Objectives: Intact cell peptidome profiling (ICPP) with MALDI-ToF Mass-Spectrometry holds promise as a non invasive method to detect head and neck squamous cell carcinoma (HNSCC) objectively, which may improve the early diagnosis of oral cancer tremendously. The present study was designed to discriminate between tumour samples and non-cancer controls (healthy mucosa and oral lesions) by analysing complete spectral patterns of intact cells using MALDI-ToF MS. Material and Methods: In the first step, a data base consisting of 26 patients suffering from HNSCC was established by taking brush biopsy samples of the diseased area and of the healthy buccal mucosa of the respective contralateral area. After performing MALDI-ToF MS on these samples, classification analysis was used as a basis for further classification of the blind study composed of additional 26 samples including HNSCC, oral lesions and healthy mucosa. Results: By analyzing spectral patterns of the blind study, all cancerous lesions were defined accurately. One incorrect evaluation (false positive) occurred in the lesion cohort, leading to a sensitivity of 100%, a specificity of 93% and an overall accuracy of 96.5%. Conclusion: ICPP using MALDI-ToF MS is able to distinguish between healthy and cancerous mucosa and between oral lesions and oral cancer with excellent sensitivity and specificity, which may lead to a more impartial early diagnosis of HNSCC.

The aim of the study was to evaluate the total viable count and detects each microorganism by the use of mass spectroscopy. In the period of June to December 2017 35 samples of pasteurized milk from two vending machines (Hodkovice, Suchdol) were tested. For comparison samples were divided to warmer season (June-Septemeber) and colder season (October-December) sampling. The total amount of microorganism was determined in each sample of milk and total viable count was estimated by the use of mass spectroscopy MALDI-TOF. The total viable count in the tested pasteurized milk were from 3,27 log KTJ/ml up to 7,35 log KTJ/ml. The mass spectroscopy detected 210 microorganisms belonging to 20 families of gram-positive and gram-negative bacteria and one species of eukaryot organism. The most often identified species belong the Enterobacteriaceae family.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-05-10T22:31:41Z No. of bitstreams: 1 MariaCarolinaSoaresLopes_DISSERT.pdf: 2639801 bytes, checksum: ec2e4bc4eef612ece59848a3c3979ea1 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-05-17T20:06:54Z (GMT) No. of bitstreams: 1 MariaCarolinaSoaresLopes_DISSERT.pdf: 2639801 bytes, checksum: ec2e4bc4eef612ece59848a3c3979ea1 (MD5) / Made available in DSpace on 2016-05-17T20:06:54Z (GMT). No. of bitstreams: 1 MariaCarolinaSoaresLopes_DISSERT.pdf: 2639801 bytes, checksum: ec2e4bc4eef612ece59848a3c3979ea1 (MD5) Previous issue date: 2015-05-14 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / As Infec??es Hospitalares (IH) se constituem na principal causa de morbidade e mortalidade em pacientes hospitalizados. Relatos de isolamentos de Acinetobacter multirresistentes a partir de esp?cimes cl?nicos obtidos de pacientes internados bem como do ambiente hospitalar s?o cada vez mais frequentes. Assim, esse projeto tem como objetivo caracterizar fenotipica e molecularmente, isolados de Acinetobacter spp. quanto ? susceptibilidade aos antimicrobianos. Os isolados de Acinetobacter foram coletados em quatro hospitais localizados na cidade do Natal-RN, no per?odo de 2013 a 2014. A identifica??o dos isolados foi realizada atrav?s de provas laboratoriais convencionais, atrav?s do sistema MALDI-TOF e atrav?s da pesquisa do gene blaOXA51. A susceptibilidade aos antimicrobianos foi avaliada pela metodologia de disco-difus?o. Para a droga tigeciclina, a concentra??o inibit?ria m?nima (CIM) foi determinada atrav?s do E-test . Al?m disso, foram realizados testes de triagem para as enzimas AmpC, ESBL e Carbapanemases e pesquisa dos genes para as carbapenemases (IMP-1, VIM-1, NDM-1, KPC-2, OXA-23,OXA-24,OXA-58,OXA-143), atrav?s da t?cnica da Rea??o em Cadeia da Polimerase (PCR). N?o houve 100% de concord?ncia entre os resultados obtidos pelas t?cnicas utilizadas para a identifica??o dos isolados. 90% e 81,8% das amostras foram identificadas como A.baumannii pelos testes convencionais e MALDI-TOF, respectivamente e a positividade do blaOXA51 aconteceu em 97,1% das amostras . No ambiente hospitalar, os locais mais contaminados por essa esp?cie foi a bancada de procedimentos e o piso. A maioria (58,2%) das cepas de Acinetobacter apresentaram resist?ncia ? tr?s ou mais classes de antibi?ticos. A positividade no teste para detec??o de AmpC foi de 5,8%, no Teste de Hodge foi de 61,1% e para o teste de detec??o de Metallo-?