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BACKGROUND: The aim was to diagnose Candida in the oral cavity of subjects with type 2 diabetes mellitus (T2DM) using a genotyping technique and compare the results with those from conventional diagnosis by Papanicolaou (Pap) staining. METHODS: Palatal mucosa smears were performed on 18 dental care patients diagnosed with T2DM and grade I, II, and III prosthetic stomatitis who met the inclusion criteria; 18 healthy control subjects were also included in the study. Hemoglobin A1c (HbA1c) levels were determined from total blood. Using exfoliative cytology, the Pap staining technique was used to diagnose candidiasis. Exfoliative cytology was also used for molecular diagnosis; DNA was obtained for Candida genotyping, and RNA was used for gene expression studies. RESULTS: Clinical patterns indicated that all subjects were positive for Candida; however, Pap analysis revealed only three positive subjects, whereas end-point polymerase chain reaction (PCR) analysis revealed 15 subjects with some type of Candida. The most common Candida species found were Candida guilliermondii (38.8%), Candida krusei (33.3%), Candida tropicalis, and Candida lusitaniae (22.2%). Interestingly, the coexpression of different species of Candida was found in various patients. In all patients, HbA1c levels were increased. Gene expression analysis showed a significant decrease (p ≤ 0.05) in TLR2 expression in positive subjects, whereas TLR4 expression did not differ significantly among patients. CONCLUSIONS: The end-point PCR technique showed better sensitivity for the diagnosis of Candida when compared with the diagnosis by Pap staining. T2DM subjects showed an increased presence of C. guilliermondii that was correlated with decreased TLR2 expression.
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Resumen Antecedentes: La sospecha de disfunción endotelial (DE) permitirá prevenir la aterosclerosis acelerada y la muerte prematura. Objetivo: Establecer la utilidad de la termografía en el cribado de la función endotelial en adultos con factores de riesgo cardiovascular. Material y métodos: Estudio transversal analítico de prueba diagnóstica. El incremento del diámetro de la arteria braquial < 11 % a un minuto posisquemia significó probable DE, confirmada si el diámetro fue ≥ 11 % posnitroglicerina sublingual. Se obtuvieron fotografías termográficas al minuto de la región palmar. Se aplicó estadística descriptiva, curva ROC, pruebas U de Mann-Whitney, chi cuadrada o exacta de Fisher. Resultados: Se incluyeron 38 sujetos, mediana de edad de 50 años, con 624 mediciones termográficas; nueve presentaron DE (vasodilatación mediada por flujo de 2.5 %). El mejor punto de corte para la función endotelial normal en sujetos con factores de riesgo cardiovascular fue ≥ 36 °C al minuto de isquemia, con sensibilidad de 85%, especificidad de 70%, valores predictivos positivo y negativo de 78 y 77%, área bajo la curva de 0.796, razón de verisimilitud positiva de 2.82 y razón de verisimilitud negativa de 0.22. Conclusión: La medición de la temperatura en la región palmar mediante termografía infrarroja ≥ 36 °C tras un minuto de isquemia es práctica, no invasiva y económica para el cribado de la función endotelial normal en adultos con factores de riesgo cardiovascular.
Abstract Background: Endothelial dysfunction (ED) suspicion will allow to prevent accelerated atherosclerosis and premature death. Objective: To establish the usefulness of thermography for endothelial function screening in adults with cardiovascular risk factors. Material and methods: Cross-sectional, analytical diagnostic test. A brachial arterial diameter (BAD) increase <11 % at one-minute post-ischemia meant probable ED and was confirmed if BAD was ≥ 11 % post-sublingual nitroglycerin. Thermographic photographs of the palmar region were obtained at one minute. Descriptive statistics, ROC curve, Mann-Whitneys U-test, chi-square test, or Fishers exact test were used. Results: Thirty-eight subjects with a median age of 50 years, and with 624 thermographic measurements were included. Nine had ED (flow-mediated vasodilation (FMV): 2.5 %. The best cutoff point for normal endothelial function in subjects with cardiovascular risk factors was ≥ 36 °C at one minute of ischemia, with 85 % sensitivity, 70 % specificity, positive and negative predictive values of 78 and 77 %, area under the curve of 0.796, LR+ 2.82, LR- 0.22. Conclusions: An infrared thermography-measured temperature in the palmar region greater than or equal to 36 °C after one minute of ischemia is practical, non-invasive, and inexpensive for normal endothelial function screening in adults with cardiovascular risk factors.
