Role of CD4+ T-cells for regulating splenic myelopoiesis and monocyte differentiation after experimental myocardial infarction.

Myocardial infarction (MI) induces the generation of proinflammatory Ly6Chigh monocytes in the spleen and the recruitment of these cells to the myocardium. CD4+ Foxp3+ CD25+ T-cells (Tregs) promote the healing process after myocardial infarction by engendering a pro-healing differentiation state in myocardial monocyte-derived macrophages. We aimed to study the effects of CD4+ T-cells on splenic myelopoiesis and monocyte differentiation. We instigated MI in mice and found that MI-induced splenic myelopoiesis is abrogated in CD4+ T-cell deficient animals. Conventional CD4+ T-cells promoted myelopoiesis in vitro by cell-cell-contact and paracrine mechanisms, including interferon-gamma (IFN-γ) signalling. Depletion of regulatory T-cells enhanced myelopoiesis in vivo, as evidenced by increases in progenitor cell numbers and proliferative activity in the spleen 5 days after MI. The frequency of CD4+ T-cells-producing factors that promote myelopoiesis increased within the spleen of Treg-depleted mice. Moreover, depletion of Tregs caused a proinflammatory bias in splenic Ly6Chigh monocytes, which showed predominantly upregulated expression of IFN-γ responsive genes after MI. Our results indicate that conventional CD4+ T-cells promote and Tregs attenuate splenic myelopoiesis and proinflammatory differentiation of monocytes.

Characterizing the immune response to myocardial infarction in pigs.

Though myocardial infarction (MI) in pigs is a well-established translational large animal model, it has not yet been widely used for immunotherapy studies, and a comprehensive description of the immune response to MI in this species is lacking. We induced MI in Landrace pigs by balloon occlusion of the left anterior descending artery over 90 min. Within 14 days, the necrotic myocardium was progressively replaced by scar tissue with involvement of myofibroblasts. We characterized the immune response in the heart ex vivo by (immuno)histology, flow cytometry, and RNA sequencing of myocardial tissue on days 3, 7, and 14 after MI. Besides a clear predominance of myeloid cells among heart-infiltrating leukocytes, we detected activated T cells and an increasing proportion of CD4+ Foxp3+ regulatory T cells (Treg), especially in the infarct core-findings that closely mirror what has been observed in mice and humans after MI. Transcriptome data indicated inflammatory activity that was persistent but markedly changing in character over time and linked to extracellular matrix biology. Analysis of lymphocytes in heart-draining lymph nodes revealed significantly higher proliferation rates of T helper cell subsets, including Treg on day 7 after MI, compared to sham controls. Elevated frequencies of myeloid progenitors in the spleen suggest that it might be a site of emergency myelopoiesis after MI in pigs, as previously shown in mice. We thus provide a first description of the immune response to MI in pigs, and our results can aid future research using the species for preclinical immunotherapy studies.

Fatores associados ao autocuidado praticado por pessoas com estomias de eliminação / Factors associated with self-care practiced by people with intestinal elimination stomas / Factores asociados al autocuidado practicado por personas con estomas de eliminación

Objetivo: avaliar os fatores associados ao autocuidado praticado por pessoas com estomias de eliminação. Método: estudo transversal analítico, baseado nas respostas de 153 pessoas com estomia de eliminação, atendidas em um ambulatório de referência no cuidado de pessoas com estomas, no município de Teresina. A coleta de dados ocorreu por meio de um formulário semiestruturado com dados demográficos e acerca do autocuidado. Os dados foram analisados mediante estatística descritiva e inferencial. O teste Qui-quadrado de Pearson foi utilizado na análise inferencial. Resultados: o esvaziamento da bolsa, limpeza do estoma, secagem da pele periestoma, descolamento da placa, medição do estoma, realização do molde, adaptação, autoestima e isolamento social apresentaram associação significativa com o autocuidado (p<0,05). Conclusão: evidenciou-se que os fatores associados ao autocuidado foram os cuidados com o estoma e com o equipamento coletor, além dos impactos na autoestima e na vida social.

