Divergent effects of the antiretroviral drugs, dolutegravir, tenofovir alafenamide, and tenofovir disoproxil fumarate, on human adipocyte function.

Combined antiretroviral therapy (cART) has been associated with increased body weight accompanied by metabolic alterations in people living with human immunodeficiency virus (PLWH). To gain insight into the combined effects of cART components on adipocyte dysfunction, we assessed whether and how treatment of human adipocytes with dolutegravir (DTG) and the nucleotide-analog reverse-transcriptase inhibitors (NRTIs), tenofovir alafenamide (TAF) and tenofovir disoproxil fumarate (TDF), alone and in combination, altered biological processes related to adipose tissue dysfunction. DTG, TAF, and TDF were applied to human Simpson-Golabi-Behmel syndrome (SGBS) adipose cells during differentiation (day 10) and ensuing differentiation (day 14). Expression of selected marker genes was determined by qPCR, the release of adipokines and inflammatory cytokines to the culture media was assessed, and cell respiration was measured. Adipogenesis was not altered by the combined treatment of human adipocytes. However, DTG at the highest dose repressed adipogenesis marker genes expression, and TAF and TDF appeared to mitigate this effect. DTG repressed the expression of adiponectin and the release of adiponectin and leptin in differentiating adipocytes, and these effects were mantained in combination with TAF and TDF. DTG plus TAF or TDF on human adipocytes enhanced inflammation and stress and increased the release of proinflammatory cytokines to the culture media. Together, our results show that combined therapy with these drugs can alter inflammation, cellular stress, and fibrosis in human adipocytes. These findings may improve our understanding and management of the effects of cART on body adiposity and metabolic dysregulation in PLWH.

Inflammation of brown/beige adipose tissues in obesity and metabolic disease.

Many of the comorbidities of obesity, including type 2 diabetes and cardiovascular diseases, are related to the low-grade chronic inflammation of white adipose tissue. Under white adipocyte stress, local infiltration of immune cells and enhanced production of pro-inflammatory cytokines together reduce metabolic flexibility and lead to insulin resistance in obesity. Whereas white adipocytes act in energy storage, brown and beige adipocytes specialize in energy expenditure. Brown and beige activity protects against obesity and associated metabolic disorders, such as hyperglycaemia and hyperlipidaemia. Compared to white fat, brown adipose tissue depots are less susceptible to developing local inflammation in response to obesity; however, strong obesogenic insults ultimately induce a locally pro-inflammatory environment in brown fat. This condition directly alters the thermogenic activity of brown fat by impairing its energy expenditure mechanism and uptake of glucose for use as a fuel substrate. Pro-inflammatory cytokines also impair beige adipogenesis, which occurs mainly in subcutaneous adipose tissue. There is evidence that inflammatory processes occurring in perivascular adipose tissues alter their brown-versus-white plasticity, impair the extent of browning in these depots and favour the local release of vasculature damaging signals. In summary, the targeting of brown and beige adipose tissues by pro-inflammatory signals and the subsequent impairment of their thermogenic and metabolite draining activities appears to represent obesity-driven disturbances that contribute to metabolic syndrome and cardiovascular alterations in obesity.

Autophagic control of cardiac steatosis through FGF21 in obesity-associated cardiomyopathy.

OBJECTIVE: High-fat diet-induced obesity leads to the development of hypertrophy and heart failure through poorly understood molecular mechanisms. We have recently shown that fibroblast growth factor-21 (FGF21) is produced by the heart and exerts protective effects that prevent cardiac hypertrophy development and oxidative stress. The aim of this study was to determine the effects of FGF21 on the cardiomyopathy associated with obesity development. RESULTS: Fgf21-/- mice showed an enhanced increase in the heart weight/tibia length (HW/TL) ratio in response to the high-fat diet. In keeping with this, echocardiographic measurements confirmed enhanced cardiac hypertrophy in Fgf21-/- mice. At the cellular level, the area of cardiomyocytes was increased in Fgf21-/- mice fed a high-fat diet. Furthermore, a high-fat diet induced fatty acid oxidation in the hearts of Fgf21-/- mice accompanied by an increase in cardiac oxidative stress. Oil-red O staining revealed the presence of higher amounts of lipid droplets in the hearts of Fgf21-/- mice fed a high-fat diet relative to wt mice fed this same diet. Finally, Fgf21-/- mice fed a high-fat diet showed impaired cardiac autophagy and signs of inactive cardiac lipophagy, suggesting that FGF21 promotes autophagy in cardiomyocytes. CONCLUSIONS: Our data indicate that a lack of FGF21 enhances the susceptibility of mice to the development of obesity-related cardiomyopathy. Furthermore, we demonstrate that this cardiac dysfunction is associated with deleterious lipid accumulation in the heart. An impaired ability of FGF21 to promote autophagy/lipophagy may contribute to lipid accumulation and cardiac derangements.

