Submaximal exercise performance is, in part, limited by the accumulation of metabolic byproducts and energy system capacities. Curcumin and the combination of curcumin and fenugreek soluble fiber (CurQfen®) have been shown to increase endogenous antioxidants and metabolic byproduct clearance as well as reduce inflammation and lipid peroxidation, and therefore, may enhance submaximal aerobic thresholds. In addition, there is evidence that the galactomannan component of fenugreek, used to enhance bioavailability of curcumin, may also have potential physiological effects related to the up regulation of free fatty acid oxidation Therefore, the purpose of this study was to examine the effects of curcumin and fenugreek soluble fiber supplementation on the ventilatory threshold (VT), respiratory compensation point (RCP), maximal oxygen consumption (O2 peak), and time to exhaustion (Tlim)derived from a graded exercise test (GXT). Forty-five untrained, college-aged, male (n = 24) and female (n = 21) subjects (mean age ± SD: 21.2 ± 2.5 yr) were randomly assigned to one of three supplementation groups; placebo (PLA, n=13), 500 mg·day-1 CurQfen® (CUR, n=14), or 300 mg·day-1 fenugreek soluble fiber (FEN, n=18). All of the subjects completed a maximal GXT on a cycle ergometer to determine the VT, RCP, O2 peak, and Tlim before (PRE) and after (POST) 28 days of daily supplementation. The VT and RCP were determined from the V-slope method for the ventilation (E)vs. O2 and E vs. CO2, respectively. Separate, one-way ANCOVAs were used to examine the between group differences for adjusted POST VT, RCP, O2 peak, and Tlim values, with the respective PRE test value as the covariate. The adjusted POST VT-O2 for the CUR (mean ± SD= 1.593 ± 0.157 L·min-1) and FEN (1.597 ± 0.157L·min-1) groups were greater than (p= 0.04 and p= 0.03, respectively) the PLA (1.465 ± 0.155L·min-1) group, but the FEN and CUR groups were not different (p = 0.94). The one-way ANCOVAs for RCP (F = 3.177, p = 0.052), O2 peak (F = 0.613, p = 0.547), and Tlim (F = 0.654, p = 0.525) indicated there were no significant differences among groups. These findings suggested that CurQfen® and/or fenugreek soluble fiber may improve submaximal, but not maximal, aerobic performance indices in untrained subjects.

Submitted by admin tede (tede@pucgoias.edu.br) on 2017-04-27T14:24:41Z No. of bitstreams: 1 DAIANE DE OLIVEIRA CUNHA.pdf: 2074632 bytes, checksum: 76e82c4169a9d294dcddcc8e9ad849cb (MD5) / Made available in DSpace on 2017-04-27T14:24:41Z (GMT). No. of bitstreams: 1 DAIANE DE OLIVEIRA CUNHA.pdf: 2074632 bytes, checksum: 76e82c4169a9d294dcddcc8e9ad849cb (MD5) Previous issue date: 2017-03-13 / The present dissertation was constructed in the form of scientific articles, being constituted by two articles. The first one, titled "Epidemiological evaluation of patients submitted to the serum galactomannan test with suspected Aspergillosis Invasive", aimed to evaluate the incidence of Invasive Aspergillosis in patients with hematological diseases followed at a reference hospital in Goiânia, Goiás, Brazil. Determine the main factors that contribute to the development of the disease. For this, 1367 samples of 264 patients with malignant hematological diseases treated in the Hematology Sector of Hospital Araújo Jorge, who underwent galactomannan detection, were evaluated in the period from 2013 to 2015, and were excluded patients who had no clinical data in Medical records and loss of follow-up at the hospital. Among the data obtained through the present study it was observed that the mean age was 43.7 years, where 55.5% were male. Of the total number of patients evaluated, 133 performed BMT, 38.9% of which were autologous and 17.9% of the allogeneic type. According to the classification for Invasive Aspergillosis according to the European Organization for Research and Treatment of Cancer, the disease was interpreted as proven in 7.3%, defined by culture positive for the fungus, 6.4% as probable by the detection of galactomannan in the Blood and presence of pulmonary infiltrates and 5.1% as possible by radiological alterations suggestive of Invasive Aspergillosis and galactomannan negative. The estimated / probable / probable Aspergillosis Invasive mortality rate was 61.3% and showed that the disease was significantly associated with the risk of death (p <0.0001). When considering the high mortality rate caused by the development of Aspergillosis Invasive and the fact that an early therapy promotes a significant improvement in the prognosis of patients, we conclude that the detection of galactomannan, performed as a follow-up of patients at high risk of developing the disease, can Be considered an effective method to aid in the identification of Invasive Aspergillosis. The second article entitled "Association between polymorphisms of genes encoding Dectin-1 and Toll-like cellular receptors and susceptibility to Invasive Aspergillosis", aimed to describe the polymorphisms in genes encoding Dectin-1 and Tolllike receptors, Seeking possible associations with the individual susceptibility to Invasive Aspergillosis. This study used as a