Differentiation of human pre-adipocytes by recombinant adiponectin

Obesity has become a global health problem and it is linked to a higher risk of diseases and metabolic disorders such as diabetes, cardiovascular disease, and cancer. The adipose tissue plays an important role in monitoring and controlling whole-body metabolism by secreting a variety of bioactive molecules such as adiponectin. Deficiencies of this hormone can cause type II diabetes and cardiovascular disease both in mice and in humans. Therefore, adiponectin is an attractive molecule to use in human therapy, particularly in a recombinant form. The source of recombinant adiponectin could be the expression of full-length adiponectin either in Escherichia coli, or in baculovirus. In this work we express and purify human adiponectin in both systems. The adiponectin produced by baculovirus was found to be 10 times more active as far as oligomerization and human pre-adipocyte differentiation are concerned, when compared with adiponectin produced by E. coli. We presume that adiponectin expressed in baculovirus has post-translation modifications not made by bacteria which may be responsible for these differences in activity. This renders adiponectin produced by baculovirus a better candidate for the treatment of type II diabetes and cardiovascular disease.Instituto Gulbenkian de CiênciaFundação para a Ciência e a Tecnologia (FCT

Dyeing of PA6.6 fibers - Effect of solvent and temperature on thermal properties

Dyeing fibers at low temperatures has many advantages such as savings in energy and avoiding alterations to the physical properties of the fibers being dyed or other fibers also present in blends, The problem of low temperature dyeing in synthetic fibers is that it difficults the dye diffusion into the fiber. In the case of polyamide 6.6 microfibers, by using benzyl alcohol as an auxiliary dyeing, it was shown that good diffusion was obtained for the dye exhaustion with metal complex dyes at temperatures more than 30°C below the normal dyeing temperature for the dye exhaustion with metal complex dyes. Using thermal analysis methods these results were shown to be caused by the lowering of the T g of the fiber when in the presence of benzyl alcohol.Fundação para a Ciência e a Tecnologia (FCT

Application of a fungal extract with laccase activity to improve the enzymatic hydrolysis of eucalyptus bark residues

This work had the financial support from the Portuguese Foundation for Science and Technology under the scope of Project EcoTech (POCI-01-0145-FEDER-032206). The authors also acknowledge RAIZ for kindly providing the residues of eucalyptus bark.info:eu-repo/semantics/publishedVersio

Metais pesados em agregados de solo submetido à aplicação sucessiva de lodo de esgoto.

RESUMO: A aplicação de lodo no solo é uma importante alternativa de reciclagem desse tipo de resíduo, de forma a manter os níveis de fertilidade do solo e promover a produtividade competitiva das culturas. No entanto , a presença de metais pesados pode limitar seu uso , sendo que o presente trabalho teve por objetivo avaliar os teores totais de alguns metais pesados e sua distribuição em classes de agregado em função da aplicação sucessiva de lodo de esgoto num solo agrícola . O estudo foi realizado no Campo Experimental da Embrapa Meio Ambiente - Jaguariúna (SP) e foram analisados 3 tratamentos: controle, adubação mineral e 1N (10 t ha - 1 ano - 1 de lodo em base seca ), na profundidade 0 -5 cm. O solo coletado foi passado empeneira de 9 ,5 2 mm de abertura e fracionado em agregados com dimensões: entre 9,52 - 4,00, entre 4,00 - 2,00mm, entre 2,00 - 0,25mm, entre 0,25 - 0,053mm, e menor que 0,053mm. Os teores totais de metais pesados do solo intacto e das classes de agregado foram quantificados por ICP-OES após extração seguindo procedimento da USEPA 3051. Não houve diferença significativa dos tratamentos na distribuição dos agregados do solo por classe de diâmetro. O lodo de esgoto elevou os teores de Cu, Ni e Zn. porém apresentaram baixos teores quando ponderados pela massa. A aplicação do lodo proporcionou aumento dos teores de metais principalmente nas classes de agregado 2,00-0,25mm e 0,25-0,053mm

Using mass spectrometry techniques for the elucidation of structural features of immunostimulatory polysaccharides from medicinal plantsaqueous extracts

Polysaccharides are well known for their immunostimulatory properties, which are closely related to structural fèatures, such as molecular weight, branching, acetylation degree and acetylation pattem, and presence of speciflc structures [1, 2]. TTie electrospray ionization-mass spectrometry (ESI-MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) have shown to be powerfül techniques that can provide detailed information regarding the structural features of polysaccharides and help to clarify its relation to some ofthe observed immunostimulatory properties [3-6]. TTie present work desenhes the use of ESI-MS and ESI-MS/MS in the confimation and elucidation of structural features of immunostimulatory polysaccharides present in lhe aqueous extracts of Fraxinus angustifolia dned leaws and Ptemapaitum tridentatum dried inflorescences. The polysaccharides present in the aqueous extracts of F. angustifolia dried leaves and P. fridentafum dried inflorescences were isolated and fractionated using dialysis, ethanol precipitation, and anionexchange chromatography. Sugar and linkage analysis suggested that the aqueous extracts of F. angustifolia were mostly composed of pectic polysaccharides, while those from P. tridentatum contained a mixture of pectic polysacchahdes, galactomannans, and also xyloglucans. Selected fractions isolated from lhe aqueous extracts of F. angustifolia and P. tridentatum evidenced an in vitro macrophage immunostimulatory activity, expressed through the production of nitric oxide (NO). Enzymatic hydrolysis of the isolated polysaccharides, fbllowed by size exclusion chromatography of the resulting oligosaccharides (OS), and ESI-MS and ESI-MS/MS characterization allowed to confirm the presence of pectic polysaccharides, possibly with xylogalaturonan domains, in the aqueous extracts of F. angustifolia. For the aqueous extracts of P. fridentatum it was possible to detect the presence of galactomannans acetylated at 0-2. Moreover, it was possible to identify acetylated OS that presented an acetylation pattem with much more acetyl groups than those observed for galactomannans from coffee infusions [3]. The saponification ofthe fraction isolated from the aqueous extracts of P. tridentatum dried inflorescences, which contained the acetylated galactomannans, caused a decrease in the NO macrophage production, confirming lhe galactomannans acetylation as an important structural feature for lhe expression of the in vitm macrophage immunostimulatory activity previously registered.info:eu-repo/semantics/publishedVersio

Btk Mutations Selectively Regulate Btk Expression And Upregulate Monocyte Xbp1 Mrna In Xla Patients.

