Blockage of the adenosine A2B receptor prevents cardiac fibroblasts overgrowth in rats with pulmonary arterial hypertension.

Sustained pressure overload and fibrosis of the right ventricle (RV) are the leading causes of mortality in pulmonary arterial hypertension (PAH). Although the role of adenosine in PAH has been attributed to the control of pulmonary vascular tone, cardiac reserve, and inflammatory processes, the involvement of the nucleoside in RV remodelling remains poorly understood. Conflicting results exist on targeting the low-affinity adenosine A2B receptor (A2BAR) for the treatment of PAH mostly because it displays dual roles in acute vs. chronic lung diseases. Herein, we investigated the role of the A2BAR in the viability/proliferation and collagen production by cardiac fibroblasts (CFs) isolated from RVs of rats with monocrotaline (MCT)-induced PAH. CFs from MCT-treated rats display higher cell viability/proliferation capacity and overexpress A2BAR compared to the cells from healthy littermates. The enzymatically stable adenosine analogue, 5'-N-ethylcarboxamidoadenosine (NECA, 1-30 µM), concentration-dependently increased growth, and type I collagen production by CFs originated from control and PAH rats, but its effects were more prominent in cells from rats with PAH. Blockage of the A2BAR with PSB603 (100 nM), but not of the A2AAR with SCH442416 (100 nM), attenuated the proliferative effect of NECA in CFs from PAH rats. The A2AAR agonist, CGS21680 (3 and 10 nM), was virtually devoid of effect. Overall, data suggest that adenosine signalling via A2BAR may contribute to RV overgrowth secondary to PAH. Therefore, blockage of the A2AAR may be a valuable therapeutic alternative to mitigate cardiac remodelling and prevent right heart failure in PAH patients.

Designing a youth-led Dialogue Forum tool: The CO-CREATE experience.

While obesity prevention represents an established field of research, the inclusion of young people, who are regularly cited as an important priority group, are rarely actioned in long-term studies. This paper focuses on the development of a dialogue tool intended to tackle this issue, engaging, and eliciting insights on the theme of obesity prevention, by young people and for young people. As part of the CO-CREATE project, this tool was co-developed by designers, public health, and youth participation experts, researchers, and young people. Co-creation is a key methodology in the design of the dialogue tool, as young people were involved in all stages of the development process. This paper elaborates on the process of co-designing a dialogue tool that helps explore obesity prevention policy ideas from multiple perspectives, and describes the design principles that informed the process and the final versions of the tool. The purpose of the Dialogue Forum tool is for youth to engage policymakers and other relevant stakeholders to discuss and refine co-created and youth-initiated ideas for healthier food and physical activity environments. We analyze how specific design principles were integrated into different prototypes and the value of this within the project and the field.

A bioinspired multifunctional hydrogel patch targeting inflammation and regeneration in chronic intestinal wounds.

Healing of intestinal chronic wounds remains a major challenge as current therapies are ineffective in promoting proper regeneration of the damaged intestinal wall. An innovative concept, based on a bioinspired multifunctional alginate-melanin hybrid 3D scaffold, to target both inflammatory and regenerative processes, is proposed herein. Hydrogel-entrapped melanin nanoparticles demonstrated free-radical scavenging activity, supported by the neutralization of free-radicals in solution (90%), and the in vitro capture of reactive oxygen species (ROS) produced by stimulated macrophages in an inflammatory-mimicking environment. Notably, scaffolds could be reused (at least 3 times), while maintaining these properties. The extracellular matrix (ECM)-inspired biomaterial, containing protease-sensitive and integrin-binding domains, exhibited remarkable ability for cell colonisation. Human intestinal fibroblasts and epithelial cells (Caco-2) co-seeded on lyophilized scaffolds were able to invade/colonize the construct and produce endogenous ECM, key for neo-tissue formation and re-epithelialization. Scaffolds presented tuneable mechanical properties and could be used both in hydrated and freeze-dried states, maintaining their performance upon rehydration, which are attractive features for clinical application. Collectively, our results highlight the potential of biofunctionalized alginate-melanin hybrid 3D scaffolds as multi-therapeutic patches for modulating inflammation and tissue regeneration in chronic intestinal wounds, which address a major but still unmet clinical need. The proposed multi-therapeutic strategy may potentially be extended to the treatment of other chronic wounds.

Fibrinogen and magnesium combination biomaterials modulate macrophage phenotype, NF-kB signaling and crosstalk with mesenchymal stem/stromal cells.