methodological basis a systematic review of the literature and as a database to PubMed and PMC of the NCBI. The keywords used were: Invasive aspergillosis, polymorphism, Dectin-1 and Toll-like. From this search, 415 studies were found and according to the inclusion and exclusion criteria 8 studies were selected. With the accomplishment of this, it was verified that several are the studies that describe the single base polymorphisms with a greater susceptibility to the Invasive Aspergillosis. The major ones are in genes encoding Toll-like receptors and those that encode cytokines and chemokines (Dectin-1). Thus, it can be observed that, according to studies already carried out, there is a significant association of genetic polymorphisms with Aspergillosis Invasive, but more extensive studies must be performed to obtain more reliable results. In general, we conclude with this dissertation that the development of Invasive Aspergillosis is associated with several factors, one of them being genetic, and that, due to the high mortality rates caused by Invasive Aspergillosis, the detection of galactomannan is of fundamental importance for the Treatment of patients who are more likely to develop the disease, mainly aiding in the prognosis and early diagnosis of it. / A presente dissertação foi construída na modalidade de artigos científicos, sendo constituída por dois artigos. O primeiro, intitulado “Avaliação epidemiológica de pacientes submetidos ao teste de galactomanana sérica com suspeita de Aspergilose Invasiva”, teve como objetivo avaliar a incidência de Aspergilose Invasiva em pacientes com doenças hematológicas acompanhadas em um hospital de referência de Goiânia, Goiás, Brasil, e determinar os principais fatores que contribuem para o desenvolvimento da doença. Para isso, foram avaliadas 1367 amostras de 264 pacientes com doenças hematológicas malignas tratadas no Setor de Hematologia do Hospital Araújo Jorge, os quais realizaram exame de detecção de galactomanana, no período de 2013 a 2015, e foram excluídos pacientes que não possuíam dados clínicos em prontuários e com perda de seguimento no hospital. Dentre os dados alcançados por meio do presente estudo observou-se que a média de idade foi de 43,7 anos, onde 55,5% eram pertencentes ao sexo masculino. Do total de pacientes avaliados, 133 realizaram TMO, sendo que 38,9% eram do tipo autólogo e 17,9% do tipo alogênico. De acordo com a classificação para Aspergilose Invasiva conforme a European Organization for Research and Treatment of Cancer, a doença foi interpretada como comprovada em 7,3%, definida por cultura positiva para o fungo, 6,4% como provável pela detecção de galactomanana no sangue e presença de infiltrados pulmonares e 5,1% como possível por alterações radiológicas sugestivas de Aspergilose Invasiva e galactomanana negativo. A taxa de mortalidade para Aspergilose Invasiva comprovada/provável/possível foi de 61,3% e mostrou que a doença estava significativamente associada com o risco de morte (p<0,0001). Ao considerarmos a alta taxa de mortalidade causada pelo desenvolvimento da Aspergilose Invasiva e que a realização de uma terapia precoce promove significativa melhora do prognóstico dos pacientes, concluímos que a detecção de galactomanana, realizada como acompanhamento dos pacientes com alto risco de desenvolver a doença, pode ser considerada um método eficaz para auxiliar na identificação de Aspergilose Invasiva. O segundo artigo, intitulado “Associação entre polimorfismos dos genes que codificam os receptores celulares Dectina-1 e Toll-like e susceptibilidade à Aspergilose Invasiva”, teve como objetivo descrever os polimorfismos nos genes que codificam os receptores Dectina-1 e Toll-like, buscando possíveis associações com a susceptibilidade individual à Aspergilose Invasiva. Este estudo utilizou como base metodológica uma revisão sistemática da literatura e como base de dados a PubMed e PMC do NCBI. As palavras-chaves utilizadas foram: Invasive aspergillosis, polymorphism, Dectin-1 e Toll-like. A partir desta busca, 415 estudos foram encontrados e de acordo com os critérios de inclusão e exclusão 8 estudos foram selecionados. Com a realização deste, constatou-se que vários são os estudos que descrevem os polimorfismos de base única com uma maior susceptibilidade à Aspergilose Invasiva. Os principais são em genes que codificam os receptores Toll-like e os que codificam citocinas e quimiocinas (Dectina-1). Assim, pode-se perceber que de acordo com estudos já realizados há uma associação significativa de polimorfismos genéticos com a Aspergilose Invasiva, porém estudos mais amplos devem ser realizados para se obter resultados mais fidedignos. De modo geral, concluímos com essa dissertação que o desenvolvimento da Aspergilose Invasiva está associado com diversos fatores, sendo um deles o genético, e que, devido aos altos índices de mortalidade causados pela Aspergilose Invasiva, a detecção de galactomanana é de fundamental importância para o tratamento dos pacientes que apresentam uma maior probabilidade de desenvolver a doença, auxiliando principalmente no prognóstico e diagnóstico precoce da mesma.

