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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Comparison of Hypersensitivity Reaction Incidence to Carboplatin in Patients with Ovarian, Fallopian Tube, or Primary Peritoneal Cancer with or without the BRCA1 or BRCA2 Mutations

Garcia, Andrew, Corey Frahm January 2017 (has links)
Class of 2017 Abstract / Objectives: The specific aims of this project were to evaluate the incidence of carboplatin HSR in patients with the BRCA1 or BRCA2 mutations compared to those without these mutations. Secondary objectives were to identify carboplatin cycles where reactions occurred, grade of reaction, and treatment outcomes. Methods: This retrospective chart review included 167 ovarian, fallopian tube, and primary peritoneal cancer patients at the University of Arizona Cancer Center who underwent a regimen with carboplatin from 2013-2015. Results: 126 out of 167 patients were analyzed. HSR occurred in 4 patients with BRCA mutations, and in 9 patients without mutations, though incidence was not significant with respect to the groups (3.1% versus 17.4%, P=0.5291). Overall, there were 11 grade 1 reactions, 14 grade 2 reactions, and 16 grade 3 reactions to carboplatin. Conclusions: Presence of a BRCA1/2 mutation was not associated with a higher incidence of HSR in carboplatin. More studies are needed to clarify the impact of BRCA mutations on developing carboplatin HSR.
2

Low Rate of Cetuximab Hypersensitivity Reactions in Northeast Tennessee: An Appalachian Effect?

Adams, Brooke C., Street, Sierra D., Crass, Melanie, Bossaer, John B. 20 November 2015 (has links)
Purpose: Cetuximab is a monoclonal antibody with a known risk of hypersensitivity reactions. Early studies showed hypersensitivity reaction rates of 3%, but there appears to be a higher incidence in the southeastern United States. To confirm the findings from nearby institutions that cetuximab-associated hypersensitivity reactions occur in approximately 20% of patients in the southeastern United States. Methods: A retrospective chart review was conducted at Johnson City Medical Center in Johnson City, Tennessee. Each patient’s first infusion was analyzed for hypersensitivity reaction, as well as for demographic information such as allergy and smoking history, pre-medications, and malignancy type. Results: Data from the first infusion of cetuximab were collected for a total of 71 patients with various malignancies. The overall rate of grade 3 or higher hypersensitivity reaction was 1.4%, and total rate of hypersensitivity reaction was 8.5%. These findings more closely correlate to the early clinical trials and package insert. Both severe (p = 0.001) and any-grade (p = 0.002) hypersensitivity reaction occurred less frequently in one Southeastern Appalachian medical center compared to academic medical centers directly to the east and west. Conclusions: Patients in southern Appalachia may be less likely to develop cetuximab hypersensitivity reactions compared to surrounding areas in the Southeastern U.S. These results lend support to the theory that exposure to lonestar ticks (Amblyomma americanum) may be responsible for the development of IgE antibodies to cetuximab that cause hypersensitivity reactions. The development of quick and reliable bedside predictors of cetuximab hypersensitivity reactions may aid clinicians considering the use of cetuximab.
3

Caracterização de um isolado de Bean rugose mosaic virus e busca por fontes de resistência em Phaseolus vulgaris / Characterization of an isolate of Bean rugose mosaic virus and search for sources of resistance in Phaseolus vulgaris

