A nitroalkene derivative of salicylate alleviates diet-induced obesity by activating creatine metabolism and non-shivering thermogenesis.

Obesity-related type II diabetes (diabesity) has increased global morbidity and mortality dramatically. Previously, the ancient drug salicylate demonstrated promise for the treatment of type II diabetes, but its clinical use was precluded due to high dose requirements. In this study, we present a nitroalkene derivative of salicylate, 5-(2-nitroethenyl)salicylic acid (SANA), a molecule with unprecedented beneficial effects in diet-induced obesity (DIO). SANA reduces DIO, liver steatosis and insulin resistance at doses up to 40 times lower than salicylate. Mechanistically, SANA stimulated mitochondrial respiration and increased creatine-dependent energy expenditure in adipose tissue. Indeed, depletion of creatine resulted in the loss of SANA action. Moreover, we found that SANA binds to creatine kinases CKMT1/2, and downregulation CKMT1 interferes with the effect of SANA in vivo. Together, these data demonstrate that SANA is a first-in-class activator of creatine-dependent energy expenditure and thermogenesis in adipose tissue and emerges as a candidate for the treatment of diabesity.

Benefits in cardiac function by CD38 suppression: Improvement in NAD+ levels, exercise capacity, heart rate variability and protection against catecholamine-induced ventricular arrhythmias.

CD38 enzymatic activity regulates NAD+ and cADPR levels in mammalian tissues, and therefore has a prominent role in cellular metabolism and calcium homeostasis. Consequently, it is reasonable to hypothesize about its involvement in cardiovascular physiology as well as in heart related pathological conditions. AIM: To investigate the role of CD38 in cardiovascular performance, and its involvement in cardiac electrophysiology and calcium-handling. METHODS AND RESULTS: When submitted to a treadmill exhaustion test, a way of evaluating cardiovascular performance, adult male CD38KO mice showed better exercise capacity. This benefit was also obtained in genetically modified mice with catalytically inactive (CI) CD38 and in WT mice treated with antibody 68 (Ab68) which blocks CD38 activity. Hearts from these 3 groups (CD38KO, CD38CI and Ab68) showed increased NAD+ levels. When CD38KO mice were treated with FK866 which inhibits NAD+ biosynthesis, exercise capacity as well as NAD+ in heart tissue decreased to WT levels. Electrocardiograms of conscious unrestrained CD38KO and CD38CI mice showed lower basal heart rates and higher heart rate variability than WT mice. Although inactivation of CD38 in mice resulted in increased SERCA2a expression in the heart, the frequency of spontaneous calcium release from the sarcoplasmic reticulum under stressful conditions (high extracellular calcium concentration) was lower in CD38KO ventricular myocytes. When mice were challenged with caffeine-epinephrine, CD38KO mice had a lower incidence of bidirectional ventricular tachycardia when compared to WT ones. CONCLUSION: CD38 inhibition improves exercise performance by regulating NAD+ homeostasis. CD38 is involved in cardiovascular function since its genetic ablation decreases basal heart rate, increases heart rate variability and alters calcium handling in a way that protects mice from developing catecholamine induced ventricular arrhythmias.

SIRT6 stabilization and cytoplasmic localization in macrophages regulates acute and chronic inflammation in mice.

