Murine colon organoids as a novel model to study Trypanosoma cruzi infection and interactions with the intestinal epithelium

Chagas disease (CD) is a life-threatening illness caused by the parasite Trypanosoma cruzi (T. cruzi). With around seven million people infected worldwide and over 50,000 deaths per year, CD is a major public health issue in Latin America. The main route of transmission to humans is through a triatomine bug (vector-borne), but congenital and oral transmission have also been reported. The acute phase of CD presents mild symptoms but may develop into a long-lasting chronic illness, characterized by severely impaired cardiac, digestive, and neurological functions. The intestinal tissue appears to have a key role during oral transmission and chronic infection of CD. In this immune-privileged reservoir, dormant/quiescent parasites have been suggested to contribute to disease persistence, infection relapse, and treatment failure. However, the interaction between the intestinal epithelium and T. cruzi has not been examined in depth, in part, due to the lack of in vitro models that approximate to the biological and structural complexity of this tissue. Therefore, to understand the role played by the intestinal tissue during transmission and chronic infection, physiological models resembling the organ complexity are needed. Here we addressed this issue by establishing and characterizing adult stem cell-derived colonoid infection models that are clinically relevant for CD. 3D and 2D systems of murine intestinal organoids infected with T. cruzi Dm28c (a highly virulent strain associated with oral outbreaks) were analyzed at different time points by confocal microscopy. T. cruzi was able to invade and replicate in intestinal epithelial primary cells grown as intact organoids (3D) and monolayers (2D). The permissiveness to pathogen infection differed markedly between organoids and cell lines (primate and intestinal human cell lines). So far, this represents the first evidence of the potential that these cellular systems offer for the study of host-pathogen interactions and the discovery of effective anti-chagasic drugs.Agencia Nacional de Investigación e InnovaciónPasteur NetworkFOCEM (MERCOSUR Structural Convergence Fund

Modeling host-parasite interaction in chagas disease with murine intestinal organoids

Chagas disease (CD) is a potentially life-threatening illness caused by the parasite Trypanosoma cruzi (T. cruzi). With around seven million people infected worldwide and over 10,000 deaths per year, CD is a major public health issue in Latin America. The main route of transmission to humans is through a triatomine bug (vector-borne) and, to a minor extent, by blood transfusion, organ transplantation, laboratory accidents, congenitally and orally (food-borne). The acute phase of CD presents mild symptoms. If left untreated, it develops into a long-lasting chronic illness, characterized by severely impaired cardiac, digestive, and neurological functions. The intestinal tissue appears to have a key role during oral transmission and chronic infection of CD. In these immune-privileged reservoirs, dormant/quiescent parasites have been suggested to contribute to disease persistence, infection relapse, and treatment failure. However, the interaction between the intestinal epithelium and T. cruzi has not been examined in depth, in part, due to the lack of in vitro models resembling the biological and structural complexity of this organ. Therefore, to understand the pathophysiological role played by the intestinal tissue during transmission and chronic infection, we evaluated the progression of T. cruzi infection of murine colon organoids. In order to model CD, 3D and 2D systems of murine intestinal organoids were infected with T. cruzi Dm28c, a strain that has been associated with high virulence and oral outbreaks. At different time points, the presence and load of parasites in the organoids, as well as the host cell morphology were evaluated by confocal microscopy, and compared to those obtained with a classical infection model (Vero cells). We show that the parasite invades and replicates in intestinal epithelial primary cells grown as intact organoids (3D) and monolayers (2D). The permissiveness to pathogen infection differed markedly between the primary and the tumoral (Vero) cells. So far, this represents the first evidence of the potential of these nearly physiological cellular systems to study host-pathogen interaction for CD and/or for the future evaluation of anti-chagasic drugs.Agencia Nacional de Investigación e Innovación (ANII)FOCEM (MERCOSUR Structural Convergence Fund

Potencial de los organoides intestinales murinos para el estudio de la enfermedad de Chagas

