Multiple myeloma and Chagas disease: qPCR as a marker for preemptive antiparasitic therapy: a case reports series and review

ABSTRACT Multiple myeloma (MM) associated with Chagas disease is rarely described. This disease and its therapy suppress T cell and macrophage functions and increase regulatory T cell function, allowing the increase of parasitemia and the risk of Chagas Disease Reactivation (CDR). We aimed to analyze the role of conventional (cPCR) and quantitative Polymerase Chain Reaction (qPCR) for prospective monitoring of T. cruzi parasitemia, searching for markers of preemptive antiparasitic therapy in MM patients with Chagas disease. Moreover, we investigated the incidence and management of hematological diseases and CDR both inside and outside the transplant setting in the MEDLINE database. We found 293 studies and included 31 of them. Around 1.9-2.0% of patients with Chagas disease were reported in patients undergoing Stem Cell Transplantation. One case of CDR was described in eight cases of MM and Chagas disease. We monitored nine MM and Chagas disease patients, seven under Autologous Stem Cell Transplantation (ASCT), during 44.56±32.10 months (mean±SD) using parasitological methods, cPCR, and qPCR. From these patients, three had parasitemia. In the first, up to 256 par Eq/mL were detected, starting from 28 months after ASCT. The second patient dropped out and died soon after the detection of 161.0 par Eq/mL. The third patient had a positive blood culture. Benznidazole induced fast negativity in two cases; followed by notably lower levels in one of them. Increased T. cruzi parasitemia was related to the severity of the underlying disease. We recommend parasitemia monitoring by qPCR for early introduction of preemptive antiparasitic therapy to avoid CDR.

Correlation between parasitemia and different complications of malaria – The clinical outlook

Background: Malaria is one of the most severe public health problems worldwide. India contributes a substantial burden of malaria. It can cause several complications. Aims and Objectives: This study was conducted to determine the association of parasitemia of current malaria with age of patients, hemoglobin level, liver function test (LFT), platelet count, previous history of malaria, and relapse/recrudescence of malaria up to 1 year. Materials and Methods: An observational study conducted in hospital settings included thin and thick smear preparation with Leishman’s staining to determine the parasitemia as per the WHO guidelines from the blood samples of 280 malaria parasite dual antigen (MPDA) kit positive patients and their details were taken during study period of 2 years. Hemoglobin, LFT, and platelet count were tested and they were correlated with parasitemia. The previous history of malaria was taken and follow-up was done up to 1 year for relapse/recrudescence and their association with parasitemia in current disease was evaluated. Statistical tool R was used for data analysis. Results: Age group 20–40 years was most commonly affected with maximum mean percentage parasitemia. Higher parasitemia was associated with higher grade of anemia, LFT derangement, and thrombocytopenia. Disease severity was maximum for mixed infection followed by falciparum and vivax malaria. About 9.28% of patients with the previous history of malaria developed lower parasitemia in current infection. Relapse rate in vivax malaria –2.325% and recrudescence rate in falciparum malaria–12.5%. Conclusion: Malaria parasitemia should be reported routinely as it carries prognostic importance.

Sequential analysis of erythrocyte aggregation in P. falciparum malaria with and without ASAQ therapy by optical signal and image analysis

The aggregation of erythrocytes is an important mechanism for blood flow through the cardiovascular system. In malaria, this is complicated by infection caused by P. falciparum and is further complicated by the severity of parasitemia. Hence analysis of this micro-mechanism is essential to know the changes in blood not only in diseased conditions but also after artemisinin combination therapy (ASAQ) to alleviate suffering. For analysis purposes, aggregation of erythrocytes was determined by LED laser aggregometer, represented in terms of various parameters related to the changes in laser transmitted intensity. Formed aggregates are further analyzed by imaging and image-processing methods. For this study blood samples from young adults (18 – 40 years old) infected with P. falciparum (n= 80), without any other serious illness, were performed. These samples were selected based on the severity of parasitemia, and were divided into low (LP), medium1 (MP1), medium 2 (MP2), and high (HP) parasitemia. For three days, the selected individuals were treated with artemisinin-based combination therapy ASAQ (Artesunate 4 mg/kg and amodiaquine 10 mg base/ kg once a day). Healthy subjects (n=20) without any history of the disease were selected as a control group. The results, as obtained by various parameters, show a significant elevation of aggregation of erythrocytes (P< 0.05) in P. falciparum malaria with the increase of parasitemia level. There was a decrease in the aggregation after treatment on day four tending towards normal. Thus the current study shows the potential beneficial role of ASAQ on erythrocytes aggregation, which may contribute to reducing the harmful effects on various organs in P. falciparum-infected blood.

