RESUMO
Concerning the specificities of a longitudinal study, the trajectories of a subject's mean responses not always present a linear behavior, which calls for tools that take into account the non-linearity of individual trajectories and that describe them towards associating possible random effects with each individual. Generalized additive mixed models (GAMMs) have come to solve this problem, since, in this class of models, it is possible to assign specific random effects to individuals, in addition to rewriting the linear term by summing unknown smooth functions, not parametrically specified, then using the P-splines smoothing technique. Thus, this article aims to introduce this methodology applied to a dataset referring to an experiment involving 57 Swiss mice infected by Trypanosoma cruzi, which had their weights monitored for 12 weeks. The analyses showed significant differences in the weight trajectory of the individuals by treatment group; besides, the assumptions required to validate the model were met. Therefore, it is possible to conclude that this methodology is satisfactory in modeling data of longitudinal sort, because, with this approach, in addition to the possibility of including fixed and random effects, these models allow adding complex correlation structures to residuals.
Assuntos
Animais , Masculino , Camundongos , Trypanosoma cruzi/imunologia , Trypanosoma cruzi/parasitologia , Bioterápicos/antagonistas & inibidores , Soro/imunologia , Soro/parasitologia , Trajetória do Peso do Corpo , Pesos e Medidas Corporais , Anticorpos Antiprotozoários/imunologia , Galinhas , Doença de Chagas/tratamento farmacológico , Ensaio Clínico Controlado Aleatório Veterinário , Camundongos , Antígenos de Protozoários/imunologiaRESUMO
Introduction: The antiphospholipid syndrome is characterized by the persistent presence of antiphospholipid antibodies and clinical manifestations of thrombosis or gestational morbidity that are associated with oxidative stress and endothelial dysfunction. Objective: To evaluate markers of oxidative stress in endothelial cells induced by the serum from women with different clinical manifestations of the antiphospholipid syndrome, and to analyze the antioxidant capacity of the sera. Materials and methods: We included 48 women who were classified as follows: presence of antiphospholipid antibodies and clinical criteria of gestational morbidity alone, vascular thrombosis only, and gestational morbidity/vascular thrombosis. Control groups included antiphospholipid antibodies negative women. In an in vitro model of endothelial cells stimulated with sera from women included in the groups, some markers of oxidative stress were determined by flow cytometry. The antioxidant capacity in the sera of these women was analyzed. Results: The sera from the groups of women with antiphospholipid syndrome that presented thrombosis, with or without gestational morbidity, generated a significant increase (p<0.05 and p<0.001) in endothelial oxidative stress markers in contrast to the control of normal human serum. There were no differences in the effect of the sera from the different study groups on endothelial lipid peroxidation. Also, there was also no difference in the antioxidant activity of the sera. Conclusion: Mitochondrial oxidative stress in the endothelium is associated with the presence of thrombosis; instead, its association with gestational morbidity generates intracellular oxidative stress.
Introducción. El síndrome antifosfolípido se caracteriza por la presencia persistente de anticuerpos antifosfolípidos y manifestaciones clínicas de trombosis o morbilidad gestacional, las cuales se asocian con estrés oxidativo y disfunción endotelial. Objetivo. Evaluar los marcadores de estrés oxidativo en células endoteliales, inducidos por el suero de mujeres con diferentes manifestaciones clínicas del síndrome antifosfolípido y analizar la capacidad antioxidante de los sueros. Materiales y métodos. Se incluyeron 48 mujeres que fueron clasificadas así: presencia de anticuerpos antifosfolípidos y criterios clínicos de morbilidad gestacional, trombosis vascular o ambas. Como grupos control se incluyeron mujeres negativas para anticuerpos antifosfolípidos. En un modelo in vitro de células endoteliales estimuladas con los sueros de las mujeres del estudio, se determinaron algunos marcadores de estrés oxidativo por citometría de flujo. También, se analizó la capacidad antioxidante de los sueros incluidos. Resultados. Los sueros de los grupos de mujeres con síndrome antifosfolípido que presentaban trombosis, con morbilidad gestacional o sin ella, generaron un incremento significativo (p<0,05 y p<0,001) en los marcadores de estrés oxidativo endotelial, en contraste con el control de suero humano normal. No se observaron diferencias en el efecto de los sueros de los diferentes grupos de estudio sobre la lipoperoxidación endotelial. Tampoco se encontró diferencia en la actividad antioxidante de los sueros. Conclusión. El estrés oxidativo mitocondrial en el endotelio se asocia con la presencia de trombosis. Sin embargo, cuando esta se asocia con morbilidad gestacional, también se genera estrés oxidativo intracelular.
