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Indiana University-Purdue University Indianapolis (IUPUI) / Inflammation of the urethra—urethritis—is commonly diagnosed in men and women who have sexually transmitted infections (STI). Characteristic signs and symptoms of urethritis include urethral discharge and burning pain during urination (dysuria). However, these findings are non-specific and can be elicited by STI for which optimal treatment approaches differ. We wanted to investigate if immunoglobulins (antibodies) in the urine of men with acute urethritis could determine the etiologies of these cases. Previously, we conducted an observational case-control study of biological males to compare the urethral microbiota of participants with unambiguous, laboratory-confirmed urethritis (cases) and participants without urethral inflammation (controls). This revealed that nearly 2 in 5 men with nongonococcal urethritis tested negative for all common STI. We identified atypical urethral pathogens in approximately 1/3 of these STI-negative individuals using shotgun metagenomic sequencing. However, we did not detect microorganisms suspected to be urethral pathogens in the remaining 2/3 of STI-negative participants. We hypothesized that these men with “pathogen-negative” urethritis had persisting inflammation from a recent STI that already cleared spontaneously by the time of testing. We observed that urine IgA antibodies against Chlamydia trachomatis (Ctr) infectious particles were significantly more prevalent among men with pathogen-negative urethritis compared to controls. In contrast, we found that the prevalence of urine anti-Ctr IgA was similar between controls and urethritis cases with atypical infections. However, our efforts to detect antibodies against another common STI, Mycoplasma genitalium (Mgen), were complicated by low abundance in urine and the unexpected prevalence of Mgen-reactive antibodies among controls. Collectively, our results suggest that signs and symptoms of urethritis can continue after the causative STI(s) have been eliminated. Furthermore, male urine represents a practical, non-invasive source of pathogen-reactive antibodies that could be evaluated using point-of-care diagnostic tests to elucidate urethritis etiologies. Importantly, our results also suggest that sexual partners of men with pathogen-negative, nongonococcal urethritis are an unrecognized chlamydia reservoir. / 2024-05-22

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Shakespeare references humoral medical theory and social definitions of gender throughout much of his work. His references to medical practices like purging, the siphoning of excessive emotional fluids to bring the body into balance, are more than allusions to medical theories. Shakespeare's works unveil and challenge early modern approaches to emotional experience, most particularly when it comes to traumatic experiences that overwhelm comprehension. In Titus Andronicus (1592), The Rape of Lucrece (1593), Hamlet (1603), King Lear (1608), and Macbeth (1606), Shakespeare invokes humoral theory to articulate the early modern traumatic experience and to criticize the efficacy of purging in representations of trauma. For Shakespeare, the siphoning of destabilized emotions, through metaphorical and rhetorical practices, has dangerous consequences for bodies coded as feminine.

Foram imunizados 27 macacos-prego (Cebus apella), por via intramuscular, com vacina anti-rábica inativada, produzida a partir de cérebros de camundongos lactentes (VARCCL), empregada nas campanhas de prevenção da raiva animal de cães e gatos. Os animais permaneceram em cativeiro, durante o período de junho de 1995 a junho de 1997, sendo divididos em 3 grupos experimentais e vacinados conforme o esquema: grupo I, com 3 doses a intervalo de 30 dias; o II, com 2 doses a intervalo 30 dias e reforço aos 210 dias; e, o III com uma dose e reforço aos 210 dias. A revacinação anual foi realizada em todos os animais aos 365 dias. As amostras de soros foram obtidas aos 0, 30, 60, 90, 150, 210, 240, 300, 365, 395, 545 e 730 dias, e armazenadas à temperatura de -20ºC, e a dosagem dos anticorpos realizada através do teste simplificado da inibição da fluorescência. Verificou-se que os animais do grupo I apresentaram soroconversão mais duradoura (12232 dias) do que os dos grupos II e III (p<0,05) após a vacinação inicial, e os animais do grupo I apresentaram soroconversão mais duradoura (183,6120,6 dias) do que os dos grupos II (p<0,05), após a revacinação anual aos 365 dias. A VARCCL induziu a resposta imune nos macacos-prego, após vacinação e revacinação, respectivamente, em 81,4 por cento e 76,0 por cento dos animais; com produção de anticorpos neutralizantes, iguais ou superiores a 0,5 UI/ml, porém, de curta duração; não constituindo assim, imunógeno apropriado para ser utilizado na rotina de imunização destes animais de difícil lide, mantidos em cativeiro / Twenty-seven capuchin monkeys (Cebus apella) were intramuscularly immunized with inactivated suckling mouse brain rabies vaccine (SMBV) employed in campaigns for animal