Quiste dermoide cervical. Reporte de caso clínico / Cervical Dermoid cyst. A case report

Los quistes dermoides presentan una incidencia de 1,6 hasta un 6,9 por ciento en la región de cabeza y cuello. Se presenta el siguiente caso clínico con el objetivo de enfatizar en la importancia de un adecuado manejo preoperatorio para establecer una acertada planificación quirúrgica en la exéresis de este tipo de lesiones de la región cervico facial. Se presenta el caso clínico de una paciente femenina de 22 años de edad quien hacía 5 años se notaba un crecimiento redondeado en región submandibular, lo cual le ocasionaba molestias al hablar y ligera afectación de su estética facial. Se decide tratamiento quirúrgico para la extirpación de la lesión quística de gran dimensión. El diagnóstico histopatológico corresponde con un quiste dermoide verdadero. Se concluye que resulta imprescindible realizar un exhaustivo examen físico e imagenológico para lograr resultados satisfactorios en el tratamiento quirúrgico del quiste dermoide cervical(AU)

Dermoid cysts of the head present an incidence from 1.6 to 6.9 percent in the head and neck. A clinical case is presented with the objective of emphasizing the importance of a preoperative management for an adequate surgical planning before operations these kinds of lesions. A, 22 year- old female patient, reported a round lesion in her submandibular area, that increased in size 5 years ago, she also complained of discomfort when she talked and her facial esthetic. We performed a surgical treatment according to the big size of the cyst. Histopathology diagnosis showed truly Dermoid cyst. It can be concluded that it is very important to prepare the patient before the surgical treatment with pertinent laboratory studies, additional physical findings, and imageneologic tests(AU)

Imaging mass spectrometry (IMS) of cortical lipids from preclinical to severe stages of Alzheimer's disease.

Alzheimer's disease (AD) is a progressive neurodegenerative disease affecting millions of patients worldwide. Previous studies have demonstrated alterations in the lipid composition of lipid extracts from plasma and brain samples of AD patients. However, there is no consensus regarding the qualitative and quantitative changes of lipids in brains from AD patients. In addition, the recent developments in imaging mass spectrometry methods are leading to a new stage in the in situ analysis of lipid species in brain tissue slices from human postmortem samples. The present study uses the matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS), permitting the direct anatomical analysis of lipids in postmortem brain sections from AD patients, which are compared with the intensity of the lipid signal in samples from matched subjects with no neurological diseases. The frontal cortex samples from AD patients were classified in three groups based on Braak's histochemical criteria, ranging from non-cognitively impaired patients to those severely affected. The main results indicate a depletion of different sulfatide lipid species from the earliest stages of the disease in both white and gray matter areas of the frontal cortex. Therefore, the decrease in sulfatides in cortical areas could be considered as a marker of the disease, but may also indicate neurochemical modifications related to the pathogenesis of the disease. This article is part of a Special Issue entitled: Membrane Lipid Therapy: Drugs Targeting Biomembranes edited by Pablo V. Escribá.

Consumption of seafood and its estimated heavy metals are associated with lipid profile and oxidative lipid damage on healthy adults from a Spanish Mediterranean area: A cross-sectional study.

