In vivo tracking and immunological properties of pulsed porcine monocyte-derived dendritic cells

Cellular therapies using immune cells and in particular dendritic cells (DCs) are being increasingly applied in clinical trials and vaccines. Their success partially depends on accurate delivery of cells to target organs or migration to lymph nodes. Delivery and subsequent migration of cells to regional lymph nodes is essential for effective stimulation of the immune system. Thus, the design of an optimal DC therapy would be improved by optimizing technologies for monitoring DC trafficking. Magnetic resonance imaging (MRI) represents a powerful tool for non-invasive imaging of DC migration in vivo. Domestic pigs share similarities with humans and represent an excellent animal model for immunological studies. The aim of this study was to investigate the possibility using pigs as models for DC tracking in vivo. Porcine monocyte derived DC (MoDC) culture with superparamagnetic iron oxide (SPIO) particles was standardized on the basis of SPIO concentration and culture viability. Phenotype, cytokine production and mixed lymphocyte reaction assay confirmed that porcine SPIO-MoDC culture were similar to mock MoDCs and fully functional in vivo. Alike, similar patterns were obtained in human MoDCs. After subcutaneous inoculation in pigs, porcine SPIO-MoDC migration to regional lymph nodes was detected by MRI and confirmed by Perls staining of draining lymph nodes. Moreover, after one dose of virus-like particles-pulsed MoDCs specific local and systemic responses were confirmed using ELISPOT IFN-γ in pigs. In summary, the results in this work showed that after one single subcutaneous dose of pulsed MoDCs, pigs were able to elicit specific local and systemic immune responses. Additionally, the dynamic imaging of MRI-based DC tracking was shown using SPIO particles. This proof-of-principle study shows the potential of using pigs as a suitable animal model to test DC trafficking with the aim of improving cellular therapies.We want to thank: Ferrán López, Rosa López, Zoraida Cervera, Pamela Martinez-Orellana, Tufaria Mussá, Massimiliano Baratelli, Diego Pérez, Sergio López from CRESA and José Luis Ruiz de la Torre and Javier Aceña (UAB) for farm and technical support; Jaume Martorell (Fundació Hospital Clínic Veterinari, UAB) for MRI support; Javier Domínguez (INIA) for the porcine antibodies; Antonio Lestuzzi, Michele Crisci and Raif Yucel for MR imaging support; Joaquim Segalés for anatomic pathology analysis; Mónica Pérez for immunohistochemical stainings; Aida Neira and Blanca Pérez for Perls staining; Eva Huerta y Marina Sibila for PCV2 PCR; David Andreu and Beatriz García de la Torre (Pompeu Fabra University, Barcelona), and Esther Blanco (CISA-INIA, Madrid), for the FMDV 3A peptide; Alicia Solórzano for critically reviewing the manuscript. This work was funded by the project AGL2010-22200-C02 of Spanish Ministry of Science and Innovation. PhD studies of Raquel Cabezón are funded by a doctoral FI fellowship from the Generalitat de Catalunya

10 anos do IPDMS: realizações, limites e desafios

This text it was writhed for two people associated of Instituto de Pesquisa, Direitos e Movimentos Sociais since the foundation. They built many spaces and composed the Secretaria Nacional for many years. Since this engagement and respecting their personal, professional and politics trajectories, intend to cast approaches about this decade of IPDMS history. Furthermore, they point out questions for a possible prospective balance. Therefore, provide a brief description of the institute’s creation anteroom, which drew from the source of networks articulations of student movement and popular advocacy, as well as popular education and action research networks. Esse texto é escrito por duas pessoas associadas ao Instituto de Pesquisa, Direitos e Movimentos Sociais desde sua fundação, que construíram inúmeros espaços do mesmo e estiveram à frente de sua Secretaria Nacional por alguns anos. Desde este engajamento e respeitando suas particulares trajetórias pessoais, profissionais e políticas, pretendem lançar olhares sobre esta década de história do IPDMS, bem como apontar questões para um possível balanço prospectivo. Para tanto, traçam uma breve descrição da antessala de criação do instituto, que bebeu na fonte de articulações em rede de movimento estudantil e advocacia popular, assim como redes de educação popular e de pesquisa ação

Sobre reaprender o significado do trabalho de base, reinventá-lo diante das atuais configurações da realidade e poder “arrancar alegria ao futuro”: Entrevista com Ayala Lindabeth Dias Ferreira

Entrevista com Ayala Lindabeth Dias Ferreira, realizada por Carla Benitez Martins e Luiz Otávio Ribas

Plasmacytoid dendritic cells of melanoma patients present exogenous proteins to CD4+ T cells after FcγRII-mediated uptake