-lactamase foi de 78,5%. Nenhuma amostra se mostrou produtora de ESBL. Os genes mais encontrados, al?m do blaOXA51 foram o blaOXA23 e blaOXA143. A m?dia da preval?ncia de Acinetobacter nos hospitais estudados foi de 7,6% em amostra cl?nicas e de 12,8% nas amostras do ambiente hospitalar, a sazonalidade foi demonstrada e v?rios fatores relacionados com a multirresist?ncia foram estatisticamente relevantes. O elevado n?mero de Acinetobacter multirresistente aos antimicrobianos, isolados a partir de pacientes e sobretudo das superf?cies inanimadas dos hospitais ? preocupante. Esse cen?rio pode comprometer tanto os tratamentos emp?ricos de pacientes em estado grave quanto ?s estrat?gias de controle de infec??o hospitalar. / The Hospital Infections are the main cause of morbidity and mortality in hospitalized patients. Isolations reports of multidrug-resistant Acinetobacter from clinical specimens obtained from hospitalized patients and the hospital are more frequent. Thus, this study aimed to characterize molecular and phenotypic isolated from Acinetobacter sp. for susceptibility to antimicrobials, with emphasis on ?-lactams. Clinical isolates and hospital surfaces isolates of Acinetobacter sp. were collected in four hospitals located in the city of Natal-RN, from March 2013 to March 2014. The identification of the isolates was carried out by standard laboratory tests, the MALDI-TOF system and research blaOXA-51 gene. The antimicrobial susceptibility was evaluated by disk diffusion method. For tigecycline drug, the minimum inhibitory concentration (MIC) was determined by the E-test. Furthermore, screening tests were performed for AmpC enzymes, and ESBL Carbapanemases and research of genes for carbapenemases (IMP-1, VIM-1, NDM-1, KPC-2, OXA-23, OXA-24, OXA- 58, OXA-143) by technique of polymerase chain reaction (PCR). Factors associated with the presence of multidrug resistance were also studied. 242 samples were studied, with 155 clinical samples and 87 samples of hospital surfaces. The mean occurrence of Acinetobacter sp. in the hospitals studied was 7.6% in clinical sample and 12.8% in samples of hospital surfaces. In this, the sites most contaminated by this bacterium was the bench procedures and the floor. Two hundred and thirty seven samples (97.9%) and 198 (81.8%) of the samples were identified as A.baumannii by conventional tests and MALDI-TOF, respectively, and the positivity of blaOXA-51 gene occurred in 235( 97.1%) samples. Most, 141 (58.2%) of the strains of Acinetobacter sp. They were resistant to three or more classes of antibiotics. A positive test for the detection of enzyme AmpC were 5.8% in Test Hodge was 61.1% and the ?-lactamase Metallo was 78.5% detection test. No samples proved producing ESBL. The genes most commonly found were the blaOXA-23 and blaOXA-143. The use of invasive devices (p = 0.001), the amount of antimicrobials used (p = 0.01), the ICU stay (p = 0.008) and hospitalization in a public hospital (p = 0.05) presented themselves as factors associated with the acquisition of Acinetobacter sp. strains MDR. Death was the most frequent clinical outcome (p <0.001) in patients with infection by these strains. The high number of multidrug-resistant Acinetobacter antimicrobial, isolated from patients and especially of inanimate surfaces in hospitals is disturbing. This scenario may compromise both the empirical treatment of seriously ill patients as the hospital infection control strategies.

As mudanças climáticas associadas às atividades humana são devidas principalmente às emissões de CO2 na atmosfera provenientes da queima de combustíveis de origem fóssil. Desta forma, faz-se necessária a substituição dessas fontes fósseis de geração de energia, por fontes renováveis. Dentre as alternativas de fontes renováveis, podemos destacar os biocombustíveis produzidos a partir de microalgas, as quais apresentam composição rica em óleos e proteínas. Um dos grandes desafios encontrados na conversão de biomassa em biocombustíveis é a caracterização detalhada das microalgas. A identificação de espécies através da espectrometria de massas com Ionização/Dessorção à Laser Assistida por Matriz acoplada a analisador por tempo de vôo (MALDI-TOF-MS) utilizada na análise de perfil de proteínas de micro-organismos, e posterior rápida identificação por comparação com os padrões armazenados em bancos de dados (fingerprint) tem se sobressaído. Existem poucos trabalhos na literatura abordando a identificação de espécies de microalgas utilizando a técnica de MALDI-TOF-MS e nenhum trabalho abordando a análise a partir do uso de células de microalgas liofilizadas. Desta forma, nesse trabalho foi estudada a influência de diversos parâmetros tais como placa, modo de análise, valor de PIE, valor de IS2, matriz e solvente de matriz e amostra nos