Chagas disease (CD) is a potentially life-threatening illness caused by the parasite Trypanosoma cruzi (T. cruzi). With around seven million people infected worldwide and over 10,000 deaths per year, CD is a major public health issue in Latin America. The main route of transmission to humans is through a triatomine bug (vector-borne) and, to a minor extent, by blood transfusion, organ transplantation, laboratory accidents, congenitally and orally (food- borne). The acute phase of CD presents mild symptoms. If left untreated, it develops into a long-lasting chronic illness, characterized by severely impaired cardiac, digestive, and neurological functions. The intestinal tissue appears to have a key role during oral transmission and chronic infection of CD. In these immune-privileged reservoirs, dormant/quiescent parasites have been suggested to contribute to disease persistence, infection relapse, and treatment failure. However, the interaction between the intestinal epithelium and T. cruzi has not been examined in depth, in part, due to the lack of in vitro models resembling the biological and structural complexity of this organ. Therefore, to understand the pathophysiological role played by the intestinal tissue during transmission and chronic infection, we evaluated the progression of T. cruzi infection of murine colon organoids. In order to model CD, 3D and 2D systems of murine intestinal organoids were infected with T. cruzi Dm28c, a strain that has been associated with high virulence and oral outbreaks. At different time points, the presence and load of parasites in the organoids, as well as the host cell morphology were evaluated by confocal microscopy, and compared to those obtained with a classical infection model (Vero cells). We show that the parasite invades and replicates in intestinal epithelial primary cells grown as intact organoids (3D) and monolayers (2D). The permissiveness to pathogen infection differed markedly between the primary and the tumoral (Vero) cells. So far, this represents the first evidence of the potential of these nearly physiological cellular systems to study host-pathogen interaction for CD and/or for the future evaluation of anti-chagasic drugs.Agencia Nacional de Investigación e Innovación (ANII

Impact of COVID-19 on cardiovascular testing in the United States versus the rest of the world

Objectives: This study sought to quantify and compare the decline in volumes of cardiovascular procedures between the United States and non-US institutions during the early phase of the coronavirus disease-2019 (COVID-19) pandemic. Background: The COVID-19 pandemic has disrupted the care of many non-COVID-19 illnesses. Reductions in diagnostic cardiovascular testing around the world have led to concerns over the implications of reduced testing for cardiovascular disease (CVD) morbidity and mortality. Methods: Data were submitted to the INCAPS-COVID (International Atomic Energy Agency Non-Invasive Cardiology Protocols Study of COVID-19), a multinational registry comprising 909 institutions in 108 countries (including 155 facilities in 40 U.S. states), assessing the impact of the COVID-19 pandemic on volumes of diagnostic cardiovascular procedures. Data were obtained for April 2020 and compared with volumes of baseline procedures from March 2019. We compared laboratory characteristics, practices, and procedure volumes between U.S. and non-U.S. facilities and between U.S. geographic regions and identified factors associated with volume reduction in the United States. Results: Reductions in the volumes of procedures in the United States were similar to those in non-U.S. facilities (68% vs. 63%, respectively; p = 0.237), although U.S. facilities reported greater reductions in invasive coronary angiography (69% vs. 53%, respectively; p < 0.001). Significantly more U.S. facilities reported increased use of telehealth and patient screening measures than non-U.S. facilities, such as temperature checks, symptom screenings, and COVID-19 testing. Reductions in volumes of procedures differed between U.S. regions, with larger declines observed in the Northeast (76%) and Midwest (74%) than in the South (62%) and West (44%). Prevalence of COVID-19, staff redeployments, outpatient centers, and urban centers were associated with greater reductions in volume in U.S. facilities in a multivariable analysis. Conclusions: We observed marked reductions in U.S. cardiovascular testing in the early phase of the pandemic and significant variability between U.S. regions. The association between reductions of volumes and COVID-19 prevalence in the United States highlighted the need for proactive efforts to maintain access to cardiovascular testing in areas most affected by outbreaks of COVID-19 infection

