The objective of this research was to evaluate if chitosan and another food-grade compound can be incorporated into food-grade coatings in conjunction with propylene glycol and polysaccharides to control Tyrophagus putrescentiae growth without affecting sensory attributes, water activity, and moisture content of dry-cured hams. Each coating solution was coated on ham cubes (2.5 × 2.5 × 2.5 cm3, n = 5/treatment) or infused in ham nets, and dry-cured ham cubes were wrapped in the ham nets prior to inoculation with 20 mixed-sex adult mites. Randomized complete block designs with two replications were utilized to evaluate the efficacy of treatments at controlling mite growth on dry-cured ham cubes. When CH was mixed with XG and infused into a net, fewer mites (15.7 and 21.0 mites) were on ham cubes (P < 0.05) in comparison to the control (211.2 mites). Results indicated that CH contributed to controlling mite growth since 1% XG alone did not control mite growth. Difference from control test results indicated that no sensory differences existed (NS) between CH-treated and control ham slices. The addition of a food-grade compound (SP or 24P) to the XG coatings enhanced mite control efficacy. When 1% SP was added to 1% XG, it effectively controlled mite growth in both coating and netting treatments. Increasing 1% SP to 2% SP did not significantly (NS) control mite growth in the coating solution but did control mite growth when infused in the net. Both coating and netting treatments with 2% 24P + 1% XG controlled mite growth. When 24P was the only ingredient in the treatment, both 1% and 2% 24P controlled mites when infused in the net. The addition of SP did not (NS) impact the sensory attributes of the dry-cured ham. The 2% 24P + 1% XG treatment differed in moistness (P < 0.05) when compared to the blind control, but the rating was still less than 2 (moderately different). Overall, results from these studies indicate that chitosan can potentially be added in coatings or ham nets as tools to control mites in an integrated pest management program for dry-cured hams.

This diploma thesis was focused on the study of lysozyme and lysozyme-like proteins, either of similar function (antibacterial) or molecular weight (14 - 17 kDa), of synanthropic acaroid mites. In general, animals utilize lysozymes for defensive (antimicrobial) or digestive purposes but also as a digestive enzyme. Some chitinases or other enzymes that act similarly to lysozyme can be utilized for similar purposes. Chitinases belong to house dust mite allergens. One of major mite are historically named lysozyme-like proteins which name relates to their size similar to lysozyme. Bacteriolytic activity has also 14.5 kDa (UniprotKB Q8MWR6) protein. The species selected for the study were domestic mites Dermatophagoides farinae, D. pteronyssinus and Lepidoglyphus destructor. Presence of lysozyme was detected by direct detection with polyclonal antibody using immunohistochemistry and dot blots. Immunohistochemistry proved presence of lysozyme epitopes in the feces of D. farinae, D pteronyssinus a L. destructor. Dot blot analysis demonstrated the presence of imunoreactivity of antibody in spent growth medium extracts (SGME) of all three species. This implies that lysozyme is synthesized in the midgut. The presence of lysozyme and lysozyme-like proteins was proved using 2D electrophoresis and MALDI TOF/TOF...

Die Prävalenz allergischer Erkrankungen und des Asthma bronchiale haben bei Kindern und Jugendlichen in den letzten 20 Jahren deutlich zugenommen. Ursächlich scheinen dafür eher veränderte Umweltfaktoren als genetische Mutationen verantwortlich zu sein. Faktoren des Lebensstils, die sich in den letzten 50 Jahren gewandelt haben, wie z. B. Abnahme von schweren Infektionserkrankungen durch Verbesserung der Hygiene und Entwicklung von Antibiotika, Adipositas und Bewegungsmangel, Abnahme der Familiengröße, veränderte Ernährungsgewohnheiten und eine Zunahme der Innenraumallergen-Konzentration durch bauliche Veränderungen kommen als potentielle Risikofaktoren in Betracht. In einer prospektiven Geburtskohortenstudie (Multizentrische Allergiestudie MAS90) untersuchten wir relevante Expositionsfaktoren wie z. B. Milben- und Katzenallergene im Hausstaub, frühkindliche Infekte und vieles mehr in Hinblick auf die Entstehung des kindlichen Asthma bronchiale. Von den ursprünglich im Jahr 1990 rekrutierten 1314 Neugeborenen lagen uns zum Zeitpunkt 7. Geburtstag von 939 Kindern komplette Informationen vor. Die regelmäßigen Untersuchungen umfassten jährliche Interviews und Blutentnahmen zur Bestimmung von spezifischem Serum-IgE gegen Nahrungsmittel- und Inhalationsallergene und die Quantifizierung von Hausstaubmilben (Dermatophagoides)- und Katzenallergenen im Teppich- und Matratzenstaub an mehreren Zeitpunkten. Im Alter