Assuntos
Anticorpos Antifosfolipídeos/imunologia , Síndrome Antifosfolipídica/imunologia , Células Endoteliais/metabolismo , Estresse Oxidativo/imunologia , Soro/imunologia , Adulto , Síndrome Antifosfolipídica/complicações , Biomarcadores/metabolismo , Estudos de Casos e Controles , Sobrevivência Celular , Feminino , Humanos , Peroxidação de Lipídeos , Gravidez , Complicações na Gravidez/imunologia , Espécies Reativas de Oxigênio/metabolismo , Soro/metabolismo , Superóxidos/metabolismo , Trombose/etiologia , Trombose/imunologia , Veias Umbilicais/citologiaRESUMO
The Costa Rica Vaccine Trial (CVT), a phase III randomized clinical trial, provided the initial data that one dose of the HPV vaccine could provide durable protection against HPV infection. Although the study design was to administer all participants three doses of HPV or control vaccine, 20% of women did not receive the three-dose regimens, mostly due to involuntary reasons unrelated to vaccination. In 2011, we reported that a single dose of the bivalent HPV vaccine could be as efficacious as three doses of the vaccine using the endpoint of persistent HPV infection accumulated over the first four years of the trial; findings independently confirmed in the GSK-sponsored PATRICIA trial. Antibody levels after one dose, although lower than levels elicited by three doses, were 9-times higher than levels elicited by natural infection. Importantly, levels remained essentially constant over at least seven years, suggesting that the observed protection provided by a single dose might be durable. Much work has been done to assure these non-randomized findings are valid. Yet, the group of recipients who received one dose of the bivalent HPV vaccine in the CVT and PATRICIA trials was small and not randomly selected nor blinded to the number of doses received. The next phase of research is to conduct a formal randomized, controlled trial to evaluate the protection afforded by a single dose of HPV vaccine. Complementary studies are in progress to bridge our findings to other populations, and to further document the long-term durability of antibody response following a single dose.
Assuntos
Anticorpos Antivirais/imunologia , Papillomavirus Humano 16/imunologia , Papillomavirus Humano 18/imunologia , Infecções por Papillomavirus/prevenção & controle , Vacinas contra Papillomavirus/administração & dosagem , Adolescente , Adulto , Ensaios Clínicos como Assunto , Costa Rica/epidemiologia , Feminino , Humanos , Esquemas de Imunização , Vacinação em Massa , Infecções por Papillomavirus/epidemiologia , Infecções por Papillomavirus/terapia , Vacinas contra Papillomavirus/uso terapêutico , Soro/imunologia , Fatores de Tempo , Neoplasias do Colo do Útero/epidemiologia , Neoplasias do Colo do Útero/prevenção & controle , Neoplasias do Colo do Útero/terapiaRESUMO
Leptospirosis is a widespread zoonosis caused by pathogenic Leptospira. Bacteria disseminate via the bloodstream and colonize the renal tubules of reservoir hosts. Leptospiral surface-exposed proteins are important targets, because due to their location they can elicit immune response and mediate adhesion and invasion processes. LipL46 has been previously reported to be located at the leptospiral outer membrane and recognized by antibodies present in serum of infected hamsters. In this study, we have confirmed the cellular location of this protein by immunofluorescence and FACS. We have cloned and expressed the recombinant protein LipL46 in its soluble form. LipL46 was recognized by confirmed leptospirosis human serum, suggesting its expression during infection. Binding screening of LipL46 with extracellular matrix (ECM) and plasma components showed that this protein interacts with plasminogen. The binding is dose-dependent on protein concentration, but saturation was not reached with the range of protein concentration used. Kringle domains of plasminogen and lysine residues of the recombinant protein are involved in the binding because the lysine analog, amino caproic acid (ACA) almost totally inhibited the reaction. The interaction of LipL46 with plasminogen generates plasmin in the presence of plasminogen activator uPA. Because plasmin generated at the leptospiral surface can degrade ECM molecules and decrease opsonophagocytosis, we tentatively infer that Lip46 has a role in helping the invasion process of pathogenic Leptospira.
