Hydroxymethylnitrofurazone (NFOH) decreases parasitaemia, parasitism and tissue lesion caused by infection with the Bolivia Trypanosoma cruzi type I strain in Swiss and C57BL/6 mice

Abstract The chemical hydroxymethylation of the antimicrobial nitrofurazone leads to the prodrug NFOH, also increases the anti-T. cruzi activities (in vitro and in vivo), as well as showed non-genotoxic (Ames and micronucleus assays). In the present study, we assessed the anti-T. cruzi effect of the NFOH In vivo - in acute Swiss and C57Bl/6 experimental Chagas models. The treatment started at 5 days post-infection during 20 consecutive days (orally, once day, 150mg/kg), and the parasitaemia as well as histopathology analysis were performed. In both experimental murine models, NFOH was able to reduce parasitemia blood avoiding parasitic reactivation, during immunosuppression period (dexamethasone 5mg/kg, 14 days), in 100% of the mice, and decrease tissue parasite nests, demonstrating absence of amastigote forms in all organs (100%) analyzed, data similar to benznidazole (BZN). Therefore, the results shown here pointing to the NFOH as promising compound for further preclinical studies, being a high potential drug to effective and safe chemotherapy to Chagas disease.

Factors related to the discontinuation and mortality rates of a cardiac rehabilitation programme in patients with Chagas disease: a 6-year experience in a Brazilian tertiary centre.

OBJECTIVES: To describe the clinical and sociodemographic characteristics of participants as well as discontinuation and mortality rates in a cardiac rehabilitation programme (CRP) tailored to Chagas disease (CD). METHODS: Participants underwent functional capacity, anthropometry and cardiac function evaluations before beginning a CRP. Univariate and multivariate Cox proportional hazards models were performed to investigate the associations between clinical and sociodemographic characteristics at baseline with discontinuation rates and deaths. RESULTS: Forty-two patients were enrolled in the CRP (61.9% men, mean age of 58.1 ± 11.8 years). During a median follow-up period of 10.8 months, 74% discontinued and 14% died while enrolled in CRP. 34% of the patients who discontinued CRP died during follow-up. White race (HR = 0.09; 95% CI 0.01-1.00), right ventricular systolic dysfunction (HR = 10.54; 95% CI 1.24-89.50) and oxygen pulse (HR = 0.69; 95% CI 0.48-0.99) were independently associated with death while enrolled in CRP. Married status (HR = 0.44; 95% CI 0.21-0.95) was independently associated with discontinuation rates from CRP. VO2 peak (HR = 0.85; 95% CI 0.74-0.98) and CRP discontinuation due to CD-related reasons (HR = 8.33; 95% CI 1.91-36.27) were the variables independently associated with death after discontinuation of CRP. CONCLUSION: In this population, sociodemographic aspects and severity of CD were important determinants of CRP discontinuation and mortality.

OBJECTIFS: Décrire les caractéristiques cliniques et sociodémographiques des participants ainsi que les taux d'abandon et de décès dans un programme de réadaptation cardiaque (PRC) adapté à la maladie de Chagas (MC). MÉTHODES: Les participants ont subi des évaluations de la capacité fonctionnelle, de l'anthropométrie et de la fonction cardiaque avant de commencer un PRC. Des modèles de risques proportionnels de Cox univariés et multivariés ont été appliqués pour étudier les associations entre les caractéristiques cliniques et sociodémographiques au départ avec les taux d'abandon et les décès. RÉSULTATS: 42 patients ont été enrôlés dans le PRC (61,9% d'hommes, âge moyen de 58,1 ± 11,8 ans). Au cours d'une période médiane de suivi de 10,8 mois, 74% ont abandonné et 14% sont décédés durant leur enrôlement au PRC. 34% des patients qui ont arrêté le PRC sont décédés au cours du suivi. La race blanche (HR = 0,09; IC95%: 0,01-1,00), le dysfonctionnement systolique ventriculaire droite (HR = 10,54; IC95%: 1,24-89,50) et le pouls d'oxygène (HR = 0,69; IC95%: 0,48-0,99) étaient indépendamment associés avec le décès lors de l'enrôlement au PRC. Le statut marié (HR = 0,44; IC95%: 0,21-0,95) était indépendamment associé aux taux d'abandon de la CRP. Le pic de VO2 (HR = 0,85; IC95%: 0,74-0,98) et l'arrêt du PRC pour des raisons liées à la MC (HR = 8,33; IC95%: 1,91 à 36,27) étaient les variables indépendamment associées au décès après l'arrêt du PRC. CONCLUSION: Dans cette population, les aspects sociodémographiques et la sévérité de la MC étaient des déterminants importants de l'arrêt du PRC et du décès.

Increased serum gastrin in patients with different clinical forms of Chagas disease coinfected with Helicobacter pylori.