von 7 Jahren wurde bei 645 Kindern eine Lungenfunktionsuntersuchung sowie die Messung der bronchialen Überempfindlichkeit mit Hilfe der Histamin-Provokation gemessen. Bei 10% (94 von 938) der 7-Jährigen trat eine pfeifende Atmung im Beobachtungszeitraum der letzten 12 Monate vor der Untersuchung auf, 6,1% (57 von 939) der Kinder hatten die Arztdiagnose "Asthma bronchiale". Eine Sensibilisierung gegen Innenraumallergene war mit Asthma bronchiale und erhöhter bronchialer Empfindlichkeit assoziiert, es gab jedoch keine Beziehung zwischen der Asthmaprävalenz und der Höhe der Milben- oder Katzenallergenexposition im Hausstaub. Unsere Ergebnisse geben keinen Anhalt für eine kausale Beziehung zwischen Innenraumallergenen und der Entwicklung des kindlichen Asthma bronchiale. Vielmehr scheinen allergische Immunantwort und die Manifestation eines Asthma bronchiale von unterschiedlichen genetischen und Umweltfaktoren beeinflusst zu werden. Die Vermeidung von Innenraumallergenen im Rahmen von Sekundär- und Tertiärprävention scheint weiterhin sinnvoll zu sein, jedoch scheint der Erfolg von Allergenreduktion im häuslichen Milieu hinsichtlich der Senkung der Asthmaprävalenz im Kindesalter zweifelhaft. Beim allergisches Asthma bronchiale als Th2-Erkrankung des Immunsystems scheinen insbesondere immunmodulatorische Faktoren wie frühkindliche Infektionen bzw. hoher Endotoxinexposition oder anderen Faktoren, die Gen-Umweltinteraktion und Suszeptibilität des Individuums beeinflussen, von Bedeutung zu sein. / Epidemiological surveys indicate that there has been a notable increase in the prevalence of both asthma and other allergic symptoms in children and young adults. Since it seems unlikely that genetic factors contribute to the rising trend, environmental factors might play a major part in the development of childhood asthma. Several potential determinants have been proposed, such as lack of severe and repeated infections, obesity and lack of physical exercise, decreased family size, changing dietary habits, and increase of indoor allergen exposure. In a prospective birth-cohort study, we assessed the relevance of different exposures like mite and cat allergen exposure and early infectious diseases for the development of childhood asthma up to the age of 7 years. Of 1314 newborn infants enrolled in five German cities in 1990, follow-up data at age 7 years were available for 939 children. Assessments included repeated measurements of specific IgE to food and inhalant allergens, measurement of indoor allergen exposure at 6 months, 18 months, and 3 years of age, and yearly interviews by a paediatrician. At age 7 years, pulmonary function was tested and bronchial hyper-responsiveness was measured in 645 children. At age 7, the prevalence of wheezing in the past 12 months was 10 % (94 of 938), and 6.1 % (57 of 939) parents reported a doctor`s diagnosis of asthma in their children. Sensitization to indoor allergens was associated with asthma, wheeze, and increased bronchial hyper-responsiveness. However, no relation between early indoor allergen exposure and the prevalence of asthma, wheeze, and bronchial-hyperresponsiveness was seen. Our data do not support the hypothesis that exposure to environmental allergen causes asthma in childhood, but rather the induction of specific IgE responses and the development of childhood asthma are determined by independent factors. Indoor allergen avoidance is recommendable as first line of treatment in secondary and tertiary prevention, however, conclusions should be drawn with caution about the possible effect of primary prevention measures. As allergic asthma seems to be a Th2-disease, immunomodulating factors like early childhood infections and LPS-exposure or other factors influencing gene-environment interaction and individual susceptibility seem to be relevant for the development of childhood asthma.

Submitted by PPG Pediatria e Sa?de da Crian?a (pediatria-pg@pucrs.br) on 2018-02-16T19:15:31Z No. of bitstreams: 1 Tese final 2018 - artigo publicado.pdf: 1647298 bytes, checksum: 03a39af4be2d469330c9618fb70f9cdd (MD5) / Approved for entry into archive by Caroline Xavier (caroline.xavier@pucrs.br) on 2018-02-23T16:28:05Z (GMT) No. of bitstreams: 1 Tese final 2018 - artigo publicado.pdf: 1647298 bytes, checksum: 03a39af4be2d469330c9618fb70f9cdd (MD5) / Made available in DSpace on 2018-02-23T16:35:09Z (GMT). No. of bitstreams: 1 Tese final 2018 - artigo publicado.pdf: 1647298 bytes, checksum: 03a39af4be2d469330c9618fb70f9cdd (MD5) Previous issue date: 2017-07-12 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Background: Asthma is a chronic disease of the airways, characterized by bronchial inflammation and hyperresponsiveness, which affects approximately 300 million people around the world. The increase in the prevalence of asthma