Investigation of urinary volatile organic metabolites as potential cancer biomarkers by solid-phase microextraction in combination with gas chromatography-mass spectrometry

BACKGROUND: Non-invasive diagnostic strategies aimed at identifying biomarkers of cancer are of great interest for early cancer detection. Urine is potentially a rich source of volatile organic metabolites (VOMs) that can be used as potential cancer biomarkers. Our aim was to develop a generally reliable, rapid, sensitive, and robust analytical method for screening large numbers of urine samples, resulting in a broad spectrum of native VOMs, as a tool to evaluate the potential of these metabolites in the early diagnosis of cancer. METHODS: To investigate urinary volatile metabolites as potential cancer biomarkers, urine samples from 33 cancer patients (oncological group: 14 leukaemia, 12 colorectal and 7 lymphoma) and 21 healthy (control group, cancer-free) individuals were qualitatively and quantitatively analysed. Dynamic solid-phase microextraction in headspace mode (dHS-SPME) using a carboxenpolydimethylsiloxane (CAR/PDMS) sorbent in combination with GC-qMS-based metabolomics was applied to isolate and identify the volatile metabolites. This method provides a potential non-invasive method for early cancer diagnosis as a first approach. To fulfil this objective, three important dHS-SPME experimental parameters that influence extraction efficiency (fibre coating, extraction time and temperature of sampling) were optimised using a univariate optimisation design. The highest extraction efficiency was obtained when sampling was performed at 501C for 60min using samples with high ionic strengths (17% sodium chloride, wv 1) and under agitation. RESULTS: A total of 82 volatile metabolites belonging to distinct chemical classes were identified in the control and oncological groups. Benzene derivatives, terpenoids and phenols were the most common classes for the oncological group, whereas ketones and sulphur compounds were the main classes that were isolated from the urine headspace of healthy subjects. The results demonstrate that compound concentrations were dramatically different between cancer patients and healthy volunteers. The positive rates of 16 patients among the 82 identified were found to be statistically different (Po0.05). A significant increase in the peak area of 2-methyl3-phenyl-2-propenal, p-cymene, anisole, 4-methyl-phenol and 1,2-dihydro-1,1,6-trimethyl-naphthalene in cancer patients was observed. On average, statistically significant lower abundances of dimethyl disulphide were found in cancer patients. CONCLUSIONS: Gas chromatographic peak areas were submitted to multivariate analysis (principal component analysis and supervised linear discriminant analysis) to visualise clusters within cases and to detect the volatile metabolites that are able to differentiate cancer patients from healthy individuals. Very good discrimination within cancer groups and between cancer and control groups was achieved.

Effectiveness of high-throughput miniaturized sorbent- and solid phase microextraction techniques combined with gas chromatography–mass spectrometry analysis for a rapid screening of volatile and semi-volatile composition of wines: a comparative study

In this study the feasibility of different extraction procedures was evaluated in order to test their potential for the extraction of the volatile (VOCs) and semi-volatile constituents (SVOCs) from wines. In this sense, and before they could be analysed by gas chromatography–quadrupole first stage masss spectrometry (GC–qMS), three different high-throughput miniaturized (ad)sorptive extraction techniques, based on solid phase extraction (SPE), microextraction by packed sorbents (MEPS) and solid phase microextraction (SPME), were studied for the first time together, for the extraction step. To achieve the most complete volatile and semi-volatile signature, distinct SPE (LiChrolut EN, Poropak Q, Styrene-Divinylbenzene and Amberlite XAD-2) and MEPS (C2, C8, C18, Silica and M1 (mixed C8-SCX)) sorbent materials, and different SPME fibre coatings (PA, PDMS, PEG, DVB/CAR/PDMS, PDMS/DVB, and CAR/PDMS), were tested and compared. All the extraction techniques were followed by GC–qMS analysis, which allowed the identification of up to 103 VOCs and SVOCs, distributed by distinct chemical families: higher alcohols, esters, fatty acids, carbonyl compounds and furan compounds. Mass spectra, standard compounds and retention index were used for identification purposes. SPE technique, using LiChrolut EN as sorbent (SPELiChrolut EN), was the most efficient method allowing for the identification of 78 VOCs and SVOCs, 63 and 19 more than MEPS and SPME techniques, respectively. In MEPS technique the best results in terms of number of extractable/identified compounds and total peak areas of volatile and semi-volatile fraction, were obtained by using C8 resin whereas DVB/CAR/PDMS was revealed the most efficient SPME coating to extract VOCs and SVOCs from Bual wine. Diethyl malate (18.8 ± 3.2%) was the main component found in wine SPELiChrolut EN extracts followed by ethyl succinate (13.5 ± 5.3%), 3-methyl-1-butanol (13.2 ± 1.7%), and 2-phenylethanol (11.2 ± 9.9%), while in SPMEDVB/CAR/PDMS technique 3-methyl-1-butanol (43.3 ± 0.6%) followed by diethyl succinate (18.9 ± 1.6%), and 2-furfural (10.4 ± 0.4%), are the major compounds. The major VOCs and SVOCs isolated by MEPSC8 were 3-methyl-1-butanol (26.8 ± 0.6%, from wine total volatile fraction), diethyl succinate (24.9 ± 0.8%), and diethyl malate (16.3 ± 0.9%). Regardless of the extraction technique, the highest extraction efficiency corresponds to esters and higher alcohols and the lowest to fatty acids. Despite some drawbacks associated with the SPE procedure such as the use of organic solvents, the time-consuming and tedious sampling procedure, it was observed that SPELiChrolut EN, revealed to be the most effective technique allowing the extraction of a higher number of compounds (78) rather than the other extraction techniques studied.