The association between the consumption of seafood and its benefits on cardiovascular (CVD) risk can be challenged by its heavy metal (HM) content. This study aimed to explore the association of seafood consumption and its estimated HM contents with the lipid profile and lipid oxidation biomarkers in adults from a Spanish Mediterranean area who do not present risk factors for CVD. In this cross-sectional study, the clinical history, three-day dietary record, lipid profile (LDLc, HDLc, APOB/A, and triglyceride levels), plasma oxidised LDL (oxLDL) and 8-isoprostane levels of 81 adults without risk factors for CVD [43% men, with a mean age of 43.6 years (95%CI: 40.1-47.1)] were assessed. The HM [arsenic (As), cadmium (Cd), mercury (Hg), and lead (Pb)] contents of seafood were estimated according to data from analyses of marine species in the same Mediterranean area. Moderate adherence to the Mediterranean diet (score: 4.6 of 9) with a mean seafood consumption of 74.9g/day (95%CI: 59.9-89.9), including 22.7g of shellfish per day (95%CI: 13.5-31.9), was observed. The estimated HM contents were lower than the provisional tolerable weekly intakes (PTWIs): 21.12µg/kg/week As, 0.57µg/kg/week InAs, 0.15µg/kg/week Cd, 1.11µg/kg/week Hg and 0.28µg/kg/week Pb. After adjusting by confounder variables, an increase in shellfish consumption was associated with increases in the levels of LDLc (P=0.013), non-HDLc (P=0.015), APOB/A (P=0.02) and plasma oxLDL (P=0.002). Moreover, an increase in the estimated As and Hg levels in shellfish was associated with an increase in LDLc (P=0.015 and P=0.018, respectively), non-HDLc (P<0.008 and P<0.008, respectively), APOB/A ratio (P=0.008 and P=0.009, respectively), and oxLDL (P≤0.001 and P≤0.001, respectively) levels. In conclusion, in adults without risk factors for CVD, increasing shellfish consumption, even by a moderate amount, could favour a pro-atherogenic lipid profile and a higher level of oxidised LDL. These associations are likely influenced by the estimated exposure to As and Hg from shellfish despite these values are lower than the PTWIs.

Familial aggregation of schizotypy in schizophrenia-spectrum disorders and its relation to clinical and neurodevelopmental characteristics.

INTRODUCTION: This study explored schizotypy as a familial liability marker for schizophrenia-spectrum disorders (SSD) by examining: 1) the aggregation of schizotypy in families with a SSD patient, 2) whether familial resemblance of schizotypy is associated with ridge dissociations (RD), another SSD liability marker, 3) whether schizotypy aggregation patterns influence patients' psychopathology. METHODS: The sample comprised 30 SSD patients and 82 healthy first-degree relatives. Schizotypy was assessed using the Structured Interview for Schizotypy-Revised (SIS-R). Patients' psychopathology was evaluated using the Comprehensive Assessment of Symptoms and History (CASH). RD were identified as anomalies of the dermal ridge junction. Familiality of SIS-R was investigated using a linear mixed model (LMM) and its strength was assessed using an intraclass correlation coefficient (ICC). Another LMM using the absolute differences in SIS-R scores between all possible pairs of relatives as the dependent variable was fitted to obtain an intra-family resemblance score, a family-specific indicator of resemblance of SIS-R scores within each family. RESULTS: 1) Schizotypy was familial (ICC = 0.30); families with high resemblance displayed low schizotypy, whereas families with low resemblance included at least one healthy relative with high schizotypy (p < 0.001). 2) Relatives with RD had higher SIS-R scores (p = 0.018) and belonged to families with discordant schizotypy scores among members (p < 0.001). 3) Patients from high schizotypy families showed more severe disorganized symptoms at the psychotic episode (p = 0.035) and 1 year later (p = 0.011). CONCLUSIONS: Schizotypy is a marker of vulnerability for SSD that runs within a subgroup of families. The schizotypy familial aggregation pattern correlates with RD in relatives and with patients' psychopathology.

Thermogenic activation represses autophagy in brown adipose tissue.

BACKGROUND: Brown adipose tissue (BAT) thermogenesis is an adaptive process, essential for energy expenditure and involved in the control of obesity. Obesity is associated with abnormally increased autophagy in white adipose tissue. Autophagy has been proposed as relevant for brown-vs-white adipocyte differentiation; however, its role in the response of BAT to thermogenic activation is unknown. METHODS: The effects of thermogenic activation on autophagy in BAT were analyzed in vivo by exposing mice to 24 h cold condition. The effects of norepinephrine (NE), cAMP and modulators of lysosomal activity were determined in differentiated brown adipocytes in the primary culture. Transcript expression was quantified by real-time PCR, and specific proteins were determined by immunoblot. Transmission electron microscopy, as well as confocal microscopy analysis after incubation with specific antibodies or reagents coupled to fluorescent emission, were performed in BAT and cultured brown adipocytes, respectively. RESULTS: Autophagy is repressed in association with cold-induced thermogenic activation of BAT in mice. This effect was mimicked by NE action in brown adipocytes, acting mainly through a cAMP-dependent protein kinase A pathway. Inhibition of autophagy in brown adipocytes leads to an increase in UCP1 protein and uncoupled respiration, suggesting a repressing role for autophagy in relation to the activity of BAT thermogenic machinery. Under basal conditions, brown adipocytes show signs of active lipophagy, which is suppressed by a cAMP-mediated thermogenic stimulus. CONCLUSIONS: Our results show a noradrenergic-mediated inverse relationship between autophagy and thermogenic activity in BAT and point toward autophagy repression as a component of brown adipocyte adaptive mechanisms to activate thermogenesis.