in recent years has been associated with an increase in vitamin D deficiency. About 1 billion people in the world have insufficient levels of vitamin D due to many factors such as reduced outdoor activities, sunscreens use and a diet low in vitamin D. In addition, many studies suggest that vitamin D deficiency has a direct effect on lung function, leading to changes in the structure of the airways and in the inflammatory process. Objectives: To evaluate the effect of vitamin D deficiency at different life stages in a murine model of allergic airways disease house dust mite (HDM) induced on inflammation and lung function. Methods: Female BALB / c mice were placed in a diet replete or deficient in vitamin D at three-week old. At 8 weeks, females were mated with males on a diet replete in vitamin D. At birth, pups were cross-fostered to assess the effects of vitamin D deficiency at different stages of life, in utero (Vit D -/+), postnatal (Vit D +/-) and whole-life (Vit D - / -) compared to the control group whole-life vitamin D replete (Vit D + / +). At 8 weeks of age, mice of both sexes were challenged for 10 consecutive days intranasally with either HDM extract or saline solution after mild anesthesia. The animals were anesthetized for lung function test and then submitted to euthanasia for bronchoalveolar lavage (BAL) and lung tissue removal 24 hours after the last intranasal challenge. The total BAL cell count and collagen quantification of lung tissue homogenized were evaluated. Results: Vitamin D deficiency did not affect HDM-induced inflammation, which was characterized by BAL eosinophilia. Vitamin D deficiency at any life stage (in utero, postnatal and all life) caused impairment of lung function, increased tissue damping and tissue elastance, being particularly observed in females. On the other hand, the asthma HDM-induced decreased airway distensibility, but only in females and vitamin D do not altered this response. Conclusion: Our results suggest that vitamin D and HDM have different mechanisms that influence in the development of allergic lung disease and furthermore the effects appear to be sex-specific. / Introdu??o: A asma ? uma doen?a cr?nica das vias a?reas, caracterizada por inflama??o e hiperresponsividade br?nquica, que atinge aproximadamente 300 milh?es de pessoas ao redor do mundo. O aumento da preval?ncia da asma nos ?ltimos anos tem sido associado ao aumento da defici?ncia de vitamina D. Cerca de 1 bilh?o de pessoas no mundo apresenta n?veis insuficientes de vitamina D em fun??o de diversos fatores, tais como a redu??o de atividades ao ar livre, uso de protetor solar e dieta pobre em vitamina D. Al?m disso, estudos sugerem que a defici?ncia de vitamina D possui um efeito direto na fun??o pulmonar, causando altera??es na estrutura das vias a?reas e no processo inflamat?rio. Objetivo: Avaliar o efeito da defici?ncia de vitamina D em diferentes est?gios da vida de camundongos com asma induzida por ?caro domiciliar (house dust mite; HDM) sobre a inflama??o e fun??o pulmonar. M?todos: Camundongos BALB/c f?meas receberam dieta rica ou deficiente em vitamina D a partir da terceira semana de vida. Com 8 semanas de vida, as f?meas foram acasaladas com machos em dieta rica em vitamina D. Ao nascimento foi realizado o cross-fostering (ado??o cruzada) com a prole para que fosse poss?vel avaliar o efeito da defici?ncia de vitamina D em diferentes est?gios da vida, in utero (Vit D -/+), p?s-natal (Vit D +/-) e durante toda a vida (Vit D -/-), em compara??o ao grupo controle, que recebeu dieta rica em vitamina D durante toda a vida (Vit D +/+). Com 8 semanas de vida, camundongos de ambos os sexos foram desafiados por 10 dias consecutivos por via intranasal, com extrato de HDM ou apenas solu??o salina. Os animais foram anestesiados para a realiza??o do teste de fun??o pulmonar e ent?o submetidos a eutan?sia para a realiza??o do lavado broncoalveolar (LBA) e retirada do tecido pulmonar, 24 horas ap?s o ?ltimo desafio intranasal. Foi avaliada a contagem total de c?lulas do LBA e quantifica??o de col?geno no homogeneizado de tecido pulmonar. Resultados: A defici?ncia de vitamina D n?o afetou a inflama??o induzida por HDM, que foi caracterizada por eosinofilia no LBA. A defici?ncia de vitamina D em qualquer fase da vida dos camundongos (in utero, p?s-natal e durante toda a vida) causou uma piora na fun??o pulmonar, aumentando o tissue damping e tissue elastance, sendo observado particularmente em f?meas. Por outro lado, a asma induzida por HDM diminuiu a distensibilidade das vias a?reas apenas em f?meas e a vitamina D n?o alterou essa resposta. Conclus?o: Nossos resultados sugerem que a vitamina D e HDM possuem diferentes mecanismos que influenciam no desenvolvimento da doen?a pulmonar al?rgica e, al?m disso, os efeitos parecem ser dependentes do sexo.