espectros de massas do tipo MALDI-TOF-MS para análise do perfil proteico de células liofilizadas das espécies de microalgas Chlorella vulgaris, Chlorella sp., Desmodesmus sp., Monoraphidium sp. e Oocystis sp. Primeiramente, os cultivos foram realizados em um sistema de agitador orbital otimizado de tal maneira que todas as posições apresentassem as mesmas condições. Após os cultivos, as células foram secas para posterior análise de espectrometria de massas. Para determinação da metodologia que fornecesse os melhores espectros de massas, foram avaliados, aleatoriamente, 3 parâmetros: número de íons (P1), relação sinal/ruído do pico base (P2) e intensidade do pico base (P3). Foi observado que para a maioria das amostras de microalgas, os parâmetros que mais influenciaram na obtenção de espectros de massas do tipo MALDI-TOF bem resolvidos foram a placa, o modo de análise, valor de PIE, valor de IS2 e a matriz. As variações obtidas nos espectros de massas, quando utilizados diferentes solventes tanto para a matriz quanto para a amostra, bem como a adição de isopropanol com o objetivo de melhorar a distribuição da matriz sDHB na placa de amostragem, não foram tão significativas como as observadas para os outros parâmetros avaliados nesse estudo. Como conclusão, o uso da matriz sDHB, solvente TA50 para amostra e matriz, análise na placa polished sob as condições de análise PIE 100ns, IS2 23kV mostraram-se muito mais efetivos para a análise de proteínas a partir de amostras de microalgas liofilizadas. A análise dos lipídios apresentou uma distribuição predominante dos ácidos graxos C16:0, C18:2 e C18:0 para os cultivos de 12 dias e C16:0, C18:2 e C22:6 para os cultivos de 8 dias. Entretanto, as proporções de C22:6 e C18:2 aumentaram para os cultivos de 8 dias. Dessa forma, as espécies de microalgas Chlorella vulgaris., Chlorella sp., Monoraphidium sp. e Oocystis sp. cultivadas por 8 dias podem ser convertidas em biocombustível por apresentarem ácidos graxos entre 14 e 18 carbonos e em sua composição. / Climate change associated to human activities are mainly due to CO&#8322 emissions from combustion of fossil fuels in the atmosphere. Thus, it is necessary to replace these fossil sources of energy generation for renewable sources. Among the alternative of renewable sources, biofuels derived from microalgae is am potential alternative, since microalgae present in their composition fatty acids and proteins. Characterization of microalgae is one of the challenges in the conversion of their biomass into biofuels. The microorganism species identification by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF-MS) thought the analysis of protein profile and subsequent fast identification by comparison with the standard protein profile (fingerprint) in the database has been outstanding. There are few studies in the literature about the identification of microalgae species using MALDI-TOF-MS technique and there is no one using cells of lyophilized microalgae. Thus, in this work was studied the influence of many parameters such as target, analysis mode, PIE, IS2 value, matrix, matrix solvent and sample solvent in the MALDI-TOF mass spectra for analysis of protein profile of lyophilized microalgae cells for the species Chlorella vulgaris cells, Chlorella sp., Desmodesmus sp., Monoraphidium sp. and Oocystis sp. Cultivations were carried out using an optimized shaker system, where all positions presented the same conditions. After the cultivation, the cells were dried for subsequent mass spectrometric analysis. To achieve the best mass spectra profile, 3 parameters were arbitrarily evaluated: number of ions (P1), base peak signal/noise ratio (P2) and base peak intensity (P3). It was observed for most microalgae samples, MALDI-TOF mass spectra profile were most influenced by target, analysis mode, PIE value, IS2 value and the matrix. Variations in the mass spectra obtained when different solvents were used (for matrix and sample) as well as the addition of isopropanol in order to improve the distribution of sDHB matrix on the spot, were not significant as that observed for the other parameters. In conclusion, the use of sDHB matrix, TA50 solvent for sample and matrix, analysis in polished target plate under the following analysis conditions: a PIE 100ns, a IS2 23kV, provided to be more effective for the analysis of protein from lyophilized microalgae cells. Lipid analysis of 12 days cultivated microalgae showed a predominant distribution of the C16:0, C18:2 and C18:0. The 8 days cultivation presented a distribution of C16:0, C18:2 and C22:6, but with C22:6 e C18:2 in a higher proportion. Since biofuel are produced by using the C14-C18 fatty acid contained in their composition, 8 days cultivation showed to be most effective for this purpose.

