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Abstract Retinal degenerative diseases, including diabetic retinopathy (DR) and age-related macular degeneration (AMD), loom as threats to vision, causing detrimental effects on the structure and function of the retina. Central to understanding these diseases, is the compromised state of the blood-retinal barrier (BRB), an effective barrier that regulates the influx of immune and inflammatory components. Whether BRB breakdown initiates retinal distress, or is a consequence of disease progression, remains enigmatic. Nevertheless, it is an indication of retinal dysfunction and potential vision loss. The intricate intercellular dialogues among retinal cell populations remain unintelligible in the complex retinal milieu, under conditions of inflammation and oxidative stress. The retina, a specialized neural tissue, sustains a ceaseless demand for oxygen and nutrients from two vascular networks. The BRB orchestrates the exchange of molecules and fluids within this specialized region, comprising the inner BRB (iBRB) and the outer BRB (oBRB). Extracellular vesicles (EVs) are small membranous structures, and act as messengers facilitating intercellular communication in this milieu. EVs, both from retinal and peripheral immune cells, increase complexity to BRB dysfunction in DR and AMD. Laden with bioactive cargoes, these EVs can modulate the retinal microenvironment, influencing disease progression. Our review delves into the multifaceted role of EVs in retinal degenerative diseases, elucidating the molecular crosstalk they orchestrate, and their microRNA (miRNA) content. By shedding light on these nanoscale messengers, from their biogenesis, release, to interaction and uptake by target cells, we aim to deepen the comprehension of BRB dysfunction and explore their therapeutic potential, therefore increasing our understanding of DR and AMD pathophysiology.

Abstract Objectives Male infertility accounts for approximately 30% of cases of reproductive failure. The characterization of genetic variants using cytogenomic techniques is essential for the adequate clinical management of these patients. We aimed to conduct a cytogenetic investigation of numerical and structural rearrangements and a genomic study of Y chromosome microdeletions/microduplications in infertile men derived from a single centre with over 14 years of experience. Results We evaluated 151 infertile men in a transversal study using peripheral blood karyotypes and 15 patients with normal karyotypes through genomic investigation by multiplex ligation-dependent probe amplification (MLPA) or polymerase chain reaction of sequence-tagged sites (PCR-STS) techniques. Out of the 151 patients evaluated by karyotype, 13 presented chromosomal abnormalities: two had numerical alterations, and 11 had structural chromosomal rearrangements. PCR-STS detected a BPY2 gene region and RBMY2DP pseudogene region microdeletion in one patient. MLPA analysis allowed the identification of one patient with CDY2B_1 and CDY2B_2 probe duplications (CDY2B and NLGN4Y genes) and one patient with BPY2_1, BPY2_2, and BPY2_4 probe duplications (PRY and RBMY1J genes).

Abstract Malignant pleural mesothelioma (MPM) is an aggressive tumor with a poor prognosis. As the available therapeutic options show a lack of efficacy, novel therapeutic strategies are urgently needed. Given its T-cell infiltration, we hypothesized that MPM is a suitable target for therapeutic cancer vaccination. To date, research on mesothelioma has focused on the identification of molecular signatures to better classify and characterize the disease, and little is known about therapeutic targets that engage cytotoxic (CD8+) T cells. In this study we investigate the immunopeptidomic antigen-presented landscape of MPM in both murine (AB12 cell line) and human cell lines (H28, MSTO-211H, H2452, and JL1), as well as in patients’ primary tumors. Applying state-of-the-art immuno-affinity purification methodologies, we identify MHC I-restricted peptides presented on the surface of malignant cells. We characterize in vitro the immunogenicity profile of the eluted peptides using T cells from human healthy donors and cancer patients. Furthermore, we use the most promising peptides to formulate an oncolytic virus-based precision immunotherapy (PeptiCRAd) and test its efficacy in a mouse model of mesothelioma in female mice. Overall, we demonstrate that the use of immunopeptidomic analysis in combination with oncolytic immunotherapy represents a feasible and effective strategy to tackle untreatable tumors.

Abstract Background Neurofibromatosis type 1 (NF1) is an inherited neurocutaneous disorder associated with neurodevelopmental disorders including autism spectrum disorder (ASD). This condition has been associated with an increase of gamma-aminobutyric acid (GABA) neurotransmission and, consequently, an excitation/inhibition imbalance associated with autistic-like behavior in both human and animal models. Here, we explored the influence of biological sex in the GABAergic system and behavioral alterations induced by the Nf1 +/− mutation in a murine model. Methods Juvenile male and female Nf1 +/− mice and their wild-type (WT) littermates were used. Hippocampus size was assessed by conventional toluidine blue staining and structural magnetic resonance imaging (MRI). Hippocampal GABA and glutamate levels were determined by magnetic resonance spectroscopy (MRS), which was complemented by western blot for the GABA(A) receptor. Behavioral evaluation of on anxiety, memory, social communication, and repetitive behavior was performed. Results We found that juvenile female Nf1 +/− mice exhibited increased hippocampal GABA levels. Moreover, mutant female displays a more prominent anxious-like behavior together with better memory performance and social behavior. On the other hand, juvenile Nf1 +/− male mice showed increased hippocampal volume and thickness, with a decrease in GABA(A) receptor levels. We observed that mutant males had higher tendency for repetitive behavior. Conclusions Our results suggested a sexually dimorphic impact of Nf1 +/− mutation in hippocampal neurochemistry, and autistic-like behaviors. For the first time, we identified a “camouflaging”-type behavior in females of an animal model of ASD, which masked their autistic traits. Accordingly, like observed in human disorder, in this animal model of ASD, females show larger anxiety levels but better executive functions and production of normative social patterns, together with an imbalance of inhibition/excitation ratio. Contrary, males have more externalizing disorders, such as hyperactivity and repetitive behaviors, with memory deficits. The ability of females to camouflage their autistic traits creates a phenotypic evaluation challenge that mimics the diagnosis difficulty observed in humans. Thus, we propose the study of the Nf1 +/− mouse model to better understand the sexual dimorphisms of ASD phenotypes and to create better diagnostic tools.

O presente artigo estuda a possibilidade de uso de mecanismos de tutela coletiva de direitos para a remoção de barreiras impostas à pessoa com deficiência visual. Para isso, analisa conceitualmente as expressões “pessoa com deficiência” e “deficiência visual”, bem como as chamadas “barreiras”, considerando sua existência e a necessidade de sua remoção como elemento essencial para a inclusão deste grupo. O texto analisa, também, o cabimento e as características da tutela coletiva, concluindo pela adequação de seu uso no caso. Isso porque o artigo destaca que a remoção das barreiras impostas a este grupo revela-se um interesse que ultrapassa o indivíduo, suscitando o cabimento mais eficaz da tutela coletiva. O intuito é, considerando as previsões da LBI, demonstrar como garantir, de forma eficaz, a acessibilidade, quando considerado o enfoque processual. A pesquisa adotou a metodologia de pesquisa jurídico-dogmática por meio de pesquisa bibliográfica.

Common vaccines for infectious diseases have been repurposed as cancer immunotherapies. The intratumoral administration of these repurposed vaccines can induce immune cell infiltration into the treated tumor. Here, we have used an approved trivalent live attenuated measles, mumps, and rubella (MMR) vaccine in our previously developed PeptiENV cancer vaccine platform. The intratumoral administration of this novel MMR-containing PeptiENV cancer vaccine significantly increased both intratumoral as well as systemic tumor-specific T cell responses. In addition, PeptiENV therapy, in combination with immune checkpoint inhibitor therapy, improved tumor growth control and survival as well as increased the number of mice responsive to immune checkpoint inhibitor therapy. Importantly, mice pre-vaccinated with the MMR vaccine responded equally well, if not better, to the PeptiENV therapy, indicating that pre-existing immunity against the MMR vaccine viruses does not compromise the use of this novel cancer vaccine platform.