Objective: to evaluate the factors associated with self-care practiced by people with elimination stomas. Method: analytical cross-sectional study, based on the responses of 153 people with elimination stoma, treated at a reference outpatient clinic for the care of people with stoma, in the city of Teresina. Data collection took place through a semi-structured form with demographic data and about self-care. Analyzes were performed using inferential statistics, using the chi-square test. Results: Emptying the pouch, cleaning the stoma, drying the peristomal skin, detaching the plaque, measuring the stoma, making the mold, fitting, , self-esteem and social isolation were significantly associated with self-care (p<0.05). Conclusion: it was evident that the factors associated with self-care were care for the stoma and the collection equipment, in addition to the impacts on self-esteem and social life.

Objetivo: evaluar los factores asociados al autocuidado practicado por personas con estomas de eliminación. Método: estudio transversal analítico, basado en las respuestas de 153 personas con estoma de eliminación, atendidos en un ambulatorio de referencia para la atención de personas con estoma, en la ciudad de Teresina. Se realizó la recolección de datos a través de un formulario semiestructurado con datos demográficos y sobre autocuidado. Los datos se analizaron utilizando estadística descriptiva e inferencial, utilizando la prueba Chi-cuadrado de Pearson. Resultados: vaciar la bolsa, limpiar la estoma, secar la piel periestomal, despegar la placa, medir la estoma, hacer el molde, adaptarlo, vaciar la bolsa, la autoestima y el aislamiento social se asociaron significativamente con el autocuidado (p<0,05). Conclusión: se evidenció que los factores asociados al autocuidado fueron el cuidado de la estoma y del equipo de recolección, además de los impactos en la autoestima y la vida social.

Heavy metals and health risk assessment of Brazilian artisanal cheeses.

Dairy products stand out as a food matrix susceptible to the contamination of heavy metals via cattle feed and environmental or processing conditions. Specifically, in the case of cheese, the concentrations can be further increased depending on the production process. The artisanal cheese market has been standing out, especially in Brazil, due to cultural and gastronomic reasons. Eight types of Brazilian artisanal cheese were analyzed for metal concentrations (chromium, copper, cadmium, lead, arsenic, and mercury, n = 80, 10 samples of each cheese) using inductively coupled plasma mass spectrometry. Based on the results, a health risk assessment was carried out, based on the determination of estimated daily intake, target hazard quotient (THQ), and hazard index (HI). Variable concentrations were observed between the types of cheese, but in all cases the THQ and HI values were less than 1, indicating an absence of potential risk in the consumption of artisanal cheeses in relation to the intake of heavy metals.

Quantitative microbiological risk assessment for the occurrence of listeriosis in Brazil due to the consumption of milk processed by pasteurization or thermosonication.

This study aimed to estimate the risk of listeriosis from the consumption of pasteurized milk in Brazil, comparing conventional treatment with the technology of thermosonication. The Quantitative Microbiological Risk Assessment (QMRA) model was developed, covering the entire milk production chain, from milking to the moment of consumption. In general, higher risks were observed in association with higher initial concentrations of the pathogen and the vulnerable population. The highest risk predicted (3.67 × 10-5) was related to the scenario considering the initial concentration range of L. monocytogenes between 4 and 6 log CFU/mL, with conventional treatment and considering the vulnerable population, resulting in one case of listeriosis every 27,248 servings. When considering thermosonication treatment, lower risks have been predicted. The scenario analysis indicated that the steps related to storage conditions in retail and at the consumer's home (post-processing steps) are the most influential in the associated risk, in all scenarios. The predictive parameters of inactivation related to the applied treatment also have a considerable influence on the risk. The results point to the influence of the stages of the dairy production chain and the thermosonication treatment applied in the food safety of milk, subsidizing information for industrial application and for regulatory agencies.

Watermelon (Citrullus lanatus) rind flour: Development and characterization of a novel watermelon byproduct.

