RESUMO
OBJECTIVE: To contribute to the discussion on the research findings indicating the sexual transmission of American trypanosomiasis and Chagas disease in humans. METHODS: A review of the literature was performed to investigate the routes of transmission of Trypanosoma cruzi parasites and to evaluate the distribution of Chagas disease, which is now found across five continents. RESULTS: The epidemiological profile of American trypanosomiasis, which is still considered a neglected disease of the poor people of Latin America, has changed over time. A family-based study demonstrated that the blood protozoan T. cruzi can be transmitted sexually from infected males and females to naïve mates. CONCLUSIONS: Evidence that Chagas disease can be transmitted sexually, coupled with the migration of individuals with Chagas disease to previously non-endemic countries and increased travel to endemic countries, has implications for public health. Improved screening of blood supplies and prenatal care are required to prevent congenital spread.
Assuntos
Doença de Chagas/transmissão , Doenças Negligenciadas/epidemiologia , Infecções Sexualmente Transmissíveis/transmissão , Doença de Chagas/diagnóstico , Doença de Chagas/parasitologia , Feminino , Humanos , América Latina/epidemiologia , Masculino , Doenças Negligenciadas/parasitologia , Cuidado Pré-Natal/organização & administração , Pesquisa , Infecções Sexualmente Transmissíveis/parasitologia , ViagemRESUMO
The capacity of the liver to regenerate is an important clinical issue after major hepatectomies and makes the difference between life and death in some cases of post-operative malfunction when the liver remnant is too small or has an impaired regenerative capacity. Several approaches have been tested to stimulate hepatic regeneration after post-operative hepatic failure syndrome; however, they have produced controversial results. A quick, simple, and harmless method that can be used intraoperatively and capable of promoting an increased regenerative capacity of the remaining liver would be very welcome. Thus, based on the data in the literature, we presented low-power laser irradiation (LPLI) as a quick, simple, and harmless method to improve liver regeneration after major hepatectomies. This article highlights the current evidence about the effects of LPLI on liver regeneration, and also suggests laser therapy as an important tool for regenerative stimulation in clinical practice.
RESUMO
BACKGROUND: Infection with the protozoan Trypanosoma cruzi manifests in mammals as Chagas heart disease. The treatment available for chagasic cardiomyopathy is unsatisfactory. METHODS/PRINCIPAL FINDINGS: To study the disease pathology and its inhibition, we employed a syngeneic chicken model refractory to T. cruzi in which chickens hatched from T. cruzi inoculated eggs retained parasite kDNA (1.4 kb) minicircles. Southern blotting with EcoRI genomic DNA digests revealed main 18 and 20 kb bands by hybridization with a radiolabeled minicircle sequence. Breeding these chickens generated kDNA-mutated F1, F2, and F3 progeny. A targeted-primer TAIL-PCR (tpTAIL-PCR) technique was employed to detect the kDNA integrations. Histocompatible reporter heart grafts were used to detect ongoing inflammatory cardiomyopathy in kDNA-mutated chickens. Fluorochromes were used to label bone marrow CD3+, CD28+, and CD45+ precursors of the thymus-dependent CD8α+ and CD8ß+ effector cells that expressed TCRγδ, vß1 and vß2 receptors, which infiltrated the adult hearts and the reporter heart grafts. CONCLUSIONS/SIGNIFICANCE: Genome modifications in kDNA-mutated chickens can be associated with disruption of immune tolerance to compatible heart grafts and with rejection of the adult host's heart and reporter graft, as well as tissue destruction by effector lymphocytes. Autoimmune heart rejection was largely observed in chickens with kDNA mutations in retrotransposons and in coding genes with roles in cell structure, metabolism, growth, and differentiation. Moreover, killing the sick kDNA-mutated bone marrow cells with cytostatic and anti-folate drugs and transplanting healthy marrow cells inhibited heart rejection. We report here for the first time that healthy bone marrow cells inhibited heart pathology in kDNA+ chickens and thus prevented the genetically driven clinical manifestations of the disease.