rabies prevention in dogs and cats. The animals were kept confined from June, 1995 to June, 1997. They were divided into 3 experimental groups and vaccinated according to the following scheme: group I, with 3 doses within a 30-day interval; group II, with 2 doses within a 30-day interval and a booster dose at the 210th day; and, group III; with a single dose and a booster dose at the 210th day. All animals were given the annual re-vaccination at the 365th day. Sera samples were obtained at the 0th, 30 th, 60th, 90 th, 150 th, 210 th, 240 th, 300 th, 365 th, 395 th, 545 th e 730 th days and kept stored at 20ºC. The antibodies dosage was carried out through the simplified inhibition fluorescent test. The following results were observed: animals belonging to group I had longer humoral immune response (12232 days) than the ones belonging either to group II or group III (p<0.05) after initial vaccination; animals of group I presented longer humoral immune response (183.6120.6 days) than the ones of the group II (p<0.05) after the annual re-vaccination at the 365th day. The SMBV induced humoral immune response in capuchin monkeys after vaccination and re-vaccination in respectively 81.4 per cent and 76.0 per cent of the animals, producing neutralizing antibodies equal to or higher than 0.5 IU/ml; however, they were short-lasting, being therefore not appropriate as an immunogen to be used routinely in the immunization of these animals which are difficult both to be deal with and to be held confined

Foram imunizados 27 macacos-prego (Cebus apella), por via intramuscular, com vacina anti-rábica inativada, produzida a partir de cérebros de camundongos lactentes (VARCCL), empregada nas campanhas de prevenção da raiva animal de cães e gatos. Os animais permaneceram em cativeiro, durante o período de junho de 1995 a junho de 1997, sendo divididos em 3 grupos experimentais e vacinados conforme o esquema: grupo I, com 3 doses a intervalo de 30 dias; o II, com 2 doses a intervalo 30 dias e reforço aos 210 dias; e, o III com uma dose e reforço aos 210 dias. A revacinação anual foi realizada em todos os animais aos 365 dias. As amostras de soros foram obtidas aos 0, 30, 60, 90, 150, 210, 240, 300, 365, 395, 545 e 730 dias, e armazenadas à temperatura de -20ºC, e a dosagem dos anticorpos realizada através do teste simplificado da inibição da fluorescência. Verificou-se que os animais do grupo I apresentaram soroconversão mais duradoura (12232 dias) do que os dos grupos II e III (p<0,05) após a vacinação inicial, e os animais do grupo I apresentaram soroconversão mais duradoura (183,6120,6 dias) do que os dos grupos II (p<0,05), após a revacinação anual aos 365 dias. A VARCCL induziu a resposta imune nos macacos-prego, após vacinação e revacinação, respectivamente, em 81,4 por cento e 76,0 por cento dos animais; com produção de anticorpos neutralizantes, iguais ou superiores a 0,5 UI/ml, porém, de curta duração; não constituindo assim, imunógeno apropriado para ser utilizado na rotina de imunização destes animais de difícil lide, mantidos em cativeiro / Twenty-seven capuchin monkeys (Cebus apella) were intramuscularly immunized with inactivated suckling mouse brain rabies vaccine (SMBV) employed in campaigns for animal rabies prevention in dogs and cats. The animals were kept confined from June, 1995 to June, 1997. They were divided into 3 experimental groups and vaccinated according to the following scheme: group I, with 3 doses within a 30-day interval; group II, with 2 doses within a 30-day interval and a booster dose at the 210th day; and, group III; with a single dose and a booster dose at the 210th day. All animals were given the annual re-vaccination at the 365th day. Sera samples were obtained at the 0th, 30 th, 60th, 90 th, 150 th, 210 th, 240 th, 300 th, 365 th, 395 th, 545 th e 730 th days and kept stored at 20ºC. The antibodies dosage was carried out through the simplified inhibition fluorescent test. The following results were observed: animals belonging to group I had longer humoral immune response (12232 days) than the ones belonging either to group II or group III (p<0.05) after initial vaccination; animals of group I presented longer humoral immune response (183.6120.6 days) than the ones of the group II (p<0.05) after the annual re-vaccination at the 365th day. The SMBV induced humoral immune response in capuchin monkeys after vaccination and re-vaccination in respectively 81.4 per cent and 76.0 per cent of the animals, producing neutralizing antibodies equal to or higher than 0.5 IU/ml; however, they were short-lasting, being therefore not appropriate as an immunogen to be used routinely in the immunization of these animals which are difficult both to be deal with and to be held confined