O sul de Minas Gerais é a principal região produtora de morangos (Fragaria x ananassa Duch.) no Brasil. O ácaro-rajado, Tetranychus urticae Koch (Acari: Tetranychidae), é considerado uma das principais pragas do morangueiro no Brasil e em vários países. Uma prática importante na cultura do morango refere-se à cobertura de solo, usualmente realizada com filme de polietileno, porém além de ser caro gera resíduos muito persistentes no ambiente. Objetivou-se com este trabalho comparar a resistência de oito genótipos de morango (\'Albion\', \'IAC Guarani\', \'IAC Princesa Isabel\', \'Oso Grande\', IAC T-0104, IAC 12, IAC 4 e IAC 1.13) ao ácaro-rajado; determinar o efeito da cobertura de solo (com a polpa de café desidratada, conhecida como palha de café) nas pragas/ patógenos e ácaros predadores desta cultura; comparar a ocorrência de pragas, patógenos e inimigos naturais entre um cultivo orgânico e outro convencional de morango. Os resultados obtidos indicaram que \'IAC Princesa Isabel\', IAC T-0104 e IAC 12 são resistentes, IAC 1.13 e IAC 4 apresentam resistência moderada e \'IAC Guarani\', \'Oso Grande\' e \'Albion\' são suscetíveis ao ácaro-rajado. \'Oso Grande\' e \'Albion\' são bastante utilizadas pelos produtores do sul de Minas Gerais. O uso de palha de café aumentou o número de predadores edáficos, tanto no solo dos canteiros (campo) e em vasos (laboratório) como em folíolos de morangueiro. O Gamasina Proctolaelaps pygmaeus (Müller) (Melicharidae) foi visto sobre morangueiros, principalmente no período noturno. Maior número de ácaro-rajado e maior severidade de doenças foram observados em plantas cultivadas em solo coberto com polietileno. Além disso, foi maior o nível de infecção de ácaro-rajado pelo fungo Neozygites floridana (Weiser e Muma) em plantas cultivadas em solo coberto com palha de café. Não foi observada diferença significativa entre a produtividade da cultura em solo coberto com polietileno e em solo coberto com palha seca de café. Os resultados da comparação da ocorrência de ártropodes e patógenos entre sistema orgânico e convencional mostraram menor ocorrência de ácaro-rajado e predadores em cultivo de morangueiro convencional, porém ocorrência considerável de tripes, mosca-branca e mofo cinzento. A incidência de dendrofoma e mancha de pestalotia foi a mesma nos dois sistemas de cultivos. Os resultados sugerem a conveniência de se continuar o processo de desenvolvimento dos genótipos que se mostraram menos afetados pelo ácaro-rajado, para que estes possam no futuro ser utilizados pelos produtores, e a condução de estudos complementares, que avaliem o desempenho de cultivos em sistema orgânico que incorporem o uso da palha de café para a cobertura do solo. / The southern of Minas Gerais is the main region producing strawberries (Fragaria x ananassa Duch.) in Brazil. The two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), is considered one of the main strawberry pests in Brazil and in several countries. An important practice in strawberry cultivation refers to soil cover, which is usually made with polyethylene film, but it is expensive and generates very persistent residues in the environment. The objective of this study was to compare the resistance of eight strawberry genotypes (\'Albion\', \'IAC Guarani\', \'IAC Princesa Isabel\', \'Oso Grande\', IAC T-0104, IAC 12, IAC 4 and IAC 1.13); to determine the effect of soil cover (with dehydrated coffee pulp, known as coffee straw) on the pests mites/ pathogens and predatory mites of this crop; to compare the occurrence of pests, pathogens and natural enemies between an organic and a conventional strawberry crop. The results indicated that \'IAC Princesa Isabel\', IAC T-0104 and IAC 12 are resistant, IAC 4 and IAC 1.13 are moderately resistant, and \'IAC Guarani\', \'Oso Grande\' and \'Albion\' are susceptible to the two-spotted spider mite. \'Oso Grande\' and \'Albion\' are widely used by producers in the southern of Minas Gerais. The use of coffee straw increased the number of edaphic predators, both in the soil beds (field) and in pots (laboratory) as well as in strawberry leaflets. The Gamasina Proctolaelaps pygmaeus (Müller) (Melicharidae) was seen on strawberry leaftlets, mainly in the nocturnal period. Higher numbers of the two-spotted spider mites and disease severity were observed in plants grown on soil covered with polyethylene. In addition, the level of the two-spotted spider mite infected by the fungi Neozygites floridana (Weiser and Muma) was higher in plants cultivated in soil covered with coffee straw. No significant difference was observed between yield in soil covered with polyethylene and in soil covered with dry coffee straw. The results of the comparison of the occurrence of arthropods and pathogens between the organic and conventional systems showed fewer occurrences of the two-spotted spider mites and predators in conventional strawberry cultivation, but considerable occurrence of thrips, whitefly and gray mold. The incidence of dendrophoma and leaf spots caused by pestalotia was the same in both cropping systems. The results suggest the convenience to continue the development process of the genotypes that were less affected by the two-spotted spider mite, which could be used in the future by producers, and the conduction of complementary studies to evaluate the performance of crops in organic system that incorporate the use of coffee straw as soil coverage.

