). In this review, we highlighted the contribution of OPN and iOPN in LN and NPSLE immunopathology.

The DNA co-vaccination using Sm antigen and IL-10 as prophylactic experimental therapy ameliorates nephritis in a model of lupus induced by pristane.

INTRODUCTION: Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the production of autoantibodies such as anti-Sm. Studies in patients with SLE and murine models of lupus reveal that the most critical anti-Sm autoantibodies are predominantly direct against D1(83-119), D2, and B´/B epitopes. OBJECTIVES: The present study aimed to analyze the induction of antigen-specific tolerance after prophylactic immunization with a DNA vaccine encoding the epitopes: D183-119, D2, B´/B, and B´/BCOOH in co-vaccination with IFN-γ or IL-10 in a murine model of lupus induced by pristane. MATERIAL AND METHODS: To obtain endotoxin-free DNA vaccines, direct cloning techniques using pcDNA were performed: D183-119, D2, B´/B, B´/BCOOH, IFN-γ, or IL-10. Lupus was induced by 0.5 mL of pristane via intraperitoneal in BALB/c female mice. Immunoprecipitation with K562 cells was metabolically labeled with 35S and ELISA to detect serum antibodies or mice IgG1, IgG2a isotypes. ELISA determined IL-10 and IFN-γ from splenocytes supernatants. Proteinuria was assessed monthly, and lupus nephritis was evaluated by immunofluorescence, and electron microscopy. RESULTS: The prophylactic co-vaccination with D2/IL-10 reduced the expression of kidney damage observed by electron microscopy, direct immunofluorescence, and H & E, along with reduced level of anti-nRNP/Sm antibodies (P = 0.048). CONCLUSION: The prophylactic co-vaccination of IL-10 with D2 in pristane-induced lupus ameliorates the renal damage maybe by acting as prophylactic DNA tolerizing therapy.

Contribution of rs3211938 polymorphism at CD36 to glucose levels, oxidized low-density lipoproteins, insulin resistance, and body mass index in Mexican mestizos with type-2 diabetes from western Mexico / Contribución del polimorfismo rs3211938 de CD36 a los niveles de glucosa, las lipoproteínas de baja densidad oxidadas, la resistencia a la insulina y el índice de masa corporal en mestizos mexicanos con diabetes de tipo 2 del occidente de México

Background: type-2 diabetes mellitus (T2DM) is a chronic metabolic disorder characterized by hyperglycemia, insulin resistance (IR), and abnormal fatty acid metabolism in which the CD36 receptor has been implicated in glucose and lipid dysregulation. Objective: to evaluate the contribution of polymorphism CD36 rs3211938 to metabolic profile in T2DM Mexican mestizos from western Mexico. Methods: we included 115 individuals classified as non-T2DM (NT2DM) adults and T2DM patients. Polymorphism CD36 rs3211938 was assessed by PCR-RFLP. Anthropometric and metabolic markers were measured by routine methods, and insulin and oxidized LDL (ox-LDL) were measured by ELISA. Results: the distribution of genotypes between NT2DM and T2DM patients was different (p < 0.001), as was the allele frequency (p = 0.002). NT2DM TG carriers showed the lowest levels of basal insulin and HOMA-IR index in comparison with TT carriers (p < 0.05 and p < 0.05, respectively). In the T2DM group TG carriers showed high BMI, WHR, and weight values (p = 0.001; p ≤ 0.05 and p < 0.05, respectively), and the highest levels of basal glucose, HDL-cholesterol, ox-LDL, and HOMA-IR (p < 0.001; p < 0.001; p < 0.001, and p = 0.001, respectively) in comparison with diabetic TT carriers. Conclusion: the CD36 rs3211938 TG genotype is associated with high levels of glucose, ox-LDL, HDL-cholesterol, and IR, and with increased BMI in Mexican mestizo T2DM patients from western Mexico. (AU)

Antecedentes: la diabetes mellitus de tipo 2 (DMT2) es un trastorno metabólico crónico caracterizado por hiperglucemia, resistencia a la insulina (RI) y metabolismo anormal de ácidos grasos en el que se ha implicado el receptor CD36 en la disregulación de la glucosa y los lípidos. Objetivo: evaluar la contribución del polimorfismo CD36 rs3211938 al perfil metabólico en individuos mestizos mexicanos con DMT2 del occidente de México. Métodos: se incluyeron 115 individuos clasificados en adultos sin DMT2 (NDMT2) y pacientes con DMT2. El polimorfismo CD36 rs3211938 se identificó mediante PCR-RFLP. Las mediciones antropométricas y metabólicas se realizaron mediante métodos de rutina y la insulina y las LDL-oxidadas (LDL-ox) se midieron por ELISA. Resultados: las distribuciones de los genotipos entre los pacientes NDMT2 y DMT2 fueron diferentes (p < 0,001), así como la frecuencia alélica (p = 0,002). Los individuos NDMT2 portadores del genotipo TG mostraron niveles más bajos de insulina basal e índice HOMA-IR en comparación con los portadores del genotipo TT (p < 0,05 y p < 0,05, respectivamente). En el grupo DMT2, los portadores del genotipo TG presentaron valores elevados de índice de masa corporal (IMC), índice cintura-cadera (ICC) y peso (p = 0,001; p < 0,05 y p < 0,05, respectivamente) y niveles más altos de glucosa basal, HDL-colesterol, LDL-ox y HOMA-IR (p < 0,001; p < 0,001; p < 0,001 y p = 0,001, respectivamente) en comparación con los portadores del genotipo TT. Conclusión: el genotipo TG del polimorfismo CD36 rs3211938 se asocia a altos niveles de glucosa, ox-LDL, HDL-colesterol y RI, y a aumentos del IMC en los pacientes mestizos mexicanos con DMT2 del occidente de México. (AU)

Contribution of rs3211938 polymorphism at CD36 to glucose levels, oxidized low-density lipoproteins, insulin resistance, and body mass index in Mexican mestizos with type-2 diabetes from western Mexico.

