Small subunit ribosomal metabarcoding reveals extraordinary trypanosomatid diversity in Brazilian bats

Background: Bats are a highly successful, globally dispersed order of mammals that occupy a wide array of ecological niches. They are also intensely parasitized and implicated in multiple viral, bacterial and parasitic zoonoses. Trypanosomes are thought to be especially abundant and diverse in bats. In this study, we used 18S ribosomal RNA metabarcoding to probe bat trypanosome diversity in unprecedented detail. Methodology/Principal Findings: Total DNA was extracted from the blood of 90 bat individuals (17 species) captured along Atlantic Forest fragments of Espírito Santo state, southeast Brazil. 18S ribosomal RNA was amplified by standard and/or nested PCR, then deep sequenced to recover and identify Operational Taxonomic Units (OTUs) for phylogenetic analysis. Blood samples from 34 bat individuals (13 species) tested positive for infection by 18S rRNA amplification. Amplicon sequences clustered to 14 OTUs, of which five were identified as Trypanosoma cruzi I, T. cruzi III/V, Trypanosoma cruzi marinkellei, Trypanosoma rangeli, and Trypanosoma dionisii, and seven were identified as novel genotypes monophyletic to basal T. cruzi clade types of the New World. Another OTU was identified as a trypanosome like those found in reptiles. Surprisingly, the remaining OTU was identified as Bodo saltans–closest non-parasitic relative of the trypanosomatid order. While three blood samples featured just one OTU (T. dionisii), all others resolved as mixed infections of up to eight OTUs. Conclusions/Significance: This study demonstrates the utility of next-generation barcoding methods to screen parasite diversity in mammalian reservoir hosts. We exposed high rates of local bat parasitism by multiple trypanosome species, some known to cause fatal human disease, others non-pathogenic, novel or yet little understood. Our results highlight bats as a long-standing nexus among host-parasite interactions of multiple niches, sustained in part by opportunistic and incidental infections of consequence to evolutionary theory as much as to public health. Author summary: Bats make up a mega-diverse, intensely parasitized order of volant mammals whose unique behavioural and physiological adaptations promote infection by a vast array of microorganisms. Trypanosomes stand out as ancient protozoan parasites of bats. As cryptic morphology, low parasitaemia and selective growth in culture have recurrently biased survey, we used 18S ribosomal RNA metabarcoding to resolve bat trypanosomatid diversity in Atlantic Forest fragments of southeast Brazil. Next to several unknown species, our deep sequence-based detection and assignment protocol recognized multiple known human-pathogenic trypanosomes, another linked to reptile hosts as well as a non-parasitic kinetoplastid in the blood of various phyllostomid bats. The striking permissivity exposed here, in a region where bat trypanosomes recently featured in a fatal case of Chagas disease, compels further research on bats’ role in the dispersal and spill-over of various microorganisms among humans and wildlife

High-Current Double Pulse ECT Technique for Inspection of Ferromagnetic Materials

The detection of surface cracks of conductive materials that have a magnetic permeability higher than μ0, are usually made using the Magnetic Flux Leakage (MFL) technique. It requires the saturation of the specimen so that some magnetic flux lines escape the material when a defect is present. However, saturating the material can be very power consuming and if there is motion involved, eddy currents induced due to motion decrease or even null this method sensitivity as speed increases, which can be a disadvantage in cases such as railroad inspection. This work proposes a new technique to inspect the surface of ferromagnetic materials based on eddy currents. It is denominated high-current double pulse (Hi-CDP) ECT. The technique creates two consecutive pulses of currents (up to 1500 A) in a coil in the vicinity of the sample. Fig. 1 shows the simulation model used and the corresponding magnetic flux obtained in a point in the axis of a pancake coil (25 turns, id=15 mm, od=25 mm, heigh=10 mm), and in the vicinity of the sample material. The first pulse (starts at 0.1 ms) saturates the material, making it behave almost like a non-ferromagnetic material. The second pulse starts at 0.25 ms when the maximum current of the first pulse occurs (when the material is most saturated). When the second pulse occurs, eddy currents are induced. As the material is saturated, the ferromagnetic properties almost do not interfere with penetration depth and distribution of eddy currents, making it suitable for eddy current testing. Fig. 2 shows the derivative of the magnetic field obtained in a point located between the windings of the coil and the sample material, for a case without defect and in the presence of two similar defects with different depths (0.5 mm and 1.5 mm deep) in the vicinity of the point. It is possible to observe that the second current peak contains a perturbation that is different according to the defect. The signal derivative was chosen in order to distinguish the MFL from eddy currents perturbations

