Candida Variety in the Oral Cavity of Mexican Subjects with Type 2 Diabetes Mellitus and TLR2 Gene Expression.

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.

Recognition of the interaction between the bioactive peptide Val-Pro-Pro and the minimal promoter region of genes SOD and CAT using QCM-D and docking studies.

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.

Anti-Inflammatory Effect of Ethanolic Extract from Tabebuia rosea (Bertol.) DC., Quercetin, and Anti-Obesity Drugs in Adipose Tissue in Wistar Rats with Diet-Induced Obesity.

Obesity is characterized by the excessive accumulation of fat, which triggers a low-grade chronic inflammatory process. Currently, the search for compounds with anti-obesogenic effects that help reduce body weight, as well as associated comorbidities, continues. Among this group of compounds are plant extracts and flavonoids with a great diversity of action mechanisms associated with their beneficial effects, such as anti-inflammatory effects and/or as signaling molecules. In the bark of Tabebuia rosea tree, there are different classes of metabolites with anti-inflammatory properties, such as quercetin. Therefore, the present work studied the effect of the ethanolic extract of T. rosea and quercetin on the mRNA of inflammation markers in obesity compared to the drugs currently used. Total RNA was extracted from epididymal adipose tissue of high-fat diet-induced obese Wistar rats treated with orlistat, phentermine, T. rosea extract, and quercetin. The rats treated with T. rosea and quercetin showed 36 and 31% reductions in body weight compared to the obese control, and they likewise inhibited pro-inflammatory molecules: Il6, Il1b, Il18, Lep, Hif1a, and Nfkb1 without modifying the expression of Socs1 and Socs3. Additionally, only T. rosea overexpressed Lipe. Both T. rosea and quercetin led to a reduction in the expression of pro-inflammatory genes, modifying signaling pathways, which led to the regulation of the obesity-inflammation state.

Catecholamine levels and gene expression of their receptors in tissues of adults with osteosarcoma.

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.

Modulation of the mRNA of the Nlrp3 inflammasome by Morin and PUFAs in an obesity model induced by a high-fat diet.

Morin and PUFAs are bioactive compounds provided by the diet, with multiple biological activities, among which are the modulation of inflammation in various chronic diseases. The effect of supplementation with Morin, PUFAs, and the mixture of both on the levels of mRNA expression of the Nlrp3 inflammasome as well as genes associated with inflammation and lipid metabolism, in an obesity model through a high-fat diet, during 8 weeks of administration were evaluated. The three treatments negatively regulated the expression of Nlrp3 mRNA. Morin showed a better effect by modulating downwards the expression of the mRNA of Il-18, Casp-1, Pparγ, and Serbp-1c, in addition to positively modulating the expression of the mRNA of Ppar-α, as well as Adiponectin. The combined treatment of Morin plus the PUFAs maintained similar levels under normal conditions for the mRNA expression of Tlr4 and Ucp2.

DNA methylation of the RE-1 silencing transcription factor in peripheral blood mononuclear cells and gene expression of antioxidant enzyme in patients with late-onset Alzheimer disease.

Late-onset Alzheimer disease (LOAD) is the most frequent cause of dementia in elderly adults. However, the factors determining disease onset remain unclear. In the elderly, the activation and expression of the gene encoding RE-1 silencing transcription factor (REST) may be a determinant of neuroprotective mechanisms and good amyloidogenic pathway management. In the present study, the minimal promoter region of REST1 was genetically and epigenetically analyzed in blood samples from 21 subjects with LOAD and 20 cognitively healthy elderly subjects. Genomic DNA was isolated, treated with bisulfite and pyrosequenced, and gene expression was determined using real-time PCR. Notably, subjects with LOAD exhibited hypermethylation and significantly diminished expression of REST1 compared with healthy subjects (p = 0.001). In the LOAD group, the gene expression of CAT, SOD2 and GPX also showed a significant decrease and an increase in malondialdehyde. A docking analysis revealed that the first zinc finger protein Sp1 recognized and bound the methylated sequence in subjects with LOAD differently than the binding observed in control subjects. These results reveal that in patients with LOAD the methylation of specific sites in the promoter sequence of REST suppresses its expression and this could be regulating the decreased expression of CAT, SOD and GPX, besides interfering with the action of transcription factors as Sp1.

Intense craving for appetizing foods: validation and standardization of the Food Cravings Questionnaire-Trait in Mexico.

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.

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.

Deseo intenso por alimentos apetecibles: validación y estandarización del Food Craving Questionnaire-Trait en México / Intense craving for appetizing foods: validation and standardization of the Food Cravings Questionnaire-Trait in Mexico

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.