Estudos recentes têm sugerido que doença pulmonar obstrutiva crônica (DPOC) possa ser um fator de risco para aspergilose invasiva (AI), particularmente no contexto de ventilação mecânica e uso de esteróides. Neste trabalho, realizamos estudo de coorte prospectivo multicêntrico (2009-2010) em três unidades de terapia intensiva no Sul do Brasil. Foram incluídos no estudo pacientes com DPOC que apresentassem novo infiltrado pulmonar enquanto em ventilação mecânica e sob uso de corticosteróides. Para estes pacientes, foram realizados os seguintes testes, em amostras respiratórias (maioria aspirado traqueal): exame micológico direto, cultura quantitativa para fungos, pesquisa de antígeno galactomanana (GM) (Platelia Aspergillus) e PCR em tempo real para Aspergillus. O DNA das amostras respiratórias foi extraído utilizando-se o kit de extração MycXtra (Myconostica, UK), sendo a amplificação feita com dois kits comerciais de q-PCR: Aspergillus spp q-PCR Alert kit (Nanogen, Itália) e MycAssayTM Aspergillus kit (Myconostica, UK). Foi também obtido soro destes pacientes, onde foi testada GM, precipitinas para Aspergillus e IgE total. O estudo foi aprovado no comitê de ética dos dois hospitais. Foram incluídos no estudo 47 pacientes (40,4% do sexo masculino), sendo a idade média de 68,6 anos (±9,9). A maioria (72,8%) dos pacientes possuía DPOC grave (GOLD III/IV). A dosagem de esteróides (equivalentes de prednisona) variou de 100-4125 mg (mediana: 900 mg). Exame micológico (direto e cultivo) foi positivo para Aspergillus seção Fumigatti em apenas dois pacientes (4,2%). Outros fungos identificados foram Scedosporium apiospermum (n=1) e Histoplasma capsulatum (n=1). Precipitinas para Aspergillus foram positivas em três pacientes, com títulos baixos (<1:2). Os níveis de IgE variaram de 2 a >3000 UI/ml (mediana de 74 UI/ml). Em sua grande maioria, os índices de GM no soro foram <0,5, enquanto que nas amostras respiratórias, os índices de GM foram >0,5, >1,0 e >1,5 em 74,5%, 40,5% e 21,3%, respectivamente. PCR da Myconostica foi positivo em 10 pacientes, enquanto PCR Nanogen detectou apenas um paciente. A mortalidade geral foi de 53,2%. Este estudo prospectivo multicêntrico mostrou uma baixa incidência (4,2%) de AI em pacientes com DPOC. A determinação de GM mostrou altos índices nas amostras analisadas (50% com índices ópticos >1,3), possivelmente necessitando um maior ponto de corte para excluir resultados falso-positivos. A combinação de PCR e GM para o diagnóstico de AI em amostras respiratórias merece investigação adicional, devido à baixa sensibilidade dos métodos de cultivo observados nos estudos clínicos realizados. / Recent data have suggested that chronic obstructive pulmonary disease (COPD) may be an important risk factor for invasive aspergillosis (IA), particularly in the context of mechanical ventilation (MV) and therapy with corticosteroids. Here we present the results of a prospective multicentric study (2009-2010) conducted in three intensive care units (ICUs) in Southern Brazil. COPD patients on steroids showing a new lung infiltrate while on mechanical ventilation were included and the following tests were performed in respiratory samples (mostly tracheal aspirates): microscopy, quantitative fungal culture, galactomannan (GM) (Platelia Aspergillus EIA) and real-time PCR to detect Aspergillus DNA. DNA was extracted using MycXtra kit (Myconostica, UK) and amplification was performed using two q-PCR commercial kits: Aspergillus spp q-PCR Alert kit (Nanogen, Italy) and MycAssayTM Aspergillus kit (Myconostica, UK). Serum was also obtained and tested for Aspergillus precipitins, GM and total IgE levels. Ethical approval was obtained in each of the participant hospitals. A total of 47 patients were enrolled in the study (male 59.6%). Mean age was 68.6 years-old (± 9.9). Most patients had severe COPD (GOLD stages III/IV in 72.8%). Steroid dosage (prednisone equivalent) ranged from 100-4125 mg (median 900 mg). Microscopy and culture were positive for Aspergillus section Fumigatti in only 2 patients (4.2%). Other fungi included H. capsulatum (n=1) and S. apiospermum (n=1). Aspergillus precipitins were positive for three patients, at low titers (<1:2). IgE levels ranged from 2 to >3,000 IU/ml (median 74 IU/ml). All serum GM indexes were <0.5 and respiratory samples, GM indexes of >0.5, >1.0 and >1.5 were observed in 74.5%, 40.5%, and 21.3%, respectively. Myconostica PCR was positive in 10 patients, while Nanogen PCR detected only one patient. Overall mortality was 53.2%. This prospective multicenter study showed a low incidence (4.2%) of IA in critically ill patients with COPD. High optical indices were observed when GM was tested in respiratory samples (50% of the results showed indices of >1.3). Therefore, the test did not discriminate IA and a a higher cutoff would be needed to exclude false-positive results. The combination of PCR and GM for the diagnosis of IA in respiratory samples deserves further investigation due to the low diagnostic sensitivity of the classical mycology methods.