Watermelon (Citrullus lanatus) is a fruit widely consumed by the Brazilian population; however, its rind is usually discarded, despite its nutritional value. This work aimed to develop a watermelon rind flour (WRF) and a fiber-rich bread. The WRF was submitted to microbiological analysis, proximate composition, antioxidant activity, and a profile of phenolic compounds. Six types of bread were developed: three using WRF (20%, 30%, and 40% of wheat flour replacement) and three control samples (only wheat flour), which were submitted to the same analysis, in addition to colorimetric properties, instrumental texture profile, and sensory evaluation (check-all-that-apply, purchase intent, and acceptance test). A high fiber content was observed in WRF (27.15%). The total concentration of phenolic compounds was 2.38 ± 0.20 mg of gallic acid equivalent per g and benzoic acid was the main compound found (73.50 mg/100 g WRF). The results of the antioxidant capacity through the DPPH method indicated a 30% reduction. The WRF 40% bread had 3.06 g of insoluble fiber, characterizing itself as a source of fiber. In the instrumental texture analysis, it was observed that bread hardness increased significantly as the WRF content increased. The formulation of the bread containing WRF is an alternative for the use of waste, favoring the development of a fiber-source product, with a functional food claim.

Prevalence of anti-beta-1 antibody 6 months after hospitalization for acute heart failure predicts adverse outcome.

AIMS: Agonistic antibodies against neurohumoral receptors can induce cardio-noxious effects by altering the baseline receptor activity. To estimate the prevalence of autoantibodies directed against the beta-1 receptor (b1-AAB) in patients admitted to the hospital for acute heart failure (HF) at (i) baseline and (ii) after 6 months of follow-up (F6) and (iii) after another 12 months of follow-up (i.e. 18 months after index hospitalization), to estimate their prognostic impact on clinical outcome (death or first hospitalization for HF). METHODS AND RESULTS: In 47 patients, b1-AAB were serially determined in serum samples collected at index hospitalization and at 6 months of follow-up (F6) with a flow cytometry-based assay: median age 71 years (quartiles 60, 80), 23 (49%) women, 24 (51%) HF with preserved ejection fraction. Beta1-AAB were detected in three subjects at index hospitalization (6%), and in eight subjects at F6 (17%). There were no differences apparent between patients with and without b1-AAB at F6 with regard to age, sex, type, duration, or main cause of HF. During the 12 month period following F6 (i.e. up to month 18), eight events occurred. Event-free survival was associated with prevalence of b1-AAB at F6. Compared with patients without b1-AAB at F6, age-adjusted Cox regression indicated a higher event risk in patients harbouring b1-AAB, with a hazard ratio of 8.96 (95% confidence interval 1.81-44.50, P = 0.007). CONCLUSIONS: Our results suggest a possible adverse prognostic relevance of b1-AAB in patients with acute HF, but this observation needs to be confirmed in larger patient collectives.

Quantitative risk assessment of the presence of fumonisin in corn produced in different regions of Brazil: Influence of climatic conditions.

In this study, the probability of occurrence of fumonisins in corn in the states of greatest production in Brazil was determined. The data were analyzed through quantitative risk analysis using the Monte Carlo simulation. The results indicated that there is a strong correlation between fumonisins contamination levels and the geographical region due to the influence of climatic characteristics, with temperature having the main influence. The Southern states presented higher risks of occurrence and concentration levels of fumonisins in corn due to the temperate climate with lower average temperature and higher relative humidity and precipitation indices. Cultivation in the best season indicates a significant reduction in the production of fumonisins when this period was evaluated, with average concentration levels up to 42% lower. The generated data are important for regulatory agencies and the agricultural sector, which needs to be aware that the chance of success in grain production depends on efficient planning of the growing season, mainly concerning the climatic conditions to which it is subject to minimize the risks.

An interferon gamma response signature links myocardial aging and immunosenescence.