Evaluation of the effectiveness of three physiotherapeutic treatments for subacromial impingement syndrome: a randomised clinical trial.

OBJECTIVE: To determine whether dexketoprofen administered by phonophoresis or iontophoresis is more effective for the treatment of subacromial impingement syndrome (SIS) than conventional ultrasound therapy. DESIGN: Randomised clinical trial. SETTING: University hospital. PARTICIPANTS: Ninety-nine participants with SIS without a complete tear of the rotator cuff were assigned at random to three intervention groups. INTERVENTION GROUPS: Participants received ultrasound (n=32), phonophoresis with dexketoprofen (50mg/session) (n=33) or iontophoresis with dexketoprofen (50mg/session) (n=34). All participants completed 20 treatment sessions plus exercise therapy and cryotherapy. OUTCOME MEASURES: A visual analogue scale (VAS), the Constant-Murley Scale (CMS) and the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire were administered pre-treatment (baseline), post-treatment and 1 month post-treatment. RESULTS: At baseline, there were no differences between the groups. Post-treatment, VAS score improved by -1.2 points and CMS score improved by 8.9 points in the ultrasound group compared with the iontophoresis group [95% confidence interval (CI) -0.2 to -2.2 and 95% CI 17.0 to 0.7, respectively]. CMS score improved by 7.1 points in the phonophoresis group compared with the iontophoresis group (95% CI 14.8 to -0.7). At 1 month post-treatment, no significant differences were detected between the groups. VAS, CMS and DASH scores of all groups improved post-treatment and at 1 month post-treatment. CONCLUSION: Ultrasound, iontophoresis with dexketoprofen and phonophoresis with dexketoprofen can improve pain, shoulder function, and physical functioning and symptoms in the upper limb in patients with SIS without a complete tear of the rotator cuff. CLINICAL TRIALS. GOV REGISTRATION NUMBER: NCT01748188.

The effect of snacking and eating frequency on dietary quality in British adolescents.

PURPOSE: To describe the effects of number of eating occasions and snacks on dietary quality (DQ), defined as adherence to dietary recommendations. METHODS: A sample of 884 adolescents (11-18 years) in the UK National Diet and Nutrition Survey (NDNS) were included. The Diet Quality Index for Adolescents (DQI-A) was implemented. The total number of eating occasions and snacks was frequency of food or beverages consumed over 24 h and frequency of foods or beverages consumed outside of the three mealtimes, respectively. Results were generated with and without low-energy food under 210 kJ (50 kcal). Regression models were generated with DQ score as the outcome variable and number of eating occasions and snacks as predictors. RESULTS: The mean (95 % CI) DQ score was 31.1 % (30.2, 32.0). The mean number of eating occasions and snacks was 7.5 (7.3, 7.7) and 2.6 (2.6, 2.7) times/day, respectively. When low-energy events were excluded, the mean number of eating occasions and snacks reduced to 6.2 (6.1, 6.4) and 2.0 (2.0, 2.1) times/day, respectively. DQ score increased by 0.74 points (0.42, 1.05; p < 0.01) and 0.55 points (-0.08, 0.69; p = 0.17) for total eating occasions and snacks, respectively. When low-energy events were excluded, DQ score increased by 0.30 points (-0.84, 0.69; p = 0.13) for each eating occasion and decreased by 1.20 points (-2.1, -0.3; p < 0.01) for each snack. CONCLUSION: Eating more frequently improves dietary quality especially if some eating occasions are low in energy. A focus on replacing high-energy snacks with low-energy alternatives rather than reducing the number of eating occasions may result in improved dietary quality in adolescents.

C/EBPß is required in pregnancy-induced cardiac hypertrophy.