Assuntos
Doenças Autoimunes/prevenção & controle , Transplante de Medula Óssea , Cardiomiopatia Chagásica/prevenção & controle , Doença de Chagas/terapia , Animais , Apoptose , Galinhas/genética , DNA de Cinetoplasto/genética , Rejeição de Enxerto , Imunização , Mutação , Miocárdio/patologia , Trypanosoma cruzi/genética , Trypanosoma cruzi/imunologiaRESUMO
A simple, easy, and safe procedure aiming to improve liver regeneration could be of great clinical benefit in critical situations such as major hepatectomy, trauma, or hemorrhage. Low-power laser irradiation (LPLI) has come into a wide range of use in clinical practice by inducing regeneration in healthy and injured tissues. However, the effect of LPLI on the process of liver regeneration, especially those related to the molecular mechanisms, is not fully understood. Thus, the aim of the present study was to investigate the main molecular mechanisms involved in liver regeneration of partially hepatectomized rats exposed to LPLI. We used Wistar male rats, which had their remaining liver irradiated or not with LPLI (wavelength of 632.8 nm and fluence of 65 mW/cm(2)) for 15 min after a 70% hepatectomy. We subsequently investigated hepatocyte growth factor (HGF), Met, Akt, and Erk 1/2 signaling pathways through protein expression and phosphorylation analyses along with cell proliferation (proliferating cell nuclear antigen (PCNA) and Ki-67) using immunoblotting and histological studies. Our results show that LPLI can improve liver regeneration as shown by increased HGF protein expression and the phosphorylation levels of Met, Akt, and Erk 1/2 accompanied by higher levels of the PCNA and Ki-67 protein in the remnant livers. In summary, our results suggest that LPLI may play a clinical role as a simple, fast, and easy-to-perform strategy in order to enhance the liver regenerative capacity of a small liver remnant after hepatectomy.
Assuntos
Fator de Crescimento de Hepatócito/metabolismo , Regeneração Hepática/efeitos da radiação , Terapia com Luz de Baixa Intensidade , Animais , Hepatectomia , Antígeno Ki-67/metabolismo , Regeneração Hepática/fisiologia , Sistema de Sinalização das MAP Quinases/efeitos da radiação , Masculino , Fosforilação , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-met/metabolismo , Ratos , Ratos Wistar , Transdução de Sinais/efeitos da radiaçãoRESUMO
Human populations are constantly plagued by hematophagous insects' bites, in particular the triatomine insects that are vectors of the Trypanosoma cruzi agent in Chagas disease. The pharmacologically-active molecules present in the salivary glands of hematophagous insects are injected into the human skin to initiate acquisition of blood meals. Sets of vasodilators, anti-platelet aggregators, anti-coagulants, immunogenic polypeptides, anesthetics, odorants, antibiotics, and detoxifying molecules have been disclosed with the aid of proteomics and recombinant cDNA techniques. These molecules can provide insights about the insect-pathogen-host interactions essential for understanding the physiopathology of the insect bite. The data and information presented in this review aim for the development of new drugs to prevent insect bites and the insect-transmitted endemic of Chagas disease.
