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Visceral leishmaniasis (VL) is the most severe clinical form of leishmaniasis, fatal if untreated. Vaccination is the most cost-effective approach to disease control; however, to date, no vaccines against human VL have been made available. This work examines the efficacy of a novel vaccine consisting of the Leishmania membrane protein KMP11, LEISH-F3+ (a recombinant fusion protein, composed of epitopes of the parasite proteins nucleoside hydrolase, sterol-24-c-methyltransferase, and cysteine protease B), and the sand fly salivary protein LJL143, in two dose ratios. The inclusion of the TLR4 agonist GLA-SE as an adjuvant, and the use of virosomes (VS) as a delivery system, are also examined. In a hamster model of VL, the vaccine elicited antigen-specific immune responses prior to infection with Leishmania infantum. Of note, the responses were greater when higher doses of KMP11 and LEISH-F3+ proteins were administered along with the GLA-SE adjuvant and/or when delivered within VS. Remarkably, hamsters immunized with the complete combination (i.e., all antigens in VS + GLA-SE) showed significantly lower parasite burdens in the spleen compared to those in control animals. This protection was underpinned by a more intense, specific humoral response against the KMP11, LEISH-F3+, and LJL143 antigens in vaccinated animals, but a significantly less intense antibody response to the pool of soluble Leishmania antigens (SLA). Overall, these results indicate that this innovative vaccine formulation confers protection against L. infantum infection, supporting the advancement of the vaccine formulation into process development and manufacturing and the conduction of toxicity studies towards future phase I human clinical trials.

Under perturbing conditions such as infection with Leishmania, a protozoan parasite living within the phagosomes in mammalian macrophages, cellular and organellar structures, and metabolism are dynamically regulated for neutralizing the pressure of parasitism. However, how modulations of the host cell metabolic pathways support Leishmania infection remains unknown. Herein, we report that lipid accumulation heightens the susceptibility of mice to L. donovani infection and promotes resistance to first‐line anti‐leishmanial drugs. Despite being pro‐inflammatory, the in vitro generated uninfected lipid‐laden macrophages (LLMs) or adipose‐tissue macrophages (ATMs) display lower levels of reactive oxygen and nitrogen species. Upon infection, LLMs secrete higher IL‐10 and lower IL‐12p70 cytokines, inhibiting CD4+ T cell activation and Th1 response suggesting a key modulatory role for intramacrophage lipid accumulation in anti‐leishmanial host defence. We, therefore, examined this causal relationship between lipids and immunomodulation using an in vivo high‐fat diet (HFD) mouse model. HFD increased the susceptibility to L. donovani infection accompanied by a defective CD4+ Th1 and CD8+ T cell response. The white adipose tissue of HFD mice displays increased susceptibility to L. donovani infection with the preferential infection of F4/80+CD11b+CD11c+ macrophages with higher levels of neutral lipids reserve. The HFD increased resistance to a first‐line anti‐leishmanial drug associated with a defective adaptive immune response. These data demonstrate that the accumulation of neutral lipids contributes to susceptibility to visceral leishmaniasis hindering host‐protective immune response and reducing the efficacy of antiparasitic drug therapies. [ABSTRACT FROM AUTHOR]

Apoptosis has been proposed as a key mechanism responsible for [CD4.sup.+] T cell depletion and immune dysfunction during HIV infection. We demonstrated that Q-VD-OPH, a caspase inhibitor, inhibits spontaneous and activation-induced death of T cells from SIV-infected rhesus macaques (RMs). When administered during the acute phase of infection, Q-VD-OPH was associated with (a) reduced levels of T cell death, (b) preservation of [CD4.sup.+]/[CD8.sup.+] T cell ratio in lymphoid organs and in the gut, (c) maintenance of memory [CD4.sup.+]T cells, and (d) increased specific [CD4.sup.+]T cell response associated with the expression of cytotoxic molecules. Although therapy was limited to the acute phase of infection, Q-VD-OPH-treated RMs showed lower levels of both viral load and cell-associated SIV DNA as compared with control SIV-infected RMs throughout the chronic phase of infection, and prevented the development of AIDS. Overall, our data demonstrate that Q-VD-OPH injection in SIV- infected RMs may represent an adjunctive therapeutic agent to control HIV infection and delaying disease progression to AIDS., Introduction Pathogenic HIV and SIV infections of humans and rhesus macaques (RMs), respectively, are associated with progressive [CD4.sup.+]T cell depletion and having a short half-life (1, 2). The regulation of [...]