Characterization of volatile substances in apples from rosaceae family by headspace solid‐phase microextraction followed by GC‐qMS

The volatile composition of different apple varieties of Malus domestica Borkh. species from different geographic regions at Madeira Islands, namely Ponta do Pargo (PP), Porto Santo (PS), and Santo da Serra (SS) was established by headspace solid-phase microextraction (HS-SPME) procedure followed by GC-MS (GC-qMS) analysis. Significant parameters affecting sorption process such as fiber coating, extraction temperature,extractiontime,sampleamount,dilutionfactor,ionicstrength,anddesorption time,wereoptimizedanddiscussed.TheSPMEfibercoatedwith50/30 lmdivinylbenzene/carboxen/PDMS (DVB/CAR/PDMS) afforded highest extraction efficiency of volatile compounds, providing the best sensitivity for the target volatiles, particularly whenthesampleswereextractedat508Cfor30 minwithconstantmagneticstirring. A qualitative and semi-quantitative analysis between the investigated apple species has been established. It was possible to identify about 100 of volatile compounds amongpulp(46,45,and39),peel(64,60,and64),andentirefruit(65,43,and50)inPP, PS,andSSapples,respectively.Ethylesters,terpenes,andhigheralcoholswerefound tobethemostrepresentativevolatiles. a-Farnesene,hexan-1-olandhexyl2-methylbutyratewerethecompoundsfoundinthevolatileprofileofstudiedappleswiththelargestGCarea,representing,onaverage,24.71,14.06,and10.80%ofthetotalvolatilefractionfromPP,PS,andSSapples.InPPentireapple,themostabundantcompoundsidentified were a-farnesene (30.49%), the unknown compound m/z (69, 101, 157) (21.82%) andhexylacetate(6.57%).RegardingPSentireapplethemajorcompoundswere a-farnesene(16.87%),estragole(15.43%),hexan-1-ol(10.94),andE-2-hexenal(10.67).a-Farnesene(30.3%),hexan-1-ol(18.90%),2-methylbutanoicacid(4.7%),andpentan-1-ol(4.6%) werealsofoundasSSentireapplevolatilespresentinahigherrelativecontent.Principal component analysis (PCA) of the results clustered the apples into three groups according to geographic origin. Linear discriminant analysis (LDA) was performed in order to detect the volatile compounds able to differentiate the three kinds of apples investigated. The most important contributions to the differentiation of the PP, PS, and SS apples were ethyl hexanoate, hexyl 2-methylbutyrate, E,E-2,4-heptadienal, pethylstyrene,andE-2-hexenal.