Cândida, Daniella Vieira 31 March 2017 (has links)
Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2017-05-10T12:53:52Z No. of bitstreams: 2 Tese - Daniella Vieira Cândida - 2017.pdf: 1821838 bytes, checksum: a4d863f3de465899a5c2160c17565020 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-05-10T12:54:49Z (GMT) No. of bitstreams: 2 Tese - Daniella Vieira Cândida - 2017.pdf: 1821838 bytes, checksum: a4d863f3de465899a5c2160c17565020 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-05-10T12:54:49Z (GMT). No. of bitstreams: 2 Tese - Daniella Vieira Cândida - 2017.pdf: 1821838 bytes, checksum: a4d863f3de465899a5c2160c17565020 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-03-31 / Abstract: In 2013, common bean plants of the cultivar Pérola were found in an experimental field belonging to Embrapa Arroz e Feijão Lat. 16 ° 28 '00 "(S); Long. 49 ° 17 '00 "(W); (GO) presenting leaf distortion, mosaic and blistering. The sample analysis by electron microscopy detected the presence of typical Comovirus genus viral particles, thus, the identification of the virus species through sequencing became essential and the search for control alternatives, due to the damage potential of Bean rugose mosaic virus (BRMV) to bean production fields. Therefore, the present work had as objectives: (1) the molecular characterization of BRMV-GO, an isolate from common bean and (2) the search for bean accessions resistant to this viral species. For germplasm selection, 172 accessions were analyzed by means of mechanical inoculation and visualization of symptoms at 5, 21 and 30 days after inoculation. The range of hosts was analyzed by inoculation of 15 typical indicator species. Soya plants (Cv. Savana, Cv. Cristalina, Cv. Doko and Cv. Conquista) and pea (Cv. Mikado and Cv. Triofin) were also analyzed for reactions to BRMV-GO. Leaves of the infected plants were used for detection of BRMV-GO through RT-PCR. Of the 172 analyzed accessions, 168 behaved as susceptible, 3 accessions: BGF0011750 (cv. Mulatinho), BGF0000880 (cv. Rico 23) and BGF0001083 (cv Rico 23) reacted to BRMV-GO with vein and petioles necrosis followed by death; 1 access, BGF0003174 cv. IPA5047, showed a hypersensitivity reaction. The two species of the indicator Chenopodium amaranticolor and C. quinoa reacted with local necrotic lesions and no systemic infection confirmed by RT-PCR. Soybean plants reacted as susceptible and all pea plants showed tip burning. For molecular characterization, complete genome sequencing was performed by Sanger method using the primer walking strategy, the 5 'and 3' ends were obtained by RACE method. Genome sizes were 5906 nucleotides with a 1856 amino acid polyprotein for RNA 1 and 3688 nucleotides with a polypeptide of 1096 amino acids for RNA 2. The nucleotide identity between BRMV-GO and BRMV-Paraná isolates was 92.7% for RNA 1 and 90.5% for RNA 2. The highest percentage of identity obtained by amino acids sequence alignment of the polymerase (RdRp) and the capsid protein (CP) was 63% (RdRp) and 66% (CPs) with Bean pod mottle virus (BPMV), however, the ICTV delimits 80% (RdRp) and 75% (CP) identity to be part of the Comovirus genus, however, these results agree with the results obtained in another work with a Paraná isolate, corroborating that BRMV is a distinct species among this genus. Phylogenetic analysis of regions RdRp and CPs showed that BRMV-GO and BRMV-Paraná do not have