Trypanosoma cruzi and Helicobacter pylori (HP) are pathogens that cause chronic diseases and have been associated with hypergastrinemia. The aim of this study was to evaluate the fasting gastrin levels in patients with different clinical forms of Chagas disease (CD), coinfected or not by HP. The enrolled individuals were outpatients attending at the university hospital. HP infection was assessed by serology and 13 C-urea breath test. Fasting serum gastrin concentration was measured by chemiluminescence assay. Gastric endoscopic and histological features were also evaluated. Associations between CD and serum gastrin level were evaluated in a logistical model, adjusting for age, gender and HP status. A total of 113 patients were evaluated (45 with Chagas disease and 68 controls). In the multivariate analysis, increasing serum gastrin levels (OR= 1.02; 95% CI= 1.01-1.12), increasing age (OR= 1.05; 95% CI= 1.02 - 1.09) and HP-positive status (OR = 2.88; 95% CI = 1.10 - 7.51) remained independently associated with CD. The serum gastrin levels were significantly higher in the group of patients with the cardiodigestive form ( P = 0.03) as well as with digestive form ( P = <0.001) of Chagas disease than in the controls. In conclusion, patients with cardiodigestive and digestive clinical forms of CD have increased basal serum gastrin levels in comparison with controls. Moreover, we also demonstrated that H. pylori coinfection contributes to the hypergastrinemia shown in CD.

Estudo de síntese e estabilidades química e enzimática de fármaco dirigido dendrimérico potencialmente ativo em doença de Chagas / Synthesis, chemical and enzymatic stability studies of targeted dendrimer potentially active in Chagas disease

A doença de Chagas representa um problema de saúde pública em muitos países e regiões. O tratamento consiste em fármacos tóxicos, com eficácia discutível, principalmente, na fase crônica da doença. Assim, faz-se necessário o planejamento de novos quimioterápicos, mais seguros e eficazes. Os dendrímeros são novas arquiteturas moleculares formadas por um foco central e ramificações partindo desse foco. Apresentam diversas aplicações biológicas como, por exemplo, atuar como transportadores de fármacos. Face ao exposto, o objetivo deste trabalho foi o estudo de condições para ligar o ácido anacárdico (AA) em derivado dendrimérico com potencial ação na doença de Chagas, o qual tem como foco central o ácido succínico (AS) e ramificações compostas por arginina (Arg) e lisina (Lys). Sabe-se que a cruzaína, uma cisteíno-protease do T. cruzi, catalisa a hidrólise de ligação peptídica entre lisina e arginina. A síntese dos compostos em fase sólida forneceu os derivados brutos: (1) pró-fármaco AA-K-R-NH2 e (2) G.05 AA-K(AS)-R-NH2, que foram purificados e caracterizados por Cromatografia Líquida de Alta Eficiência e espectrometria de massas. Os compostos purificados AA-K-R-NH2 e AA-K(AS)-R-NH2 apresentaram rendimentos de 34% e 47%, com pureza de 88% e 98%, respectivamente. Os resultados dos experimentos enzimáticos utilizando o AA-K-R-NH2 não foram conclusivos. Acredita-se que a baixa solubilidade e/ou baixa concentração podem ter contribuído para tal. Já na estabilidade química em pH 7,4 (que simula pH sanguíneo), pH 1,2 (que simula pH estomacal) e pH 8,5 (que simula pH intestinal), observou-se que o AA-K(AS)-R-NH2 foi estável durante as 24 h de ensaio. Estes últimos resultados são interessantes, pois espera-se que o pró-fármaco dendrimérico alcance o T. cruzi estruturalmente integro, sofrendo hidrólise e liberação do composto ativo no interior do parasita

Chagas disease is a public health problem in many countries and regions. The treatment consists of toxic drugs, with debatable efficacy, mainly, in the chronic phase of the disease. Thus, it is necessary to plan new chemotherapeutics, safer and more effective than those drugs. Dendrimers are new molecular architectures composed by a central focus and branching from that focus. They present several biological applications, such as acting as drug carriers. Thereby, the goal of this work was the study of conditions to bind anacardic acid (AA) in a dendrimeric derivative with potential action in Chagas disease, which was composed by a central focus of succinic acid (AS) and branches of arginine (Arg) and lysine (Lys). Cruzain, a T. cruzi cysteine protease, is known to catalyze the peptide-binding hydrolysis between lysine and arginine. Synthesis of the solid phase compounds provided the crude derivatives: (1) prodrug AA-KR-NH2 and (2) G.05 AA-K(AS)-R-NH2, which were purified and characterized by High Performance Liquid Chromatography (HPLC) and mass spectrometry. The purified AA-K-R-NH2 and AA-K(AS)-R-NH2 compounds showed yields of 34% and 47%, with purity of 88% and 98% respectively. The results of the enzymatic experiments using AA-K-R-NH2 were not conclusive. It is believed that the low solubility and/or low concentration may have contributed for this. On the chemical stability at pH 7.4 (which simulates blood pH), pH 1.2 (which simulates stomach pH) and pH 8.5 (which simulates intestinal pH), it was observed that AA-K(AS)R-NH2 was stable for 24 hours. These latter results are interesting because the dendrimeric prodrug is expected to reach structurally integral T. cruzi, undergoing hydrolysis and release of the active compound within the parasite