AIM: Pregnancy is a physiological model of adaptive and reversible heart enlargement, but the molecular mechanisms determining this kind of physiologic cardiac hypertrophy are poorly known. Here, we analyzed the role of the transcription factor C/EBPß in the development of pregnancy-induced cardiac hypertrophy. RESULTS: C/EBPß+/- mice at day 18 of gestation were used as happloinsufficiency model of late pregnancy. We found that C/EBPß expression was specifically increased in hearts from Wt pregnant mice whereas expression of other C/EBP subtypes (α and δ) was not affected by gestation. Pregnancy-induced changes in systemic metabolic and hormonal profiles were not essentially different in Wt versus C/EBPß+/- mice. However, C/EBPß+/- mice developed pregnancy-induced heart hypertrophy to a lower extent relative to Wt mice. Furthermore, hearts from C/EBPß+/- mice have alterations in fatty acid oxidation genes and reductions in the expression levels of glucose transporters that may compromise metabolic cardiac function during pregnancy. Among marker genes of inflammation, interleukin-6 (Il-6) showed a marked differential behavior in C/EBPß+/- pregnant mice: pregnancy strongly induced cardiac Il-6 expression in wt, a phenomenon that did not occur in C/EBPß+/- mice. Moreover, marker genes for M2 macrophages were decreased in C/EBPß+/- pregnant mice and in C/EBPß-/- mice subjected to LPS stimulus. CONCLUSIONS: Here we found that normal levels of C/EBPß are required for hypertrophy development during pregnancy. Events such as the increase in IL-6 in the heart of pregnant mice are prevented in C/EBPß+/- animals. Moreover, C/EBPß controls M2-macrophage gene expression in the heart. Thus, C/EBPß appears as a transcription factor required for cardiac hypertrophy response to gestation.

Effects of galanin subchronic treatment on memory and muscarinic receptors.

The cholinergic pathways, which originate in the basal forebrain and are responsible for the control of different cognitive processes including learning and memory, are also regulated by some neuropeptides. One of these neuropeptides, galanin (GAL), is involved in both neurotrophic and neuroprotective actions. The present study has evaluated in rats the effects on cognition induced by a subchronic treatment with GAL by analyzing the passive avoidance response, and the modulation of muscarinic cholinergic receptor densities and activities. [(3)H]-N-methyl-scopolamine, [(3)H]-oxotremorine, and [(3)H]-pirenzepine were used to quantify the density of muscarinic receptors (MRs) and the stimulation of the binding of guanosine 5'-(γ-[(35)S]thio)triphosphate by the muscarinic agonist, carbachol, to determine their functionality. Some cognitive deficits that were induced by the administration of artificial cerebrospinal fluid (aCSF) (i.c.v. aCSF 2 µl/min, once a day for 6 days) were not observed in the animals also treated with GAL (i.c.v. 1.5 mmol in aCSF, 2 µl/min, once a day for 6 days). GAL modulates the changes in M1 and M2 MR densities observed in the rats treated with aCSF, and also increased their activity mediated by G(i/o) proteins in specific areas of the dorsal and ventral hippocampus. The subchronic administration of the vehicle was also accompanied by an increased number of positive fibers and cells for GAL around the cortical tract of the cannula used, but that was not the case in GAL-treated rats. In addition, the increase of GAL receptor density in the ventral hippocampus and entorhinal cortex in the aCSF group was avoided when GAL was administered. The number of acetylcholinesterase (AChE)-positive neurons was decreased in the nucleus basalis of Meynert of both GAL- and aCSF-treated animals. In summary, GAL improves memory-related abilities probably through the modulation of MR density and/or efficacy in hippocampal areas.

Neurotransmitter receptor localization: from autoradiography to imaging mass spectrometry.

Autoradiography is used to determine the anatomical distribution of biological molecules in human tissue and experimental animal models. This method is based on the analysis of the specific binding of radiolabeled compounds to locate neurotransmitter receptors or transporters in fresh frozen tissue slices. The anatomical resolution obtained by quantification of the radioligands has allowed the density of receptor proteins to be mapped over the last 40 years. The data yielded by autoradiography identify the receptors at their specific microscopic localization in the tissues and also in their native microenvironment, the intact cell membrane. Furthermore, in functional autoradiography, the effects of small molecules on the activity of G protein-coupled receptors are evaluated. More recently, autoradiography has been combined with membrane microarrays to improve the high-throughput screening of compounds. These technical advances have made autoradiography an essential analytical method for the progress of drug discovery. We include the future prospects and some preliminary results for imaging mass spectrometry (IMS) as a useful new method in pharmacodynamic and pharmacokinetic studies, complementing autoradiographic studies. IMS results could also be presented as density maps of molecules, proteins, and metabolites in tissue sections that can be identified, localized, and quantified, with the advantage of avoiding any labeling of marker molecules. The limitations and future developments of these techniques are discussed here.