Assuntos
Proteínas da Membrana Bacteriana Externa/metabolismo , Leptospira interrogans/genética , Leptospirose/microbiologia , Lipoproteínas/metabolismo , Plasminogênio/metabolismo , Animais , Anticorpos Antibacterianos/imunologia , Proteínas da Membrana Bacteriana Externa/genética , Proteínas da Membrana Bacteriana Externa/imunologia , Matriz Extracelular/imunologia , Feminino , Humanos , Leptospira interrogans/imunologia , Leptospirose/imunologia , Lipoproteínas/genética , Lipoproteínas/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Plasminogênio/genética , Plasminogênio/imunologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Soro/imunologiaRESUMO
BACKGROUND: Chronic spontaneous urticaria (CSU) may be associated with autoimmune thyroid diseases, and the Autologous Serum Skin Test (ASST) is an autoreactivity marker. The thyrotropin (TSH) and TSH receptor (TSHR) could play a role in the pathogenesis of CSU. The aim of this study was to evaluate ASST positivity and TSHR gene expression in healthy skin and ASST wheals in euthyroid women with CSU, with (14 patients) and without (15 patients) Hashimoto's thyroiditis (HT). METHODS: ASST was performed and TSHR gene expression studied in wheals induced by ASST and in healthy skin. RESULTS: ASST presented greater positivity (86% × 40%) and larger diameter (10.8 × 9.6 mm) in the HT group (P < 0.05). TSHR gene expression was higher in the ASST area and healthy skin of HT group (P < 0.01). Positive correlation of antibodies levels with ASST wheal measurements and TSHR gene expression was seen. CONCLUSIONS: Women with CSU and HT presented greater positivity and larger measurements for ASST and higher TSHR expression in the skin, suggesting association between CSU, thyroid autoimmunity, and TSHR.
Assuntos
Doença de Hashimoto/genética , RNA Mensageiro/metabolismo , Receptores da Tireotropina/genética , Urticária/genética , Adulto , Autoanticorpos/sangue , Estudos de Casos e Controles , Doença Crônica , Feminino , Doença de Hashimoto/complicações , Doença de Hashimoto/metabolismo , Humanos , Iodeto Peroxidase/imunologia , Pessoa de Meia-Idade , Soro/imunologia , Testes Cutâneos , Tireotropina/sangue , Urticária/complicações , Urticária/metabolismoRESUMO
Fasciolosis is caused by Fasciola hepatica that affects the bile ducts and liver parenchyma of ruminants, which can result in economic loss. This study aimed to carry out the validity of the commercial kit ELISA® indirect front of the simple fecal sedimentation test used as the standard. 143 samples were collected blood and feces of cattle from Jerome, south of the Espírito Santo. Serum samples were left at -80 °C and used to perform the ELISA kit IDEXX®. All animals to stool examinations were also positive to the ELISA (22) and negative samples to test stool (121), 52 animals reacted positively against the antibody research. The frequency of fasciolosis was 15.4% in the stool examinations and 51.8% by ELISA. The validity was calculated by sensitivity (100%), specificity (57%), positive predictive value (29%) and negative predictive value (100%), and the correlation between the tests calculated using the kappa index of 0.35. The better sensitivity of the ELISA commercial kit should not be a separately evaluated, since the cost benefit and the technical facility must be considered.(AU)
Fasciolose é causada pela Fasciola hepatica, parasito que acomete os ductos biliares e o parênquima hepático dos ruminantes e pode resultar em perdas econômicas. Objetivou-se realizar a validade do kit comercial ELISA® indireto frente ao teste de sedimentação fecal simples utilizado como padrão. Foram coletadas 143 amostras de sangue e fezes de vacas provenientes do Sul do Espírito Santo. As amostras de sangue foram centrifugadas para separação do soro. As amostras de soro foram congeladas a -80ºC e utilizadas para análise com o kit de ELISA IDEXX®. Todos os 22 animais positivos ao exame coproparasitológico também foram positivos ao ELISA e, das 121 amostras negativas ao exame de fezes, 52 reagiram positivamente frente à pesquisa de anticorpos. A frequência de fasciolose foi de 15,4% no exame coproparasitológico e 51,8% pelo ELISA. A validade foi calculada pela sensibilidade (100%), especificidade (57%), valor preditivo positivo (29%) e valor preditivo negativo (100%), sendo considerada medíocre a concordância entre os testes, calculada pelo índice kappa (0,35). A maior sensibilidade obtida para o kit comercial ELISA não deve ser avaliada isoladamente, uma vez que o custo benefício e a facilidade da técnica devem ser considerados.(AU)