Acute and chronic inflammations are key homeostatic events in health and disease. Sirtuins (SIRTs), a family of NAD-dependent protein deacylases, play a pivotal role in the regulation of these inflammatory responses. Indeed, SIRTs have anti-inflammatory effects through a myriad of signaling cascades, including histone deacetylation and gene silencing, p65/RelA deacetylation and inactivation, and nucleotide­binding oligomerization domain, leucine rich repeat, and pyrin domain­containing protein 3 inflammasome inhibition. Nevertheless, recent findings show that SIRTs, specifically SIRT6, are also necessary for mounting an active inflammatory response in macrophages. SIRT6 has been shown to positively regulate tumor necrosis factor alpha (TNFα) secretion by demyristoylating pro-TNFα in the cytoplasm. However, how SIRT6, a nuclear chromatin-binding protein, fulfills this function in the cytoplasm is currently unknown. Herein, we show by Western blot and immunofluorescence that in macrophages and fibroblasts there is a subpopulation of SIRT6 that is highly unstable and quickly degraded via the proteasome. Upon lipopolysaccharide stimulation in Raw 264.7, bone marrow, and peritoneal macrophages, this population of SIRT6 is rapidly stabilized and localizes in the cytoplasm, specifically in the vicinity of the endoplasmic reticulum, promoting TNFα secretion. Furthermore, we also found that acute SIRT6 inhibition dampens TNFα secretion both in vitro and in vivo, decreasing lipopolysaccharide-induced septic shock. Finally, we tested SIRT6 relevance in systemic inflammation using an obesity-induced chronic inflammatory in vivo model, where TNFα plays a key role, and we show that short-term genetic deletion of SIRT6 in macrophages of obese mice ameliorated systemic inflammation and hyperglycemia, suggesting that SIRT6 plays an active role in inflammation-mediated glucose intolerance during obesity.

[Humoral immune response induced by influenza vaccine in children with acute lymphoblastic leukemia]. / Respuesta inmune humoral inducida por la vacuna influenza en niños con diagnóstico de leucemia linfoblástica aguda.

BACKGROUND: Patients with acute lymphoblastic leukemia (ALL) have high risk of severe influenza infection and vaccination is highly recommended. The immunogenicity and effectiveness of vaccination are lower than in healthy people. AIM: To evaluate the immune response induced by influenza vaccine in children with ALL and observe effectiveness. METHOD: Children with ALL in maintenance phase and healthy children were recruited. Blood samples were taken at vaccination day (D0) and at day 28 (D28). Humoral response was evaluated by hemaglutination inhibition test (HAI) against H1N1. Patients were followed up for one year, clinical data and influenza episodes were recorded. RESULTS: 34 children with ALL and 9 healthy children were included. Concerning HAI on D28, 12/34 patients and 5/8 healthy children had titers ≥ 1/40, with seroprotection rates of 35 and 63% respectively. Seroprotected children were older than non-seroprotected ones. During follow-up, only 3 patients non seroprotected, presented influenza infection, without oxygen supplementation or critical care support. DISCUSSION: Children with ALL had a lower seroprotection rate than healthy children. Nevertheless, none of the seroprotected children presented influenza infection, reinforcing the annual vaccination recommendation.

The protein Deleted in Breast Cancer-1 (DBC1) regulates vascular response and formation of aortic dissection during Angiotensin II infusion.

Cardiovascular diseases are among the main causes of morbimortality in the adult population. Among them, hypertension is a leading cause for stroke, heart disease and kidney failure. Also, as a result of arterial wall weakness, hypertension can lead to the development of dissecting aortic aneurysms, a rare but often fatal condition if not readily treated. In this work, we investigated the role of DBC1 in the regulation of vascular function in an ANGII-induced hypertension mouse model. We found that WT and DBC1 KO mice developed hypertension in response to ANGII infusion. However, DBC1 KO mice showed increased susceptibility to develop aortic dissections. The effect was accompanied by upregulation of vascular remodeling factors, including MMP9 and also VEGF. Consistent with this, we found decreased collagen deposition and elastic fiber fragmentation, suggesting that increased expression of MMPs in DBC1 KO mice weakens the arterial wall, promoting the formation of aortic dissections during treatment with ANGII. Finally, DBC1 KO mice had reduced cell proliferation in the intima-media layer in response to ANGII, paralleled with an impairment to increase wall thickness in response to hypertension. Furthermore, VSMC purified from DBC1 KO mice showed impaired capacity to leave quiescence, confirming the in vivo results. Altogether, our results show for the first time that DBC1 regulates vascular response and function during hypertension and protects against vascular injury. This work also brings novel insights into the molecular mechanisms of the development of aortic dissections.