Orientador: Marco-Aurelio De Paoli / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Quimica / Made available in DSpace on 2018-08-13T13:43:01Z (GMT). No. of bitstreams: 1 Romao_Wanderson_M.pdf: 3473303 bytes, checksum: 3ef4c09640bf5042dfcbfd4c110e4244 (MD5) Previous issue date: 2009 / Resumo: Atualmente, o Brasil apresenta um dos maiores índices mundiais de reciclagem mecânica do poli (tereftalato de etileno), PET. O sucesso desse termoplástico na indústria de reciclagem deve-se à sua ampla diversidade de aplicações. As embalagens recicladas grau alimentício podem ser misturadas com a resina virgem e reprocessadas. Três metodologias foram estudadas para detectar a presença do PET pós-consumo graugarrafa (PETpc-btg) em PET virgem grau-garrafa (PETv-btg): calorimetria exploratória diferencial (DSC), espectrometria de massas (MALDI-TOF MS) e Fluorescência de raios-X (XRF). Amostras de PETv-btg de três fabricantes foram analisadas: Braskem, Rhodia e Eastman. Amostras de PETpc-btg submetidas ao processo super-clean® também foram analisadas. Elas apresentam a mesma [h] do PETv-btg e foram fornecidas pela empresa Bahia PET. Amostras de PETv-btg Braskem e PETpc-btg foram misturadas e processadas em nosso laboratório em diversas proporções através de um misturador interno acoplado ao reômetro de torque. Os resultados de DSC mostram que a Tm, Tc, DCp e a cinética de cristalização são as principais propriedades térmicas que servem para diferenciar entre PETv-btg e PETpc-btg. Utilizando a técnica de MALDI-TOF MS aliado ao PCA (Análise Componentes Principais), foi possível distinguir as amostras em vários grupos. Esses grupos eram separados em função de alterações químicas como: variações na viscosidade intrínseca ([h] 0,80 e [h] = 0,65-60); submetidas e não submetidas a algum processo industrial; wt % de PETpc-btg em PETv-btg Braskem; e variação no processo de síntese do polímero (fabricante). A partir desses resultados foi possível construir um modelo de calibração, onde ele consegue distinguir entre uma amostra de PETv-btg e uma amostra de PETpc-btg. As medidas de XRF mostraram que alguns fabricantes utilizam mais de um catalisador para o processo de síntese do PETv-btg. A Braskem utiliza manganês e antimônio. Portanto, o modelo de previsão funciona para prever a wt % PETpc nas misturas que foram utilizadas na construção dele, como é o caso das resinas de PETv-btg Braskem e PETpc-btg. Observamos também, através das medidas de XRF, que o teor de Ferro presente no PET aumenta em função do processo de reciclagem. Esta variável poderá ser utilizada para a construção de um modelo quimiométrico abrangendo uma maior quantidade de variáveis / Abstract: Recently, Brazil recorded mechanical recycling of the poly (ethylene terepththalate), PET, the highest in the world, corresponding to about 53 wt %. This success in the recycling industry is due to the wide range of its applications, from textiles to packaging for the food industry. The recycled food-grade packaging could be mixed with virgin resin and reprocessing. Three methodologies were used to detect the presence of the bottle-grade post-consumption PET (PETpc-btg) in the bottle-grade virgin PET (PETv-btg): differential scanning calorimetry (DSC), x-ray fluorescence and matrix assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS). PETv-btg samples were supplied by the manufacturers (Braskem, Rhodia and Eastman). Samples from a superclean ® process (Bahia PET Reciclagem) were also analysed. All samples had the same intrinsic viscosity values, [h]. Braskem PETv-btg and PETpc-btg samples were blended and processed in different proportions in our laboratory using a HAAKE mixer system. DSC results show that cristallization kinetics, heat capacity (DCp), melting and crystallization temperature are the principal thermal properties that can be used to distinguish between PETv-btg and PETpc-btg. MALDI-TOF MS results together with PCA (principal component analysis) was used to classify the samples into several groups: intrinsic viscosity changes ([h] 0,80 e [h] = 0,65-60); processed and not submitted to some industrial process; wt % PETpc-btg in the PETv-btg Braskem; synthesis process change (manufacturer). From these results, it was possible to creat a calibration model, that differentiated between PETv-btg and PETpc-btg resins. However, we were not able to forecast the percentage of PETpc-btg in the PETv-btg. A model can be made from processed samples where its Mw could be corrected for solid state polymerization or the super-clean® process. XRF results show that some manufacturers use one or more catalysts for PETv-btg synthesis. The Braskem resin is made using manganese and antimony catalysts. Therefore, the prediction model is valid only when the origin of the studied mixture is known, such as PETv-btg/PETpc-btg processed blends. For other resins, the prediction model does not work. The Braskem resin had characteristics distintct from the others. We observed also that the Fe concentration in PET increase in as a function of the recycling process. Therefore, this variable could be used, in the future work, to create chemometric models incluing a higher number of variables / Mestrado / Físico-Química / Mestre em Química

Submitted by Elizete da Silva Dias (elizetedias@ufam.edu.br) on 2017-06-22T15:28:23Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) TESE - ANDREIA FERREIRA DE SOUZA.pdf: 2861845 bytes, checksum: fa54456b6bc990f480efa541a3b7cdfe (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-06-23T14:30:14Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) TESE - ANDREIA FERREIRA DE SOUZA.pdf: 2861845 bytes, checksum: fa54456b6bc990f480efa541a3b7cdfe (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-06-23T14:32:11Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) TESE - ANDREIA FERREIRA DE SOUZA.pdf: 2861845 bytes, checksum: fa54456b6bc990f480efa541a3b7cdfe (MD5) / Made available in DSpace on 2017-06-23T14:32:11Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) TESE - ANDREIA FERREIRA DE SOUZA.pdf: 2861845 bytes, checksum: fa54456b6bc990f480efa541a3b7cdfe (MD5) Previous issue date: 2016-11-18 / Item withdrawn by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-07-14T13:45:46Z