Macrophage behavior upon biomaterial implantation conditions the inflammatory response and subsequent tissue repair. The hypothesis behind this work was that fibrinogen (Fg) and magnesium (Mg) biomaterials, used in combination (FgMg) could act synergistically to modulate macrophage activation, promoting a pro-regenerative phenotype. Materials were characterized by scanning electron microscopy, Fg and Mg degradation products were quantified by atomic absorption spectroscopy and ELISA. Whole blood immune cells and primary human monocyte-derived macrophages were exposed to the biomaterials extracts in unstimulated (M0) or pro-inflammatory LPS or LPS-IFNγ (M1) conditions. Macrophage phenotype was evaluated by flow cytometry, cytokines secreted by whole blood cells and macrophages were measured by ELISA, and signaling pathways were probed by Western blotting. The secretomes of macrophages preconditioned with biomaterials extracts were incubated with human mesenchymal stem/stromal cells (MSC) and their effect on osteogenic differentiation was evaluated via Alkaline Phosphatase (ALP) activity and alizarin red staining. Scaffolds of Fg, alone or in the FgMg combination, presented similar 3D porous architectures. Extracts from FgMg materials reduced LPS-induced TNF-α secretion by innate immune cells, and macrophage M1 polarization upon LPS-IFNγ stimulation, resulting in lower cell surface CD86 expression, lower NFκB p65 phosphorylation and reduced TNF-α secretion. Moreover, while biomaterial extracts per se did not enhance MSC osteogenic differentiation, macrophage secretome, particularly from cells exposed to FgMg extracts, increased MSC ALP activity and alizarin red staining, compared with extracts alone. These findings suggest that the combination of Fg and Mg synergistically influences macrophage pro-inflammatory activation and crosstalk with MSC. STATEMENT OF SIGNIFICANCE: Modulating macrophage phenotype by degradable and bioactive biomaterials is an increasingly explored strategy to promote tissue repair/regeneration. Fibrinogen (Fg) and magnesium (Mg)-based materials have been explored in this context. Previous work from our group showed that monocytes interact with fibrinogen adsorbed onto chitosan surfaces through TLR4 and that fibrinogen scaffolds promote in vivo bone regeneration. Also, magnesium ions have been reported to modulate macrophage pro-inflammatory M1 stimulation and to promote bone repair. Here we report, for the first time, the combination of Fg and Mg materials, hypothesizing that it could act synergistically on macrophages, directing them towards a pro-regenerative phenotype. As a first step towards proving/disproving our hypothesis we used extracts obtained from Fg, Mg and FgMg multilayer constructs. We observed that FgMg extracts led to a reduction in the polarization of macrophages towards a pro-inflammatory phenotype. Also, the secretome of macrophages exposed to extracts of the combination material promoted the expression of osteogenic markers by MSCs.

Osteoclasts degrade fibrinogen scaffolds and induce mesenchymal stem/stromal osteogenic differentiation.

Fibrinogen (Fg) is a pro-inflammatory protein with pro-healing properties. Previous work showed that fibrinogen 3D scaffolds (Fg-3D) promote bone regeneration, but the cellular players were not identified. Osteoclasts are bone resorbing cells that promote bone remodeling in close crosstalk with osteoblasts. Herein, the capacity of osteoclasts differentiated on Fg-3D to degrade the scaffolds and promote osteoblast differentiation was evaluated in vitro. Fg-3D scaffolds were prepared by freeze-drying and osteoclasts were differentiated from primary human peripheral blood monocytes. Results obtained showed osteoclasts expressing the enzymes cathepsin K and tartrate resistant acid phosphatase colonizing Fg-3D scaffolds. Osteoclasts were able to significantly degrade Fg-3D, reducing the scaffold's area, and increasing D-dimer concentration, a Fg degradation product, in their culture media. Osteoclast conditioned media from the first week of differentiation promoted significantly stronger human primary mesenchymal stem/stromal cell (MSC) osteogenic differentiation, evaluated by alkaline phosphatase activity. Moreover, week 1 osteoclast conditioned media promoted earlier MSC osteogenic differentiation, than chemical osteogenesis inductors. TGF-ß1 was found increased in osteoclast conditioned media from week 1, when compared to week 3 of differentiation. Taken together, our results suggest that osteoclasts are able to differentiate and degrade Fg-3D, producing factors like TGF-ß1 that promote MSC osteogenic differentiation.

The Systemic Immune Response to Collagen-Induced Arthritis and the Impact of Bone Injury in Inflammatory Conditions.