INTRODUCTION: Background: type-2 diabetes mellitus (T2DM) is a chronic metabolic disorder characterized by hyperglycemia, insulin resistance (IR), and abnormal fatty acid metabolism in which the CD36 receptor has been implicated in glucose and lipid dysregulation. Objective: to evaluate the contribution of polymorphism CD36 rs3211938 to metabolic profile in T2DM Mexican mestizos from western Mexico. Methods: we included 115 individuals classified as non-T2DM (NT2DM) adults and T2DM patients. Polymorphism CD36 rs3211938 was assessed by PCR-RFLP. Anthropometric and metabolic markers were measured by routine methods, and insulin and oxidized LDL (ox-LDL) were measured by ELISA. Results: the distribution of genotypes between NT2DM and T2DM patients was different (p < 0.001), as was the allele frequency (p = 0.002). NT2DM TG carriers showed the lowest levels of basal insulin and HOMA-IR index in comparison with TT carriers (p < 0.05 and p < 0.05, respectively). In the T2DM group TG carriers showed high BMI, WHR, and weight values (p = 0.001; p ≤ 0.05 and p < 0.05, respectively), and the highest levels of basal glucose, HDL-cholesterol, ox-LDL, and HOMA-IR (p < 0.001; p < 0.001; p < 0.001, and p = 0.001, respectively) in comparison with diabetic TT carriers. Conclusion: the CD36 rs3211938 TG genotype is associated with high levels of glucose, ox-LDL, HDL-cholesterol, and IR, and with increased BMI in Mexican mestizo T2DM patients from western Mexico.

INTRODUCCIÓN: Antecedentes: la diabetes mellitus de tipo 2 (DMT2) es un trastorno metabólico crónico caracterizado por hiperglucemia, resistencia a la insulina (RI) y metabolismo anormal de ácidos grasos en el que se ha implicado el receptor CD36 en la disregulación de la glucosa y los lípidos. Objetivo: evaluar la contribución del polimorfismo CD36 rs3211938 al perfil metabólico en individuos mestizos mexicanos con DMT2 del occidente de México. Métodos: se incluyeron 115 individuos clasificados en adultos sin DMT2 (NDMT2) y pacientes con DMT2. El polimorfismo CD36 rs3211938 se identificó mediante PCR-RFLP. Las mediciones antropométricas y metabólicas se realizaron mediante métodos de rutina y la insulina y las LDL-oxidadas (LDL-ox) se midieron por ELISA. Resultados: las distribuciones de los genotipos entre los pacientes NDMT2 y DMT2 fueron diferentes (p < 0,001), así como la frecuencia alélica (p = 0,002). Los individuos NDMT2 portadores del genotipo TG mostraron niveles más bajos de insulina basal e índice HOMA-IR en comparación con los portadores del genotipo TT (p < 0,05 y p < 0,05, respectivamente). En el grupo DMT2, los portadores del genotipo TG presentaron valores elevados de índice de masa corporal (IMC), índice cintura-cadera (ICC) y peso (p = 0,001; p < 0,05 y p < 0,05, respectivamente) y niveles más altos de glucosa basal, HDL-colesterol, LDL-ox y HOMA-IR (p < 0,001; p < 0,001; p < 0,001 y p = 0,001, respectivamente) en comparación con los portadores del genotipo TT. Conclusión: el genotipo TG del polimorfismo CD36 rs3211938 se asocia a altos niveles de glucosa, ox-LDL, HDL-colesterol y RI, y a aumentos del IMC en los pacientes mestizos mexicanos con DMT2 del occidente de México.

Presence of anti-TIF-1γ, anti-Ro52, anti-SSA/Ro60 and anti-Su/Ago2 antibodies in breast cancer: a cross-sectional study.