Assuntos
Hemostasia/efeitos dos fármacos , Mordeduras e Picadas de Insetos/fisiopatologia , Proteínas e Peptídeos Salivares/farmacologia , Animais , Apirase/farmacologia , Doença de Chagas/transmissão , Interações Hospedeiro-Patógeno , Humanos , Glândulas Salivares/química , Proteínas e Peptídeos Salivares/química , Triatoma/genética , Vasodilatadores/farmacologiaRESUMO
BACKGROUND: Reactivation of chronic Chagas disease, which occurs in approximately 20% of patients coinfected with HIV/Trypanosoma cruzi (T. cruzi), is commonly characterized by severe meningoencephalitis and myocarditis. The use of quantitative molecular tests to monitor Chagas disease reactivation was analyzed. METHODOLOGY: Polymerase chain reaction (PCR) of kDNA sequences, competitive (C-) PCR and real-time quantitative (q) PCR were compared with blood cultures and xenodiagnosis in samples from 91 patients (57 patients with chronic Chagas disease and 34 with HIV/T. cruzi coinfection), of whom 5 had reactivation of Chagas disease and 29 did not. PRINCIPAL FINDINGS: qRT-PCR showed significant differences between groups; the highest parasitemia was observed in patients infected with HIV/T. cruzi with Chagas disease reactivation (median 1428.90 T. cruzi/mL), followed by patients with HIV/T. cruzi infection without reactivation (median 1.57 T. cruzi/mL) and patients with Chagas disease without HIV (median 0.00 T. cruzi/mL). Spearman's correlation coefficient showed that xenodiagnosis was correlated with blood culture, C-PCR and qRT-PCR. A stronger Spearman correlation index was found between C-PCR and qRT-PCR, the number of parasites and the HIV viral load, expressed as the number of CD4(+) cells or the CD4(+)/CD8(+) ratio. CONCLUSIONS: qRT-PCR distinguished the groups of HIV/T. cruzi coinfected patients with and without reactivation. Therefore, this new method of qRT-PCR is proposed as a tool for prospective studies to analyze the importance of parasitemia (persistent and/or increased) as a criterion for recommending pre-emptive therapy in patients with chronic Chagas disease with HIV infection or immunosuppression. As seen in this study, an increase in HIV viral load and decreases in the number of CD4(+) cells/mm(3) and the CD4(+)/CD8(+) ratio were identified as cofactors for increased parasitemia that can be used to target the introduction of early, pre-emptive therapy.
Assuntos
Doença de Chagas/complicações , Coinfecção/diagnóstico , Infecções por HIV/complicações , HIV/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Trypanosoma cruzi/isolamento & purificação , Carga Viral , Adolescente , Adulto , Contagem de Linfócito CD4 , Relação CD4-CD8 , Doença de Chagas/parasitologia , Coinfecção/parasitologia , Coinfecção/virologia , Feminino , HIV/genética , Infecções por HIV/imunologia , Infecções por HIV/virologia , Humanos , Masculino , Pessoa de Meia-Idade , Trypanosoma cruzi/genética , Adulto JovemRESUMO
Acute Trypanosoma cruzi infections can be asymptomatic, but chronically infected individuals can die of Chagas' disease. The transfer of the parasite mitochondrial kinetoplast DNA (kDNA) minicircle to the genome of chagasic patients can explain the pathogenesis of the disease; in cases of Chagas' disease with evident cardiomyopathy, the kDNA minicircles integrate mainly into retrotransposons at several chromosomes, but the minicircles are also detected in coding regions of genes that regulate cell growth, differentiation, and immune responses. An accurate evaluation of the role played by the genotype alterations in the autoimmune rejection of self-tissues in Chagas' disease is achieved with the cross-kingdom chicken model system, which is refractory to T. cruzi infections. The inoculation of T. cruzi into embryonated eggs prior to incubation generates parasite-free chicks, which retain the kDNA minicircle sequence mainly in the macrochromosome coding genes. Crossbreeding transfers the kDNA mutations to the chicken progeny. The kDNA-mutated chickens develop severe cardiomyopathy in adult life and die of heart failure. The phenotyping of the lesions revealed that cytotoxic CD45, CD8(+) γδ, and CD8α(+) T lymphocytes carry out the rejection of the chicken heart. These results suggest that the inflammatory cardiomyopathy of Chagas' disease is a genetically driven autoimmune disease.
Assuntos
Doença de Chagas/imunologia , Doença de Chagas/parasitologia , Trypanosoma cruzi/fisiologia , Animais , Autoimunidade/imunologia , Galinhas , Modelos Animais de Doenças , Humanos , Trypanosoma cruzi/imunologiaRESUMO
The triatomines in the tribe Rhodniini are the main vectors of the Trypanosoma cruzi to humans in recent outbreaks of acute Chagas disease in the Amazon. These insects dwelling in palm trees do not colonize the human domicile. Their success to transmit the infection relies partially on the efficacy of their salivary gland apparatuses. Here we show the transcriptome of the Rhodnius brethesi and Rhodnius robustus salivary glands, comprising 56 and 122 clusters, respectively. Approximately one third of these clusters are described for the first time. The LC-MS/MS analysis identified 123 and 111 proteins in R. brethesi and R. robustus sialome, respectively. Noteworthy, lipocalin platelet aggregation inhibitors, inositol polyphosphate 5-phosphatases, and Kazal domain proteins, which are essential for the insect's successful acquisition of blood meals, were found in our analysis. Moreover, glutathione S transferase and antigen-5, which play roles in the insect's defense and resistance against insecticide, were also observed.
