Development of chimeric protein as a multivalent vaccine for human Kinetoplastid infections: Chagas disease and leishmaniasis.

Leishmania spp. and Trypanosoma cruzi are parasitic kinetoplastids of great medical and epidemiological importance since they are responsible for thousands of deaths and disability-adjusted life-years annually, especially in low- and middle-income countries. Despite efforts to minimize their impact, current prevention measures have failed to fully control their spread. There are still no vaccines available. Taking into account the genetic similarity within the Class Kinetoplastida, we selected CD8+ T cell epitopes preserved among Leishmania spp. and T. cruzi to construct a multivalent and broad-spectrum chimeric polyprotein vaccine. In addition to inducing specific IgG production, immunization with the vaccine was able to significantly reduce parasite burden in the colon, liver and skin lesions from T. cruzi, L. infantum and L. mexicana challenged mice, respectively. These findings were supported by histopathological analysis, which revealed decreased inflammation in the colon, a reduced number of degenerated hepatocytes and an increased proliferation of connective tissue in the skin lesions of the corresponding T. cruzi, L. infantum and L. mexicana vaccinated and challenged mice. Collectively, our results support the protective effect of a polyprotein vaccine approach and further studies will elucidate the immune profile associated with this protection. Noteworthy, our results act as conceptual proof that a single multi-kinetoplastida vaccine can be used effectively to control different infectious etiologies, which in turn can have a profound impact on the development of a new generation of vaccines.

Nitric oxide contributes to liver inflammation and parasitic burden control in Ascaris suum infection.

BACKGROUND: Human ascariasis is one of the most prevalent neglected tropical diseases worldwide. The immune response during human ascariasis is characterized by Th2 polarization and a mixed Th2/Th17 response during the pathogenesis of experimental larval ascariasis. Cytokines and other pro-inflammatory mediators, such as nitric oxide (NO), are involved in helminthic infections. However, the role of NO in ascariasis remains unclear. OBJECTIVES: Given the importance of NO in inflammation, we aimed to determine the immunological and histopathological alterations in the livers of C57BL/6 iNOS-/- mice during A. suum infection. METHODS: In this study, parasitic load was evaluated in the livers of wild type C57BL/6 and C57BL/6 iNOS-/- mice infected with A. suum. Histopathological and morphometric analyses and analysis of serum cytokines via Cytometric Bead Array were performed, and the activity of eosinophil peroxidase and myeloperoxidase of neutrophils in the tissues were determined. RESULTS: The results showed that NO is important for controlling parasitic load during infection by A. suum. C57BL/6iNOS-/- mice showed reduced inflammatory processes and less tissue damage during liver larval migration of A. suum, which is associated with a reduction in serum levels of pro-inflammatory cytokines. CONCLUSIONS: We demonstrated that NO is a crucial inflammatory molecule during Ascaris sp. infection and controls the establishment of the parasite and the development of the host immune response in the liver.

Genetic Background Affects the Mucosal Secretory IgA Levels, Parasite Burden, Lung Inflammation, and Mouse Susceptibility to Ascaris suum Infection.

Ascariasis is a neglected tropical disease that is widespread in the world and has important socioeconomic impacts. The presence of various stages of worm development in the pulmonary and intestinal mucosae induces a humoral and cellular immune response. However, although there is much evidence of the protective role of mucosal immunity against various pathogens, including helminths, there is still a gap in the knowledge about the immune response and the mechanisms of action that are involved in protection against diseases, especially in the initial phase of ascariasis. Thus, the aim of this study was to evaluate the kinetic aspects of the immune parasitological parameters in intestinal and pulmonary mucosae in male mice with early ascariasis. Therefore, two mouse strains that showed different susceptibilities to ascariasis (BALB/c and C57BL/6J) when experimentally infected with 2,500 infective eggs of Ascaris suum from time point 0 were examined: the immune parasitological parameters were evaluated each 2 days after infection over a period of 12 days. The results were suggestive of a synergetic action of intestinal and pulmonary secretory IgA (S-IgA) contributing to protection against early ascariasis by reducing the amount of migrating larvae as well as the influx of leukocytes in the lung and the consequent impairment of pulmonary capacity.

Eosinophils mediate SIgA production triggered by TLR2 and TLR4 to control Ascaris suum infection in mice.

Human ascariasis is the most prevalent but neglected tropical disease in the world, affecting approximately 450 million people. The initial phase of Ascaris infection is marked by larval migration from the host's organs, causing mechanical injuries followed by an intense local inflammatory response, which is characterized mainly by neutrophil and eosinophil infiltration, especially in the lungs. During the pulmonary phase, the lesions induced by larval migration and excessive immune responses contribute to tissue remodeling marked by fibrosis and lung dysfunction. In this study, we investigated the relationship between SIgA levels and eosinophils. We found that TLR2 and TLR4 signaling induces eosinophils and promotes SIgA production during Ascaris suum infection. Therefore, control of parasite burden during the pulmonary phase of ascariasis involves eosinophil influx and subsequent promotion of SIgA levels. In addition, we also demonstrate that eosinophils also participate in the process of tissue remodeling after lung injury caused by larval migration, contributing to pulmonary fibrosis and dysfunction in re-infected mice. In conclusion, we postulate that eosinophils play a central role in mediating host innate and humoral immune responses by controlling parasite burden, tissue inflammation, and remodeling during Ascaris suum infection. Furthermore, we suggest that the use of probiotics can induce eosinophilia and SIgA production and contribute to controlling parasite burden and morbidity of helminthic diseases with pulmonary cycles.