AIMS: Aging entails profound immunological transformations that can impact myocardial homeostasis and predispose to heart failure. However, preclinical research in the immune-cardiology field is mostly conducted in young healthy animals, which potentially weakens its translational relevance. Herein, we sought to investigate how the aging T-cell compartment associates with changes in myocardial cell biology in aged mice. METHODS AND RESULTS: We phenotyped the antigen-experienced effector/memory T cells purified from heart-draining lymph nodes of 2-, 6-, 12-, and 18-month-old C57BL/6J mice using single-cell RNA/T cell receptor sequencing. Simultaneously, we profiled all non-cardiomyocyte cell subsets purified from 2- to 18-month-old hearts and integrated our data with publicly available cardiomyocyte single-cell sequencing datasets. Some of these findings were confirmed at the protein level by flow cytometry. With aging, the heart-draining lymph node and myocardial T cells underwent clonal expansion and exhibited an up-regulated pro-inflammatory transcription signature, marked by an increased interferon-γ (IFN-γ) production. In parallel, all major myocardial cell populations showed increased IFN-γ responsive signature with aging. In the aged cardiomyocytes, a stronger IFN-γ response signature was paralleled by the dampening of expression levels of transcripts related to most metabolic pathways, especially oxidative phosphorylation. Likewise, induced pluripotent stem cells-derived cardiomyocytes exposed to chronic, low grade IFN-γ treatment showed a similar inhibition of metabolic activity. CONCLUSIONS: By investigating the paired age-related alterations in the T cells found in the heart and its draining lymph nodes, we provide evidence for increased myocardial IFN-γ signaling with age, which is associated with inflammatory and metabolic shifts typically seen in heart failure.

Cold plasma as a pre-treatment for processing improvement in food: A review.

Thermal processes can be very damaging to the nutritional and sensory quality of foods. Non-thermal technologies have been applied to reduce the impact of heat on food, reducing processing time and increasing its efficiency. Among many non-thermal technologies, cold plasma is an emerging technology with several potential applications in food processing. This technique can be used to preserve and sanitize food products, and act as a pre-treatment for drying, extraction, cooking, curing, and hydrogenation of foods. Furthermore, the reacting plasma species formed during the plasma application can change positively the sensory and nutritional aspects of foods. The aim of this review is to analyze the main findings on the application of cold plasma as a pre-treatment technology to improve food processing. In its current maturity stage, the cold plasma technology is suitable for reducing drying time, increasing extraction efficiency, as well as curing meats. This technology can convert unsaturated into saturated fats, without forming trans isomers, which can be an alternative to healthier foods. Although many advantages come from cold plasma applications, this technology still has several challenges, such as the scaling up, especially in increasing productivity and treating foods with large formats. Optimization and control of the effects of plasma on nutritional and sensory quality are still under investigation. Further improvement of the technology will come with a higher knowledge of the effects of plasma on the different chemical groups present in foods, and with the development of bigger or more powerful plasma systems.

Platelet-Derived MicroRNAs Regulate Cardiac Remodeling After Myocardial Ischemia.

BACKGROUND: Platelets can infiltrate ischemic myocardium and are increasingly recognized as critical regulators of inflammatory processes during myocardial ischemia and reperfusion (I/R). Platelets contain a broad repertoire of microRNAs (miRNAs), which, under certain conditions such as myocardial ischemia, may be transferred to surrounding cells or released into the microenvironment. Recent studies could demonstrate that platelets contribute substantially to the circulating miRNA pool holding the potential for so far undiscovered regulatory functions. The present study aimed to determine the role of platelet-derived miRNAs in myocardial injury and repair following myocardial I/R. METHODS: In vivo model of myocardial I/R, multimodal in vivo and ex vivo imaging approaches (light-sheet fluorescence microscopy, positron emission tomography and magnetic resonance imaging, speckle-tracking echocardiography) of myocardial inflammation and remodeling, and next-generation deep sequencing analysis of platelet miRNA expression. RESULTS: In mice with a megakaryocyte/platelet-specific knockout of pre-miRNA processing ribonuclease Dicer, the present study discloses a key role of platelet-derived miRNAs in the tightly regulated cellular processes orchestrating left ventricular remodeling after myocardial I/R following transient left coronary artery ligation. Disruption of the miRNA processing machinery in platelets by deletion of Dicer resulted in increased myocardial inflammation, impaired angiogenesis, and accelerated development of cardiac fibrosis, culminating in an increased infarct size by d7 that persisted through d28 of myocardial I/R. Worsened cardiac remodeling after myocardial infarction in mice with a platelet-specific Dicer deletion resulted in an increased fibrotic scar formation and distinguishably increased perfusion defect of the apical and anterolateral wall at day 28 post-myocardial infarction. Altogether, these observations culminated in an impaired left ventricular function and hampered long-term cardiac recovery after experimental myocardial infarction and reperfusion therapy. Treatment with the P2Y12 (P2Y purinoceptor 12) antagonist ticagrelor completely reversed increased myocardial damage and adverse cardiac remodeling observed in DicerPf4∆/Pf4∆ mice. CONCLUSIONS: The present study discloses a critical role of platelet-derived miRNA in myocardial inflammation and structural remodeling processes following myocardial I/R.