Respuesta inmune humoral inducida por la vacuna influenza en niños con diagnóstico de leucemia linfoblástica aguda / Humoral immune response induced by influenza vaccine in children with acute lymphoblastic leukemia

Resumen Introducción: Los pacientes con leucemia linfoblástica aguda (LLA) tienen alto riesgo de influenza grave y la vacunación es altamente recomendada. La inmunogenicidad y efectividad de la vacuna es menor comparada a los sujetos sanos. Objetivo: Evaluar la respuesta inmune inducida por vacuna anti-influenza en niños con LLA y observar su efectividad. Métodos: Se reclutaron niños con LLA en terapia de mantención y niños sanos. Se tomaron muestras de sangre el día de la vacuna (D0) y al día 28 (D28), y se realizó test de inhibición de hemaglutinación (IHA) contra H1N1. Los pacientes fueron seguidos por un año, registrando datos clínicos y episodios de influenza. Resultados: Se incluyeron 34 niños con LLA y 9 niños sanos. Respecto al IHA en D28, 12/34 pacientes y 5/8 niños sanos presentaron títulos ≥ 1/40, resultando una tasa de seroprotección de 35 y 63%, respectivamente. Los niños seroprotegidos eran significativamente mayores. Durante el seguimiento, sólo tres pacientes, no seroprotegidos, presentaron infección por influenza, ninguno requirió oxigeno o cuidados intensivos. Discusión: Los niños con LLA alcanzaron una tasa seroprotección más baja que la observada en niños sanos. Sin embargo, ninguno de los niños seroprotegidos presentó infección por influenza, reforzando la recomendación de vacunación anual.

Abstract Background: Patients with acute lymphoblastic leukemia (ALL) have high risk of severe influenza infection and vaccination is highly recommended. The immunogenicity and effectiveness of vaccination are lower than in healthy people. Aim: To evaluate the immune response induced by influenza vaccine in children with ALL and observe effectiveness. Method: Children with ALL in maintenance phase and healthy children were recruited. Blood samples were taken at vaccination day (D0) and at day 28 (D28). Humoral response was evaluated by hemaglutination inhibition test (HAI) against H1N1. Patients were followed up for one year, clinical data and influenza episodes were recorded. Results: 34 children with ALL and 9 healthy children were included. Concerning HAI on D28, 12/34 patients and 5/8 healthy children had titers ≥ 1/40, with seroprotection rates of 35 and 63% respectively. Seroprotected children were older than non-seroprotected ones. During follow-up, only 3 patients non seroprotected, presented influenza infection, without oxygen supplementation or critical care support. Discussion: Children with ALL had a lower seroprotection rate than healthy children. Nevertheless, none of the seroprotected children presented influenza infection, reinforcing the annual vaccination recommendation.

A novel form of Deleted in breast cancer 1 (DBC1) lacking the N-terminal domain does not bind SIRT1 and is dynamically regulated in vivo.

The protein Deleted in Breast Cancer-1 is a regulator of several transcription factors and epigenetic regulators, including HDAC3, Rev-erb-alpha, PARP1 and SIRT1. It is well known that DBC1 regulates its targets, including SIRT1, by protein-protein interaction. However, little is known about how DBC1 biological activity is regulated. In this work, we show that in quiescent cells DBC1 is proteolytically cleaved, producing a protein (DN-DBC1) that misses the S1-like domain and no longer binds to SIRT1. DN-DBC1 is also found in vivo in mouse and human tissues. Interestingly, DN-DBC1 is cleared once quiescent cells re-enter to the cell cycle. Using a model of liver regeneration after partial hepatectomy, we found that DN-DBC1 is down-regulated in vivo during regeneration. In fact, WT mice show a decrease in SIRT1 activity during liver regeneration, coincidentally with DN-DBC1 downregulation and the appearance of full length DBC1. This effect on SIRT1 activity was not observed in DBC1 KO mice. Finally, we found that DBC1 KO mice have altered cell cycle progression and liver regeneration after partial hepatectomy, suggesting that DBC1/DN-DBC1 transitions play a role in normal cell cycle progression in vivo after cells leave quiescence. We propose that quiescent cells express DN-DBC1, which either replaces or coexist with the full-length protein, and that restoring of DBC1 is required for normal cell cycle progression in vitro and in vivo. Our results describe for the first time in vivo a naturally occurring form of DBC1, which does not bind SIRT1 and is dynamically regulated, thus contributing to redefine the knowledge about its function.