ASCVac-1, a Multi-Peptide Chimeric Vaccine, Protects Mice Against Ascaris suum Infection.

Control of human ascariasis, the most prevalent neglected tropical disease globally affecting 450 million people, mostly relies on mass drug administration of anthelmintics. However, chemotherapy alone is not efficient due to the high re-infection rate for people who live in the endemic area. The development of a vaccine that reduces the intensity of infection and maintains lower morbidity should be the primary target for infection control. Previously, our group demonstrated that immunization with crude Ascaris antigens in mice induced an IgG-mediated protective response with significant worm reduction. Here, we aimed to develop a multipeptide chimera vaccine based on conserved B-cell epitopes predicted from 17 common helminth proteomes using a bioinformatics algorithm. More than 480 B-cell epitopes were identified that are conserved in all 17 helminths. The Ascaris-specific epitopes were selected based on their reactivity to the pooled sera of mice immunized with Ascaris crude antigens or infected three times with A. suum infective eggs. The top 35 peptides with the strongest reactivity to Ascaris immune serum were selected to construct a chimeric antigen connected in sequence based on conformation. This chimera, called ASCVac-1, was produced as a soluble recombinant protein in an Escherichia coli expression system and, formulated with MPLA, was used to immunize mice. Mice immunized with ASCVac-1/MPLA showed around 50% reduced larvae production in the lungs after being challenged with A. suum infective eggs, along with significantly reduced inflammation and lung tissue/function damage. The reduced parasite count and pathology in infected lungs were associated with strong Th2 immune responses characterized by the high titers of antigen-specific IgG and its subclasses (IgG1, IgG2a, and IgG3) in the sera and significantly increased IL-4, IL-5, IL-13 levels in lung tissues. The reduced IL-33 titers and stimulated eosinophils were also observed in lung tissues and may also contribute to the ASCVac-1-induced protection. Taken together, the preclinical trial with ASCVac-1 chimera in a mouse model demonstrated its significant vaccine efficacy associated with strong IgG-based Th2 responses, without IgE induction, thus reducing the risk of an allergic response. All results suggest that the multiepitope-based ASCVac-1 chimera is a promising vaccine candidate against Ascaris sp. infections.

ASCVac-1, a multi-peptide chimeric vaccine, protects mice against Ascaris suum infection / ASCVac-1, uma vacina quimérica multipeptídica, protege camundongos contra a infecção por Ascaris suum

Control of human ascariasis, the most prevalent neglected tropical disease globally affecting 450 million people, mostly relies on mass drug administration of anthelmintics. However, chemotherapy alone is not efficient due to the high re-infection rate for people who live in the endemic area. The development of a vaccine that reduces the intensity of infection and maintains lower morbidity should be the primary target for infection control. Previously, our group demonstrated that immunization with crude Ascaris antigens in mice induced an IgG-mediated protective response with significant worm reduction. Here, we aimed to develop a multipeptide chimera vaccine based on conserved B-cell epitopes predicted from 17 common helminth proteomes using a bioinformatics algorithm. More than 480 B-cell epitopes were identified that are conserved in all 17 helminths. The Ascaris-specific epitopes were selected based on their reactivity to the pooled sera of mice immunized with Ascaris crude antigens or infected three times with A. suum infective eggs. The top 35 peptides with the strongest reactivity to Ascaris immune serum were selected to construct a chimeric antigen connected in sequence based on conformation. This chimera, called ASCVac-1, was produced as a soluble recombinant protein in an Escherichia coli expression system and, formulated with MPLA, was used to immunize mice. Mice immunized with ASCVac-1/MPLA showed around 50% reduced larvae production in the lungs after being challenged with A. suum infective eggs, along with significantly reduced inflammation and lung tissue/function damage. The reduced parasite count and pathology in infected lungs were associated with strong Th2 immune responses characterized by the high titers of antigen-specific IgG and its subclasses (IgG1, IgG2a, and IgG3) in the sera and significantly increased IL-4, IL-5, IL-13 levels in lung tissues. The reduced IL-33 titers and stimulated eosinophils were also observed in lung tissues and may also contribute to the ASCVac-1-induced protection. Taken together, the preclinical trial with ASCVac-1 chimera in a mouse model demonstrated its significant vaccine efficacy associated with strong IgG-based Th2 responses, without IgE induction, thus reducing the risk of an allergic response. All results suggest that the multiepitope-based ASCVac-1 chimera is a promising vaccine candidate against Ascaris sp. infections.

Ketamine can be produced by Pochonia chlamydosporia: an old molecule and a new anthelmintic?