Signos de alarma en pacientes con paludismo importado por Plasmodium falciparum / Warning signs in patients with imported Plasmodium falciparum malaria

Introducción: El paludismo es una enfermedad febril aguda potencialmente mortal causada por parásitos que se transmiten al ser humano por la picadura de mosquitos del género Anopheles. De los 214 millones de casos de paludismo registrados en 2016, la mayoría de ellos se producen en niños menores de cinco años en África subsahariana. La mortalidad está dada por la presencia de sus complicaciones que deben ser detectadas y tratadas precozmente. Objetivo: Identificar la presencia de signos de alarma, y determinar su relación con otras variables clínicas y de laboratorio. Métodos: Se realizó un estudio descriptivo de 47 pacientes adultos con paludismo por Plasmodium falciparum importado, ingresados en el Departamento de Medicina del Instituto de Medicina Tropical Pedro Kourí, desde enero de 2016 a diciembre de 2018. Los datos fueron procesados en una base de datos en Microsoft Excel y luego analizados en el programa estadístico SPSS 11,5. Resultados: Predominaron los pacientes del sexo masculino, con una media de edad de 35,9 años. Fue significativa la relación existente entre los signos de alarma y la severidad del cuadro clínico, la hiperparasitemia, el supuesto estado no inmune de los pacientes, trombocitopenia y la demora en el ingreso. La respuesta al tratamiento es excelente con los esquemas combinados utilizados a base de quinina. Conclusiones: Los signos de alarma, dentro de los cuales podemos incluir la trombocitopenia, constituyen elementos importantes para poder prevenir futuras complicaciones(AU)

Introduction: Malaria is an acute potentially fatal febrile disease caused by parasites transmitted to humans through the bite of mosquitoes from the genus Anopheles. Most of the 214 million malaria cases reported in the year 2016 were children aged under five years from Sub-Saharan Africa. Mortality is due to the presence of complications which should be detected and treated timely. Objective: Identify the presence of warning signs and determine their relationship to other clinical and laboratory variables. Methods: A descriptive study was conducted of 47 adult patients with imported Plasmodium falciparum malaria admitted to the Medicine Department of Pedro Kourí Tropical Medicine Institute from January 2016 to December 2018. The data obtained were processed in a Microsoft Excel database and then analyzed with the statistical software SPSS 11.5. Results: Male patients prevailed, with a mean age of 35.9 years. A significant relationship was found between warning signs and severity of the clinical status, hyperparasitemia, the supposed non-immune status of patients, thrombocytopenia and admission delay. An excellent response was obtained to treatment with combined quinine-based schemes. Conclusions: Warning signs, among them thrombocytopenia, are important to prevent future complications(AU)

Caracterización clínica del paludismo importado / Clinical characterization of imported malaria

Introducción: El paludismo es una enfermedad febril aguda potencialmente mortal causada por parásitos que se transmiten al ser humano por la picadura de mosquitos del género Anopheles. Cuba logró eliminar la transmisión de esta enfermedad gracias a grandes esfuerzos encaminados a conseguirlo, por lo que es necesario adoptar una serie de medidas para evitar su reaparición, mediante la vigilancia y el Programa de Control Sanitario Internacional. Objetivo: Caracterizar clínicamente un grupo de pacientes con paludismo importado. Métodos: Se realizó un estudio descriptivo de corte transversal de 46 pacientes adultos con paludismo importado, ingresados en el Instituto de Medicina Tropical Pedro Kourí desde enero 2015 a diciembre 2016. Los datos fueron tomados de las historias clínicas. El análisis de las variables cualitativas fue expresado en tablas de frecuencias absolutas y relativas. Resultados: Predominaron los pacientes del sexo masculino, con una edad media de 37,4 años. Entre los pacientes, 38 (82,6 por ciento) arribaron del continente africano, la mayoría de ellos de Angola (26,1 por ciento del total de casos). Fue significativa la relación existente entre el supuesto estado no inmune de los pacientes con la severidad del cuadro clínico y presencia de comorbilidades; así como la severidad del cuadro clínico con mayor parasitemia y la especie Plasmodium falciparum. La respuesta al tratamiento resultó excelente con los esquemas combinados utilizados a base de quinina y cloroquina según la especie. Conclusiones: La demora desde el arribo al ingreso hospitalario de los pacientes constituye un riesgo extraordinario para la reintroducción del paludismo en Cuba y para la vida de estos(AU)

Introduction: Malaria is an acute potentially fatal febrile disease caused by parasites transmitted to humans through the bite of mosquitoes from the genus Anopheles. Cuba succeeded in eliminating transmission of this disease thanks to great efforts geared to such an end. It is therefore necessary to take a number of measures aimed at preventing its re-emergence via surveillance and the International Health Control Program. Objective: Clinically characterize a group of patients with imported malaria. Methods: A descriptive cross-sectional study was conducted of 46 adult patients with imported malaria admitted to Pedro Kourí Tropical Medicine Institute from January 2015 to December 2016. The data were collected from the patients' medical records. Results of the analysis of qualitative variables were transferred onto absolute and relative frequency tables. Results: Male patients prevailed, with a mean age of 37.4 years. Of the patients studied, 38 (82.6 percent) were from the African continent, most of them from Angola (26.1 percent of the total cases). A significant relationship was found between the supposed non-immune status of patients and the severity of the clinical status and the presence of comorbidities, as well as between the severity of the clinical status and greater parasitemia and the presence of the species Plasmodium falciparum. An excellent response was obtained to treatment with combined schemes based on quinine and chloroquine, depending on the species. Conclusions: Delay between arrival and hospital admittance of patients is an extraordinary risk for the reintroduction of malaria in Cuba and to the patients' lives(AU)

Immune characteristics of Plasmodium reinfections in mice following chloroquine cure of primary Plasmodium infections / 中国血吸虫病防治杂志