Prevalencia de infección por Trypanosoma cruzi en escolares de Salta y Chaco, Argentina / Seroprevalencia of Trypanosoma cruzi infection in school-age children from localities in Salta and Chaco, Argentina / Prevalência de infecção por Trypanosoma cruzi em crianças em idade escolar de Salta e Chaco, na Argentina

El objetivo del trabajo consistió en identificar la seroprevalencia de la infección por Trypanosoma cruzi en niños en edad escolar en localidades de las provincias de Salta y Chaco. Se trabajó en 44 escuelas de la ciudad de Salta, en parajes de San Carlos; en 10 escuelas de La Unión y en 7 escuelas de Taco Pozo. El trabajo tiene un diseño de corte transversal. La toma de muestra se realizó por punción capilar con equipos Serokit, y la confirmación de los casos seropositivos o dudosos por punción venosa, y obtención de suero, para realizar HAI y ELISA. Los casos seropositivos confirmados fueron tratados con Benznidazol® durante 60 días en dosis de 5-7mg/kg/ día. Para el análisis estadístico se elaboraron tablas con frecuencias absolutas y relativas. Se analizaron 17.884 escolares y se detectaron159 seropositivos, siendo la mayor seroprevalencia en la localidad de La Unión y la menor en la ciudad de Salta. Se aplicó tratamiento a 93 niños (71,54 %) de la ciudad y en el ámbito rural se trataron todos los casos. La vía de infección vectorial fue la de mayor predominio en las madres (64,47%). Se concluye que aunque la seroprevalencia fue menor en la ciudad de Salta que en las zonas rurales, es necesario continuar con la vigilancia.

The aim of this work was to identify seroprevalence of Trypanosoma cruzi infection in school-age children who live in localities from Salta and Chaco. This work was conducted in the following schools: 44 located in Salta city, 10 in La Unión, 7 in Taco Pozo, and several in rural spots around San Carlos town. The design was cross-sectional and the samples were taken by capillary punction with Serokit equipment. Seropositive cases were confirmed by HAI and ELISA performed on serum obtained by venous punction. Confirmed seropositive cases were treated with Benznidazol® for 60 days in doses of 5-7 mg/kg/day. Tables with absolute and relative frequencies were made for statistical analysis. It resulted that the number of school-aged children analyzed was 17,884, 159 being seropositive. The highest seroprevalence was detected in La Unión and the lowest in Salta city. Treatment was given to 93 children (71.54%) from Salta city, while every child was treated in rural areas. Infections in mothers was vector-borne mainly (64.47%). It can be concluded that even though seroprevalence was lower in the city of Salta than in rural areas, it is important to continue monitoring for Chagas disease.

O objetivo do trabalho consistiu em identificar a soroprevalência da infecção por Trypanosoma cruzi em crianças em idade escolar em localidades das províncias de Salta e de Chaco. O trabalho foi realizado em 44 escolas da cidade de Salta, em paragens de San Carlos; em 10 escolas de La Unión e em 7 escolas de Taco Pozo. O mesmo tem um desenho de corte transversal. Tomada de amostra: foi realizada por punção capilar com equipamentos Serokit e a confirmação dos casos soropositivos ou duvidoso por punção venosa e obtenção de soro, para realizar HAI e ELISA. Os casos soropositivos confirmados foram tratados com Benznidazole® durante 60 dias em doses de 5-7 mg/kg/dia. Para a análise estatística foram preparadas tabelas com frequências absolutas e relativas. Analisaram-se 17.884 crianças detectando 159 soropositivos, sendo a maior soroprevalência na cidade de La Unión e a menor na cidade de Salta. Aplicou-se o tratamento a 93 crianças, (71,54%) da cidade e na área rural se trataram todos os casos. A via de infecção vetorial foi a de maior predominância nas mães (64,47%). Conclui-se que embora a soroprevalência tenha sido menor na cidade de Salta do que nas áreas rurais, é necessário continuar com a vigilância.