Plasmacytoid dendritic cells (pDCs) contribute to innate antiviral immune responses by producing type I interferons. Although human pDCs can induce T cell responses upon viral infection, it remains unclear if pDCs can present exogenous antigens. Here, we show that human pDCs exploit FcγRII (CD32) to internalize antigen–antibody complexes, resulting in the presentation of exogenous antigen to T cells. pDCs isolated from melanoma patients vaccinated with autologous monocyte-derived peptide- and keyhold limpet hemocyanin (KLH)–loaded dendritic cells, but not from nonvaccinated patients or patients that lack a humoral response against KLH, were able to stimulate KLH-specific T cell proliferation. Interestingly, we observed that internalization of KLH by pDCs depended on the presence of serum from vaccinated patients that developed an anti-KLH antibody response. Anti-CD32 antibodies inhibited antigen uptake and presentation, demonstrating that circulating anti-KLH antibodies binding to CD32 mediate KLH internalization. We conclude that CD32 is an antigen uptake receptor on pDCs and that antigen presentation by pDCs is of particular relevance when circulating antibodies are present. Antigen presentation by pDCs may thus modulate the strength and quality of the secondary phase of an immune response

UM OLHAR EXPLORATÓRIO SOBRE OS RESÍDUOS DE SERVIÇOS DE SAÚDE PARA OS CURSOS DA ÁREA DA SAÚDE NUMA UNIVERSIDADE COMUNITÁRIA DO SUL DO BRASIL

This research aims a management system Waste Health Services - RSS - courses for health care from a community college, based on guidance provided by the Brazilian legislation linked to the health sector, the Environmental Management System, established by ISO 14001, and application of two questionnaires: one for collecting data from the students and one for faculty and staff. The research was exploratory and descriptive approaches used to integrate qualitative and quantitative analysis. It was found that 94% of students are unaware of the legislation concerning RSS and that 82% of teachers and officials do not know the destination of the waste. Other survey results demonstrated the need for more extensive and consistent information regarding RSS, generated in practical classes and community service. The knowledge of both the students and the faculty and staff, is focused and specific to each situation, there is a gap in the legislation and management of RSS. After the results were compiled systematic suggestions about Waste Management of Health Services, generated in teaching, which is a set of actions aiming at the quality and efficiency of academic practices, emphasizing environmental education. Esta pesquisa objetiva realizar uma investigação sobre os resíduos de serviços de saúde para os cursos da área da saúde numa universidade comunitária do sul do Brasil com base nas orientações fornecidas pela legislação brasileira vinculada ao setor de saúde e no Sistema de Gestão Ambiental, estabelecido pela NBR ISO 14001, para a proposição de um modelo para Gerenciamento de Resíduos de Serviços de Saúde. Para isto, foram aplicados dois questionários: um para coleta de dados dos alunos e outro para docentes e funcionários. A pesquisa classificou-se como exploratória descritiva e usou abordagens de integração entre análise qualitativa e quantitativa. Foi constatado que 94% dos alunos desconhecem a legislação referente aos Resíduos de Serviços de Saúde - RSS e que 82% dos docentes e funcionários não conhecem a destinação desses resíduos. Outros resultados da pesquisa demonstraram a necessidade de informações mais amplas e consistentes em relação aos RSS gerados nas aulas práticas e no atendimento à comunidade. O conhecimento das técnicas aplicadas tanto pelos alunos quanto pelos docentes e funcionários, focadas especificamente nas áreas de atuação, deixam a desejar no que diz respeito à legislação e ao gerenciamento dos RSS. Percebeu-se que esse assunto não é tratado pelos docentes, discentes e técnicos administrativos como relevante, embora, todos saibam da sua importância vital à saúde de cada sujeito envolvido. Após os resultados obtidos, foram elaboradas sugestões sistemáticas sobre a gestão de Resíduos de Serviços de Saúde gerados em atividades de ensino que se constitui num conjunto de ações visando à qualidade e à eficiência das práticas acadêmicas, enfatizando a educação ambiental

Post-mortem neuropathologic examination of a 5-case series of CAR T-cell treated patients

Introduction: Chimeric antigen receptor (CAR) T-cell therapy is a promising immunotherapy for the treatment of refractory hematopoietic malignancies. Adverse events are common, and neurotoxicity is one of the most important. However, the physiopathology is unknown and neuropathologic information is scarce.Materials and methods: Post-mortem examination of 6 brains from patients that underwent CAR T-cell therapy from 2017 to 2022. In all cases, polymerase chain reaction (PCR) in paraffin blocks for the detection of CAR T cells was performed.Results: Two patients died of hematologic progression, while the others died of cytokine release syndrome, lung infection, encephalomyelitis, and acute liver failure. Two out of 6 presented neurological symptoms, one with extracranial malignancy progression and the other with encephalomyelitis. The neuropathology of the latter showed severe perivascular and interstitial lymphocytic infiltration, predominantly CD8+, together with a diffuse interstitial histiocytic infiltration, affecting mainly the spinal cord, midbrain, and hippocampus, and a diffuse gliosis of basal ganglia, hippocampus, and brainstem. Microbiological studies were negative for neurotropic viruses, and PCR failed to detect CAR T -cells. Another case without detectable neurological signs showed cortical and subcortical gliosis due to acute hypoxic-ischemic damage. The remaining 4 cases only showed a mild patchy gliosis and microglial activation, and CAR T cells were detected by PCR only in one of them.Conclusions: In this series of patients that died after CAR T-cell therapy, we predominantly found non-specific or minimal neuropathological changes. CAR T-cell related toxicity may not be the only cause of neurological symptoms, and the autopsy could detect additional pathological findings