INTRODUCTION: Vascular graft infections are a rare and severe complication of open aortic surgery. As such, strategies to improve diagnosis and management are paramount. Nevertheless, there is little evidence regarding the factors associated with a higher susceptibility of infection. Moreover, there is little consensus on the best diagnostic workup and most adequate approaches to control and mitigate this surgical complication. We aim to summarize the latest evidence on aortic graft infection through a narrative review. METHODS: We conducted a literature search in a medical database "PubMed" and included studies on vascular graft infection and aortic repair. RESULTS: So far, patient-related comorbidities (such as diabetes, smoking, advanced age and chronic kidney disease), as well as procedure-related factors (location and type of graft, postoperative hyperglycemia, bacteremia, emergency setting), have been reported. Unlike extracavitary infections, aortic graft infections may present with subtle or no clinical manifestations. Aortoenteric fistulas are the exception to the rule, carrying a significantly higher mortality rate. All patients should be thoroughly tested with full blood counts, blood cultures and extensive imaging studies. Adequate antibiotic therapy is one of the pillars of treatment and should not be delayed. The selection of the antimicrobial regimen should be personalized and made in a multidisciplinary team. Surgical strategies are of paramount importance in controlling infection. These can be divided into graft- sparing techniques and graft explantation. The choice of the appropriate approach depends on the surgeon’s experience, the extent of disease and the patient’s general status and comorbidities. CONCLUSION: Aortic graft infections are a rare and severe complication of vascular surgery. A swift diagnosis and management are paramount. Despite significant efforts on how to treat the infection with more adequate antibiotic and surgical therapies, effective preventive measures and a clear definition of predisposing factors remain the main areas for future studies.

Oral insulin delivery could change the life of millions of diabetic patients as an effective, safe, easy-to-use, and affordable alternative to insulin injections, known by an inherently thwarted patient compliance. Here, we designed a multistage nanoparticle (NP) system capable of circumventing the biological barriers that lead to poor drug absorption and bioavailability after oral administration. The nanosystem consists of an insulin-loaded porous silicon NP encapsulated into a pH-responsive lignin matrix, and surface-functionalized with the Fc fragment of immunoglobulin G, which acts as a targeting ligand for the neonatal Fc receptor (FcRn). The developed NPs presented small size (211 ± 1 nm) and narrow size distribution. The NPs remained intact in stomach and intestinal pH conditions, releasing the drug exclusively at pH 7.4, which mimics blood circulation. This formulation showed to be highly cytocompatible, and surface plasmon resonance studies demonstrated that FcRn-targeted NPs present higher capacity to interact and being internalized by the Caco-2 cells, which express FcRn, as demonstrated by Western blot. Ultimately, in vitro permeability studies showed that Fc-functionalized NPs induced an increase in the amount of insulin that permeated across a Caco-2/HT29-MTX co-culture model, showing apparent permeability coefficients (Papp) of 2.37 × 10−6 cm/s, over the 1.66 × 10−6 cm/s observed for their non-functionalized counterparts. Overall, these results demonstrate the potential of these NPs for oral delivery of anti-diabetic drugs.

Resumo Este artigo, elaborado a partir das experiências de três docentes negras, lança um olhar para os processos envolvendo a implementação de políticas afirmativas voltadas para estudantes indígenas. Construímos um percurso que nos leva a refletir desde o Processo Seletivo Especial Indígena, da Universidade Federal do Oeste do Pará, em que duas de nós estivemos diretamente envolvidas, até um debate sobre os reflexos em sala de aula da presença de estudantes vindos das comunidades do oeste do estado do Pará, onde se localiza a Ufopa. Traçamos um panorama analítico sobre os impactos da política de cotas nas vidas de estudantes indígenas e de suas comunidades, bem como nas dinâmicas das universidades desde onde enunciamos, e nas nossas relações, enquanto docentes negras, em sala de aula e na luta política antirracista que se faz dentro da universidade.

Understanding the trophic interactions of introduced predators is key for evidence-based management of biological invasions. This is particularly important in oceanic islands, where predator-prey networks often include numerous endemic and range-restricted species. Geckos are successful island colonizers and in recent years numerous species have established populations in a wide array of oceanic islands. One such species is the Moorish gecko (Tarentola mauritanica), which has colonized multiple islands across the Mediterranean basin, Caribbean and Macaronesia. The species was first reported in Madeira Island in 1993 and over the last 30 years has colonized most of the islands' southern coast and expanded to the nearby island of Porto Santo. Here, we used DNA metabarcoding to provide the first insights into the diet of this successful colonizer in its introduced range. The species' diet was mainly composed of ground-dwelling arthropods belonging to the families Porcellionidae (Isopoda), Julidae (Diplopoda) and Formicidae (Hymenoptera). The diet richness and composition were not affected by neither sex nor size of adult geckos, instead they both change across populations. However, trophic niche-width differed among size classes, with smaller geckos feeding on a wider range of prey. We identified over 160 different Operational Taxonomic Units in the diet of T. mauritanica, with 21.6% of them belonging to introduced invertebrates and 13.6% to native species. Native prey taxa included the endemic Madeira wall lizard (Teira dugesii), the sole native reptile to Madeira. We also detected several agricultural pests and disease vectors in the diet of this exotic predator, and 19 taxa identified as prey had not yet been recorded to Madeira. Of these, several are serious agricultural pests, highlighting how this introduced gecko can be used as a natural sampler, in particular for the early detection of invasive arthropod pests. This study emphasizes the importance of trophic studies for monitoring the impacts of introduced predators in fragile insular systems.

O atual contexto pandémico levou a diversas alterações na rotina dos estudantes de ensino superior, incluindo interrupções de aulas presenciais, implementação do ensino à distância, interrupção ou adiamento de estágios, entre outras, o que pode comprometer a saúde mental. O objetivo deste estudo foi avaliar a perceção da saúde mental em estudantes do ensino superior em tempos de pandemia COVID-19 e os fatores associados. A amostra foi constituída por 1.004 estudantes do ensino superior, com idades compreendidas entre os 18 e 51 anos (20,33±3,35), sendo 725 (72,2%) do sexo feminino. O instrumento incluiu um questionário, elaborado pelos investigadores, dividido em quatro partes: caracterização sócio-demográfica da população, prática de exercício físico, alimentação e saúde mental. Antes do confinamento, 99 (9,9%) estudantes classificaram a sua saúde mental como má e durante o confinamento 423 (42,2%) disseram que a sua saúde mental estava má (p≤0,001). As mulheres apresentaram 1,91 (IC=1,42-2,56; p≤0,001) mais probabilidades de ter uma má saúde mental e os estudantes que não praticaram exercício físico durante o confinamento tiveram 1,56 (IC=1,22-2,04; p≤0,001) mais chances de ter uma saúde mental de má qualidade. A maioria dos estudantes classificou a sua saúde mental como má durante o período de confinamento decorrente da pandemia COVID-19, sendo as mulheres e a ausência de prática de exercício físico os fatores que estiveram mais relacionados com a má saúde mental.