Alterações ao Doppler tecidual em pacientes com a forma aguda da doença de chagas / Alterations to tissue Doppler in patients with acute form of chagas disease

Fundamento: Recentemente tem ocorrido aumento do número de casos agudos de doença de Chagas, principalmente causados por transmissão oral. A maioria dos pacientes mostra boa evolução, apresentando sintomatologia compatívelcom processo infeccioso sistêmico, porém sem alterações cardíacas significativas ao exame físico, eletrocardiograma eecocardiograma transtorácico.Objetivo: Avaliar alterações ecocardiográficas com análise do Doppler tecidual em pacientes com doença de Chagas aguda. Métodos: Foram avaliados pacientes com diagnóstico de doença de Chagas aguda confirmada por exame parasitológico direto. Esses pacientes foram submetidos a exame físico, eletrocardiograma e ecocardiograma transtorácico, sendocomparados com um grupo controle.Resultados: Foram avaliados 12 casos com doença de Chagas aguda e 15 indivíduos no grupo controle. As variáveis que apresentaram diferenças significativas foram: ondas S’ lateral de VE (DCA = 0,09 ± 0,02 m/seg; GC = 0,11 ± 0,02 m/seg; p = 0,024); E’ lateral (DCA = 0,13 ± 0,03 m/seg; GC = 0,18 ± 0,03 m/seg; p = 0,001); E’ septal do VE (DCA = 0,10± 0,03 m/seg; GC = 0,14 ± 0,03 m/seg; p = 0,008), A’ lateral do VE (DCA = 0,08 ± 0,03 m/seg; GC = 0,12 ± 0,01 m/seg;p = 0,003), onda S’ do VD (DCA = 0,12 ± 0,02 m/seg; GC = 0,17 ± 0,02 m/seg; p < 0,001) e TAPSE (DCA = 1,95 ±0,41 cm; GC = 2,37 ± 0,25 cm; p = 0,006). Conclusões: Em pacientes com doença de Chagas aguda, mesmo quando apresentam evolução benigna, podem ocorrer alterações subclínicas detectadas principalmente ao Doppler tecidual. Essas alterações podem ser importantes na avaliação do tratamento da fase aguda e na sua evolução a longo prazo.

Background: Recently there has been an increased number of cases of acute Chagas disease primarily caused by oral transmission. Most patients have a good outcome, presenting symptoms consistent with systemic infectious process, but no significant cardiac abnormalities on physical examination, electrocardiogram and echocardiogram.Objective: To evaluate echocardiographic changes with tissue Doppler analysis in patients with acute Chagas disease.Methods: We evaluated patients with acute Chagas disease confirmed by cytological examination. These patients underwent a physical examination, eletrocardiogram and transthoracic echocardiography, and compared with a control group. Results: We evaluated 12 patients with acute Chagas disease and 15 subjects in the control group. Variables that showed significant diferences were waves S ‘side of LV (DCA = 0.09 ± 0.02m/sec; CG = 0.11 ± 0.02 m/sec; p = 0.024); and ‘side (DCA = 0.13 ± 0.03 m/sec; CG = 0.18 ±0.03 m/sec; p = 0.001); Septal E ‘LV (DCA = 0.10 ± 0.03 m/sec; CG = 0.14 ± 0.03 m/sec; p = 0.008), A’ lateral LV (DCA = 0.08 ± 0.03 m/sec;CG = 0 12 ± 0.01 m/sec; p = 0,003), S wave ‘RV (DCA = 0.12 ± 0.02 m/sec; CG = 0.17 ± 0.02 m/sec; p < 0.001) and TAPSE (DCA = 1,95cm ± 0.41; CG = 2.37 ± 0.25 cm; p = 0.006). Conclusions: In patients with acute Chagas disease, even when present benign, there may be subclinical alterations detected primarilyby tissue Doppler. These changes may be important in the treatment of acute and its long-term evolution.

Preparation, characterization and evaluation of the in vivo trypanocidal activity of ursolic acid-loaded solid dispersion with poloxamer 407 and sodium caprate

Ursolic acid is a promising candidate for treatment of Chagas disease; however it has low aqueous solubility and intestinal absorption, which are both limiting factors for bioavailability. Among the strategies to enhance the solubility and dissolution of lipophilic drugs, solid dispersions are growing in popularity. In this study, we employed a mixture of the surfactants poloxamer 407 with sodium caprate to produce a solid dispersion containing ursolic acid aimed at enhancing both drug dissolution and in vivo trypanocidal activity. Compared to the physical mixture, the solid dispersion presented higher bulk density and smaller particle size. Fourier Transform Infrared Spectroscopy results showed hydrogen bonding intermolecular interactions between drug and poloxamer 407. X-ray diffractometry experiments revealed the conversion of the drug from its crystalline form to a more soluble amorphous structure. Consequently, the solubility of ursolic acid in the solid dispersion was increased and the drug dissolved in a fast and complete manner. Taken together with the oral absorption-enhancing property of sodium caprate, these results explained the increase of the in vivo trypanocidal activity of ursolic acid in solid dispersion, which also proved to be safe by cytotoxicity evaluation using the LLC-MK2 cell line.