Characterization of Complex Defects Using Eddy Currents with Uniform Field Probes and Giant Magneto-Resistor (GMR) Detectors

Some defects in metallic plates present complex forms that difficult their geometric characterization. This is the case of defects with ramifications where the eddy currents cannot penetrate some concavity areas. The illumination of the material under test with a coherent excitation field presents some advantages, such as the possibility of applying the excitation into different directions, thus obtaining different patterns that may be correlated to increase the definition of the acquired signals. The generation of the coherent excitation is obtained by using a planar coil that produces a uniform excitation field inside a given area. It is not easy to correlate the magnetic field patterns, obtained by using single component giant magneto-resistor (GMR) sensors, with the real geometry of the defects. The interpretation of the measured data is much easier if the data are inversed to obtain the geometry of the eddy current lines inside the conductor. The inversion process was performed using the discrete Fourier transform of the field data and of the elementary dipole current kernel. The inversion was followed by Tikhonov regularization and automated determination of the regularization parameter. Results obtained for a defect with three linear segments in a star configuration are depicte

La Importancia de la Radiología Forense

Resumen La identificación de los cadáveres es uno de los objetivos más importantes de la necropsia en grandes desastres, en este caso debido a una explosión, los cadáveres pueden presentar múltiples daños, mutilaciones, cuerpos quemados, restos de materiales de la explosión, tales como productos químicos, cuerpos extraños y objetos metálicos del componente desencadenante entre otros, lo que dificulta la aplicación de los métodos científicos más reconocidos, es aquí donde se utilizan los estudios radiológicos para identificar e individualizar los cadáveres mediante radiografías de la pelvis, identificación del sexo a través del orificio pélvico, en la radiografía del cráneo a través de las mastoides, estudio del carpograma, también de las diferentes señales particulares de cadáveres que pueden identificarse mediante un estudio radiológico para el proceso de identificación indiciaria, entre las señales particulares encontramos, prótesis ortopédicas, fracturas antiguas con formación de callos óseos, vértebras fusionadas, variantes anatómicas, vestimenta y artículos personales presentes en los cuerpos, las características individuales descritas por la familia o las personas cercanas a las víctimas y las encontradas durante el examen corporal, la identificación fehaciente utilizando métodos científicos, como la comparación de huellas digitales, la comparación dental y la comparación genética ácido desoxirribonucleico (ADN), (Cruz,2019 ).Summary The identification of the corpses is one of the most important objectives of the autopsy in major disasters, in this case due to an explosion, the corpses can present multiple damages, mutilations, burned bodies, remains of materials from the explosion, such as chemical products , foreign bodies and metallic objects of the triggering component among others, which makes it difficult to apply the most recognized scientific methods, it is here that radiological studies are used to identify and individualize the corpses by means of radiographs of the pelvis, identification of sex through the pelvic orifice, in the x-ray of the skull through the mastoids, study of the carpogram, also of the different particular signs of corpses that can be identified by means of a radiological study for the identification process, among the particular signs we find orthopedic prostheses, fractures old with bone callus formation os, fused vertebrae, anatomical variants, clothing and personal items present in the bodies, the individual characteristics described by the family or people close to the victims and those found during the body examination, reliable identification using scientific methods, such as the comparison of fingerprints, dental comparison and deoxyribonucleic acid (DNA) genetic comparison

Formação de professores em comunidades colaborativas no interior da Amazônia

This paper reports on a study that aimed at describing and comprehending the transformation process of a local academic community inside the Amazon – named Formazon – into a collaborative learning community which includes school teachers, professors, and future educators from different areas of knowledge. The data was collected in an 18-month period from records and audio transcripts of meetings and from interviews. The narrative research was used to analyze the data, understanding it just as Clandinin and Connely (2000) did, as a way to bring meaning to the lived experience and also to investigate it. Its analysis aimed at answering the following questions: How has the Formazon community been learning and transforming itself? How has it been shaping itself into a collaborative community? What are the evidences of this transformation? Results indicate that, in relation to Formazon, collaborative interaction regarding both the participants’ school practices as well as the researching of these practices and of the politics permeating them, has outlined a new form of continuous education in the group, based on reflecting on and investigating school practices. Constituting a community that studies, questions, and researches school practices, Formazon has empowered the professional development process of its participants by increasing emancipation and professional autonomy.Este artigo relata um estudo que objetivou descrever e compreender o processo de transformação de uma comunidade acadêmica endógena do interior da Amazônia – o Formazon – em uma comunidade acadêmica colaborativa que integra professores escolares, professores acadêmicos e futuros professores de diferentes áreas do conhecimento. Os dados foram coletados a partir de registros de reuniões, entrevistas e transcrições de áudios dos encontros durante 18 meses, e sua análise buscou responder às seguintes indagações: Como a comunidade Formazon vem aprendendo e se transformando? Como vem se constituindo em uma comunidade colaborativa? Quais os indícios dessa transformação? Os resultados indicam que a interlocução colaborativa que, no âmbito do Formazon, se estabeleceu acerca das práticas de ensino dos participantes, de pesquisas dessas práticas e das políticas que as permeiam delineou um novo tipo de formação contínua no grupo, baseado na reflexão e na investigação sobre as práticas educativas. Ao constituir-se em uma comunidade de estudo, problematização e pesquisa das práticas docentes, o Formazon empoderou o processo de desenvolvimento profissional dos participantes, pois incrementou a emancipação e a autonomia profissional