Diabetes is a prevalent global health issue associated with significant morbidity and mortality. Diabetic retinopathy (DR) is a well-known inflammatory, neurovascular complication of diabetes and a leading cause of preventable blindness in developed countries among working-age adults. However, the ocular surface components of diabetic eyes are also at risk of damage due to uncontrolled diabetes, which is often overlooked. Inflammatory changes in the corneas of diabetic patients indicate that inflammation plays a significant role in diabetic complications, much like in DR. The eye’s immune privilege restricts immune and inflammatory responses, and the cornea and retina have a complex network of innate immune cells that maintain immune homeostasis. Nevertheless, low-grade inflammation in diabetes contributes to immune dysregulation. This article aims to provide an overview and discussion of how diabetes affects the ocular immune system’s main components, immune-competent cells, and inflammatory mediators. By understanding these effects, potential interventions and treatments may be developed to improve the ocular health of diabetic patients.

Este estudo propõe uma análise qualitativa das estratégias de comunicação adotadas pela Direção-Geral da Saúde para com a população portuguesa perante a primeira vaga do surto pandémico da COVID-19. Pretende-se, assim, analisar como é que esta entidade, a partir da utilização da rede social Twitter, desenvolveu a divulgação da informação, procurando definir as características que marcam o discurso de cada uma das orientações fornecidas. Para tal, adotou-se o paradigma qualitativo-interpretativo, com base numa análise documental de fontes secundárias, tendo-se adotado a análise temática. No cenário português, observou-se que, relativamente à propagação exponencial do vírus, deu-se a adaptação do fluxo comunicacional, existindo maior recorrência a hashtags, infografias, relatórios de saúde e links externos.

A fim de estudar a viabilidade do direito ao esquecimento no Brasil, este artigo analisa a decisão proferida pelo Supremo Tribunal Federal no julgamento do Tema 786, cujo resultado ensejou entendimentos de que referido direito seria inconstitucional, incompatível com liberdades de expressão e informação. Essa análise revela-se especialmente importante à luz da Sociedade da Informação, em que a preservação de informações atende a interesses de um mercado fundado, justamente, em dados e informações, sobretudo na internet. Para tanto, o trabalho parte da metodologia de pesquisa jurídico-dogmática, presumindo a suficiência da Ciência Jurídica para a compreensão do direito ao esquecimento, ao mesmo tempo em que recorre à metodologia jurídico-sociológica, a qual compreende o Direito num espectro amplo das Ciências Sociais, contextualizando-o à Sociedade Informacional. O estudo conclui que o direito ao esquecimento, per se, não foi declarado inconstitucional; a decisão do STF alcança apenas uma das suas possíveis formas de efetivação.

PurposeTear fluid biomarkers may offer a non-invasive strategy for detecting diabetic patients with increased risk of developing diabetic retinopathy (DR) or increased disease progression, thus helping both improving diagnostic accuracy and understanding the pathophysiology of the disease. Here, we assessed the tear fluid of nondiabetic individuals, diabetic patients with no DR, and diabetic patients with nonproliferative DR (NPDR) or with proliferative DR (PDR) to find putative biomarkers for the diagnosis and staging of DR.MethodsTear fluid samples were collected using Schirmer test strips from a cohort with 12 controls and 54 Type 2 Diabetes (T2D) patients, and then analyzed using mass spectrometry (MS)-based shotgun proteomics and bead-based multiplex assay. Tear fluid-derived small extracellular vesicles (EVs) were analyzed by transmission electron microscopy, Western Blotting, and nano tracking.ResultsProteomics analysis revealed that among the 682 reliably quantified proteins in tear fluid, 42 and 26 were differentially expressed in NPDR and PDR, respectively, comparing to the control group. Data are available via ProteomeXchange with identifier PXD033101. By multicomparison analyses, we also found significant changes in 32 proteins. Gene ontology (GO) annotations showed that most of these proteins are associated with oxidative stress and small EVs. Indeed, we also found that tear fluid is particularly enriched in small EVs. T2D patients with NPDR have higher IL-2/-5/-18, TNF, MMP-2/-3/-9 concentrations than the controls. In the PDR group, IL-5/-18 and MMP-3/-9 concentrations were significantly higher, whereas IL-13 was lower, compared to the controls.ConclusionsOverall, the results show alterations in tear fluid proteins profile in diabetic patients with retinopathy. Promising candidate biomarkers identified need to be validated in a large sample cohort.

Oncolytic viruses (OVs) have been shown to induce anti-cancer immunity and enhance cancer immunotherapies, such as immune checkpoint inhibitor therapies. OV therapies can be further improved by arming OVs with immunostimulatory molecules, including various cytokines or chemokines. Here, we have developed a novel adenovirus encoding two immunostimulatory molecules: cluster of differentiation 40 ligand (CD40L) and tumor necrosis factor receptor superfamily member 4 ligand (OX40L). This novel virus, designated VALO-D102, is designed to activate both innate and adaptive immune responses against tumors. CD40L affects the innate side by licensing antigen-presenting cells to drive CD8+ T cell responses, and OX40L increases clonal expansion and survival of CD8+ T cells and formation of a larger pool of memory T cells. VALO-D102 and its murine surrogate VALO-mD901, expressing murine OX40L and CD40L, were used in our previously developed PeptiCRAd cancer vaccine platform. Intratumoral administration of PeptiCRAd significantly increased tumor-specific T cell responses, reduced tumor growth, and induced systemic anti-cancer immunity in two mouse models of melanoma. In addition, PeptiCRAd therapy, in combination with anti-PD-1 immune checkpoint inhibitor therapy, significantly improved tumor growth control as compared to either monotherapy alone.

Oncolytic adenoviruses have become ideal agents in the path toward treating cancer. Such viruses have been engineered to conditionally replicate in malignant cells in which certain signaling pathways have been disrupted. Other than such oncolytic properties, the viruses need to activate the immune system in order to sustain a long-term response. Therefore, oncolytic adenoviruses have been genetically modified to express various immune-stimulatory agents to achieve this. However, genetically modifying adenoviruses is very time consuming and labor intensive with the current available methods. In this paper, we describe a novel method we have called GAMER-Ad to genetically modify adenovirus genomes within 2 days. Our method entails the replacement of the gp19k gene in the E3 region with any given gene of interest (GOI) using Gibson Assembly avoiding the homologous recombination between the shuttle and the parental plasmid. In this manuscript as proof of concept we constructed and characterized three oncolytic adenoviruses expressing CXCL9, CXCL10, and interleukin-15 (IL-15). We demonstrate that our novel method is fast, reliable, and simple compared to other methods. We anticipate that our method will be used in the future to genetically engineer oncolytic but also other adenoviruses used for gene therapy as well.

O presente estudo possui o objetivo de analisar a usabilidade de programas de DataScience para análise de banco de dados. Para tanto foi utilizado um banco de dados extraído de uma plataforma pública e aplicado em dois programas, sendo Python e Orange, a fim de se obter uma comparação entre ambos. A pesquisa bibliográfica serviu como base para o entendimento dos programas e para embasar os resultados obtidos. O Python exigiu trabalhar o banco de dados, tanto na conversão do arquivo como necessidade de conhecimento e aprendizado de linguagem de programação. No Orange foi utilizado o banco de dados original e sua funcionalidade intuitiva permitiu a obtenção de resultados de forma mais rápida, pois suas ferramentas possuem nomes associados ao que se deseja obter. Com os resultados obtidos foi possível constatar que a utilização do programa Orange se demonstrou mais conveniente para análise do banco de dados manipulado.