Submitted by (edna.saturno@ufrpe.br) on 2016-12-19T15:11:21Z No. of bitstreams: 1 Maria Virginia Alves Xavier.pdf: 938724 bytes, checksum: 3969de87ac989fbf6102e3ade913c13d (MD5) / Made available in DSpace on 2016-12-19T15:11:21Z (GMT). No. of bitstreams: 1 Maria Virginia Alves Xavier.pdf: 938724 bytes, checksum: 3969de87ac989fbf6102e3ade913c13d (MD5) Previous issue date: 2014-02-24 / Biodiesel production has received enough encouragement and among the crops with potential applicability to this highlights the physic nut (Jatropha curcas L. ) , which has been identified as one of the most promising crops for biodiesel and inclusion in family production chain , which has opened up broad prospects for growth in the areas of planting this crop in semi-arid northeast . Several factors limit the productivity of Jatropha, with emphasis on the attack by pests such as spider mite Tetranychus bastosi Tuttle , Baker & Sales (Acari : Tetranychidae) . Among the methods currently used for alternative pest control is gaining prominence employment of plant extracts due to the presence of secondary metabolites present in some plants and their low toxicity to the environment and to humans . In the present work we evaluated the potential of plant extracts Myracrodruon urundeuva All Br ( Anacardiaceae ) , Croton blanchetianus Baill (Euphorbiaceae) and Ziziphus joazeiro Mart . (Rhamnaceae ) on the mite T. bastosi associated with the culture of Jatropha. We evaluated the effect of the extract of M. urundeuva at different doses (0 , 5 % , 10 % , 15 % , 20 % and 25 % ) on the biology and fertility life table T. bastosi . It was observed that the extract of M. urundeuva prolonged cycle of development of the mite and with increasing dosages, prolonged the longevity of T. bastosi . With respect to the life and fertility table, it was found that there was no effect of the extract for the parameters, except for the survival and viability of eggs, where there was a lower progeny survival at a dose of 10% and a lower egg viability in strengths of 15 % and 20 %. Toxicity and repellency of the extracts of M. urundeuva , C. blanchetianus and Z. joazeiro was also evaluated . Generally extracts showed toxic effect on adults of T. bastosi the concentrations tested. The extract of Z. joazeiro showed the highest mortality rates (90 %) mean mortality of individuals. With regard to the repellency of these extracts, all treatments were shown repellents for females of Tetranychus bastosi classified as repellent treatment, except for the 5% dose of extract of M. unrundeuva . / A produção de biodiesel vem recebendo bastante incentivo e dentre as culturas agrícolas com potencial para esta aplicabilidade destaca-se o pinhão-manso (Jatropha curcas L.), que vem sendo apontada como uma das mais promissoras para o biodiesel e inserção na cadeia produtiva familiar. Isso tem aberto amplas perspectivas para o crescimento das áreas de plantio desta cultura no semiárido nordestino. Diversos fatores limitam a produtividade do pinhão-manso, havendo destaque para o ataque por pragas, como o ácaro Tetranychus bastosi Tuttle, Baker & Sales (Acari: Tetranychidae), o qual foi registrado recentemente em Pernambuco. Dentre os métodos utilizados atualmente para o controle alternativo de pragas vem ganhando destaque o emprego de extratos vegetais, devido à presença metábólitos secundários presentes em algumas plantas e à sua baixa toxicidade para o meio ambiente e para o homem. No presente trabalho avaliou-se o potencial de extratos vegetais de Myracrodruon urundeuva Fr. All (Anacardiaceae), Croton blanchetianus Baill (Euphorbiaceae) e Ziziphus joazeiro Mart. (Rhamnaceae,) sobre o ácaro T. bastosi associado à cultura do pinhão- manso. Avaliou-se o efeito do extrato de M. urundeuva, em diferentes dosagens (0, 5%, 10%, 15%, 20% e 25%) sobre a biologia e a tabela de vida de fertilidade de T. bastosi. Observou-se que o extrato de M. urundeuva prolongou o ciclo de desenvolvimento deste ácaro e com o aumento das dosagens, houve um prolongamento da longevidade de T. bastosi. No que se refere à tabela de vida e fertilidade, verificou-se que não houve efeito do extrato para os parâmetros analisados, exceto para a sobrevivência e viabilidade de ovos, onde observou-se uma menor sobrevivência da progênie na dose de 10% e uma menor viabilidade de ovos nas dosagens de 15% e 20%. Também foi avaliada a toxicidade e repelência dos extratos de M. urundeuva, C. blanchetianus e Z. joazeiro sobre T. bastosi. De uma forma geral os extratos demonstraram efeito tóxico para adultos de T. bastosi nas concentrações testadas. O extrato de Z. joazeiro apresentou as maiores taxas de mortalidade (90% de mortalidade média dos indivíduos). No que se refere à repelência destes extratos, todos os tratamentos se mostraram repelentes para fêmeas de Tetranychus bastosi, classificados como tratamentos repelentes, exceto para a dosagem de 5% do extrato de M. unrundeuva.