A gestão da informação sob a luz do Enterprise Content Management (ECM): um estudo de caso em uma universidade pública

This work involves the organization and content perspectives on Enterprise Content Management (ECM) framework. The case study at the Federal University of Rio Grande do Norte was based on ECM model to analyse the information management provided by the three main administrative systems: The Integrated Management of Academic Activities (SIGAA), Integrated System of Inheritance, and Contracts Administration (SIPAC) and the Integrated System for Administration and Human Resources (SIGRH). A case study protocol was designed to provide greater reliability to research process. Four propositions were examined in order to reach the specific objectives of identification and evaluation of ECM components from UFRN perspective. The preliminary phase provided the guidelines for the data collection. In total, 75 individuals were interviewed. Interviews with four managers directly involved on systems design were recorded (average duration of 90 minutes). The 70 remaining individuals were approached in random way in UFRN s units, including teachers, administrative-technical employees and students. The results showed the presence of many ECM elements in the management of UFRN administrative information. The technological component with higher presence was "management of web content / collaboration". But initiatives of other components (e.g. email and document management) were found and are in continuous improvement. The assessment made use of eQual 4.0 to examine the effectiveness of applications under three factors: usability, quality of information and offered service. In general, the quality offered by the systems was very good and walk side by side with the obtained benefits of ECM strategy adoption in the context of the whole institution

Integrated sediment quality assessment in Paranagua Estuarine System, Southern Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Yeast extract and prebiotic in pre-initial phase diet for broiler chickens raised under different temperatures

Objetivou-se nesta pesquisa avaliar o desempenho, o rendimento de carcaça e a morfometria intestinal de frangos de corte criados em diferentes temperaturas e que receberam na fase pré-inicial ração contendo ou não extrato de leveduras e prebiótico. Foram utilizados 1.440 pintos machos de 1 dia de idade, criados em diferentes câmaras climáticas. As rações, acrescidas ou não de extrato de leveduras e prebiótico, foram oferecidas somente na fase pré-inicial (1 a 7 dias). A partir do oitavo dia, todas as aves receberam a mesma ração, reajustada de acordo com as recomendações usuais. Adotou-se o delineamento experimental inteiramente casualizado em arranjo fatorial 3 × 2 × 2, composto de três temperaturas de criação (calor, conforto e frio) e dois níveis de extrato de leveduras (com ou sem) e prebiótico (com ou sem). O desempenho das aves foi avaliado considerando o ganho de peso, o consumo de ração, a conversão alimentar e a viabilidade aos 42 dias de idade. Também foram avaliados o rendimento de carcaça e a morfometria intestinal. O calor ambiente prejudicou o desempenho e o rendimento de carcaça. A inclusão de prebiótico na ração pré-inicial aumentou o ganho de peso e melhorou a conversão alimentar das aves criadas no calor. A inclusão dos produtos na ração de frangos de corte criados em ambiente de calor e no frio tem efeito benéfico sobre as vilosidades das aves.

Transcriptomics and systems biology analysis in identification of specific pathways involved in cacao resistance and susceptibility to witches' broom disease

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Comparison of three protocols for superovulation of brown brocket deer (Mazama gouazoubira)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Digging effort in leaf-cutting ant queens (Atta sexdens rubropilosa) and its effects on survival and colony growth during the claustral phase

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

Knowledge-based integrated production management model

Purpose - This paper aims to propose a model of production management that integrates knowledge management, as a third dimension, to the production and work dimensions and to identify factors that promote a favorable context for knowledge sharing and results achievement in the production operations shop floor environment.Design/methodology/approach - The model proposed is built from opportunities identified in the literature review.Findings - The factors in the model integrate its three main components: knowledge management, production organization and work organization, providing a representation of the dynamics of the workplace and shop floor environment.Practical implications - The proposed model and its factors allow managers to better understand and to improve the organization activities, because it integrates knowledge management with the production organization and work organization components of traditional models.Originality/value - Literature acknowledges the role of knowledge as competitive advantage, but it is still dealt in an implicit way within the traditional models of production management. This paper proposes a model and factors that provide a favorable context for tacit knowledge sharing and results achievement in the production operations shop floor environment. The model explicitly integrates knowledge management with traditional models' components.

Mathematical modeling of the ester oil-refrigerant R134A mixture two-phase flow with foam formation through a small diameter tube

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Modelagem do clima de ondas e seus efeitos sobre as feições morfológicas costeiras no litoral setentrional do Rio Grande do Norte