O ácido ursólico é um candidato promissor para o tratamento da doença de Chagas, contudo este fármaco possui baixa solubilidade aquosa e limitada absorção intestinal, ambos os fatores limitantes da biodisponibilidade. Entre as estratégias para potencializar a solubilidade e a dissolução de fármacos lipofílicos, as dispersões sólidas estão crescendo em popularidade. Neste estudo, empregamos mistura dos tensoativos, poloxamer 407 e caprato de sódio, para produzir dispersão sólida contendo ácido ursólico, com o objetivo de aumentar tanto a dissolução do fármaco quanto a atividade tripanocida in vivo. Comparada à mistura física, a dispersão sólida apresentou maior densidade e menor tamanho de partícula. Os resultados da análise de espectroscopia no infravermelho com transformada de Fourier mostraram interações intermoleculares do tipo ligações de hidrogênio entre o fármaco e o poloxamer 407. Os experimentos de difratometria de raio-X revelaram a conversão do fármaco de sua forma cristalina para a forma amorfa, mais solúvel. Consequentemente, a solubilidade do ácido ursólico em dispersão sólida foi aumentada e o fármaco dissolveu-se de maneira mais rápida e completa. Em conjunto com as propriedades promotoras de absorção oral do caprato de sódio, estes resultados explicaram o aumento da atividade tripanocida in vivo do ácido ursólico em dispersão sólida, que também se provou segura após avaliação de citotoxicidade empregando a linhagem celular LLC-MK2.

Profile of Trypanosoma cruzi infection in a tropical medicine reference center, Northern Italy.

BACKGROUND: Chagas disease (CD) is endemic in Central and South America, Mexico and even in some areas of the United States. However, cases have been increasingly recorded also in non-endemic countries. The estimated number of infected people in Europe is in a wide range of 14000 to 181000 subjects, mostly resident in Spain, Italy and the United Kingdom. METHODOLOGY/PRINCIPAL FINDINGS: Retrospective, observational study describing the characteristics of patients with CD who attended the Centre for Tropical Diseases (Negrar, Verona, Italy) between 2005 and 2013. All the patients affected by CD underwent chest X-ray, ECG, echocardiography, barium X-ray of the oesophagus and colonic enema. They were classified in the indeterminate, cardiac, digestive or mixed category according to the results of the screening tests. Treatment with benznidazole (or nifurtimox in case of intolerance to the first line therapy) was offered to all patients, excluding the ones with advanced cardiomiopathy, pregnant and lactating women. Patients included were 332 (73.9% women). We classified 68.1% of patients as having Indeterminate Chagas, 11.1% Cardiac Chagas, 18.7% as Digestive Chagas and 2.1% as Mixed Form. Three hundred and twenty-one patients (96.7%) were treated with benznidazole, and most of them (83.2%) completed the treatment. At least one adverse effect was reported by 27.7% of patients, but they were mostly mild. Only a couple of patients received nifurtimox as second line treatment. CONCLUSIONS/SIGNIFICANCE: Our case series represents the largest cohort of T. cruzi infected patients diagnosed and treated in Italy. An improvement of the access to diagnosis and cure is still needed, considering that about 9200 infected people are estimated to live in Italy. In general, there is an urgent need of common guidelines to better classify and manage patients with CD in non-endemic countries.

Atlas iconográfico dos triatomíneos do Brasil (vetores da doença de Chagas) / Iconographic atlas of triatomine Brazil (vectors of Chagas' disease)

O material conta com ilustrações e informações sobre a morfologia dos insetos, tamanho, habitat, ciclo de vida, além de mapas sobre a diversidade e distribuição geográfica das 66 espécies de barbeiros registradas até o momento...

Valorar la asociación de pruebas diagnósticas incruentas para la detección de alteraciones funcionales miocárdicas autonómiocas y endoteliales en la enfermedad de Chagas / To evaluate the association of noninvasive diagnostic tests for the detection of autonomic and endothelial myocardial functional alterations in Chagas disease

Objetivo: Describir la frecuencia de presencia de alteraciones en un conjunto de pruebas diagnósticas no invasivas, de lesiones miocárdicas, anatómicas autonómicas y de función endotelial en pacientes con Chagas en período crónico si patología demostrada (ex indeterminados) y crónicos con patología respecto de controles no chagásicos. Metodología: en un estudio transversal de dos años de duración se incorporaron 288 individuos, 120 controles (78 mujeres); 154 con serología positiva para Chagas (2-3 reacciones), 114 crónicos sin patología demostrada (73 mujeres) y 54 crónicos con patología (40 mujeres). A todos se les realizó señales promediadas (PVT), variabilidad de la frecuencia cardiaca (VFC) por Holter de 24 horas, dispersión del QT, detección serológica de antireceptores muscarínicos (AA) y ultrasonografía de arteria humeral (EE). La valoración estadística se realizó con chi2, Test T o Mann-Whitney, ANOVA...