Laspergillose naso-sinusale (ANS) est une des causes les plus fréquentes de jetage nasal chez le chien. Cette affection peut être fortement suspectée sur la base du signalement, de lanamnèse et de lexamen clinique. Cependant, la confirmation du diagnostic requiert la combinaison de divers examens complémentaires dont lendoscopie et limagerie médicale en coupe, deux techniques principalement réservés aux centres de médecine vétérinaire spécialisés. Il serait pourtant utile pour le vétérinaire praticien de pouvoir poser lui-même le diagnostic grâce à une méthode alternative fiable et peu/non-invasive. Cest pourquoi, nous avons, dans la première partie de ce travail, investigué plus en profondeur lutilité de deux types de prélèvements faciles à prélever pour le vétérinaire (du sang et des sécrétions nasales) dans le diagnostic de lANS. Lors dune première étude, nous avons investigué lintérêt de la détection danticorps spécifiques dAspergillus dans le sérum par deux tests sérologiques (un test dimmunodiffusion sur gel dagarose (AGDD) et un test ELISA IgG indirect) en utilisant une nouvelle solution commercialisée dantigènes purifiés dAspergillus. Nous avons également investigué la valeur diagnostique de la détection de galactomannan (antigènes dAspergillus) (GM) dans le sérum au moyen dun test ELISA (PlateliaTM Aspergillus). Alors que les tests sérologiques (AGDD et ELISA IgG) ont montré une excellente spécificité (100% et 96.8% respectivement) et une bonne sensibilité (76.5% et 88.2% respectivement), une spécificité correcte (82,3%) et une très faible sensibilité (23,5%) ont été obtenues pour la détection sérique de GM. Lexplication la plus logique à cette faible sensibilité réside probablement dans le caractère non-invasif de lANS dans lespèce canine. Dans une deuxième étude, nous avons tenté de déterminer le type de prélèvement nasal et la température dincubation les plus appropriés pour lobtention de résultats fiables lors de cultures fongiques de sécrétions nasales chez le chien. Trois types déchantillon nasal dinvasivité croissante (sécrétions nasales par écouvillon, biopsie de muqueuse nasale et placard fongique) et deux températures dincubation (air ambiant et 37°C) ont été comparés. Nos résultats ont montré que les placards fongiques étaient les échantillons de choix pour les cultures fongiques et quune incubation à 37°C améliorait également significativement la sensibilité de la culture fongique par rapport à la température ambiante (avec une sensibilité de 88% et une spécificité de 100%). Dans une troisième étude, nous avons investigué la valeur diagnostique des cultures fongiques à 37°C et de la détection de GM à partir de deux nouveaux types de prélèvement non-invasifs des sécrétions nasales (un nouvel écouvillon en nylon plus performant (le flocked swab (FSwab)) et du liquide de lavage nasal (LLN)) en comparaison avec les écouvillons traditionnels en coton (EC) et les placards fongiques. Malgré un nombre croissant de résultats de cultures positif avec lutilisation respective dEC, de FSwab et de LLN à partir des échantillons prélevés chez les chiens avec ANS, il ny avait pas de différence significative entre les types déchantillon. Même sil y avait significativement plus de GM au sein des LLN et des FSwab quau sein des EC si on tenait compte de lensemble des chiens (quil soient atteints dANS, de maladie non-fongiques ou indemnes de maladie respiratoire), lutilisation des ces deux nouvelles méthodes déchantillonnage na pas entrainé daugmentation significative des GM dans les sécrétions nasales des chiens atteints dANS par rapport à la quantité obtenue en utilisant les EC. Globalement, tous prélèvements confondus, ce test sest révélé peu sensible (41.7%) et modérément spécifique (77.4%) pour le diagnostic dANS et il ny a pas eu de différence de sensibilité en comparaison avec les cultures fongiques. Si le diagnostic de lANS représente un challenge pour le clinicien, son traitement lest tout autant. Aucune méthode thérapeutique nest à lheure actuelle totalement satisfaisante. En effet, soit la méthode nécessite un abord invasif des cavités nasales et/ou sinus frontaux, soit elle nécessite une anesthésie de longue durée. De plus, aucune des méthodes décrites nest efficace dans tous les cas. Le traitement idéal devrait donc être efficace, non-invasif et de courte durée. Cest pourquoi, dans une quatrième étude, nous avons testé lefficacité dun nouveau traitement topique rapide et non-invasif (dépôt sous endoscopie dune crème à base de bifonazole 1% (Canestene, Bayer)) comme traitement unique (DC) ou en combinaison avec un traitement classique dirrigation à lénilconazole 2% (DEC) chez des chiens atteints dANS. Il en a résulté que lajout du dépôt de bifonazole au protocole classique (DEC) offre une guérison dans 100% des cas après un voire deux traitements ; ces résultats sont comparables voire meilleurs que les résultats des autres études relatives au traitement topique de lANS chez le chien. Les résultats de cette étude ont montré également que le protocole DC nest efficace que lors dinfection fongique modérée et quil peut dès lors remplacer le protocole DEC lorsquil ne reste quun nombre réduit de placards fongiques après un traitement classique. Par ailleurs, la durée du protocole DC est nettement moins longue que celle du protocole DEC permettant de diminuer le temps danesthésie et donc le coût du traitement. Par contre lutilisation unique du protocole DC sur les cas dANS sévère semble peu efficace.