BACKGROUND: Infection by nematodes is a problem for human health, livestock, and agriculture, as it causes deficits in host health, increases production costs, and incurs a reduced food supply. The control of these parasites is usually done using anthelmintics, which, in most cases, have not been fully effective. Therefore, the search for new molecules with anthelmintic potential is necessary. METHODS: In the present study, we isolated and characterized molecules from the nematophagous fungus Pochonia chlamydosporia and tested these compounds on three nematodes: Caenorhabditis elegans; Ancylostoma ceylanicum; and Ascaris suum. RESULTS: The ethyl acetate extract showed nematicidal activity on the nematode model C. elegans. We identified the major substance present in two sub-fractions of this extract as ketamine. Then, we tested this compound on C. elegans and the parasites A. ceylanicum and A. suum using hamsters and mice as hosts, respectively. We did not find a difference between the animal groups when considering the number of worms recovered from the intestines of animals treated with ketamine (6 mg) and albendazole (P > 0.05). The parasite burden of larvae recovered from the lungs of mice treated with ketamine was similar to those treated with ivermectin. CONCLUSIONS: The results presented here demonstrate the nematicidal activity of ketamine in vitro and in vivo, thus confirming the nematicidal potential of the molecule present in the fungus P. chlamydosporia may consist of a new method of controlling parasites.

Evaluation of the euthanasia of seropositive dogs for canine visceral leishmaniasis as the only method of controling the disease in the enzootic area in the Midwestern Minas Gerais / Avaliação da eutanásia de cães soro reagentes para leishmaniose visceral canina como único método de controle da doença em área enzoótica no Centro Oeste de Minas Gerais

Human Visceral Leishmaniasis (VL) is expanding, and it is distributed in the five geographic regions of Brazil, with the Northeast being the region that presents the most significant number of registered cases. The main urban reservoir of the etiological agent of VL is the domestic dog, and it is known that canine cases precede human cases. One of the control actions of VL, recommended by the Ministry of Health, is performing the euthanasia of seropositive dogs. In 2013, in the municipality of Iguatama/MG, the first canine serological survey was carried out in the city, with a prevalence of 8.3% of seropositive dogs for Leishmania infantum. Of the seropositive animals for Canine Visceral Leishmaniasis (CVL) in this survey, 84% were euthanized, and 16% died within two years after diagnosis. No other control measures for CVL were taken in the municipality. The objectives of the present study were to determine the current prevalence of seropositive dogs for CVL in the municipality of Iguatama and to observe the impact of euthanasia of seropositive dogs to VL as the only control measure performed in this municipality, which is considered an enzootic area of the disease. For this, a new canine epidemiological survey was carried out in the municipality of Iguatama, following the guidelines of the Manual of Surveillance and Control of Visceral Leishmaniasis. Blood samples were collected from 270 dogs and tested by DPP® immunochromatographic test and by Enzyme-Linked Immunosorbent Assay (ELISA). The animals that presented inconclusive results had new blood samples collected and analyzed. The only animal that showed a positive result in DPP® and undetermined in the ELISA, in the first evaluation, became seropositive four months later, so this study reinforces the recommendation of the Ministry of Health to reassess dogs that present inconclusive results for CVL. From a total of 270 samples, 21 (7.8%) were reagents in both tests. Thus, the prevalence of seropositive dogs for CVL, in 2017, in the urban area of the municipality of Iguatama was 7.8%. The Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) molecular technique confirmed infection by L. infantum in the ten dogs sampled with CVL-positive serology. The comparison between the current prevalence (after euthanasia) with that found in 2013 (8.3%), there was no significant difference (p=0.764). The prevalence of CVL in the urban area of the municipality of Iguatama is still high, making it possible to presume that the euthanasia of seropositive dogs for CVL, and the absence of monitoring or other interventions measures, did not contribute to decrease the transmission of the parasite and was not sufficient for the control of CVL in the municipality of Iguatama, an area considered as non-endemic to CVL until 2013.(AU)

A Leishmaniose visceral humana (LV) está em franca expansão e distribuída nas cinco regiões geográficas do Brasil, o Nordeste sendo a região em que há o maior número de casos registrados. O principal reservatório urbano do agente etiológico da LV é o cão doméstico e sabe-se que casos caninos antecedem o aparecimento de casos humanos. Uma das ações de controle da LV, preconizada pelo Ministério da Saúde, é a eutanásia dos cães soro reagentes. Em 2013, no município de Iguatama/MG, foi realizado o primeiro inquérito sorológico canino na cidade, sendo constatada uma prevalência de 8,3% de cães soro reagentes para Leishmania infantum. Dos animais com sorologia positiva para Leishmaniose visceral canina (LVC), neste inquérito, 84% foram eutanasiados e 16% foram a óbito em, no máximo, dois anos após o diagnóstico. Nenhuma outra medida de controle para LVC foi realizada no município. Os objetivos deste estudo foram determinar a prevalência atual de cães soro reagentes para LVC no município de Iguatama e observar o impacto das eutanásias de cães soro reagentes para LV como única medida de controle realizada neste município, considerado área enzoótica para a doença. Para isso, foi realizado um novo inquérito epidemiológico canino no município de Iguatama, seguindo as normas do Manual de Vigilância e Controle da Leishmaniose Visceral. Foram colhidas amostras sanguíneas de 270 cães que foram examinadas pelo teste imunocromatográfico DPP® e pelo ensaio imunoenzimático (ELISA). Os animais que apresentaram resultados inconclusivos tiveram novas amostras de sangue coletadas e analisadas. O único animal que apresentou resultado positivo no DPP® e indeterminado no ELISA, na primeira avaliação, soro converteu quatro meses depois, o que reforça a recomendação do Ministério da Saúde de reavaliar os cães que apresentam resultado inconclusivo para LVC. Do total das 270 amostras, 21 foram reagentes nos dois testes. Portanto, a prevalência de cães soro reagentes para LVC, em 2017, na área urbana do município de Iguatama foi igual a 7,8%. A infecção pela espécie Leishmania infantum foi confirmada através da técnica molecular de PCR-RFLP em 10 cães amostrados com sorologia positiva para LVC. A comparação entre a prevalência atual (depois das eutanásias) com a encontrada em 2013 (8,3%), não mostrou diferença significativa (p=0,764). Uma vez que a soro prevalência de LVC na área urbana do município de Iguatama ainda é alta, pode-se supor que a eutanásia dos cães soro reagentes para LVC, somada à ausência de monitoramento e de outras medidas de intervenção, não contribuiu para a diminuição da transmissão do parasito, e não foi suficiente para o controle de LVC no município de Iguatama, uma área considerada, até 2013, indene para LVC.(AU)