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Bioactive peptides are biomolecules involved in very diverse mechanisms in vivo. It has been reported that bioactive peptides play a very important role in the regulation of physiological functions such as oxidative stress, hypertension, cancer and inflammation. It's been reported that the milk derived peptide (VPP) prevents the progress of hypertension in different animal models and human beings with mild hypertension. It has also been shown that oral administration of VPP produces an anti-inflammatory effect in adipose tissue of mouse models. Currently there are no reports on the possible interaction of VPP with the enzymes superoxide dismutase (SOD) and catalase (CAT), the main regulators of oxidative stress. This study analyzes the interaction between VPP and specific domains in the minimal promoter region of the genes SOD and CAT in blood samples of obese children using a QCM-D type piezoelectric biosensor. We also used molecular modeling (docking) to determine the interaction between the peptide VPP and the minimal promoter region of both genes. With QCM-D, we detected the interaction of VPP with the nitrogenous base sequences that comprise the minimal promoter regions of both genes CAT and SOD. These experimental interactions were explained at the atomic level by molecular docking simulations showing how the peptides are capable of reaching the DNA structures by means of hydrogen bonds with favored free energy values. It is possible to conclude that the combined use of docking and QCM-D allows for the determination of the interaction of small peptides (VPP) with specific sequences of genes.
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Hipertensão , Obesidade Infantil , Criança , Camundongos , Animais , Humanos , Catalase/genética , Simulação de Acoplamento Molecular , Peptídeos/genética , Superóxido Dismutase/genética , Regiões Promotoras Genéticas/genéticaRESUMO
AIM: The purpose of this work was to identify and measure catecholamines, their metabolites, and the gene expression of catecholamine receptors in osteosarcoma tissue. MATERIALS AND METHODS: The levels of 3,4-dihydroxyphenylacetic acid, norepinephrine, serotonin, and 5-hydroxyindoleacetic acid in cancer tissue and in adjacent and non-oncological bone tissue were analysed by high-performance liquid chromatography, and the gene expression of catecholamine receptors and of dopamine ß-hydroxylase, monoaminoxidase, ki67, and Runx2 in the osteosarcoma tissue, tissue adjacent to the tumour, non-oncological bone, and human brain tissue was analysed by RT-PCR. RESULTS: We found significantly higher levels of 3,4-dihydroxyphenylacetic acid and norepinephrine in the cancer sample than in adjacent and non-oncological bone. We found that ß-adrenergic receptors and dopaminergic receptors, dopamine ß-hydroxylase, ki67, Runx2, and serotonergic receptor gene expression were significantly higher in tumour tissue than in adjacent and non-oncological bone. CONCLUSION: Catecholamines and their receptors could be potential molecular markers for osteosarcoma progression.
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Neoplasias Ósseas/patologia , Catecolaminas/metabolismo , Regulação da Expressão Gênica , Metaboloma , Osteossarcoma/patologia , Receptores de Catecolaminas/metabolismo , Idoso , Neoplasias Ósseas/genética , Neoplasias Ósseas/metabolismo , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Osteossarcoma/genética , Osteossarcoma/metabolismo , Receptores de Catecolaminas/genéticaRESUMO
Resumen Introducción: El food craving o ansia por comer es una respuesta motivacional y fisiológica por comer alimentos específicos, principalmente con alto contenido calórico. Para evaluarlo se usa, entre otros, el Food Craving Questionnaire Trait, estructurado multidimensionalmente y validado en diversos países, el cual ha mostrado ser sensible y adaptable a los cambios contextuales-culturales. Objetivos: Validar y estandarizar el Food Craving Questionnaire-Trait en adultos de la Ciudad de México. Método: Estudio no experimental, transversal y aleatorizado de 1059 sujetos de uno y otro sexo, entre 18 y 84 años; 71.86 % del sexo femenino. Se examinaron propiedades psicométricas con análisis factoriales exploratorios y confirmatorios. Resultados: Se redujeron los factores del cuestionario y los ítems se reorganizaron de forma diferente al original. El análisis factorial confirmatorio mostró ajuste adecuado y estandarización aceptable de los factores. Se encontró alta consistencia interna para el cuestionario global (a = 0.973 y rho = 0.975) para cada uno de los factores. Conclusión: Este estudio determina la viabilidad del Food Craving Questionnaire para población de la Ciudad de México.