Assuntos
Soro/imunologia , Kit de Medicamentos e Insumos Estratégicos , Fasciolíase/economia , Fasciolíase/veterinária , Bovinos/parasitologiaRESUMO
Escherichia coli O157:H7 is a subtype of Shiga toxin-producing E. coli that is associated with haemorrhagic colitis and haemolytic uremic syndrome (HUS). Studies of populations in endemic areas have reported that the presence of specific antibodies against the O157 lipopolysaccharide (LPS) is associated with a lower incidence of diarrhoea and HUS. Phage display and IgG anti-O157 LPS antibodies were used in the present study to select peptide mimotopes of O157 LPS expressed in protein III of the M13 phage. Synthetic peptides (SP) were designed using the derived amino acid sequences obtained from DNA nucleotides of 63 selected phagotopes. The LxP/YP/SxL motif was identified in five of the phagotope amino acid sequences. Antibody responses against the phagotopes and their corresponding SPs were evaluated. SP12, one of the designed SP, induced the production of antibodies against the homologous peptide (1:800) and O157 LPS (1:200). The specificity of anti-SP12 antiserum was confirmed by analyzing its response to SP3, an SP with a different amino acid sequence than that of SP12, as well as against an E. coli LPS different from O157. Competitive studies with SP12 and O157 LPS showed a significant decrease in anti-SP12 and anti-LPS O157 antiserum responses against SP12 and O157 LPS, respectively. Eighteen (82%) of the 22 human serum samples with positive reactivity against E coli O157 LPS reacted with SP12 SP (cut-off >0.4). These results support the idea that SP12 is an immunogenic mimotope of O157 LPS.
Assuntos
Epitopos/química , Epitopos/imunologia , Escherichia coli O157/imunologia , Peptídeos/química , Peptídeos/imunologia , Animais , Anticorpos/imunologia , Escherichia coli O157/metabolismo , Síndrome Hemolítico-Urêmica/imunologia , Síndrome Hemolítico-Urêmica/metabolismo , Humanos , Imunoglobulina G/imunologia , Lipopolissacarídeos/imunologia , Coelhos , Soro/imunologiaRESUMO
Immunogenicity of ToxA and Vibrio parahaemolyticus lysate was evaluated in a double immunostimulation scheme in Pacific red snapper after V. parahaemolyticus infection. Three groups of Pacific red snapper were intraperitonealy (i.p.) injected with phosphate-buffered saline (PBS group), ToxA of V. parahaemolyticus (ToxA-Vp group) or V. parahaemolyticus lysate (lysate-Vp group) (first injection, day 1; second injection, day 7). Fish were subsequently infected with live V. parahaemolyticus. Humoral immune parameters in skin mucus and serum were evaluated on days 1, 7, 8 and 14 days post-immunostimulation and 7 days post-infection. Moreover expression of immune-related genes was quantified by real time PCR in head-kidney leukocytes, spleen, liver, and intestine. The ToxA-Vp-treated group showed a higher anti-protease and catalase activity in skin mucus when compared with the PBS group. Measurements of SOD and CAT activities showed an increment in both activities a day after the second boost with ToxA-Vp or lysate-Vp. Interestingly, IgM levels in mucus and transcripts were enhanced followed the ToxA-Vp treatment even after challenge. Furthermore, IL-1ß was strongly expressed in all analyzed cell or tissues followed ToxA-Vp or Vp-lysate treatments. Finally, SOD and CAT gene expression was up-regulated in fish immunostimulated with either treatment ToxA-Vp or lysate-Vp, mainly after infection in head-kidney leukocytes and intestine. This is the first study where the effects of ToxA from V. parahaemolyticus in the immune system of Pacific red snapper was evaluated. These results suggest that ToxA-Vp would positively affect humoral immune response and up-regulate expression of genes involved in the immune system function; and could help in the control of V. parahaemolyticus infection in Pacific red snapper Lutjanus peru, an economic important fish in Mexico.
