Non-invasive strategy in assessing asthma through biofluids metabolomics exploration: exhaled breath and urine potentialities

Asthma is a significant health issue in the pediatric population with a noteworthy growth over the years. The proposed challenge for this PhD thesis was the development of advanced methodologies to establish metabolomic patterns in urine and exhaled breath associated with asthma whose applicability was subsequently exploited to evaluate the disease state, the therapy adhesion and effect and for diagnostic purposes. The volatile composition of exhaled breath was studied combining headspace solid phase microextraction (HS-SPME) with gas chromatography coupled to mass spectrometry or with comprehensive two-dimensional gas chromatography coupled to mass spectrometry with a high resolution time of flight analyzer (GC×GC–ToFMS). These methodologies allowed the identification of several hundred compounds from different chemical families. Multivariate analysis (MVA) led to the conclusion that the metabolomic profile of asthma individuals is characterized by higher levels of compounds associated with lipid peroxidation, possibly linked to oxidative stress and inflammation (alkanes and aldehydes) known to play an important role in asthma. For future applications in clinical settings a set of nine compounds was defined and the clinical applicability was proven in monitoring the disease status and in the evaluation of the effect and / or adherence to therapy. The global volatile metabolome of urine was also explored using an HSSPME/GC×GC–ToFMS method and c.a. 200 compounds were identified. A targeted analysis was performed, with 78 compounds related with lipid peroxidation and consequently to oxidative stress levels and inflammation. The urinary non-volatile metabolomic pattern of asthma was established using proton nuclear magnetic resonance (1H NMR). This analysis allowed identifying central metabolic pathways such as oxidative stress, amino acid and lipid metabolism, gut microflora alterations, alterations in the tricarboxylic acid (TCA) cycle, histidine metabolism, lactic acidosis, and modification of free tyrosine residues after eosinophil stimulation. The obtained results allowed exploring and demonstrating the potential of analyzing the metabolomic profile of exhaled air and urine in asthma. Besides the successful development of analysis methodologies, it was possible to explore through exhaled air and urine biochemical pathways affected by asthma, observing complementarity between matrices, as well as, verify the clinical applicability.

Retinal nerve fiber layer analysis using GDx® in 49 patients with chronic phase DUSN

To describe retinal nerve fiber layer changes in late-stage diffuse unilateral subacute neuroretinitis eyes and compare these results with healthy eyes observed through nerve fiber analyzer (GDx®). Methods: This is a retrospective case-control study in which 49 eyes in late-stage diffuse unilateral subacute neuroretinitis were examined from May/97 to December/ 01. First, eyes with diffuse unilateral subacute neuroretinitis and healthy contralateral eyes (Control Group I) were statistically matched. Subsequently, eyes with diffuse unilateral subacute neuroretinitis were compared with eyes of healthy patients (Control Group II). Results: Eyes from Control Groups I and II had higher relative frequency of “within normal limits” status. Eyes from the diffuse unilateral subacute neuroretinitis (DUSN) Group had higher frequency of “outside normal limits” and “borderline” status. Control Groups I and II had absolute values different from the DUSN Group regarding all parameters (p<0.05), except for Symmetry in Control Groups I and II, Average thickness and Superior Integral in control group II. Conclusion: Patients with late-stage diffuse unilateral subacute neuroretinitis presented presumed decrease in nerve fiber layer thickness shown by GDx®. Retinal zones with larger vascular support and larger amount of nerve fibers presented higher decrease in the delay of the reflected light measured by the nerve fiber analyzer

Estudo morfológico da blenda polimérica poli(metacrilato de metila)/poli(tereftalato de etileno) reciclado (PMMA/PET)