Abstract Introduction: Food craving is a motivational and physiological response for eating specific foods, mainly with high caloric content. To assess food craving, self-reports, inventories and questionnaires are used. The Food Cravings Questionnaire-Trait is multi-dimensionally structured and has been validated in several countries, since it is sensitive and adaptable to contextual-cultural changes. Objectives: To validate and standardize the Food Cravings Questionnaire-Trait in adults of Mexico City. Method: Non-experimental, cross-sectional, randomized study of 1059 subjects of both genders, between 18 and 84 years of age; 71.86 % of the female gender. Psychometric properties were examined with exploratory and confirmatory factor analyses. Results: The domains of the questionnaire were reduced and the items were reorganized differently from the original version. The confirmatory factor analysis showed an adequate fit and acceptable standardization of factors. High internal consistency was found for the global questionnaire (a = 0.973 and rho = 0.975) for each one of the domains. Conclusion: This study determines the viability of the Food Cravings Questionnaire for the population of Mexico City.
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Humanos , Masculino , Adolescente , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Adulto Jovem , Apetite/fisiologia , Inquéritos e Questionários/normas , Fissura/fisiologia , Alimentos , Psicometria , Estudos Transversais , Reprodutibilidade dos Testes , Emoções , Comportamento Alimentar , Antecipação Psicológica , Culpa , Comportamento Impulsivo/fisiologia , MéxicoRESUMO
Alzheimer's disease (AD) is one of the most complicated neurodegenerative diseases, and several hypotheses have been associated with its development and progression, such as those involving glucose hypometabolism, the cholinergic system, calcium imbalance, inflammation, oxidative imbalance, microtubule instability, and the amyloid cascade, several of which are related to oxidative stress (free radical generation), which contributes to neuronal death. Therefore, several efforts have been made to establish a sporadic AD model that takes into account these hypotheses. One model that replicates the increase in amyloid beta (Aß) and oxidative stress in vivo is the scopolamine model. In the present work, the chronic administration (6 weeks) of scopolamine was used to analyze the neuroprotective effects of apocynin and galantamine. The results showed that scopolamine induced cognitive impairment, which was evaluated 24 h after the final dose was administered. In addition, after scopolamine administration, the Aß and superoxide anion levels were increased, and NADPH oxidase 2 (NOX2), nuclear factor erythroid 2-related factor 2 (Nrf2), and nuclear factor kappa B (NFkB) genes were overexpressed. These effects were not observed when either apocynin or galantamine was administered during the last 3 weeks of scopolamine treatment, and although the results from both molecules were related to lower Aß production and, consequently, lower superoxide anion production, they were likely realized through different pathways. That is, both apocynin and galantamine diminished NADPH oxidase expression, but their effects on transcription factor expression differed. Moreover, experiments in silico showed that galantamine did not interact with the active site of beta secretase, whereas diapocynin, an apocynin metabolite, interacted with the beta-site APP-cleaving enzyme (BACE1) at the catalytic site.