OBJECTIVES: The presence of myositis-specific antibodies (MSA), was recently reported in healthy individuals, cancer patients without myopathy and paraneoplastic rheumatic syndromes. We sought to analyze the frequency of MSA, myositis-associated antibodies (MAA) and autoantibodies related to systemic autoimmune rheumatic diseases (SARD) in breast cancer patients. METHODS: One hundred fifty-two breast cancer patients were enrolled in a cross-sectional study. Clinical information was collected, and autoantibodies tested by immunoprecipitation of an 35S-methionine-labeled K562 cell extract, enzyme-linked immunosorbent assay (ELISA) and Western blot when indicated. All statistical tests were performed using the software statistical package for the social science (SPSS) ver. 19.0 (IBM Inc., NYSE, USA). RESULTS: Autoantibodies associated with SARD: anti-52 kD ribonucleoprotein/tripartite motif-containing 21 (anti-Ro52/TRIM21) was found in 5.9% (9/152), anti-Sjögren syndrome-related antigen A/60 kD ribonucleoprotein antibody (anti-SSA/Ro60) in 3.9% (6/152) and anti-Su antigen/Argonaute 2 antibody (anti-Su/Ago2) in 2.6% (4/152). Meanwhile, anti-transcription intermediary factor-1γ (anti-TIF-1γ, p155/140) antibody was positive in 2 cases and anti-polymyositis/scleroderma antibody was detected in one case. As a whole, 14.47% (22/152) of breast cancer patients showed autoantibodies associated with SARD. These specific autoantibodies were not associated with the presence of rheumatic diseases except one rheumatoid arthritis patient positive for anti-Ro52/TRIM21. CONCLUSIONS: Autoantibodies to TIF-1γ were found in two patients with breast cancer without dermatomyositis (DM). More common specificities were autoantibodies anti-SSA/Ro60, anti-Ro52/TRIM21 and anti-Su/Ago2. More studies are needed in order to establish the biological meaning of the presence of SARD-associated autoantibodies in breast cancer.

In vivo assessment and characterization of lactic acid bacteria with probiotic profile isolated from human milk powder / Evaluación y caracterización in vivo de bacterias acidolácticas con perfil probiótico aisladas a partir de leche materna en polvo

INTRODUCTION: breast milk (MH) contains nutrients and bioactive compounds for child development, including probiotic bacteria, which contribute to intestinal maturation. This benefit accompanies the individual until adulthood. There are new methods such as spray drying that give this compound a good conservation without loss of microbiota. OBJECTIVE: the aim of this study was to analyze the viability of lactic acid bacteria isolated from human milk with probiotic potential after the spray drying process, as well as to evaluate the possible adhesion in the colon of mice of the Balb/C strain after feeding them powdered human milk and a commercial formula milk. METHOD: we isolated and identified the presence of lactic acid bacteria with possible probiotic potential in powdered human milk using the MALDI-TOF MS technique. Powdered human milk and a commercial formula milk were fed to mice of the Bald/C strain for 14 weeks. Glucose level and weight were measured in the mice. The feces were collected to verify the presence of lactic bacteria. The mice were sacrificed and their intestines were weighed, isolating the lactic acid bacteria both from the intestines and from the feces. The strains isolated from mice fed human milk were evaluated for their probiotic potential, analyzing their ability to inhibit pathogens, resistance to pH, temperature, adhesion, and hydrophobicity. RESULTS: the presence of Lactobacillus fermentum LH01, Lactobacillus rhamnosus LH02, Lactobacullis reuteri LH03, and Lactobacillus plantarum LH05 in powdered human milk was identified. All strains showed a possible probiotic profile due to the ability of bacteria to resist low pH, bile salts, and exposure to gastric enzymes, as well as their hydrophobicity and self-aggregation capacity, and their failure to show hemagglutination or hemolysis activity in a culture medium rich in erythrocytes. We observed that the consumption of powdered human milk prevented weight gain and constipation in mice. CONCLUSIONS: after spray drying, strains with possible probiotic potential may be preserved in human milk. The consumption of powdered human milk with probiotic bacteria prevents constipation and weight gain in mice, when compared to those fed a commercial formula milk

INTRODUCCIÓN: la leche materna (HM) contiene los nutrientes y compuestos bioactivos necesarios para el desarrollo infantil, incluidas bacterias probióticas, que contribuyen a la maduración intestinal. OBJETIVO: el objetivo de este estudio fue analizar la viabilidad de las bacterias acidolácticas aisladas de la leche humana con potencial probiótico, después del proceso de secado, así como evaluar su posible adhesión en el colón de ratones (BAlb/C) alimentados con leche humana en polvo y leche de una fórmula comercial. MÉTODO: se aislaron e identificaron mediante la técnica de Maldi-Tof-MS las bacterias acidolácticas con posible potencial probiótico en la leche humana en polvo. Se alimentó con leche humana en polvo y leche de una fórmula comercial a ratones de la cepa Bald/C durante 14 semanas. Se midieron el nivel de glucosa y el peso. Las heces se recolectaron para verificar la presencia de bacterias lácticas. Los ratones se sacrificaron y se pesaron los intestinos, aislando las bacterias lácticas tanto de los intestinos como de las heces. En las cepas aisladas de la leche humana se evaluó el potencial probiótico analizando su capacidad para inhibir patógenos, resistir distintos pH y temperaturas, adherirse y mostrar hidrofobicidad. RESULTADOS: se identificó la presencia de Lactobacillus fermentum LH01, Lactobacillus rhamnosus LH02, Lactobacullis reuteri LH03 y L. plantarum LH05 en la leche humana en polvo. Todas las cepas mostraron resistencia a los pH bajos, a las sales biliares y a la exposición a enzimas gástricas, así como una buena hidrofobicidad y capacidad de autoagregación. Además, no presentaron actividad de hemaglutinación o hemólisis en un medio de cultivo rico en eritrocitos. Observamos que el consumo de leche humana en polvo evita en los ratones el aumento de peso y el estreñimiento. CONCLUSIONES: después del secado por aspersión, las cepas con posible potencial probiótico pueden conservarse en la leche materna. El consumo de leche humana en polvo con bacterias probióticas evita el estreñimiento y el aumento de peso en los ratones, en comparación con los alimentados con leche de una formula comercial

In vivo assessment and characterization of lactic acid bacteria with probiotic profile isolated from human milk powder.

