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1.
J Microbiol Methods ; 217-218: 106874, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38101579

RESUMO

The objective of this study was to validate an indirect enzyme-linked immunoassay (iELISA) using the recombinant proteins, malate dehydrogenase (MDH) and superoxide dismutase (SOD) [CuZn], as antigens and to evaluate its ability to discriminate antibodies produced by vaccination from those induced by infection in the diagnosis of bovine brucellosis. Sera from six groups were evaluated: G1 - culture-positive animals (52 serum samples) (naturally infected); G2 - non-vaccinated animals (28 serum samples) positive in RBT (Rose Bengal test) and 2ME (2-mercaptoethanol test) selected from brucellosis-positive herds; G3 - animals from a brucellosis-free area (32 serum samples); G4 - S19 vaccinated heifers (114 serum samples); G5 - RB51 vaccinated heifers (60 serum samples); G6 - animals inoculated with inactivated Yersinia enterocolitica O:9 (42 serum samples). Diagnostic sensitivity (DSe) and diagnostic specificity (DSp) were estimated using the frequentist approach and the confidence interval (CI) (95%) calculated by the Clopper-Pearson (exact) method. The DSe for iELISA_MDH in the G1 group was 71.7% (CI 95%: 57.6-83.2%) and for the G2 100.0% (CI 95%: 87.7-100.0%), whereas the DSp was 84.4% in the G3 (CI 95%: 67.2-94.7%). For the iELISA_SOD the DSe was estimated 67.3% for the G1 (CI 95%: 52.9-79.7%) and 71.4% for G2 (CI 95%: 51.3-86.8%), while the DSp for G3 was 87.5% (CI 95%: 71.0-96.5%). iELISA_MDH and iELISA_SOD showed potential to be used in the diagnosis of infected animals, increasing the range of serological tests available for the diagnosis of bovine brucellosis, with the advantage of being S-LPS-free. However, none of the tests could differentiate between infection and vaccination.


Assuntos
Brucelose Bovina , Brucelose , Animais , Bovinos , Feminino , Malato Desidrogenase , Ensaio de Imunoadsorção Enzimática/métodos , Brucelose/diagnóstico , Brucelose/veterinária , Testes Sorológicos/métodos , Anticorpos Antibacterianos , Sensibilidade e Especificidade
2.
Antibiotics (Basel) ; 11(12)2022 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-36551488

RESUMO

BACKGROUND: Staphylococcus aureus is one of the most frequently major mastitis pathogens that cause clinical and subclinical mastitis worldwide. Current antimicrobial treatments are usually ineffective, and the commercially available vaccines lack proven effectiveness. The immunological response elicited by the recombinant S. aureus-cure-associated proteins phosphoglycerate kinase (PGK), enolase (ENO), and elongation factor-G (EF-G) in combination with the granulocyte-macrophage colony-stimulating factor (GM-CSF) DNA vaccination was studied in this work. METHODS: Here, twenty-three C57BL/6 mice were divided into four groups and vaccinated with: G1: none (control); G2: GM-CSF DNA plasmid DNA vaccine; G3: the combination of EF-G+ENO+PGK; and G4: the combinations of EF-G+ENO+PGK proteins plus GM-CSF plasmid DNA vaccine. After 44 days, spleen cells were collected for immunophenotyping and lymphocyte proliferation evaluation by flow cytometry upon S. aureus stimulus. RESULTS: Immunization with the three S. aureus recombinant proteins alone resulted in a higher percentage of IL-17A+ cells among CD8+ T central memory cells, as well as the highest intensity of IL-17A production by overall lymphocytes indicating that the contribution of the combined lymphocyte populations is crucial to sustaining a type 3 cell immunity environment. CONCLUSION: The immunization with three S. aureus-cure-associated recombinant proteins triggered type 3 immunity, which is a highly interesting path to pursue an effective bovine S. aureus mastitis vaccine.