This thesis presents the results of application of SWAN Simulating WAves Nearshore numerical model, OF third generation, which simulates the propagation and dissipation of energy from sea waves, on the north continental shelf at Rio Grande do Norte, to determine the wave climate, calibrate and validate the model, and assess their potential and limitations for the region of interest. After validation of the wave climate, the results were integrated with information from the submarine relief, and plant morphology of beaches and barrier islands systems. On the second phase, the objective was to analyze the evolution of the wave and its interaction with the shallow seabed, from three transverse profiles orientation from N to S, distributed according to the parallel longitudinal, X = 774000-W, 783000-W e 800000-W. Subsequently, it was were extracted the values of directional waves and winds through all the months between november 2010 to november 2012, to analyze the impact of these forces on the movement area, and then understand the behavior of the morphological variations according to temporal year variability. Based on the results of modeling and its integration with correlated data, and planimetric variations of Soledade and Minhoto beach systems and Ponta do Tubarão and Barra do Fernandes barrier islands systems, it was obtained the following conclusions: SWAN could reproduce and determine the wave climate on the north continental shelf at RN, the results show a similar trend for the measurements of temporal variations of significant height (HS, m) and the mean wave period (Tmed, s); however, the results of parametric statistics were low for the estimates of the maximum values in most of the analyzed periods compared data of PT 1 and PT 2 (measurement points), with alternation of significant wave heights, at times overrated with occasional overlap of swell episodes. By analyzing the spatial distribution of the wave climate and its interaction with the underwater compartmentalization, it was concluded that there is interaction of wave propagation with the seafloor, showing change in significant heights whenever it interacts with the seafloor features (beachrocks, symmetric and asymmetric longitudinal dunes, paleochannel, among others) in the regions of outer, middle and inner shelf. And finally, it is concluded that the study of the stability areas allows identifications of the most unstable regions, confirming that the greatest range of variation indicates greater instability and consequent sensitivity to hydrodynamic processes operating in the coastal region, with positive or negative variation, especially at Ponta do Tubarão and Barra do Fernandes barrier islands systems, where they are more susceptible to waves impacts, as evidenced in retreat of the shoreline

Contribuições para a detecção e identificação de faltas em inversores Multiníveis

This work presents contributions in the detection and identication of faults in multilevel inverters through the study of the converters behavior under these operation conditions. Basically, the approached fault consists of an open-circuit in any switch of a three-level clamped diode inverter. The converter operation is characterized in the pre and post-fault states. A wave form behavior analysis of the pole voltage, phase current and dc-bus current is also done, which highlights characteristics that allow the detection of failure and, even, under favorable conditions, the identication of the faulty device. A compensation strategy of the approached fault (open-switch) is also investigated with the purpose of maintaining the driving system operational when a failure occurs. The proposed topology uses SCRs in parallel with the internal switches of the inverter, which allows, in some occasions, the full utilization of the dc-bus

Cytokines and acute-phase serum proteins as markers of inflammatory regression during pulmonary tuberculosis treatment