Assuntos
Insetos Vetores/química , Rhodnius/química , Glândulas Salivares/química , Venenos de Vespas , Animais , Doença de Chagas/parasitologia , Doença de Chagas/transmissão , Perfilação da Expressão Gênica , Glutationa Transferase/análise , Glutationa Transferase/genética , Humanos , Inositol Polifosfato 5-Fosfatases , Proteínas de Insetos/análise , Insetos Vetores/patogenicidade , Resistência a Inseticidas/genética , Lipocalinas/análise , Lipocalinas/genética , Monoéster Fosfórico Hidrolases/análise , Monoéster Fosfórico Hidrolases/genética , Inibidores da Agregação Plaquetária/análise , Rhodnius/parasitologia , Glândulas Salivares/parasitologia , Venenos de Vespas/análise , Venenos de Vespas/genéticaRESUMO
Panstrongylus megistus, a vector for the Chagas disease parasite Trypanosoma cruzi, is a hematophagous bug widely distributed in South America. This ubiquitous triatomine is known to colonize different wild life habitats. Additionally, P. megistus synanthropy, preying upon mammals, birds, reptiles, and eventually being predators upon insect's hemolymph probably increases its ability to survive after prolonged fasting. It was suspected that the P. megistus mechanisms of adaptation to survival might include a salivary gland complex tool-box with a diversity of pharmacologically active proteins for obtaining blood meals. Herein we describe comprehensive proteome and transcriptome of the P. megistus salivary gland. The proteomic analysis led to the identification of 159 proteins, and the transcriptome revealed 47 complete cDNAs. A diversity of protein functions associated to blood feeding was identified. The most prevalent proteins were related to blood clotting, anti-platelet aggregation and anti-vasoconstriction activities, which correlate with the insect's ability to obtain meals from different sources. Moreover, a gene of resistance to insecticides was identified. These features augments the comprehension towards P. megistus enormous capacity to survive in adverse wild life-changing habitats.
Assuntos
Comportamento Alimentar , Proteínas de Insetos/análise , Panstrongylus/química , Proteínas e Peptídeos Salivares/fisiologia , Animais , Anticoagulantes , Hemolinfa , Proteínas de Insetos/fisiologia , Insetos Vetores , Resistência a Inseticidas , Panstrongylus/parasitologia , Panstrongylus/fisiologia , Inibidores da Agregação Plaquetária , Glândulas Salivares/química , Glândulas Salivares/parasitologia , Triatoma , Trypanosoma cruzi , Vasoconstrição/efeitos dos fármacosRESUMO
BACKGROUND: The administration of anti-trypanosome nitroderivatives curtails Trypanosoma cruzi infection in Chagas disease patients, but does not prevent destructive lesions in the heart. This observation suggests that an effective treatment for the disease requires understanding its pathogenesis. METHODOLOGY/PRINCIPAL FINDINGS: To understand the origin of clinical manifestations of the heart disease we used a chicken model system in which infection can be initiated in the egg, but parasite persistence is precluded. T. cruzi inoculation into the air chamber of embryonated chicken eggs generated chicks that retained only the parasite mitochondrial kinetoplast DNA minicircle in their genome after eight days of gestation. Crossbreeding showed that minicircles were transferred vertically via the germ line to chicken progeny. Minicircle integration in coding regions was shown by targeted-primer thermal asymmetric interlaced PCR, and detected by direct genomic analysis. The kDNA-mutated chickens died with arrhythmias, shortness of breath, cyanosis and heart failure. These chickens with cardiomyopathy had rupture of the dystrophin and other genes that regulate cell growth and differentiation. Tissue pathology revealed inflammatory dilated cardiomegaly whereby immune system mononuclear cells lyse parasite-free target heart fibers. The heart cell destruction implicated a thymus-dependent, autoimmune; self-tissue rejection carried out by CD45(+), CD8γδ(+), and CD8α lymphocytes. CONCLUSIONS/SIGNIFICANCE: These results suggest that genetic alterations resulting from kDNA integration in the host genome lead to autoimmune-mediated destruction of heart tissue in the absence of T. cruzi parasites.