Clay minerals have historically played a consequential role in human health. While the beginnings were rooted in geophagy, a primitive act of consuming earth, the health-related uses of clay minerals have evolved and diversified over time. As excipients in pharmaceutical formulations, clay minerals can attribute novel properties onto intercalated compounds. Intercalating oxybenzone, a UV filter, within the interlamellar space of montmorillonite is desirable in order to minimize direct contact with skin. Intercalating resveratrol, a compound known for attributing beneficial effects onto human health, may be advantageous since this compound is susceptible to cis-trans isomerisation. The strategy of using alkylammonium–modified clay was undertaken and proved successful for the intercalation of oxybenzone. The field of biopolymer/layered silicate nanocomposites is heavily researched for use in a multitude of applications. Novel montmorillonite nanocomposites were prepared with neutral guar gum and cationic guar gum, using an environmentally friendly process and are fully characterized.

Clay minerals have historically played a consequential role in human health. While the beginnings were rooted in geophagy, a primitive act of consuming earth, the health-related uses of clay minerals have evolved and diversified over time. As excipients in pharmaceutical formulations, clay minerals can attribute novel properties onto intercalated compounds. Intercalating oxybenzone, a UV filter, within the interlamellar space of montmorillonite is desirable in order to minimize direct contact with skin. Intercalating resveratrol, a compound known for attributing beneficial effects onto human health, may be advantageous since this compound is susceptible to cis-trans isomerisation. The strategy of using alkylammonium–modified clay was undertaken and proved successful for the intercalation of oxybenzone. The field of biopolymer/layered silicate nanocomposites is heavily researched for use in a multitude of applications. Novel montmorillonite nanocomposites were prepared with neutral guar gum and cationic guar gum, using an environmentally friendly process and are fully characterized.

Clay minerals have historically played a consequential role in human health. While the beginnings were rooted in geophagy, a primitive act of consuming earth, the health-related uses of clay minerals have evolved and diversified over time. As excipients in pharmaceutical formulations, clay minerals can attribute novel properties onto intercalated compounds. Intercalating oxybenzone, a UV filter, within the interlamellar space of montmorillonite is desirable in order to minimize direct contact with skin. Intercalating resveratrol, a compound known for attributing beneficial effects onto human health, may be advantageous since this compound is susceptible to cis-trans isomerisation. The strategy of using alkylammonium–modified clay was undertaken and proved successful for the intercalation of oxybenzone. The field of biopolymer/layered silicate nanocomposites is heavily researched for use in a multitude of applications. Novel montmorillonite nanocomposites were prepared with neutral guar gum and cationic guar gum, using an environmentally friendly process and are fully characterized.