Epidemiological profile of soccer-related injuries in a state Brazilian championship: An observational study of 2014-15 season.

BACKGROUND: Soccer related injuries are often reported in studies, but epidemiological research on this theme is rare in Brazil, Furthermore, the conditions in which athletes have returned to sports practice, namely, either symptomatic or asymptomatic, have been neglected in research. OBJECTIVES: The aim of this study was to describe the epidemiological features of injuries among professional Brazilian soccer players in relation to location, type, mechanism, severity, recurrence, treatment and, lastly, symptoms in return to sport. STUDY DESIGN: Descriptive and cross-sectional, observational study. STUDY CENTER: School of Physical Therapy of the University of Mato Grosso do Sul, Brazil. METHODS: 116 male professional athletes of teams from a Brazilian state championship were interviewed and information about injuries was recorded using a retrospective reported morbidity questionnaire. Data were analyzed in mean ±â€¯SD for physical characteristics and sports practice history in absolute and relative frequencies (chi-square test with Bonferroni's correction) for characterization of soccer injuries in terms of type, location, severity, recurrence and symptoms in return to sport. RESULTS: The numbers of injuries per athlete and per injured athlete were 0.92 and 1.43 respectively. The injuries of muscle-tendon unit and the joint types localized on lower limbs constituted the most important clinical occurrences with significant difference both in relation to other types (p < 0.05). Moderate and severe injuries were the most frequent occurrences. In relation to mechanisms for each type of injury, body contact was at least three times more responsible for injury cases. This type of mechanism was associated with a significantly greater impairment of joint structures. Concerning occurrence and recurrence of cases, the number of recurrent injuries of the muscle-tendon unit reached about 7.5% of the first-time injuries, while the number of joint recurrent injuries integrated almost 40% of the first-time cases. Significant differences between first-time injuries and recurrent injuries were found only for muscle-tendon and joint structures (p < 0.05), while significant differences among the type of injuries within each type of occurrence (first-time or recurrent injuries) were also found between muscle-tendon and joint injuries (p < 0.05). In relation to athletes with symptoms, in return to sport, 77.6% of them were treated for their injuries but more than half of them returned with symptoms still present when compared to those who returned without any symptoms. Among athletes who did not receive treatment, a lower percentage (58.3%) returned to the sport with symptoms still present. Significant associations between treatment and symptomatology were not found.

Cardiovascular effects of a strength test (1rm) in prehypertensive subjects / Efeitos cardiovasculares de um teste de força (1rm) em sujeitos pré-hipertensos / Efectos cardiovasculares de un test de fuerza (1rm) en sujetos pre-hipertensos

ABSTRACT Introduction: Physical exercise promotes positive physiological adaptations, reducing the risk of developing cardiovascular diseases and promoting greater autonomic control of heart rate and reduction in pressure levels. Objective: To compare the behavior of arterial pressure and autonomic heart rate modulation in the 1-repetition maximum knee extension test, performed on the 45-degree leg press machine in normotensive and prehypertensive subjects. Methods: Twenty-four male volunteers were assessed. Blood pressure (BP) and heart rate variability (HRV) were evaluated at rest, immediately after the maximum load achieved and during the 10, 20, 30 and 40-minute recovery phases. Statistical analyses were obtained using SigmaStat 3.5 software and a level of significance of 5%. Results: Mean and standard deviation were used to identify 12 normotensive patients (N) aged = 25.5 ± 5.7 years; height = 174.3 ± 6.6 cm; BMI = 24.9 ± 2.4; SBP on average = 112.6 ± 5.6 mmHg and 12 prehypertensive (PH) patients aged = 29.8 ± 6.5 years; height = 175.7 ± 6.7 cm; BMI = 27.8 ± 6.5; mean SBP = 131.1 ± 3.2 mmHg. There was a statistical difference in the pNN50 index (%) at rest (PH = 1.13 ± 0.66 and N = 1.58 ± 0.35) and in the LF/HF index, also at rest (PH = 0.18 ± 0.29 and N = −0.18 ± 0.36). Conclusion: The one-repetition maximum (1RM) test did not promote cardiovascular risk in the prehypertensive group during its execution, with double product results lower than 30,000 mmHg.bpm, demonstrating lower cardiac overload. Moreover, HRV exhibited similarity in sympathetic-vagal behavior, in the time/frequency domain, between the groups. Level of Evidence III; Study Case-control.

