Lilian Marques Silva

Leptin resistance and desensitization of hypophagia during prolonged inflammatory challenge

Acute exposure to bacterial lipopolysaccharide (LPS) is a potent inducer of immune response as well as hypophagia. Nevertheless, desensitization of responses to LPS occurs during long-term exposure to endotoxin. We induced endotoxin tolerance, injecting repeated (6LPS) LPS doses compared with single (1LPS) treatment. 1LPS, but not 6LPS group, showed decreased food intake and body weight, which was associated with an increased plasma leptin and higher mRNA expression of OB-Rb, MC4R, and SOCS3 in the hypothalamus. Hypophagia induced by 1LPS was associated with lower levels of 2-arachidonoylglycerol (2-AG), increased number of p-STAT3 neurons, and decreased AMP-activated protein kinase (AMPK) activity. Desensitization of hypophagia in the 6LPS group was related to high 2-AG, with no changes in p-STAT3 or increased p-AMPK. Leptin decreased food intake, body weight, 2-AG levels, and AMPK activity and enhanced p-STAT3 in control rats. However, leptin had no effects on 2-AG, p-STAT3, or p-AMPK in the 1LPS and 6LPS groups. Rats treated with HFD to induce leptin resistance showed neither hypophagia nor changes in p-STAT3 after 1LPS, suggesting that leptin and LPS recruit a common signaling pathway in the hypothalamus to modulate food intake reduction. Desensitization of hypophagia in response to repeated exposure to endotoxin is related to an inability of leptin to inhibit AMPK phosphorylation and 2-AG production and activate STAT3. SOCS3 is unlikely to underlie this resistance to leptin signaling in the endotoxin tolerance. The present model of prolonged inflammatory challenge may contribute to further investigations on mechanisms of leptin resistance.

Glucocorticoids are required for meal-induced changes in the expression of hypothalamic neuropeptides.

Glucocorticoid deficiency is associated with a decrease of food intake. Orexigenic peptides, neuropeptide Y (NPY) and agouti related protein (AgRP), and the anorexigenic peptide proopiomelanocortin (POMC), expressed in the arcuate nucleus of the hypothalamus (ARC), are regulated by meal-induced signals. Orexigenic neuropeptides, melanin-concentrating hormone (MCH) and orexin, expressed in the lateral hypothalamic area (LHA), also control food intake. Thus, the present study was designed to test the hypothesis that glucocorticoids are required for changes in the expression of hypothalamic neuropeptides induced by feeding. Male Wistar rats (230-280 g) were subjected to ADX or sham surgery. ADX animals received 0.9% NaCl in the drinking water, and half of them received corticosterone in the drinking water (B: 25 mg/L, ADX+B). Six days after surgery, animals were fasted for 16 h and they were decapitated before or 2 h after refeeding for brain tissue and blood collections. Adrenalectomy decreased NPY/AgRP and POMC expression in the ARC in fasted and refed animals, respectively. Refeeding decreased NPY/AgRP and increased POMC mRNA expression in the ARC of sham and ADX+B groups, with no effects in ADX animals. The expression of MCH and orexin mRNA expression in the LHA was increased in ADX and ADX+B groups in fasted condition, however there was no effect of refeeding on the expression of MCH and orexin in the LHA in the three experimental groups. Refeeding increased plasma leptin and insulin levels in sham and ADX+B animals, with no changes in leptin concentrations in ADX group, and insulin response to feeding was lower in this group. Taken together, these data demonstrated that circulating glucocorticoids are required for meal-induced changes in NPY, AgRP and POMC mRNA expression in the ARC. The lower leptin and insulin responses to feeding may contribute to the altered hypothalamic neuropeptide expression after adrenalectomy.

Use of Electroless Plating Copper Thin Films for Catalysis

Recently, it was demonstrated that copper thin films show good adsorption characteristics for organic polar and non-polar compounds. Also, these films when used in small cavities can favor preconcentration of these organic compounds. It is also known that copper oxide can provide catalysis of organic compounds. Therefore, the aim of this work is the study of copper thin film catalysis when used in small cavities. Copper thin films, 25 nm thick, were deposited on silicon and/or rough silicon. These films do not show oxide on the surface when analyzed by Rutherford backscattering. Also, Raman analysis of these films showed only silicon bands, due to the substrate, however infrared spectroscopy shows oxide bands for films exposed to organic compound aqueous solutions. Cavities with copper films deposited inside were tested with a continuous flow of n-hexane, acetone or 2-propanol admitted in the system. The effluent was analyzed by Quartz Crystal Microbalance. It was shown that n-hexane or acetone can be trapped. The system also shows good reproducibility. Tests of catalysis were carried out using Raman spectroscopy and heating the films up to 300°C during 3 minutes after exposure to n-hexane, 2- propanol and acetophenone – pure or saturated aqueous solution. After the exposure, Raman spectra present intense bands only for 2-propanol, indicating that adsorption easily occurs. However, after heating with all solutions it was not found only silicon bands. Raman microscopy after heating also showed copper oxide cluster formation and, eventually, graphite formation. Although the heating provides oxide copper formation, this reaction does not produce a high amount of residues, which means that catalysis is possible in this condition. Thus, a simple device using copper thin films can be useful as sample pretreatment on microTAS development.