Item was in collections: Doutorado (ID: 91) No. of bitstreams: 4 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Reprodução Não Autorizada.pdf: 47716 bytes, checksum: 0353d988c60b584cfc9978721c498a11 (MD5) Reprodução Não Autorizada.pdf.txt: 65 bytes, checksum: 84fdbdbfeca4f7e4764b2f14d97a84ad (MD5) Reprodução Não Autorizada.pdf.jpg: 2921 bytes, checksum: b6f7c946b94a5bc7f226fe694f68a640 (MD5) / The C. albicans is the most frequently isolated fungus in humans healthy, being considered the main opportunistic pathogenic species, However, in recent decades, it has been observed a significant increase of other non-albicans species, involved both in colonization as in human infectious processes. The clinical manifestation but prevalent, This fungus, the woman is Vulvovaginal candidiasis (VVC). This study had the objective of making the molecular characterisation and polyphasic taxonomy of Candida species isolated from vaginal exsudate and describe their susceptibility profiles antifungals and extracts of Vatairea guianensis. The identification was performed by methods Classics (Microcultive, CHROMagar, Auxonograme), MALDI-TOF, multiplex PCR and molecular. In the analysis of genetic variability was made to AFLP.The colonization of Candida sp. in the vagina was statistically significant in patients aged 35 years or more, which made use of underwear fair and/or synthetic, in use of antibiotic therapy and who had CVV recurrent episodes. Of the 202 samples 37.1% (N = 75) was possible isolating yeasts Candida sp. C. albicans was the most isolated species (64%), followed by C. parapsilosis (20%), C. glabrata (5.3%), C. tropicallis (4%), C. haemolinus (4%) and C. krusei (1.3%) and C. lusitaniae (1.3%). Combinations of methods are required phenotypic and also carrying out additional evidence to the identification of Candida spp. molecular methods available confirm that the use phenotypic methods for clinical diagnosis must be cautious, have low accuracy in determination of Candida species not albicans and the identification of new species. The MALDI-TOF proved as an excellent tool for species identification of Candida sp, as it associates simple technique, procedures partially automated and mainly the speed of the result Microbiology, the limitation of this method is in the absence of a complete database. C. haemulonii, an emerging and rare species, was first identified in isolates of vaginal exudate asymptomatic women in Manaus. The vast majority of isolates of Candida sp., vaginal exudate, presented resistance to antifungals fluconazole, itraconazole, ketoconazole and Nystatin. The extracts of the seeds of V. guianensis presented only fungistatica activity in 04 isolates of Candida sp. vaginal exsudate. Thus, this research contributed to the epidemiological knowledge of species of Candida in vaginal flora of assintomaticas patients, and may guide the diagnosis in pathological conditions and being used in monitoring of emerging infections and drug resistant, because the variation of species according to geographical location should be taken into account between the epidemiological factors of CVV. / A C. albicans é o fungo mais frequentemente isolado em humanos sadios, sendo considerada a principal espécie patogênica oportunista, entretanto, nas últimas décadas, tem sido observado um significativo aumento de outras espécies não-albicans, implicadas tanto na colonização como em processos infecciosos humanos. A manifestação clínica mais prevalente, deste fungo, na mulher é a Candidíase Vulvovaginal (CVV). OBJETIVOː realizar a caracterização molecular e a taxonomia polifásica de espécies de Candida oriundas de isolados de exsudato vaginal e descrever seus perfis de susceptibilidade a antifúngicos e aos extratos de Vatairea guianensis. METODOLOGIAː A identificação foi realizada por métodos clássicos (Microcultivo, CHROMagar, Auxonograma), MALDI-TOF, multiplex PCR e a molecular. A variabilidade fenotipica foi feita por AFLP, e os testes de susceptibilidade em placa de Elisa por microdiluição com os antifungicos e os extratos de Vatairea guianensis. RESULTADOː A colonização de Candida sp. na vagina foi estatisticamente significativa em pacientes com idade igual ou maior que 35 anos, que faziam uso de roupas íntimas justas e/ou sintéticas, em uso de antibioticoterapia e que tinham episodios CVV recorrente. Das 202 amostras 37,1 % (N=75) foi possível isolar leveduras Candida sp. Na analise da variabilidade genética foi feito a AFLP. Candida albicans foi a espécie mais isolada (64 %), seguida por C. parapsilosis (20 %), C. glabrata (5,3 %), C. tropicallis (4 %), C. haemolinus (4%) e C. krusei (1,3 %) e C. lusitaniae (1,3 %). São necessárias combinações entre métodos fenotípicos e também a realização de provas complementares para a identificação de Candida spp. Os métodos moleculares disponíveis confirmam que o uso de métodos fenotípicos para o diagnóstico clínico deve ser cauteloso, pois possuem baixa acurácia na determinação de espécies Candida não albicans e na identificação de novas espécies. CONCLUSÃOː O MALDI-TOF se mostrou como uma excelente ferramenta para a identificação de espécies de Candida sp, pois associa simplicidade da técnica, procedimentos parcialmente automatizados e principalmente a rapidez do resultado microbiológico, a limitação deste método esta na ausência de um completo banco de dados. C. haemulonii, uma espécie emergente e rara, foi pela primeira vez identificada em isolados do exsudato vaginal de mulheres assintomáticas em Manaus. A grande maioria dos isolados de Candida sp., do exsudato vaginal, apresentou resistência aos antifúngicos fluconazol, itraconazol, cetoconazol e a nistatina. Os extratos das sementes de Vaitarea guianensis apresentou apenas atividade fungistatica em 04 isolados de Candida sp. do exsudato vaginal. Assim, esta pesquisa contribuiu para o conhecimento epidemiológico das espécies de Candida na flora vaginal de pacientes assintomaticas, podendo orientar o diagnóstico em condições patológicas e sendo utilizado na monitorização de infecções emergentes e resistentes às drogas, pois a variação das espécies de acordo com a localização geográfica deve ser levada em conta entre os fatores epidemiológicos de CVV.