RESUMO Introdução: O exercício físico promove adaptações fisiológicas de caráter positivo, diminuindo os riscos de desenvolvimento de doenças cardiovasculares, além de promover um maior controle autonômico da frequência cardíaca e redução nos níveis pressóricos. Objetivo: Comparar o comportamento da pressão arterial e da modulação autonômica da frequência cardíaca no teste de uma repetição máxima de extensão de joelhos, realizado no aparelho de Leg press 45° em sujeitos normotensos e pré-hipertensos. Métodos: Foram avaliados 24 voluntários do sexo masculino. Foi feita a avaliação da pressão arterial (PA) e da variabilidade da frequência cardíaca (VFC) no momento de repouso, imediatamente após a carga máxima alcançada e durante as fases de 10, 20, 30 e 40 minutos de recuperação. As análises estatísticas foram obtidas através do software SigmaStat 3.5 e nível de significância igual a 5%. Resultados: A média e o desvio padrão foram utilizados para identificação de 12 normotensos (N) com idade = 25,5 ± 5,7 anos; altura = 174,3 ± 6,6 cm; IMC = 24,9 ± 2,4; PAS em média = 112,6 ± 5,6 mmHg e 12 pré-hipertensos (PH) com idade = 29,8 ± 6,5 anos; altura = 175,7 ± 6,7 cm; IMC = 27,8 ± 6,5; PAS em média = 131,1 ± 3,2 mmHg. Houve diferença estatística no índice pNN50 (%) no momento repouso (PH = 1,13 ± 0,66 e N =1,58 ± 0,35) e no índice LF/HF igualmente no momento repouso (PH = 0,18 ± 0,29 e N = −0,18 ± 0,36). Conclusão: O teste de uma repetição máxima (1RM) não promoveu risco cardiovascular no grupo pré-hipertenso durante sua execução, com resultados de duplo produto menor que 30.000 mmHg.bpm, demonstrando menor sobrecarga cardíaca, e a VFC apresentou similaridade no comportamento simpático-vagal no domínio do tempo e frequência entre os grupos. Nível de evidência III; Estudo Caso controle.

RESUMEN Introducción: El ejercicio físico promueve adaptaciones fisiológicas de carácter positivo, disminuyendo los riesgos de desarrollo de enfermedades cardiovasculares, además de promover un mayor control autonómico de la frecuencia cardíaca y reducción en los niveles presóricos. Objetivo: Comparar el comportamiento de la presión arterial y de la modulación autonómica de la frecuencia cardíaca en el test de una repetición máxima de extensión de rodillas, realizado en el aparato de Leg press 45° en sujetos normotensos y pre-hipertensos. Métodos: Fueron evaluados 24 voluntarios del sexo masculino. Fue hecha la evaluación de la presión arterial (PA) y de la variabilidad de la frecuencia cardíaca (VFC) en el momento de reposo, inmediatamente después de la carga máxima alcanzada y durante las fases de 10, 20, 30 y 40 minutos de recuperación. Los análisis estadísticos fueron obtenidos a través del software SigmaStat 3.5 y nivel de significancia igual a 5%. Resultados: El promedio y la desviación estándar fueron utilizados para identificación de 12 normotensos (N) con edad = 25,5 ± 5,7 años; altura = 174,3 ± 6,6 cm; IMC = 24,9 ± 2,4; PAS en promedio = 112,6 ± 5,6 mmHg y 12 pre-hipertensos (PH) con edad = 29,8 ± 6,5 años; altura = 175,7 ± 6,7 cm; IMC = 27,8 ± 6,5; PAS en promedio = 131,1 ± 3,2 mmHg. Hubo diferencia estadística en el índice pNN50 (%) en el momento reposo (PH = 1,13 ± 0,66 y N =1,58 ± 0,35) y en el índice LF/HF igualmente en el momento reposo (PH = 0,18 ± 0,29 y N = −0,18 ± 0,36). Conclusión: El test de una repetición máxima (1RM) no promovió riesgo cardiovascular en el grupo pre-hipertenso durante su ejecución, con resultados de doble producto menor que 30.000 mmHg.bpm, demostrando menor sobrecarga cardíaca, y la VFC presentó similitud en el comportamiento simpático-vagal en el dominio del tiempo y frecuencia entre los grupos. Nivel de evidencia III; Estudio Caso control.

IgG Induced by Vaccination With Ascaris suum Extracts Is Protective Against Infection.