Tuberculosis is still increasing and was declared a worldwide sanitary emergency by the World Health Organization (WHO) in 1995. Its control is difficult due to long treatment duration and lack of markers of treatment success or failure. Cytokines such as IFN-gamma and TNF-alpha, a central factor in immune response against Mycobacterium tuberculosis, are responsible for the interaction between T lymphocytes and the infected macrophage and are also produced during this interaction. As proinflammatory cytokines have a close relationship with mycobacteria clearance, in fact even preceding it, they could be used as markers for inflammatory activity and response to treatment. Proinflammatory cytokines act in the liver and stimulate a strong local and systemic acute-phase response as a result of homeostatic and physiological responses also induced by them. Acute-phase proteins produced by cytokine activity are useful diagnostic markers that could also be used to monitor treatment response as they can be serially quantified. The objective of this study was to evaluate IFN-gamma, TNF-alpha, IL-10 and TGF-beta production in supernatant of peripheral blood mononuclear cell (PBMC) and monocyte (MO) cultures, as well as serum acute-phase response through total protein, albumin, globulin, C-reactive protein (CRP), alpha-1-acid glycoprotein (AGP), and erythrocyte sedimentation rate (ESR) as regression markers of inflammatory response during pulmonary tuberculosis treatment. Twenty blood donors (G1) from the Blood Bank at Botucatu School of Medicine's University Hospital (BSM-UH) were evaluated once and 28 pulmonary tuberculosis patients (G2): 13 from BSM-UH and 15 from the Bauru State Health Secretariat. Patients were evaluated at three moments of treatment: before (M1), at three months (M2), and at the end (M3). Cytokines were determined in 20ml of peripheral blood (ELISA), with or without activation: lipopolysaccharide (LPS) for MO culture and phytohemagglutinin (PHA) for PBMC culture. Acute-phase protein behavior in G2 throughout treatment was: Globulins: M1> M2, M1> M3 (rho < 0.001); CRP: M1> M2> M3 (.< 0.001); AGP for men: M1> M2, M1> M3 (rho < 0.001); ESR for men: M1> M2, M1> M3 (rho < 0.0016) and for women: M1> M2 (.< 0.025). Comparison between cytokine levels found in supernatant of MO and PBMC cultures, with and without stimulus, in G1 and G2 during treatment showed: TNF-alpha (with/ without LPS) at M1: G2> G1; at M2: G2> G1 (rho < 0.001); (without LPS) at M3: G2> G1 (rho < 0.001), (with LPS) at M3: G2> G1 (rho < 0.028); IFN-. (with and without PHA) at M1: G2> G1; at M2: G2> G1 (rho < 0.001); IL-10 (with and without LPS) at M1: G2> G1; at M2: G2> G1; at M3: G2> G1 (rho < 0.001); TGF-beta (with and without LPS) at M1: G2> G1; at M2: G2> G1 (rho < 0.001), (without LPS) at M3: G2> G1 (rho < 0.001). In G2, all cytokines in supernatant of MO and PBMC cultures, with and without stimulus, showed: M1> M2> M3 (rho < 0.01). Levels of globulins, CRP, AGP, and ESR in patients with pulmonary tuberculosis before treatment (M1) were significantly higher than reference values, suggesting their use as diagnostic markers and indicators of treatment. The CRP decreasing values along treatment could be taken as a marker of the regression of inflammatory process and of response to treatment in patients with pulmonary tuberculosis.Regarding cytokines, there was significant increase in TNF-alpha, IFN-gamma, IL-10, and TGF-alpha levels before and at three months treatment, with and without stimulus; in TNF-a and IL-10 lvels, with and without stimulus, as well as in TGF-alpha levels without stimulus at six months. Patients had higher levels of all studied cytokines than controls before treatment, and these values decreased along treatment. In this study, pulmonary tuberculosis patients showed a Th0 cytokine profile before treatment, with the production of both Th1 (IFN-gamma) and Th2 (IL-10) cytokines, in addition to TNF-alpha inflammatory and TGF-alpha regulatory and fibrosis-inducer cytokines. At the end of treatment, all had evolved to Th2 profile, probably in an attempt to reduce the harmful effects of the proinflammatory activity of the Th1 cytokine profile and of the still above-normal levels of TNF-alpha. The high levels of TGF-alpha, also found in these patients, are related to its important role in the extracellular matrix deposition and fibrosis induction that characterize tuberculosis healing process. IFN-gamma was the only cytokine reaching normal levels at the end of treatment, which suggests its use as a marker of response to treatment.

As relações entre tectonismo e seqüências deposiconais no rifte potiguar - porção SW do Graben de Umbuzeiro, bacia potiguar emersa

Through an integrated approach, using litho, chrono and biostratigraphic data, the relative importance of climate variations and tectonics were recognized in rift sediments of the onshore Potiguar Basin, Northeast Brazil. Concepts of sequence stratigraphy were applied as a template to integrate sedimentological and geochemical data (oxygen isotopes), as well as quantitative palynologic methods to address and recognize the main depositional patterns produced in a rift basin. The main objective was to address the relative importance of climate changes and tectonics to the resultant stratigraphic architecture. The results of computer simulations of sedimentary basin fills of rift basins were quite useful to test working hypothesis and mimic the process of filling a half graben during a rift event. The studied section includes a neovalanginian-eobarremian (Lower Cretaceous) rift interval from the Pendência Formation, located in the southwestern portion of Umbuzeiro Graben, in the offshore Potiguar Basin. The depositional setting is interpreted as progradational deltaic system entering a lake from its flexural margin. Sismoestratigraphyc and well logs analyses allowed to interpret two regressive intervals (Green and Yellow Sequences), separated by a broad transgressive interval (Orange Sequence), known as the Livramento Shale. The depositional history encompass three stages: two tectonically active phases, during the deposition of the Green and Yellow Sequences, and a tectonically quiescent phase, during the deposition of the Orange Sequence. Paleoclimatic interpretation, based on quantitative palynology and geochemical data (��18O), suggests a tendency to arid conditions during the tectonically active phases and wet conditions during the tectonically quiescent phase. Stratigraphic modeling and backstripping techniques, supported by paleoclimatic/paleoecologic interpretations provide a powerful methodology to evaluate the tectonic and climatic controls on tectonic lakes