Submitted by Kamila Costa (kamilavasconceloscosta@gmail.com) on 2015-07-14T19:57:04Z No. of bitstreams: 1 Dissertação - Silmara M Miranda.pdf: 1535464 bytes, checksum: 2e2afc8694fa41a1cb6af9490965b12b (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-16T19:32:15Z (GMT) No. of bitstreams: 1 Dissertação - Silmara M Miranda.pdf: 1535464 bytes, checksum: 2e2afc8694fa41a1cb6af9490965b12b (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-16T19:36:56Z (GMT) No. of bitstreams: 1 Dissertação - Silmara M Miranda.pdf: 1535464 bytes, checksum: 2e2afc8694fa41a1cb6af9490965b12b (MD5) / Made available in DSpace on 2015-07-16T19:36:56Z (GMT). No. of bitstreams: 1 Dissertação - Silmara M Miranda.pdf: 1535464 bytes, checksum: 2e2afc8694fa41a1cb6af9490965b12b (MD5) Previous issue date: 2014-06-27 / FAPEAM - Fundação de Amparo à Pesquisa do Estado do Amazonas / Filamentous fungi are organisms with the ability to produce various secondary metabolites, among them, mycotoxins, characterized by the toxic properties that have the human and animal organism. Are produced mostly by species of the genera Aspergillus and Penicillium, by engaging with relevant food contamination and mycotoxin production. The study aimed to identify the mycoflora and mycotoxins in samples of cassava produced in TNL in the Amazon region, Brazil flour. 30 samples of cassava flour in the local market and evaluated for moisture and water activity (aw), fungi (isolation, purification and identification by traditional taxonomy) and mycotoxins were collected. Confirmation of toxigenic fungal species was made by molecular biology, scanning electron microscopy was performed in three toxigenic species and mycotoxins were identified by MALDI-TOF. The results of moisture and (aw) ranging from 7.08 to 13.55% and 0.37 to 0.69 respectively. Among the samples analyzed, 30% had fungi and of these 18.5% were toxigenic. The species most frequently identified were Aspergillus flavus, Penicillium waksmanii and Penicillium citrinum. Among the toxigenic fungi Penicillium citrinum, Aspergillus flavus and Aspergillus amoenus were found. MALDI-TOF showed the presence of aflatoxin B1, patulin and citrinana more frequently. We conclude that greater attention to cassava flour, one of the most consumed foods in the north of the country is needed as toxigenic fungi and mycotoxins were found in the samples analyzed. / Fungos filamentosos são organismos com a capacidade de produzir diversos metabólitos secundários, dentre eles, as micotoxinas, caracterizadas pelas propriedades tóxicas que apresentam ao organismo humano e animal. São produzidas em sua maioria por espécies dos gêneros Aspergillus e Penicillium, relevantes pelo envolvimento com contaminação de alimentos e produção de micotoxinas. O estudo objetivou identificar a micobiota e ocorrência de micotoxinas em amostras de farinha de mandioca produzidas em Coari na região Amazônica, Brasil. Foram coletadas 30 amostras de farinha de mandioca no mercado local e avaliadas quanto à umidade e atividade de água (aw), fungos (isolamento, purificação e identificação pela taxonomia tradicional) e micotoxinas. A confirmação das espécies fúngicas toxigênicas foi feita pela biologia molecular, foi realizada microscopia eletrônica de varredura em três espécies toxigênicas e micotoxinas foram identificadas por MALDI-TOF. Os resultados de umidade e (aw) variaram de 7,08 a 13,55% e 0,37 a 0,69 respectivamente. Entre as amostras analisadas, 30% apresentaram fungos e destes 18,5% foram toxigênicos. As espécies identificadas com maior frequência foram Aspergillus flavus, Penicillium waksmanii e Penicillium citrinum. Entre os fungos toxigênicos foram encontrados Penicillium citrinum, Aspergillus flavus e Aspergillus amoenus. MALDI-TOF demonstrou presença de aflatoxina B1, citrinana e patulina com maior frequência. Conclui-se que é necessário maior orientação para os produtores da farinha de mandioca, um dos alimentos mais consumido na região Norte do país, pois fungos toxigênicos e micotoxinas foram encontrados nas amostras analisadas.