significant differences and revealed higher indexes of identity with BPMV, both for RNA 1 and RNA 2. The complete sequences of RNA 1 and RNA 2 were deposited on Genbank under accession numbers KY622124, KY622125, respectively. / Resumo: Em 2013, plantas de feijão-comum da cultivar Pérola foram observadas em um campo experimental da Embrapa Arroz e Feijão Lat. 16° 28’ 00”(S); Long. 49° 17’ 00”(W); (GO) apresentando deformação foliar, mosaico em desenho e bolhosidade. A análise das amostras por microscopia eletrônica detectou a presença de partículas virais típicas do gênero Comovirus, sendo assim necessária a identificação da espécie do vírus através de sequenciamento e a busca por alternativas para o controle, devido ao potencial de dano da espécie Bean rugose mosaic virus (BRMV) para a cultura do feijão. Por isso, o presente trabalho teve como objetivos: (1) a caracterização molecular do isolado BRMV-GO, proveniente de feijoeiro e (2) a busca por acessos de feijoeiro resistentes à essa espécie viral. Para a seleção de germoplasma, 172 acessos foram analisados por meio de inoculação mecânica e visualização dos sintomas aos 5, 21 e 30 dias após a inoculação. A gama de hospedeiras foi analisada mediante inoculação de 15 espécies indicadoras típicas. Plantas de soja (cv. Savana, cv. Cristalina, cv. Doko e cv. Conquista) e ervilha (cv. Mikado e cv. Triofin) também foram analisadas quanto à reação ao BRMV-GO. Folhas das plantas infectadas foram usadas para detecção de BRMV-GO via RT-PCR. Dos 172 acessos analisados, 168 se comportaram como suscetíveis, 3 acessos: BGF0011750 (cv. Mulatinho), BGF0000880 (cv. Rico 23) e BGF0001083 (cv. Rico 23) reagiram ao BRMV-GO com necroses nas nervuras e pecíolos seguida de morte; 1 acesso, BGF0003174 cv. IPA5047, apresentou reação de hipersensibilidade. As duas espécies de indicadoras Chenopodium amaranticolor e C. quinoa reagiram com lesões locais necróticas não ocorrendo infecção sistêmica confirmada por meio de RT-PCR. As plantas de soja foram suscetíveis e todas as plantas de ervilha apresentaram queima do topo. Para a caracterização molecular, o sequenciamento completo do genoma foi realizado pelo método de Sanger usando a estratégia primer walking, as extremidades 5’ e 3’ foram obtidas pelo método RACE. Os tamanhos dos genomas foram de 5906 nucleotídeos com uma poliproteína de 1856 aminoácidos para RNA 1 e para RNA 2 foi de 3688 nucleotídeos com uma poliproteína de 1096 aminoácidos. A identidade de nucleotídeos entre os isolados BRMV-GO e BRMV-Paraná foi de 92,7 % para RNA 1 e 90,5% para RNA 2. A maior porcentagem de identidade obtida através do alinhamento de sequências de aminoácidos da polimerase (RdRp) e da proteína do capsídeo (CP) foi de 63% (RdRp) e 66% (CPs) com Bean pod mottle virus (BPMV), entretanto, o ICTV delimita para membros do gênero Comovirus uma identidade de 75% para aminoácidos (CPs) e 80% para a RdRp, no entanto, esses resultados estão de acordo com os resultados obtidos em outro trabalho com um isolado do Paraná, corroborando que BRMV é uma espécie distinta dentro do gênero. As análises filogenéticas das regiões RdRp e CPs mostraram que BRMV-GO e BRMV-Paraná não possuem diferenças significativas e revelou maiores índices de identidade com BPMV, tanto para o RNA 1 como para o RNA 2. As sequências completas do RNA 1 e RNA 2 foram depositadas no Genbank com os números de acesso KY622124, KY622125, respectivamente.
4