INTRODUCTION: Introduction: breast milk (MH) contains nutrients and bioactive compounds for child development, including probiotic bacteria, which contribute to intestinal maturation. This benefit accompanies the individual until adulthood. There are new methods such as spray drying that give this compound a good conservation without loss of microbiota. Objective: the aim of this study was to analyze the viability of lactic acid bacteria isolated from human milk with probiotic potential after the spray drying process, as well as to evaluate the possible adhesion in the colon of mice of the Balb/C strain after feeding them powdered human milk and a commercial formula milk. Method: we isolated and identified the presence of lactic acid bacteria with possible probiotic potential in powdered human milk using the MALDI-TOF MS technique. Powdered human milk and a commercial formula milk were fed to mice of the Bald/C strain for 14 weeks. Glucose level and weight were measured in the mice. The feces were collected to verify the presence of lactic bacteria. The mice were sacrificed and their intestines were weighed, isolating the lactic acid bacteria both from the intestines and from the feces. The strains isolated from mice fed human milk were evaluated for their probiotic potential, analyzing their ability to inhibit pathogens, resistance to pH, temperature, adhesion, and hydrophobicity. Results: the presence of Lactobacillus fermentum LH01, Lactobacillus rhamnosus LH02, Lactobacullis reuteri LH03, and Lactobacillus plantarum LH05 in powdered human milk was identified. All strains showed a possible probiotic profile due to the ability of bacteria to resist low pH, bile salts, and exposure to gastric enzymes, as well as their hydrophobicity and self-aggregation capacity, and their failure to show hemagglutination or hemolysis activity in a culture medium rich in erythrocytes. We observed that the consumption of powdered human milk prevented weight gain and constipation in mice. Conclusions: after spray drying, strains with possible probiotic potential may be preserved in human milk. The consumption of powdered human milk with probiotic bacteria prevents constipation and weight gain in mice, when compared to those fed a commercial formula milk.

INTRODUCCIÓN: Introducción: la leche materna (HM) contiene los nutrientes y compuestos bioactivos necesarios para el desarrollo infantil, incluidas bacterias probióticas, que contribuyen a la maduración intestinal. Objetivo: el objetivo de este estudio fue analizar la viabilidad de las bacterias acidolácticas aisladas de la leche humana con potencial probiótico, después del proceso de secado, así como evaluar su posible adhesión en el colón de ratones (BAlb/C) alimentados con leche humana en polvo y leche de una fórmula comercial. Método: se aislaron e identificaron mediante la técnica de Maldi-Tof-MS las bacterias acidolácticas con posible potencial probiótico en la leche humana en polvo. Se alimentó con leche humana en polvo y leche de una fórmula comercial a ratones de la cepa Bald/C durante 14 semanas. Se midieron el nivel de glucosa y el peso. Las heces se recolectaron para verificar la presencia de bacterias lácticas. Los ratones se sacrificaron y se pesaron los intestinos, aislando las bacterias lácticas tanto de los intestinos como de las heces. En las cepas aisladas de la leche humana se evaluó el potencial probiótico analizando su capacidad para inhibir patógenos, resistir distintos pH y temperaturas, adherirse y mostrar hidrofobicidad. Resultados: se identificó la presencia de Lactobacillus fermentum LH01, Lactobacillus rhamnosus LH02, Lactobacullis reuteri LH03 y L. plantarum LH05 en la leche humana en polvo. Todas las cepas mostraron resistencia a los pH bajos, a las sales biliares y a la exposición a enzimas gástricas, así como una buena hidrofobicidad y capacidad de autoagregación. Además, no presentaron actividad de hemaglutinación o hemólisis en un medio de cultivo rico en eritrocitos. Observamos que el consumo de leche humana en polvo evita en los ratones el aumento de peso y el estreñimiento. Conclusiones: después del secado por aspersión, las cepas con posible potencial probiótico pueden conservarse en la leche materna. El consumo de leche humana en polvo con bacterias probióticas evita el estreñimiento y el aumento de peso en los ratones, en comparación con los alimentados con leche de una formula comercial.

In-vivo assessment of the protection of Beta-glucans of Pleurotus ostreatus against oxidative stress caused by acrylamide intake (part II) / Evaluación in vivo de la proteccion de los beta-glucanos de Pleurotus ostreatus contra el estrés oxidativo causado por la ingesta de acrilamida (parte II)

INTRODUCTION: in April 2002, the National Food Authority of Sweden published a study in which the presence of a carcinogen was reported for the first time in experimental animals, and was identified as acrylamide. Various studies have shown that the β-glucans of Pleurotus ostreatus have diverse biological properties including antioxidant and anticancer activities. METHODS: β-glucans were obtained by alkaline-acid hydrolysis from Pleurotus ostreatus, and their content was characterized by liquid chromatography. To evaluate the effect of β-glucans on the expression of glutathione, Balb/c mice were used, and 4 test groups were established. All groups were fed as usual, groups treated with acrylamide were administered the compound intragastrically at a concentration of 50 μg/mL, and β-glucan treatment was given at a concentration of 50 μg/mL. RESULTS: no mortality was observed after exposure to the tested dose of acrylamide; only signs of peripheral neuropathy such as hyperactivity and tremors were observed after five days of experimentation, and were maintained over 30 days after the experiment. On the other hand, an increase in lipid peroxidation levels was observed in the livers of the acrylamide-treated mice, which were lower in the mice treated with β-glucans. CONCLUSIONS: results show that β-glucans may act as antioxidant agents able to protect the liver against oxidative stress as caused by the intake of acrylamide