Opposite alterations in FGF21 and FGF19 levels and disturbed expression of the receptor machinery for endocrine FGFs in obese patients.

OBJECTIVE: Fibroblast growth factor (FGF)-21, and possibly FGF19, protect against type 2 diabetes mellitus (T2DM) and obesity in rodents. We investigated the circulating levels of FGF21 and FGF19 in obese patients with varying degrees of abnormal glucose homeostasis, and we determined gene expression for FGF receptors (FGFR1-4) and the co-receptor ß-Klotho, in liver and adipose tissues. SUBJECTS AND METHODS: We analyzed 35 lean healthy (71% men) and 61 obese patients (49% men, median body mass index (BMI): 40.5 kg m(-2), interquartile range: 34.7-46.2). Among obese patients, 36 were normoglycemic, 15 showed impaired glucose tolerance and 10 had T2DM. Biopsies from liver and visceral and subcutaneous fat from a subset of obese patients and controls were analyzed. FGF19 and FGF21 levels were measured using enzyme-linked immunosorbent assay, and tissue mRNA and protein levels by reverse transcription-polymerase chain reaction and immunoblotting. RESULTS: FGF21 serum levels were significantly increased in obese patients compared with controls (P<0.001), whereas FGF19 levels were decreased (P < 0.001). FGF21 levels were positively correlated with homeostasis model assessment of insulin resistance (P = 0.0002, r = 0.37) and insulin (P = 0.001, r = 0.32), whereas FGF19 levels were negatively correlated (P = 0.007, r = -0.27; P=0.003, r = -0.28; respectively). After adjusting for BMI, the correlations of FGF21 and FGF19 levels with indicators of abnormal glucose homeostasis were not significant. In obese patients, the hepatic expression of FGF21 was increased. (P = 0.04). ß-Klotho transcript levels in visceral fat (P = 0.002) and ß-Klotho protein levels in subcutaneous (P = 0.03) and visceral fat (P = 0.04) were significantly reduced in obese patients, whereas hepatic levels for ß-Klotho (P = 0.03), FGFR1 (P = 0.04) and FGFR3 (P = 0.001) transcripts were significantly increased. CONCLUSIONS: Obesity is characterized by reciprocal alterations in FGF19 (decrease) and FGF21 (increase) levels. Although worsened in diabetic obese patients, obesity itself appears as the predominant determinant of the abnormalities in FGF21 and FGF19 levels. Opposite changes in ß-Klotho expression in fat and liver indicate potential tissue-specific alterations in the responsiveness to endocrine FGFs in obesity.

Galanin activated Gi/o-proteins in human and rat central nervous systems.

The neuropeptide galanin (GAL) is involved in the control of hormone secretion, nociception, feeding behavior, attention, learning and memory. The anatomical localization of galanin receptors in the brain has been described using autoradiography and immunohistochemistry, but both techniques are limited by the availability of specific radioligands or antibodies. Functional autoradiography provides an alternative method by combining anatomical resolution and information of the activity mediated by G-protein coupled receptors. The present study analyzes the functional GAL receptors coupled to Gi/o-proteins in human and rat brain nuclei using [(35)S]GTPγS autoradiography. The results show the anatomical distribution of Gi/o-proteins activated by GAL receptors that trigger intracellular signaling mechanisms. The activity mediated by GAL receptors in human and rat brain showed a good correlation of the net stimulation in areas such as spinal cord, periaqueductal gray, putamen, CA3 layers of hippocampus, substantia nigra and diverse thalamic nuclei. The functional GAL receptors coupled to Gi/o-proteins showed a similar pattern for both species in most of the areas analyzed, but some discrete nuclei showed differences in the activity mediated by GAL, such as the ventroposteromedial thalamic nucleus, or areas that regulate learning and memory processes in the hippocampus. Taken into consideration the present results, the rat could be used as an experimental model for the study of the physiological role of GAL-mediated neurotransmission and the modulation of GAL receptors activity in the human CNS.

Type-1 cannabinoid receptor activity during Alzheimer's disease progression.