Infecção natural de Triatoma brasiliensis brasiliensis, Caicó, Rio Grande do Norte, pelo Trypanosoma cruzi e o estudo de peptídeos antimicrobianos em diferentes espécies de triatomíneos (Hemiptera, Heteroptera, Reduviidae) / Natural infection of Triatoma brasiliensis brasiliensis, Caicó, Rio Grande do Norte by trrypanosoma cruzi and the study of antimicrobial peptides in different triatomine species (hemiptera, heteroptera, reduviidae)

Triatoma brasiliensis é um dos vetores da doença de Chagas no Nordeste do Brasil, apresentando altos índices de infecção natural pelo Trypanosoma cruzi. No presente estudo, dois aspectos foram abordados: primeiro a coleta de exemplares de T. b. brasiliensis oriundos de Caicó, RN, para análise de infecção natural por T. cruzi, em períodos diferentes (abril e novembro de 2011). O segundo aspecto abordado está relacionado ao estudo do gene que codifica defensinas como marcador molecular filogenético para a análise de diferentes espécies de triatomíneos, incluindo o complexo T. brasiliensis, além de exemplares de T. b. brasiliensis coletados em Caicó. Em relação à infecção natural de T. b. brasiliensis por T. cruzi observamos um alto índice (86 porcento), mais especificamente na localidade de Penedo. Também foi observado que não houve diferenças significativas entre as expedições realizadas nos períodos de chuva (abril de 2011) e seca (novembro de 2011)Além disso, cinco isolados de T. cruzi foram caracterizados molecularmente por duas metodologias: mini-exon, 24Salpha rDNA e 18S rDNA. Como resultados observamos os dois genótipos (TcI e TcII) circulando em Caicó. Foram obtidas sequências do gene que codifica defensina, onde diferentes isoformas deste gene puderam ser identificadas e caracterizadas nas diferentes espécies de triatomíneos estudadas. A espécie Panstrongylus megistus ficou nos mesmos clados que representantes do gênero Rhodnius. Houve uma clara separação dos clados em relação aos gêneros Triatoma e Rhodnius. Com relação aos membros do complexo T. brasiliensis, não foi possível uma clara distinção entre as espécies pelo fato da molécula de defensina apresentar menos que 500 bp em seu tamanho, além de ser bastante conservada, não sendo portanto um bom marcador para separar grupos muito proximamente relacionados...

Triatoma brasiliensis is one of the main Chagas disease vectors in NortheasternBrazil, presenting a high natural Trypanosoma cruzi infection rate. In the present work, two different aspects were investigated: First, triatomines of the subspecies T. b. brasiliensis were collected in Caicó, RN for analysis of natural infection by T. cruzi in two dissimilar periods (April and November 2011). The second aspect is related touse of defensin encoding genes as a phylogenetic marker for the study of different triatomine species, mainly the T. brasiliensis species complex, including T. b. brasiliensis specimens from Caicó. The results showed a high infection rate (86 percent) of T. b. brasiliensis from Caicó by T. cruzi, especially in insects from Penedo. Nosignificant differences between the rainy period (April 2011) and dried season (November 2011) could be observed. In addition, five T. cruzi isolates were cultivated and characterized by three different molecular methods (mini-exon, 24Salpha rDNA and 18S rDNA). The results showed the presence of TcI and TcII genotypes circulating in Caicó. Defensin encoding genes were amplified and sequenced for different triatomine species, showing three differing forms. Panstrongylus megistus clusteredtogether with representatives of the genus Rhodnius. A separation between the genus Triatoma and Rhodnius could also be detected. However, inside of the T. brasiliensis species complex the taxa could not be well separated. Since thismolecular marker has a size of less than 500 bp and is highly conserved defensinencoding sequences might not be a good marker to separate closely related species and subspecies...

Immunoglobulin M antibodies against CRA and FRA recombinant antigens of Trypanosoma cruzi in chronic chagasic patients.

Previous works of our research group have demonstrated aspects of the humoral immune response of chronic Chagas disease using the cytoplasmatic repetitive antigen (CRA) and the flagellar repetitive antigen (FRA) of Trypanosoma cruzi. The aim of this work was to analyze the presence of specific immunoglobulin M (IgM) antibodies in chronic chagasic patients using these recombinant antigens of T. cruzi. The positivity of IgM in chronic chagasic patients against CRA and FRA antigens was determined by indirect enzyme-linked immunosorbent assay. We reported no statistical significant differences between the levels of IgM for both recombinant antigens and the different chronic clinical forms of Chagas disease. However, a small proportion of chronic chagasic patients analyzed in this study was positive for this antibody isotype. The findings of this study indicate that the IgM antibodies cannot be used to elucidate the differences in the profile of humoral immune response among chronic chagasic patients with different clinical forms using the CRA and FRA recombinant antigens of T. cruzi.

Determining the C-reactive protein level in patients with different clinical forms of chagas disease.