Assuntos
Cardiomiopatia Chagásica/patologia , Modelos Animais de Doenças , Doenças das Aves Domésticas/patologia , Trypanosoma cruzi/patogenicidade , Animais , Doenças Autoimunes/patologia , Antígenos CD8/análise , Galinhas , DNA Circular/genética , DNA Circular/isolamento & purificação , DNA Mitocondrial/genética , DNA Mitocondrial/isolamento & purificação , DNA de Protozoário/genética , DNA de Protozoário/isolamento & purificação , Insuficiência Cardíaca , Interações Hospedeiro-Parasita , Antígenos Comuns de Leucócito/análise , Subpopulações de Linfócitos/química , Subpopulações de Linfócitos/imunologia , Miocardite/patologia , Miocárdio/patologia , Reação em Cadeia da Polimerase/métodos , Trypanosoma cruzi/genéticaRESUMO
The triatomine bugs are obligatory haematophagous organisms that act as vectors of Chagas disease by transmitting the protozoan Trypanosoma cruzi. Their feeding success is strongly related to salivary proteins that allow these insects to access blood by counteracting host haemostatic mechanisms. Proteomic studies were performed on saliva from the Amazonian triatomine bugs: Rhodnius brethesi and R. robustus, species epidemiologically relevant in the transmission of T. cruzi. Initially, salivary proteins were separated by two-dimensional gel electrophoresis (2-DE). The average number of spots of the R. brethesi and R. robustus saliva samples were 129 and 135, respectively. The 2-DE profiles were very similar between the two species. Identification of spots by peptide mass fingerprinting afforded limited efficiency, since very few species-specific salivary protein sequences are available in public sequence databases. Therefore, peptide fragmentation and de novo sequencing using a MALDI-TOF/TOF mass spectrometer were applied for similarity-driven identifications which generated very positive results. The data revealed mainly lipocalin-like proteins which promote blood feeding of these insects. The redundancy of saliva sequence identification suggested multiple isoforms caused by gene duplication followed by gene modification and/or post-translational modifications. In the first experimental assay, these proteins were predominantly phosphorylated, suggesting functional phosphoregulation of the lipocalins.
Assuntos
Doença de Chagas/transmissão , Insetos Vetores/química , Proteômica/métodos , Proteínas e Peptídeos Salivares/análise , Trypanosoma cruzi/química , Animais , Eletroforese em Gel Bidimensional , Comportamento Alimentar , Lipocalinas/análise , Lipocalinas/genética , Proteínas de Protozoários/análise , Proteínas de Protozoários/genética , Rhodnius/química , Rhodnius/parasitologia , Saliva/química , Proteínas e Peptídeos Salivares/genética , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Triatominae/química , Trypanosoma cruzi/patogenicidadeRESUMO
Interspecies DNA transfer is a major biological process leading to the accumulation of mutations inherited by sexual reproduction among eukaryotes. Lateral DNA transfer events and their inheritance has been challenging to document. In this study we modified a thermal asymmetric interlaced PCR by using additional targeted primers, along with Southern blots, fluorescence techniques, and bioinformatics, to identify lateral DNA transfer events from parasite to host. Instances of naturally occurring human infections by Trypanosoma cruzi are documented, where mitochondrial minicircles integrated mainly into retrotransposable LINE-1 of various chromosomes. The founders of five families show minicircle integrations that were transferred vertically to their progeny. Microhomology end-joining of 6 to 22 AC-rich nucleotide repeats in the minicircles and host DNA mediates foreign DNA integration. Heterogeneous minicircle sequences were distributed randomly among families, with diversity increasing due to subsequent rearrangement of inserted fragments. Mosaic recombination and hitchhiking on retrotransposition events to different loci were more prevalent in germ line as compared to somatic cells. Potential new genes, pseudogenes, and knockouts were identified. A pathway of minicircle integration and maintenance in the host genome is suggested. Thus, infection by T. cruzi has the unexpected consequence of increasing human genetic diversity, and Chagas disease may be a fortuitous share of negative selection. This demonstration of contemporary transfer of eukaryotic DNA to the human genome and its subsequent inheritance by descendants introduces a significant change in the scientific concept of evolutionary biology and medicine.