Equine arteritis virus (EAV) is the causal agent of equine viral arteritis, a disease of equids. During natural outbreaks of the disease, EAV can cause abortion in pregnant mares and persistent infection in stallions. Understanding how host cellular proteins interact with viral RNA and viral proteins, as well as their role in viral infection, will enable better characterization of the pathogenesis of EAV and establishment of persistent infection in stallions. Accordingly, we hypothesized that both viral factors and host genetically related factors could influence the outcome of EAV infection in horses. To test this hypothesis, we first combined contemporary molecular biology techniques with dual color flow cytometric analysis to characterize the interactions of viral structural proteins and the equine peripheral blood mononuclear cells in vitro. Results from this study demonstrated that interactions between GP2, GP3, GP4, GP5 and M envelope proteins of EAV play a major role in determining the CD14+ monocyte tropism while the tropism of CD3+ T lymphocytes is determined by GP2, GP4, GP5 and M envelope proteins but not the GP3 protein. Secondly, a genome wide association study using SNP genotyping identified a common haplotype associated with the in vitro CD3+ T lymphocyte/resistance to EAV infection among four breeds of horses. Subsequently, these studies were extended to establish a possible correlation between the in vitro susceptibility of CD3+ T lymphocytes to EAV and establishment of persistent infection in stallions. Interestingly, carrier stallions with susceptible CD3+ T lymphocyte phenotype to EAV may represent those at higher risk of becoming persistently infected. Finally, the precise effect of EAV on the immune system of horses, innate and humoral immunity, was studied. Horses were shown to mount a strong humoral antibody response to nonstructural proteins (nsps) 2, 4, 5 and 12 of EAV, whereas nsps 1, 2 and 11 suppressed the type I interferon production. The data presented in this dissertation suggest new directions for future EAV research using genomic and proteomic approaches to study host cell factors involved in EAV attachment and entry and establishment of persistent infection in the stallions.

Quatre études épidémiologiques menées en Afrique ont montré que les enfants issus de mères ayant un placenta infecté par Plasmodium falciparum lors de l’accouchement font des infections palustres plus précocement que les autres enfants. Le fœtus serait sensibilisé in utero par les parasites infectants et développerait par la suite une tolérance aux infections palustres. Cette hypothèse nous conduit à supposer que i) la réponse anticorps spécifique de P. falciparum est différente chez les enfants en fonction du statut infectieux du placenta des mères à l’accouchement et ii) que cette sensibilité ne pourrait être induite que par les antigènes parasitaires porteurs des mêmes polymorphismes que ceux rencontrés in utero. Un troisième projet a consisté au développement d’une méthodologie permettant de distinguer les anticorps maternels de ceux néo-synthétisés par l’enfant dans le but de mesurer précisément l’acquisition de la réponse anticorps élaborée par l’enfant dès son plus jeune âge. Nous avons mis en place le suivi régulier et rapproché d’une cohorte de 620 nouveau-nés de la naissance à 18 mois au Bénin. Nous avons mesuré leurs réponses anticorps dirigées contre sept antigènes de P. falciparum candidats vaccins et constaté que le processus de maturation immunitaire commence à être mis en place à l’âge de 18 mois. L’infection palustre placentaire ne semble pas influer sur l’acquisition de la réponse anticorps spécifique jusqu’à 18 mois de vie. La méthodologie de distinction des anticorps maternels et néo-synthétisés a été validée. La caractérisation des polymorphismes des antigènes parasitaires présents à l’accouchement et pendant le suivi des enfants, mis en relation avec les données environnementales, a permis de valider en partie l’hypothèse de tolérance immunitaire. / Four epidemiological studies showed that infants born from mothers with Plasmodium falciparum placental malaria at delivery present a higher susceptibility to plasmodial infections than others. In connection with this observation, we hypothesized that i) the infants’ P. falciparum specific antibody responses are different according to presence or absence of placental malaria at delivery in their mothers and ii) susceptibility could only be induced by antigens that bring the same polymorphisms as those found in infected mothers. Another project consisted to develop a new methodology to distinguish maternal and neonatal antibodies in order to measure accurately neo-synthesized antibodies in the first months of life. A birth cohort of 620 newborns was established in an area endemic for malaria. Infants were followed-up until 18 months of age and their antibody responses specific for 7 P. falciparum antigens were quarterly measured. The emergence of the immune maturation process was observed in 18-months-infants. The acquisition of specific antibody responses was not impacted by placental malaria. The new methodological approach leading to distinguish maternal and neonatal antibodies was validated. The genetic characterization of the parasite antigen polymorphisms in mothers at delivery and their infants during the follow-up, in link to environmental data, led partially to the validation of the immune tolerance hypothesis.