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Acetofenonas/uso terapêutico , Doença de Alzheimer/tratamento farmacológico , Galantamina/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Acetofenonas/farmacologia , Doença de Alzheimer/etiologia , Peptídeos beta-Amiloides/metabolismo , Animais , Cognição , Galantamina/farmacologia , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Masculino , NADPH Oxidase 2/genética , NADPH Oxidase 2/metabolismo , NF-kappa B/genética , NF-kappa B/metabolismo , Fármacos Neuroprotetores/farmacologia , Estresse Oxidativo , Ratos , Ratos Wistar , Escopolamina/toxicidadeRESUMO
BACKGROUND: In microsurgical reconstruction, vascular obstruction occurs in approximately 20% of patients. Close monitoring is central to their care. Clinical/Doppler detection of vascular obstruction could be enhanced by thermography. METHODS: A diagnostic test design included consecutive cases of hospitalized patients, ≥18 years old, who underwent surgery with free flaps. Two researchers separately evaluated patients with clinical/Doppler methods and thermographic camera hourly for 24 hours, every 2 hours for the next 24 hours, and then every 3 hours until discharge. The gold standard was visualization of thrombus or vascular obstruction during surgical reintervention. Sensitivity, specificity, positive/negative predictive value (PPV/NPV), and a delta temperature receiver operating characteristic (ROC) curve were calculated. RESULTS: A total of 2,364 tests were performed with a thermographic camera in 40 patients (31 females, 9 males) aged 50.12 ± 9.7 years. There were 28 deep inferior epigastric perforator, 5 anterolateral thigh, 3 radial, 2 scapular, 1 fibular, and 1 anteromedial thigh flaps included. Six (15%) had postoperative vascular obstruction (5 venous and 1 arterial). One flap developed partial necrosis and one total necrosis (overall survival 97.5%). ROC curve (area 0.97) showed the best results at ≥ 1.8°C of difference to the surrounding skin. Considering two consecutive positive evaluations, the sensitivity was 93%, specificity 96%, PPV 57%, and NPV 99%. The thermal imaging camera allows to identify the obstruction between 2 and 12 hours before the clinical method. CONCLUSION: Utilizing a thermographic camera can reduce detection time of vascular obstruction by several hours in microvascular free flaps that include the cutaneous island. This method proves useful for early diagnosis of postoperative vascular obstruction.
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Retalhos de Tecido Biológico/irrigação sanguínea , Oclusão de Enxerto Vascular/diagnóstico , Termografia/instrumentação , Diagnóstico Precoce , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Sensibilidade e EspecificidadeRESUMO
Brown algae and its carotenoids have been shown to have a positive influence on obesity and its comorbidities. This study evaluated the effect of Undaria pinnatifida and fucoxanthin on biochemical, physiological and inflammation markers related to obesity and on the expression of genes engaged on white adipose tissue lipid metabolism in a murine model of diet-induced obesity. The treatments improved energy expenditure, ß-oxidation and adipogenesis by upregulating PPARα, PGC1α, PPARγ and UCP-1. Adipogenesis was also confirmed by image analysis of the retroperitoneal adipose tissue, by measuring cell area, perimeter and cellular density. Additionally, the treatments, ameliorated adipose tissue accumulation, insulin resistance, blood pressure, cholesterol and triglycerides concentration in serum, and reduced lipogenesis and inflammation by downregulating acetyl-CoA carboxylase (ACC) gene expression, increasing serum concentration and expression of adiponectin as well as downregulating IL-6 expression. Both fucoxanthin and Undaria pinnatifida may be considered for treating obesity and other diseases related.