Spider venom administration impairs glioblastoma growth and modulates immune response in a non-clinical model.

Molecules from animal venoms are promising candidates for the development of new drugs. Previous in vitro studies have shown that the venom of the spider Phoneutria nigriventer (PnV) is a potential source of antineoplastic components with activity in glioblastoma (GB) cell lines. In the present work, the effects of PnV on tumor development were established in vivo using a xenogeneic model. Human GB (NG97, the most responsive line in the previous study) cells were inoculated (s.c.) on the back of RAG-/- mice. PnV (100 µg/Kg) was administrated every 48 h (i.p.) for 14 days and several endpoints were evaluated: tumor growth and metabolism (by microPET/CT, using 18F-FDG), tumor weight and volume, histopathology, blood analysis, percentage and profile of macrophages, neutrophils and NK cells isolated from the spleen (by flow cytometry) and the presence of macrophages (Iba-1 positive) within/surrounding the tumor. The effect of venom was also evaluated on macrophages in vitro. Tumors from PnV-treated animals were smaller and did not uptake detectable amounts of 18F-FDG, compared to control (untreated). PnV-tumor was necrotic, lacking the histopathological characteristics typical of GB. Since in classic chemotherapies it is observed a decrease in immune response, methotrexate (MTX) was used only to compare the PnV effects on innate immune cells with a highly immunosuppressive antineoplastic drug. The venom increased monocytes, neutrophils and NK cells, and this effect was the opposite of that observed in the animals treated with MTX. PnV increased the number of macrophages in the tumor, while did not increase in the spleen, suggesting that PnV-activated macrophages were led preferentially to the tumor. Macrophages were activated in vitro by the venom, becoming more phagocytic; these results confirm that this cell is a target of PnV components. Spleen and in vitro PnV-activated macrophages were different of M1, since they did not produce pro- and anti-inflammatory cytokines. Studies in progress are selecting the venom molecules with antitumor and immunomodulatory effects and trying to better understand their mechanisms. The identification, optimization and synthesis of antineoplastic drugs from PnV molecules may lead to a new multitarget chemotherapy. Glioblastoma is associated with high morbidity and mortality; therefore, research to develop new treatments has great social relevance. Natural products and their derivatives represent over one-third of all new molecular entities approved by FDA. However, arthropod venoms are underexploited, although they are a rich source of new molecules. A recent in vitro screening of the Phoneutria nigriventer spider venom (PnV) antitumor effects by our group has shown that the venom significantly affected glioblastoma cell lines. Therefore, it would be relevant to establish the effects of PnV on tumor development in vivo, considering the complex neoplastic microenvironment. The venom was effective at impairing tumor development in murine xenogeneic model, activating the innate immune response and increasing tumor infiltrating macrophages. In addition, PnV activated macrophages in vitro for a different profile of M1. These activated PnV-macrophages have potential to fight the tumor without promoting tumorigenesis. Studies in progress are selecting the venom molecules with antitumor and immunomodulatory effects and trying to better understand their mechanisms. We aim to synthesize and carry out a formulation with these antineoplastic molecules for clinical trials. Spider venom biomolecules induced smaller and necrotic xenogeneic GB; spider venom activated the innate immune system; venom increased blood monocytes and the migration of macrophages to the tumor; activated PnV-macrophages have a profile different of M1 and have a potential to fight the tumor without promote tumorigenesis