Oncolytic Viruses (OVs) work through two main mechanisms of action: the direct lysis of the virus-infected cancer cells and the release of tumor antigens as a result of the viral burst. In this sc.enario, the OVs act as in situ cancer vaccines, since the immunogenicity of the virus is combined with tumor antigens, that direct the specificity of the anti-tumor adaptive immune response. However, this mechanism in some cases fails in eliciting a strong specific T cell response. One way to overcome this problem and enhance the priming efficiency is the production of genetically modified oncolytic viruses encoding one or more tumor antigens. To avoid the long and expensive process related to the engineering of the OVs, we have exploited an approach based on coating OVs (adenovirus and vaccinia virus) with tumor antigens. In this work, oncolytic viruses encoding tumor antigens and tumor antigen decorated adenoviral platform (PeptiCRAd) have been used as cancer vaccines and evaluated both for their prophylactic and therapeutic efficacy. We have first tested the oncolytic vaccines by exploiting the OVA model, moving then to TRP2, a more clinically relevant tumor antigen. Finally, both approaches have been investigated in tumor neo-antigens settings. Interestingly, both genetically modified oncolytic adenovirus and PeptiCRAd elicited T cells-specific anti-tumor responses. However, in vitro cross-representation experiments, showed an advantage of PeptiCRAd as regards the fast presentation of the model epitope SIINFEKL from OVA in an immunogenic rather than tolerogenic fashion. Here two approaches used as cancer oncolytic vaccines have been explored and characterized for their efficacy. Although the generation of specific anti-tumor T cells was elicited in both approaches, PeptiCRAd retains the advantage of being rapidly adaptable by coating the adenovirus with a different set of tumor antigens, which is crucial in personalized cancer vaccines clinical setting.

Besides the isolation and identification of major histocompatibility complex I-restricted peptides from the surface of cancer cells, one of the challenges is eliciting an effective antitumor CD8+ T-cell-mediated response as part of therapeutic cancer vaccine. Therefore, the establishment of a solid pipeline for the downstream selection of clinically relevant peptides and the subsequent creation of therapeutic cancer vaccines are of utmost importance. Indeed, the use of peptides for eliciting specific antitumor adaptive immunity is hindered by two main limitations: the efficient selection of the most optimal candidate peptides and the use of a highly immunogenic platform to combine with the peptides to induce effective tumor-specific adaptive immune responses. Here, we describe for the first time a streamlined pipeline for the generation of personalized cancer vaccines starting from the isolation and selection of the most immunogenic peptide candidates expressed on the tumor cells and ending in the generation of efficient therapeutic oncolytic cancer vaccines. This immunopeptidomics-based pipeline was carefully validated in a murine colon tumor model CT26. Specifically, we used state-of-the-art immunoprecipitation and mass spectrometric methodologies to isolate >8000 peptide targets from the CT26 tumor cell line. The selection of the target candidates was then based on two separate approaches: RNAseq analysis and HEX software. The latter is a tool previously developed by Jacopo, 2020, able to identify tumor antigens similar to pathogen antigens in order to exploit molecular mimicry and tumor pathogen cross-reactive T cells in cancer vaccine development. The generated list of candidates (26 in total) was further tested in a functional characterization assay using interferon-γ enzyme-linked immunospot (ELISpot), reducing the number of candidates to six. These peptides were then tested in our previously described oncolytic cancer vaccine platform PeptiCRAd, a vaccine platform that combines an immunogenic oncolytic adenovirus (OAd) coated with tumor antigen peptides. In our work, PeptiCRAd was successfully used for the treatment of mice bearing CT26, controlling the primary malignant lesion and most importantly a secondary, nontreated, cancer lesion. These results confirmed the feasibility of applying the described pipeline for the selection of peptide candidates and generation of therapeutic oncolytic cancer vaccine, filling a gap in the field of cancer immunotherapy, and paving the way to translate our pipeline into human therapeutic approach.

ABSTRACT BACKGROUND: Poor appetite is common through the aging process and increases the risk of weight loss, protein-energy malnutrition, immunossupression, sarcopenia and frailty. The Simplified Nutritional Appetite Questionnaire (SNAQ) has the aim to monitor appetite and identify older adults at risk of weight loss. OBJECTIVE: To describe the process of translation and cultural adaptation to Brazilian Portuguese of the SNAQ. METHODS: The translation and cultural adaptation was developed in five steps: translation (by three of the authors of the manuscript and assembled by consensus), backtranslation (by an English native speaker), semantic evaluation (by one verontologist and one nutritionist), comprehension of content (by nutrition specialists and by a group of older persons), pre-test and the SNAQ final version development. RESULTS: The SNAQ Portuguese version maintained the original version meaning and referral. To achieve this feature, the process required some modifications to improve the understanding of older persons, such as inclusion of other options to the answers of some questions, rewritten of one question and inclusion of a meal definition. CONCLUSION: SNAQ questionnaire has been successfully translated and adapted to Portuguese. As our next step, we are validating this tool in different clinical settings in Brazil.

Endopleura uchi, is used for the treatment of inflammatory disease and related to the female reproductive tract. The aim of this study was to evaluate the acute toxicity of the Endopleura uchi stem bark hydroethanolic extract (EEu) in zebrafish, emphasizing the histopathological and biochemical parameters, as well as evaluating the in silico pharmacokinetic and toxicological parameters of the phytochemical/pharmacological marker, bergenin, as their metabolites. The animals were orally treated with EEu at a single dose of 75 mg/kg, 500 mg/kg, 1000 mg/kg and 3000 mg/kg. the oral LD50 of the EEu higher to the dose of 3000 mg/kg. Behavioral, biochemical and histopathological changes were dose dependent. In silico pharmacokinetic predictions for bergenin and its metabolites showed moderate absorption in high human intestinal absorption (HIA) and Caco-2 models, reduced plasma protein binding, by low brain tissue binding and no P-glycoprotein (P-Gp) inhibition. Their metabolism is defined by the CYP450 enzyme, in addition to bergenin inhibition of CYP2C9, CYP3A4 and CYP2C19. In the bergenin and its metabolites in silico toxicity test it have been shown to cause carcinogenicity and a greater involvement of the bergenin with the CYP enzymes in the I and II hepatic and renal metabolism’s phases was observed. It is possible to suggest that the histopathological damages are involved with the interaction of this major compound and its metabolites at the level of the cellular-biochemical mechanisms which involve the absorption, metabolization and excretion of these possible prodrug and drug.

Cancer therapy using oncolytic virus has shown pre-clinical and clinical efficacy. Here, the authors report ExtraCRAd, an oncolytic virus cloaked with tumour cell membrane and report its therapeutic effects in vitro and in vivo in multiple mouse tumour models.

Resumo: Este artigo, escrito por três mulheres, professoras, busca refletir sobre o ensino de História em sua dimensão corporal. Quando uma professora ensina história para um grupo de estudantes, aqueles espaçostempos são ocupados pelo corpo da professora, pelos corpos de todas as estudantes e pelos corpos de todas que fazem a história e habitam as narrativas históricas daqueles povos. São muitos corpos que por vezes são entendidos de forma metonímica, como apenas mentes que pensam. E pensam outros corpos tomados apenas como números em estatísticas na narrativa histórica. Buscando romper com esta separação entre mente e corpo, refletindo com estes corpos, seus processos identitários e suas formas de se expressar, este texto traz narrativas sobre os corpos docentes, estudantis e das personagens históricas de nossas aulas. A partir de três narrativas experimentadas pelas professoras/pesquisadoras em suas salas de aula de história, disputamos novos sentidos para o ensino de história. Apostamos em um ensino de histórias no plural, produzido e experimentado por diferentes corpos.