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Dieta Vegetariana/métodos , Lipogênese/efeitos dos fármacos , Obesidade/dietoterapia , Phaeophyceae/química , Undaria/química , Xantofilas/farmacologia , Acetil-CoA Carboxilase/metabolismo , Adiponectina/sangue , Adiponectina/metabolismo , Tecido Adiposo Branco/metabolismo , Animais , Biomarcadores/metabolismo , Dieta Hiperlipídica/efeitos adversos , Modelos Animais de Doenças , Humanos , Inflamação/dietoterapia , Interleucina-6/metabolismo , Masculino , Síndrome Metabólica/dietoterapia , Obesidade/etiologia , PPAR alfa/metabolismo , PPAR gama/metabolismo , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Ratos , Ratos Wistar , Proteína Desacopladora 1/metabolismo , Xantofilas/uso terapêuticoRESUMO
OBJECTIVE: Malnutrition has been associated with oxidative damage by altered antioxidant protection mechanisms. Specifically, the aim of this study was to evaluate oxidative damage (DNA and lipid) and antioxidant status (superoxide dismutase [SOD], glutathione peroxidase [GPx], and catalase [CAT] mRNA, and protein expression) in thymus from malnourished rat pups. METHODS: Malnutrition was induced during the lactation period by the food competition method. Oxidative DNA damage was determined quantifying 8-oxo-7, 8-dihydro-2'-deoxyguanosine adduct by high-performance liquid chromatography. Lipid peroxidation was assessed by the formation of thiobarbituric acid-reactive substances. Levels of gene and protein expression of SOD, GPx, and CAT were evaluated by real-time polymerase chain reaction and Western blot, respectively. Antioxidant enzyme activities were measured spectrophotometrically. RESULTS: Oxidative DNA damage and lipid peroxidation significantly increased in second-degree (MN-2) and third-degree malnourished (MN-3) rats compared with well-nourished rats. Higher amounts of oxidative damage, lower mRNA expression, and lower relative concentrations of protein, as well as decreased antioxidant activity of SOD, GPx, and CAT were associated with the MN-2 and MN-3 groups. CONCLUSIONS: The results of this study demonstrated that higher body-weight deficits were related to alterations in antioxidant protection, which contribute to increased levels of damage in the thymus. To our knowledge, this study demonstrated for the first time that early in life, malnutrition leads to increased DNA and lipid oxidative damage, attributable to damaged antioxidant mechanisms including transcriptional and enzymatic activity alterations. These findings may contribute to the elucidation of the causes of previously reported thymus dysfunction, and might explain partially why children and adults who have overcome child undernourishment experience immunologic deficiencies.
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Antioxidantes/metabolismo , Lactação , Desnutrição/metabolismo , Estado Nutricional , Estresse Oxidativo , Oxirredutases/metabolismo , Timo/metabolismo , Animais , Peso Corporal , Catalase/metabolismo , Dano ao DNA , Feminino , Glutationa Peroxidase/metabolismo , Peroxidação de Lipídeos , Oxirredução , RNA Mensageiro/metabolismo , Ratos Wistar , Superóxido Dismutase/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
CONTEXT: It has been reported that 17ß-estradiol (E2) reduces the expression of inflammatory molecules, but there are no data that show the effect of E2 on the transcriptional regulation of innate immunity-related molecules and inflammasomes. OBJECTIVE: To study the effect of 17ß-estradiol (E2) on the transcriptional expression of the NLR family, pyrin domain containing 1 (Nlrp1) and (Nlrp3) inflammasomes, which are mediators of inflammation. MATERIALS AND METHODS: Inflammation was induced in adult female gonadectomized (Gdx) rats by intramuscular injection of complete Freund's adjuvant (CFA). Measurements were taken at different times after the treatment. Gene expression determinations were done by quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: CFA-induced inflammation increased the transcription of Nlrp3, IL-1ß (p < 0.05), vascular cell adhesion molecule 1 (VCAM1), E-selectin and estrogen receptor 1 alpha (ERα) (p < 0.001) and decreased the transcription of Nlrp1, Caspase-1, IL-33, NFKB1, ICAM1, ICAM2, GCRα, GCRß, UCP3 and PGC1α (p < 0.001) compared to the control. The administration of E2 to the inflamed tissue significantly increased the expression of Nlrp1, NFKB1, ERα, UCP3, Caspase-1, E-selectin (p < 0.001), IL-18 and ERα (p < 0.05) and decreased IL-1ß and VCAM1 (p < 0.005) compared to the control. DISCUSSION AND CONCLUSION: CFA differentially modulates the transcription of inflammasome-related genes and the administration of E2 increases the expression of ERα and Nlrp1 together with NFKB1, a key molecule in the activation of the inflammasomes. Finally, an analysis using the web interface GeneMANIA revealed an interaction between several genes, indicating a functional correlation in this model.