3.
Vaccines (Basel) ; 9(8)2021 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-34452024

RESUMO

Staphylococcus aureus mastitis remains a major challenge for dairy farming. Here, 24 mice were immunized and divided into four groups: G1: control; G2: Granulocyte Macrophage Colony-Stimulating Factor (GM-CSF) DNA vaccine; G3: F0F1 ATP synthase subunit α (SAS), succinyl-diaminopimelate (SDD), and cysteinyl-tRNA synthetase (CTS) recombinant proteins; and G4: SAS+SDD+CTS plus GM-CSF DNA vaccine. The lymphocyte subpopulations, and the intracellular interleukin-17A (IL-17A) and interferon-γ production in the draining lymph node cells were immunophenotyped by flow cytometry. The immunophenotyping and lymphocyte proliferation was determined in spleen cells cultured with and without S. aureus stimulus. Immunization with S. aureus recombinant proteins generated memory cells in draining lymph nodes. Immunization with the three recombinant proteins plus GM-CSF DNA led to an increase in the percentage of IL-17A+ cells among overall CD44+ (memory), T CD4+, CD4+ T CD44+ CD27-, γδ TCR, γδ TCR+ CD44+ CD27+, and TCRVγ4+ cells. Vaccination with S. aureus recombinant proteins associated with GM-CSF DNA vaccine downregulated TH2 immunity. Immunization with the three recombinant proteins plus the GM-CSF DNA led to a proliferation of overall memory T, CD4+, and CD4+ TEM cells upon S. aureus stimulus. This approach fostered type 3 immunity, suggesting the development of a protective immune response against S. aureus.

4.
Vet Parasitol Reg Stud Reports ; 23: 100530, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33678384

RESUMO

Trypanosoma cruzi, the etiologic agent of Chagas disease, is widely distributed in the Americas and is transmitted through vectorial, transfusional, and oral routes. This study aimed to evaluate the risk of vectorial transmission of Chagas disease in municipalities located in southern Minas Gerais, Brazil, by analyzing triatomine specimens collected from 2014 to 2020. All 1522 hematophagous triatomines were identified as Panstrongylus megistus, and were subjected to parasitological and molecular examinations. From 2014 to 2016, approximately 10% of insects were positive in the microscopic analysis of intestinal content, and 27% were positive as detected by the quantitative polymerase chain reaction (qPCR) of the same sampling. However, in the last investigated years, an increase in infected triatomines was observed in microscopic analysis (22%) and qPCR methods (41%). This corroborates the findings of acute human Chagas disease cases, which have increased in the study area from a maximum of 2 cases in previous years to 20 cases in 2019, and 17 cases in 2020 through June. Additionally, bloodmeal sources of infected triatomines were investigated; human blood was detected in up to 85.7% of the samples. Moreover, canine blood was also detected in triatomine intestinal content in recent years, reaching 91% of analyzed insects in 2018. Data on bloodmeal sources have demonstrated human-vector contact and have suggested the participation of dogs in the parasite transmission cycle. These results indicate the risk of T. cruzi vectorial transmission in Southern Minas Gerais and São Paulo owing to the boundary between these states. Thus, enhanced surveillance and vector control of Chagas disease are highly recommended in these areas.


Assuntos
Doença de Chagas , Doenças do Cão , Panstrongylus , Animais , Brasil/epidemiologia , Doença de Chagas/epidemiologia , Doença de Chagas/transmissão , Doença de Chagas/veterinária , Doenças do Cão/epidemiologia , Cães , Humanos , Insetos Vetores , Panstrongylus/parasitologia , Trypanosoma cruzi
5.
Microb Pathog ; 147: 104345, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32544523

RESUMO

Brucellosis serodiagnosis is still a challenge and vaccination is the main measure used to control bovine brucellosis, being S19 and RB51 the most currently used vaccines. So, in order to contribute to brucellosis control, a bidimensional (2D) immunoblot-based approach was used to find immunogenic proteins to be used in serodiagnosis, particularly with ability to be employed in DIVA (Differentiating Infected from Vaccinated Animals) strategy. Immunoproteomic profile of Brucella abortus 2308 was analyzed in 2D western blotting using pooled sera from S19 vaccinated animals, RB51 vaccinated animals, B. abortus naturally infected animals and non-vaccinated seronegative animals. Evaluation of the antigens differentially immunoreactive against the groups of sera showed three proteins of particular importance: MDH (malate dehydrogenase) immunoreactive for S19-vaccinated animals, SOD (superoxide dismutase) reactive for infected animals and ABC transporter (multispecies sugar ABC transporter) reactive against sera from vaccinated animals (S19 and RB51). These three proteins were produced in E. coli and tested in an indirect ELISA (I-ELISA). For MDH, comparison between the vaccinated animals (independent of the vaccine used) and the seropositive and seronegative animals in I-ELISA showed significant differences. Data on the I-ELISA using SOD showed that sera from non-vaccinated naturally infected animals exhibited significant difference in comparison with all other groups. Otherwise, sera from vaccinated animals (S19 and RB51) and from non-vaccinated naturally infected animals did not show significant difference in OD values, but they were all significant different from non-vaccinated seronegative animals using ABC transporter as antigen in I-ELISA. In conclusion, together the 2D western blot analysis and the preliminary I-ELISA results suggest that the combined use of MDH and SOD could be successful employed in a LPS-free protein based serodiagnosis approach to detect bovine brucellosis and to discriminate vaccinated from naturally infected animals, in early post-vaccination stages.