Low cost microstructures for preconcentration of polar and non-polar organic compounds

Films produced by plasma polymerization of ethyl ether and methyl or ethyl acetate show good adsorption characteristic for polar and non-polar organic compounds. These films when used in microchannels machined in a 3D-structure present some preconcentration of organic compounds. Therefore, the aim of this work is to investigate the physical-chemical preconcentration mechanisms on this structure. The test molecules used were n-hexane and 2-propanol. Quartz crystal microbalance and mass spectrometry were used to measure preconcentration. Two different procedures for reactant injection on the structure were used: a continuous flow during several minutes or a small amount injected on a single pulse and in a few seconds. The microchannels were also modified by the introduction of small ceramic particles for enhancement of the flow dispersion. It was possible to notice for all films a similar kinetic of retention. The main removal mechanism is adsorption. Although all films can provide the removal of the adsorbents molecules, the most important characteristic for the adsorption and/or retention is the surface condition. Thus, the retention of polar compound can be troubled if a non-polar compound was used previously. The most promising films for retention are ethyl ether and ethyl acetate when n-hexane and 2-propanol are used as test molecules. The results using n-hexane or 2-propanol point out the use of low-cost microchannels for preconcentration development.

Excessive training induces molecular signs of pathologic cardiac hypertrophy: DA ROCHA et al.

Chronic exercise induces cardiac remodeling that promotes left ventricular hypertrophy and cardiac functional improvement, which are mediated by the mammalian or the mechanistic target of rapamycin (mTOR) as well as by the androgen and glucocorticoid receptors (GRs). However, pathological conditions (i.e., chronic heart failure, hypertension, and aortic stenosis, etc.) also induce cardiac hypertrophy, but with detrimental function, high levels of proinflammatory cytokines and myostatin, elevated fibrosis, reduced adenosine monophosphate‐activated protein kinase (AMPK) activation, and fetal gene reactivation. Furthermore, recent studies have evidenced that excessive training induced an inflammatory status in the serum, muscle, hypothalamus, and liver, suggesting a pathological condition that could also be detrimental to cardiac tissue. Here, we verified the effects of three running overtraining (OT) models on the molecular parameters related to physiological and pathological cardiac hypertrophy. C57BL/6 mice performed three different OT protocols and were evaluated for molecular parameters related to physiological and pathological cardiac hypertrophy, including immunoblotting, reverse transcription polymerase chain reaction, histology, and immunohistochemistry analyses. In summary, the three OT protocols induced left ventricle (LV) hypertrophy with signs of cardiac fibrosis and negative morphological adaptations. These maladaptations were accompanied by reductions in AMPKalpha (Thr172) phosphorylation, androgen receptor, and GR expressions, as well as by an increase in interleukin‐6 expression. Specifically, the downhill running–based OT model reduced the content of some proteins related to the mTOR signaling pathway and upregulated the β‐isoform of myosin heavy‐chain gene expression, presenting signs of LV pathological hypertrophy development.

Large Surface Area of HMDS Plasma Polymerized Thin Film Used for Production of Miniaturized Structures

The aim of this work was the production of a large surface area of hexamethyldisilazane (HMDS) plasma-deposited thin films and their applicability in a miniaturized structure useful for preliminary analysis of organic mixtures. The HMDS plasma films were produced with different surface areas and morphologies, and all films adsorbed polar and non-polar organic compounds. A low cost miniaturized structure was manufactured in glass using a Milling cutter and covered with HMDS plasma films. Good agreement was observed between simulation and experimental results on those microstructures. The observed different performance between pure and mixtures of organic compound samples suggests that the proposed system is a simple setup that could be useful for rough analysis of a fuel.

Perfluorocompound and Hexamethyldisilazane Thin Film Composite Material Used for Surface Modification

The aim of this work was to produce, to characterize and test selective membranes based on derivatives from organic fluorinated/silicon compounds. The produced composites presented silicon and fluorinated species on the surface. The morphology of these non uniform surfaces showed big domains in the micrometer scale but a closer view reveals structures also in the nanometer range. Contact angle measurements showed a mildly hydrophobic and organophilic surface. Polymeric tapes treated with this composite showed an increase on permeation rate for the organic compounds. An acrylic device that employed this modified polymeric tape was manufactured and used for sample pretreatment during chemical analysis.