Assuntos
Toxinas Bacterianas/toxicidade , Doenças dos Peixes/imunologia , Imunidade Humoral , Perciformes , Vibrioses/veterinária , Animais , Doenças dos Peixes/microbiologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Expressão Gênica , Muco/imunologia , Especificidade de Órgãos , Soro/imunologia , Vibrioses/imunologia , Vibrioses/microbiologia , Vibrio parahaemolyticus/fisiologiaRESUMO
The aim of this study was to determine the seroprevalence of Toxoplasma gondii infection in free-range chickens from Uberlândia, Minas Gerais state, Brazil, and characterize the genotypic and phenotypic features of two isolates of this parasite, considering the importance of these hosts in the epidemiology of toxoplasmosis. Serum samples from 108 free-range chickens were obtained from ten different districts, and submitted to the modified agglutination test (MAT) for the presence of anti-T. gondii antibodies, and brain and heart tissue samples from infected chickens were processed for mouse bioassay. An overall seroprevalence of 71·3% was found and antibody titres ranged from 16 to 4096. After confirmation of seropositivity by mouse bioassay, the determination of the T. gondii genotypes of two isolates was performed by PCR-RFLP, using primers for the following markers: SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, new SAG2, Apico and CS3. These T. gondii isolates, designated TgChBrUD1and TgChBrUD2, were obtained from heart samples of free-range chickens. The TgChBrUD1 isolate belonged to ToxoDB PCR-RFLP genotype 11 and the TgChBrUD2 isolate belonged to ToxoDB PCR-RFLP genotype 6. Both isolates demonstrated high virulence in a rodent model, with the TgChBrUD1 isolate able to induce brain cysts, in accord with its pattern of multiplication rates in human fibroblast culture. Taken together, these results reveal high prevalence of T. gondii infection in free-range chickens throughout Uberlândia, indicating an important degree of oocyst environmental contamination and the existence of considerable risk for T. gondii transmission to humans by consumption of free-range chicken as a food source.
Assuntos
Galinhas/parasitologia , Toxoplasma/classificação , Toxoplasma/isolamento & purificação , Toxoplasmose Animal/epidemiologia , Toxoplasmose Animal/parasitologia , Testes de Aglutinação , Animais , Anticorpos Antiprotozoários/sangue , Bioensaio , Encéfalo/parasitologia , Brasil/epidemiologia , Estudos Transversais , DNA de Protozoário/genética , Genótipo , Coração/parasitologia , Camundongos , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Estudos Soroepidemiológicos , Soro/imunologia , Toxoplasma/genética , Toxoplasma/fisiologia , VirulênciaRESUMO
Human strongyloidiasis is an infection caused by the helminth Strongyloides stercoralis that can be fatal, especially in immunosuppressed patients. The aim of this study is to evaluate parasite-specific IgG and IgA levels using S. venezuelensis third-stage (L3) infective larvae alkaline extract as a heterologous antigen by ELISA in paired serum and saliva samples with improved sensitivity and specificity. Individuals from northern Paraná state, Brazil were divided into three groups: 30 patients copropositive for S. stercoralis (Group I); 30 clinically healthy individuals (Group II); and 30 patients copropositive for other parasites (Group III). The area under ROC curve (AUC), an overall index of diagnostic accuracy, and Kappa index were calculated. Data were analyzed using analysis of variance (ANOVA) followed by a Kruskal-Wallis test. Probability (p) values of <0.05 were regarded as significant. In Group I, IgG was detected in 96.7% serum and in 6.7% saliva samples. IgG was not detected in Group II. In Group III, cross-reactivity was observed for serum IgG in 26.7% and in 6.7% for saliva samples. In Group I, IgA was detected in 76.7% serum and 56.7% saliva samples. In Group II, 3.3% were positive for IgA in serum, whereas IgA was not detected in any saliva samples. Group III showed 6.7% serum and 26.7% saliva-positive samples. The sensitivity values for detection of IgG and IgA in serum samples were 96.7% and 76.7%, respectively. In saliva samples, the sensitivity values for detection of IgG and IgA were 6.7% and 56.7%, respectively. The specificity value was 100% for the detection of IgG in serum and for detection of IgG and IgA in saliva, and 96.7% for detection of IgA in serum samples. The proper choice of immunological diagnosis to supplement parasitological methods is essential to estimate the true prevalence of the parasite, and will permit analysis of population immune response profiles, particularly in northern Paraná state, where there are no previous reports.