Estudos recentes têm sugerido que doença pulmonar obstrutiva crônica (DPOC) possa ser um fator de risco para aspergilose invasiva (AI), particularmente no contexto de ventilação mecânica e uso de esteróides. Neste trabalho, realizamos estudo de coorte prospectivo multicêntrico (2009-2010) em três unidades de terapia intensiva no Sul do Brasil. Foram incluídos no estudo pacientes com DPOC que apresentassem novo infiltrado pulmonar enquanto em ventilação mecânica e sob uso de corticosteróides. Para estes pacientes, foram realizados os seguintes testes, em amostras respiratórias (maioria aspirado traqueal): exame micológico direto, cultura quantitativa para fungos, pesquisa de antígeno galactomanana (GM) (Platelia Aspergillus) e PCR em tempo real para Aspergillus. O DNA das amostras respiratórias foi extraído utilizando-se o kit de extração MycXtra (Myconostica, UK), sendo a amplificação feita com dois kits comerciais de q-PCR: Aspergillus spp q-PCR Alert kit (Nanogen, Itália) e MycAssayTM Aspergillus kit (Myconostica, UK). Foi também obtido soro destes pacientes, onde foi testada GM, precipitinas para Aspergillus e IgE total. O estudo foi aprovado no comitê de ética dos dois hospitais. Foram incluídos no estudo 47 pacientes (40,4% do sexo masculino), sendo a idade média de 68,6 anos (±9,9). A maioria (72,8%) dos pacientes possuía DPOC grave (GOLD III/IV). A dosagem de esteróides (equivalentes de prednisona) variou de 100-4125 mg (mediana: 900 mg). Exame micológico (direto e cultivo) foi positivo para Aspergillus seção Fumigatti em apenas dois pacientes (4,2%). Outros fungos identificados foram Scedosporium apiospermum (n=1) e Histoplasma capsulatum (n=1). Precipitinas para Aspergillus foram positivas em três pacientes, com títulos baixos (<1:2). Os níveis de IgE variaram de 2 a >3000 UI/ml (mediana de 74 UI/ml). Em sua grande maioria, os índices de GM no soro foram <0,5, enquanto que nas amostras respiratórias, os índices de GM foram >0,5, >1,0 e >1,5 em 74,5%, 40,5% e 21,3%, respectivamente. PCR da Myconostica foi positivo em 10 pacientes, enquanto PCR Nanogen detectou apenas um paciente. A mortalidade geral foi de 53,2%. Este estudo prospectivo multicêntrico mostrou uma baixa incidência (4,2%) de AI em pacientes com DPOC. A determinação de GM mostrou altos índices nas amostras analisadas (50% com índices ópticos >1,3), possivelmente necessitando um maior ponto de corte para excluir resultados falso-positivos. A combinação de PCR e GM para o diagnóstico de AI em amostras respiratórias merece investigação adicional, devido à baixa sensibilidade dos métodos de cultivo observados nos estudos clínicos realizados. / Recent data have suggested that chronic obstructive pulmonary disease (COPD) may be an important risk factor for invasive aspergillosis (IA), particularly in the context of mechanical ventilation (MV) and therapy with corticosteroids. Here we present the results of a prospective multicentric study (2009-2010) conducted in three intensive care units (ICUs) in Southern Brazil. COPD patients on steroids showing a new lung infiltrate while on mechanical ventilation were included and the following tests were performed in respiratory samples (mostly tracheal aspirates): microscopy, quantitative fungal culture, galactomannan (GM) (Platelia Aspergillus EIA) and real-time PCR to detect Aspergillus DNA. DNA was extracted using MycXtra kit (Myconostica, UK) and amplification was performed using two q-PCR commercial kits: Aspergillus spp q-PCR Alert kit (Nanogen, Italy) and MycAssayTM Aspergillus kit (Myconostica, UK). Serum was also obtained and tested for Aspergillus precipitins, GM and total IgE levels. Ethical approval was obtained in each of the participant hospitals. A total of 47 patients were enrolled in the study (male 59.6%). Mean age was 68.6 years-old (± 9.9). Most patients had severe COPD (GOLD stages III/IV in 72.8%). Steroid dosage (prednisone equivalent) ranged from 100-4125 mg (median 900 mg). Microscopy and culture were positive for Aspergillus section Fumigatti in only 2 patients (4.2%). Other fungi included H. capsulatum (n=1) and S. apiospermum (n=1). Aspergillus precipitins were positive for three patients, at low titers (<1:2). IgE levels ranged from 2 to >3,000 IU/ml (median 74 IU/ml). All serum GM indexes were <0.5 and respiratory samples, GM indexes of >0.5, >1.0 and >1.5 were observed in 74.5%, 40.5%, and 21.3%, respectively. Myconostica PCR was positive in 10 patients, while Nanogen PCR detected only one patient. Overall mortality was 53.2%. This prospective multicenter study showed a low incidence (4.2%) of IA in critically ill patients with COPD. High optical indices were observed when GM was tested in respiratory samples (50% of the results showed indices of >1.3). Therefore, the test did not discriminate IA and a a higher cutoff would be needed to exclude false-positive results. The combination of PCR and GM for the diagnosis of IA in respiratory samples deserves further investigation due to the low diagnostic sensitivity of the classical mycology methods.