Assuntos
Vacina contra Brucelose , Brucelose Bovina , Brucelose , Animais , Anticorpos Antibacterianos , Brucella abortus , Brucelose Bovina/diagnóstico , Brucelose Bovina/prevenção & controle , Bovinos , Escherichia coli , Testes Sorológicos
6.
Rev. bras. anal. clin ; 51(4): 290-295, 2019/12/30.
Artigo em Português | LILACS | ID: biblio-1103988

RESUMO

Os coccídios intestinais compreendem um grupo de protozoários emergentes e oportunistas. As manifestações clínicas causadas podem variar desde diarreia autolimitada até quadros crônicos associados a distúrbios eletrolíticos. Com a pandemia do HIV/ AIDS, essas infecções ganharam notoriedade devido ao quadro agressivo nesses pacientes. Segundo dados da literatura, em pacientes HIV positivos foram encontradas taxas de infecção média de 24%. Dentre os coccídios intestinais, Cryptosporidium sp. possui maior destaque devido ao seu caráter zoonótico. Há registros da recuperação frequente de oocistos em cães e gatos, tendo sido encontrados em 100% dos animais analisados em um hospital veterinário. Além disso, bovinos também podem atuar como reservatórios, com prevalências no Brasil variando de 0,6% a 72,13%. Atualmente, há variadas técnicas para diagnóstico desses coccídios, desde métodos microscópicos que apresentam limitações técnicas até moleculares com elevado custo associado. Além da importância médica, a prevalência de coccidioses intestinais é relevante como um indicador do status socioeconômico de determinada população e da intensidade de transmissão seja ela antroponótica ou zoonótica. Entretanto, ainda são necessárias melhorias no sentido de tornar mais acessíveis novas técnicas diagnósticas, para se detectarem com mais facilidade e confiabilidade os coccídios intestinais.


Intestinal coccidia comprise a group of emerging and opportunistic protozoa. The manifestations caused may range from diarrhea to chronic variables associated with electrolytic disturbances. With an HIV / AIDS pandemic, these infections can be reported because of the aggressive picture in patients. The data of the literature, in HIV positive patients were 24 mm of average of 24%. Among the intestinal coccidia, Cryptosporidium sp. is more prominent due to its zoonotic nature. Recovery records of dogs and cats were found in 100% of treated animals in a veterinary hospital. In addition, cattle may also act as reservoirs, with prevalences in Brazil varying from 0.6% to 72.13%. Currently, there are several methodologies for the diagnosis of these coccidians, from the microscopic methods that present the main techniques for the diagnosis of these coccidians. In addition to the medical importance, the prevalence of intestinal coccidioses is relevant as an indicator of the socioeconomic status of a given population and the intensity of energy is an anthropopathic or zoonotic. However, improvements are still needed to make new diagnostic techniques more accessible, in order to detect intestinal coccidia more easily and reliably.


Assuntos
Parasitos , Infecções Oportunistas , Síndrome de Imunodeficiência Adquirida/diagnóstico , HIV , Coccidiose/diagnóstico , Interações Hospedeiro-Parasita , Criptosporidiose
7.
Future Microbiol ; 13: 429-436, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29125786

RESUMO

AIM: To produce and test recombinant multiepitope proteins as an alternative assay for the serological diagnosis of cryptococcosis. MATERIALS & METHODS: Previously, synthetic peptides were used to detect anti-Cryptococcus antibodies, and in silico analyses showed that the union of peptides would improve the results. Here, the coding sequences of these peptides were assembled into synthetic genes. Four genes have been cloned and expressed in Escherichia coli, producing recombinant multiepitope proteins: proteins A, B, C and D. RESULTS: All constructs yielded good results; however, protein D showed the best results, with a sensitivity of 88.57% and specificity of 100%. CONCLUSION: The multiepitope proteins were shown to be potential antigens for the diagnosis of cryptococcosis in an attempt to detect anti-Cryptococcus antibodies.