Right atrium cholinergic deficit in septic rats.

Heart rate variability (HRV) is mainly determined by the influence of both branches of the Autonomic Nervous System over the sinus node. Low HRV has been associated with a worse prognosis in patients with sepsis. The objective of this study was to explain the reduction in HRV during experimental sepsis in adult rats. We recorded the heart's electrical activity by telemetry in conscious unrestrained male rats before and 1day after the induction of peritonitis (N=39) or sham peritonitis (N=15). Then, we analyzed the chronotropic responsiveness of the isolated heart to the autonomic neurotransmitters and determined catecholamine concentrations in blood plasma and acetylcholine and choline concentrations in the right atrium. The surviving septic rats (N=33) had increased heart rate (HR) and diminished HRV. Despite the higher HR in situ, the spontaneous basal HR in septic and sham isolated hearts was the same. The isolated septic hearts showed acetylcholine hypersensitivity (log (IC50,M)=-7.2±0.2 vs. -6.0±0.4, P=0.025) and lower concentrations of choline in their right atriums (in nMol/mg protein: 0.6±0.1 vs. 1.6±0.6, P=0.013). Norepinephrine concentration in blood plasma from septic rats was higher (in ng/ml: 29.2±8.4 vs. 5.8±4.1, P=0.019). In conclusion, septic rats present a deregulation of the autonomic nervous system, not only sympathetic overexcitation but also parasympathetic dysfunction.

[Evaluation of quantitative cultures of endotracheal aspirates for pediatric ventilator-associated pneumonia diagnosis: analytic, prospective study]. / Uso de cultivo cuantitativo de aspirado endotraqueal para el diagnóstico de neumonía asociada a ventilación mecánica en pediatría: estudio prospectivo, analítico.

INTRODUCTION: Ventilator-associated pneumonia (VAP) is dificult to diagnose because of the absence of a gold standard. AIM: To evaluate the use of quantitative cultures of endotracheal aspirates for diagnosis of pediatric VAP and to obtain acceptable sensitivity and specificity cutoff points. PATIENTS AND METHODS: Prospective, analytic study which included patients under 15 years, who were connected to mechanical ventilation at Hospital Luis Calvo Mackenna's intensive care units. They were classified as cases or controls according to NNIS criteria. RESULTS: During a period of 21 months we recruited a total of 43 patients with a mean age of 16 months. We obtained endotracheal aspirated samples to perform quantitative cultures. Most frequently isolated pathogens were Staphylococcus aureus (34.5%) and non-fermentative Gram-negative bacilli (24.1%). We obtained a sensitivity of 88%, specificity of 70%, PPV of 82%, and NPV of 79% for samples greater than 10(6) CFU. CONCLUSIONS: Quantitative culture of endotracheal aspirates is a reliable method for diagnosing pediatric VAP when the value is greater than 10 CFU.

Asymmetrical properties of heart rate variability in type 1 diabetes.

Heart rate variability related to heart rate decelerations and accelerations is reduced in patients with type 1 diabetes compared to healthy individuals both in resting 10-min and ambulatory 24-h ECGs.

Correlations between frequency-domain HRV indices and lagged Poincaré plot width in healthy and diabetic subjects.