Chagas disease is caused by Trypanosoma cruzi and remains a health problem in the developing countries of South America. The condition leads to cardiac conduction disturbances and chronic heart failure. In this study, 136 individuals were evaluated by the Chagas Disease Study Group of the Hospital de la Universidad Estatal de Campinas in Brazil to determine the relationship between chronic heart failure and the serum C-reactive protein (CRP) level. When patients were stratified according to the different clinical presentations of Chagas disease, it was found that the CRP levels in those with severe heart disease and non-Chagasic cardiopathy were significantly higher than in controls or those with mild heart disease (P< .05), even when participants were stratified by age (i.e. <40 and > or =40 years). There was a direct linear correlation between age and CRP level, such that the older the individual, the higher the CRP level. These data provide further evidence for an association between chronic inflammation and the development of heart failure. Although CRP elevations are not exclusively related to Chagas disease, the CRP level may be a useful marker for the progression of Chagas disease to a more advanced phase.

[Association between the plasma levels of TNF-alpha, IFN-gamma, IL-10, nitric oxide and specific IgG isotypes in the clinical forms of chronic Chagas disease]. / Associação entre os níveis plasmáticos de TNF-alpha, IFN-gamma, IL-10, óxido nítrico e os isotipos de IgG específicos nas formas clínicas da doença de Chagas crônica.

Chagas disease is an important chronic parasitic disease that affects around 9-11 million people in Latin America. A combination of parasite and host-related factors are probably responsible for pathogenesis in the chronic phase of the disease. Among the host-related factors, the immunological response is a parameter of special interest. Our aim here was to evaluate the plasma levels of the cytokines interferon gamma, interleukin 10 and tumor necrosis factor alpha and the immunoglobulins total IgG and its subclasses 3 and 4, by means of ELISA, and the levels of nitric oxide by means of the Griess reaction, among individuals who were seropositive for Trypanosoma cruzi, presenting the cardiac, indeterminate and digestive clinical forms of the disease, and among seronegative individuals. The seropositive individuals produced significantly higher levels of total IgG and IgG-3. Individuals with the digestive form presented higher levels of IgG-4 and interleukin 10. However, these individuals presented lower levels of nitric oxide than the controls did. The results suggest that the higher levels of interleukin 10 observed among individuals with the digestive form may contribute towards the higher levels of the specific IgG-4 that were seen.

Associação entre os níveis plasmáticos de TNF-α, IFN-γ, IL-10, óxido nítrico e os isotipos de IgG específicos nas formas clínicas da doença de Chagas crônica / Association between the plasma levels of TNF-α, IFN-γ, IL-10, nitric oxide and specific IgG isotypes in the clinical forms of chronic Chagas disease

A doença de Chagas é uma importante doença parasitária crônica, que acomete cerca de 9-11 milhões de pessoas na América Latina. Provavelmente, uma combinação de fatores relacionados ao parasito e ao hospedeiro podem ser os responsáveis pela patogênese na fase crônica da doença. Dentre os fatores relacionados ao hospedeiro, a resposta imunológica é um parâmetro de especial interesse. Objetivamos avaliar os níveis plasmáticos das citocinas interferon gama, interleucina 10, fator de necrose tumoral alfa e das imunoglobulinas G total, 3 e 4, por ELISA e do óxido nítrico, pela reação de Griess, entre indivíduos soronegativos e soropositivos para Trypanosoma cruzi, com as formas clínicas cardíaca, indeterminada e digestiva. Os indivíduos soropositivos para Trypanosoma cruzi produziram níveis significativamente mais elevados de imunoglobulinas G total e G3. Indivíduos com a forma digestiva apresentam níveis mais elevados de imunoglobulina G4 e interleucina 10. Entretanto, tais indivíduos apresentaram menores níveis de óxido nítrico do que controles. Os resultados sugerem que os maiores níveis de IL-10 observados nos indivíduos com a forma digestiva poderiam contribuir com os maiores níveis de IgG4 específicos observados.

Chagas disease is an important chronic parasitic disease that affects around 9-11 million people in Latin America. A combination of parasite and host-related factors are probably responsible for pathogenesis in the chronic phase of the disease. Among the host-related factors, the immunological response is a parameter of special interest. Our aim here was to evaluate the plasma levels of the cytokines interferon gamma, interleukin 10 and tumor necrosis factor alpha and the immunoglobulins total IgG and its subclasses 3 and 4, by means of ELISA, and the levels of nitric oxide by means of the Griess reaction, among individuals who were seropositive for Trypanosoma cruzi, presenting the cardiac, indeterminate and digestive clinical forms of the disease, and among seronegative individuals. The seropositive individuals produced significantly higher levels of total IgG and IgG-3. Individuals with the digestive form presented higher levels of IgG-4 and interleukin 10. However, these individuals presented lower levels of nitric oxide than the controls did. The results suggest that the higher levels of interleukin 10 observed among individuals with the digestive form may contribute towards the higher levels of the specific IgG-4 that were seen.