Background Despite the success of immune checkpoint inhibitors against PD-L1 in the clinic, only a fraction of patients benefit from such therapy. A theoretical strategy to increase efficacy would be to arm such antibodies with Fc-mediated effector mechanisms. However, these effector mechanisms are inhibited or reduced due to toxicity issues since PD-L1 is not confined to the tumor and also expressed on healthy cells. To increase efficacy while minimizing toxicity, we designed an oncolytic adenovirus that secretes a cross-hybrid Fc-fusion peptide against PD-L1 able to elicit effector mechanisms of an IgG1 and also IgA1 consequently activating neutrophils, a population neglected by IgG1, in order to combine multiple effector mechanisms.Methods The cross-hybrid Fc-fusion peptide comprises of an Fc with the constant domains of an IgA1 and IgG1 which is connected to a PD-1 ectodomain via a GGGS linker and was cloned into an oncolytic adenovirus. We demonstrated that the oncolytic adenovirus was able to secrete the cross-hybrid Fc-fusion peptide able to bind to PD-L1 and activate multiple immune components enhancing tumor cytotoxicity in various cancer cell lines, in vivo and ex vivo renal-cell carcinoma patient-derived organoids.Results Using various techniques to measure cytotoxicity, the cross-hybrid Fc-fusion peptide expressed by the oncolytic adenovirus was shown to activate Fc-effector mechanisms of an IgA1 (neutrophil activation) as well as of an IgG1 (natural killer and complement activation). The activation of multiple effector mechanism simultaneously led to significantly increased tumor killing compared with FDA-approved PD-L1 checkpoint inhibitor (Atezolizumab), IgG1-PDL1 and IgA-PDL1 in various in vitro cell lines, in vivo models and ex vivo renal cell carcinoma organoids. Moreover, in vivo data demonstrated that Ad-Cab did not require CD8+ T cells, unlike conventional checkpoint inhibitors, since it was able to activate other effector populations.Conclusion Arming PD-L1 checkpoint inhibitors with Fc-effector mechanisms of both an IgA1 and an IgG1 can increase efficacy while maintaining safety by limiting expression to the tumor using oncolytic adenovirus. The increase in tumor killing is mostly attributed to the activation of multiple effector populations rather than activating a single effector population leading to significantly higher tumor killing.

Background Intratumoral BCG therapy, one of the earliest immunotherapies, can lead to infiltration of immune cells into a treated tumor. However, an increase in the number of BCG-induced tumor-specific T cells in the tumor microenvironment could lead to enhanced therapeutic effects.Methods Here, we have developed a novel cancer vaccine platform based on BCG that can broaden BCG-induced immune responses to include tumor antigens. By physically attaching tumor-specific peptides onto the mycobacterial outer membrane, we were able to induce strong systemic and intratumoral T cell-specific immune responses toward the attached tumor antigens. These therapeutic peptides can be efficiently attached to the mycobacterial outer membrane using a poly-lysine sequence N-terminally fused to the tumor-specific peptides.Results Using two mouse models of melanoma and a mouse model of colorectal cancer, we observed that the antitumor immune responses of BCG could be improved by coating the BCG with tumor-specific peptides. In addition, by combining this novel cancer vaccine platform with anti-programmed death 1 (anti-PD-1) immune checkpoint inhibitor (ICI) therapy, the number of responders to anti-PD-1 immunotherapy was markedly increased.Conclusions This study shows that intratumoral BCG immunotherapy can be improved by coating the bacteria with modified tumor-specific peptides. In addition, this improved BCG immunotherapy can be combined with ICI therapy to obtain enhanced tumor growth control. These results warrant clinical testing of this novel cancer vaccine platform.

Este estudo teve como objetivo principal a investigação da produção arquitetônica na Belém dos anos 1940, no período em que as primeiras construções modernistas surgiam em forma da verticalização da Avenida 15 de agosto e a linguagem neocolonial, inspirada do conceito de identidade nacional, despontava, ambas convivendo com as edificações ecléticas. Para isto, realizou-se esta análise à luz da produção do Arquiteto português David Ferreira de Oliveira Lopes (1911-1987), premiado profissional atuante nesta década na cidade, de forma a notar os trânsitos culturais transatlânticos expressos na Arquitetura. Foram adotados modelos para que estas arquiteturas pudessem ser compreendidas sob a perspectiva da modernização da cidade e das influências entre Portugal e Brasil. Perspectiva essa que norteava a Arquitetura não somente como o desenho do espaço, do habitar e do frequentar, mas como uma das Belas Artes, não à toa eram realizadas premiações aos melhores projetos de arquitetura da cidade, nos chamados Salões Oficiais de Belas Artes do Estado do Pará, tendo sido o próprio David Lopes homenageado com diversas destas premiações. Trazer à luz as obras e projetos deste arquiteto permite entender o longo processo em que o tradicionalismo se conjugou aos anseios de modernização na produção da metrópole da Amazônia durante o século XX.

Neste artigo investigamos a distinção massivo-contável na língua Rikbaktsa (Macro-Jê). Partimos do questionário de Lima & Rothstein (2020), o qual procuramos responder levantando os dados em um acervo inédito do qual já dispúnhamos, na pouca literatura sobre a língua (Boswood 1971, 1978; SIL 2007; Silva 2011), e em uma elicitação de dados com um falante nativo. A análise mostrou que há morfologia de plural e que ela não se combina com nomes de massa. Nomes que denotam átomos estáveis se combinam diretamente com numerais. Para serem contados, nomes de substância exigem a presença de sintagmas de medidas. Esses são indícios de que a língua em questão pode ser caracterizada como uma língua de número marcado (Chierchia 2010, 2015), mesmo que haja alguns nomes de massa que podem ser contados diretamente. Em nenhuma das quinze línguas apresentadas em Lima & Rothstein (2020) há relato de um morfema especializado para marcar massa. Chacon (2012) afirma que o Kubeo (Tukano-Oriental) exibe um demonstrativo de massa. Esse é também o caso em Rikbaktsa, mas nesta língua o demonstrativo na ‘isto’, que seleciona nomes massivos, se espalha pela gramática: aparece com marca de terceira pessoa no pronome pessoal a-na, com proformas interrogativas e em construção atributiva. Este estudo contribui, portanto, para uma melhor compreensão da distinção massivo-contável através das línguas e da gramática desta língua minoritária.

Nos últimos 71 anos foram realizadas cerca de 71 operações de paz sob o comando das Nações Unidas. Esse expressivo número evidencia que as operações são entendidas como uma importante ferramenta para a manutenção da paz e segurança internacional, conforme o proposto na Carta das Nações. A maior parte das operações de paz aconteceram após a Guerra Fria e por isso, o presente trabalho busca compreender como se dá a participação dos países da América do Sul nessas operações, uma vez que a contribuição de alguns desses países acontece desde os primórdios de tais missões. Para isso, o artigo primeiramente explica como era a participação dos países sulamericanos durante a Guerra Fria, enquanto o mundo encontrava-se dividido entre duas ideologias. Depois busca mostrar a evolução das operações de paz quando a Guerra acabou. Em seguida, o trabalho mostra como os países sulamericanos passaram a engajar em operações de paz no pós-Guerra Fria e qual o propósito dessas participações.