Submitted by Rosangela Silva (rosangela.silva3@unioeste.br) on 2018-05-24T20:32:41Z No. of bitstreams: 2 Charles Seuchuco.pdf: 1461597 bytes, checksum: d1fe07c8c4b16541956c362cc1acb19c (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-05-24T20:32:41Z (GMT). No. of bitstreams: 2 Charles Seuchuco.pdf: 1461597 bytes, checksum: d1fe07c8c4b16541956c362cc1acb19c (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-03-05 / Diatraea saccharalis, known as a sugarcane borer, is a moth of the Lepidoptera order, being one of the main pests on sugarcane, responsible for damage to the sugar and alcohol industries. The fat body is the organ responsible for several functions during the larval phase of insects, mainly in charge of synthesizing molecules with metabolic actions. The success in insects’ adaptation is due to its efficient defense system, in which the fat body is able to synthesize antimicrobial peptides (AMP’s) and secrete them in the hemolymph, where they act as a defense mechanism against pathogens. AMP’s has a cationic character and present the capacity to interact with negative charge molecules of the bacteria, leading to the disintegration of their membranes. Considering the concerns regarding bacterial resistance to antibiotics and antimicrobial properties of AMP’s, such as fast-acting, varied mechanisms of action and wide range against Gram-positive and Gram-negative bacteria; it has become appropriate to carry out studies to use AMP’s as alternative or complementary antibiotic. In D. saccharalis, AMP’s with antimicrobial action have been reported in recently published papers using proteomic methodology. Thereby, this study aimed to perform the differential analysis of the proteins and peptides of the fat body in the 5th instar of larvae challenged with Escherichia coli, Bacillus subtilis and Beauveria bassiana; using the two-dimensional electrophoresis technique (2-DE), with 12.5% SDS-PAGE tricine gels, MALDI-TOF type mass spectrometry and the research in protein databases (Mascot and TagIdent). After analysis, six defense proteins of the innate immune system were found using the TagIdent identification tool: apolipophorin-3, attacin, attacin-F, Peptidoglycan recognition protein (PGRPs), putative defense protein 1 and putative defense protein 3. The AMP attacin presented a positive regulation in relation to the control gel indicating that the B. bassiana septic challenge stimulated the immune system of D. saccharalis. This study provides the first reports of proteins of the immune system produced by the adipose tissue of D. saccharalis. / A Diatraea saccharalis, conhecida como broca-da-cana, é uma mariposa da ordem Lepidoptera, tratando-se de uma das principais pragas da cana-de-açúcar, responsável por prejuízos à indústria de açúcar e de álcool. O corpo gorduroso é o órgão responsável por várias funções durante a fase larval dos insetos, principalmente encarregado de sintetizar moléculas com ações metabólicas. O sucesso na adaptação dos insetos se deve ao seu eficiente sistema de defesa, no qual o tecido gorduroso é capaz de sintetizar peptídeos antimicrobianos (PAM’s) e secretá-los na hemolinfa, onde atuam como mecanismo de defesa contra patógenos. Os PAM’s têm caráter catiônico e apresentam capacidade de interagir com as moléculas de carga negativas das bactérias, levando a desintegração de suas membranas. Considerando as preocupações em relação à resistência bacteriana a antibióticos e às propriedades antimicrobianas dos PAM’s como ação rápida, mecanismos de ação variados e amplo espectro contra bactérias Gram-positivas e Gram-negativas, têm se tornado atrativo para a realização de estudos para utilização de PAM’s como antibiótico alternativo ou complementar. Em D. saccharalis já foram relatados PAM’s com ação antimicrobiana em artigos recentemente publicados empregando metodologia proteômica. Desta forma, este trabalho teve como objetivo realizar a análise diferencial das proteínas e peptídeos do corpo gorduroso no 5° instar de larvas controles e desafiadas com Escherichia coli, Bacillus subtilis e Beauveria bassiana, utilizando eletroforese bidimensional (2-DE) com géis tricina SDS-PAGE 12,5%, espectrometria de massas do tipo MALDI-ToF e a pesquisa em bancos de dados de proteínas (Mascot e TagIdent). Após análise, foram encontradas seis proteínas de defesa do sistema imune inato utilizando a ferramenta de identificação TagIdent, sendo elas Apolipophorin-3, Atacina, Atacina-F, Proteína de reconhecimento de peptidoglicano (PGRPs), Provável proteína de defesa 1 e Provável proteína de defesa 3. O PAM Atacina apresentou regulação positiva em relação ao gel controle, indicando que o desafio séptico por B. bassiana estimulou o sistema imune da D. saccharalis. Este trabalho fornece os primeiros relatos de proteínas do sistema imune produzidas pelo corpo gorduroso de D. saccharalis.