Antidote or Poison: A Case of Anaphylactic Shock After Intra-Articular Corticosteroid Injection

Sethi, Pooja, Treece, Jennifer, Onweni, Chidinma, Pai, Vandana 29 August 2017 (has links)
Although glucocorticoids are often used as an adjunct to epinephrine to treat anaphylactic shock, glucocorticoids can also be a rare cause of anaphylactic shock. Only through the administration of a challenge dose of different glucocorticoids and different substrates that glucocorticoids are delivered in can the determination be made about which glucocorticoid or accompanying solvent may be the culprit which caused the anaphylactic reaction. These challenge tests should only be performed in a controlled environment as repeat anaphylaxis is a risk, especially if the patient has a history of glucocorticoid-induced anaphylaxis.
5

Histologia, bioquímica e herança da resistência do genótipo de algodoeiro TX25 a Meloidogyne incognita, raça 3 / Histology, biochemistry and resistance inheritance of cotton genotype TX25 to Meloidogyne incognita, raça 3

Alves, Gleina Costa Silva 31 January 2014 (has links)
Submitted by Erika Demachki (erikademachki@gmail.com) on 2015-10-20T19:35:32Z No. of bitstreams: 2 Tese - Gleina Costa Silva Alves - 2014.pdf: 1815894 bytes, checksum: a6e6bf5cc3cb4021e3377efd18c007d1 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Erika Demachki (erikademachki@gmail.com) on 2015-10-20T19:36:55Z (GMT) No. of bitstreams: 2 Tese - Gleina Costa Silva Alves - 2014.pdf: 1815894 bytes, checksum: a6e6bf5cc3cb4021e3377efd18c007d1 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2015-10-20T19:36:55Z (GMT). No. of bitstreams: 2 Tese - Gleina Costa Silva Alves - 2014.pdf: 1815894 bytes, checksum: a6e6bf5cc3cb4021e3377efd18c007d1 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-01-31 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Meloidogyne incognita race 3 is a limiting factor for cotton crop, and the genetic resistance is one of the most desirable control measures for being environmentally and economically suitable. The objective of this research was to study the resistance inheritance of cotton genotype TX 25 to M. incognita race 3 and identify the resistance mechanism. Two experiments for phenotyping were conducted under greenhouse conditions, using two parental genotypes (susceptible FM 966 and resistant TX 25), backcross, generation F1 and generation F2. The plants were maintained for 120 days after inoculation (DAI). Plants were evaluated for gall index, egg mass index and reproduction factor. Histopathological study was conducted and the root systems evaluated at 2, 4, 7, 9, 11, 16, 18, 21 , 23, 29, 34 and 45 DAI. The roots werestained with acid fuscin to assess the nematode penetration. Rootlet cuts were made to observe the changes caused by the presence of the nematode in the root system of susceptible and resistant cotton cultivar. Biochemical tests were also conducted with the two parental genotypes. In this essay, the plants were kept in the greenhouse and inoculated with eggs and J2 of M. incognita. Assessments occurred at eight, 24 and 35 days after inoculation. The genotypes were compared biochemically as to total phenols, flavonoids, liquid chromatography and magnetic resonance imaging. The genotype TX 25 showed resistance to M. incognita race 3 when attacked by the nematode with the plants producing a hypersensitivity reaction. The crosses derived from TX 25 and FM 966 showed olygogenic resistance. TX 25 also shows glycosylated flavonoids and sugars that provide resistance to Meloidogyne incognita race 3. / Meloidogyne incognita, raça 3 é um dos fatores limitantes da cultura do algodoeiro, e a resistência genética é uma medida de controle das mais desejáveis por ser ambientalmente preservadora e não resultar em gastos adicionais ao produtor. O objetivo da presente pesquisa foi estudar a herança da resistência do genótipo de algodão TX 25 a M. incognita, raça 3, e identificar o mecanismo de ação. Foram conduzidos dois experimentos de fenotipagem em casa de vegetação, com dois parentais, um suscetível FM 966 e um resistente TX 25, retrocruzamento, geração F1 e geração F2, onde as plantas foram mantidas por 120 dias após serem inoculadas com o nematóide. Posteriormente avaliou-se índice de galhas, índice de massa de ovos e fator de reprodução. Experimento histopatológico foi conduzido onde as raízes de algodoeiro TX 25 e FM 966 foram avaliadas aos 2, 4, 7, 9, 11, 16, 18, 21, 23 e 29, 34 e 45 dias após a inoculação, com coloração do sistema radicular com fuscina ácida para avaliar a penetração. Foram feitos cortes nas radicelas para observar as alterações causadas pela presença do nematóide no sistema radicular da cultivar suscetível e genótipo resistente. Ensaios bioquímicos com os dois parentais também foram realizados. Nesse ensaio as plantas foram mantidas em casa de vegetação e inoculadas com ovos e J2 de M. incognita. As avaliações ocorreram aos oito, 24 e 35 dias após a inoculação. Nessas avaliações os parentais foram comparados bioquimicante quanto aos fenóis totais, flavonóides, cromatografia líquida e ressonância magnética. O genótipo TX 25 apresenta resistência a M. incognita raça 3, quando incitado por esse nematóide a planta produz reação de hipersensibilidade. A geração F2 oriundas de TX 25 e FiberMax 966 apresentou uma herança de caráter provavelmente oligogênica. O genótipo TX 25 apresenta flavonóides glicolisados e açúcares, que conferem resistência a Meloidogyne incognita, raça 3. O TX 25 é um material indicado como fonte de resistência para os programas de melhoramento genético.
6

Estudo da ação imunobiológica do laser de baixa intensidade sobre modelo de hipersensibilidade tardia à ovalbumina.

