Hyperglycemia affects neuronal differentiation and Nestin, FOXO1, and LMO3 mRNA expression of human Wharton's jelly mesenchymal stem cells of children from diabetic mothers.

Pregestational Diabetes Mellitus (PDM) during pregnancy constitutes an unfavorable embryonic and fetal development environment, with a high incidence of congenital malformations (CM). Neural tube defects are the second most common type of CM in children of diabetic mothers (CDM), who also have an elevated risk of developing neurodevelopmental disorders. The mechanisms that lead to these neuronal disorders in CDM are not yet fully understood. The present study aimed to know the effect of hyperglycemia on proliferation, neuronal differentiation percentage, and expression of neuronal differentiation mRNA markers in human umbilical cord Wharton's jelly mesenchymal stem cells (hUCWJMSC) of children from normoglycemic pregnancies (NGP) and PDM. We isolated and characterized hUCWJMSC by flow cytometry, immunofluorescence, RT-PCR and were induced to differentiate into adipocytes, osteocytes, and neurons. Proliferation assays were performed to determine the doubling time, and Nestin, TUBB3, FOXO1, KCNK2, LMO3, and MAP2 mRNA gene expression was assessed by semiquantitative RT-PCR. Hyperglycemia significantly decreased proliferation and neuronal differentiation percentage in NGP and PDM cells treated with 40 mM d-glucose. Nestin mRNA expression decreased under control glycemic conditions, while FOXO1, KCNK2, LMO3, and MAP2 mRNA expression increased during neuronal differentiation in both NGP and PDM cells. On the other hand, under hyperglycemic conditions, Nestin was significantly decreased in cells from NGP but not in cells from PDM, while mRNA expression of FOXO1 and LMO3 was significantly increased in cells from NGP, but not in cells from PDM. We found evidence that maternal PDM, with hyperglycemia in culture, affects the biological properties of fetal cells. All these results could be part of fetal programming.

DASH Diet as a Proposal for Improvement in Cellular Immunity and Its Association with Metabolic Parameters in Persons with Overweight and Obesity.

The development of obesity entails a chronic low-grade inflammatory state with increased pro-inflammatory cells, mainly in visceral adipose tissue (VAT). Additionally, dietary patterns have an influence on the regulation of chronic inflammation. Dietary Approaches to Stop Hypertension (DASH) include foods with an anti-inflammatory profile and that have positive impacts on body composition (BC), suggesting improvements in inflammatory processes. OBJECTIVE: To analyze the impact of the DASH diet on cellular immunity, anthropometric, biochemical and BC parameters in patients with overweight and obesity, who could present metabolic syndrome. METHODOLOGY: Lymphocyte subpopulations, biochemical parameters, anthropometric parameters, and BC before and 8 weeks after intervention with the DASH diet in persons with overweight and obesity were measured. RESULTS: Fifty-nine young adults participated in the study. After the intervention, no significant changes in biochemical parameters were observed, although a significant decrease in nearly all of the anthropometric and BC variables was found: waist circumference (p < 0.001), percentage and kilograms of fat (p < 0.001 and p < 0.025, respectively), VAT (p < 0.020), and weight (p < 0.001), as well as total lymphocytes and double-positive TCD4+ cells. A relation between changes in leukocyte subpopulations (monocytes, natural killer, helper and cytotoxic lymphocytes, and naive TCD4+ cells) and metabolic improvements (glucose, triglycerides, total cholesterol and LDL-c) was also found. CONCLUSIONS: The DASH diet promotes positive changes in lymphocyte subpopulations, anthropometric parameters and BC in persons with overweight and obesity. Future studies should elucidate the cellular and molecular mechanisms through which the DASH diet produces inmunometabolic improvement.

t-BHQ Protects Against Oxidative Damage and Maintains the Antioxidant Response in Malnourished Rats.

OBJECTIVE: Tert-butylhydroquinone (t-BHQ) protective effect against oxidative damage in thymus from malnourished pops-rats was evaluated. METHODS: Malnutrition in pops-rats was induced during the lactation period and first-, second-, and third-degree malnourished rats were studied (MN1, MN2, and MN3). To determine t-BHQ protective effect, lipid peroxidation (LPx) was assessed, as well as the carbonyl content. The reduced glutathione and glutathione disulfide content were determined and antioxidant enzyme activities were measured. RESULTS: Oxidative protein damage, LPx, and Nuclear Factor-κB (NF-κB) content, increased in the MN2 and MN3 compared to well-nourished rats, associated with lower protein content and antioxidant activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase. Tert-butylhydroquinone treatment induced a protective effect against lipids and proteins oxidative damage, as well as decrease in NF-κB in MN rats and restored the antioxidant mechanisms, mostly GPx and SOD. No differences were found between male and female animals. CONCLUSIONS: Results show that higher body weight deficit leads to increased oxidative damage and probably inflammation, attributable to alterations in antioxidant mechanisms. These effects were reversed by the t-BHQ-treatment, which restores the antioxidant response. Our findings suggest that t-BHQ could be an interesting pharmacological intervention, but it needs to be studied further.