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Proteínas de Transporte/genética , Estradiol/farmacologia , Imunidade Inata/efeitos dos fármacos , Inflamassomos/genética , Proteínas do Tecido Nervoso/genética , Transcrição Gênica/efeitos dos fármacos , Adjuvantes Imunológicos/administração & dosagem , Animais , Sequência de Bases , Edema/induzido quimicamente , Edema/imunologia , Feminino , Adjuvante de Freund/administração & dosagem , Adjuvante de Freund/imunologia , Inflamassomos/imunologia , Dados de Sequência Molecular , Proteína 3 que Contém Domínio de Pirina da Família NLR , Ovariectomia , Ratos WistarRESUMO
BACKGROUND: The activated NLRP3 inflammasome is associated with the etiology of fibrotic diseases. The role of inflammasomes in SSc is still poorly understood. OBJECTIVES: To determine the expression of NLRP3 (nucleotide-binding domain, leucine-rich-repeat-containing family, pyrin domain-containing 3) in the skin of patients with systemic sclerosis (SSc) and its relationship with pro-inflammatory cytokines and vascular mediators expression. METHODS: Skin biopsies were taken from 42 patients with either limited or diffuse SSc (21 lcSSc and 21 dcSSc), and from 13 healthy individuals. Using real-time polymerase chain reaction (PCR), the relative expression of caspase-1, IL-1ß, IL-18, IL-33, TGF-ß, ET-1, iNOS and eNOS genes, were measured. The location of NLRP3 and IL-1ß were also determined by immunohistochemistry. Clinical characteristics were evaluated. RESULTS: The mean age of the patients was 49.3 ± 12.9 (lcSSc), 44.6 ± 1 3.8 (dcSSc), and 45 ± 14.1 (healthy individuals). Compared to healthy individuals, the skin of both subtypes of SSc showed a significant increase (P < 0.05) in NLRP3, caspase-1, IL-1ß, IL-18 and ET-1. Samples of lcSSc also showed a significant increase of eNOS (P < 0.029), iNOS (P < 0.04) and TGF-ß (P < 0.05). Dermal fibrosis evaluated by modified Rodnan skin score (MRSS) had significant correlation with NLRP3, IL-1ß, IL-18, and ET-1. Immunohistochemical analysis showed stronger staining of NLRP3 and IL-1ß cytoplasmic expression in the keratinizing squamous epithelium of skin from SSc patients compared to controls. CONCLUSIONS: This study identified NLRP3 over-expression in skin of patients with SSc. Skin thickness correlates positively with the NLRP3 inflammasome gene expression and with the vascular mediator and pro-fibrotic ET-1, suggesting that NLRP3 inflammasome plays a role in the pathophysiology of skin fibrosis in human SSc.
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Proteínas de Transporte/genética , Citocinas/metabolismo , Inflamassomos/metabolismo , Escleroderma Sistêmico/patologia , Pele/patologia , Adulto , Biópsia , Endotelina-1/metabolismo , Feminino , Fibrose , Regulação da Expressão Gênica , Humanos , Mediadores da Inflamação/metabolismo , Masculino , Pessoa de Meia-Idade , Proteína 3 que Contém Domínio de Pirina da Família NLR , Reação em Cadeia da Polimerase em Tempo Real , Escleroderma Sistêmico/genética , Escleroderma Sistêmico/imunologiaRESUMO
Stress stimuli affect the immune system of the mucosa, and in particular IgA secretion. It is well documented that intense psychological and physical stress can increase susceptibility to infection by diverse pathogens in the upper respiratory tract. Our workgroup reported that chronic stress caused by immobilization elicits a decrease in nasal IgA levels in mice. Here, we explore how acute stress (caused by 4h of immobilization) affects IgA secretion in the nasal mucosa, and the possible role of the sympathetic nervous system in this effect. Nine-week-old male CD1 mice were divided into four groups: control, chemical sympathectomy (with 6-OHDA) and treatment with nadolol (5mg/kg) or phentolamine (15mg/kg). All these groups were subdivided into stressed and unstressed animals. The parameters evaluated included plasma corticosterone and epinephrine (only in control groups), SIgA levels (by ELISA) and SIgA expression (by Western Blot) in nasal fluid, percentage of IgA+ plasma cells, and mRNA expression of heavy alpha chain, pIgR, TNFα and TGFß in nasal mucosa. Acute stress reduced the percentage of IgA+ cells while increasing the levels of IgA, the two hormones, and the mRNA expression of heavy alpha chain, pIgR, TNFα and TGFß, which resulted in greater synthesis and transport of IgA. The treatments with 6-OHDA and α- and ß-adrenergic receptor blockers suggest that sympathetic innervation by both types of adrenergic receptors is important for the control of SIgA secretion in nasal mucosa during acute stress. The increase in this parameter depended on the cytokines involved in IgA synthesis and transport.