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A asma é um distúrbio crônico pulmonar caracterizado por inflamação, obstrução e remodelamento brônquico, levando a sintomas como sibilo, tosse e falta de ar. A terapia antiasmática consiste em corticosteroides inalados e agonistas &#946;2 de curta ou longa duração. O tratamento é limitado por efeitos colaterais e refratariedade de alguns pacientes, justificando a necessidade de novas terapias. Estudos demonstram que a 15-deoxy-delta- 12,14-prostaglandina J2 (15d-PGJ2), um ligante endógeno de receptores ativados por proliferadores de peroxissomos do tipo gama (PPAR-&#947;), é capaz de reduzir a expressão de citocinas pró-inflamatórias, o que pode resultar em benefícios no tratamento de doenças com esse perfil. O objetivo deste estudo foi avaliar o potencial anti-inflamatório e antiasmático da 15d-PGJ2 em modelos experimentais de asma. Camundongos A/J machos foram sensibilizados nos dias 0 e 7 através de injeção subcutânea (s.c.), contendo ovoalbumina (OVA) e Al(OH)3, e desafiados com 4 instilações intranasais (i.n.) de OVA em intervalos semanais. O tratamento com 15d-PGJ2 (30 e 100 &#61549;g/Kg, s.c.) foi realizado 30 min antes dos desafios a partir da terceira provocação antigênica. Em outro modelo, camundongos A/J foram desafiados intranasalmente com extrato de ácaro 3 vezes por semana durante 3 semanas. As administrações de 15d-PGJ2 (30, 70 e 100 g/Kg, s.c. e 0,65; 1,5 e 2,3 g/animal, i.n.) foram realizadas a partir da 3 semana, 30 min antes dos desafios. As análises ocorreram 24 h após o último desafio. Nossos resultados mostraram que, em camundongos previamente sensibilizados e desafiados com OVA, a administração de 15d-PGJ2 limitou significativamente o influxo peribrônquico de eosinófilos e neutrófilos, bem como a produção de muco por células caliciformes e fibrose sub-epitelial, além da hiperreatividade das vias aéreas e produção de IL-5. A redução do epitélio brônquico e das citocinas IL-13 e TNF-&#945; foram observadas somente na maior dose administrada. No modelo HDM a inflamação e o remodelamento foram atenuados em todas as doses administradas do composto, enquanto que a hiperresponssividade brônquica foi inibida apenas nas doses de 70 e 100 &#956;g/Kg (via sistêmica) e na dose intermediária dada topicamente (1,5 &#956;g/animal, i.n.). Os níveis de citocinas foram atenuados pelo tratamento subcutâneo, porém somente os níveis de IL-17, eotaxina-1 e TNF-&#945; foram inibidos com a dose intranasal de 0,65 g/animal. O aumento da expressão de NF-&#954;B, induzido por provocação com HDM também foi reduzido significativamente pela administração de 15d-PGJ2. Em conjunto, nossos dados indicam que o tratamento com 15d-PGJ2 inibe alterações cruciais associadas à patogênese da asma, em modelos experimentais distintos da doença, demonstrando possuir grande potencial para controlar e reverter inflamação, hiperreatividade e remodelamento pulmonar desencadeados por provocação alérgica. / Asthma is a chronic pulmonary disorder characterized by inflammation, obstruction and airway remodeling, leading to symptoms such as wheezing, coughing and breathlessness. Asthma therapy is based on inhaled corticosteroids and short or long term-&#946;2 agonists. The treatment is limited by side effects and some refractory patients, justifying the study for new therapies. Studies have demonstrated that 15-deoxy-delta-12 ,14-prostaglandin J2 (15d-PGJ2), an endogenous ligand of peroxisome proliferator-activated receptor gamma (PPAR-&#947;) can reduce pro-inflammatory cytokines expression, providing a protective effect in diseases with this profile. The aim of this study was to evaluate the anti-inflammatory and antiasthmatic properties of 15d-PGJ2 in murine models of experimental asthma. A/J mice rats were sensitized on days 0 and 7 by subcutaneous (s.c.) injection , containing ovalbumin (OVA) and Al(OH)3, and challenged with 4 intranasal OVA instillations at weekly intervals. 15d-PGJ2 treatment (30 and 100 &#956;g/Kg) was performed 30 min before the challenges from the third antigen challenge. In another model, A/J mice were intranasally (i.n.) challenged with mite extract 3 times per week for 3 weeks. The administration of 15d-PGJ2 (30, 70 and 100 &#956;g /Kg, s.c. and 0.65, 1.5 and 2.3 &#956;g / animal, i.n.) were performed from the 3rd week, 30 min before the challenges. The analyzes were 24 h after the last challenge. Our results showed that in previously OVA-sensitized and challenged mice, administration of 15d-PGJ2 limited significantly (p <0.05), eosinophilic and neutrophilic inflammation and mucus production by goblet cells and sub-epithelial fibrosis, as well as airways hyperreactivity and IL-5 production. The reduction of bronchial epithelium and IL-13 and TNF-&#945; were observed only at the highest dose administered. In HDM model, inflammatory and remodeling parameters were attenuated in all administered doses of compound, whereas bronchial hyperresponsiveness was inhibited only at doses of 70 and 100 &#956;g/kg (s.c.) and 1.5 &#956;g/animal (i.n.). Serum cytokine levels were attenuated by subcutaneous treatment, but only IL-17, Eotaxin-1 and TNF-&#945; was inhibited by intranasal dose of 0.65 &#956;g/ animal. The increased expression of NF-kB induced by HDM challenge was also significantly reduced by the administration of 15d-PGJ2. Together, our data indicate that treatment with 15d-PGJ2 inhibits critical changes associated with the pathogenesis of asthma in different experimental models of the disease, demonstrating great potential to control and reverse pulmonary inflammation, hyperresponsiveness and remodeling triggered by allergen challenge.