Assuntos
Doença de Chagas/genética , DNA de Protozoário/genética , Transferência Genética Horizontal , Trypanosoma cruzi/genética , Adolescente , Adulto , Idoso , Animais , Brasil , Doença de Chagas/parasitologia , Criança , Feminino , Genoma Humano/genética , Geografia , Interações Hospedeiro-Parasita/genética , Humanos , Hibridização in Situ Fluorescente , Elementos Nucleotídeos Longos e Dispersos/genética , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Linhagem , Recombinação Genética , Análise de Sequência de DNA , Trypanosoma cruzi/fisiologia , Células U937 , Adulto JovemRESUMO
An epidemiological chain involving Trypanosoma cruzi is discussed at the environmental level, and in terms of fine molecular interactions in invertebrate and vertebrate hosts dwelling in different ecosystems. This protozoan has a complex, genetically controlled plasticity, which confers adaptation to approximately 40 blood-sucking triatomine species and to over 1,000 mammalian species, fulfilling diverse metabolic requirements in its complex life-cycle. The Tr. cruzi infections are deeply embedded in countless ecotypes, where they are difficult to defeat using the control methods that are currently available. Many more field and laboratory studies are required to obtain data and information that may be used for the control and prevention of Tr. cruzi infections and their various disease manifestations. Emphasis should be placed on those sensitive interactions at cellular and environmental levels that could become selected targets for disease prevention. In the short term, new technologies for social mobilization should be used by people and organizations working for justice and equality through health information and promotion. A mass media directed program could deliver education, information and communication to protect the inhabitants at risk of contracting Tr. cruzi infections.
Assuntos
Doença de Chagas/parasitologia , Ecossistema , Interações Hospedeiro-Parasita/fisiologia , Insetos Vetores/parasitologia , Triatominae/parasitologia , Trypanosoma cruzi/fisiologia , Animais , Brasil , Doença de Chagas/transmissão , Reservatórios de Doenças/parasitologia , Árvores/parasitologiaRESUMO
An epidemiological chain involving Trypanosoma cruzi is discussed at the environmental level, and in terms of fine molecular interactions in invertebrate and vertebrate hosts dwelling in different ecosystems. This protozoan has a complex, genetically controlled plasticity, which confers adaptation to approximately 40 blood-sucking triatomine species and to over 1,000 mammalian species, fulfilling diverse metabolic requirements in its complex life-cycle. The Tr. cruzi infections are deeply embedded in countless ecotypes, where they are difficult to defeat using the control methods that are currently available. Many more field and laboratory studies are required to obtain data and information that may be used for the control and prevention of Tr. cruzi infections and their various disease manifestations. Emphasis should be placed on those sensitive interactions at cellular and environmental levels that could become selected targets for disease prevention. In the short term, new technologies for social mobilization should be used by people and organizations working for justice and equality through health information and promotion. A mass media directed program could deliver education, information and communication to protect the inhabitants at risk of contracting Tr. cruzi infections.
Uma rede epidemiológica envolvendo o Trypanosoma cruzi foi discutida nos níveis ambientais e de interações moleculares nos hospedeiros que habitam em 19 diferentes ecossistemas. O protozoário tem uma enorme plasticidade controlada geneticamente que confere sua adaptação a cerca de quarenta espécies de triatomíneos e mais de mil espécies de mamíferos. Essas infecções estão profundamente embutidas em inúmeros ecótopos, onde elas estão inacessíveis aos métodos de controle utilizados. Muito mais estudos de campo e de laboratório são necessários à obtenção de dados e informação pertinentes ao controle e prevenção das infecções pelo Tr. cruzi e as várias manifestações da doença. Ênfase deve ser dada àquelas interações que ocorrem nos níveis celulares e ambientais que se poderiam tomar como alvos seletivos para prevenção da doença. Novas tecnologias para mobilização social devem ser disponibilizadas para os que trabalham pela justiça e pela igualdade, mediante informação para a promoção da saúde. Um programa direcionado de educação de massa pode prover informação e comunicação necessárias para proteger os habitantes atualmente expostos ao risco de contrair as infecções pelo Tr. cruzi.