ObjectiveTo investigate the disease progression and immunoprotective characteristics in mice re-infected with homogeneous/heterogeneous Plasmodium strains following cure of Plasmodium infections with chloroquine at the peak of parasitemia. MethodsC57BL/6 mice were infected with the non-lethal P. yoelii 17XNL strain, and half of mice were given treatment with chloroquine at the peak of parasitemia (9 days post-infection), while the other mice were self-cured naturally. Then, all cured mice were re-infected with the equivalent lethal P. yoelii 17XL or P. berghei ANKA strain 90 days following primary Plasmodium infections. The parasitemia levels during primary infections and reinfections were measured by microscopic examinations of Giemsa-stained thin blood films, and the levels of the IgG antibody in sera and the percentages of memory T cell subsets in spleen cells were detected in mice using ELISA and flow cytometry before and after parasite reinfections, respectively. Results Following primary infections with the P. yoelii 17XNL strain, the serum IgG antibody levels were (5.047 ± 0.924) pg/mL in the selfcured mice and (4.429 ± 0.624) pg/mL in the chloroquine-treated mice, respectively (t = 0.437, P > 0.05), which were both significantly higher than that in the uninfected mice (1.624 pg/mL ± 0.280 pg/mL) (F = 22.522, P < 0.01). There was no significant difference in the serum IgG antibody level among self-cured and chloroquine-treated mice re-infected with the P. yoelii 17XL strain or the P. berghei ANKA strain (F = 0.542, P > 0.05); however, the serum IgG antibody levels were all significantly higher in selfcured and chloroquine-treated mice re-infected with the P. yoelii 17XLstrain[(15.487±1.173)pg/mLand(15.965±1.150)pg/mL] or the P. berghei ANKA strain [(14.644 ± 1.523) pg/mL and (15.185 ± 1.333) pg/mL] relative to primary infections (F = 67.383, P < 0.01). There was no significant difference in the proportion of CD4+ [(34.208 ± 2.106), (32.820 ± 1.930), (34.023 ± 2.289), (35.608 ± 1.779) pg/mL] or CD8+ T memory cells [(17.935 ± 2.092), (18.918 ± 2.823), (17.103 ± 1.627), (17.873 ± 1.425) pg/mL] in self-cured and chloroquine-treated mice with primary infections with the P. yoelii 17XNL strain followed by re-infections with the P. yoelii 17XL strain or the P. berghei ANKA strain (F = 0.944 and 0.390, both P > 0.05); however, the proportions of the CD4+ or CD8+ T memory cells were significantly greater in self-cured and chloroquine-treated mice with primary infections with the P. yoelii 17XNL strain followed by re-infections with the P. yoelii 17XL strain or the P. berghei ANKA strain than in mice with primary infections (F = 50.532 and 21.751, both P < 0.01). Conclusions The cure of murine Plasmodium infections with chloroquine does not affect the production of effective immune protections in mice during parasite re-infections. Following a primary infection, mice show a protection against re-infections with either homogeneous or heterogeneous Plasmodium strains, and a higher-level resistance to re-infections with homogeneous parasite strains is found than with heterogeneous strains.

Hallazgo incidental de microfilarias de Mansonella ozzardi en un paciente pediátrico con linfoma T anaplásico / Incidental finding of Mansonella ozzardi in a paediatric patient with anaplastic lymphoma

Resumen Las filariasis son parasitosis producidas por nemátodos hemáticos de la familia Filariidae, la Mansonella ozzardi, es uno de los agentes etiológicos distribuido ampliamente en el continente americano y en el Caribe. Presentamos el caso de una paciente de 13 años de edad, previamente diagnosticada con linfoma T de célula grande anaplásico. Como parte de la evaluación antes del segundo ciclo B de quimioterapia, se realizó un extendido de sangre periférica en el que se encontró una microfilaría tipificada como Mansonella ozzardi, se dio manejo con una dosis única de ivermectina y se logró resultado negativo en el control a las 24 horas. Actualmente la paciente se encuentra asintomática y sin evidencia de recurrencia de la parasitemia y terminando su tratamiento oncológico.

Abstract Filariasis is caused by nematodes in the blood. Mansonella ozzardi is one of the aetiological agents widely distributed in the Americas and the Caribbean. The case is presented on a paediatric patient previously diagnosed with T-cell anaplastic large cell lymphoma. As part of the evaluation before the second cycle B chemotherapy, a peripheral blood smear was performed, in which were found microfilaria, identified as Mansonella ozzardi. The treatment was a single dose of ivermectin, with a negative result being obtained at 24 hours. The patient is currently asymptomatic and with no evidence of recurrence of the parasitaemia, and able to finish the cancer treatment.

Benznidazole therapy for Chagas disease in asymptomatic Trypanosoma cruzi -seropositive former blood donors: evaluation of the efficacy of different treatment regimens

Abstract: INTRODUCTION: Chagas disease currently affects 5.7 million people in Latin America and is emerging in non-endemic countries. There is no consensus concerning the efficacy of trypanocidal therapy for patients with the chronic form of the disease. We evaluated cardiac function and sociodemographic, clinical, and serologic characteristics of a group of asymptomatic Trypanosoma cruzi-seropositive former blood donors, and compared the effects of benznidazole treatment applied for different lengths of time. METHODS: Blood donors who screened positive for T. cruzi between 1998 and 2002 were recruited 10 years later for follow-up (n = 244); 46 individuals had received treatment. Three subjects had terminated treatment prematurely. The remaining 43 individuals were divided into two groups: individuals who had received benznidazole therapy for 50-60 days (n = 28; BT ≤60 group) or more than 60 days (n = 15; BT >60). Serologic assays, biochemical tests, electrocardiographic, echocardiographic, and clinical examinations were performed on all participants. Parasite loads were determined by qualitative and quantitative polymerase chain reaction. RESULTS: Parasitemia was significantly reduced in the BT ≤60 and BT >60 groups compared with the untreated group. There were no differences in epidemiologic profiles or clinical, biochemical, electrocardiographic, or echocardiographic data between any of the groups. CONCLUSIONS: Despite elimination or significant reduction in parasitemia in patients with chronic Chagas disease who received benznidazole, there was no clinical difference between those who were treated for >60 days and those treated for a shorter duration. Furthermore, the adverse effects of benznidazole appear to be less severe than previous reports would suggest.