Estudos recentes têm sugerido que doença pulmonar obstrutiva crônica (DPOC) possa ser um fator de risco para aspergilose invasiva (AI), particularmente no contexto de ventilação mecânica e uso de esteróides. Neste trabalho, realizamos estudo de coorte prospectivo multicêntrico (2009-2010) em três unidades de terapia intensiva no Sul do Brasil. Foram incluídos no estudo pacientes com DPOC que apresentassem novo infiltrado pulmonar enquanto em ventilação mecânica e sob uso de corticosteróides. Para estes pacientes, foram realizados os seguintes testes, em amostras respiratórias (maioria aspirado traqueal): exame micológico direto, cultura quantitativa para fungos, pesquisa de antígeno galactomanana (GM) (Platelia Aspergillus) e PCR em tempo real para Aspergillus. O DNA das amostras respiratórias foi extraído utilizando-se o kit de extração MycXtra (Myconostica, UK), sendo a amplificação feita com dois kits comerciais de q-PCR: Aspergillus spp q-PCR Alert kit (Nanogen, Itália) e MycAssayTM Aspergillus kit (Myconostica, UK). Foi também obtido soro destes pacientes, onde foi testada GM, precipitinas para Aspergillus e IgE total. O estudo foi aprovado no comitê de ética dos dois hospitais. Foram incluídos no estudo 47 pacientes (40,4% do sexo masculino), sendo a idade média de 68,6 anos (±9,9). A maioria (72,8%) dos pacientes possuía DPOC grave (GOLD III/IV). A dosagem de esteróides (equivalentes de prednisona) variou de 100-4125 mg (mediana: 900 mg). Exame micológico (direto e cultivo) foi positivo para Aspergillus seção Fumigatti em apenas dois pacientes (4,2%). Outros fungos identificados foram Scedosporium apiospermum (n=1) e Histoplasma capsulatum (n=1). Precipitinas para Aspergillus foram positivas em três pacientes, com títulos baixos (<1:2). Os níveis de IgE variaram de 2 a >3000 UI/ml (mediana de 74 UI/ml). Em sua grande maioria, os índices de GM no soro foram <0,5, enquanto que nas amostras respiratórias, os índices de GM foram >0,5, >1,0 e >1,5 em 74,5%, 40,5% e 21,3%, respectivamente. PCR da Myconostica foi positivo em 10 pacientes, enquanto PCR Nanogen detectou apenas um paciente. A mortalidade geral foi de 53,2%. Este estudo prospectivo multicêntrico mostrou uma baixa incidência (4,2%) de AI em pacientes com DPOC. A determinação de GM mostrou altos índices nas amostras analisadas (50% com índices ópticos >1,3), possivelmente necessitando um maior ponto de corte para excluir resultados falso-positivos. A combinação de PCR e GM para o diagnóstico de AI em amostras respiratórias merece investigação adicional, devido à baixa sensibilidade dos métodos de cultivo observados nos estudos clínicos realizados. / Recent data have suggested that chronic obstructive pulmonary disease (COPD) may be an important risk factor for invasive aspergillosis (IA), particularly in the context of mechanical ventilation (MV) and therapy with corticosteroids. Here we present the results of a prospective multicentric study (2009-2010) conducted in three intensive care units (ICUs) in Southern Brazil. COPD patients on steroids showing a new lung infiltrate while on mechanical ventilation were included and the following tests were performed in respiratory samples (mostly tracheal aspirates): microscopy, quantitative fungal culture, galactomannan (GM) (Platelia Aspergillus EIA) and real-time PCR to detect Aspergillus DNA. DNA was extracted using MycXtra kit (Myconostica, UK) and amplification was performed using two q-PCR commercial kits: Aspergillus spp q-PCR Alert kit (Nanogen, Italy) and MycAssayTM Aspergillus kit (Myconostica, UK). Serum was also obtained and tested for Aspergillus precipitins, GM and total IgE levels. Ethical approval was obtained in each of the participant hospitals. A total of 47 patients were enrolled in the study (male 59.6%). Mean age was 68.6 years-old (± 9.9). Most patients had severe COPD (GOLD stages III/IV in 72.8%). Steroid dosage (prednisone equivalent) ranged from 100-4125 mg (median 900 mg). Microscopy and culture were positive for Aspergillus section Fumigatti in only 2 patients (4.2%). Other fungi included H. capsulatum (n=1) and S. apiospermum (n=1). Aspergillus precipitins were positive for three patients, at low titers (<1:2). IgE levels ranged from 2 to >3,000 IU/ml (median 74 IU/ml). All serum GM indexes were <0.5 and respiratory samples, GM indexes of >0.5, >1.0 and >1.5 were observed in 74.5%, 40.5%, and 21.3%, respectively. Myconostica PCR was positive in 10 patients, while Nanogen PCR detected only one patient. Overall mortality was 53.2%. This prospective multicenter study showed a low incidence (4.2%) of IA in critically ill patients with COPD. High optical indices were observed when GM was tested in respiratory samples (50% of the results showed indices of >1.3). Therefore, the test did not discriminate IA and a a higher cutoff would be needed to exclude false-positive results. The combination of PCR and GM for the diagnosis of IA in respiratory samples deserves further investigation due to the low diagnostic sensitivity of the classical mycology methods.