Assuntos
Anticorpos Antifúngicos/imunologia , Criptococose/diagnóstico , Cryptococcus/imunologia , Epitopos de Linfócito B/imunologia , Proteínas Recombinantes/imunologia , Sequência de Aminoácidos , Anticorpos Antifúngicos/sangue , Antígenos de Fungos/genética , Antígenos de Fungos/imunologia , Criptococose/sangue , Criptococose/imunologia , Criptococose/microbiologia , Cryptococcus/genética , Ensaio de Imunoadsorção Enzimática , Epitopos de Linfócito B/genética , Escherichia coli/genética , Humanos , Proteínas Recombinantes/genética , Sensibilidade e Especificidade
8.
Vet Parasitol ; 248: 62-67, 2017 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-29173543

RESUMO

Visceral leishmaniasis (VL) is a neglected tropical disease with dogs serving as reservoirs for one of its etiological agents, Leishmania infantum. In Brazil, VL control involves culling of seropositive dogs, among other actions. However, the most employed serological tests lack accuracy, and are not able to detect canine visceral leishmaniasis (CVL) during the early stages of infection. Early detection of CVL is highly desirable in order to shorten the contact time between the infected reservoirs and the vectors. In this study, we investigated the ability of two multiepitope proteins, PQ10 and PQ20, to detect CVL at earlier stages than currently employed methods, including ITS-1 conventional PCR. Using serum samples from naturally infected dogs, we observed that ELISA-PQ10 and ELISA-PQ20 were able to detect Leishmania infection at earlier time points as compared with kDNA PCR-RFLP in anti-IgG and anti-IgM assays. Using sera from experimentally infected dogs, we monitored seroconversion using multiepitope proteins, ELISA-crude antigen, as well as ITS-1 conventional and real-time PCR. While seroconversion was detected by ELISA-crude antigen in 16.6% of the dogs, multiepitope proteins were able to detect seroconversion in more than 80% of them. Moreover, the ability of ELISA-PQ10 and ELISA-PQ20 to detect Leishmania infection at earlier time points as compared with conventional PCR was also confirmed in experimental infection dogs' sera. Immunofluorescence to Babesia canis and Ehrlichia canis did not show cross-reactions with ELISA-PQ10/PQ20 positive samples. Results of real-time PCR and ELISA with multiepitope proteins were very similar, with concordances between 80 and 100%. Furthermore, our findings indicated that PQ10 and PQ20 immunoassays can be related to parasite load. ELISA-PQ10 and ELISA-PQ20 are more sensitive diagnostic tools for early CVL detection as compared with other methods They could potentially be used in screening tests due to easy execution and low costs facilities.


Assuntos
Anticorpos Anti-Idiotípicos/sangue , Anticorpos Antiprotozoários/sangue , Doenças do Cão/diagnóstico , Ensaio de Imunoadsorção Enzimática/veterinária , Leishmania infantum/isolamento & purificação , Leishmaniose Visceral/veterinária , Animais , Brasil , Reações Cruzadas/imunologia , Doenças do Cão/parasitologia , Cães , Diagnóstico Precoce , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/parasitologia , Masculino , Sensibilidade e Especificidade
9.
Curr Protein Pept Sci ; 16(8): 782-90, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25961398

RESUMO

In the present study, the surface plasmon resonance (SPR) and quartz crystal microbalance (QCM) techniques were employed to kinetically evaluate the binding affinity of a new recombinant chimeric protein (CP10) toward anti-Leishmania infantum antibodies for the immunodiagnostics of the visceral leishmaniasis (VL). This chimeric protein was formed by the union in a same artificial coding DNA of ten different peptides, which showed themselves reactive toward positive canine serum for VL. Using the CP10 in enzyme-linked immunosorbent assays (ELISA), it was possible to detect 80% of the asymptomatic infected dogs. After this, SPR and QCM immunosensors were constructed by the covalent immobilization of the CP10 on a self-assembled monolayer (SAM) formed by adsorption of alkanethiol on gold substrates. The thickness (6.80 nm) and the refractive index (1.475) of the protein on the SAM were simultaneously determined through SPR curves measured in different wavelengths (670 and 785 nm). Interactions between the CP10 and its specific IgGs (anti-CP10 antibodies) were characterized by the electrochemical impedance spectroscopy, SPR and QCM techniques. The equilibrium dissociation constant obtained by SPR (K(D) = 8.27 x 10(-10) mol.L(-1)) and QCM (K(D) = 2.42 x 10(- 10) mol.L(-1)) demonstrated high binding affinity of the CP10 toward anti-CP10 antibodies. In this sense, this work quantitatively proves the strong antigenic character of a new recombinant chimeric protein, giving evidence to potential contribution for the use of this protein in programs of control of the VL.