The present study was conducted to investigate the risk factors for post-mortem findings and causes of sow mortality. A post-mortem examination and microbiological investigation were conducted on 123 sows from a breeding herd with 15,000 dams. The mortality of spontaneous death in sows occurred mostly in the peripartum period (53%; p < 0.05). The spontaneous deaths were associated with heart failures, hemorrhagic and perforating gastric ulcers, and liver torsion, while in the euthanized sows, the post-mortem findings were associated with locomotor disorders. A higher body condition score (BCS ≥ 3.5) increased (p < 0.05) heart failure on the post-mortem examination. The excessive use of manual obstetric interventions increased sow deaths resulting from cervix/uterus ruptures and increased the odds of death (p < 0.05) due to metritis. Sow mortality had a multifactorial etiology. Infections were polymicrobial. The main microbial agents identified from a septic lesion in locomotor, genitourinary, and respiratory systems were Trueperella pyogenes, Escherichia coli, and Actinobacillus pleuropneumoniae, respectively. In conclusion, sow mortality involved multiple risk factors and several bacterial agents. These results indicate that better management practices can reduce sow mortality in swine production and increase sow welfare.

Messenger RNA (mRNA) and microRNA (miRNA)-based therapeutics have become attractive alternatives to DNA-based therapeutics due to recent advances in manufacture, scalability and cost. Also, RNA-based therapeutics are considered safe since there are no risk of inducing genomic changes as well as the potential adverse effects would be only temporary due to the transient nature of RNA-based therapeutics. However, efficient in vivo delivery of RNA-based therapeutics remains a challenge. We have developed a delivery platform for RNA-based therapeutics by exploiting the physicochemical properties of enveloped viruses. By physically attaching cationic liposome/RNA complexes onto the viral envelope of vaccinia virus, we were able to deliver mRNA, self-replicating RNA as well as miRNA inside target cells. Also, we showed that this platform, called viRNA platform, can efficiently deliver functional miRNA mimics into B16.OVA tumour in vivo.

This paper discusses in five sections the permissibility of a hypothetical conflict for the Guarani Aquifer resources. First, we introduce the problematic of cross-border conflict related to the usage of groundwater in South America, highlighting the importance of a conscious reflection on the morality behind resource-related conflicts. Next, we move on to a brief explanation of the modern just war theory, emphasizing its focus on just cause as a condition a conflict must meet in order to be justified from an ius ad bellum perspective. Then, we present Conway Waddington’s criticism to just war theory, together with a formulation of our hypothetical problem. In the fourth section of the paper, we defend pragmatism as a methodological approach to contemporary resource-related conflicts, for it can replace a just war theory that has prescriptive voids, and we call the reader’s attention to the fact that international law shows much influence from pragmatic thinking, setting principles that must be followed but showing concern for its historical adaptability from time to time. Finally, we draw conclusions from our analysis, and they should help the reader to maintain a positive outlook towards the theoretical and practical challenges posed to international law by water scarcity. All along, our goal is to stimulate the reader to think critically and develop his or her own assessment of those pressing moral and legal issues, hoping to raise awareness of the much problematic relation between lack of water security and imminence of armed conflict. Although we know a war is unlikely to happen in South America, we should discuss the moral, legal and humanitarian implications of water policies around the world, and instigate scientists to think of solutions for resource scarcity. This is our attempt to kick-start debate around each one’s personal responsibility for demanding fair and equal water distribution.

Methylmercury (MeHg) toxicity is a major environmental concern. In the aquatic reservoir, MeHg bioaccumulates along the food chain until it is consumed by riverine populations. There has been much interest in the neurotoxicity of MeHg due to recent environmental disasters. Studies have also addressed the implications of long-term MeHg exposure for humans. The central nervous system is particularly susceptible to the deleterious effects of MeHg, as evidenced by clinical symptoms and histopathological changes in poisoned humans. In vitro and in vivo studies have been crucial in deciphering the molecular mechanisms underlying MeHg-induced neurotoxicity. A collection of cellular and molecular alterations including cytokine release, oxidative stress, mitochondrial dysfunction, Ca2+ and glutamate dyshomeostasis, and cell death mechanisms are important consequences of brain cells exposure to MeHg. The purpose of this review is to organize an overview of the mercury cycle and MeHg poisoning events and to summarize data from cellular, animal, and human studies focusing on MeHg effects in neurons and glial cells. This review proposes an up-to-date compendium that will serve as a starting point for further studies and a consultation reference of published studies.

The Brazilian Constitution of 1988 guaranteed everyone a healthy and ecologically balanced environment. And for this it is necessary that any human activity, especially industrial, that interfere in the environment be controlled. Solid waste configures itself as a major component of the environmental impacts produced by industrial activities. For this reason, they need to be monitored and controlled throughout its production. The oil industry is classified as highly polluting, and one reason for this categorization is the generation of solid waste. This study aims at conducting an analysis of the National Solid Waste Policy in Brazil, published in 2010, and the adequacy of the oil industry as to its content. The methodology used was a literature review for a survey of the environmental legal framework applicable in this subject, and afterward environmental laws applied to the case were compared and analyzed for consistency and legality; finally, from the data explained in IBAMA’s Technical Note regarding the solid waste produced by the oil industry, omissions and inconsistencies were pointed to the current model of sustainable development adopted by the CF/88. Thus, we can see that an alignment is required within Brazilian standards, as well as meeting the requirements of the oil industry on the solid waste produced in this industrial activity.

Diabetic retinopathy (DR) is a complex, progressive, and heterogenous retinal degenerative disease associated with diabetes duration. It is characterized by glial, neural, and microvascular dysfunction, being the blood-retinal barrier (BRB) breakdown a hallmark of the early stages. In advanced stages, there is formation of new blood vessels, which are fragile and prone to leaking. This disease, if left untreated, may result in severe vision loss and eventually legal blindness. Although there are some available treatment options for DR, most of them are targeted to the advanced stages of the disease, have some adverse effects, and many patients do not adequately respond to the treatment, which demands further research. Oxidative stress and low-grade inflammation are closely associated processes that play a critical role in the development of DR. Retinal cells communicate with each other or with another one, using cell junctions, adhesion contacts, and secreted soluble factors that can act in neighboring or long-distance cells. Another mechanism of cell communication is via secreted extracellular vesicles (EVs), through exchange of material. Here, we review the current knowledge on deregulation of cell-to-cell communication through EVs, discussing the changes in miRNA expression profiling in body fluids and their role in the development of DR. Thereafter, current and promising therapeutic agents for preventing the progression of DR will be discussed.

A presente investigação tem como objetivo atenuar as perdas e minimizar custos no setor de hortifrúti de um supermercado varejista aplicando as ferramentas da qualidade. Realizou-se um levantamento das perdas do setor, aplicando as ferramentas da qualidade para diagnóstico da origem do problema e tomada de decisão, e por fim foi realizada uma análise dos dados com o intuito de propor a metodologia adequada para a solução do problema. A pesquisa foi de natureza quali-quantitativa com caráter exploratório tendo como método de investigação o estudo de caso. Como instrumento para a coleta de dados foi utilizada a pesquisa bibliográfica, pesquisa documental e observação. Os resultados obtidos mostraram que os fatores que provocam ou auxiliam o aumento das perdas são o excesso de umidade relativa do ar, transporte, manuseio incorreto ou excessivo e acondicionamento não adequado.