Indices of Cardiovascular Diseases and Malaria Parisitemia amongst Solid Waste Disposal Workers in Port Harcourt Nigeria.

The aim of this study was to evaluate the health risks of solid waste disposal workers and scavengers, involved in the handling and disposal of municipal wastes, especially their cardiovascular health. The study was carried out in Port Harcourt, Nigeria from 2011 to 2013. Male solid waste disposal workers and scavengers (n=100) aged between 19 and 53years who have worked from six months to 15 years in waste disposal industry, were used in this study. Twenty six percent were aged 40 years and above while 74% were aged between 19 to 39 years. These were compared with 100 male subjects of same age group (21 to 50) who were not solid waste disposal workers; they served as the control group. Their blood pressures (systolic and diastolic) were measured using a standard mercury sphygmomanometer, weight was measured using calibrated electronic scale and height was measured using a Stadiometer. Body Mass Index (BMI) was calculated by the formula (weight/height2). These were used as indices of cardiovascular disease in the volunteers. The level of malaria parasitemia was evaluated. A well-structured questionnaire was given to each volunteer in this study as a behavioral determinant. Mean systolic pressure of the solid waste disposal workers and the control group were 136±13.58 mmHg (Mean ± SD) and 117±12-04 mmHg respectively. The diastolic pressures were 88±12.73 mmHg and 71±10.71 mmHg respectively. This was significant (p<0.01) despite the fact that the solid waste disposal workers had a lower BMI value (24.25±3.19 Kg/M2) than the controls (29.46±3.75 Kg/M2). The increase in blood pressure cuts across duration of exposure, 55.84% of the solid waste disposal workers reported regular alcohol intake, while 43.27% reported current tobacco use against 20% and 7% of the control group. There was no significant difference (p>0.01) when the blood pressures of solid waste disposal workers aged 40 and above was compared with that of 19 to 39 age group. Prevalence of malaria parasitaemia was higher in the solid waste disposal workers in comparison to the controls 92% and 69% respectively.

Hemozoin Pigment: An Important Tool for Low Parasitemic Malarial Diagnosis

Low parasitemic condition in malaria remains a diagnostic challenge; as the available diagnostic methods failed to detect. Currently, hemozoin (Hz) pigment is gaining attention in the diagnosis of malaria. The major drawback is ease of detection of Hz in routine practice. A pilot study was conducted to evaluate the role of Hz pigment and to compare the performance of quantitative buffy coat assay (QBC) and PCR in such conditions. Clinically suspected cases of malaria were examined by both Giemsa stain and immunochromatographic test (ICT). Samples positive by ICT and negative by Giemsa stain were further examined by nested PCR targeting 18S rRNA and QBC for the presence of malaria parasites and pigments. Thirty blood samples fulfilled the inclusion criteria out of which 23 were Plasmodium vivax (Pv), 4 Plasmodium falciparum (Pf), and 3 mixed (Pv and Pf) by immunochromatographic test. Twenty-one out of 30 (70%) were positive by nested PCR in comparison to 25/30 (83%) by QBC. Samples containing both malaria parasites and Hz pigment by QBC completely showed concordance with the PCR result. However, 61% of total samples containing only Hz pigment were observed positive by PCR. Hz pigment remains an important tool for malaria diagnosis. Identification of leukocytes containing pigments by QBC not only indicates recent malarial infections but also puts light on severity of the disease. QBC assay is a rapid, highly sensitive, and cost-effective method to detect malaria parasites and Hz pigment especially in low parasitemic conditions.

Effects of vitaiin c supplementation on the chronic phase of chagas iisease / Efeitos da suplementação de vitamina C na fase crônica da doença de Chagas

Introduction: In order to examine the effectiveness of vitamin C (ascorbic acid) in combating the oxidative insult caused by Trypanosoma cruzi during the development of the chronic phase of Chagas disease, Swiss mice were infected intraperitoneally with 5.0 × 104 trypomastigotes of T. cruzi QM1strain. Methods: Mice were given supplements of two different doses of vitamin C for 180 days. Levels of lipid oxidation (as indicated by thiobarbituric acid reactive substances-TBARS), total peroxide, vitamin C, and reduced glutathione were measured in the plasma, TBARS, total peroxide and vitamin C were measured in the myocardium and histopathologic analysis was undertaken in heart, colon and skeletal muscle. Results: Animals that received a dose equivalent to 500 mg of vitamin C daily showed increased production of ROS in plasma and myocardium and a greater degree of inflammation and necrosis in skeletal muscles than those that received a lower dose or no vitamin C whatsoever. Conclusion: Although some research has shown the antioxidant effect of vitamin C, the results showed that animals subject to a 500 mg dose of vitamin C showed greater tissue damage in the chronic phase of Chagas disease, probably due to the paradoxical actions of the substance, which in this pathology, will have acted as a pro-oxidant or pro-inflammatory. .