Human ascariasis has a global and cosmopolitan distribution, and has been characterized as the most prevalent neglected tropical disease worldwide. The development of a preventive vaccine is highly desirable to complement current measures required for this parasitic infection control and to reduce chronic childhood morbidities. In the present study, we describe the mechanism of protection elicited by a preventive vaccine against ascariasis. Vaccine efficacy was evaluated after immunization with three different Ascaris suum antigen extracts formulated with monophosphoryl lipid A (MPLA) as an adjuvant: crude extract of adult worm (ExAD); crude extract of adult worm cuticle (CUT); and crude extract of infective larvae (L3) (ExL3). Immunogenicity elicited by immunization was assessed by measuring antibody responses, cytokine production, and influx of tissue inflammatory cells. Vaccine efficacy was evaluated by measuring the reductions in the numbers of larvae in the lungs of immunized BALB/c mice that were challenged with A. suum eggs. Moreover, lung physiology and functionality were tested by spirometry to determine clinical efficacy. Finally, the role of host antibody mediated protection was determined by passive transfer of serum from immunized mice. Significant reductions in the total number of migrating larvae were observed in mice immunized with ExL3 61% (p < 0.001), CUT 59% (p < 0.001), and ExAD 51% (p < 0.01) antigens in comparison with non-immunized mice. For the Ascaris antigen-specific IgG antibody levels, a significant and progressive increase was observed with each round of immunization, in association with a marked increase of IgG1 and IgG3 subclasses. Moreover, a significant increase in concentration of IL-5 and IL-10 (pre-challenge) in the blood and IL-10 in the lung tissue (post-challenge) was induced by CUT immunization. Finally, ExL3 and CUT-immunized mice showed a marked improvement in lung pathology and tissue fibrosis as well as reduced pulmonary dysfunction induced by Ascaris challenge, when compared to non-immunized mice. Moreover, the passive transfer of specific IgG antibodies from ExL3, CUT, and ExAD elicited a protective response in naïve mice, with significant reductions in parasite burdens in lungs of 65, 64, and 64%, respectively. Taken together, these studies indicated that IgG antibodies contribute to protective immunity.

Evaluation of diagnostic methods for the detection of intestinal schistosomiasis in endemic areas with low parasite loads: Saline gradient, Helmintex, Kato-Katz and rapid urine test.

BACKGROUND: In some tropical countries, such as Brazil, schistosomiasis control programs have led to a significant reduction in the prevalence and parasite burden of endemic populations. In this setting, the Kato-Katz technique, as the standard diagnostic method for the diagnosis of Schistosoma mansoni infections, which involves the analysis of two slides from one fecal sample, loses its sensitivity. As a result, a significant number of infected individuals are not detected. The objective of this study was to perform extensive parasitological testing of up to three fecal samples and include a rapid urine test (POC-CCA) in a moderate prevalence area in Northern Minas Gerais, Brazil, and evaluate the performance of each test separately and in combination. METHODS AND FINDINGS: A total of 254 individuals were examined with variants of the standard Kato-Katz technique (up to18 Kato-Katz slides prepared from three fecal samples), a modified Helmintex (30 g of feces), the saline gradient (500 mg of feces), and the POC-CCA methods. We established a reference standard taking into consideration all the positive results in any of the parasitological exams. Evaluation of the parasite burden by two Kato-Katz slides confirmed that most of the individuals harbored a light infection. When additional slides and different parasitological methods were included, the estimated prevalence rose 2.3 times, from 20.4% to 45.9%. The best sensitivity was obtained with the Helmintex method (84%). All parasitological methods readily detected a high or moderate intensity of infection; however, all lost their high sensitivity in the case of low or very low intensity infections. The overall sensitivity of POC-CCA (64.9%) was similar to the six Kato-Katz slides from three fecal samples. However, POC-CCA showed low concordance (κ = 0.34) when compared with the reference standard. CONCLUSIONS: The recommended Kato-Katz method largely underestimated the prevalence of S. mansoni infection. Because the best performance was achieved with a modified Helmintex method, this technique might serve as a more precise reference standard. An extended number of Kato-Katz slides in combination with other parasitological methods or with POC-CCA was able to detect more than 80% of egg-positive individuals; however, the rapid urine test (POC-CCA) produced a considerable percentage of false positive results.

IgG induced by vaccination with Ascaris suum extracts is protective against infection

Human ascariasis has a global and cosmopolitan distribution, and has been characterized as the most prevalent neglected tropical disease worldwide. The development of a preventive vaccine is highly desirable to complement current measures required for this parasitic infection control and to reduce chronic childhood morbidities. In the present study, we describe the mechanism of protection elicited by a preventive vaccine against ascariasis. Vaccine efficacy was evaluated after immunization with three different Ascaris suum antigen extracts formulated with monophosphoryl lipid A (MPLA) as an adjuvant: crude extract of adult worm (ExAD); crude extract of adult worm cuticle (CUT); and crude extract of infective larvae (L3) (ExL3). Immunogenicity elicited by immunization was assessed by measuring antibody responses, cytokine production, and influx of tissue inflammatory cells. Vaccine efficacy was evaluated by measuring the reductions in the numbers of larvae in the lungs of immunized BALB/c mice that were challenged with A. suum eggs. Moreover, lung physiology and functionality were tested by spirometry to determine clinical efficacy. Finally, the role of host antibody mediated protection was determined by passive transfer of serum from immunized mice. Significant reductions in the total number of migrating larvae were observed in mice immunized with ExL3 61% (p < 0.001), CUT 59% (p < 0.001), and ExAD 51% (p < 0.01) antigens in comparison with non-immunized mice. For the Ascaris antigen-specific IgG antibody levels, a significant and progressive increase was observed with each round of immunization, in association with a marked increase of IgG1 and IgG3 subclasses. Moreover, a significant increase in concentration of IL-5 and IL-10 (pre-challenge) in the blood and IL-10 in the lung tissue (post-challenge) was induced by CUT immunization. Finally, ExL3 and CUT-immunized mice showed a marked improvement in lung pathology and tissue fibrosis as well as reduced pulmonary dysfunction induced by Ascaris challenge, when compared to non-immunized mice. Moreover, the passive transfer of specific IgG antibodies from ExL3, CUT, and ExAD elicited a protective response in naïve mice, with significant reductions in parasite burdens in lungs of 65, 64, and 64%, respectively. Taken together, these studies indicated that IgG antibodies contribute to protective immunity.