The activity of CB1 cannabinoid receptors was studied in postmortem brain samples of Alzheimer's disease (AD) patients during clinical deterioration. CB1 activity was higher at earlier AD stages in limited hippocampal areas and internal layers of frontal cortex, but a decrease was observed at the advanced stages. The pattern of modification appears to indicate initial hyperactivity of the endocannabinoid system in brain areas that lack classical histopathological markers at earlier stages of AD, indicating an attempt to compensate for the initial synaptic impairment, which is then surpassed by disease progression. These results suggest that initial CB1 stimulation might have therapeutic relevance.

Interleukin-6 deletion in mice driven by aP2-Cre-ERT2 prevents against high-fat diet-induced gain weight and adiposity in female mice.

AIM: Interleukin-6 (IL-6) is a major cytokine controlling body weight and metabolism, but because many types of cells can synthesize and respond to IL-6 considerable uncertainty still exists about the mechanisms underlying IL-6 effects. Therefore, the aim of this study was to analyse the effects of tissue-specific deletion of IL-6 using a fatty acid binding protein (aP2) promoter-Cre inducible system (aP2-Cre-ERT2). METHODS: Tissue-specific IL-6 KO mice (aP2-IL-6 KO mice) were produced upon tamoxifen administration and were fed a high-fat diet (HFD, 58.4% kcal from fat) or a control diet (18%) for 14 weeks. RESULTS: aP2-IL-6 KO female mice on a HFD gained less weight and adiposity than littermate wild-type mice, but these effects were not observed in males. Hypothalamic factors such as NPY and AgRP showed a pattern of expression consistent with this sex-specific phenotype. PGC-1α expression was increased in several tissues in aP2-IL-6 KO female mice, which is compatible with increased energy expenditure. Serum leptin, insulin, glucose, cholesterol and triglycerides levels were increased by HFD, and in females IL-6 deficiency reversed this effect in the case of insulin and cholesterol. HFD induced impaired responses to insulin and glucose tolerance tests, but no significant differences between genotypes were observed. CONCLUSION: The present results demonstrate that deletion of IL-6 driven by aP2-Cre regulates body weight, body fat and metabolism in a sex-specific fashion.

Alarmin high-mobility group B1 (HMGB1) is regulated in human adipocytes in insulin resistance and influences insulin secretion in ß-cells.

BACKGROUND: The nuclear protein high-mobility group box 1 (HMGB1) can be passively released by necrotic cells or secreted actively by several cell types to regulate immune and inflammatory responses, as well as tissue remodeling. We herein aimed to characterize the effect of insulin resistance on HMGB1 in adipose tissue and to examine its potential role as a metabolic regulator in ß-pancreatic cells. DESIGN: Plasma HMGB1 concentration and adipose HMGB1 expression were assessed in relation to obesity and insulin resistance. Cultured adipocytes from lean and obese patients were used to investigate the intracellular distribution and factors regulating HMGB1 release, as well as to test its effects on adipogenesis and lipid metabolism. A regulatory role for HMGB1 in insulin secretion was also investigated. RESULTS: Circulating HMGB1 was positively associated with body mass index, while adipose HMGB1 mRNA levels correlated with the expression of inflammatory markers. Insulin resistance modified the intracellular distribution of HMGB1 in human adipocytes, with HMGB1 being predominantly nuclear in lean and obese normoglycemic individuals while localized to the cytosol in obese patients with type 2 diabetes. Adipocytes from lean individuals exposed to conditioned media from lipopolysaccharide-stimulated macrophages induced HMGB1 redistribution to the cytoplasm and release. HMGB1 treatment had no effect on differentiation and lipid metabolism in adipocytes. However, HMGB1, whose circulating levels correlated with postload insulin concentration, increased both insulin release and intracellular Ca(2+) concentration in INS-1 cells. CONCLUSIONS: These findings show, for the first time, that HMGB1 expression and release by human adipocytes is altered by inflammatory conditions as those imposed by obesity and insulin resistance. Our data reveal a novel role for HMGB1 as a stimulatory factor of insulin secretion of ß-pancreatic cells.