Orientador: Gervásio Henrique Bechara / Banca: Hélio José Montassier / Banca: Osvaldo Augusto Brasil Esteves Sant'Anna / Resumo: No hospedeiro bovino o nível de resistência ao carrapato Rhipicephalus (Boophilus) microplus varia de acordo com a raça sendo os fenótipos contrastantes herdáveis. O presente trabalho explora esses fenótipos a fim de estabelecer os perfis das respostas imunes, humoral e inflamatória, correlacionados com resistência em raça zebuína (Nelore) e susceptibilidade em raça taurina (HPB). Os animais foram expostos a infestação natural pelo R.(B.) microplus e amostras de soros foram coletadas em pontos estratégicos da cinética das infestações. Os níveis de imunoglobulinas totais (IgG1 e IgG2), bem como os de anticorpos IgG1, IgG2 e IgE anti-extrato de ovo, anti-extrato de larva não alimentada e anti-saliva foram determinados. Elementos da resposta inflamatória, como as principais proteínas de fase aguda e óxido nítrico, também foram dosados. Os resultados obtidos mostram que as infestações muito intensas, em hospedeiros suscetíveis, são capazes de modular os níveis séricos de IgG1 e IgG2 total, diminuindo-os significativamente em relação aos níveis observados durantes infestações menores. Também modulam negativamente a produção de todos os anticorpos específicos IgG1 e IgG2 avaliados a nível sistêmico. Bovinos susceptíveis ao carrapato produzem níveis mais altos de anticorpos IgE para todos os antígenos. O fenótipo susceptivel de infestação se diferencia pela maior freqüência do alótipo de IgG?2a, herdado por herança Mendeliana co-dominante. Animais suscetíveis, quando infestados, produzem níveis mais altos da proteína de fase aguda a1- glicoproteína ácida, de padrão anti-inflamatório, enquanto que animais resistentes produziram relativamente mais proteínas de fase aguda pró-inflamatórias, haptoglobina e amilóide sérica A. Em ambas as raças não houve diferença nos níveis de transferrina e óxido nítrico sistêmico, porém a produção de ambos é influenciada pelos níveis de infestação. / Abstract: In bovine hosts resistance to the cattle tick, Rhipicephalus (Boophilus) microplus, varies according to the breed, being the phenotypes of infestations inherited. The present work exploits these contrasting phenotypes in order to determine the profile of the humoral, inflammatory and acute phase responses that are correlated with resistance seen in a zebuine breed (Nelore) and susceptibility seen in a taurine breed (Holstein). Bovines were exposed to natural infestations with R.(B.) microplus and they presented, as expected, different levels of infestation that also varied in intensity according to the season of the year. Samples of sera were collected at strategic points during the kinetics of different cycles of infestations. The levels of total serum IgG1 and IgG2 immunoglobulins, as well as those of IgG1, IgG2 and IgE anti-egg extracts, anti-unfed larvae extracts and anti-saliva antibodies were measured. Components of the inflammatory response, nitric oxide, as well as acute phase proteins, were also measured. The results show that very intense infestations in ticksusceptible bovines modulate the serum levels of IgG1 and IgG2, which are significantly diminished relative to those observed during less intense infestations. Intense infestations also modulate the production of all specific IgG1 and IgG2 antibodies. Susceptible animals produced more specific IgE, suggesting that this isotype does not participate in resistance against ticks. The susceptible cattle also have a higher frequency of IgG?2a , which are encoded by Mendelian co-dominant alleles. When infested susceptible animals produced higher levels of the anti-inflammatory acute phase protein, a1-acid glycoprotein, whereas resistant animals produced relatively higher levels of the pro-inflammatory proteins, haptoglobin and serum amyloide A. / Mestre