A library of seventeen novel ether phospholipid analogues, containing 5-membered heterocyclic rings (1,2,3-triazolyl, isoxazolyl, 1,3,4-oxadiazolyl and 1,2,4-oxadiazolyl) in the lipid portion were designed and synthesized aiming to identify optimised miltefosine analogues. The compounds were evaluated for their in vitro antiparasitic activity against Leishmania infantum and Leishmania donovani intracellular amastigotes, against Trypanosoma brucei brucei and against different developmental stages of Trypanosoma cruzi. The nature of the substituents of the heterocyclic ring (tail) and the oligomethylene spacer between the head group and the heterocyclic ring was found to affect the activity and toxicity of these compounds leading to a significantly improved understanding of their structure–activity relationships. The early ADMET profile of the new derivatives did not reveal major liabilities for the potent compounds. The 1,2,3-triazole derivative 27 substituted by a decyl tail, an undecyl spacer and a choline head group exhibited broad spectrum antiparasitic activity. It possessed low micromolar activity against the intracellular amastigotes of two L. infantum strains and T. cruzi Y strain epimastigotes, intracellular amastigotes and trypomastigotes, while its cytotoxicity concentration (CC50) against THP-1 macrophages ranged between 50 and 100 μM. Altogether, our work paves the way for the development of improved ether phospholipid derivatives to control neglected tropical diseases.

The State Council, among the political institutions of the Brazilian Empire, is one of the most relevant in the process of understanding the conceptual roots of its foreign policy. In this object, the Pimenta Bueno Mission to Paraguay is an important example of how many practices became a regularity during the Brazilian international relation’s history were originated, by analyzing the political debates in a period of tendencies being formulated.

O Poli (Tereftalato de Etileno), mais conhecido pela sigla PET, é um dos polímeros de engenharia mais utilizado na produção embalagens, principalmente de frascos. A produção de garrafas de refrigerante, por exemplo, consiste em dois processos: injeção e sopro. É notável que o processo de injeção de PET seja afetado pela umidade ambiente, pois diversas injetoras possuem um secador no próprio silo de alimentação do equipamento. Desta forma, buscou-se avaliar se as condições ambientes também exerciam alguma influência no processo de sopro do PET. A análise foi realizada de forma amostral em dias diferentes, com condições ambientes diferentes e dentro da empresa Brasiaço Indústria e Comércio Ltda. Foi observado que apenas uma das condições ambientes afeta o processo de sopro de PET. Portanto, foi desenvolvido e proposto um método para correlacionar tal condição ambiente aos parâmetros do processo de sopro.

O texto trata da análise de como se dá o trabalho do pedagogo nos Institutos Federais de Ensino Ciencia e Tecnologia (IFs), Nesse sentido, partiu da seguinte questão de pesquisa: Como se apresenta o trabalho do pedagogo na educação profissional técnica e tecnológica dos Institutos Federais? Realizou-se por meio do levantamento de publicações de artigos sobre a atuação do pedagogo nos IFs em revistas dos próprios institutos entre o ano de criação dos institutos, 2008, até 2014 para uma análise do cenário de atuação desse profissional. Os resultados dessa análise apontaram que existe uma atuação do pedagogo técnico ligada aos processos de supervisão, gestão e administração escolar, exigindo do pedagogo a mobilização de amplos saberes que dão visão da totalidade educacional, envolvendo os conhecimentos didático-pedagógicos, administrativos e legais da educação.

This paper addresses the global efforts and reactions related to ecological accidents and the emergence of environmental law. The used methodology was literature research, involving variety of media. This paper aims to describe the historical remains and increasing complexity of expert activities. It addresses the discussions on the concept of law and ecologically balanced environment as part of human rights, and raises questions on the influences of capitalism, undeniable dynamism of the nature forces, and legal systems that recognize the nature as a subject of rights. It presents sustainability as a challenge and ethical imperative of humankind.

Canine leishmaniosis (CanL) prevention in the Mediterranean basin is considered essential to stop human zoonotic visceral leishmaniasis. In this context, vaccination of dogs is expected to have a significant impact in disease control. CaniLeish® (Virbac Animal Health) is one of a few CanL vaccines that are at this moment licensed in Europe. This vaccine contains purified excreted-secreted proteins of Leishmania having several antigens/immunogens with potential to influence serological response. Therefore, it is important to know if CaniLeish vaccination increased the diagnostic challenges associated with conventional serology, limiting the value of some antigens. To address this 20 dogs from a cohort of 35 healthy dogs that were vaccinated, maintained indoor for 1 month and then returned to their natural domiciles for 2 years. After this period, they were re-called to evaluate their clinical/parasitological condition and assess the evolution of seroreactivity against different antigens: soluble promastigote Leishmania antigens (SPLA), recombinant protein Leishmania infantum cytosolic peroxiredoxin, recombinant protein K39 (rK39), recombinant protein K28 and recombinant kinesin degenerated derived repeat using ELISA. Two years after vaccination all vaccinated non-infected animals were seropositive for SPLA. For the other antigens the serological profile was indistinguishable from non-infected animals. Moreover, vaccinated animals presented a characteristic relative serological profile, with higher normalized serological response to SPLA than rK39. This fact enabled to distinguish with sensitivity 92.3% and specificity 95.4%, vaccinated non-infected dogs from infected and non-infected dogs. Ultimately, relative serological profile enabled the detection of healthy vaccinated animals enabling more accurate serological surveys.