Mutations in the Bruton agammaglobulinemia tyrosine kinase (BTK) gene are responsible for X-linked agammaglobulinemia (XLA). Unfolded or misfolded proteins can trigger stress pathways in the endoplasmic reticulum (ER), known as unfolded protein response (UPR). The aim was to clarify the involvement of UPR in XLA pathophysiology. By reverse transcription-quantitative PCR, we evaluated the expression of BTK and 12 UPR-related genes in eight patients. Moreover, we assessed the BTK protein expression and pattern in the patients' monocytes by flow cytometry and fluorescence immunocytochemistry. We found a reduced BTK expression in patients with stop codon mutations (P < 0.02). However, missense mutations did not affect BTK expression. Flow cytometry showed a reduction of BTK in patients which was corroborated by an absent or nonfunctional protein synthesis revealed by immunocytochemistry. In contrast with the other UPR-related genes, X-box binding protein 1 (XBP1) was markedly upregulated in the patients (P < 0.01), suggesting Toll-like receptor (TLR) activation since BTK directly interacts with TLRs as a negative regulator and XBP1 can be activated in direct response to TLR ligation. Different BTK mutations can be identified by the BTK expression. Inasmuch as UPR-related genes were downregulated or unaltered in patients, we speculate the involvement of the TLRs-XBP1 axis in the XLA pathophysiology. Such data could be the basis for further studies of this novel pathomechanism concerning XLA.3171-18

Assessing potential effects of a laccase extract over the enzymatichydrolysis of Eucalyptus bark residues

Lignocellulosic materials are rising as an alternative to petroleum, from which biofuels and numerous compounds may be produced. Eucalyptus barks, abundantly generated by pulp & paper mills, are a good example of such materials, being typically used for energy production. Holocellulose conversion of these materials is usually made by enzyme preparations, mainly acting on the hydrolysis of complex cellulose into monomer sugars. These materials, however, can still present a substantial amount of lignin, a well-documented enzymes barrier. This work aimed to assess how a laccases extract can influence the hydrolysis of eucalyptus bark and the best conditions for their action. Eucalyptus bark residues (EBR) were initially subjected to autohydrolysis with a severity (S0) of 3.84 [1]. The pre-treated solid was then hydrolyzed using Cellic CTec2, combined with a laccases-mediated treatment employing an extract prepared by the group of Maria de Lourdes Polizelli [2]. Potential effects of laccases were estimated through the quantification of the glucose produced over time and differences in the profile of enzymes adsorption onto the solid. The effects of laccases over the hydrolysis of EBR seemed to be dependent of numerous factors. For a solids load of 2 %, laccases addition simultaneously with cellulases had no positive effects but when added 24 h before cellulases, glucose production increased 11 %, possibly from an inferior electron donors competition with LPMOs on Cellic Ctec. Increasing laccases dosage from 2 to 10 IU/g solid led to a visible reduction of hydrolysis efficiency, suggesting possible toxicity/inhibition effects above a given level. Applying a washing step showed to be efficient in removing some of the formed phenolics, while its overall benefit seemed to depend on the extension of laccases action before being washed. When an efficient laccases treatment was conducted before the washing step, involving reduced mass transfer limitations and an adequate period of time, subsequent enzymatic hydrolysis produced nearly 30 % more glucose for a 8 % solids load. In accordance, there was also a significant increase on the levels of free Cel7A after hydrolysis of this new solid, suggesting important modifications on the levels and structure of its lignin. The utilization of laccases on the hydrolysis of lignocellulosic biomass may represent an interesting element for more efficient and economic processes.This work had the financial support from the Portuguese Foundation for Science and Technology under the scope of Project EcoTech (POCI-01-0145-FEDER-032206). The authors also acknowledge RAIZ for kindly providing the residues of eucalyptus bark.info:eu-repo/semantics/publishedVersio

Phase-shifted Bragg grating inscription in PMMA microstructured POF using 248 nm UV radiation

In this work we experimentally validate and characterize the first phase-shifted polymer optical fiber Bragg gratings (PS-POFBGs) produced using a single pulse from a 248 nm krypton fluoride laser. A single-mode poly (methyl methacrylate) optical fiber with a core doped with benzyl dimethyl ketal for photosensitivity improvement was used. A uniform phase mask customized for 850 nm grating inscription was used to inscribe these Bragg structures. The phase shift defect was created directly during the grating inscription process by placing a narrow blocking aperture in the center of the UV beam. The produced high-quality Bragg grating structures, presenting a double dips, reject 16.3 dB (97.6% reflectivity) and 13.2 dB (95.2% reflectivity) of the transmitted power, being therefore appropriate for sensing or other photonic applications. Its transmission spectrum possesses two sharp transmission notches, allowing a significant increase in measurement resolution compared to direct interrogation of a single grating. The reflection and transmission spectra when multiple phase shifts are introduced in the FBG structure are also shown. The PS-POFBG's strain, temperature, pressure, and humidity characteristics have been experimentally analyzed in detail to assess their potential usage as sensors