Neuroprotective Effects of Apocynin and Galantamine During the Chronic Administration of Scopolamine in an Alzheimer's Disease Model.

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.

Design, synthesis, molecular docking and in vitro evaluation of benzothiazole derivatives as 11ß-hydroxysteroid dehydrogenase type 1 inhibitors.

11-Beta hydroxysteroid dehydrogenase type 1 (11ß-HSD1) regulates cortisol levels mainly in adipose, hepatic and brain tissues. There is a relationship between the high activity of this enzyme and the development of obesity and metabolic disorders. The inhibition of 11ß-HSD1 has been shown to attenuate the development of type 2 diabetes mellitus, insulin resistance, metabolic syndrome and other diseases mediated by excessive cortisol production. In this work, fifteen benzothiazole derivatives substituted with electron-withdrawing and electron-donating groups were designed to explore their affinity for 11ß-HSD1 using in silico methods. The results show that (E)-5-((benzo[d]thiazol-2-ylimino)(methylthio)methylamino)-2-hydroxybenzoic acid (C1) has good physicochemical properties and favorable interactions with 11ß-HSD1 through hydrogen bonding and hydrophobic interactions in the catalytic site formed by Y183, S170 and Y177. Furthermore, C1 was synthesized and evaluated in vitro and ex vivo using clobenzorex (CLX) as a reference drug in obese Zucker rats. The in vitro results showed that C1 was a better inhibitor of human 11ß-HSD1 than CLX. The ex vivo assay results demonstrated that C1 was capable of reducing 11ß-HSD1 overexpression in mesenteric adipose tissue. Therefore, C1 was able to decrease the activity and expression of 11ß-HSD1 better than CLX.

Biosensor of Inflammation Biomarkers Based on Electrical Bioimpedance Analysis on Immobilized DNA Without Chemical Modification.

The development of biosensors to identify molecular markers or specific genes is fundamental for the implementation of new techniques that allow the detection of specific Deoxyribonucleic acid (DNA) sequences in a fast, economic and simple way. Different detection techniques have been proposed in the development of biosensors. Electrical Bioimpedance Spectroscopy (EBiS) has been used for diagnosis and monitoring of human pathologies, and is recognized as a safe, fast, reusable, easy and inexpensive technique. This study proves the development of a complementary DNA (cDNA) biosensor based on measurements of EBiS and DNA's immobilization with no chemical modifications. The evaluation of its potential utility in the detection of the gene expression of three inflammation characteristic biomarkers (NLRP3, IL-1ß and Caspase 1) is presented. The obtained results demonstrate that EBiS can be used to identify different gene expression patterns, measurements that were validated by Quantitative Polymerase Chain Reaction (qPCR). These results indicate the technical feasibility for a biosensor of specific genes through bioimpedance measurements on the immobilization of cDNA.

Early Detection of Vascular Obstruction in Microvascular Flaps Using a Thermographic Camera.

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.

Virome and bacteriome characterization of children with pneumonia and asthma in Mexico City during winter seasons 2014 and 2015.

BACKGROUND: Acute asthma exacerbations and pneumonia are important causes of morbidity and mortality in children and may coexist in the same children, although symptom overlap may lead to difficulties in diagnosis. Microbial and viral diversity and differential abundance of either may play an important role in infection susceptibility and the development of acute and chronic respiratory diseases. OBJECTIVES: To describe the virome and bacteriome present in the upper respiratory tract of hospitalized children with a clinical diagnosis of asthma and pneumonia during an acute exacerbation and an acute respiratory illness ARI episode respectively. METHODS: During the winter seasons of 2013-2014 and 2014-2015, 134 nasopharyngeal swabs samples of children <15 years of age with ARI hospitalized at a referral hospital for respiratory diseases were selected based on clinical diagnosis of asthma or pneumonia. The virome and bacteriome were characterized using Whole Genome Sequencing (WGS) and in-house bioinformatics analysis pipeline. RESULTS: The Asthma group was represented mainly by RV-C, BoV-1 and RSV-B and the pneumonia group by Bacteriophage EJ-1 and TTMV. TTV was found in both groups with a similar amount of reads. About bacterial composition Moraxella catarrhalis, Propionibacterium acnes and Acinetobacter were present in asthma and Veillonella parvula and Mycoplasma pneumoniae in pneumonia. Streptococcus pneumoniae and Haemophilus influenzae were mostly found with both asthma and pneumonia. CONCLUSIONS: Our results show a complex viral and bacterial composition in asthma and pneumonia groups with a strong association of RV-C presence in asthmatic children. We observed Streptococcus pneumoniae and Haemophilus influenzae concurrently in both groups.