A presente pesquisa tem como objetivo central analisar a concepção de formação de professores presentes nos documentos oficiais que incidem sobre os Institutos Federais de Educação, Ciência e Tecnologia (IF), abordando o entendimento desta formação e seu locus no contexto histórico, político e social. Desta forma, a base lógica da presente investigação é a dialética, entendida a partir do materialismo histórico. Foi realizada uma pesquisa bibliográfica, além da busca e análise de documentos oficiais produzidos e publicados pela Secretaria de Educação Profissional e Tecnológica, que trazem concepções de formação de professores para os IF. A partir daí, se verifica que a concepção de formação de professores se aproxima das concepções pragmáticas e tecnicistas, com o trabalho em seu sentido reificado. Ponderase que, apesar de uma concepção alinhada às necessidades do sistema capitalista, a possibilidade de formação de professores nos IF abre espaço para o desenvolvimento de ações em torno de uma concepção de formação de professores como um pesquisador crítico em direção a uma educação emancipatória.

Choosing cultivars that require less chilling temperatures to overcome dormancy is crucial, for example, peaches that grow in subtropical areas, which is not an appropriate climate for them. Thus, alternative techniques are necessary to induce sprouting. Yet there are not many available products in market, restricting options for fruit growers to a reduced number of chemical molecules, which are often harm the health to applicators. Therefore, this study aimed to assess phenological cycles and productive performance of three peach cultivars, submitted to foliar nitrogen fertilizer and calcium nitrate to induce sprouting. This study took place at Experimental Orchard of School of Agriculture (FCA, UNESP), Botucatu, state of São Paulo. Three peach cultivars were evaluated: ‘Douradão’, ‘BRS Kampai’ and ‘BRS Rubimel’ by applying four doses of foliar nitrogen fertilizer (FNF) at 0, 1.25, 2.50 and 3.75%; associated with 4% calcium nitrate. A 3x4 factorial arrangement (3 cultivars X 4 FNF doses) in a randomized block design was used, with 4 replicates and 2 plants per plot, totalizing 96 plants. Results indicated that FNF positively affected cultivars development, providing wider sprouting, flowering and fruit set; consequently, greater production. Then FNF became a good alternative to induce sprouting in peaches at mild winter temperatures. Moreover, ‘BRS Rubimel’ presented high sowing percentage associated with low flowering and fruit set percentages, that is, low production mainly caused by its lack of adaptability to studied region.

Every year, up to 90,000 new cases of Visceral Leishmaniasis and 30,000 resultant deaths are estimated to occur worldwide. Such numbers give relevance to the continuous study of this complex form of the disease: a zoonosis and an anthroponosis; two known etiological agents (Leishmania infantum and L. donovani, respectively); with an estimated average ratio of 1 symptomatic per 10 asymptomatic individuals; and sometimes associated with atypical clinical presentations. This complexity, which results from a long co-evolutionary process involving vector-host, host-pathogen, and pathogen-vector interactions, is still not completely understood. The determinants of visceralization are not fully defined and the dichotomy resistance vs. susceptibility remains unsolved, translating into obstacles that delay the progress of global disease control. Inbred mouse models, with different susceptibility patterns to Leishmania infection, have been very useful in exploring this dichotomy. BALB/c and C57BL/6 mice were described as susceptible strains to L. donovani visceral infection, while SV/129 was considered resistant. Here, we used these three mouse models, but in the context of L. infantum infection, the other Leishmania species that cause visceral disease in humans, and dynamically compared their local and systemic infection-induced immune responses in order to establish a parallel and to ultimately better understand susceptibility vs. resistance in visceral leishmaniasis. Overall, our results suggest that C57BL/6 mice develop an intermediate “infection-phenotype” in comparison to BALB/c and SV/129 mouse strains, considering both the splenic parasite burden and the determined target organs weights. However, the immune mechanisms associated with the control of infection seem to be different in each mouse strain. We observed that both BALB/c and SV/129, but not C57BL/6 mice, show an infection-induced increase of splenic T follicular helper cells. On the other hand, differences detected in terms of CD21 expression by B cells early after infection, together with the quantified anti-Leishmania specific antibodies, suggest that SV/129 are faster than BALB/c and C57BL/6 mice in the assembly of an efficient B-cell response. Additionally, we observed an infection-induced increase in polyfunctional CD4+ T cells in the resistant SV/129 model, opposing an infection-induced increase in CD4+IL-10+ cells in susceptible BALB/c mice. Our data aligns with the observations reported for L. donovani infection and suggest that not only a single mechanism, but an interaction of several could be necessary for the control of this parasitic disease.