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Catecolaminas/metabolismo , Imunoglobulina A/metabolismo , Mucosa/metabolismo , Estresse Psicológico/metabolismo , Antagonistas Adrenérgicos alfa , Antagonistas Adrenérgicos beta/farmacologia , Animais , Corticosterona , Citocinas/metabolismo , Modelos Animais de Doenças , Imunoglobulina A Secretora/metabolismo , Masculino , Camundongos , Mucosa/efeitos dos fármacos , Nadolol/farmacologia , Oxidopamina/farmacologia , Fentolamina/farmacologia , RNA Mensageiro/metabolismo , Simpatectomia Química , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Cadmium (Cd(2+)) produces toxic effects on various tissues as kidney and liver, so several studies have focused to explore the effect produced by different doses and exposure times of this metal. However, little has been reported about the effect that Cd(2+) shows in the brain in vivo. Hence, this study aimed at comparing the effect of chronic Cd(2+) exposure on antioxidant defense systems of kidney and brain in rats. Six groups of male rats were employed; five were administered for 45 days with different doses of cadmium chloride (0.187, 0.375, 0.562, 0.937 and 1.125 mg/kg; i.p.) and the other was used as control. Free radicals (FRs) were directly quantified by electron paramagnetic resonance (EPR) spectroscopy; malondialdehyde (MDA), reduced glutathione (GSH) and the activity expression of superoxide dismutase (SOD2) and catalase (CAT) were also measured. The EPR results showed that there was no increase in FR content in kidney or brain. MDA and GSH levels increased in kidney but not in the brain. The SOD2 activity was not altered, but its expression decreased in both tissues. On the other hand, CAT activity and expression tended to increase at low doses and decrease at high doses in both tissues. Therefore, these results suggest that there exist compensatory mechanisms in both kidney and brain that are capable of avoiding the toxic effects exerted by Cd(2+) at these doses and exposure time.
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Antioxidantes/metabolismo , Encéfalo/efeitos dos fármacos , Cádmio/toxicidade , Rim/efeitos dos fármacos , Animais , Encéfalo/enzimologia , Encéfalo/metabolismo , Cádmio/administração & dosagem , Catalase/metabolismo , Espectroscopia de Ressonância de Spin Eletrônica , Glutationa/metabolismo , Rim/enzimologia , Rim/metabolismo , Masculino , Malondialdeído/metabolismo , Ratos , Ratos Wistar , Superóxido Dismutase/metabolismoRESUMO
Although caloric restriction (CR) apparently has beneficial effects on the immune system, its effects on the immunological function of the intestinal mucosa are little known. The present study explored the effect of CR on the innate and adaptive intestinal immunity of mice. Balb/c mice were either fed ad libitum (control) or on alternate days fed ad libitum and fasted (caloric restriction). After 4 months, an evaluation was made of IgA levels in the ileum, the gene expression for IgA and its receptor (pIgR), as well as the expression of two antimicrobial enzymes (lysozyme and phospholipase A2) and several cytokines of the intestinal mucosa. CR increased the gene expression of lysozyme and phospholipase A2. The levels of IgA were diminished in the ileum, which apparently was a consequence of the reduced transport of IgA by pIgR. In ileum, CR increased the gene expression for most cytokines, both pro- and anti-inflammatory. Hence, CR differentially modified the expression of innate and adaptive immunity mediators in the intestine.