Rheumatoid arthritis (RA) is a systemic disease that affects the osteoarticular system, associated with bone fragility and increased risk of fractures. Herein, we aimed to characterize the systemic impact of the rat collagen-induced arthritis (CIA) model and explore its combination with femoral bone defect (FD). The impact of CIA on endogenous mesenchymal stem/stromal cells (MSC) was also investigated. CIA induction led to enlarged, more proliferative, spleen and draining lymph nodes, with altered proportion of lymphoid populations. Upon FD, CIA animals increased the systemic myeloid cell proportions, and their expression of co-stimulatory molecules CD40 and CD86. Screening plasma cytokine/chemokine levels showed increased tumor necrosis factor-α (TNF-α), Interleukin (IL)-17, IL-4, IL-5, and IL-12 in CIA, and IL-2 and IL-6 increased in CIA and CIA+FD, while Fractalkine and Leptin were decreased in both groups. CIA-derived MSC showed lower metabolic activity and proliferation, and significantly increased osteogenic and chondrogenic differentiation markers. Exposure of control-MSC to TNF-α partially mimicked the CIA-MSC phenotype in vitro. In conclusion, inflammatory conditions of CIA led to alterations in systemic immune cell proportions, circulating mediators, and in endogenous MSC. CIA animals respond to FD, and the combined model can be used to study the mechanisms of bone repair in inflammatory conditions.

Macrophages Down-Regulate Gene Expression of Intervertebral Disc Degenerative Markers Under a Pro-inflammatory Microenvironment.

Low back pain is a highly prevalent clinical problem and intervertebral disc (IVD) degeneration is now accepted as the major pathophysiological mechanism responsible for this condition. Accumulating evidence suggests that inflammation plays a crucial role in the progression of human IVD degeneration, with macrophages being pointed as the key immune cell players in this process since their infiltration in degenerated IVD samples has been extensively demonstrated. Since they are highly plastic, macrophages can play different roles depending on the microenvironmental cues. The study of inflammation associated with IVD degeneration has been somehow neglected and one of the reasons is related with lack of adequate models. To overcome this, we established and characterized a new model of IVD organ culture under pro-inflammatory conditions to further dissect the role of macrophages in IVD associated immune response. For that, human monocyte-derived macrophages were co-cultured either with bovine caudal IVD punches in the presence of the pro-inflammatory cytokine IL-1ß, or IVD-conditioned medium (CM), to investigate how IVD-produced factors influence macrophage phenotype. After 72 h, metabolic activity, gene expression and cytokine profile of macrophages and IVD cells were measured. Our results show that macrophages and IVDs remain metabolically active in the presence of IL-1ß, significantly upregulate CCR7 gene expression and increase production of IL-6 on macrophages. When treating macrophages with IL-1ß-IVD-CM, CCR7 upregulation follows the same trend, while for IL-6 an opposite effect was observed. On the other hand, macrophages interfere with IVD ECM remodeling, decreasing MMP3 expression and downregulating aggrecan and collagen II gene expression in the presence of IL-1ß. Overall, the co-culture model established in this study can be considered a suitable approach to address the cellular and molecular pathways that regulate macrophage-IVD crosstalk, suggesting that degenerated IVD tissue tends to polarize human macrophages toward a more pro-inflammatory profile, which seems to aggravate IVD degeneration. This model could be used to improve the knowledge of the mechanisms that link IVD degeneration and the immune response.

Impact of physical activity in vascular cognitive impairment (AFIVASC): study protocol for a randomised controlled trial.

BACKGROUND: Cognitive impairment and cerebrovascular pathology are both frequent with ageing. Cognitive impairment due to vascular pathology of the brain, termed vascular cognitive impairment (VCI), is one of the most frequent causes of cognitive impairment in elderly subjects. Thus far, VCI has no specific pharmacological treatment. Recent observational studies have suggested a protective effect of physical activity in cognition, but adequate randomised controlled trials (RCT) are lacking. METHODS: AFIVASC is a multi-centre randomised controlled trial, with a 6-month intervention treatment and an additional follow-up of 6 months, that aims to estimate the impact of 6 months of moderate intensity physical activity on cognition (the primary outcome) at 6 and 12 months in subjects with VCI. Participants are community dwellers with criteria for VCI without dementia or who have had previous stroke or transient ischaemic attack (TIA). Patients may be self-referred or referred from a medical appointment. After confirming the inclusion criteria, a run-in period of 1 month is conducted to access adherence; only after that are subjects randomly assigned (using a computerised program blinded to clinical details) to two groups (intervention group and best practice usual care group). The intervention consists of three physical activity sessions of 60 min each (two supervised and one unsupervised) per week. The primary outcome is measured by the presence or absence of decline in cognitive status. Secondary outcomes include changes in neuro-cognitive measures, quality of life, and functional and motor status. Primary and secondary outcomes are evaluated at 6 and 12 months by investigators blinded to both intervention and randomisation. A required sample size of 280 subjects was estimated. Statistical analyses will include regression analysis with repeated measures. The study was approved by the Ethics Committee for Health of Centro Hospitalar de Lisboa Norte (ref. no. 1063/13) and by the Ethics Committee for Health of Centro Hospitalar do Porto CHP (ref. no. 2016.055(049-DEFI/048-CES)). DISCUSSION: We aim to show whether or not moderate physical activity has a beneficial impact on cognition, quality of life, motor, and functional status in people with vascular cognitive impairment, and to generate new insights on the applicability of implementing physical activity in this specific population. TRIAL REGISTRATION: ClinicalTrials.gov, NCT03578614 July 6, 2018.