Among the options for plastics modification more convenient, both from a technical-scientific and economic, is the development of polymer blends by processing in the molten state. This work was divide into two stages, with the aim to study the phase morphology of binary blend PMMA / PET blend and this compatibilized by the addition of the poly(methyl methacrylate-co-glycidyl methacrylate-co-ethyl acrylate) copolymer (MMA-GMA-EA). In the first stage is analyzed the morphology of the blend at a preliminary stage where we used the bottle-grade PET in a Haake torque rheometer and the effect of compatibilizer in this blend was evaluated. In the second stage the blend was processed using the recycled PET in a single screw extruder and subsequently injection molding in the shape of specimens for mechanical tests. In both stages we used a transmission electron microscopy (TEM) to observe the morphologies of the samples and an image analyzer to characterize them. In the second stage, as well as analysis by TEM, tensile test, scanning electron microscopy (SEM) and atomic force microscopy (AFM) was performed to correlate the morphology with the mechanical properties. The samples used in morphological analyzes were sliced by cryo-ultramicrotomy technique for the analysis by TEM and the analysis by SEM and AFM, we used the flat face of the block after cut cryogenic. It was found that the size of the dispersed phase decreased with the addition of MMA-GMA-EA in blends prepared in a Haake. In the tensile test, the values of maximum tensile strength and modulus of elasticity is maintained in a range between the value of pure PMMA the pure PET, while the elongation at break was influenced by the composition by weight of the PMMA mixture. The coupling agent corroborated the results presented in the blend PMMA / PETrec / MMA-GMA-EA (80/15/5 %w/w), obtained by TEM, AFM and SEM. It was concluded that the techniques used had a good morphologic correlation, and can be confirmed for final analysis of the morphological characteristics of the blends PMMA / PET

Análise das emissões gasosas de um veículo flex funcionando com diferentes combustíveis

Efforts in research and development of new technologies to reduce emission levels of pollutant gases in the atmosphere has intensified in the last decades. In this context, it can be highlighted the modern systems of electronic engine management, new automotive catalysts and the use of renewable fuels which contributes to reduce the environmental impact. The purpose of this study was a comparative analysis of gas emissions from a automotive vehicle, operating with different fuels: natural gas, AEHC or gasoline. To execute the experimental tests, a flex vehicle was installed on a chassis dynamometer equipped with a gas analyzer and other complementary accessories according to the standard guidelines of emission and security procedures. Tests were performed according to NBR 6601 and NBR 7024, which define the urban and road driving cycle, respectively. Besides the analysis of exhaust gases in the discharge tube, before and after the catalyst, using the suction probe of the gas analyzer to simulate the vehicle in urban and road traffic, were performed tests of fuel characterization. Final results were conclusive in indicating leaded gasoline as the fuel which most contributed with pollutant emissions in atmosphere and the usual gasoline being the fuel which less contributed with pollutant emissions in atmosphere

Avaliação da correlação entre os parâmetros, carbono orgânico total e elementos tóxicos, em solo de um posto de serviço e revenda de combustíveis

Today a major responsibility for the contamination of soil and groundwater and surface water are establishments known as gas stations of fuel which has attracted increasing attention from both the general population as the state agencies of environmental control due to leaks in storage tanks and mainly to disruption of pipe corrosion of tanks and pumping. Other services, like oil changes and car wash are also causes for concern in this type of establishment. These leaks can cause or waste produced, and the contamination of aquifers, serious health problems and public safety, since most of these stations located in urban areas. Based on this, the work was to evaluate soil contamination of a particular service station and fuel sales in the city of Natal, through the quantification of heavy metals like Cd, Cu, Cr, Ni, Pb, Zn of total organic carbon (TOC) and organic matter using different techniques such as optical emission spectrometry with inductively coupled plasma source (ICP OES), Total Organic Carbon analyzer and gravimetric analysis respectively. And also to characterize the soil through particle size analysis. Samples were taken in 21 georeferenced points and collected in the same period. The soils sampled in sampling stations P3, P5, P6, P10, P11, P12, P13, P14, P15, P17, P18 and P20 showed the smallest size fractions ranging from fine sand to medium sand. The other study sites ranged from fine sand to medium sand, except the point P8 showed that only the type size medium sand and P19, indicating a particle size of the coarse type. The small correlation of organic matter with the elements studied in this work suggests that these are not of anthropogenic origin but geochemical support