The conventional Poincaré plot for heart rate variability (HRV) analysis is a scatterplot of successive (lag 1) pairs of RR intervals (intervals between heartbeats), and its width (SD1) is considered a measure of short-term variability. It has been shown that SD1 correlates better with HF than with LF (high- and low-frequency bands of the spectrum, respectively). Our aim was to assess how these correlations were affected when SD1 was obtained for longer lags. 10 min ECGs were used to construct Poincaré plots with lags of 1-10 heartbeats in two groups of subjects, one with normal HRV and the other with impaired HRV (control and diabetic groups respectively, N = 15 each). SD1 was quantified for these subjects and HRV spectral indices were estimated. The diabetic group had lower LF, HF and SD1 than the control group (p < 0.05). In both groups, SD1 tended to increase as the lag increased. In the control group, SD1 for lags 1 and 2 was highly correlated with HF (r(s) > 0.9), while SD1 for lags 4 correlated better with LF (r(s) 0.9) than with HF (0.65

Nuclear factor kappa B activation in human cord blood mononuclear cells.

The immunologic signals participating in immune responses early in life have not been completely elucidated. Regarding the characterization of neonatal cells, little is known concerning the activity of transcription factor nuclear factor kappa B (NF-kappaB), which regulates inflammatory genes and cytokine production. The aim of this study was to characterize NF-kappaB activation in cord blood mononuclear cells (CBMC). We analyzed the potential association of NF-kappaB activity with lymphocyte proliferation and influences on cytokine secretion in the early immune system. To determine the contribution of a disease whereby inheritance may impact neonatal immunity, we assessed the influence of maternal allergic disease on NF-kappaB regulation and cytokine secretion. CBMC from healthy newborns were isolated and stimulated with mitogen (n = 28). Nuclear extracts were analyzed by electrophoretic mobility shift assay, cytokine secretion by ELISA. FISH analysis excluded relevant maternal contamination of CBMC. All samples showed a positive lymphoproliferative response, and NF-kappaB activity was both increased and decreased after mitogen stimulation. Increased NF-kappaB activation was significantly associated with decreased TNF-alpha secretion (median 6.1 versus 50.3 pg/mL) in unstimulated CBMC. Mitogen stimulation resulted in increased NF-kappaB activity with a trend to increased IL-13 production. Maternal allergic disease was associated with higher TNF-alpha (median 982 versus 173 pg/mL) and IL-13 secretion (median 1328 versus 1120 pg/mL) after mitogen stimulation. Together, NF-kappaB activity is differentially activated in cord blood and associated with a distinct cytokine pattern. Whether differential NF-kappaB activity in cord blood is related to the subsequent development of immune diseases requires further investigation.

Short-term studies of heart rate variability: comparison of two methods for recording.

Heart rate variability (HRV) is often analysed using short-term studies. Our objective was to compare two of them in a group of diabetic patients (reduced HRV) and in a control group. From the same 10 min surface electrocardiogram (ECG) two recordings were obtained. In one of them the whole signal was acquired through an A/D converter (post-event method). In the other (real-time method), an interface between the electrocardiograph and a parallel port of a computer was used to perform real-time processing of the ECG signal. The R-R intervals were measured after a visual validation in the post-event method. In the real-time method, the stored R-R intervals were automatically filtered. For both methods HRV indexes were calculated using the same software. The values of mean R-R intervals for each subject were almost identical regardless of the method. Accordingly, we found a high correlation between HRV indexes obtained from both methods (all Spearman values > or = 0.9441 and P < 0.0001). In addition, we found similar P values in the comparisons between the diabetic and control groups. We conclude that both methods are suitable for HRV analysis. Therefore, the selection of method can be based on other considerations such as the capability to store the ECG of the post-event method or the speed of analysis and lower cost of the real-time one.

Heart rate variability as early marker of multiple organ dysfunction syndrome in septic patients.