Orientador: Iveraldo dos Santos Dutra / Banca: Júlio Cesar de Freitas / Banca: Tereza Cristina Cardoso da Silva / Banca: Raul José Silva Girio / Banca: Samir Issa Samara / Resumo: Foi avaliado pelo teste de ELISA indireto a resposta humoral para as toxinas botulínicas tipos C e D em animais vacinados com quatro diferentes produtos comerciais. Empregou-se duas vacinas bivalentes C e D (vacina 1 e vacina 2) e duas polivalentes contendo ainda os toxóides botulínicos tipos C e D (vacina 3 e vacina 4). Para análise foram realizadas seis colheitas de sangue ao longo do experimento, nos dias 0, 42, 75, 160, 250 e 342 após a vacinação. A imunidade passiva em bezerros até os 90 dias de idade, filhos de mães vacinadas com diferentes toxóides comerciais foram avaliados em 75 bezerros, agrupados de acordo com a vacina utilizada na mãe (B1, B2, B3 e B4). Para análise foram realizadas 3 colheitas de sangue, nos dias 5 (± 2), 45 e 90 dias após o nascimento. A avaliação da resposta humoral de bovinos vacinados em diferentes faixas etárias também foi realizada empregando-se uma vacina antibotulínica bivalente (C e D) comercial. Para análise, 90 bovinos foram divididos em 3 grupos de acordo com a faixa etária; animais com idade inferior a 2 anos, entre 2 e 5 anos e superior a 5 anos. Na avaliação da resposta humoral de animais vacinados com quatro diferentes produtos comerciais, para a toxina tipo C, as vacinas 1, 2, 3 e 4 não apresentaram diferenças significativas aos 42 dias da primeira dose. Aos 75 dias, a vacina 1, foi superior às vacinas 3 e 4, induzindo maior produção de anticorpos nos animais, porém não diferiu da vacina 2. Aos 160 dias houve queda na quantidade de anticorpos em todos os grupos, não havendo mais diferenças significativas entre as quatro vacinas testadas. Para a toxina tipo D, aos 42 dias da vacinação, não houve diferenças significativas entre as vacinas 1, 2 e 4, que apresentaram neste momento, valores superiores a vacina 3. No entanto, quando avaliadas aos 75 dias, a vacina 1 apresentou maior produção de anticorpos, diferindo... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: To compare the efficiency of an "in house" ELISA test, standardized to measure antibody C and D from Clostridium botulinum, a survey to analysis four different commercial vaccines was conducted. The vaccines used were two commercial, bivalent botulinum containing subtypes C and D (vaccine 1 and vaccine 2) and other two polyvalent including subtypes C and D. The first trial was blood samples taken at 0, 42, 75, 160, 250 and 342 days after vaccination, whereas the booster was performed at day 42. The second trial, maternal antibodies were also measured taken blood samples from 1-3 months age steers obtained from vaccinated heifers with the same vaccines described above, and divided into four different groups (B1, B2, B3, B4). The blood samples were taken at days 5, 45 and 90 after birth. Third and last trial was humoral response from vaccinated cattle with different age evaluation performed using the commercial Clostridium botulinum vaccine subtype C and D. For this purpose, 90 animals never vaccinated, were divided into 3 groups: 1 - 2 years old; 2 - animals aged between 2 and 5 years old; 3 - animals 5 years old. The blood samples were taken 30 days after vaccination after second dose of the vaccine. From the first trial, no significant difference was observed when subtype C was search in sera from animals at 42 days after vaccination. However, at 75 days after vaccination, vaccine 1 was able to induce higher levels of antibodies when compared to vaccine 3 and 4. The antibody level was declining after 160 days post vaccination. For subtype D antigen, at 42 days after vaccination, no differences could be observed. However, at 75 day after vaccination, higher levels of antibodies were observed from animals vaccinated with vaccine 1. Comparing the bivalent vaccines, these were able to induce higher antibodies levels at 75 days after vaccination (toxoid C). In addition, the same... (Complete abstract click electronic access below) / Doutor