Obtenção de álcool alílico (PROP-2-EN-1-OL) a partir da glicerina derivada do biodiesel de óleo de mamona

In this work, biodiesel was produced from castor oil that was a byproduct glycerin. The molar ratio between oil and alcohol, as well as the use of (KOH) catalyst to provide the chemical reaction is based on literature. The best results were obtained using 1 mol of castor oil (260g) to 3 moles of methyl alcohol (138g), using 1.0% KOH as catalyst at a temperature of 260 ° C and shaken at 120 rpm. The oil used was commercially available, the process involves the reaction of transesterification of a vegetable oil with methyl alcohol. The product of this reaction is an ester, biodiesel being the main product and the glycerin by-product which has undergone treatment for use as raw material for the production of allyl alcohol. The great advantage of the use of glycerin to obtain allyl alcohol is that its use eliminates the large amount of waste of the biodiesel and various forms of insult to the environment. The reactions for the formation of allyl alcohol was conducted from formic acid and glycerin in a ratio 1/1, at a temperature of 260oC in a heater blanket, being sprayed by a spiral condenser for a period of 2 hours and the product obtained contains mostly the allylic alcohol .. The monitoring of reactions was performed by UV-Visible Spectrophotometer: FTIR Fourier transform, the analysis showed that these changes occur spectrometer indicating the formation of the product allylic alcohol (prop-2-en-1-ol) in the presence of water, This alcohol was appointed Alcohol GL. The absorption bands confirms that the reaction was observed in (υ C = C) 1470 -1600 cm -1 and (υ CO), 3610-3670 attributed to C = C groups and OH respectively. The thermal analysis was carried out in a thermogravimetric analyzer SDT Q600, where the mass and temperature are displayed against time, that allows checking the approximate rate of heating. The innovative methodology developed in the laboratory (LABTAM, UFRN), was able to treat the glycerine produced by transesterification of castor oil and used as raw material for production of allyl alcohol, with a yield of 80%, of alcohol, the same is of great importance in the manufacture of polymers, pharmaceuticals, organic compounds, herbicides, pesticides and other chemicals

Estudo de adulteração em méis brasileiros através de razão isotópica do carbono

Neste trabalho, objetivou-se analisar isotopicamente méis comercializados nas regiões Sul e Sudeste do Brasil, para a detecção de fraude. Foram colhidas amostras comerciais com registro no Serviço de Inspeção Federal, Estadual ou Municipal. As amostras foram submetidas à combustão no Analisador Elementar EA 1108 CHN e analisadas no espectrômetro de massas de razão isotópica DELTA-S (Finningan Mat). Os valores isotópicos (δ13C) dos méis in natura foram comparados aos de suas respectivas proteínas (padrão interno). Foram consideradas adulteradas as amostras cuja diferença entre o valor isotópico da proteína e do mel foi igual ou inferior a -1 . As amostras consideradas adulteradas pela análise isotópica foram submetidas a testes químicos qualitativos que não foram capazes de indicar adulteração para algumas delas. Das 61 amostras analisadas, 18,0% encontram-se adulteradas, sendo 11,5% na Região Sudeste e 6,5% na Região Sul. Ao contrário dos testes químicos, a análise isotópica mostrou-se eficaz em identificar e quantificar a adulteração de méis comerciais.

Emprego da citometria de fluxo na avaliação do perfil imunofenotípico de pacientes com leucemia linfocítica crônica