Introdução: Para verificar a eficácia da vitamina C em combater o insulto oxidativo causado pelo Trypanosoma cruzi durante a evolução da fase crônica da doença de Chagas, camundongos Swiss foram previamente infectados via intraperitoneal com 5.0 × 104 tripomastigotas da cepa QM1 de T. cruzi. Métodos: Camundongos foram suplementados com duas diferentes doses de vitamina C por 180 dias. Foram mensurados os níveis de peroxidação lipídica (indicado por substâncias reativas ao ácido tiobarbitúrico-TBARS), peróxido total, vitamina C, e glutationa reduzida no plasma e TBARS, peróxido total e vitamina C no miocárdio, e foi realizado o estudo histopatológico em coração, cólon e músculo esquelético. Resultados: Animais que receberam diariamente uma dosagem equivalente a 500 mg de vitamina C apresentaram aumento na produção de ROS e RNS no plasma e no miocárdio e maior grau de inflamação e necrose em músculo esquelético em comparação àqueles que receberam doses menores ou nenhuma vitamina C. Conclusão: Embora muitas pesquisas tenham mostrado o efeito antioxidante da vitamina C, nossos resultados mostraram que os animais que foram expostos a 500 mg de vitamina C apresentaram maior dano tecidual na fase crônica da doença de Chagas, provavelmente devido a ações paradoxais desta substância, onde nesta patologia, poderá agir como pró-oxidante ou pró-inflamatória. .

In vivo antiplasmodial and in vitro antioxidant properties of stem bark extracts of Haematostaphis barteri

Objective: To evaluate the antimalarial and antioxidant properties of stem bark extracts of Haematostaphis barteri (H. barteri). Methods: The prophylactic activity of the plant was performed by dosing mice with sulfadoxine-pyrimethamine (1.2 mg/kg), aqueous extract (30, 100, 300 mg/kg) and dichloromethane/methanol (D/M) (30, 100, 300 mg/kg) extracts of H. barteri for 3 days. On the 4th day, the mice were inoculated with Plasmodium berghei. The parasite density was estimated for each mouse 72 h post-parasite inoculation. The curative activity of the plant was also performed by inoculating mice with Plasmodium berghei. Three days later, they were treated with artemether-lumefantrine (4 mg/kg), aqueous and D/M extracts of H. barteri stembark for 5 days. The in vitro antioxidant property of the aqueous extractwas determined by using the reducing power, nitric oxide and total antioxidant capacity assays. Results: The aqueous extract exerted significant (P < 0.05) curative and prophylactic antimalarial activities. The D/M extract exhibited significant curative (P < 0.05) but not prophylactic antiplasmodial effect. The aqueous extract exhibited in vitro antioxidant property with IC

Parasitemia Induces High Plasma Levels of Interleukin-17 (IL-17) and Low Levels of Interleukin-10 (IL-10) and Transforming Growth Factor-ß (TGF-ß) in Pregnant Mice Infected with Malaria

Background: During pregnancy, the balanced dominance of the T helper17 response shifts to a Th2 response that is characterised by the production of IL-10, following the completion of the implantation process. Transforming growth factor-β (TGF-β) expression is associated with the completion of trophoblast invasion and placental growth. This study assessed the effect of malaria infection on the levels of IL-17, IL-10, and TGF-β in the plasma of pregnant mice with malaria. Methods: Seventeen pregnant BALB/C mice were divided into two groups: mice infected with Plasmodium berghei (treatment group) and uninfected mice (control group). The mice were sacrificed on day 18 post-mating. Parasitemia was measured by Giemsa staining. The levels of IL-17, IL-10, and TGF-β were measured by ELISA. Results: Using independent t test, the IL-17 levels in the treatment group were higher than those in the control group (P = 0.040). The IL-10 levels in the treatment group were lower than those in the control group (P = 0.00). There was no significant difference in the TGF-β levels (P = 0.055) between two groups. However, using SEM analysis the degree of parasitemia decreased the plasma TGF-β levels (tcount = 5.148; ≥ ttable = 1.96). SEM analysis showed that a high degree of parasitemia increased the IL-17 levels and decreased the IL-10 and TGF-β levels. Conclusion: Malaria infection during pregnancy interferes with the systemic balance by increasing the IL-17 levels and decreasing the IL-10 and TGF-β levels.

The Association of Ponderal Benznidazole with its Ultra-high Diluted Formula Reduces the Toxic Effects and Allows Increasing of Dose in Dose-dependent Protocol in Mice Infected with Trypanosoma cruzi

Although several diseases are treated by toxic drugs, their side effects may hamper adherence to the therapy. The aim of this study is to evaluate the effect of the association of ponderal benznidazole (BZ) with its ultra-high diluted (UHD) formula on clinical and parasitological parameters of mice infected by Trypanosoma cruzi (T. cruzi). 24 non-isogenic Swiss mice were divided into groups: CI – infected animals treated with 7% alcohol; BZp – infected animals treated with BZ (500 mg/ kg) from the beginning of infection; BZp+d – infected animals treated with ponderal BZ and with UHD BZ, which started to be administered four days after the beginning of treatment with ponderal BZ; CNI - group of non-treated and non-infected animals. The UHD medicine was prepared according to Phamacopoeia until 30x. The different treatment schedules were statistically compared through parasitological and clinical parameters. The group BZp+d displayed more favorable clinical evolution than the group BZp, with improvement of mass gain, feed conversion and water intake, presenting data approximated to CNI group. The significant increase of the body temperature of BZp+d group indicates an activation of the immune system which was not observed in the other groups. Moreover, the anti-parasitic effect of the ponderal drug was maintained in all parasitological parameters of this group. By reducing the side effects and maintaining the action of the ponderal drug, the combination of toxic drugs with their UHD formula could be considered a way of improving efficacy of the treatment...