Assuntos
Anticorpos Anti-Helmínticos/análise , Strongyloides stercoralis/imunologia , Estrongiloidíase/diagnóstico , Adolescente , Adulto , Idoso , Animais , Anticorpos Anti-Helmínticos/sangue , Área Sob a Curva , Brasil/epidemiologia , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Hospedeiro Imunocomprometido , Imunoglobulina A/análise , Imunoglobulina A/sangue , Imunoglobulina G/análise , Imunoglobulina G/sangue , Larva/imunologia , Masculino , Pessoa de Meia-Idade , Saliva/imunologia , Sensibilidade e Especificidade , Soro/imunologia , Estrongiloidíase/epidemiologia , Estrongiloidíase/parasitologia , Adulto JovemRESUMO
Shigella flexneri is divided into 13 serotypes based on the combination of antigenic determinants present in the O-antigen. A new O-antigen modification with phosphoethanolamine has been identified. The presence of this antigenic determinant (called E1037) is recognized by monoclonal antibody MASF IV-1. Given the increasing incidence of these new variants and the difficulty in supplying the monoclonal antibody to our country, we produced a polyclonal antiserum (AA479) through immunization with a S. flexneri Xv strain. The antiserum specificity was assessed by slide agglutination against isolates from clinical cases and a culture collection representing all Shigella serotypes. The results obtained demonstrated a 100% correlation between AA479 absorbed antiserum and monoclonal antibody MASF IV-1. The availability of AA479 antiserum in every public hospital in Argentina will allow us to identify atypical S. flexneri isolates in order to strengthen Shigella surveillance in our country and to compare with global epidemiological data.
Assuntos
Soro/imunologia , Shigella flexneri/classificação , Shigella flexneri/imunologia , Animais , Coelhos , SorotipagemRESUMO
Over recent years Brazil has played an increasingly active role internationally, the result of its model of integration and its foreign policy directives. The health sector is a valuable and strategic area for Brazilian technical cooperation to achieve various objectives, including its development goals. This article describes the main directives of Brazilian foreign policy, conceptually defining and characterizing South-South Cooperation, illustrated through an analysis of two Brazilian technical cooperation initiatives in healthcare: one in South America, the other in Africa. The study concludes that, irrespective of the interests and power asymmetries existing in South-South Cooperation, the objectives of this cooperation were achieved through the technical work.
Nos últimos anos, o Brasil foi ativo no âmbito internacional, tanto por seu modelo de inserção como pelas diretrizes de política externa. O setor saúde é uma ferramenta valiosa e estratégica utilizada pela cooperação técnica brasileira para lograr seus objetivos de desenvolvimento. Este artigo descreve as principais diretrizes de política externa brasileira, conceitua e caracteriza a Cooperação Sul-Sul, ilustrada mediante análise de duas iniciativas de cooperação técnica em saúde do Brasil: na América do Sul e na África. O estudo conclui que, independentemente dos interesses e das assimetrias de poder que existem na Cooperação Sul-Sul, os objetivos dessa cooperação foram alcançados por meio do trabalho técnico.
Assuntos
Humanos , Masculino , Feminino , Criança , Adolescente , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Surtos de Doenças , Febre do Nilo Ocidental/epidemiologia , Vírus do Nilo Ocidental/isolamento & purificação , Fatores Etários , Anticorpos Antivirais/sangue , Anticorpos Antivirais/líquido cefalorraquidiano , Líquido Cefalorraquidiano/imunologia , Líquido Cefalorraquidiano/virologia , Ensaio de Imunoadsorção Enzimática , Mortalidade , Reação em Cadeia da Polimerase , Reação em Cadeia da Polimerase Via Transcriptase Reversa , RNA Viral/sangue , RNA Viral/líquido cefalorraquidiano , Análise de Sobrevida , Soro/imunologia , Soro/virologia , Tunísia/epidemiologia , Febre do Nilo Ocidental/patologia , Febre do Nilo Ocidental/virologiaRESUMO
Coccidioidomycosis (Valley Fever) represents a serious threat to inhabitants of endemic areas of North America. Despite successful clinical isolations of the fungal etiological agent, Coccidioides spp., the screening of environmental samples has had low effectiveness, mainly because of the poor characterization of Coccidioides ecological niche. We explored Valle de las Palmas, Baja California, Mexico, a highly endemic area near the U.S.-Mexico border, where we previously detected Coccidioides via culture-independent molecular methods. By testing the serum from 40-trapped rodents with ELISA, we detected antibodies against Coccidioides in two species: Peromyscus maniculatus and Neotoma lepida. This study comprises the first report of wild rodent serum tested for coccidioidal antibodies, and sets the basis to analyze this pathogen in its natural environment and explore its potential ecological niche.