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Imunidade Adaptativa , Restrição Calórica , Duodeno/imunologia , Íleo/imunologia , Imunidade Inata , Animais , Peso Corporal , Corticosterona/sangue , Citocinas/genética , Citocinas/metabolismo , Duodeno/enzimologia , Perfilação da Expressão Gênica , Íleo/enzimologia , Íleo/metabolismo , Cadeias J de Imunoglobulina/genética , Cadeias J de Imunoglobulina/metabolismo , Cadeias alfa de Imunoglobulina/genética , Cadeias alfa de Imunoglobulina/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Muramidase/genética , Muramidase/metabolismo , Norepinefrina/sangue , Fosfolipases A2/genética , Fosfolipases A2/metabolismo , Receptores Fc/genética , Receptores Fc/metabolismo , Transcrição GênicaRESUMO
Caveolae are identifiable plasma membrane invaginations. The main structural proteins of caveolae are the caveolins. There are three caveolins expressed in mammals, designated Cav-1, Cav-2, and Cav-3. It has been postulated that Cav-1 acts as a scaffold protein for signaling proteins; these include ion channels, enzymes, and other ligand receptors like membrane-associated estrogen receptor (ER)alpha or ERbeta. Caveolae-associated membrane proteins are involved in regulating some of the rapid estrogenic effects of 17beta-estradiol. One important system related to the activity of ERalpha and caveolae is the renin-angiotensin system. Angiotensin II (ANG II) has numerous actions in vascular smooth muscle, including modulation of vasomotor tone, cell growth, apoptosis, phosphatidylinositol 3-kinase (PI3K)/Akt activation, and others. Many proteins associated with caveolae are in close relation with the scaffolding domain of Cav-1 (82-101 amino acid residues). It has been proposed that this peptide may acts as a kinase inhibitor. Therefore, to explore the ability of Cav-1 scaffolding peptide (CSP-1) to regulate ANG II function and analyze the relationship between ERalpha and ANG II type 1 and 2 (AT(1) and AT(2)) receptors, we decided to study the effects of CSP-1 on ANG II-induced intracellular Ca(2+) kinetics and the effect of 17beta-estradiol on this modulation using human smooth muscle cells in culture, intracellular Ca(2+) concentration measurements, immuno- and double-immunocytochemistry confocal analysis of receptor expression, immunoblot analysis, and immunocoprecipitation assays to demonstrate coexpression. We hypothesized that CSP-1 inhibits ANG II-mediated increases in intracellular Ca(2+) concentrations by interfering with intracellular signaling including the PI3K/Akt pathway. We also hypothesize that AT(2) receptors associate with Cav-1. Our results show that there is a close association of AT(1), AT(2), and ERalpha with Cav-1 in human arterial smooth muscle cells in culture. CSP-1 inhibits ANG II-induced intracellular signaling.
Assuntos
Angiotensina II/fisiologia , Cálcio/metabolismo , Caveolina 1/fisiologia , Estradiol/fisiologia , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , Caveolina 1/metabolismo , Células Cultivadas , Receptor alfa de Estrogênio/metabolismo , Humanos , Immunoblotting , Imuno-Histoquímica , Imunoprecipitação , Masculino , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor Tipo 1 de Angiotensina/metabolismo , Receptor Tipo 2 de Angiotensina/metabolismo , Transdução de Sinais/fisiologiaRESUMO
Autosomal recessive hypercholesterolemia (ARH) is characterized by elevated LDL serum levels, xanthomatosis, and premature coronary artery disease. Three loci have been described for this condition (1p35, 15q25-q26 and 13q). Recently, the responsible gene at the 1p35 locus, encoding an LDL receptor adaptor protein (ARH) has been identified. We studied a Mexican ARH family with two affected siblings. Sequence analysis of the ARH gene (1p35 locus) revealed that the affected siblings are homozygous for a novel mutation (IVS4+2T>G) affecting the donor splice site in intron 4, whereas both the parents and an unaffected sister are heterozygous for this mutation. The IVS4+2T>G mutation results in a major alternative transcript derived from a cryptic splice site, which carries an in-frame deletion of 78 nucleotides in the mature mRNA. The translation of this mRNA yields a mutant protein product (ARH-26) lacking 26 amino acids, resulting in the loss of beta-strands beta6 and beta7 from the PTB domain. This is the first case where a naturally occurring mutant with an altered PTB domain has been identified.