A infecção por Trypanossoma cruzi é um problema de saúde pública e o único medicamento disponível no Brasil é o benznidazol (BZ), com efeitos limitados e tóxicos. Estudos anteriores com BZ na dose de 200 mg/kg indicaram que a administração de BZ diluído (30d) controla os efeitos tóxicos da droga em dose ponderal, sem alterar a sua ação terapêutica. Sob essa perspectiva e considerando a ação do BZ dose dependente, aumentar a quantidade de droga administrada significaria uma melhora na eficácia do tratamento. Portanto, este trabalho teve como objetivo avaliar o efeito do BZ ponderal (BZP), na dose de 500 mg/kg associado com BZ diluído (BZD) nos parâmetros clínicos de camundongos infectados por T. cruzi. Em estudo cego, controlado e randomizado, foram utilizados 23 camundongos suíços, machos, com 8 semanas divididos em grupos: CNI - Não infectados e não tratados; CI - Infectados e tratados com álcool 7 %; BZP - Infectados tratados com BZ (500 mg/kg de peso/ animal) a partir do início da infecção; BZP + BZD - Infectados e tratados com a associação de BZP e BZD. Os medicamentos foram administrados por gavagem (0,2 mL/ dia/ animal). O BZP foi administrado a partir da constatação da infecção. O BZ diluído foi preparado de acordo com a Farmacopeia Homeopática Brasileira e administrado 4 dias após o início do tratamento com BZP. Os parâmetros clínicos, avaliados diariamente, incluíram: peso, consumo de ração e água, temperatura e quantidade de excretas. A análise clínica apontou melhores resultados nos grupos BZP e BZP + BZD, mostrando melhor evolução de peso, consumo de ração, água e excretas quando comparado aos grupos não tratados (p< 0.05)...

Analysis of pulmonary manifestations in patient infected with plasmodium vivax / Análise das manifestações pulmonares em pacientes infectados por plasmodium vivax

Objective: To evaluate pulmonary manifestations in patients infected with Plasmodium vivax. MethOds: This was a cross- sectional, retrospective study of 50 patients diagnosed in 2006 to 2008 with vivax malaria at the Evandro Chagas Institute and referred to the University Hospital João de Barros Barreto to examine the pulmonary manifestations. Results: 72% of the patients were men, 28% were 21 to 30 years of age, 30% had parasitaemia 50 to 2000 p/mm3, 88%, 94% and 92% of the patients presented respectively with fever, chills and headache respectively, 56% of the patients had cough, 62% felt breathlessness, 28% presented dyspnea and 86% experienced chest pain. COnClusiOn: The majority of patients surveyed had parasitaemia in the range 50 to 2000 p/mm3. The classic triad fever, chills and headache was present in most patients. Among pulmonary manifestations, cough, chest pain and shortness of breath were reported by the majority of patients.(AU)

Objetivo: Analisar as manifestações pulmonares em pacien- tes infectados por Plasmodium vivax. Métodos: Estudo transversal realizado por meio da análise de 50 prontuários de pacientes diagnosticados com malária vivax entre 2006 a 2008, no Instituto Evandro Chagas, e encaminhados para o Hospital Universitário João de Barros Barreto para análise das manifes- tações pulmonares. ResultadOs: Observou-se que 72% dos pacientes eram homens, 28% possuíam de 21 a 30 anos de idade, 30% apresentaram parasitemia de 50 a 2000 p/mm3. Entre as manifestações clínicas, 88%, 94% e 92% dos pacientes apresentaram, respectivamente, febre, calafrio e cefaleia, 56% apresentaram tosse, 62% sentiram falta de ar, 28% dispneia e 86% dor torácica. COnClusãO: A maioria dos pacientes pesquisados apresentou parasitemia no intervalo de 50 a 2000 p/mm3. A tríade clássica da malária esteve presente na maioria, já dentre as manifestações pulmonares pesquisadas, a tosse, a dor torácica e a falta de ar corresponderam aos sintomas mais relatados pelos pacientes.(AU)

Síndrome de dificultad respiratoria aguda del adulto por Plasmodium vivax / Adult acute respiratory distress syndrome by Plasmodium vivax

El síndrome de dificultad respiratoria aguda del adulto en pacientes con malaria está asociado a infección por Plasmodium falciparum, ocasionalmente manifestado en pacientes infectados por Plasmodium vivax, por lo que han sido pocos los casos reportados en la literatura (1). Reportamos el caso de un paciente de 43 años quien estuvo en área endémica y desarrolló síndrome de dificultad respiratoria aguda del adulto (SDRA) por Plasmodium vivax. El diagnóstico fue realizado por métodos microscópicos. Concluimos que el SDRA asociado a Plasmodium vivax puededesarrollarse antes de iniciar terapia antimalárica, condición con una alta morbimortalidad. (Acta Med Colomb 2014; 39: 211-215).