Em 2012, no âmbito do projeto de Pesquisa e Extensão Mapeamento das Casas e Terreiros de Religiões de Matriz Afro-brasileira na Cidade de Santarém/Pará, reunimos um grupo de 20 estudantes da Universidade Federal do Oeste do Pará (UFOPA), alguns professores, e Afro-religiosos, no terreiro de Mina Santa Bárbara, espaço sagrado mantido por Pai Edvanei de Oyá. Durante dois dias estivemos aprendendo cânticos e toques presentes no repertório Afro-religioso de Terreiros, que são parte constitutiva da paisagem sonora, arquitetônica, Afro-diaspórica e histórica de Santarém. Este momento de fusão Terreiro-Academia foi a ponta de lança para uma série ininterrupta de ações, diálogos, afetos, ajuda e solidariedade que ainda nos une. A primeira parte deste artigo persegue uma questão colocada por Gomes (2010) sobre o que está colocado para o intelectual negro politicamente posicionado que atua nos espaços acadêmicos. Seguindo a proposta inaugurada com a Oficina de Toques e Cantigas do Repertório dos Terreiros, em 2016 realizamos, dentro da UFOPA, o minicurso Antropologia Feminista Negra: Experiência Vivida, Ativismo Político, Interseccionalidade e Sexualidades, no qual tivemos a honra de assistir uma aula ministrada pela Iyalorixá Ozanélia Santos. Ali, além de partilhar nossas experiências enquanto mulheres negras, indígenas, quilombolas, afro-religiosas, educadoras, estudantes, partilhamos do axé, da escuta e do diálogo com a Iyalorixá. Nas próximas páginas vamos discutir alguns aspectos do desafio de ensinar e aprender a partir de epistemologias que nos autorizam falar, que nos fazem visíveis, nos mantêm seguros, e nos permitem ter prazer fazendo o que gostamos e escolhemos fazer. Estas são as epistemologias dos Terreiros, ou os modos de saber e de viver Negro-brasileiro.

OBJECTIVE: To determine the underdiagnosis rate in new COPD cases at the end of a nine-year follow-up period-in the study designated "Projeto Latino-Americano de Investigação em Obstrução Pulmonar" (PLATINO, Latin-American Pulmonary Obstruction Investigation Project)-and compare that with the underdiagnosis rate during the initial phase of the study, as well as to identify the clinical features exhibited by the subjects who were not diagnosed until the end of the follow-up phase. METHODS: The study population comprised the 1,000 residents of the city of São Paulo, Brazil, who took part in the PLATINO study. Of those, 613 participated in the follow-up phase, during which the subjects were assessed with the same instruments and equipment employed in the initial phase of the study. We used the chi-square test or the independent sample t-test to analyze the underdiagnosis rate and to identify the characteristics of the subjects who were not diagnosed until the end of the follow-up phase. RESULTS: The underdiagnosis rate for new COPD cases at the end of the nine-year follow-up period was 70.0%. The underdiagnosis rate during the follow-up phase was 17.5% lower than that reported for the initial phase of the study. The subjects who were not diagnosed until the end of the follow-up phase presented with fewer respiratory symptoms, better pulmonary function, and less severe disease than did those previously diagnosed with COPD. CONCLUSIONS: The underdiagnosis rate for new COPD cases was lower in the follow-up phase of the study than in the initial phase. The subjects who were not diagnosed until the end of the follow-up phase of the PLATINO study presented with the same clinical profile as did those who were not diagnosed in the initial phase. These findings underscore the need for spirometry in order to confirm the diagnosis of COPD and provide early intervention.

OBJECTIVE: To validate and develop an immunonephelometric assay for the determination of alpha-1 antitrypsin (AAT) levels in dried blood spots from COPD patients in Brazil. METHODS: We determined AAT levels in serum samples and dried blood spots from 192 COPD patients. For the preparation of dried blood spots, a disk (diameter, 6 mm) was placed into a tube, eluted with 200 µL of PBS, and stored overnight at 4ºC. All of the samples were analyzed by immunonephelometry in duplicate. We used the bootstrap resampling method in order to determine a cut-off point for AAT levels in dried blood spots. RESULTS: The correlation coefficient between the AAT levels in serum samples and those in dried blood spots was r = 0.45. For dried blood spots, the cut-off value was 2.02 mg/dL (97% CI: 1.45-2.64 mg/dL), with a sensitivity, specificity, positive predictive value, and negative predictive value of 100%, 95.7%, 27.2%, and 100%, respectively. CONCLUSIONS: This method for the determination of AAT levels in dried blood spots appears to be a reliable screening tool for patients with AAT deficiency.

Boar semen cryopreservation remains a challenge due to the extension of cold shock damage. Thus, many alternatives have emerged to improve the quality of frozen-thawed boar sperm. Although the use of seminal plasma arising from boar sperm-rich fraction (SP-SRF) has shown good efficacy; however, the majority of actual sperm evaluation techniques include a single or dual sperm parameter analysis, which overrates the real sperm viability. Within this context, this work was performed to introduce a sperm flow cytometry fourfold stain technique for simultaneous evaluation of plasma and acrosomal membrane integrity and mitochondrial membrane potential. We then used the sperm flow cytometry fourfold stain technique to study the effect of SP-SRF on frozen-thawed boar sperm and further evaluated the effect of this treatment on sperm movement, tyrosine phosphorylation and fertility rate (FR). The sperm fourfold stain technique is accurate (R2 = 0.9356, p > 0.01) for simultaneous evaluation of plasma and acrosomal membrane integrity and mitochondrial membrane potential (IPIAH cells). Centrifugation pre-cryopreservation was not deleterious (p > 0.05) for any analyzed variables. Addition of SP-SRF after cryopreservation was able to improve total and progressive motility (p < 0.05) when boar semen was cryopreserved without SP-SRF; however, it was not able to decrease tyrosine phosphorylation (p > 0.05) or improve IPIAH cells (p > 0.05). FR was not (p > 0.05) statistically increased by the addition of seminal plasma, though females inseminated with frozen-thawed boar semen plus SP-SRF did perform better than those inseminated with sperm lacking seminal plasma. Thus, we conclude that sperm fourfold stain can be used to simultaneously evaluate plasma and acrosomal membrane integrity and mitochondrial membrane potential, and the addition of SP-SRF at thawed boar semen cryopreserved in absence of SP-SRF improve its total and progressive motility.