Oliveira, Rodrigo Guerra de 30 June 2010 (has links)
Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-10-04T18:59:22Z No. of bitstreams: 1 rodrigoguerradeoliveira.pdf: 1453787 bytes, checksum: 629265209ed579deb47cf4c16a39a88a (MD5) / Approved for entry into archive by Diamantino Mayra (mayra.diamantino@ufjf.edu.br) on 2016-10-05T10:52:32Z (GMT) No. of bitstreams: 1 rodrigoguerradeoliveira.pdf: 1453787 bytes, checksum: 629265209ed579deb47cf4c16a39a88a (MD5) / Made available in DSpace on 2016-10-05T10:52:32Z (GMT). No. of bitstreams: 1 rodrigoguerradeoliveira.pdf: 1453787 bytes, checksum: 629265209ed579deb47cf4c16a39a88a (MD5) Previous issue date: 2010-06-30 / O objetivo deste estudo foi avaliar o efeito do Laser de Baixa Intensidade (LLLT) em um modelo de Hipersensibilidade Tardia (RHT). LLLT vem sendo estudado há algum tempo e seus efeitos clínicos, aplicados no tratamento de várias doenças. O laser tem sido testado em diferentes modelos experimentais, mas seus efeitos permanecem obscuros. Tentou-se avaliar os efeitos do LLLT na RHT à ovalbumina (OVA), uma proteína que vem sendo utilizada como antígeno para sensibilizar cobaias. Esse é um modelo amplamente utilizado para avaliar os efeitos de substâncias com potencial para modular o sistema imunológico e as reações inflamatórias. Camundongos Balb/C foram divididos, randomicamente, em quatro grupos, (I) imunizado, não tratado e desafiado (n=6), (II) não imunizado, não tratado e desafiado (n=6); (III) imunizado, tratado com Azatioprina e desafiado (n=6); e o grupo (IV), imunizado, tratado com LLLT e desafiado (n=6). Passadas 48 horas do desafio, os animais foram submetidos a avaliação do edema da pata e foram eutanasiados para análise do coxim plantar. Testes de proliferação foram realizados (espontâneos, na presença de cocanavalina A e ovalbumina) para determinar a produção, em cultura de células, de TNF- , INF- and IL-10. Análises imunohistoquímicas para expressão de COX-2 também foram realizadas. No grupo de animais irradiados com laser e naqueles tratados com AZA, a medida da espessura da pata foi significativamente menor em comparação à do grupo controle. Tal fato foi acompanhado da redução significativa na densidade do infiltrado inflamatório, assim como a redução significativa nos níveis de TNF- , INF- and IL-10 e na expressão de COX-2. Nossos resultados sugerem que o tratamento com laser possui efeito imunomodulador na RHT à OVA e podem contribuir para a imunoterapêutica do transplante renal. / The aim of this study was to evaluate the effect of Low Level Laser Therapy (LLLT) in an experimental model of delayed hypersensitivity reaction (DTH). LLLT has been studied for some time and its clinical effects have been used to treat numerous diseases. LLLT has been tested in different experimental models and some of its effects have yet to be explained. We tried to assess the effects of LLLT on the DTH reaction to ovalbumin (OVA), a protein that has been used as an antigen to sensitize lab animals. This is a broadly used experimental model to assess the effects of substances that have the potential to modulate the immune system and inflammatory reactions. Balb/C mice were randomly divided into four groups, (I) immunized, untreated and challenged (n=6), (II) not immunized, untreated and challenged (n=6); (III) immunized, treated with Azathioprine (AZA) and challenged (n=6); and group (IV) immunized, treated with LLLT and challenged (n=6). Forty-eight hours after the challenge, the animals were submitted to a paw edema check and were euthanized for histopathology analysis of their plantar pads. Proliferations tests were made (spontaneous, in the presence of concanavalin A and Ovalbumin) to determine the production in cell cultures of TNF- , INF- and IL-10. Immunohistochemical analyzes for expression of COX-2 were also performed. In the group of animals irradiated with lasers and those treated with AZA, footpad thickness measurements were significantly reduced in comparison to the control group. This was accompanied by a very significant drop in the density of the inflammatory infiltration as well as by a significant reduction in the levels of concentration of TNF- , INF- and IL-10 and in the expression of COX-2. Our results suggest that treatment with LLLT has immunomodulatory effect and may have important contribution to the immunotherapeutic of kidney transplants.
7