INTRODUCCIÓN: en abril de 2002, la Autoridad Nacional de Alimentos de Suecia publicó un estudio en el que se informó por primera vez de la presencia de un carcinógeno en animales experimentales, identificado como acrilamida. Diversos estudios han demostrado que los β-glucanos de Pleurotus ostreatus tienen diversas propiedades biológicas, tales como actividades antioxidantes y anticancerígenas. MÉTODOS: los β-glucanos se obtuvieron por hidrólisis ácido-alcalina de Pleurotus ostreatus y su contenido se caracterizó por cromatografía líquida. Para evaluar el efecto de los β-glucanos sobre la expresión de glutatión, se usaron ratones Balb/c y se establecieron 4 grupos de prueba; todos los grupos se alimentaron normalmente, en los grupos tratados con acrilamida esta se administró intragástricamente a una concentración de 50 μg/mL, y el tratamiento con β-glucanos se dio a una concentración de 50 μg/mL. RESULTADOS: no se observó mortalidad después de la exposición a la dosis probada de acrilamida; solo se observaron signos de neuropatía periférica, como hiperactividad y temblores, después de cinco días de experimentación, que se mantuvieron dentro de los 30 días posteriores al experimento. Por otro lado, se observó un aumento de los niveles de peroxidación lipídica en los hígados de los ratones tratados con acrilamida, que fueron más bajos en los ratones tratados con β-glucanos. CONCLUSIONES: los resultados muestran que los β-glucanos podrían actuar como agentes antioxidantes y proteger el hígado contra el estrés oxidativo causado por la ingesta de acrilamida

In-vivo assessment of the protection of ß-glucans of Pleurotus ostreatus against oxidative stress caused by acrylamide intake (part II).

INTRODUCTION: Introduction: in April 2002, the National Food Authority of Sweden published a study in which the presence of a carcinogen was reported for the first time in experimental animals, and was identified as acrylamide. Various studies have shown that the ß-glucans of Pleurotus ostreatus have diverse biological properties including antioxidant and anticancer activities. Methods: ß-glucans were obtained by alkaline-acid hydrolysis from Pleurotus ostreatus, and their content was characterized by liquid chromatography. To evaluate the effect of ß-glucans on the expression of glutathione, Balb/c mice were used, and 4 test groups were established. All groups were fed as usual, groups treated with acrylamide were administered the compound intragastrically at a concentration of 50 g/mL, and ß-glucan treatment was given at a concentration of 50 g/mL. Results: no mortality was observed after exposure to the tested dose of acrylamide; only signs of peripheral neuropathy such as hyperactivity and tremors were observed after five days of experimentation, and were maintained over 30 days after the experiment. On the other hand, an increase in lipid peroxidation levels was observed in the livers of the acrylamide-treated mice, which were lower in the mice treated with ß-glucans. Conclusions: results show that ß-glucans may act as antioxidant agents able to protect the liver against oxidative stress as caused by the intake of acrylamide.

INTRODUCCIÓN: Introducción: en abril de 2002, la Autoridad Nacional de Alimentos de Suecia publicó un estudio en el que se informó por primera vez de la presencia de un carcinógeno en animales experimentales, identificado como acrilamida. Diversos estudios han demostrado que los ß-glucanos de Pleurotus ostreatus tienen diversas propiedades biológicas, tales como actividades antioxidantes y anticancerígenas. Métodos: los ß-glucanos se obtuvieron por hidrólisis ácido-alcalina de Pleurotus ostreatus y su contenido se caracterizó por cromatografía líquida. Para evaluar el efecto de los ß-glucanos sobre la expresión de glutatión, se usaron ratones Balb/c y se establecieron 4 grupos de prueba; todos los grupos se alimentaron normalmente, en los grupos tratados con acrilamida esta se administró intragástricamente a una concentración de 50 µg/mL, y el tratamiento con ß-glucanos se dio a una concentración de 50 µg/mL. Resultados: no se observó mortalidad después de la exposición a la dosis probada de acrilamida; solo se observaron signos de neuropatía periférica, como hiperactividad y temblores, después de cinco días de experimentación, que se mantuvieron dentro de los 30 días posteriores al experimento. Por otro lado, se observó un aumento de los niveles de peroxidación lipídica en los hígados de los ratones tratados con acrilamida, que fueron más bajos en los ratones tratados con ß-glucanos. Conclusiones: los resultados muestran que los ß-glucanos podrían actuar como agentes antioxidantes y proteger el hígado contra el estrés oxidativo causado por la ingesta de acrilamida.