Assuntos
Anticorpos/metabolismo , Leishmaniose Visceral/imunologia , Técnicas de Microbalança de Cristal de Quartzo/métodos , Proteínas Recombinantes de Fusão/metabolismo , Ressonância de Plasmônio de Superfície/métodos , Animais , Antígenos de Protozoários/metabolismo , Espectroscopia Dielétrica , Cães , Proteínas Imobilizadas/metabolismo , Imunoensaio , Cinética , Ligação Proteica , Coelhos , Refratometria , Reprodutibilidade dos Testes
10.
PLoS Negl Trop Dis ; 9(1): e3429, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25569685

RESUMO

BACKGROUND: Visceral leishmaniasis is the most severe form of leishmaniasis. Worldwide, approximately 20% of zoonotic human visceral leishmaniasis is caused by Leishmania infantum, also known as Leishmania chagasi in Latin America. Current diagnostic methods are not accurate enough to identify Leishmania-infected animals and may compromise the effectiveness of disease control. Therefore, we aimed to produce and test two recombinant multiepitope proteins as a means to improve and increase accuracy in the diagnosis of canine visceral leishmaniasis (CVL). METHODOLOGY/PRINCIPAL FINDINGS: Ten antigenic peptides were identified by CVL ELISA in previous work. In the current proposal, the coding sequences of these ten peptides were assembled into a synthetic gene. Furthermore, other twenty peptides were selected from work by our group where good B and T cell epitopes were mapped. The coding sequences of these peptides were also assembled into a synthetic gene. Both genes have been cloned and expressed in Escherichia coli, producing two multiepitope recombinant proteins, PQ10 and PQ20. These antigens have been used in CVL ELISA and were able to identify asymptomatic dogs (80%) more effectively than EIE-LVC kit, produced by Bio-Manguinhos (0%) and DPP kit (10%). Moreover, our recombinant proteins presented an early detection (before PCR) of infected dogs, with positivities ranging from 23% to 65%, depending on the phase of infection in which sera were acquired. CONCLUSIONS/SIGNIFICANCE: Our study shows that ELISA using the multiepitope proteins PQ10 and PQ20 has great potential in early CVL diagnosis. The use of these proteins in other methodologies, such as immunochromatographic tests, could be beneficial mainly for the detection of asymptomatic dogs.


Assuntos
Doenças do Cão/parasitologia , Epitopos , Leishmania infantum , Leishmaniose Visceral/veterinária , Proteínas Recombinantes/metabolismo , Sequência de Aminoácidos , Animais , Antígenos de Protozoários , Doenças do Cão/diagnóstico , Cães , Ensaio de Imunoadsorção Enzimática/veterinária , Leishmaniose Visceral/sangue , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/parasitologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
11.
Future Microbiol ; 9(7): 871-8, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25156376

RESUMO

AIM: To determine the immunoreactivity of synthetic Cryptococcus-derived peptides. MATERIALS & METHODS: A total of 63 B-cell epitopes from previously identified Cryptococcus gattii immunoreactive proteins were synthesized and evaluated as antigens in ELISAs. The peptides were first evaluated for their ability to react against sera from immunocompetent subjects carrying cryptococcal meningitis. Peptides that yielded high sensitivity and specificity in the first test were then retested with sera from individuals with other fungal pathologies for cross-reactivity determination. RESULTS: Six of 63 synthetic peptides were recognized by antibodies in immunoassays, with a specificity of 100%, sensitivity of 78% and low cross-reactivity. CONCLUSION: We successfully determined the immunoreactivity of selected synthetic peptides of C. gattii derived proteins.