Trade-Induced Skill Polarization

We study how the skill distribution in an economy responds to changes in wage gaps induced by trade integration. Using administrative data for Denmark (1993-2012), we conduct a two-step empirical analysis. In the first step, we predict changes in wage gaps that are triggered by exogenous trade shocks. In the second step, we estimate the impact of such changes on the skill distribution. The main results for Denmark show that both the average and the standard deviation of skills increase as a result of trade integration. We then extend our analysis to Portugal, using its administrative data (1993-2012), to shed light on the potential role the labor market and education policy may play in establishing the feedback effect of trade on the skill distribution. Finally, we provide a theoretical intuition to rationalize both sets of results

Blood Loss and Transcapillary Refill in Uncontrolled Treated Hemorrhage in Dogs

OBJETIVE: This study evaluated retroperitoneal hematomas produced by bilateral injury of iliac arteries (uncontrolled hemorrhage), blood volume loss, transcapillary refill, the effects of volume replacement on retroperitoneal bleeding and the hemodynamic changes with and without treatment. METHODS: Initial blood volume was determined with Tc99m-labelled red cells, and bleeding was evaluated by means of a portable scintillation camera positioned over the abdomen. Previously splenectomized mongrel dogs (16.8 ± 2.2 kg) were submitted to hemorrhage for 30 minutes and randomized into three groups: I - no treatment (n=7); II - treatment with 32 mL/kg of Lactated Ringer's for three to five minutes (n=7); and III - treatment with 4 mL/kg of 7.5% NaCl plus 6.0% dextran 70 for three to five minutes (n=7). They were studied for an additional 45 minutes. RESULTS: Volume replacement produced transitory recovery in hemodynamic variables, including mean pulmonary artery pressure, pulmonary capillary wedge pressure and cardiac index, with significant increase in dogs treated with 32 mL/kg of Lactated Ringer's and 7.5% NaCl plus 6.0% dextran 70 (p<0.001, against no treatment), along with a decrease (p<0.001) in the systemic vascular resistance index. Groups II and III had significant initial decreases in hematocrit and hemoglobin. The treated dogs (groups II and III) presented rebleeding, which was greater during treatment with 32 mL/kg of Lactated Ringer's (group II). CONCLUSIONS: Despite the rebleeding observed in treated groups, the utilization of hypertonic saline solution with dextran proved to be effective in the initial reanimation, producing evident transcapillary refill, while the Lactated Ringer's solution produced capillary extravasation and was ineffective in the initial volume replacement in this model of uncontrolled hemorrhage

The P2X7 receptor mediates Toxoplasma gondii Control in Macrophages through canonical NLRP3 inflammasome activation and reactive oxygen species production

Toxoplasma gondii (T. gondii) is the protozoan parasite that causes toxoplasmosis, a potentially fatal disease to immunocompromised patients, and which affects approximately 30% of the world\u27s population. Previously, we showed that purinergic signaling via the P2X7 receptor contributes to T. gondii elimination in macrophages, through reactive oxygen species (ROS) production and lysosome fusion with the parasitophorous vacuole. Moreover, we demonstrated that P2X7 receptor activation promotes the production of anti-parasitic pro-inflammatory cytokines during early T. gondii infection in vivo. However, the cascade of signaling events that leads to parasite elimination via P2X7 receptor activation remained to be elucidated. Here, we investigated the cellular pathways involved in T. gondii elimination triggered by P2X7 receptor signaling, during early infection in macrophages. We focused on the potential role of the inflammasome, a protein complex that can be co-activated by the P2X7 receptor, and which is involved in the host immune defense against T. gondii infection. Using peritoneal and bone marrow-derived macrophages from knockout mice deficient for inflammasome components (NLRP3-/-, Caspase-1/11-/-, Caspase-11-/-), we show that the control of T. gondii infection via P2X7 receptor activation by extracellular ATP (eATP) depends on the canonical inflammasome effector caspase-1, but not on caspase-11 (a non-canonical inflammasome effector). Parasite elimination via P2X7 receptor and inflammasome activation was also dependent on ROS generation and pannexin-1 channel. Treatment with eATP increased IL-1β secretion from infected macrophages, and this effect was dependent on the canonical NLRP3 inflammasome. Finally, treatment with recombinant IL-1β promoted parasite elimination via mitochondrial ROS generation (as assessed using Mito-TEMPO). Together, our results support a model where P2X7 receptor activation by eATP inhibits T. gondii growth in macrophages by triggering NADPH-oxidase-dependent ROS production, and also by activating a canonical NLRP3 inflammasome, which increases IL-1β production (via caspase-1 activity), leading to mitochondrial ROS generation