Chronic lymphocytic leukemia (B-CLL) is a clonal proliferation of mature B lymphocytes characterized by indolent clinical course. Biologically this clonallity is characterized by low expression of surface immunoglobulin (sIg) with restriction to a single immunoglobulin light chain associated with high expression of CD5 antigen and positivity to B cell antigens lymphocytes such as CD19, CD20 and CD23 and negativity to FMC7. The immunological profile and morphological analysis of lymphoid cells are the main means for the differential diagnosis of B-CLL from other chronic lymphoproliferative diseases. The aim of this study was to evaluate the expression pattern of a variety of membrane antigens in leukemic cells originating from patients with B-CLL. In this study, peripheral blood samples from 80 patients with B-CLL were analyzed by multiparametric flow cytometry in addition to routine hematologic exams, using a panel of monoclonal antibodies (MoAb): CD45/CD14, CD3/CD19/CD45, CD4/CD8 / CD3, CD20/CD5/CD3, CD3/CD16-56/CD45, CD2/CD7, FMC7/CD23, CD103/CD22/CD20, HLADR/CD38, CD10/CD19, CD1a, CD11b and also IgM/gD, kappa and lambda immunoglobulin light chains for the detection of surface immunoglobulin and clonal restriction for immunoglobulin light chain. The Hematological data were obtained from the hematological analyzer and cytomorphological analysis in blood film stained by Leishmann. The study samples consisted of 45 men and 35 women, ages ranging from 55 to 84 years (mean 65 years). Complete white blood count showed count ranging from 10.0 to 42.0 x 109/l. (mean 50.0 x 109/l) and lymphocytes count greater than 5.0 x 109/l in all cases. The neoplastic cells displayed B-CLL phenotype (CD5+/CD19+/CD20+/HLADR+/CD23+) in the vast majority of the cases, associated to failed to stain for T cell markers (CD1a, CD2, CD4, CD3, CD7, CD8), CD103, CD14 and FMC7. Leukemic cells of most patients also expressed low intensity of IgM and IgD with restricted kappa light chain, in most cases (59,7%). This observation highlights the importance of immunophenotyping for correct diagnosis of chronic lymphoproliferative syndromes and the panel of MoAb used was sufficient for diagnostic confirmation of B-CLL

Soroprevalência da infecção pelo Trypanosoma cruzi no município de Caicó, RN, e caracterização genética por RAPD do parasito isolado de humanos e de triatomíneos no semiárido potiguar

Trypanosoma cruzi infection was evaluated in 390 resident individuals in different rural communities of Caicó municipality, State of Rio Grande do Norte (RN). Of 28 investigated communities the soroprevalence of T. cruzi infection was 2.8% in eight rural communities individuals. The epidemiological characteristics of seropositive shown that the age ranged from 22 to 64 years, being significantly raised from 31 years (90.9%). The female gender was predominant and low education degree. Those individuals reported that they never donated blood, but they had direct contact with triatomines bug. The isolation of the parasite was performed by blood culture and xenoculture methods to determine the genetic variability of the samples. Twenty seven T. cruzi isolates were analyzed by RAPD as genetic marker using three random primers (M13-40, gt11-F and L15996). The T. cruzi isolates showed 73.7% of shared bands considering the average obtained with the three primers, and were genetically well correlated. Using this marker it was possible to separate the populations of the parasite in three distinct groups. The first group composed by isolates obtained of triatomines and humans from four different districts (Caicó, Caraúbas, Serra Negra doNorte and Governador Dix-Sept Rosado); the second contained isolates obtained of triatomines of two different species (T. brasiliensis and P. lutzi) captured in Caraúbas and Serra Negra do Norte. The third grouped isolates obtained from humans of Angicos and Caicó municipalities. In different localities of distinct mesoregions, State of RN, a profile genetic well correlated was identified among all isolates and the presence of three distinct groups of the parasite circulating among vertebrate and invertebrate hosts

(-)-Hinokinin-loaded poly(d,l-lactide-co-glycolide) microparticles for Chagas disease

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Funções de pedotransferência para a curva de resistência do solo à penetração