El cine durante la cuarentena: Una herramienta pedagógica de bioética en estudiantes de posgrado en salud / The Cinema During the Quarantine: a Pedagogical Tool of Bioethics in Postgraduate Health Students

La bioética estudia la conducta humana en los campos de las ciencias biológicas y de la atención de la salud. Es imprescindible que el contenido programático de las facultades de ciencias de la salud divulgue su significado e importancia. Todo aquel que pretenda una práctica profesional en la cual interaccione directamente con la vida, debe adquirir competencias desde el campo de la bioética con fundamentos arraigados en los principios éticos de no maleficencia, beneficencia, autonomía y justicia. De tal forma que desarrollen capacidad de reflexión crítica sobre los conflictos éticos provocados por los avances de la ciencia de la vida y la medicina. Cada día se propone más el uso de herramientas audiovisuales en la docencia, para facilitar el abordaje de dilemas éticos, crear espacios de análisis, discusión, que ayudan a la comprensión, memoria e interpretación de este tema entre los estudiantes. En este artículo se pretende divulgar la experiencia a través de una estrategia pedagógica que involucra el cineforo como medio de aprendizaje en bioética en un grupo de estudiantes posgrado en salud. (AU)

Bioethics studies human behavior in the field of biological sciences and health care. It is essential that the programmatic content of the faculties of health sciences disclose its meaning and importance. Anyone who intends a professional practice, in which they interact directly with life, must acquire skills from the field of bioethics with foundations rooted in the ethical principles of non-maleficence, beneficence, autonomy and justice. In such a way that they develop the capacity for critical reflection on the ethical conflicts caused by advances in the science of life and medicine. Every day, the use of audiovisual tools in teaching is proposed more, to make it easier for students to approach ethical dilemmas and create spaces for analysis, discussion, help understanding, memory and interpretation of this topic. This article aims to disseminate the experience through the implementation of a pedagogical strategy that involves cinema and commercial films as a means of learning bioethics in a group of postgraduate health students. (AU)

Characterization of the effect of the GLUT-1 inhibitor BAY-876 on T cells and macrophages.

Increased aerobic glycolysis is a metabolic hallmark of proinflammatory leukocytes including macrophages and T cells. To take up glucose from the environment and fuel glycolysis, activated leukocytes upregulate the glucose transporter GLUT1. The orally bioavailable selective GLUT1 inhibitor BAY-876 was developed primarily as an anti-tumor drug. Our study assessed its activity on activated macrophages and CD4+ T cells. BAY-876 significantly attenuated glucose uptake by cultured CD4+ T cells and macrophages by 41% and 15%, respectively. Extracellular flux analysis of activated CD4+ T cells in vitro showed that BAY-876 significantly decreases glycolytic proton flux rate and lactate production, effects that are accompanied by an increased oxidative phosphorylation-mediated ATP production rate, leaving intracellular ATP levels per cell unchanged. However, GLUT1 inhibition reduced CD4+ T cell proliferation without compromising cell viability and reduced IFN-γ secretion by 20%. Moreover, TNF secretion from macrophages was reduced by 27%. We conclude that GLUT1-specific inhibitors, like BAY-876, deserve further in vivo testing in a broad range of (auto-) inflammatory disease models.

Myocardial Milieu Favors Local Differentiation of Regulatory T Cells.