Assuntos
Anticorpos Antifúngicos/sangue , Coccidioides/imunologia , Coccidioidomicose/imunologia , Coccidioidomicose/veterinária , Peromyscus , Sigmodontinae , Animais , Ensaio de Imunoadsorção Enzimática , México , Soro/imunologiaRESUMO
The objective of this study was to compare the ability of two commercial enzyme-linked immunosorbent assays (ELISAs) and an in-house fluorescent microbead immunoassay (FMIA) to detect IgG antibodies against porcine reproductive and respiratory syndrome virus (PRRSV) types 1 and 2 in serum and oral fluids from boars infected experimentally. Samples from uninfected control pigs and PRRSV-negative field samples were also used. Serum samples were tested by ELISAs (IDEXX Se, HIPRA Se) and an in-house FMIA-Se for detection of PRRSV types 1 and 2. Oral fluids were tested by ELISAs (IDEXX-SO, IDEXX-OF, HIPRA-OF) for detection of PRRSV types 1 and 2. Among the sera, IDEXX-Se and HIPRA-Se had similar sensitivity and specificity (p>0.05); however, IDEXX-Se detected positive animals earlier than HIPRA-Se (p<0.05). FMIA-Se had the highest false-positive rates in known negative field samples (1/205 for IDEXX-Se, 5/205 for HIPRA-Se, and 37/205 for FMIA-Se; p<0.01). Serum and oral fluid samples had similar detection rates and antibody kinetics using the IDEXX tests. There was a higher detection rate in serum than oral fluid using the HIPRA assays. In this study, the nucleocapsid protein utilized as antigen in the FMIAs yielded a low specificity. IDEXX-Se had the earliest detection and similar sensitivity and specificity to the HIPRA-Se.
Assuntos
Anticorpos Antivirais/análise , Anticorpos Antivirais/sangue , Técnicas de Laboratório Clínico/métodos , Síndrome Respiratória e Reprodutiva Suína/diagnóstico , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Medicina Veterinária/métodos , Animais , Ensaio de Imunoadsorção Enzimática , Imunoensaio/métodos , Saliva/imunologia , Sensibilidade e Especificidade , Soro/imunologia , Sus scrofa , SuínosRESUMO
Paracoccidioidomycosis (PCM) is a systemic mycosis caused by the fungus Paracoccidioides brasiliensis (S1, PS2, and PS3) and by the new species, P. lutzii. Considering that genetic differences in the Paracoccidioides genus could elicit distinct immune responses by the host, current research investigated serum IgG levels to antigens from P. brasiliensis B339 (S1), P. brasiliensis LDR3 (PS2), and atypical strain LDR2 (P. lutzii), in patients with chronic PCM from the northern and west regions of Paraná, Brazil (n = 35). Cell-free antigen (CFA) and high molecular mass fraction (hMM) were produced from each strain. Samples were analyzed by ELISA and immunodiffusion (ID). ELISA positivity using CFA: B339-100 %, LDR3-83 %, and LDR2-74 %. Response to CFA from B339 was more intense (p < 0.05), while there was no difference between LDR3-LDR2. IgG anti-hMM was higher for antigens from B339 or LDR3, when compared with LDR2 (p < 0.05). There was a positive correlation for each strain between CFA-hMM and for hMM between B339-LDR3 and LDR3-LDR2. ID positivity with CFA: B339-63 %, LDR3-66 %, and LDR2-60 %. We conclude that the intensity of reaction of the patients' sera varies with the strain used; hMM influences tests that use CFA, independently of strain; using ID, positive rates were very similar, but there was a large number of false negative results; ELISA tests using antigens from P. brasiliensis S1 were able to detect a larger number of patients than PS2 and P. lutzii (which had a considerable number of false negative results), and therefore, its use may be more appropriate in this region of Brazil.