FUNDAMENTO: O tabagismo altera a função autonômica. OBJETIVO: Investigar os efeitos agudos do tabagismo sobre a modulação autonômica e a recuperação dos índices de variabilidade de frequência cardíaca (VFC) pós-fumo, por meio do plot de Poincaré e índices lineares. MÉTODOS: Foram avaliados 25 fumantes jovens, os quais tiveram a frequência cardíaca analisada, batimento a batimento, na posição sentada, após 8 horas de abstinência, por 30 minutos em repouso, 20 minutos durante o fumo e 30 minutos pós-fumo. Análise de variância para medidas repetidas, seguido do teste de Tukey, ou teste de Friedman seguido do teste de Dunn foram aplicados dependendo da normalidade dos dados, com p < 0,05. RESULTADOS: Durante o fumo, houve redução dos índices SD1 (23,4 ± 9,2 vs 13,8 ± 4,8), razão SD1/SD2 (0,31 ± 0,08 vs 0,2 ± 0,04), RMSSD (32,7 ± 13 vs 19,1 ± 6,8), SDNN (47,6 ± 14,8 vs 35,5 ± 8,4), HFnu (32,5 ± 11,6 vs 19 ± 8,1) e do intervalo RR (816,8 ± 89 vs 696,5 ± 76,3) em relação ao repouso, enquanto que aumentos do índice LFnu (67,5 ± 11,6 vs 81 ± 8,1) e da razão LF/HF (2,6 ± 1,7 vs 5,4 ± 3,1) foram observados. A análise visual do plot mostrou menor dispersão dos intervalos RR durante o fumo. Com exceção da razão SD1/SD2, os demais índices apresentaram recuperação dos valores, 30 minutos após o tabagismo. CONCLUSÃO: O tabagismo produziu agudamente modificações no controle autonômico, caracterizadas por ativação simpática e retirada vagal, com recuperação 30 minutos após o fumo.FUNDAMENTO: El tabaquismo altera la función autonómica. OBJETIVO: Investigar los efectos agudos del tabaquismo sobre la modulación autonómica y la recuperación de los índices de variabilidad de frecuencia cardíaca (VFC) post tabaco, por medio del plot de Poincaré e índices lineales. MÉTODOS: Fueron evaluados 25 fumantes jóvenes, los cuales tuvieron la frecuencia cardíaca analizada, latido a latido, en la posición sentada, después de 8 horas de abstinencia, por 30 minutos en reposo, 20 minutos durante el tabaco y 30 minutos post tabaco. Análisis de varianza para medidas repetidas, seguido del test de Tukey, o test de Friedman seguido del test de Dunn fueron aplicados dependiendo de la normalidad de los datos, con p < 0,05. RESULTADOS:: Durante el tabaco, hubo reducción de los índices SD1 (23,4 ± 9,2 vs 13,8 ± 4,8), razón SD1/SD2 (0,31 ± 0,08 vs 0,2 ± 0,04), RMSSD (32,7 ± 13 vs 19,1 ± 6,8), SDNN (47,6 ± 14,8 vs 35,5 ± 8,4), HFnu (32,5 ± 11,6 vs 19 ± 8,1) y del intervalo RR (816,8 ± 89 vs 696,5 ± 76,3) en relación al reposo, mientras que aumentos del índice LFnu (67,5 ± 11,6 vs 81 ± 8,1) y de la razón LF/HF (2,6 ± 1,7 vs 5,4 ± 3,1) fueron observados. El análisis visual del plot mostró menor dispersión de los intervalos RR durante el tabaco. Con excepción de la razón SD1/SD2, los demás índices presentaron recuperación de los valores, 30 minutos después del tabaquismo. CONCLUSIÓN: El tabaquismo produjo agudamente modificaciones en el control autonómico, caracterizadas por activación simpática y retirada vagal, con recuperación 30 minutos después del tabaco.BACKGROUND: Smoking affects the autonomic function. OBJECTIVE:To investigate the acute effects of smoking on the autonomic modulation and the post-smoking recovery of the heart rate variability (HRV) index through Poincare plot and linear indices. METHODS: A total of 25 young smokers underwent beat-to-beat analysis of heart rate in the sitting position after 8 hours of tobacco abstinence, for 30 minutes at rest, 20 minutes while smoking and 30 minutes after smoking. The analysis of variance for repeated measures, followed by Tukey's test or Friedman's test followed by Dunn's test, were applied depending on the normality of data, with p < 0.05. RESULTS: While smoking, there was a decrease in the indices: SD1 (23.4 ± 9.2 vs 13.8 ± 4.8), SD1/SD2 ratio (0.31 ± 0.08 vs 0.2 ± 0.04), RMSSD (32,7 ± 13 vs 19.1 ± 6.8), SDNN (47.6 ± 14.8 vs 35.5 ± 8.4), HFnu (32,5 ± 11.6 vs 19 ± 8.1) and the RR interval (816.8 ± 89 vs 696.5 ± 76.3) in relation to the rest period, whereas increases in the LFnu index (67.5 ± 11.6 vs 81 ± 8.1) and the LF/HF ratio (2.6 ± 1.7 vs 5.4 ± 3.1) were observed. The visual analysis of the plot showed a lower dispersion of RR intervals while smoking. Except for the SD1/SD2 ratio, the other indices presented recovery of values 30 minutes after smoking. CONCLUSION: Smoking resulted in acute modifications of the autonomic control, characterized by sympathetic activation and vagal withdrawal, presenting recovery 30 minutes after smoking.

To determine parameters associated with the evolution of sepsis, a five-year retrospective study was conducted in a university hospital. One hundred and four consecutive sepsis patients were evaluated, of whom 55.8% were men. The mortality was 68.3% and was associated with older age (pPara determinar parâmetros associados à evolução da sepse, foi realizado estudo retrospectivo de 5 anos em um hospital universitário. Foram avaliados 104 pacientes consecutivos com sepse, sendo 55,8% homens. A mortalidade foi de 68,3%, associada à idade elevada (p

PURPOSE: To investigate the effects of different conditions used during cardiopulmonary bypass (CPB) surgery on accompanying production of cytokine and nitric oxide (NO). METHODS: Patients undergoing CPB for the first time were prospectively enrolled and divided into two groups according to CPB parameters performed: i) normothermia (36.5-37°C) with blood cardioplegia (NB group, n=10) and ii) hypothermia (29-31°C) with crystalloid cardioplegia (HC group, n=10). Plasma samples obtained following intubation (baseline), during (5 and 30 min) and after (4 and 24 h) CPB were assayed for cytokines (ELISA) and NO metabolites (Griess reaction). RESULTS: Peak concentrations of interleukin (IL)-6 and IL-8 were reached at 4 h post CPB in both groups, but in the HC group those levels increased earlier and persisted for longer (24 h) compared to baseline (P < 0.05). IL-10 levels also increased at 4 h compared to baseline, but only significantly so in the HC group. NO metabolites were reduced in HC group at all time points compared to baseline (P < 0.05), while no significant differences were detected in the NB group. CONCLUSION: The association between increased systemic levels of cytokines and reduced NO production in the HC group suggests that different myocardial protection and/or perfusion temperature used during CPB may contribute to the extent of inflammatory response.OBJETIVO: Investigar a hipótese de que diferentes procedimentos durante o bypass cardiopulmonary (BCP) causa diferentes níveis de citocinas (IL) e óxido nítrico (NO). MÉTODOS: Pacientes submetidas a BCP foram prospectivamente estudadas de acordo com bypass realizado sobre normotermia (36.5-37°C) com cardioplegia sanguínea (NB group, n=10) or hipotermia (29-31°C) com cardioplegia cristalóide (HC group, n=10). Amostras de Plasma foram obtidas após a intubação (linha de base), durante (5, 30 min) e após (4, 24 h) o BCP. Os ensaios foram realizados através de ELISA (IL) e metabólitos do NO (reação de Griess). RESULTADOS: Os picos de concentrações de IL-6 and IL-8 estavam aumentados em 4 h pós BCP em ambos os grupos, mas no grupo HC estes níveis aumentaram precocemente e persistiram aumentadas por 24 h, comparado a linha de base (P

Sepsis is a systemic inflammatory response commonly caused by bacterial infection. We demonstrated that the outcome of sepsis induced by cecal ligation and puncture (CLP) correlates with the severity of the neutrophil migration failure towards infectious focus. Failure appears to be due to a decrease in the rolling and adhesion of neutrophil to endothelium cells. It seems that neutrophil migration impairment is mediated by the circulating inflammatory cytokines, such as TNF-alpha and IL-8, which induce the nitric oxide (NO) production systemically. It is supported by the fact that intravenous administration of these cytokines reduces the neutrophil migration induced by different inflammatory stimuli, and in severe sepsis the circulating concentrations of the cytokines and chemokines are significantly increased. Moreover, the neutrophil migration failure and the reduction in the rolling/adhesion were not observed in iNOS-/- mice and, aminoguanidine prevented this event. We also demonstrated that the failure of neutrophil migration is a Toll-4 receptor (TLR4) dependent mechanism, since it was not observed in TLR4 deficient mice. Furthermore, it was also observed that circulating neutrophils obtained from septic patients present failure of neutrophil chemotaxis toward fMLP, IL-8, and LTB4 and an increased in sera concentrations of NO3 and cytokines. In conclusion, we demonstrated that, in sepsis, failure of neutrophil migration is critical for the outcome and that NO is involved in the process.