Réactions d'hypersensibilité immédiate en anesthésie : épidémiologie et risques environnementaux / Immediate hypersensitivity reactions in anesthesia : Epidemiology and environmental risks

Dong, Siwei 14 December 2012 (has links)
Les réactions d'hypersensibilité immédiate survenant au cours de l'anesthésie demeurent un sujet de préoccupation majeure pour les anesthésistes. Le médicament le plus fréquemment responsable d'une anaphylaxie sont les curares. Le but de ce travail est de préciser l'épidémiologie actuelle et d'identifier certains facteurs environnementaux susceptibles d'expliquer la fréquence élevée des réactions allergiques aux curares qui peuvent survenir en l'absence de toute exposition préalable. Dans la première partie de la thèse, une enquête nationale décrivant l'épidémiologie des réactions anaphylactiques peranesthésiques entre 2005 et 2007 en France. Elle a confirmé l'intérêt du dosage d'IgE spécifique anti-curare. La survenue de réactions d'hypersensibilité immédiate allergique à l'induction anesthésique lors d'une première exposition aux curares a conduit à évoquer l'existence d'une sensibilisation croisée avec des substances présentes dans l'environnement et possédant un ammonium quaternaire. Différentes hypothèses ont été proposées, notamment une exposition aux produits cosmétiques et/ou désinfectants, une exposition à des antigènes de type phosphorylcholine rencontrés dans les levures, ou certains parasites. Dans la deuxième partie de la thèse, deux populations professionnelles, des apprentis coiffeurs exposés à des agents chimiques contenant des ions ammoniums, et des boulanger-pâtissiers exposés à des allergènes de levures et de parasites, ont été comparées avec la population générale à la recherche de différences concernant la prévalence des anticorps IgE spécifique anti-curare. Au terme de ce travail, l'exposition à des produits cosmétiques chez les coiffeurs apparaît comme un facteur de risque de développer des anticorps IgE spécifique anti-curare, démontrant pour la première fois l'hypothèse du lien entre l'exposition répétée aux produits cosmétiques contenant l'ammonium quaternaire et la sensibilisation croisée aux curares / Immediate hypersensitivity reactions occurring during anesthesia remains a major concern for anesthesiologists. The drugs most frequently responsible for anaphylaxis are neuromuscular blocking agents (NMBAs). The purpose of this study is to determine the current epidemiology and to identify environmental factors that may explain the high frequency of allergic reactions to neuromuscular blocking agents that can occur without any prior exposure. In the first part of the thesis, a national survey describing the epidemiology of anaphylaxis during anesthesia between 2005 and 2007 in France was carried out. The value of testing specific IgE against NMBA was confirmed.The occurrence of immediate allergic hypersensitivity reaction when first exposure to an NMBA during induction led to evoke the existence of cross-sensitization with substances present in environment and having a quaternary ammonium compound. Different hypotheses have been proposed including exposure to cosmetics and / or disinfectants, exposure to antigens encountered in phosphorylcholine yeasts, or parasites, or exposure to a derivative opiate, pholcodine. In the second part of the thesis, two occupational populations, hairdressers apprentice exposed to chemical agents containing ammonium ions, baker and pastry maker exposed to allergens yeast and parasites were compared with the general population to search for differences regarding the prevalence of specific IgE antibody against NMBA. At the end of this work, hairdressers exposed to cosmetics agents appears to be a risk factor for developing specific IgE antibodies against NMBA and we demonstrated for the first time the hypothesis of the ross-sensitization between repeated exposures to cosmetics products containing quaternary ammonium and NMBAs
8

Avaliação comparativa das proteinas de fusão cmx e ecmx no teste de mantoux para o diagnóstico de tuberculose / Comparative evaluation of fusion proteins cmx and ecmx by mantoux technique for tuberculosis diagnosis