BackgroundObjective methods to measure physical activity (PA) can lead to better cross-cultural comparisons, monitoring temporal PA trends, and measuring the effect of interventions. However, when applying this technology in field-work, the accelerometer data processing is prone to methodological issues. One of the most challenging issues relates to standardizing total wear time to provide reliable data across participants. It is generally accepted that at least 4 complete days of accelerometer wear represent a week for adults. It is not known if this same assumption holds true for pregnant women.AimWe assessed the optimal number of days needed to obtain reliable estimates of overall PA and moderate-to-vigorous physical activity (MVPA) during the 2nd trimester in pregnancy using a raw triaxial wrist-worn accelerometer.MethodsCross-sectional analyses were carried out in the antenatal wave of the 2015 Pelotas (Brazil) Birth Cohort Study. Participants wore the wrist ActiGraph wGT3X-BT accelerometer for seven consecutive days. The daily average acceleration, which indicated overall PA, was measured as milli-g (mg), and time spent in MVPA (minutes/day) was analyzed in 5-minute bouts. ANOVA and Kruskal-Wallis tests were used to compare variability across days of the week. Bland-Altman plots and the Spearman-Brown Prophecy Formula were applied to determine the reliability coefficient associated with one to seven days of measurement.ResultsAmong 2,082 pregnant women who wore the accelerometer for seven complete days, overall and MVPA were lower on Sundays compared to other days of the week. Reliability of > = 0.80 to evaluate overall PA was reached with at least three monitoring days, whereas seven days were needed to estimate reliable measures of MVPA.ConclusionsOur findings indicate that obtaining one week of accelerometry in adults is appropriate for pregnant women, particularly to obtain differences on weekend days and reliably estimate overall PA and MVPA.

The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles (“MISEV”) guidelines for the field in 2014. We now update these “MISEV2014” guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities. Finally, a checklist is provided with summaries of key points.

The use of secretion pathways for effector molecule delivery by microorganisms is a trademark of pathogenesis. Leishmania extracellular vesicles (EVs) were shown to have significant immunomodulatory potential. Still, they will act in conjunction with other released parasite-derived products that might modify the EVs effects. Notwithstanding, the immunomodulatory properties of these non-vesicular components and their influence in the infectious process remains unknown. To address this, we explored both in vitro and in vivo the immunomodulatory potential of promastigotes extracellular material (EXO), obtained as a whole or separated in two different fractions: EVs or vesicle depleted EXO (VDE). Using an air pouch model, we observed that EVs and VDE induced a dose-dependent cell recruitment profile different from the one obtained with parasites, attracting significantly fewer neutrophils and more dendritic cells (DCs). Additionally, when we co-inoculated parasites with extracellular products a drop in cell recruitment was observed. Moreover, in vitro, while VDE (but not EVs) downregulated the expression of DCs and macrophages activation markers, both products were able to diminish the responsiveness of these cells to LPS. Finally, the presence of Leishmania infantum extracellular products in the inoculum promoted a dose-dependent infection potentiation in vivo, highlighting their relevance for the infectious process. In conclusion, our data demonstrate that EVs are not the only relevant players among the parasite exogenous products. This, together with the dose-dependency observed, opens new avenues to the comprehension of Leishmania infectious process. The approach presented here should be exploited to revisit existing data and considered for future studies in other infection models.

An amendment to this paper has been published and can be accessed via the original article.

Intracellular protozoan of the genus Leishmania, endemic in the Mediterranean basin, are the cause of cutaneous (CL), mucocutaneous (MCL), and visceral leishmaniasis (VL). A 75-year-old woman was admitted nine years after a second kidney transplant (KT), due to persistent pancytopenia and fever. She presented edema and erythema of the nose in the last two years and an exophytic nodular lesion located on the left arm, with areas of peripheral necrosis and central ulceration in the last 18 months. A bone marrow biopsy revealed features compatible with Leishmania amastigotes, and polymerase chain reaction test (PCR) for Leishmania infantum was positive. Moreover, biopsy and PCR for L. infantum of the cutaneous lesion on the patient’s left arm and nose and PCR from peripheral blood were positive. Thus, a diagnosis of CL, MCL, and VL was made, and liposomal amphotericin B was initiated, but the patient had an unfavorable outcome and died. This is the first report of a KT recipient presenting with the entire spectrum of leishmaniasis. In Portugal, this infection is rare—so a high degree of clinical suspicion is required for its diagnosis, especially in endemic regions, as visceral leishmaniasis is a potentially life-threatening infection.