Low-density lipoprotein receptor-related protein 1 deficiency in cardiomyocytes reduces susceptibility to insulin resistance and obesity

Background: Low-density lipoprotein receptor-related protein 1 (LRP1) plays a key role in fatty acid metabolism and glucose homeostasis. In the context of dyslipemia, LRP1 is upregulated in the heart. Our aim was to evaluate the impact of cardiomyocyte LRP1 deficiency on high fat diet (HFD)-induced cardiac and metabolic alterations, and to explore the potential mechanisms involved. Methods: We used TnT-iCre transgenic mice with thoroughly tested suitability to delete genes exclusively in cardiomyocytes to generate an experimental mouse model with conditional Lrp1 deficiency in cardiomyocytes (TNT-iCre+-LRP1flox/flox). Findings: Mice with Lrp1-deficient cardiomyocytes (cm-Lrp1-/-) have a normal cardiac function combined with a favorable metabolic phenotype against HFD-induced glucose intolerance and obesity. Glucose intolerance protection was linked to higher hepatic fatty acid oxidation (FAO), lower liver steatosis and increased whole-body energy expenditure. Proteomic studies of the heart revealed decreased levels of cardiac pro-atrial natriuretic peptide (pro-ANP), which was parallel to higher ANP circulating levels. cm-Lrp1-/- mice showed ANP signaling activation that was linked to increased fatty acid (FA) uptake and increased AMPK/ ACC phosphorylation in the liver. Natriuretic peptide receptor A (NPR-A) antagonist completely abolished ANP signaling and metabolic protection in cm-Lrp1-/- mice. Conclusions: These results indicate that an ANP-dependent axis controlled by cardiac LRP1 levels modulates AMPK activity in the liver, energy homeostasis and whole-body metabolism

Habitable Zones in the Universe

Habitability varies dramatically with location and time in the universe. This was recognized centuries ago, but it was only in the last few decades that astronomers began to systematize the study of habitability. The introduction of the concept of the habitable zone was key to progress in this area. The habitable zone concept was first applied to the space around a star, now called the Circumstellar Habitable Zone. Recently, other, vastly broader, habitable zones have been proposed. We review the historical development of the concept of habitable zones and the present state of the research. We also suggest ways to make progress on each of the habitable zones and to unify them into a single concept encompassing the entire universe.Comment: 71 pages, 3 figures, 1 table; to be published in Origins of Life and Evolution of Biospheres; table slightly revise

Non-invasive ventilation in obesity hypoventilation syndrome without severe obstructive sleep apnoea

Background Non-invasive ventilation (NIV) is an effective form of treatment in patients with obesity hypoventilation syndrome (OHS) who have concomitant severe obstructive sleep apnoea (OSA). However, there is a paucity of evidence on the efficacy of NIV in patients with OHS without severe OSA. We performed a multicentre randomised clinical trial to determine the comparative efficacy of NIV versus lifestyle modification (control group) using daytime arterial carbon dioxide tension (PaCO2) as the main outcome measure. Methods Between May 2009 and December 2014 we sequentially screened patients with OHS without severe OSA. Participants were randomised to NIV versus lifestyle modification and were followed for 2 months. Arterial blood gas parameters, clinical symptoms, health-related quality of life assessments, polysomnography, spirometry, 6-min walk distance test, blood pressure measurements and healthcare resource utilisation were evaluated. Statistical analysis was performed using intention-to-treat analysis. Results A total of 365 patients were screened of whom 58 were excluded. Severe OSA was present in 221 and the remaining 86 patients without severe OSA were randomised. NIV led to a significantly larger improvement in PaCO2 of -6 (95% CI -7.7 to -4.2) mm Hg versus -2.8 (95% CI -4.3 to -1.3) mm Hg, (p<0.001) and serum bicarbonate of -3.4 (95% CI -4.5 to -2.3) versus -1 (95% CI -1.7 to -0.2 95% CI) mmol/L (p<0.001). PaCO2 change adjusted for NIV compliance did not further improve the inter-group statistical significance. Sleepiness, some health-related quality of life assessments and polysomnographic parameters improved significantly more with NIV than with lifestyle modification. Additionally, there was a tendency towards lower healthcare resource utilisation in the NIV group. Conclusions NIV is more effective than lifestyle modification in improving daytime PaCO2, sleepiness and polysomnographic parameters. Long-term prospective studies are necessary to determine whether NIV reduces healthcare resource utilisation, cardiovascular events and mortality