Different behavior of myeloperoxidase in two rodent amoebic liver abscess models.

The protozoan Entamoeba histolytica is the etiological agent of amoebiasis, which can spread to the liver and form amoebic liver abscesses. Histological studies conducted with resistant and susceptible models of amoebic liver abscesses (ALAs) have established that neutrophils are the first cells to contact invasive amoebae at the lesion site. Myeloperoxidase is the most abundant enzyme secreted by neutrophils. It uses hydrogen peroxide secreted by the same cells to oxidize chloride ions and produce hypochlorous acid, which is the most efficient microbicidal system of neutrophils. In a previous report, our group demonstrated that myeloperoxidase presents amoebicidal activity in vitro. The aim of the current contribution was to analyze in vivo the role of myeloperoxidase in a susceptible (hamsters) and resistant (Balb/c mice) animal models of ALAs. In liver samples of hamsters and mice inoculated intraportally with Entamoeba histolytica trophozoites, the number of neutrophils in ALAs was determined by enzymatic activity. The presence of myeloperoxidase was observed by staining, and its expression and activity were quantified in situ. A significant difference existed between the two animal models in the number of neutrophils and the expression and activity of myeloperoxidase, which may explain the distinct evolution of amoebic liver abscesses. Hamsters and mice were treated with an MPO inhibitor (4-aminobenzoic acid hydrazide). Hamsters treated with ABAH showed no significant differences in the percentage of lesions or in the percentage of amoebae damaged compared with the untreated hamsters. ABAH treated mice versus untreated mice showed larger abscesses and a decreased percentage of damaged amoebae in these lesion at all stages of evolution. Further studies are needed to elucidate the host and amoebic mechanisms involved in the adequate or inadequate activation and modulation of myeloperoxidase.

Cytokine profile of NALT during acute stress and its possible effect on IgA secretion.

Stress stimuli affect the immune system responses that occur at mucosal membranes, particularly IgA secretion. It has been suggested that acute stress increases the levels of IgA and that sympathetic innervation plays an important role in this process. We herein explore in a murine model how acute stress affects the Th1/Th2/Treg cytokine balance in NALT, and the possible role of glucocorticoids in this effect. Nine-week-old male CD1 mice were divided into three groups: unstressed (control), stressed (subjected to 4h of immobilization), and stressed after pretreatment with a single dose of the corticosterone receptor antagonist RU-486. The parameters evaluated included plasma corticosterone and epinephrine, IgA levels in nasal fluid (by ELISA), the percentage of CD19+B220+IgA+ lymphocytes and CD138+IgA+ plasma cells, and the mRNA expression of heavy α chain, J chain and pIgR. Moreover, the gene and protein expression of Th1 cytokines (TNFα, IL-2 and INF-γ), Th2 cytokines (IL-4 and IL-5) and Treg cytokines (IL-10 and TGFß) were determined in nasal mucosa. The results show that acute stress generated a shift towards the dominance of an anti-inflammatory immune response (Th2 and Treg cytokines), evidenced by a significant rise in the amount of T cells that produce IL4, IL-5 and IL-10. This immune environment may favor IgA biosynthesis by CD138+IgA+ plasma cells, a process mediated mostly by glucocorticoids.

Undaria pinnatifida and Fucoxanthin Ameliorate Lipogenesis and Markers of Both Inflammation and Cardiovascular Dysfunction in an Animal Model of Diet-Induced Obesity.

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.

n-3 Fatty acids modulate the mRNA expression of the Nlrp3 inflammasome and Mtor in the liver of rats fed with high-fat or high-fat/fructose diets.