O conhecimento da dinâmica da resistência de pragas a pesticidas é de fundamental importância para a implementação de estratégias de manejo da resistência. Desta forma, o principal objetivo do presente trabalho foi estudar a dinâmica da resistência de Brevipalpus phoenicis (Geijskes, 1939) ao acaricida dicofol em pomares comerciais de citros durante um período de aproximadamente três anos. Para tanto, foram realizados estudos para avaliar a resposta à pressão de seleção com dicofol em populações de B. phoenicis com diferentes freqüências iniciais de resistência e acompanhamento da freqüência de resistência na ausência de pressão seletiva. Alguns fatores que podem estar envolvidos na dinâmica da resistência de B. phoenicis a acaricidas como a capacidade de dispersão e a efetividade de hospedeiros alternativos como refúgio de indivíduos suscetíveis foram avaliados. Aplicações de dicofol em alguns talhões de citros resultaram em aumentos significativos na freqüência de resistência. A velocidade de restabelecimento da suscetibilidade na ausência de pressão seletiva variou entre os talhões. Em alguns talhões, o restabelecimento da suscetibilidade foi observado em um período de aproximadamente um ano. Em outros talhões, o restabelecimento da suscetibilidade não foi observado mesmo após 29 meses sem o uso do dicofol. Trabalhos de dispersão de B. phoenicis conduzidos em casa de vegetação mostraram que a capacidade de dispersão por caminhamento é limitada. Com a liberação de 6.000 ácaros em um determinado ponto, apenas 3% atingiram distâncias de 40 a 50 cm. As distâncias percorridas por este ácaro foram geralmente inferiores a 1 cm.dia-1. Em condições de laboratório, verificou-se que vento de 23 km.h-1 não foi capaz de arrastar ácaros da superfície de frutos; e velocidades de 30 e 40 km.h-1 foram capazes de arrastar menos de 1% da população de ácaros provenientes tanto de colônias novas quanto de colônias velhas. Trabalhos de dispersão em condições de campo mediante a utilização de armadilhas adesivas também comprovaram que a dispersão de B. phoenicis é bastante limitada quando comparada a de outras espécies de ácaros que ocorrem nos pomares de citros. De um total 2.420 e 661 ácaros coletados em dois talhões de citros na região de Descalvado-SP, apenas 0,45 e 11,80% dos ácaros pertenciam à família Tenuipalpidae, respectivamente. Para verificar a efetividade de outras plantas hospedeiras de B. phoenicis como refúgio de indivíduos suscetíveis, a variabilidade genética em populações de B. phoenicis provenientes de cercas-vivas de sansão-de-campo (Mimosa caesalpiniaefolia Benth.) localizadas próximas aos pomares de citros foi avaliada mediante estimativa da freqüência de resistência ao dicofol e análise molecular. Em geral, a freqüência de resistência em populações de B. phoenicis provenientes de sansão-de-campo foi inferior a de citros em um mesmo pomar. No entanto, a análise da variabilidade genética através da técnica de RAPD - PCR mostrou que as populações de B. phoenicis provenientes de sansão-do-campo são geneticamente diferentes das populações provenientes de citros. Portanto, baseado nos resultados obtidos conclui-se que o restabelecimento da suscetibilidade de B. phoenicis ao dicofol é bastante variável e a imigração de indivíduos suscetíveis de áreas não tratadas ou de hospedeiros alternativos é bastante limitada. / The knowledge of dynamics of pest resistance to pesticides is crucial for the implementation of resistance management strategies. Therefore, the main objective of this research was to study the dynamics of the resistance of Brevipalpus phoenicis (Geijskes, 1939) to the acaricide dicofol in commercial citrus groves during a 3-year period. Studies to evaluate the selection response to dicofol were conducted in populations of B. phoenicis with different initial resistance frequencies. The frequency of resistance was also estimated in the absence of selection pressure. Some factors that influence the resistance dynamics of B. phoenicis such as the dispersal capacity and the effectiveness of alternative hosts as refuges for susceptible individuals were also evaluated in this study. There was a significant increase in the frequency of dicofol resistance with applications of dicofol in some citrus field plots. The rate of reset to susceptibility in the absence of selection pressure varied from field to field. In some fields, the reset to susceptibility was observed within approximately 12-month period. On the other hand, the reset to susceptibility was not observed even after 29 months without using dicofol. Dispersal studies of B. phoenicis on sandy surface conducted under greenhouse conditions showed that dispersal capacity by crawling is limited. After the release of 6,000 mites in one point, only 3% reached distances of 40 to 50 cm from the release point. This mite moved less than 1 cm.day-1. Wind speed of 23 km.h-1 was not enough to carry mites from citrus fruit surface under laboratory conditions; and wind speeds of 30 and 40 km.h-1 were capable to carry less than 1% of mites either from new or old mite colonies on fruits. Mite dispersal studies with the use of sticky traps under field conditions also showed that B. phoenicis dispersal is very limited when compared to other mite species that occur in citrus groves. From a total of 2,420 and 661 mites collected from two citrus fields in Descalvado-SP region, only 0.45 and 11.80% of mites were in the Tenuipalpidae family, respectively. In order to evaluate the effectiveness of other host plants of B. phoenicis as refuges for susceptible individuals, the genetic variability in populations of B. phoenicis from "sansão-de-campo" (Mimosa caesalpiniaefolia Benth.) located near the citrus groves was evaluated by the estimation of resistance frequencies to dicofol and by molecular techniques. In general, the frequency of resistance to dicofol in populations of B. phoenicis from "sansão-de-campo" was lower than that from citrus groves. However, the analysis of genetic variability with RAPD-PCR revealed that B. phoenicis populations from "sansão-de-campo" were genetically different from citrus groves. Based on our results, we concluded that the reset of susceptibility of B. phoenicis to dicofol is very variable and the immigration of susceptible individuals from untreated areas or alternative hosts is very limited.