Assuntos
Proteínas de Bactérias/imunologia , Cryptococcus gattii/imunologia , Peptídeos/imunologia , Anticorpos Antifúngicos/análise , Anticorpos Antifúngicos/imunologia , Proteínas de Bactérias/genética , Criptococose/imunologia , Criptococose/microbiologia , Cryptococcus gattii/genética , Ensaio de Imunoadsorção Enzimática , Epitopos de Linfócito B/imunologia , Humanos , Peptídeos/síntese química , Peptídeos/química
12.
Acta Trop ; 137: 25-30, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24801885

RESUMO

Visceral leishmaniasis (VL) is a neglected disease and is fatal if untreated. Dogs serve as reservoirs for Leishmania infantum (syn. L. chagasi) due to their susceptibility to infection and high skin parasitism. Therefore, VL control in Brazil involves the elimination of seropositive dogs, among other actions. However, the most frequently used serological tests have limitations regarding sensitivity and specificity. In this study, we have selected three Leishmania antigens (C1, C8 and C9) and have produced them as recombinant proteins using pET-28a-TEV vector and Escherichia coli BL-21 as expression system. When tested in ELISA with human samples, the C9 antigen was the one showing the most promising results, with 68% sensitivity and 78% specificity. When testing canine samples, the C1, C8 and C9 antigens showed a sensitivity range from 70% to 80% and specificity range from 60% to 90%. The C1 antigen presented higher sensitivity (80%) and the C8 antigen presented higher specificity (90%). Due to it, we decided to mix and test C1 and C8 antigens together, resulting in the C18 antigen. The mix also yielded high percentages of detected symptomatic and asymptomatic dogs however it did not improve the performance of the diagnostic. Comparison of our tests with the tests recommended by the Brazilian Ministry of Health revealed that our antigens' sensitivities and the percentage of detected asymptomatic dogs were much higher. Our results suggest that the C1, C8, C18 and C9 recombinant proteins are good antigens to diagnose canine visceral leishmaniasis and could potentially be used in screening tests. To diagnose human visceral leishmaniasis, the C9 antigen presented reasonable results, but more optimization must be performed for this antigen to provide better performance.


Assuntos
Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários , Portador Sadio/veterinária , Doenças do Cão/diagnóstico , Leishmania infantum/imunologia , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/veterinária , Animais , Antígenos de Protozoários/genética , Brasil , Portador Sadio/diagnóstico , Cães , Ensaio de Imunoadsorção Enzimática/métodos , Escherichia coli/genética , Expressão Gênica , Humanos , Leishmania infantum/genética , Proteínas Recombinantes/genética , Sensibilidade e Especificidade , Testes Sorológicos/métodos
13.
Medicina (Ribeiräo Preto) ; 45(3): 310-317, jul.-set. 2012.
Artigo em Português | LILACS | ID: lil-668612

RESUMO

Modelo do estudo: Estudo retrospectivo com análise de dados de prontuário. Objetivo: O presente estudo teve por objetivo verificar a freqüência de obesidade, diabetes mellitus, hipertensão arterial e dislipidemia em um grupo de mulheres climatéricas. Metodologia: Estudo das primeiras pacientes atendidas no Ambulatório do Climatério (ACLI) do Departamento de Ginecologia e Obstetrícia da Faculdade de Medicina de Ribeirão Preto -USP, no período de1983 a 2007. De fevereiro a abril de 2008 foram coletados dados de peso, estatura, Índice de Massa Corporal (IMC), grupo biológico, diagnóstico de hipertensão, diabetes e dislipidemia. Resultados: De 1983 a 2007, 400 pacientes tiveram seguimento no Ambulatório (ACLI), e 272 apresentavam seus prontuários com os dados do presente estudo devidamente registrados. Dessas 272pacientes, foram selecionadas 628 consultas, sendo que, portanto, em média, cada mulher teve trêsretornos. Estas mulheres tinham idade mínima de 29 e máxima de 80 anos, com IMC mediano acimade 25 kg/m2. O diagnóstico de diabetes, hipertensão e dislipidemia foi detectado em, respectivamente:32%, 68% e 54% dos casos.Conclusão: Mulheres climatéricas atendidas em um hospital de nível de atendimento terciário apresentaram um aumento do IMC e da prevalência de doenças crônicas não transmissíveis com o passar dotempo, o que faz urgir um olhar mais atento dos profissionais de saúde a esse grupo populacional.