The adult acute respiratory distress syndrome in patients with malaria is associated with Plasmodium falciparum infection, and only occasionally manifested in patients infected with Plasmodium vivax, so few cases have been reported in the literature. 1 The case of a 43 year old patient who was in an endemic area and developed acute adult respiratory distress syndrome (ARDS) by Plasmodium vivax is reported. The diagnosis was made by microscopic methods. It was concluded that ARDS associated with Plasmodium vivax can develop before starting antimalarial therapy, a condition with high morbidity and mortality. (Acta Med Colomb 2014; 39: 211-215).

Human malaria in C57BL/6J mice: An in vivo model for chemotherapy studies.

The present work deals with the development of Plasmodium falciparum stages in mouse model and its potential for the study of efficacy of antimalarial drugs. C57BL/6J mice were infected with multidrug resistant P. falciparum strain then treated with arteether and artesunate. A response was observed to antimalarial drugs in terms of decrease in parasitemia. Mice infected with P. falciparum strain were successfully cured after treatment with either arteether or artesunate. The speed of parasite clearance time and burden of parasitemia differed for each drug and matched the previously reported observations, hence stressing the relevance of the model. These findings thus suggest that P. falciparum. infected human RBC (iRBC) – C57BL/6J mice can provide a valuable in vivo system and should be included in the short list of animals that can be used for the evaluation of P. falciparum responses to drugs.

Perfil de expressão de microRNAS no miocárdio na infecção aguda pelo Trypanosoma cruzi em camundongos / Expression profile of microRNAs in myocardium during acute infection with Trypanosoma cruzi in mice

A doença de Chagas é uma doença crônica causada pela infecção pelo protozoário Trypanosoma cruzi (T.cruzi). A sua principal consequência clínica é o desenvolvimento da cardiomiopatia chagásica crônica (CCC), que acomete 30% dos pacientes. Não foi determinado um indicador de evolução para a CCC ou permanência na forma indeterminada assintomática da doença de Chagas. Diversos trabalhos têm mostrado alterações no perfil de expressão gênica e proteômica ocorridas na fase aguda e crônica da doença de Chagas experimental e humana. Tais alterações advêm da regulação estabelecida em diversos estágios da expressão gênica e podem ser fatores relevantes no prognóstico da doença. Neste contexto, os microRNAs (miRs), podem exercer uma importante função reguladora. Sua ação se dá pela associação a um RNA mensageiro (RNAm) alvo, inibindo sua tradução ou degradando este transcrito. Assim, a hipótese deste trabalho é a de que a infecção aguda por T. cruzi modula a expressão de miRs no miocárdio de camundongos. Foi avaliado por qRT-PCR o perfil de expressão de miRs 15, 30 e 45 dias após a infecção. O perfil de expressão de miRs resultante foi suficiente para segregar os grupos de acordo com o tempo da infecção. O número de miRs diferencialmente expressos aumentou com a progressão da infecção. Além disso, seis miRs tiveram sua expressão correlacionada à piora na parasitemia e intervalo QTc dos animais: miR-142-3p miR-142-5p, miR-145, miR-146b, miR-149 e miR-21. Análises de correlação realizadas com todos os miRs avaliados ressaltaram este mesmo grupo de miRs entre os mais significativamente correlacionados, além de outros 73 correlacionados com a parasitemia, 67 com o intervalo QTc e 16 com ambos os parâmetros simultaneamente. Nas análises in silico, TNF-alfa e ciclina-D1 foram moléculas nodais recorrentes nas redes criadas com alvos dos miRs diferencialmente expressos em todos os tempos avaliados. Na única rede criada com os miRs correlacionados às alterações...

Chagas disease is a chronic illness caused by infection with the protozoan Trypanosoma cruzi (T. cruzi). Its main clinical outcome is the development of chronic Chagas cardiomyopathy (CCC), which affects 30% of the patients. The factors that define the progression to CCC or maintenance in the asymptomatic indeterminate form of the disease are still poorly understood. Several studies have presented changes occurred in the gene and proteomic expression profiles in both acute and chronic phases of experimental and human Chagas disease. Such changes result from regulation established at different stages of gene expression and may be relevant for the disease prognosis. In this context, microRNAs (miRs) may play an important regulatory function. miRs act by association to a target messenger RNA (mRNA), inhibiting translation or degrading the transcript. Thus, our hypothesis is that acute infection by T. cruzi modulates the expression of microRNAs in the myocardium of mice. The miR expression profile was evaluated by qRT-PCR 15, 30 or 45 days after the infection. This profile was sufficient to segregate the samples according to the time of infection. The number of differentially expressed miRs was higher as the infection progressed. Moreover, six miRs had their expression correlated with worsening of parasitaemia and QTc interval: miR-142-3p miR-142- 5p, miR-145, miR-146b, miR-149 and miR-21. Secondary unbiased correlation analyses showed this cluster of miRs among the most significant and other 73 miRs correlated with parasitaemia, 67 with QTc and 16 with both parameters simultaneously. In silico target prediction analyses showed TNF-alfa and cyclin-D1 as recurrent nodal molecules of the networks created with miRs targets from all time points. The network generated with miRs correlated to changes in parasitaemia and QTc interval showed TNF-alfa, TGF-beta, Rac1 and Src as nodal molecules. This work points out for the first time the involvement of miRs in the acute...