O terrorismo e a intervenção da União Europeia na sua prevenção e repressão

Dissertação de mestrado em Direito da União EuropeiaEsta dissertação foi elaborada no âmbito do Mestrado em Direito da União Europeia. Neste documento apresentam-se e problematizam-se, à luz da bibliografia da especialidade, questões relacionadas com o terrorismo. Como temática de aprofundamento aborda-se O Terrorismo e a Intervenção da União Europeia, visando-se uma reflexão fundamentada sobre a sua Prevenção e Repressão. Neste sentido, foi importante o aprofundamento dos fundamentos e de toda a análise efetuada, subjacentes à temática, que serviu para analisar e refletir que tipo de tratamento, importância e combate a União Europeia tem tido para com a problemática do terrorismo. Esta exposição está dividida em cinco capítulos chave. No primeiro capítulo elaborou-se um enquadramento histórico-cronológico do terrorismo, desde o seu “possível” nascimento, à própria formação de grupos terroristas, como Al-Qaeda, e o sinal de mudança com o 11 de setembro de 2001. No segundo capítulo a análise centra-se na tentativa de encontrar uma definição consistente e realista para o terrorismo, tendo por base a legislação existente e a sua análise doutrinal. O terceiro capítulo reúne o seu estudo na criação e evolução do Espaço de Liberdade, Segurança e Justiça da União Europeia, da cooperação judiciária em matéria penal, que permitiu a coordenação, a colaboração e, acima de tudo, a união com o objetivo de combater este tipo de crime dos vários órgãos e organismos e do importantíssimo Princípio do Reconhecimento Mútuo. O quarto capítulo fala-nos do financiamento terrorista. O financiamento deste tipo de fenómeno permite muito mais facilmente a sua proliferação. Procuramos evidenciar quais as sanções legais para quem financia, direta ou indiretamente o terrorismo. O último capítulo procura refletir como combater o terrorismo. Foi realizada uma análise aprofundada sobre as soluções jurídicas de cunho europeu no que concerne à prevenção e repressão e consequentes implicações do terrorismo no ordenamento jurídico nacional. Neste cenário, considerando os contributos da bibliografia da especialidade, destacamos o facto de que a luta contra o terrorismo só acontecerá através da educação. Educando para a sensibilidade, pela preocupação e respeito pelo outro, pela diferença do outro, pela preocupação do outro. Neste contexto, importa que todos os Estados-Membros se unam em detrimento de um esforço coletivo, por aqueles e por outros que ainda não foram fustigados por esse crime.This dissertation was elaborated under the Master in European Union Law. During this paper, we will discuss the problematic of Terrorism and the issues related with it in the light of its current specialized literature. Moreover, we will give increased focus on the analysis of Terrorism and the intervention of European Union, aiming for a rational reflection on its Prevention and Suppression. In this sense, it was very important to deepen the fundamental issues and the analysis that is underlined in the theme, which served to study and reflect on what kind of treatment, significance and combat the European Union has had with the problematic of Terrorism. This presentation is divided into five key chapters. In the first chapter, an historical-chronological framework of Terrorism was elaborated, from its “conceivable” birth, to the very formation of terrorist groups, such as Al-Qaeda, and the sign of change that came with September 11, 2001. In the second chapter, the analysis focuses on trying to find a consistent and realistic definition of Terrorism, based on existing legislation and its doctrinal analysis. The third chapter brings together the study of the establishment and evolution of the European Union's area of freedom, security and justice, and of the judicial cooperation in criminal matters, which has enabled the coordination, collaboration and, above all, unity with the aim of battling this type of crime by the various organs and agencies, and the fundamental Principle of Mutual Recognition. The fourth chapter examines the sponsoring and financing of terrorism. Funding for this kind of phenomenon makes it much easier to proliferate and, for that reason, we try to highlight the legal sanctions for those who finance terrorism, be it directly or indirectly. The last chapter focuses on how to combat terrorism. For that reason, it was carried out an in-depth analysis of the European legal solutions regarding prevention and repression, and the consequent implications of Terrorism on the national legal system. In this scenario, and considering the contributions of the specialized literature on this theme, we highlight the fact that the battle against terrorism will only see actual results through education. Educating for sensitivity, concern and respect for the other, for the difference of the other, and for the empathy for the other. In Portugal, there is a law to fight terrorism and we have not yet suffered any attacks, but it does not imply that all Member States will unite at the expense of a collective effort for those who have already been confronted with this type of criminal act