IgG induced by vaccination with Ascaris suum extracts is protective against infection

Human ascariasis has a global and cosmopolitan distribution, and has been characterized as the most prevalent neglected tropical disease worldwide. The development of a preventive vaccine is highly desirable to complement current measures required for this parasitic infection control and to reduce chronic childhood morbidities. In the present study, we describe the mechanism of protection elicited by a preventive vaccine against ascariasis. Vaccine efficacy was evaluated after immunization with three different Ascaris suum antigen extracts formulated with monophosphoryl lipid A (MPLA) as an adjuvant: crude extract of adult worm (ExAD); crude extract of adult worm cuticle (CUT); and crude extract of infective larvae (L3) (ExL3). Immunogenicity elicited by immunization was assessed by measuring antibody responses, cytokine production, and influx of tissue inflammatory cells. Vaccine efficacy was evaluated by measuring the reductions in the numbers of larvae in the lungs of immunized BALB/c mice that were challenged with A. suum eggs. Moreover, lung physiology and functionality were tested by spirometry to determine clinical efficacy. Finally, the role of host antibody mediated protection was determined by passive transfer of serum from immunized mice. Significant reductions in the total number of migrating larvae were observed in mice immunized with ExL3 61% (p < 0.001), CUT 59% (p < 0.001), and ExAD 51% (p < 0.01) antigens in comparison with non-immunized mice. For the Ascaris antigen-specific IgG antibody levels, a significant and progressive increase was observed with each round of immunization, in association with a marked increase of IgG1 and IgG3 subclasses. Moreover, a significant increase in concentration of IL-5 and IL-10 (pre-challenge) in the blood and IL-10 in the lung tissue (post-challenge) was induced by CUT immunization. Finally, ExL3 and CUT-immunized mice showed a marked improvement in lung pathology and tissue fibrosis as well as reduced pulmonary dysfunction induced by Ascaris challenge, when compared to non-immunized mice. Moreover, the passive transfer of specific IgG antibodies from ExL3, CUT, and ExAD elicited a protective response in naïve mice, with significant reductions in parasite burdens in lungs of 65, 64, and 64%, respectively. Taken together, these studies indicated that IgG antibodies contribute to protective immunity.

Could current factors be associated with retrospective sports injuries in Brazilian jiu-jitsu? A cross-sectional study.

BACKGROUND: Brazilian jiu-jitsu is characterized by musculoskeletal disorders and high occurrence of sports injuries. The present study was aimed to analyze some internal factors, as well as to describe occurrence and characteristics of retrospective musculoskeletal injuries in different age groups of Brazilian jiu-jitsu practitioners. METHODS: One hundred ninety-three Brazilian jiu-jitsu practitioners, which were divided into three age groups: Adolescent, Adult, and Master. Besides anthropometric characterization, standard clinical tests were conducted to analyze the global and segmental joint flexibility, lumbar spine range of motion, and handgrip strength. Sports injury occurrence and total physical activity were obtained from an adapted morbidity survey and International Physical Activity Questionnaire - Short Form (IPAQ-SF), respectively. RESULTS: A total of 247 cases of retrospective injuries was registered (1.27 injury/ participant). Occurrence of rectus femoral muscle retraction in the right leg was increased within Master. Adult and Master have exhibited higher occurrence of sports injuries than Adolescent group (p < 0.05). Joint injuries were the most common sports-related injuries by all Brazilian jiu-jitsu practitioners. While female gender and exposure time constituted the most predictive variables for sports injury occurrence in Adolescent, graduation level was more associated with sports injuries occurrence in Adult. CONCLUSIONS: Joint injuries derived from combat demands were the main sports injury in all age categories of Brazilian jiu-jitsu. Master subjects presented a higher occurrence of clinical changes and retrospective musculoskeletal injuries in relation to other age groups. Female gender and exposure time constituted the main predictive factors in adolescent subjects, while graduation category was more directly associated with retrospective injury onset in the Adult group.

ULCERATIVE AND NECROTIZING GASTRITIS IN A CAPTIVE SLOTH (BRADYPUS VARIEGATUS, XENARTHRA, BRADYPODIDAE) DUE TO SEVERE PARASITISM WITH PARALEIURIS LOCCHII (NEMATODA, SPIROCERCIDAE).

This is the first reported case of lethal gastric parasitism by the nematode Paraleiuris locchii in a captive sloth ( Bradypus variegatus ). There were more than 600 parasites in the stomach of the sloth, associated with extensive areas of ulceration and necrosis. The animal developed emaciation, dehydration, and anemia that progressed to death.

Concomitant helminth infection downmodulates the Vaccinia virus-specific immune response and potentiates virus-associated pathology.