Study design: Retrospective analysis of medical records.Purpose: This study aimed to determine the prevalence of obesity, diabetes mellitus, hypertension anddyslipidemia in a group of climacteric women.Methods: Study of the first patients treated at the Menopause Clinic (ACLI), Department of Obstetrics andGynecology, School of Medicine of Ribeirão Preto (USP), from 1983 to 2007. Data on weight, height, BMI,biological group, diagnosis of hypertension, diabetes and dyslipidemia was collected from February/2008 until April/2008. Results: From 1983 until 2007, 400 patients were followed up, and 272 had their records registered. Of these 272 patients, 628 were selected queries, and therefore, on average, each woman had three returns. Women over the age of 29 and maximum of 80 years and median BMI above 25kg/m². The prevalence of diabetes, hypertension and dyslipidemia was respectively 32%, 68% and 54%. The prevalence of NCDs and BMI was higher for the later groups. Conclusion: Climacteric women treated at a hospital level care center showed a worsening of the BMI and the prevalence of noncommunicable chronic diseases over time, which is urging a closer look at health professionals in this population group.


Assuntos
Humanos , Feminino , Pessoa de Meia-Idade , Climatério , Diabetes Mellitus , Dislipidemias , Hipertensão , Menopausa , Obesidade
14.
PLoS Negl Trop Dis ; 5(9): e1310, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21931874

RESUMO

BACKGROUND: Visceral leishmaniasis is the most severe form of leishmaniasis. Approximately 20% of zoonotic human visceral leishmaniasis worldwide is caused by Leishmania infantum, which is also known as Leishmania chagasi in Latin America, and disease incidence is increasing in urban and peri-urban areas of the tropics. In this form of disease, dogs are the main reservoirs. Diagnostic methods used to identify Leishmania infected animals are not able to detect all of the infected ones, which can compromise the effectiveness of disease control. Therefore, to contribute to the improvement of diagnostic methods for canine visceral leishmaniasis (CVL), we aimed to identify and test novel antigens using high-throughput analysis. METHODOLOGY/PRINCIPAL FINDINGS: Immunodominant proteins from L. infantum were mapped in silico to predict B cell epitopes, and the 360 predicted peptides were synthesized on cellulose membranes. Immunoassays were used to select the most reactive peptides, which were then investigated with canine sera. Next, the 10 most reactive peptides were synthesized using solid phase peptide synthesis protocol and tested using ELISA. The sensitivity and specificity of these peptides were also compared to the EIE-LVC Bio-Manguinhos kit, which is recommended by the Brazilian Ministry of Health for use in leishmaniasis control programs. The sensitivity and specificity of the selected synthesized peptides was as high as 88.70% and 95.00%, respectively, whereas the EIE-LVC kit had a sensitivity of 13.08% and 100.00% of specificity. Although the tests based on synthetic peptides were able to diagnose up to 94.80% of asymptomatic dogs with leishmaniasis, the EIE-LVC kit failed to detect the disease in any of the infected asymptomatic dogs. CONCLUSIONS/SIGNIFICANCE: Our study shows that ELISA using synthetic peptides is a technique with great potential for diagnosing CVL; furthermore, the use of these peptides in other diagnostic methodologies, such as immunochromatographic tests, could be beneficial to CVL control programs.


Assuntos
Antígenos de Protozoários/análise , Doenças do Cão/parasitologia , Ensaio de Imunoadsorção Enzimática/métodos , Ensaios de Triagem em Larga Escala/métodos , Leishmaniose Visceral/veterinária , Peptídeos/análise , Animais , Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/imunologia , Infecções Assintomáticas , Reservatórios de Doenças/parasitologia , Reservatórios de Doenças/veterinária , Doenças do Cão/diagnóstico , Cães , Ensaios de Triagem em Larga Escala/normas , Leishmaniose Visceral/diagnóstico , Peptídeos/síntese química , Peptídeos/imunologia , Curva ROC , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
15.
Rev. nutr ; 23(4): 535-541, jul.-ago. 2010. tab
Artigo em Português | LILACS | ID: lil-569126