Identification and relative quantification of 3-nitrotyrosine residues in fibrinogen nitrated in vitro and fibrinogen from ischemic stroke patient plasma using LC-MS/MS

Ischemic stroke is one of the leading causes of death and disability worldwide. This acute vascular event interferes with blood supply to the brain and induces a burst of free radicals such as nitric oxide and superoxide, producing peroxynitrite, a precursor of strong nitrating agents. Fibrinogen is one of the most abundant plasma proteins; it plays a role in the hemostatic system, mediating clot formation, which can be affected by nitrotyrosine formation. We hypothesized that nitration of fibrinogen by ONOOH and ONOOCO radical products could be one of the early events of the ischemic stroke, and protein-bound 3-nitrotyrosine could be a potential biomarker for diagnosis and/or prognosis of this condition. A targeted mass spectrometry approach was developed to analyze the nitration of fibrinogen and its association with ischemic stroke. First, a comprehensive mapping of 3-nitrotyrosine locations and their relative quantification was performed by LC-MS/MS, using in vitro nitrated fibrinogen samples. Twenty different 3-nitrotyrosine residues were identified on fibrinogen nitrated in vitro, varying with the peroxynitrite tofibrinogen molar ratio used. Nine tyrosine residues that were consistently modified at different treatment ratios were chosen to perform a targeted LC-MS/MS analysis in clinical samples. Enriched fibrinogen fractions from clinical samples from 24 ischemic stroke and 12 patients with non-inflammatory conditions were analysed with this method. Three of the nine tyrosine residues analysed (βY452, βY475 and γY380) showed a significant difference between the ischemic stroke and non-inflammatory disease groups. ROC curve analysis suggested an association of these residues either individually or in combination with ischemic stroke. Different tyrosine nitration patterns were also observed in fibrinogen modified in vitro and in vivo, suggesting differences in the nitration process in these situations. This is the first study showing a putative association between the nitration profile of specific tyrosine residues in human fibrinogen and ischemic stroke. [Abstract copyright: Copyright © 2021 Elsevier Inc. All rights reserved.

Organoides intestinales: una herramienta versátil para el estudio in vitro de patologías del epitelio intestinal

Los organoides intestinales son estructuras multicelulares tridimensionales que derivan de células madre y tienen la capacidad de auto-organizarse. Recrean varios aspectos de la morfología, composición celular y fisiología del intestino, constituyendo modelos del epitelio intestinal de mayor relevancia que las líneas celulares tradicionales. El objetivo de este trabajo consistió en implementar el cultivo de organoides intestinales murinos, bovinos y ovinos, a partir de células madre adultas. Para posteriormente emplearlos como herramientas de reducción del uso de animales de experimentación y para el estudio de patologías asociadas al epitelio intestinal.Agencia Nacional de Investigación e Innovació

¿El COVID-19 cambió la percepción de la naturaleza?: Un análisis de las representaciones sociales durante la pandemia en Argentina.