Les effets anti-allergiques des bactéries lactiques sont suggérés par plusieurs études épidémiologiques, des essais cliniques et des modèles expérimentaux d’allergie. Cependant, les propriétés immunomodulatrices des bactéries lactiques sont sous-exploitées par les stratégies vaccinales développées pour combattre l’allergie et les mécanismes empruntés par ces bactéries pour moduler l’allergie restent peu caractérisés. Dès lors, nous avons caractérisé les propriétés immunomodulatrices qu’exerce Lactobacillus plantarum NCIMB8826, une bactérie lactique modèle, sur la cellule dendritique étant donné le rôle déterminant de cette cellule sur la réponse allergique. Nous montrons que L. plantarum induit une forte sécrétion d’IL-12 p40, d’IL-12 p70, de TNF-a mais une faible production d’IL-10. Cette faculté à induire la sécrétion de cytokines polarisantes dépend de TLR2, de TLR9, de MyD88, de NF-kB, des MAPKs (en particulier JNK, p38 et ERK 1/2), de la composition de l’acide lipotéichoïque de L. plantarum et de CD14. Nous montrons aussi que l’ADN génomique de L. plantarum est un agoniste de TLR9 et que CD14 et CD36 facilitent la liaison de la cellule dendritique avec L. plantarum. Ensuite, nous avons évalué le potentiel vaccinal d’une coadministration L. plantarum + Der p 1 dans un modèle murin d’allergie à Der p 1. Cette formulation vaccinale prévient la production d’IgE Der p 1-spécifique et atténue l’éosinophilie pulmonaire tout en stimulant une forte production d’anticorps IgG2a Der p 1-spécifiques et d’IFN-g par les cellules spléniques. Ces effets bénéfiques nous ont conduit à élaborer une bactérie lactique recombinante dérivée de L. plantarum produisant Der p 1 pour la vaccination contre l’allergie aux acariens. La forme antigénique que nous avons réussi à faire produire par L. plantarum correspond à une protéine de fusion entre la Maltose Binding Protein de E. coli et ProDer p 1 (le zymogène de Der p 1), la présence de ce partenaire de fusion étant indispensable à la production de ProDer p 1. En prophylaxie, la vaccination par cette bactérie recombinante prévient la production d’anticorps IgE-Der p 1-spécifiques et stimule la production d’anticorps IgG2a spécifiques, reproduisant les effets de la coadministration L. plantarum + Der p 1. Elle réduit de manière drastique la production d’IL-5 des cellules spléniques et des cellules ganglionnaires médiastinales et prévient l’éosinophilie pulmonaire mais n’a pas d’effet sur l’hyperréactivité bronchique. Der p 1 étant un des allergènes d’acarien les plus immunodominants, cet ensemble de données montre donc que cette bactérie recombinante constitue un vaccin prophylactique prometteur pour la prévention de l’allergie aux acariens. Des résultats préliminaires obtenus à partir de cellules dendritiques humaines et lymphocytes T autologues montrent la forte capacité de cette bactérie recombinante à induire le développement d’une réponse Th1 fortement polarisée (production d’IFN-g en l’absence de production d’IL-4 et d’IL-5), ce qui suggère que l’utilisation de cette bactérie recombinante pourrait être envisagée pour le traitement de l’allergie chez l’homme.

The major allergen Der p 1 of the house dust mite Dermatophagoides pteronyssinus is a papain-like cysteine protease (CA1) associated to the development of allergic diseases such as asthma, rhinitis or atopic dermatitis. This allergen is expressed as an inactive precursor, called proDer p 1, formed by a 25 kDa catalytic domain downstream to an 10 kDa N-terminal propeptide, which blocks the active site cleft. The propeptide of Der p 1 exhibits a specific fold, which makes it unique in the CA1 propeptide family as it is characterised by the presence of four alpha helices and the absence of ERFNIN motif. In this study, we investigated the activation steps involved in the maturation of recombinant proDer p 1 expressed in Pichia pastoris under acidic conditions and we studied the influence of acidic pH on the structure of both propeptide and catalytic domain. Therefore, we characterized the interaction between the propeptide and mature Der p 1 at different pH values in terms of activity inhibition, structural stability and proteolytic susceptibility. According to our results, the auto-activation of proDer p 1 is a multistep mechanism, characterized by at least two intermediates (ATFE- and SNGG-) corresponding to the loss of the first and second propeptide alpha helices, respectively. The propeptide strongly inhibits unglycosylated and glycosylated recombinant Der p 1 (KD= 7 nM) at neutral pH. This inhibition is pH dependent, decreasing from pH 7 to pH 4 and can be related to structural changes of the propeptide initiated by the protonation of the aspartate residue of Lys17-Asp51-Tyr19 structural triad presents within the propeptide N-terminal domain. This protonation triggers conformational changes of the first propeptide alpha helix leading to an increase of the propeptide flexibility, an increase of its proteolytic sensitivity and the formation of a molten globule state. In addition, we compare mature protease, zymogen and propeptide pH unfolding and stability and highlights that the presence of the propeptide does not influence the catalytic domain pH unfolding and stability as the propeptide displays a weaker pH stability than the protease domain. These results confirmed that the propeptide unfolding is the key event of the activation process. Finally, we unravel the intermolecular contribution of mature Der p 1 in the activation process and highlights that activation of the precursor can be achieved, under acidic conditions, by intermolecular process but initial auto-activation most probably occurs through an intramolecular process or by the proteolysis by the catalytic domain of another zymogen in which the propeptide is unfolded. According to our results, we proposed that activation of the zymogen at pH 4 reflects a compromise between activity preservation and propeptide unfolding and that the location of the activation sites on the propeptide structure is a compromise between sequence recognition specificity and proteolytic susceptibility of the corresponding area.