RESUMO

OBJETIVO: O objetivo da pesquisa foi determinar a perda de massa corporal magra em pacientes após cirurgia bariátrica. MÉTODOS: O estudo retrospectivo foi conduzido com 17 prontuários de mulheres obesas submetidas à Derivação Gástrica em Y de Roux com anel de contenção gástrica, incluindo dados obtidos no período pré-operatório imediato e no 1º, 3º, 6º e 12º meses após a cirurgia. Os dados obtidos no prontuário incluíram a idade, medidas de peso, de altura e massa corporal magra e gorda, calculados pela impedância bioelétrica. RESULTADOS: A média de idade das pacientes foi de 43,1, DP=7,7 anos e durante o seguimento houve diminuição significativa do índice de massa corporal [51,2 (40,2-74,1) para 33,7 (24,8-53,4)kg/m²] e da massa corporal gorda [67,5 (51,2-67,4) para 32,1 (16,4-61,9)kg] em 12 meses de seguimento. No primeiro mês após a cirurgia, houve diminuição da massa corporal magra (M=65,3, DP=7,6 para M=59,7, DP=8,1kg), que representou 8,5 por cento em relação aos valores iniciais, sendo que a partir daí, os dados mantiveram-se constantes. CONCLUSÃO: A perda de massa corporal magra pode refletir uma alteração no metabolismo proteico durante o pós-operatório imediato da cirurgia bariátrica, que pode implicar em evolução clínica e nutricional desfavoráveis.


OBJECTIVE: The aim of this study was to determine changes in lean body mass after bariatric surgery. METHODS: This retrospective study reviewed 17 medical records of obese women who underwent banded Roux-en-Y gastric bypass. The medical records contained data collected immediately before and 1, 3, 6 and 12 months after surgery. The data included age, weight, height and lean and fat body mass determined by bioelectrical impedance analysis. RESULTS: The mean age of the patients was 43.1 years (SD=7.7). Body mass index decreased significantly within 12 months of the surgery, going from 51.2 (40.2-74.1) to 33.7 (24.8-53.4)kg/m², as did fat body mass, going from 67.5 (51.2-67.4) to 32.1 (16.4-61.9)kg. In the first month after surgery, lean body mass decreased from M=65.3 (SD=7.6) to M=59.7 (SD=8.1kg), representing a decrease of 8.5 percent. Lean body mass remained constant after this period. CONCLUSION: Loss of lean body mass may indicate a change in protein metabolism immediately after bariatric surgery, which may result in an unfavorable clinical and nutritional course.


Assuntos
Humanos , Feminino , Adulto , Cirurgia Bariátrica/efeitos adversos , Composição Corporal , Redução de Peso
16.
Rev. bras. farmacogn ; 17(3): 455-465, jul.-set. 2007. ilus
Artigo em Português | LILACS | ID: lil-465486

RESUMO

A doença de Chagas, uma zoonose causada por Trypanosoma cruzi, afeta em todo o mundo cerca de 16 a 18 milhões de pessoas. Sua transmissão ocorre através das fezes de triatomíneos, insetos hematófagos conhecidos como barbeiro. Atualmente, a principal forma de transmissão da doença de Chagas em áreas urbanas é por meio de transfusão de sangue contaminado. A violeta de genciana é o único agente que pode ser empregado na quimioprofilaxia de sangue destinado à transfusão. No entanto, existem algumas restrições ao seu uso. A quimioterapia disponível para a doença de Chagas não é eficaz uma vez que as drogas disponíveis nifurtimox e benznidazol são ativas apenas na fase aguda da doença e apresentam sérios efeitos colaterais. Várias substâncias isoladas de plantas foram avaliadas como agentes anti-T. cruzi, objetivando encontrar drogas com menos efeitos colaterais e maior eficácia para a quimioprofilaxia e quimioterapia da doença de Chagas. Nesta revisão são apresentadas as substâncias de origem natural com atividade anti- T. cruzi.


Chagas'disease, a zoonose caused by Trypanosoma cruzi, affects 16-18 million people in the world. The most important mode of transmission of the disease is associated with the feces of several species of triatomine bugs that are strictly hematophagous. Actually, infected blood transfusion is the major mechanism of transmission in urban areas. Gentian violet is the only available prophylactic drug. Despite its effectiveness, there are some restrictions on its use. The available therapy of Chagas' disease is inadequate since the treatment of patients with the drugs nifurtimox and benznidazole presents serious toxic side effects. The search for new trypanocidal compounds to the treatment of Chagas'disease and to use for eliminating T. cruzi from blood, that are more effective and that do not affect red blood cells is the main goal in the prevention of Chagas' disease. In this review a large set of chemicals of plant origin are enumerated and their trypanocidal activity are briefly discussed.


Assuntos
Alcaloides , Doença de Chagas , Flavonoides , Lignanas , Peptídeos , Quinonas , Terpenos , Trypanosoma cruzi
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