[Part VI. Antiparasitic treatment for Chagas disease]. / Parte VI. Tratamiento antiparasitario de la enfermedad de Chagas.

As expert consensus has been arisen about universal antiparasitic treatment for all patients infected with Trypanosoma cruzi, most important drugs licensed for Chagas disease treatment are reviewed: nifurtimox and benznidazol, their mechanisms of action, doses, treatment schedules, adverse effects and contraindications. Two other drugs used for Chagas disease treatment, for which a Chilean experience may be exhibited, are allopurinol and itraconazole. Indications for treatment of Chagas disease in immunocompetent patients and immunocompromised hosts are detailed. This chapter refers besides to the evaluation and monitoring of antiparasitic therapy in immunocompromised patients, the availability of drugs and includes various forms facsimiles suggested to perform clinical and laboratory follow up of patients that undergo treatment, indicating the prescribed drug, adverse effects and time of follow up.

Guías clínicas de la enfermedad de Chagas 2006: parte VI: tratamiento antiparasitario de la enfermedad de Chagas / Guidelines for the Chagas disease: part VI: antiparasitic treatment for Chagas disease

As expert consensus has been arisen about universal antiparasitic treatment for all patients infected with Trypanosoma cruzi, most important drugs licensed for Chagas disease treatment are reviewed: nifurtimox and benznidazol, their mechanisms of action, doses, treatment schedules, adverse effects and contraindications. Two other drugs used for Chagas disease treatment, for which a Chilean experience may be exhibited, are allopurinol and itraconazole. Indications for treatment of Chagas disease in immunocompetent patients and inmunocompromised hosts are detailed. This chapter refers besides to the evaluation and monitoring of antiparasitic therapy in inmunocompromised patients, the availability of drugs and includes various forms facsímiles suggested to perform clinical and laboratory follow up of patients that undergo treatment, indicating the prescribed drug, adverse effects and time of follow up.

Con el consenso de expertos de que todo paciente infectado con Trypanosoma cruzi debiera recibir tratamiento anti-parasitario, se revisan los principales medicamentos aprobados para la enfermedad de Chagas: nifurtimox y benznidazol, sus mecanismos de acción, dosis, duración del tratamiento, efectos adversos y contraindicaciones. Se mencionan otros dos medicamentos utilizados en el tratamiento, en el que existe alguna experiencia nacional, como son allopurinol e itraconazol. Se revisan las indicaciones de tratamiento de la enfermedad de Chagas en personas inmuno-competentes y las indicaciones de tratamiento en hospederos inmunodeprimidos. Este capítulo finaliza abordando la evaluación y monitorización de la terapia antiparasitaria en inmunodeprimidos, la disponibilidad de medicamentos e incluye facsímiles de formularios sugeridos para realizar el seguimiento clínico y de laboratorio de los pacientes que son sometidos a tratamiento, indicando el fármaco utilizado, los efectos adversos y el tiempo de seguimiento.

[Guidelines for Chagas disease. 2006. Part I. Introduction and epidemiology]. / Guías clínicas de la enfermedad de Chagas. 2006. Parte I. Introducción y epidemiología.

The natural existence of this vectorial zoonosis and the development since 1991 of initiatives in the American Region in order to control Chagas Disease, did necessary to prepare this document to update the knowledge about the disease and to exhibit guidelines for healthcare workers in Chile, as Chagas disease is a public health problem that needs to be attended in an integral way. A definition for Chagas disease and its classification according to the international classification of diseases (ICD 10) are presented. The general importance and disease load are concise and briefly commented, while the vector geographic distribution, the magnitude in Chile, its reservoir, the parasite main characteristics and its transmission mechanisms are reviewed. A definition for acute suspected and confirmed cases is given, and the epidemiological situation in Chile and a brief cost-benefit analysis of Chagas disease are presented.

[Part II. Chagas disease in adults, infancy and adolescence]. / Parte II. Enfermedad de Chagas en el adulto, la infancia y adolescencia.

The different stages of Chagas disease in adults: acute, undetermined or latent and chronic phases are described. This document contains guidelines for etiological diagnosis of Chagas disease and its treatment. In chronic phase, as cardiac and digestive system (esophagus and colon) are affected, symptoms and signs, evolution of the disease, laboratory analysis and treatment are described. The following topics in congenital Chagas disease are boarded: its prevalence in pregnant women, the importance of mother phase of disease, repercussions of the parasite transmission to the fetus, the frequency of transmission, how the infection to the fetus is produced, the importance of chronic infection in consecutive pregnancies, and clinical consequences to the newborn infant including symptoms of congenital disease. Concomitance with human immunodeficiency virus is commented. No vertically transmitted Chagas disease in infancy and adolescents has similar clinical manifestations as in adults. Direct and indirect laboratory tests of infection are described and an algorithm for diagnosis and follow up of vertical transmission of Trypccnosoma cruzi is presented.