Assuntos
Anticorpos Antifúngicos/sangue , Antígenos de Fungos , Imunoglobulina G/sangue , Paracoccidioides/imunologia , Paracoccidioidomicose/imunologia , Paracoccidioidomicose/microbiologia , Antígenos de Fungos/imunologia , Brasil , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Paracoccidioidomicose/diagnóstico , Sensibilidade e Especificidade , Soro/imunologiaRESUMO
Serum samples from 169 water buffaloes and 121 beef cattle were analyzed for antibodies to T. gondii by an indirect fluorescent antibody test (IFAT). Positive results were obtained in 27.2% of water buffaloes and 17.4% of cattle. Statistical analysis indicated significant differences between the prevalence in cattle and buffalo (p ≤ 0.05). The highest titres found in positive animals were 1:256 (buffaloes) and 1:64 (cattle). In both bovine species, toxoplasmosis frequency in young animals (less than 2 years old) was lower compared to older individuals, although the differences seen in cattle were not statistically significant.
Assuntos
Anticorpos Antiprotozoários/sangue , Búfalos , Toxoplasma/imunologia , Toxoplasmose Animal/epidemiologia , Animais , Brasil/epidemiologia , Bovinos , Técnica Indireta de Fluorescência para Anticorpo , Estudos Soroepidemiológicos , Soro/imunologiaRESUMO
BACKGROUND: Serological tests have been used for the diagnosis of Taenia solium infection in pigs. However, those serological results do not necessarily correlate with the actual infection burden after performing pig necropsy. This study aimed to evaluate the Electro Immuno Transfer Blot (EITB) seropositivity with infection burden in naturally infected pigs. METHODOLOGY/PRINCIPAL FINDINGS: In an endemic area of Peru, 476 pigs were sampled. Seroprevalence was 60.5 ± 4.5% with a statistically higher proportion of positive older pigs (>8 months) than young pigs. The logistic model showed that pigs >8 month of age were 2.5 times more likely to be EITB-positive than ≤ 8 months. A subset of 84 seropositive pigs were necropsied, with 45.2% (38/84) positive to 1-2 bands, 46.4% (39/84) to 3 bands, and 8.3% (7/84) to 4+ bands. 41 out of 84 positive pigs were negative to necropsy (48.8%) and 43 (51%) had one or more cysts (positive predictive value). Older pigs showed more moderate and heavy infection burdens compared to younger pigs. In general, regardless of the age of the pig, the probability of having more cysts (parasite burden) increases proportionally with the number of EITB bands. CONCLUSIONS/SIGNIFICANCE: The probability of being necropsy-positive increased with the number of bands, and age. Therefore, the EITB is a measure of exposure rather than a test to determine the real prevalence of cysticercosis infection.
Assuntos
Anticorpos Anti-Helmínticos/sangue , Cisticercose/veterinária , Carga Parasitária/métodos , Doenças dos Suínos/imunologia , Doenças dos Suínos/parasitologia , Taenia solium/imunologia , Taenia solium/isolamento & purificação , Animais , Cisticercose/imunologia , Cisticercose/parasitologia , Larva/imunologia , Peru , Soro/imunologia , SuínosRESUMO
The present study aimed to evaluate the effects of two serine proteases from Bothrops pirajai snake venom, named BpirSP27 and BpirSP41, on the complement system and the inflammatory response. The effects of these enzymes on the human complement system were assessed by kinetic hemolytic assays, evaluating the hemolysis promoted by the classical/lectin (CP/LP) and alternative (AP) pathways after incubation of normal human serum with the serine proteases. The results suggested that these enzymes were able to induce modulation of CP/LP and AP at different levels: BpirSP41 showed higher inhibitory effects on the hemolytic activity of CP/LP than BpirSP27, with inhibition values close to 40% and 20%, respectively, for the highest concentration assayed. Regarding AP, both enzymes showed percentages of inhibition of the hemolytic activity around 20% for the highest concentrations tested, indicating similar effects on this complement pathway. The proinflammatory effects of B. pirajai serine proteases were evaluated regarding their ability to induce paw edema, variations in the pain threshold and leukocyte recruitment at the site of injection. Both showed mild effects on these inflammatory processes, leading to low levels of increase of paw volumes and decrease in pain thresholds in rats up to 6 h after injection, and inducing neutrophil recruitment without significant increases in the total number of leukocytes in the inflammatory exudates after 6 and 24 h of administration into mice peritoneal cavity. These results suggest that serine proteases must present a minor role in the inflammation caused by B. pirajai snake venom.