BACKGROUND: In the past years, several studies investigated how distinct immune cell subsets affects post-myocardial infarction repair. However, whether and how the tissue environment controls these local immune responses has remained poorly understood. We sought to investigate how antigen-specific T-helper cells differentiate under myocardial milieu's influence. METHODS: We used a transgenic T cell receptor (TCR-M) model and major histocompatibility complex-II tetramers, both myosin-specific, combined with single-cell transcriptomics (single-cell RNA sequencing [scRNA-seq]) and functional phenotyping to elucidate how the antigen-specific CD4+ T cells differentiate in the murine infarcted myocardium and influence tissue repair. Additionally, we transferred proinflammatory versus regulatory predifferentiated TCR-M-cells to dissect how they specially contribute to post-myocardial infarction inflammation. RESULTS: Flow cytometry and scRNA-/TCR-seq analyses revealed that transferred TCR-M cells acquired an induced regulatory phenotype (induced regulatory T cell) in the infarcted myocardium and blunted local inflammation. Myocardial TCR-M cells differentiated into 2 main lineages enriched with either cell activation and profibrotic transcripts (eg, Tgfb1) or suppressor immune checkpoints (eg, Pdcd1), which we also found in human myocardial tissue. These cells produced high levels of LAP (latency-associated peptide) and inhibited IL-17 (interleukin-17) responses. Endogenous myosin-specific T-helper cells, identified using genetically barcoded tetramers, also accumulated in infarcted hearts and exhibited a regulatory phenotype. Notably, TCR-M cells that were predifferentiated toward a regulatory phenotype in vitro maintained stable in vivo FOXP3 (Forkhead box P3) expression and anti-inflammatory activity whereas TH17 partially converted toward a regulatory phenotype in the injured myocardium. Overall, the myosin-specific Tregs dampened post-myocardial infarction inflammation, suppressed neighboring T cells, and were associated with improved cardiac function. CONCLUSIONS: These findings provide novel evidence that the heart and its draining lymph nodes actively shape local immune responses by promoting the differentiation of antigen-specific Tregs poised with suppressive function.

Thermosonication as a pretreatment of raw milk for Minas frescal cheese production.

Minas frescal cheese is extremely popular in Brazil, with high perishability and acceptability. Among emerging technologies, ultrasound stands out for its satisfactory results regarding microbiological safety and technological and sensory aspects. The combined mild temperature application, called thermosonication, can generate even more promising results. In this study, a high-intensity ultrasound system combined with thermal heating (TS, thermosonication) was applied for the treatment of raw milk to produce Minas Frescal cheese. US energy was delivered to raw milk samples using a probe operating at a 20 kHz of frequency and nominal power of 160, 400, and 640 W. The TS system was compared with conventional pasteurization (HTST, high-temperature short-time pasteurization) at 72 to 75 °C and 15 s. Soft cheeses were prepared with different samples: (a) raw milk (control), b)conventionally pasteurized milk (HTST), and c) TS treat milk in different nominal power (TS160, TS400, and TS640). The produced cheeses were evaluated for microbiological behavior, rheology, color parameters, and bioactive compounds. TS treatment in milk resulted in higher microbial inactivation and stability during storage, improved color parameters (higher lightness (L*), and whiteness index (WI). TS treatment also showed a higher generation of bioactive compounds (higher antioxidant, and inhibitory activities of α-amylase, α-glucosidase, and angiotensin-converting enzymes) than HTST. The impact of TS on rheological properties was similar to HTST, resulting in more brittle and less firm products than the cheese produced with raw milk. The positive effects were more prominent using a nominal power of 400 W (TS400). Therefore, TS proved to be a promising process for processing milk for Minas Frescal cheese production.

Extracellular Matrix Protein-1 as a Mediator of Inflammation-Induced Fibrosis After Myocardial Infarction.

Irreversible fibrosis is a hallmark of myocardial infarction (MI) and heart failure. Extracellular matrix protein-1 (ECM-1) is up-regulated in these hearts, localized to fibrotic, inflammatory, and perivascular areas. ECM-1 originates predominantly from fibroblasts, macrophages, and pericytes/vascular cells in uninjured human and mouse hearts, and from M1 and M2 macrophages and myofibroblasts after MI. ECM-1 stimulates fibroblast-to-myofibroblast transition, up-regulates key fibrotic and inflammatory pathways, and inhibits cardiac fibroblast migration. ECM-1 binds HuCFb cell surface receptor LRP1, and LRP1 inhibition blocks ECM-1 from stimulating fibroblast-to-myofibroblast transition, confirming a novel ECM-1-LRP1 fibrotic signaling axis. ECM-1 may represent a novel mechanism facilitating inflammation-fibrosis crosstalk.