Assuntos
Animais , Doença de Chagas/parasitologia , Ecossistema , Interações Hospedeiro-Parasita/fisiologia , Insetos Vetores/parasitologia , Triatominae/parasitologia , Trypanosoma cruzi/fisiologia , Brasil , Doença de Chagas/transmissão , Reservatórios de Doenças/parasitologia , Árvores/parasitologiaRESUMO
OBJECTIVE: To find the most reliable screening method for Trypanosoma cruzi infection in blood banks. MATERIAL AND METHODS: Epidemiological data, lymphoproliferation assay, parasitological, conventional serological tests: immunofluorescence, haemagglutination, ELISA with epimastigote and trypomastigote antigens and reference serological tests: trypomastigote excreted-secreted antigens (TESA) blot and chemiluminescent ELISA assay with mucine from trypomastigote forms were applied to individuals with inconclusive serology, non-chagasic individuals and chronic chagasic patients. RESULTS: TESA blot had the best performance when used as a single test in all the groups. In the inconclusive group 20.5% of individuals were positive for TESA blot, 23.3% for either lymphoproliferation or TESA blot, and 17.8% for lymphoproliferation only. Positive lymphoproliferation without detectable antibodies was observed in 5.47% of all inconclusive serology cases. Analysis of six parameters (three serological assays, at least one parasitological test, one lymphoproliferation assay and epidemiological data) in the inconclusive group showed that diagnosis of Chagas' disease was probable in 15 patients who were positive by two or more serological tests or for whom three of those six parameters were positive. CONCLUSION: TESA blot is a good confirmatory test for Chagas' disease in the inconclusive group. Although lymphoproliferation suggests the diagnosis of Chagas' disease in the absence of antibodies when associated with a high epidemiological risk of acquiring Chagas' disease, the data from this study and the characteristics of the lymphoproliferation assay (which is both laborious and time-consuming) do not support its use as a confirmatory test in blood-bank screening. However, our findings underscore the need to develop alternative methods that are not based on antibody detection to improve the diagnosis when serological tests are inconclusive.
Assuntos
Bancos de Sangue , Doença de Chagas/diagnóstico , Testes Sorológicos/estatística & dados numéricos , Trypanosoma cruzi , Adulto , Idoso , Animais , Bioensaio/métodos , Brasil , Proliferação de Células , Doença de Chagas/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Humanos , Immunoblotting/métodos , Linfócitos/fisiologia , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Trypanosoma cruzi/imunologia , Adulto JovemRESUMO
INTRODUÇÃO: Em 2007 os autores descreveram a primeira hepatectomia direita por videolaparoscopia realizada no Brasil. Hepatectomia direita ampliada, também conhecida como trisegmentectomia direita, é procedimento altamente complexo e implica em grande retirada do volume hepático. Os autores descrevem a primeira trisegmentectomia direita por videolaparoscopia realizada no Brasil. TÉCNICA: O paciente é colocado em posição supina em decúbito lateral esquerdo. O cirurgião se coloca entre as pernas da paciente. Utilizamos cinco trocartes, três de 12 mm e dois de 5 mm. Devido à embolização prévia da veia porta direita, o hilo hepático não é dissecado. O pedículo portal direito é seccionado com grampeador laparoscópico de carga vascular por meio de acesso intra-hepático, segundo técnica previamente descrita pelos autores. A seguir procede-se a mobilização do fígado direito seguido de dissecção da veia cava retro-hepática e secção da veia hepática direita. Estes passos são realizados sem manobra de Pringle. O fígado é seccionado com combinação de bisturi harmônico e grampeador endoscópico. O pedículo do segmento 4 é seccionado dentro do fígado. O espécime é retirado por meio de incisão supra-púbica e a área cruenta é revista para verificar hemostasia. O procedimento é encerrado e dreno de sistema fechado é posicionado junto à área cruenta. CONCLUSÃO: Trisegmentectomia hepática direita por videolaparoscopia é procedimento factível e seguro e deve ser considerado para pacientes selecionados. Este procedimento deve ser realizado em centros especializados e por cirurgiões com experiência tanto em cirurgia hepática como cirurgia laparoscópica avançada.