The aim of this work was to elucidate the immunopathological mechanisms of how helminths may influence the course of a viral infection, using a murine model. Severe virulence, a relevant increase in the virus titres in the lung and a higher mortality rate were observed in Ascaris and Vaccinia virus (VACV) co-infected mice, compared with VACV mono-infected mice. Immunopathological analysis suggested that the ablation of CD8+ T cells, the marked reduction of circulating CD4+ T cells producing IFN-γ, and the robust pulmonary inflammation were associated with the increase of morbidity/mortality in co-infection and subsequently with the negative impact of concomitant pulmonary ascariasis and respiratory VACV infection for the host. On the other hand, when evaluating the impact of the co-infection on the parasitic burden, co-infected mice presented a marked decrease in the total number of migrating Ascaris lung-stage larvae in comparison with Ascaris mono-infection. Taken together, our major findings suggest that Ascaris and VACV co-infection may potentiate the virus-associated pathology by the downmodulation of the VACV-specific immune response. Moreover, this study provides new evidence of how helminth parasites may influence the course of a coincident viral infection.

Sports injuries in soccer according to tactical position: a retrospective survey / Lesões desportivas no futebol segundo posicionamento tático de jogo: análise por inquérito retrospectivo

Abstract Introduction: In soccer, the players' positions have been associated with specific functional overload, which may cause sports injuries. Objective: To investigate the occurrence and characterize sport injuries according to soccer player position. Methods: 232 male soccer players (129 professionals and 103 amateurs) from different sport teams in Mato Grosso do Sul, Brazil, were distributed in groups according to their soccer player position. Besides anthropometric characteristics, sports injuries were registered by using a referred morbidity survey. The occurrence of injuries was analyzed by means of the Goodman Test. Logistic regression models were used to investigate the relationship between different risk factors and the occurrence/-recurrence of sports injuries. Results: Forwards showed higher occurrence rates of sport injuries than other soccer position groups. Joint injuries in lower limbs constituted the most frequent registered cases. Muscle injuries in the back region were the most registered sports injuries among midfielders, while muscle damages in lower limbs were the primary injuries registered for other line positions. In the etiologic context, contact was the main cause of sports injuries in all groups. Most athletes (195) reported recurrence of sports injuries. Conclusion: The occurrence of sports injuries was higher among forwards. Traumatic joint and muscle injuries were the most prevalent registers in all line positions.

Resumo Introdução: No futebol, o posicionamento tático de jogo está relacionado com atribuições funcionais específicas, que podem predispor à ocorrência de lesões desportivas. Objetivo: Analisar a ocorrência e caracterizar as lesões desportivas típicas do futebol, relacionando-as com a posição tática de jogo. Métodos: 232 jogadores de futebol do sexo masculino, sendo 129 profissionais e 103 amadores das categorias de base de diferentes equipes desportivas do Mato Grosso do Sul, foram distribuídos em grupos, conforme o posicionamento tático de jogo. Além da caracterização antropométrica, foram tomadas informações sobre lesões desportivas por meio de inquérito de morbidade referida. A ocorrência de lesões foi analisada por meio de teste de Goodman. Modelos de regressão logística foram utilizados para investigar a relação entre diferentes fatores de risco, ocorrência e recidiva de lesão. Resultados: O grupo de Atacantes mostrou as maiores taxas de ocorrência de lesões. Lesões de joelho e tornozelo/pé consistiram nos principais registros articulares entre jogadores de linha, enquanto que afecções de punho e mão foram as mais comuns entre goleiros. As ocorrências musculares de localização lombar foram as mais reportadas por Meias, enquanto as afecções em membros inferiores foram as mais relatadas por laterais, zagueiros, volantes e atacantes. Quanto ao mecanismo etiológico, circunstâncias de trauma prevaleceram em todos os grupos. A maioria dos atletas (195) apresentou recidivas de lesões. Conclusão: A ocorrência de lesões desportivas foi maior entre atacantes. As lesões articulares traumáticas e musculares constituíram-se como os registros mais predominantes em todas as posições de linha.

Multiple Exposures to Ascaris suum Induce Tissue Injury and Mixed Th2/Th17 Immune Response in Mice.

Ascaris spp. infection affects 800 million people worldwide, and half of the world population is currently at risk of infection. Recurrent reinfection in humans is mostly due to the simplicity of the parasite life cycle, but the impact of multiple exposures to the biology of the infection and the consequences to the host's homeostasis are poorly understood. In this context, single and multiple exposures in mice were performed in order to characterize the parasitological, histopathological, tissue functional and immunological aspects of experimental larval ascariasis. The most important findings revealed that reinfected mice presented a significant reduction of parasite burden in the lung and an increase in the cellularity in the bronchoalveolar lavage (BAL) associated with a robust granulocytic pulmonary inflammation, leading to a severe impairment of respiratory function. Moreover, the multiple exposures to Ascaris elicited an increased number of circulating inflammatory cells as well as production of higher levels of systemic cytokines, mainly IL-4, IL-5, IL-6, IL-10, IL-17A and TNF-α when compared to single-infected animals. Taken together, our results suggest the intense pulmonary inflammation associated with a polarized systemic Th2/Th17 immune response are crucial to control larval migration after multiple exposures to Ascaris.