CoordenaÃÃo de AperfeÃoamento de Pessoal de NÃvel Superior / Este trabalho teve como objetivo a preparaÃÃo de nanopartÃculas de derivados da galactomanana. O polissacarÃdeo (galactomanana) foi extraÃdo de sementes da Dimorphandra gardneriana com rendimento de 29Â3% e razÃo manose/ galactose (M:G) de 1,93Â0,02. A galactomanana foi modificada por reaÃÃo de sulfataÃÃo (FDS), oxidaÃÃo (FDO) e por acilaÃÃo com anidrido acÃtico (FDAc) e com anidrido propiÃnico (FDPr). Os derivados foram utilizados no desenvolvimento de nanopartÃculas por interaÃÃo com a quitosana via complexaÃÃo polieletrolÃtica, por formaÃÃo de base de Schiff, e por auto-organizaÃÃo. Os derivados da galactomanana foram caracterizados por espectroscopia na regiÃo do infravermelho (IV), por ressonÃncia magnÃtica nuclear, por anÃlise elementar, por cromatografia de permeaÃÃo em gel e por viscosidade intrÃnseca. O grau de substituiÃÃo para reaÃÃo de sulfataÃÃo foram 0,32 e 0,42. A razÃo de unidades glicosÃdicas/unidades oxidadas para os derivados oxidados foram 10:2; 10:4 e 10:8. Os derivados hidrofÃbicos foram confirmados pelo aparecimento de uma nova absorÃÃo em 1740 cm-1 no IV, atribuÃda ao estiramento C=O e a insolubilidade em Ãgua. NanopartÃculas com perfil unimodal foram obtidas nas trÃs rotas estudadas. As partÃculas de quitosana e galactomanana sulfatada foram obtidas por complexaÃÃo polieletrolÃtica com diÃmetros variando de 10Â6 a 377Â29 nm, Ãndice de polidispersividade de 0,11Â0,02 a 0,5Â0,1 e potencial zeta de â28Â2 a 59Â4 mV. Para as de quitosana e galactomanana oxidada obtidas via base de Schiff com diÃmetros variando de 8Â2 a 20Â9 nm, Ãndice de polidispersividade de 0,36Â0,04 a 0,57Â0,08 e potencial zeta de 0,4Â0,5 a 14Â1 mV. PartÃculas de galactomanana modificadas hidrofobicamente foram obtidas via auto-organizaÃÃo com diÃmetros variando de 35,3Â0,6 a 213Â28 nm, Ãndice de polidispersividade de 0,093Â0,003 a 0,9Â0,1 e potencial zeta de â26,3Â0,9 a â3Â2 mV. O diÃmetro, potencial zeta e Ãndice de poidispersividade sÃo influenciados pela metodologia e derivado utilizado, mas para todos os derivados da galactomanana obteve-se, em pelo menos uma condiÃÃo, nanopartÃculas compatÃveis para um potencial uso como carreadores de fÃrmacos. Fatores como: grau de modificaÃÃo, razÃo, ordem de adiÃÃo e concentraÃÃo dos polissacarÃdeos, pH e a adiÃÃo de fÃrmaco influenciaram no tamanho, Ãndice de polidispersividade, potencial zeta e estabilidade em soluÃÃo das partÃculas. / This work aimed the preparation of nanoparticles galactomannan derivatives. The polysaccharide (galactomannan) was from seeds of Dimorphandra gardneriana with yield of 29Â3% and mannose/galactose ratio (M:G) of 1.93Â0.02. The galactomannan was modified by sulfation raction (FDS), oxidation (FDO) and by acylation with acetic anhydride (FDAc) and proponic (FDPr) and derivates were used in the developmentof nanoparticles by interaction with chitosan by polyelectrolyte complexes, Schiff base formation and by self-assembled. Galactomannan derivatives were characterized by infrared spectroscopy (IR), nuclear magnetic resonance, elemental analysis, gel permeation chromatography and intrinsic viscosity. The degree of substitution for sulfataion reaction were 0.32 and 0.42; the ratio of glycosidic/oxidized units in oxidized derivatives was 10:2; 10:4 and 10:8. The hydrophobic derivatives were confirmed by the appearance of new absorption at 1740 cm-1 in IR, assigned to C=O stretching and water insolubility. Nanoparticles with unimodal profile were obtained on three routes studied. The particles of chitosan and sulfated galactomannan were obtained by complexation polyelectrolytic with diameters ranging from 10Â6 to 377Â29 nm, polydispersity index from 0.11Â0.02 to 0.5Â0.1 and zeta potential of â28Â2 to 59Â4 mV. For chitosan and oxidized galactomannan nanoparticles, obtained by Schiff base, diameters ranging from 8Â2 to 20Â9 nm, polydispersity index from 0.36Â0.04 to 0.57Â0.08 and zeta potential of 0.4Â0.5 to 14Â1 mV were obatined. Hydrophobically modified galactomannan particles were obtained by self-assembled with diameters ranging from 35.3Â0.6 to 213Â28 nm, polydispersity index from 0.093Â0.003 to 0.9Â0.1 and zeta potential of â26.3Â0.9 to â3Â2 mV. The diameter, zeta potential and polydispersity index are influenced by the methodology and derived used, but for all derivatives of galactomannan at least one condition shows nanoparticles compatible for potential use as drug carriers. Factors such as degree of modification, ratio, order of addition and concentration of polysaccharides, pH and addition of drug have influence on size, polydispersity index, zeta potential and solution stability of the particles.