Oxidative stress in immunocompetent patients with severe community-acquired pneumonia. A pilot study

OBJECTIVE: A comparison was made of the oxidative stress (OS) levels of patients with either viral or bacterial severe community-acquired pneumonia (sCAP) and of patients without infection (healthy volunteers (HV) and patients with acute myocardial infarction (AMI)). DESIGN: A prospective observational study was made. PATIENTS: Critically ill patients with sCAP. VARIABLES: The TBARS level was measured as an index of oxidative injury. SOD, CAT and redox glutathione system (GSH, GSSG, GR, GPx) activities were measured as reflecting antioxidant capacity. Severity of illness was assessed by the APACHE II, SOFA and SIRS scores. RESULTS: Thirty-seven subjects were included: 15 patients with CAP (12 of bacterial origin [BCAP] and 3 due to 2009 A/H1N1 virus [VCAP]), 10 HV and 12 AMI patients. Intensive care CAP mortality was 26.7% (n = 4). Plasmatic TBARS levels were higher in CAP patients than in HV, but similar to those recorded in AMI patients. In contrast, VCAP was associated with lower TBARS levels, and some components of the glutathione redox system were higher in BCAP patients and HV. The OS levels did not differ between survivors and non-survivors. CONCLUSION: Our results suggest the occurrence of higher OS in sCAP patients compared with HV. In contrast, lower TBARS levels were observed in VCAP patients, suggesting an increase of antioxidant activity related to the redox glutathione system. However, further research involving a larger cohort is needed in order to confirm these findings

OBJETIVOS: Comparar el estrés oxidativo (EO) en pacientes con neumonía comunitaria grave (NCG) según su etiología y respecto de voluntarios sanos (VS) y pacientes con infarto agudo de miocardio (IAM). DISEÑO: Estudio prospectivo, observacional. PACIENTES: Pacientes con NCG ingresados en unidades de cuidados intensivos. Variables Los niveles de lipoperoxidación (TBARS) fueron considerados como índice de oxidación, mientras que SOD, CAT y la actividad del sistema redox- glutation (GSH, GSSG, GR, GPx) fueron considerados capacidad antioxidante. La gravedad de los pacientes fue valorada mediante las escalas APACHE II, SOFA y SIRS. RESULTADOS: Treinta y siete sujetos fueron incluidos, 15 pacientes con NCG (12 con etiología bacteriana [NB] y 3 viral 2009 A/H1N1 [NV]), 10 VS y 12 con IAM. La mortalidad global fue del 26,7% (n = 4). Los TBARS plasmáticos fueron superiores en NCG respecto de VS, pero similares al IAM. En contraste, la NV se asoció con menores niveles de TBARS e incremento de componentes del sistema redox-glutation respecto de NB y voluntarios sanos. No se observó asociación entre mortalidad y EO. CONCLUSIÓN: Nuestros resultados evidencian la presencia de EO en pacientes con NCG respecto de los controles. En contraste, la evidencia de un menor nivel de TBARS en la NV respecto de los VS sugiere un incremento de la actividad antioxidante relacionada con el sistema redox-glutation. Sin embargo, son necesarias nuevas investigaciones para confirmar estos hallazgos

Oxidative stress in immunocompetent patients with severe community-acquired pneumonia. A pilot study.

OBJECTIVE: A comparison was made of the oxidative stress (OS) levels of patients with either viral or bacterial severe community-acquired pneumonia (sCAP) and of patients without infection (healthy volunteers (HV) and patients with acute myocardial infarction (AMI)). DESIGN: A prospective observational study was made. PATIENTS: Critically ill patients with sCAP. VARIABLES: The TBARS level was measured as an index of oxidative injury. SOD, CAT and redox glutathione system (GSH, GSSG, GR, GPx) activities were measured as reflecting antioxidant capacity. Severity of illness was assessed by the APACHE II, SOFA and SIRS scores. RESULTS: Thirty-seven subjects were included: 15 patients with CAP (12 of bacterial origin [BCAP] and 3 due to 2009 A/H1N1 virus [VCAP]), 10 HV and 12 AMI patients. Intensive care CAP mortality was 26.7% (n=4). Plasmatic TBARS levels were higher in CAP patients than in HV, but similar to those recorded in AMI patients. In contrast, VCAP was associated with lower TBARS levels, and some components of the glutathione redox system were higher in BCAP patients and HV. The OS levels did not differ between survivors and non-survivors. CONCLUSION: Our results suggest the occurrence of higher OS in sCAP patients compared with HV. In contrast, lower TBARS levels were observed in VCAP patients, suggesting an increase of antioxidant activity related to the redox glutathione system. However, further research involving a larger cohort is needed in order to confirm these findings.