La pandemia de COVID-19 desató discusiones sobre la relación sociedad-naturaleza y sobre la crisis civilizatoria, consecuencia de la dominación antrópica del ambiente. Este trabajo parte de la hipótesis de que las experiencias singulares producidas por la pandemia alteran las representaciones acerca de la naturaleza y generan nuevas perspectivas.Nos propusimos analizar estas representaciones y sus transformaciones. Nos preguntamos cómo es concebida la naturaleza, si existe una dualidad percibida entre naturaleza y sociedad o no. Analizamos los cambios generados por el escenario pandémico, con el énfasis puesto en las percepciones y comportamientos y, en especial, en la reflexión sobre esta relación en la pospandemia.Este trabajo es parte de la investigación de la Red del Estudio Nacional Colaborativo de Representaciones sobre la Pandemia en Argentina (Encrespa), y tiene como objetivos comprender las transformaciones en curso, producir conocimiento aplicable a políticas de sostenibilidad y reflexionar sobre el rol de la producción de conocimiento científico ante los desafíos de la pospandemia.The Covid-19 pandemic sparked discussions about the relationship between society and nature and about the civilizational crisis as a consequence of the anthropic domination of the environment. This article stems from the hypothesis that the unique experiences produced by the pandemic alter representations about nature and generate new perspectives. We set out to analyze these representations and their transformations. We ask how nature is conceived; whether there exists a perceived duality between nature-society or not. We analyze the changes generated by the pandemic scenario, emphasizing perceptions and behaviors and, especially, in the reflection on this relationship in the post-pandemic. This work is part of the investigation of the Network of National Collaborative Research of Representations on the Pandemic in Argentina (ENCRESPA in spanish), and its objectives are to comprehend the current transformations, produce knowledge applicable to sustainability policies, and reflect on the role of the production of scientific knowledge in the face of the post-pandemic challenges.Fil: Van Aert, Peter Louisa Petrus. Universidad Nacional de Tierra del Fuego. Instituto de Cultura, Sociedad y Estado; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas; ArgentinaFil: Calvelo, Laura Inés. Universidad Nacional de Avellaneda. Departamento de Articulac. Transversal de Comunic. y Tecn. de la Info.; ArgentinaFil: Datri, Andrea Marina. Universidad Nacional de La Pampa; ArgentinaFil: Duarte de Medeiros, Dulcinea. Universidad Nacional de Tierra del Fuego. Instituto de Cultura, Sociedad y Estado; ArgentinaFil: Mansilla, Paula Romina. Universidad Nacional de Tierra del Fuego. Instituto de Ciencias Polares, Recursos Naturales y Ambiente; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas; ArgentinaFil: Pescader, Carlos Alberto. Universidad Nacional del Comahue; ArgentinaFil: Rojas, Facundo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Argentino de Nivología, Glaciología y Ciencias Ambientales. Provincia de Mendoza. Instituto Argentino de Nivología, Glaciología y Ciencias Ambientales. Universidad Nacional de Cuyo. Instituto Argentino de Nivología, Glaciología y Ciencias Ambientales; ArgentinaFil: Wagner, Lucrecia Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Argentino de Nivología, Glaciología y Ciencias Ambientales. Provincia de Mendoza. Instituto Argentino de Nivología, Glaciología y Ciencias Ambientales. Universidad Nacional de Cuyo. Instituto Argentino de Nivología, Glaciología y Ciencias Ambientales; Argentin

Plant Microbiome Engineering: Hopes or Hypes

Rhizosphere microbiome is a dynamic and complex zone of microbial communities. This complex plant-associated microbial community, usually regarded as the plant&rsquo;s second genome, plays a crucial role in plant health. It is unquestioned that plant microbiome collectively contributes to plant growth and fitness. It also provides a safeguard from plant pathogens, and induces tolerance in the host against abiotic stressors. The revolution in omics, gene-editing and sequencing tools have somehow led to unravel the compositions and latent interactions between plants and microbes. Similarly, besides standard practices, many biotechnological, (bio)chemical and ecological methods have also been proposed. Such platforms have been solely dedicated to engineer the complex microbiome by untangling the potential barriers, and to achieve better agriculture output. Yet, several limitations, for example, the biological obstacles, abiotic constraints and molecular tools that capably impact plant microbiome engineering and functionality, remained unaddressed problems. In this review, we provide a holistic overview of plant microbiome composition, complexities, and major challenges in plant microbiome engineering. Then, we unearthed all inevitable abiotic factors that serve as bottlenecks by discouraging plant microbiome engineering and functionality. Lastly, by exploring the inherent role of micro/macrofauna, we propose economic and eco-friendly strategies that could be harnessed sustainably and biotechnologically for resilient plant microbiome engineering