Sánchez, Tatiana Marlene Galvez 20 February 2017 (has links)
Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2017-03-24T11:22:47Z No. of bitstreams: 2 Dissertação - Tatiana Marlene Galvez Sánchez - 2017.pdf: 3124839 bytes, checksum: 942aa6d4fa9aa30babb4eb0d7e04a805 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-03-24T12:39:25Z (GMT) No. of bitstreams: 2 Dissertação - Tatiana Marlene Galvez Sánchez - 2017.pdf: 3124839 bytes, checksum: 942aa6d4fa9aa30babb4eb0d7e04a805 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-03-24T12:39:25Z (GMT). No. of bitstreams: 2 Dissertação - Tatiana Marlene Galvez Sánchez - 2017.pdf: 3124839 bytes, checksum: 942aa6d4fa9aa30babb4eb0d7e04a805 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-02-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Outro / Tuberculin skin test (TST) identifies a previous exposed to M. tuberculosis (Mtb) using an intradermal inoculation of purified protein derivates (PPD) that result in a delayed hypersensitivity reaction (DTH). Interferon-gamma release assay (IGRA) was suggested to replace TST. The IGRA uses antigens, ESAT-6 and CFP-10, absent in all BCG strains and some non tuberculous mycobacteria (NTM). However, reproducibility and high cost were limitations for endemic countries. For this reason, the development of new diagnose test for latent TB is necessary. Fusion proteins developed by our group has been recognized by the immune response generated by the infection with Mycobacterium tuberculosis. Thus the aim of this work was to evaluate the capacity of CMX or ECMX to be used in a Skin test for tuberculosis. BALB/c mice infected with Mtb were euthanized forty-five days after infection. Spleens, lungs and draining lymph nodes of infected mice were processed and evaluated by flow cytometry Both CD4 and CD8 IFN+ cells were able to recognize rCMX and rECMX. The skin test followed an evaluation of thickness/swelling ≥ PPD 2UT (positive control) to consider positive DTH. Based on thickness, at 24 h, rCMX 25μg (0.37±0.02) and rECMX 15-25μg (0.38±0.03/0,62±0,12) induced a positive DTH response. At 48h, rCMX 25μg (0.28±0.03) and rECMX 25μg (0.5±0.04) induced also a positive DTH reaction. In conclusion, fusion proteins rCMX and rECMX are recognized by infected mice with Mtb and skin test using rECMX 25μg induced better DTH response that of conventional PPD. / A prova tuberculínica (PT) é um teste cutâneo que identifica a exposição prévia ao M. tuberculosis (Mtb), mediante a inoculação via intradérmica do derivado protéico purificado (PPD) de Mtb, o que resulta em uma reação de hipersensibilidade do tipo tardia (DTH). O ensaio de liberação de IFN-γ (IGRA) foi indicado para substituir a PT. O IGRA usa os antígenos ausentes na BCG e algumas micobactérias não causadoras de TB (MNT), ESAT-6 e CFP-10. Porém, apresenta falta de reprodutibilidade e alto custo quando usado em populações endêmicas para TB. Diante disso, o desenvolvimento de novos testes de diagnóstico é necessário. Nosso grupo desenvolveu proteínas de fusão que são reconhecidas por linfócitos gerados pela infecção com Mtb. Assim, o trabalho propõe avaliar a utilização das proteínas rCMX e rECMX no desenvolvimento de um teste cutâneo de diagnóstico para tuberculose. Camundongos BALB/c foram infectados com Mtb H37Rv. Após 45 dias, a infecção induziu linfócitos T CD4+ e CD8+ produtores de IFN-γ específicos para rCMX e rECMX no baço, pulmões e linfonodos drenantes. Enquanto ao teste cutâneo realizado 45 dias após a infecção, a leitura de espessura/inchaço ≥ PPD 2UT (controle positivo) indicou uma reação de DTH positiva. Avaliando a espessura 24h após o inóculo, rCMX 25μg (0.37±0.02) e rECMX 15-25μg (0.38±0.03/0,62±0,12) induziram reação de DTH positiva. As 48h, rCMX 25μg (0.28±0.03) e rECMX 25μg (0.5±0.04) também apresentaram reação positiva. Enquanto o inchaço as 24h, só a rECMX apresentou DTH positiva. Em conclusão, este trabalho mostra que as proteínas rCMX e rECMX são reconhecidas pela resposta celular de camundongos infectados com Mtb, e quando usadas no teste cutâneo induziram reação de DTH positiva comparável e até superior ao PPD convencional. Dessa forma, é recomendada a avaliação das proteínas de fusão em outros modelos animais e posteriormente em humanos.

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