PURPOSE: To determine whether measuring heart rate variability (HRV) in a group of septic patients without multiple organ dysfunction syndrome (MODS) made it possible to predict which of them would later develop this syndrome. MATERIAL AND METHODS: We studied 46 septic patients without MODS at the time of admission to an intensive care unit (ICU). During the first 24 hours of admission, a 10-minute electrocardiogram (ECG) was performed and 8 HRV indexes were calculated off-line. Eleven patients later developed MODS (MODS group) during their ICU stay, and 28 did not (non-MODS group). Seven patients were excluded. RESULTS: Although Acute Physiological and Chronic Health Evaluation (APACHE II) scores were similar for both groups, most HRV indices on admission were reduced significantly in the MODS group. Compared with a subset from the non-MODS group (control group, n = 11) paired by age, the MODS group had significantly lower low-frequency spectral components (LF, P =.0128) and mean squared successive differences of R-R intervals (rMSSD) (P =.0473) values. Multivariable logistic regression identified LF as the best predictor of MODS and received operating characteristic (ROC) curves established its cut-off point at 18 ms(2). Mortality rates were 63.6% for the MODS group and 0% for the non-MODS group (P <.0001). CONCLUSIONS: Reduction of HRV on ICU admission may be useful in identifying septic patients at risk for development of MODS.

Heart rate variability: short-term studies are as useful as holter to differentiate diabetic patients from healthy subjects.

BACKGROUND: The definitive incorporation of heart rate variability (HRV) as a clinical tool depends on the development of more confident techniques of measurement. The length of the studies is a critical issue. Whereas Holter studies allow the monitorization at different hours and activities, short-term recordings allow the control of environmental conditions. Recording length is also strongly related to the procedure of analysis; for instance, some time-domain indexes are strongly affected by the duration of the study. Meanwhile, spectral analyses require stationary conditions, only achieved in short-term studies. Our main goal was to determine if HRV indexes obtained from short-term analyses were as useful as those from Holter monitoring for diagnosis of reduced HRV in diabetes. METHODS: We studied two groups: one with impaired HRV (15 diabetic patients) and another with normal HRV (15 healthy subjects). HRV indexes obtained from 24-hour Holter recordings (SDNN, rMSSD, and the power of LF and HF bands), were correlated with analog indexes obtained from 10-minute digital acquired studies within each group. Besides, we compared the diabetic and control groups using the indexes obtained with both methodologies. RESULTS: The correlation was high (0.70

Evaluación del serodiagnóstico en el absceso hepático amebiano / Serodiagnostic evaluation for amebic liver abscess

El presente estudio fue diseñado para evaluar la eficacia del diagnóstico serológico del absceso hepático amibiano (AHA). Se utilizó una prueba de Elisa para la detección en suero de IgG específica, utilizando una fase sólida con una alta capacidad de adherencia. Se estudiaron 147 personas; 22 pacientes con sospecha clínica y ultrasonográfica de AH, 30 individuos completamente sanos, 9 portadores asintomáticos de E. histolytica, 35 con colitis amibiana pasada, 35 con otras parasitosis intestinales, 9 con otras patologías hepáticas y 6 con colitis amebiana presente. El rendimiento global de la prueba fue analizado por medio de las curvas del receptor-operador y del área bajo la curva. Se determinó que existe una diferencia significativa en la densidad óptica (DO) de los siete grupos estudiados (Kruskal-Wallis entre todos los grupos: p=0,0001); esta diferencia no existe cuando, al comparar, se elimina el grupo 1 del análisis (Kruskal-Wallis exceptuando el grupo 1: p=0,8203). El área bajo la curva ROC fue igual a 0,9941, dato muy cercano al ideal que es 1. Se recomienda la utilización de esta fase sólida teniendo en cuenta la importancia de la fase de bloqueo con albúmina de huevo. Se determinó que, a pesar de estar en una zona donde el diagnóstico de amebiasis es frecuente, las infecciones intestinales por esta ameba no presentan niveles detectables de IgG específica contra E. histolytica por esta prueba. Sin embargo, es importante ampliar la cantidad de personas estudiadas en estos grupos, principalmente, en el grupo de pacientes con otras patologías hepáticas puesto que este grupo incluye las entidades clínicas con las que realmente se debe hacer diagnóstico diferencial de (AHA) (absceso hepático piógeno (AHP), principalmente) para poder realizar la última fase de la evaluación. El estudio de este grupo nos ayudaría a obtener valores reales de sensibilidad, especificidad y valores predictivos y nos darían una estimación real de la capacidad discriminatoria de la prueba para obtener el verdadero valor del área bajo la curva y no el casi ideal que informamos aquí