Effects of vitamin C supplementation on acute phase Chagas disease in experimentally infected mice with Trypanosoma cruzi QM1 strain / Efeitos da suplementação de vitamina C sobre a fase aguda da doença de Chagas em camundongos experimentalmente infectados pela cepa QM1 de Trypanosoma cruzi

The tissue changes that occur in Chagas disease are related to the degree of oxidative stress and antioxidant capacity of affected tissue. Studies with vitamin C supplementation did not develop oxidative damage caused by Chagas disease in the host, but other studies cite the use of peroxiredoxins ascorbate - dependent on T. cruzi to offer protection against immune reaction. Based on these propositions, thirty "Swiss" mice were infected with T. cruzi QM1 strain and treated with two different vitamin C doses in order to study the parasitemia evolution, histopathological changes and lipid peroxidation biomarkers during the acute phase of Chagas disease. The results showed that the parasite clearance was greater in animals fed with vitamin C overdose. There were no significant differences regarding the biomarkers of lipid peroxidation and inflammatory process or the increase of myocardium in animals treated with the recommended dosage. The largest amount of parasite growth towards the end of the acute phase suggests the benefit of high doses of vitamin C for trypomastigotes. The supplementation doesn't influence the production of free radicals or the number of amastigote nests in the acute phase of Chagas disease.

As alterações teciduais que ocorrem na doença de Chagas estão relacionadas ao grau de estresse oxidativo e à capacidade antioxidante do tecido afetado. Estudos realizados com suplementação de vitamina C revelaram redução no dano oxidativo causado no hospedeiro pela doença de Chagas, porém outros estudos citam o uso de peroxiredoxinas dependentes de ascorbato pelo T. cruzi para se proteger da ação imune. Com base nessas proposições, trinta camundongos "Swiss" foram infectados com a cepa QM1 de T. cruzi e tratados com duas diferentes doses de vitamina C para estudar a evolução da parasitemia, alterações histopatológicas e dosagem de biomarcadores de peroxidação lipídica durante a fase aguda da doença de Chagas. Os resultados mostraram que a parasitemia foi maior nos animais que receberam uma superdosagem de vitamina C. Não houve diferenças significativas quanto aos biomarcadores de peroxidação lipídica e houve maior processo inflamatório no miocárdio dos animais tratados com dosagem recomendada. O maior crescimento parasitário ao fim da fase aguda sugere benefício de altas doses de vitamina C aos tripomastigotas. A suplementação não exerceu influência sobre a produção de radicais livres e o número de ninhos de amastigotas na fase aguda da doença de Chagas.

Influencia del sexo en la susceptibilidad de ratones Swiss a Trypanosoma cruzi / Sex influence on the susceptibility of Swiss mice to Trypanosoma cruzi

Algunos estudios han reportado que los ratones hembras son más resistentes al trypanosoma cruzi que los machos. Para probar lo anterior, se realizo un estudio comparativo con cuatro cepas de T. cruzi de Costa Rica midiendo parámetros como niveles de parasitemia, porcentajes de mortalidad e histopatología, en un total de 240 ratones (160 para parasitemia y mortalidad y 80 para histopatología). Todas las cepas correspondieron al genotipo TcI. En todos los casos los ratones machos resultaron más susceptibles a la infección (P<0,001). Se observaron parasitemias hasta cinco veces más altas en los machos que, en general, mueren antes que las hembras. La cepa Oswaldo fue la que presentó una mayor mortalidad y niveles de parasitemia más altos. El tejido cardiaco fue el más afectado tanto en los ratones machos como en las hembras, con las cepas bolita y capitán el número de nidos en el miocardio fue significativamente mayor en los machos que en las hembras. Una vez concluido el análisis comparativo se eligió la cepa humana GA, para determinar su efecto en ratones machos y hembras castrados, al lado de ratones normales (150 ratones en total). Las hembras normales se mostraron como las más resistentes, con parasitemias menores que las castradas (P<0,001), y los machos normales como los más susceptibles, con parasitemias significativamente más altas (P<0,001) y supervivencias menores (P<0.01), que los ratones castrados. No se observó diferencia significativa en cuanto al número de nidos en los tejidos entre ratones normales y castrados. Se concluye que el sexo tiene influencia en la resistencia a la infección experimental por T. cruzi.

Some authors have reported that female mice are more resistant to Trypanosoma cruzi infections than male mice. In order to test this hypothesis we did a comparative study using four isolates of T. cruzi from Costa Rica in a total of 160 mice, and observed several parameters such as: parasitemia levels, percent mortality and histopathology. All isolates were identified as genotype TcI. Male mice were more susceptible to infection (P<0.001). Parasitemias reached levels up to five times higher in males and shorter survival periods were observed in males than in females. The Oswaldo isolate showed the highest parasitemias and mortality rates. The heart tissue was the most affected in both males and females. In two isolates the number of parasites in the heart was significantly higher in males than in females. In separate experiments, the human GA isolate was selected in order to observe its effect in castrated male and female mice. Non-castrated females were the most resistant, with lower parasitemias than castrated females, and non-castrated males were the most susceptible with higher parasitemias (P<0.001) and lower survival periods than castrated males (P<0.01). There was no significant difference in the number of parasites in tissues between normal and castrated mice. The final conclusion is that there is a marked sex influence in the susceptibility of mice to T. cruzi experimental infections.