La composition chimique de la résine sandaraque et de son composant principal l’acide communique a été étudiée par chromatographie en phase gazeuse – spectrométrie de masse (GC-MS), MALDI-TOF (désorption-ionisation laser assistée par matrice - temps de vol), ESI (ionisation par électronébuliseur) - Orbitrap, FTIR/ATR (spectroscopie infrarouge à transformée de Fourier/réflectance totale atténuée), spectroscopie de RMN (résonance magnétique nucléaire) à l'état solide et liquide. Six composés avec des squelettes labdane et pimarane ont été identifiés dans la résine commerciale. Les spectres de masse obtenus ont été interprétés et le comportement en spectrométrie de masse de ces diterpénoïdes dans les conditions de l’impact électronique a été décrit. L'analyse quantitative par la méthode de l'étalon interne a révélé que les diterpénoïdes identifiés ne représentaient que 10 à 30% de l'échantillon analysé. La complexité de la fraction réticulée de la résine commerciale sandaraque est bien reflétée par les spectres de masse MALDI-TOF et ESI-Orbitrap. En conséquence, les spectres de masse de MALDI-TOF comprenaient trois clusters de pics dans la gamme m/z de 300-900, et ceux d’ESI-Orbitrap contenaient cinq clusters de pics dans la gamme m/z de 300-1100. Les pics dans les clusters correspondent aux dérivés oxygénés des diterpénoïdes. Les résultats obtenus à partir des expériences RMN par IRCP (Inversion Recovery Cross-Polarization) ont révélé le caractère rigide des échantillons de la résine sandaraque analysés et justifiaient l'hypothèse que le reste de l'échantillon, qui ne pouvait être quantifié par la méthode de l'étalon interne, aurait un caractère polymère. / The chemical composition of sandarac resin and its principal component communic acid was investigated by gas chromatography-mass spectrometry (GC-MS), MALDI-TOF (Matrix Assisted Laser Desorption Ionization - Time of Flight), ESI (Electrospray ionization)-Orbitrap, FTIR/ATR (Fourier transform infrared spectroscopy/Attenuated total reflectance), liquid- and solid state NMR (Nuclear magnetic resonance) spectroscopy. Six compounds with labdane and pimarane skeletons were identified in the commercial resin. The obtained mass spectra were interpreted and the mass spectrometric behaviour of these diterpenoids under EI conditions was described. Quantitative analysis by the method of internal standard revealed that identified diterpenoids represent only 10–30% of the analysed sample. The complexity of the reticulated fraction of the commercial sandarac resin was well reflected by the MALDI-TOF and ESI-Orbitrap mass spectra. As a result, MALDI-TOF mass spectra comprised three clusters of peaks in the m/z range of 300–900, and for the ESI-Orbitrap mass spectra contained five clusters of peaks in the m/z range of 300–1100. The peaks in the clusters corresponded to the oxygenated derivatives of the diterpenoids. The results obtained from the IRCP (Inversion Recovery Cross-Polarization) experiments revealed the rigid character of the sandarac resin samples analyzed and justified the hypothesis that the rest of the sample, which could not be quantified by the method of internal standard, would have a polymeric nature.

>Magister Scientiae - MSc / Salvia hispanica L. is an annual pseudocereal food crop, locally known as chia that has the ability to grow in water stress environments. The importance of chia dates back to the pre-columbian era where it was consumed as staple food by the indigenous South Americans due to its high nutritional and medicinal benefits. A single chia plant produces two seed variants: white seed genotype (denoted as WSG) and black seed genotype (denoted as BSG). Chia seeds have been proven to have a huge potential as a healthy food source and contained various medicinal properties. However, these plants are still prone to environmental stress conditions such as salinity that is one of the major abiotic stresses that influence crop production and yield worldwide. Despite the nutritional impact of the chia seeds, limited information regarding their molecular responses to abiotic stress conditions are known. This study was divided into two distinct parts. Firstly, the study comparatively analysed the leaf proteomes of two chia genotypes using gelbased proteomic analysis coupled with mass spectrometry. Total soluble proteins were extracted from chia leaves and subjected to 2-D PAGE analysis. Proteins were visualized by CBB and identified by MALDI-TOF MS/MS. A total of 284 and 209 spots were detected in WSG and BSG, respectively. Using mass spectrometry, 36 differentially expressed protein spots were successfully identified based on their protein abundance using homology database searches. Interestingly, two defensive-related proteins (osmotin-like protein and the chalcone isomerase) were only present in WSG and absent in BSG. In light of previous information regarding the nutritional profiles (no significant difference) of these two genotypes, this study has shown that there are distinct molecular differences between these genotypes. Therefore, WSG will be used in further downstream analysis. The second part of this study focused on the influence of salt stress (imposed by 100 mM NaCl) on the leaf proteome of WSG. Using gel-based proteomic analysis, 61 differentially expressed proteins were identified and classified into nine functional categories. Most of the proteins identified in this study were upregulated by salt stress. Interesting to note, 12 proteins identified in this study were only present in response to salt stress but were absent in the control. These proteins include ATP-dependent zinc metalloprotease FTSH 2 (spot 48), HSP70 proteins (spots 46 and 47), superoxide dismutases (spots 10, 41 and 42) and an ascorbate peroxidase (spot 56). All these proteins are important antioxidants that play a significant role in scavenging reactive oxygen species (ROS). Previous studies have shown that these antioxidants play vital roles in stress tolerance. These proteins could serve as potential biomarkers that could be used to enhance salt stress tolerance in pseudocereals and cereal food crops. / National Research Foundation (NRF) and Agricultural Research Council