BACKGROUND: Laparoscopic right liver trisectionectomy is a very complex procedure and, to our knowledge, there is only one technical description so far in the English literature. The authors describe the first totally laparoscopic right trisectionectomy performed in Brazil. METHOD: Patient is placed in left semi-lateral decubitus position with surgeon standing between patients' legs. Five trocars, three 12 mm and two 5mm, were used. Due to previous right portal vein embolization, hepatic pedicle is not dissected. Intrahepatic access to the main right Glissonian pedicle is achieved with two small incisions: on the right portion of caudate lobe and another in front of the hilum. A vascular stapling device is inserted between these incisions and fired. Right liver is then mobilized and inferior vena cava is dissected. Right hepatic vein is divided with vascular endoscopic stapler. Line of liver transection is marked along the liver surface including segment 4. Glissonian pedicle from segment 4 is divided during liver transection. Liver transection is accomplished with harmonic scalpel and endoscopic stapling device as appropriate. Specimen is extracted through a suprapubic incision and pneumoperitoneum is reestablished. Raw surface area is checked for hemostasia and biliary leakage. One round abdominal drain is left in place. Right hepatic trisectionectomy is then completed. Conclusion: Totally laparoscopic right trisectionectomy is safe and feasible in selected patients and should be considered for patients with benign or malignant liver neoplasms. However, this complex procedure should be performed by surgeons who have both experience in advanced laparoscopic procedures and open hepatic surgery.
RESUMO
BACKGROUND: Trypanosoma cruzi, a flagellate protozoan, is the etiological agent of Chagas disease, a chronic illness that causes irreversible damage to heart and digestive tract in humans. Previous 2-DE analyses of T. cruzi proteome have not focused on basic proteins, possibly because of inherent difficulties for optimizing 2-DE in the alkaline pH range. However, T. cruzi wide pH range 2-DE gels have shown few visible spots in the alkaline region, indicating that the parasite either did not have an appreciable amount of alkaline proteins or that these proteins were underrepresented in the 2-DE gels. RESULTS: Different IEF conditions using 6-11 pH gradient strips were tested for separation of T. cruzi alkaline proteins. The optimized methodology described here was performed using anodic "paper bridge" sample loading supplemented by increased concentration of DTT and Triton X-100 on Multiphor II (GE Healthcare) equipment and an electrode pad embedded in DTT- containing solution near the cathode in order to avoid depletion of reducing agent during IEF. Landmark proteins were identified by peptide mass fingerprinting allowing the production of an epimastigote 2-DE map. Most identified proteins corresponded to metabolic enzymes, especially those related to amino acid metabolism. The optimized 2-DE protocol was applied in combination with the "two-in-one gel" method to verify the relative expression of the identified proteins between samples from epimastigote and trypomastigote life stages. CONCLUSION: High resolution 2-DE gels of T. cruzi life forms were achieved using the optimized methodology and a partial epimastigote alkaline 2-DE map was built. Among 700 protein spots detected, 422 were alkaline with a pI above 7.0. The "two-in-one gel" method simplified the comparative analysis between T. cruzi life stages since it minimized variations in spot migration and silver-stained spot volumes. The comparative data were in agreement with biological traits of T. cruzi life forms and also corroborated previous T. cruzi proteomic studies. For instance, enzymes related to amino acid metabolism and dehydrogenases were more abundant in epimastigote 2-DE gel whilst trans-sialidase and a paraflagellar protein were found specifically in the trypomastigote 2-DE profile.