Funções de pedotransferência são regressões utilizadas para estimar atributos edáficos dependentes a partir de atributos independentes e de fácil determinação. Nesse sentido, são propostas na literatura diversas funções de pedotransferência que visam predizer a resistência do solo à penetração. Objetivou-se, portanto, com este trabalho, desenvolver e comparar a eficiência de cinco funções de pedotransferência para a curva de resistência do solo à penetração, presentes na literatura, por meio do ajuste de dados obtidos tanto com o penetrômetro de impacto (campo) quanto com o penetrômetro eletrônico (laboratório), em um Latossolo manejado sob diferentes modos (convencional e plantio direto). Foram coletadas amostras indeformadas de solo na entrelinha das culturas, nas camadas de 0-0,10, 0,10-0,20 e 0,20-0,30 m, logo após a semeadura, no florescimento e na colheita, para determinação dos atributos físico-hídricos do solo e também da resistência do solo à penetração, com o uso do penetrômetro eletrônico. A resistência do solo à penetração, obtida com o penetrômetro de impacto, foi determinada conforme a variação do conteúdo de água no solo ao longo do ciclo das culturas. As curvas ajustadas de resistência do solo à penetração tiveram a precisão e a acurácia testadas por meio de parâmetros estatísticos e foram comparadas pelo teste F. Houve sobreposição dos valores estimados pelo ajuste das curvas, evidenciando que a maneira de obtenção da resistência do solo à penetração (campo ou laboratório) não influenciou a relação entre a resistência à penetração e os atributos do solo. As equações RP = aUg b; RP = a(1-Ug)b; RP = ae bUg e RP = a + be não diferiram e foram as mais precisas e acuradas na predição da resistência do solo à penetração.

Alterações morfométricas no plexo mioentérico do cólon menor equino distendido experimentalmente

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Modelo de avaliação de risco : São o crescimento e a estequiometria dos girinos afetados pelo efeito interativo entre a presença do predador e da densidade de coespecíficos?

Many prey organisms change their phenotype to reduce the predation risk. However, such changes are associated with trade-offs, and can have negative effects on prey growth or reproduction. Understand how preys assess the predation risk is essential to evaluate the adaptive value of predator-induced phenotypic and its ecological consequences. In this study, we performed a mesocosm experiment to test: i) if growth and stoichiometry of Lithobates catesbeianus tadpoles is altered in response to giant water bug presence (Belostoma spp.); ii) if these responses depend on tadpoles density in environment. Here, we show that tadpoles growth and stoichiometry are not changed by predator presence, neither in low nor in high densities. Our results suggest that tadpoles exposed to predation risk regulate their physiology to preserve the elemental stoichiometric homeostase of their body and excretion. Further, point out to need for future studies that elucidate under what conditions growth and stoichiometry are changed in response to predation risk

Virulência de Aphanocladium album (Preuss) Gams e Verticillium lecanii (Zimm.) Viégas (Deuteromycotina: Hyphomycetes) para o Percevejo-de-renda da seringueira, Leptopharsa heveae (Drake & Poor) (Hemiptera: Tingidae)

The lace bug, Leptopharsa heveae is an insect that causes serious damage on rubber trees. In laboratory condition, the strains of Verticillium lecanii (ARSEF 6430, 6431 e 6432) and Aphanocladium album (ARSEF 6433) were tested on third-and fifth-instar nymphs and adults of L. heveae to evaluate their virulence using 2.4 x 10(5) and 2.4 x 10(7) conidia/mL. The bioassays were carried out using Petri dishes whose inner bottoms were covered with damp filter papers. Each Petri dish contained five insects and one rubber tree leaflet. The plates were covered with PVC film to provide high relative humidity, maintained at 26 +/- 0.5 degrees C and a photophase of 14 hours. The Probit analysis was calculated from mortality date of insects killed by fungi. In the highest concentrations, ARSEF 6430 was more virulent for third instar nymphs, and the LT50 was 1.9 days. For the fifth instar, the strains ARSEF 6430, 6433 and 6432 showed similar virulence with LT50 of 2.6, 2.6 and 3.2 days, respectively. For adults, ARSEF 6431 was the most virulent strain with the LT50 recorded at 2.0 days. The smallest concentration did not always cause more than 50% mortality.

Biologia de Leptopharsa heveae Drake & Poor (Heteroptera: Tingidae) e a relação de suas exigências térmicas com a flutuação populacional em seringueira

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)