In vivo assessment of the protection conferred by beta-glucans from Pleurotus ostreatus against the harmful effects of acrylamide intake (Part I) / Evaluación in vivo de la protección que confieren los beta-glucanos de Pleurotus ostreatus frente a los efectos nocivos de la ingestión de acrilamida (Parte I)

INTRODUCTION: acrylamide is formed in food through Maillard's reaction during thermal processing, and has been shown to be neurotoxic in humans, and a possible carcinogen. Studies have shown that β-glucans from Pleurotus ostreatus have diverse biological properties such as antioxidant and anticancer activities. OBJECTIVE: the aim of this work was to evaluate the protective effect of β-glucans from Pleurotus ostreatus against the harmful effects of acrylamide consumption in mice. METHODS: β-glucans were obtained by alkaline-acid hydrolysis of Pleurotus ostreatus, and the content was characterized by liquid chromatography. To evaluate the effect of β-glucans on the expression of glutathione, Balb/c mice were used, and 4 test groups were established. All groups were fed normally, and the groups treated with acrylamide were administered the compound intragastrically at a concentration of 50 μg/mL; β-glucans were administered at a concentration of 50 μg/mL. RESULTS: mice exposed to acrylamide showed a marked variation in the activity of glutathione enzymes in the liver. Significant differences (p < 0.05) were only found in the expression of glutathione transferase, which was increased almost 3 times in the group treated with β-glucans as compared with the control group, and 1.5 times as compared with the group treated with acrylamide. CONCLUSIONS: the results show that β-glucans could act by increasing the activity of enzymes involved in xenobiotic detoxification, thus protecting the biological system against the harmful effects caused by acrylamide intake

INTRODUCCIÓN: la acrilamida se forma en los alimentos a través de la reacción de Maillard durante el proceso térmico, y ha demostrado ser neurotóxica en humanos y un posible carcinógeno. Algunos estudios han demostrado que los β-glucanos de Pleurotus ostreatus tienen diversas propiedades biológicas, como actividades antioxidantes y anticancerígenas. OBJETIVO: el objetivo de este trabajo fue evaluar el efecto protector de los β-glucanos de Pleurotus ostreatus contra los efectos nocivos por consumo de acrilamida en ratones (prueba in vivo). MÉTODOS: los β-glucanos se obtuvieron por hidrólisis ácido-alcalina de Pleurotus ostreatus y su contenido se caracterizó por cromatografía líquida. La oxidación de los lípidos se evaluó mediante el método de TBARS, y para evaluar el efecto de los β-glucanos en la expresión de glutatión se usaron ratones Balb/c, y se establecieron 4 grupos de prueba. Todos los grupos fueron alimentados normalmente; a lo grupos tratados con acrilamida, esta se les administró intragástricamente en una concentración de 50 μg/ml, y los β-glucanos en una concentración de 50 μg/ml. RESULTADOS: en el presente trabajo, los ratones expuestos a acrilamida mostraron una marcada variación en la actividad de las enzimas de glutatión determinadas en el hígado. Solo se encontraron diferencias significativas (p < 0,05) en la expresión de glutatión-transferasa, que aumentó casi 3 veces en el grupo tratado con β-glucano en comparación con el grupo de control, y 1,5 veces con respecto al grupo tratado con acrilamida. CONCLUSIONES: los resultados muestran que los β-glucanos podrían actuar como agentes antioxidantes que protegen el hígado contra el estrés oxidativo causado por la ingesta de acrilamida

In vivo assessment of the protection conferred by ß-glucans from Pleurotus ostreatus against the harmful effects of acrylamide intake (Part I).

INTRODUCTION: Introduction: acrylamide is formed in food through Maillard's reaction during thermal processing, and has been shown to be neurotoxic in humans, and a possible carcinogen. Studies have shown that ß-glucans from Pleurotus ostreatus have diverse biological properties such as antioxidant and anticancer activities. Objective: the aim of this work was to evaluate the protective effect of ß-glucans from Pleurotus ostreatus against the harmful effects of acrylamide consumption in mice. Methods: ß-glucans were obtained by alkaline-acid hydrolysis of Pleurotus ostreatus, and the content was characterized by liquid chromatography. To evaluate the effect of ß-glucans on the expression of glutathione, Balb/c mice were used, and 4 test groups were established. All groups were fed normally, and the groups treated with acrylamide were administered the compound intragastrically at a concentration of 50 g/mL; ß-glucans were administered at a concentration of 50 g/mL. Results: mice exposed to acrylamide showed a marked variation in the activity of glutathione enzymes in the liver. Significant differences (p < 0.05) were only found in the expression of glutathione transferase, which was increased almost 3 times in the group treated with ß-glucans as compared with the control group, and 1.5 times as compared with the group treated with acrylamide. Conclusions: the results show that ß-glucans could act by increasing the activity of enzymes involved in xenobiotic detoxification, thus protecting the biological system against the harmful effects caused by acrylamide intake.