Mecanismos inmunológicos involucrados en la obesidad / Immune mechanisms involved in obesity

La obesidad es un problema de salud pública creciente a nivel mundial. Se ha clasificado como una enfermedad ocasionada por acumulación excesiva de triglicéridos en el tejido adiposo (TA). El TA visceral (TAV), se considera un factor importante en el desarrollo de enfermedades crónicas no transmisibles. Esto principalmente porque el adipocito aumenta en tamaño y número, lo cual lleva a hipoxia, liberación de ácidos grasos, movilización y activación de subpoblaciones leucocitarias (linfocitos T, B, macrófagos, neutrófilos y eosinófilos), liberación de mediadores proinflamatorios (factor de necrosis tumoral- α (TNF-α), interleucina (IL)-6, resistina) y disminución en la secreción de antiinflamatorios (adiponectina, IL-10, IL-4 e IL-13). Estos cambios en el TAV generan un estado de inflamación crónica de baja intensidad, que se ha relacionado con resistencia a la insulina (RI). La RI es el signo fundamental para el desarrollo del Síndrome Metabólico (SM), que inicialmente se presenta localmente en el TAV y después se vuelve sistémica. En la investigación de la obesidad y el desarrollo de sus comorbilidades, la trascendencia del TAV en la interacción entre células adiposas e inmunitarias, así como de liberación de mediadores para desencadenar el proceso inflamatorio crónico, es clave para el desarrollo de RI y SM; sin embargo, se ha observado que la masa músculo-esquelética está implicada en este proceso, aparte de las células y mediadores de los que se sabe su participación. De esta manera a los linfocitos T CD4 y T CD8 se les señala como proinflamatorios, en cambio a los eosinófilos y a la adiponectina se les clasifica como protectores del proceso.

Obesity is a widespread public health problem worldwide. This disease has been classified as a condition caused by excessive accumulation of triglycerides in adipose tissue (TA). Visceral TA is an important factor in the development of several chronic diseases, since the adipocyte increases in size and number, leading to hypoxia, fatty acid release, mo-bilization and activation of leukocyte subpopulations (macrophages, neutrophils, eosinophils, NK cells, T lymphocytes and B); release of pro-inflammatory mediators (TNF-á, IL-6, PAI-1, resistin and visfatin), and decreased secretion of anti-inflammatory cytokines (adiponectin, IL-10, IL-4 and IL-13). These changes in TAV generate a chronic inflammation state of low inten-sity, that has been linked to insulin resistance (IR), initially local and then becomes systemic, which represents the determining factor for the development of metabolic syndrome (SM). The transcendence of TAV in the interaction between adipose and immune cells, as well as the release of mediators that trigger the chronic inflammatory process, is key for the development of IR and SM. However, in the investigation of obesity and the development of its comorbidities, it has been observed that the skeletal-muscle mass is involved in this process, apart from the cells and mediators of which their participation is known. In this way, TCD4 and TCD8 lymphocytes are recognized as pro-inflammatory. In contrast, eosinophils and adiponectin are considered as protective of the process.

Infected malnourished children displayed changes in early activation and lymphocyte subpopulations.

AIM: Malnutrition and infections cause immunological changes in lymphocyte subpopulations and their functionality. We evaluated the activation capacity of lymphocytes and memory cells in 10 well nourished, seven well-nourished infected and eight malnourished infected children before and after treatment. METHODS: All the children were patients in Mexico City and were less than three years of age. The expression of various cluster of differentiation (CD) cells was assessed by flow cytometry: CD45RA (naïve) and CD45RO (memory) antigens on CD4 lymphocytes and CD69 in all lymphocytes. RESULTS: Well-nourished infected children showed a higher percentage of activated T lymphocyte (T cells), CD8+ and CD4+ memory cells during the infectious phase, suggesting that the activation mechanisms were triggered by infection. T cells from malnourished infected children showed a lower percentage of activated and memory cells. The T cell population size returned to baseline during the resolution phase of the infection in well-nourished infected children, but their T, B lymphocyte and natural killer (NK) cell counts remained high. In malnourished infected children, activated NK cells counts were low before and after therapy. CONCLUSION: After therapy, malnourished infected children showed poor NK cell responses during the infection's resolution phase, suggesting a persistent malnutrition-mediated immunological deficiency.

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.