CONTEXT: There is evidence that n-3 polyunsaturated fatty acids (n-3-PUFAs) can inhibit mTORC1, which should potentiate autophagy and eliminate NLRP3 inflammasome activity. OBJECTIVE: Evaluate the effect of a high-fat or high-fat/fructose diet with and without n-3-PUFAs on hepatic gene expression. MATERIALS AND METHODS: We examined the mRNA expression by RT-PCR of Mtor, Nlrp3, and other 22 genes associated with inflammation in rats livers after a 9-week diet. The dietary regimens were low-fat (control, CD), high-fat (HF), high-fat/fructose (HF-Fr), and also each of these supplemented with n-3-PUFAs (CD-n-3-PUFAs, HF-n-3-PUFAs, and HF-Fr-n-3-PUFAs). These data were processed by GeneMania and STRING databases. RESULTS: Compared to the control, the HF group showed a significant increase (between p < 0.05 and p < 0.0001) in 20 of these genes (Il1b, Il18, Rxra, Nlrp3, Casp1, Il33, Tnf, Acaca, Mtor, Eif2s1, Eif2ak4, Nfkb1, Srebf1, Hif1a, Ppara, Ppard, Pparg, Mlxipl, Fasn y Scd1), and a decrease in Sirt1 (p < 0.05). With the HF-Fr diet, a significant increase (between p < 0.05 and p < 0.005) was also found in the expression of 16 evaluated genes (Srebf1, Mlxipl, Rxra, Abca1, Il33, Nfkb1, Hif1a, Pparg, Casp1, Il1b, Il-18, Tnf, Ppard, Acaca, Fasn, Scd1), along with a decrease in the transcription of Mtor and Elovl6 (p < 0.05). Contrarily, many of the genes whose expression increased with the HF and HF-Fr diets did not significantly increase with the HF-n-3-PUFAs or HF-Fr-n-3-PUFAs diet. DISCUSSION AND CONCLUSION: We found the interrelation of the genes for the mTORC1 complex, the NLRP3 inflammasome, and other metabolically important proteins, and that these genes respond to n-3-PUFAs.

Oxidative damage and antioxidant defense in thymus of malnourished lactating rats.

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.

17ß-Estradiol transcriptionally modulates Nlrp1 and Nlrp3 inflammasomes in gonadectomized rats with inflammation.

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.

[Carotid and brachial intima-media thickness of the allelic variants G894T and T-789C of eNOS in systemic sclerosis patients]. / Grosor íntima-media de las variantes alélicas G894T y T-789C de eNOS en pacientes con esclerosis sistémica.

OBJECTIVE: To evaluate comparatively the carotid intima media thickness index (IMT) and brachial index with Doppler ultrasound, and pulmonary artery pressure with echocardiography in allelic variation of G894T and T-789C eNOS (endothelial nitric oxide synthase) in patients with systemic sclerosis. METHODS: This is a cross-sectional study in patients with scleroderma. The inclusion criteria were: age 18-70 years, scleroderma diagnosed patients with a prior informed consent. Polymorphism G894T and T-789C of eNOS was measured in blood, and IMT by carotid and brachial Doppler. We analyzed with descriptive statistics, Student's t test or chi square for association of variables and Spearman to correlate Doppler parameters. RESULTS: We found abnormally high IMT in carotid and brachial arteries. We also observed low correlation between both brachial arteries, good correlation between carotids and no correlation between carotid and brachial arteries. The left IMT>0.65 mm in brachial artery corresponds to pulmonary pressure>30 mmHg by echocardiography. G894T gene polymorphism was associated with increased IMT in right carotid. CONCLUSION: The G894T eNOS polymorphism was associated with increased IMT in right carotid. The one side carotid IMT is consistent with its contralateral. The left brachial IMT above to 0.65 mm, suggests the possibility of pulmonary arterial hypertension.

Introducción: el objetivo de este estudio es evaluar comparativamente el grosor íntima-media (IMT) carotídeo y braquial con ultrasonido Doppler, la presión arterial pulmonar ecocardiográfica y la variación alélica de los genes G894T y T-789C de la eNOS en pacientes con esclerodermia. Métodos: estudio transversal en pacientes con esclerodermia. Criterios de inclusión: edad 18-70 años, diagnosticado con esclerodermia, previo consentimiento informado. Se evaluó IMT, índice de resistencia (IR) e índice de pulsatilidad (IP) mediante Doppler carotídeo y braquial, perfil de lípidos, proteína C reactiva, polimorfismo G894T y T-789C de óxido nítrico sin tasa endotelial (eNOS). Analizamos con estadística descriptiva, prueba t de Student o chi cuadrada para asociación de variables y Spearman para correlacionar las variables cuantitativas. Resultados: encontramos IMT anormalmente alto en arterias carotídeas y braquiales. Observamos baja correlación entre ambas braquiales, buena correlación entre carótidas y ninguna correlación entre carótidas y braquiales. El IMT braquial izquierdo > 0.65 corresponde con presión arteria pulmonar > 30 mmHg ecocardiográfica. El polimorfismo del gen G894T se asoció con mayor IMT en carótida derecha.Conclusión: El polimorfismo G894T de eNOS se asocia a mayor IMT en carótida derecha. El IMT carotídeo concuerda con su IMT contralateral. El IMT braquial izquierdo mayor a 0.65 sugiere la posibilidad de hipertensión arterial pulmonar.