INTRODUCCIÓN: Introducción: la acrilamida se forma en los alimentos a través de la reacción de Maillard durante el proceso térmico, y ha demostrado ser neurotóxica en humanos y un posible carcinógeno. Algunos estudios han demostrado que los ß-glucanos de Pleurotus ostreatus tienen diversas propiedades biológicas, como actividades antioxidantes y anticancerígenas. Objetivo: el objetivo de este trabajo fue evaluar el efecto protector de los ß-glucanos de Pleurotus ostreatus contra los efectos nocivos por consumo de acrilamida en ratones (prueba in vivo). Métodos: los ß-glucanos se obtuvieron por hidrólisis ácido-alcalina de Pleurotus ostreatus y su contenido se caracterizó por cromatografía líquida. La oxidación de los lípidos se evaluó mediante el método de TBARS, y para evaluar el efecto de los ß-glucanos en la expresión de glutatión se usaron ratones Balb/c, y se establecieron 4 grupos de prueba. Todos los grupos fueron alimentados normalmente; a lo grupos tratados con acrilamida, esta se les administró intragástricamente en una concentración de 50 µg/ml, y los ß-glucanos en una concentración de 50 µg/ml. Resultados: en el presente trabajo, los ratones expuestos a acrilamida mostraron una marcada variación en la actividad de las enzimas de glutatión determinadas en el hígado. Solo se encontraron diferencias significativas (p < 0,05) en la expresión de glutatión-transferasa, que aumentó casi 3 veces en el grupo tratado con ß-glucano en comparación con el grupo de control, y 1,5 veces con respecto al grupo tratado con acrilamida. Conclusiones: los resultados muestran que los ß-glucanos podrían actuar como agentes antioxidantes que protegen el hígado contra el estrés oxidativo causado por la ingesta de acrilamida.

Downregulation of hippocampal NR2A/2B subunits related to cognitive impairment in a pristane-induced lupus BALB/c mice.

Neuropsychiatric systemic lupus erythematosus (NPSLE) is associated with learning and memory deficit. Murine model of lupus induced by pristane in BALB/c mice is an experimental model that resembles some clinical and immunological SLE pathogenesis. Nevertheless, there is no experimental evidence that relates this model to cognitive dysfunction associated with NR2A/2B relative expression. To evaluate cognitive impairment related to memory deficits in a murine model of lupus induced by pristane in BALB/c mice related to mRNA relative expression levels of NR2A/2B hippocampal subunits in short and long-term memory task at 7 and 12 weeks after LPS exposition in a behavioral test with the use of Barnes maze. A total of 54 female BALB/c mice 8-12 weeks old were included into 3 groups: 7 and 12 weeks using pristane alone (0.5 mL of pristane) by a single intraperitoneal (i.p.) injection. A control group (single i.p. injection of 0.5 mL NaCl 0.9%) and pristane plus LPS exposure using single i.p. pristane injection and LPS of E. coli O55:B5, in a dose of 3mg/kg diluted in NaCl 0.9% 16 weeks post-pristane administration. To determine cognitive dysfunction, mice were tested in a Barnes maze. Serum anti-Sm antibodies and relative expression of hippocampal NR2A/2B subunits (GAPDH as housekeeping gene) with SYBR green quantitative reverse transcription polymerase chain reaction and 2-ΔΔCT method were determined in the groups. Downregulation of hippocampal NR2A subunit was more evident than NR2B in pristane and pristane+LPS at 7 and 12 weeks of treatment and it is related to learning and memory disturbance assayed by Barnes maze. This is the first report using the murine model of lupus induced by pristane that analyzes the NMDA subunit receptors, finding a downregulation of NR2A subunit related to learning and memory disturbance being more evident when they were exposed to LPS.

Potential Chronotherapeutic Optimization of Antimalarials in Systemic Lupus Erythematosus: Is Toll-Like Receptor 9 Expression Dependent on the Circadian Cycle in Humans?

Toll-like receptor 9 (TLR9) belongs to the group of endosomal receptors of the innate immune system with the ability to recognize hypomethylated CpG sequences from DNA. There is scarce information about TLR9 expression and its association with the circadian cycle (CC). Different patterns of TLR9 expression are regulated by the CC in mice, with an elevated expression at Zeitgeber time 19 (1:00 a.m.); nevertheless, we still need to corroborate this in humans. In systemic lupus erythematosus (SLE), the inhibitory effect of chloroquine (CQ) on TLR9 is limited. TLR9 activation has been associated with the presence of some autoantibodies: anti-Sm/RNP, anti-histone, anti-Ro, anti-La, and anti-double-stranded DNA. Treatment with CQ for SLE has been proven to be useful, in part by interfering with HLA-antigen coupling and with TLR9 ligand recognition. Studies have shown that TLR9 inhibitors such as antimalarial drugs are able to mask TLR9-binding sites on nucleic acids. The data presented here provide the basic information that could be useful for other clinical researchers to design studies that will have an impact in achieving a chronotherapeutic effect by defining the ideal time for CQ administration in SLE patients, consequently reducing the pathological effects that follow the activation of TLR9.

Low CD36 and LOX-1 Levels and CD36 Gene Subexpression Are Associated with Metabolic Dysregulation in Older Individuals with Abdominal Obesity.

Background. Obesity study in the context of scavenger receptors has been linked to atherosclerosis. CD36 and LOX-1 are important, since they have been associated with atherogenic and metabolic disease but not fat redistribution. The aim of our study was to determinate the association between CD36 and LOX-1 in presence of age and abdominal obesity. Methods. This is a cross-sectional study that included 151 healthy individuals, clinically and anthropometrically classified into two groups by age (<30 and ≥30 years old) and abdominal obesity (according to World Health Organization guidelines). We excluded individuals with any chronic and metabolic illness, use of medication, or smoking. Fasting blood samples were taken to perform determination of CD36 mRNA expression by real-time PCR, lipid profile and metabolic and low grade inflammation markers by routine methods, and soluble scavenger receptors (CD36 and LOX-1) by ELISA. Results. Individuals ≥30 years old with abdominal obesity presented high atherogenic index, lower soluble scavenger receptor levels, and subexpression of CD36 mRNA (54% less). On the other hand, individuals <30 years old with abdominal adiposity presented higher levels in the same parameters, except LOX-1 soluble levels. Conclusion. In this study, individuals over 30 years of age presented low soluble scavenger receptors levels pattern and CD36 gene subexpression, which suggest the chronic metabolic dysregulation in abdominal obesity.

CCL2 Serum Levels and Adiposity Are Associated with the Polymorphic Phenotypes -2518A on CCL2 and 64ILE on CCR2 in a Mexican Population with Insulin Resistance.

Genetic susceptibility has been described in insulin resistance (IR). Chemokine (C-C motif) ligand-2 (CCL2) is overexpressed in white adipose tissue and is the ligand of C-C motif receptor-2 (CCR2). The CCL2 G-2518A polymorphism is known to regulate gene expression, whereas the physiological effects of the CCR2Val64Ile polymorphism are unknown. The aim of the study is to investigate the relationship between these polymorphisms with soluble CCL2 levels (sCCL2), metabolic markers, and adiposity. In a cross-sectional study we included 380 Mexican-Mestizo individuals, classified with IR according to Stern criteria. Polymorphism was identified using PCR-RFLP/sequence-specific primers. Anthropometrics and metabolic markers were measured by routine methods and adipokines and sCCL2 by ELISA. The CCL2 polymorphism was associated with IR (polymorphic A+ phenotype frequencies were 70.9%, 82.6%, in individuals with and without IR, resp.). Phenotype carriers CCL2 (A+) displayed lower body mass and fat indexes, insulin and HOMA-IR, and higher adiponectin levels. Individuals with IR presented higher sCCL2 compared to individuals without IR and was associated with CCR2 (Ile+) phenotype. The double-polymorphic phenotype carriers (A+/Ile+) exhibited higher sCCL2 than double-wild-type phenotype carriers (A-/Ile-). The present findings suggest that sCCL2 production possibly will be associated with the adiposity and polymorphic phenotypes of CCL2 and CCR2, in Mexican-Mestizos with IR.

Rapid compensatory changes in the expression of EAAT-3 and GAT-1 transporters during seizures in cells of the CA1 and dentate gyrus.

BACKGROUND: Epilepsy is a neurological disorder produced by an imbalance between excitatory and inhibitory neurotransmission, in which transporters of both glutamate and GABA have been implicated. Hence, at different times after local administration of the convulsive drug 4-aminopyridine (4-AP) we analyzed the expression of EAAT-3 and GAT-1 transporter proteins in cells of the CA1 and dentate gyrus. METHODS: Dual immunofluorescence was used to detect the co-localization of transporters and a neuronal marker. In parallel, EEG recordings were performed and convulsive behavior was rated using a modified Racine Scale. RESULTS: By 60 min after 4-AP injection, EAAT-3/NeuN co-labelling had increased in dentate granule cells and decreased in CA1 pyramidal cells. In the latter, this decrease persisted for up to 180 min after 4-AP administration. In both the DG and CA1, the number of GAT-1 labeled cells increased 60 min after 4-AP administration, although by 180 min GAT-1 labeled cells decreased in the DG alone. The increase in EAAT-3/NeuN colabelling in DG was correlated with maximum epileptiform activity and convulsive behavior. CONCLUSIONS: These findings suggest that a compensatory mechanism exists to protect against acute seizures induced by 4-AP, whereby EAAT-3/NeuN cells is rapidly up regulated in order to enhance the removal of glutamate from the extrasynaptic space, and attenuating seizure activity.

Effect of early glutamate exposure on EAAT-3 and GAT-1 protein expression in cells of the dentate gyrus and CA1 region of the adult rat hippocampus.

BACKGROUND AND AIMS: Glutamate and GABA transporters are cell surface proteins localized on neurons and glial cells that mediate the reuptake of glutamate and GABA from the extracellular space. In different models of the acquisition of epilepsy, important changes in the expression of these transporters have been demonstrated, although to date no such studies have been performed using the monosodium glutamate (MSG)-induced seizure model in neonatal rats. METHODS: Following repeated MSG administration, we used immunofluorescence techniques to quantify the number of cells expressing the EAAT-3 and GAT-1 transporters at postnatal days (PD) 14 and 60 in the dentate gyrus (DG) and CA1 region of the hippocampus. RESULTS: EAAT-3 and GAT-1 were expressed around the soma of granular cells and in the soma and dendrites of pyramidal cells in both experimental (MSG) and control (NaCl) rats. In the DG, MSG administration significantly increased the number of granular cells double-labelled for EAAT-3/Neun at PD 60 but not PD 14. No significant changes were observed at either PD 14 or 60 in terms of the number of cells expressing GAT-1 in the DG or CA1. CONCLUSIONS: The findings suggest that the selective long-term increase in EAAT-3 expression in granular cells following neonatal MSG treatment reflects an important compensatory or protective response to the excitotoxic and seizure-promoting effects of early glutamate exposure in adult animals.