Programa de intervenção comunitária: "A outra face do cuidar" / Community intervention program: "The other face of caring" / Programa de intervención comunitaria: "La otra cara de la atención"

Introdução: O cuidador informal (CI) presta cuidados a pessoas dependentes, não sendo recompensado economicamente. São inúmeros os fatores de estresse inerentes à tarefa de cuidar, com consequente desgaste físico e emocional. Estudos indicam que as abordagens que englobam a realização de atividade física apresentam um impacto positivo na diminuição da depressão, estresse, raiva e sobrecarga apresentados pelo CI. Objetivo: Reduzir a sobrecarga do CI, com vista à capacitação da tarefa de cuidar, pela aquisição de ferramentas na gestão da sobrecarga. Métodos: De uma listagem de 90 utentes com a codificação "demência" e 50 com "atraso mental" realizou-se o diagnóstico de situação através da aplicação da escala de Zarit que avalia a sobrecarga dos CI. Destes, 13 mostraram interesse em participar na intervenção, que consistiu em sessões de educação para a saúde, sessões de yoga e dinamização de um grupo de apoio. Foi avaliado o grau de sobrecarga dos participantes e a sua satisfação. Resultados: Os CI eram maioritariamente mulheres (92%), com idade média de 62 anos. Foram realizadas 100% das sessões previstas, com 23% de desistência dos participantes. A sobrecarga moderada a severa ou severa diminuiu de 92.3% para 60% no final da intervenção. Todos os CI classificaram o projeto como "bom" ou "muito bom". Discussão: O projeto cumpriu a planificação inicial, tendo-se verificado uma diminuição do grau de sobrecarga do CI como evidenciado em outros estudos. Destaca-se como limitação a dificuldade do CI na realização de outras atividades além de cuidar do seu dependente. Conclusão: A intervenção teve um impacto positivo nos CI, tendo cumprido os objetivos inicialmente propostos. A equipa de saúde deu continuidade a este projeto através de uma parceria com uma cooperativa de solidariedade social local.

Introduction: The informal caregiver (IC) provides assistance to disabled people without any financial gratification. Many stress factors are associated with the task of taking care of others, leading to physical and emotional deterioration. Studies indicate that approaches involving physical activity have a positive impact on the reduction of depression, stress, anger and overload presented by the IC. Objective: To reduce IC overload with a view to enabling the task of caring by acquiring tools for overload management. Methods: The data collection of ICs overload was made using Zarit scale from a previous database of 90 patients classified as "dementia" and 50 as "mental retardation". 13 IC's express their interest to be part of the project. The intervention consisted in health education sessions; yoga classes; establishment of a support group. The participant's degree of overload and their satisfaction were assessed. Results: IC were mainly women (92%), with average age of 62 years. All scheduled sessions took place, with 23% participants withdrawing. Moderate to severe or severe overload level reduced from 92.3% to 60% at the end of the intervention and all participants classified the project as "good" or "very good". Discussion: The project fulfilled the initial planning. Decrease of the IC overload was noted as evidenced in other studies. The limitation is the difficulty of IC in performing other activities besides then taking care of its dependent. Conclusions: The intervention had a positive impact in the ICs and the main goals were achieved. The health team continued this project through a partnership with a local social solidarity cooperative.

Introducción: El cuidador informal (CI) presta cuidados a las personas dependientes, no siendo recompensado económicamente. Son innumerables los factores de estrés inherentes a la tarea de cuidar, con consiguiente desgaste físico y emocional. Los estudios indican que los enfoques que incluyen la actividad física tienen un impacto positivo en la reducción de la depresión, el estrés, la ira y la sobrecarga que presenta el CI. Objetivo: Reducir la sobrecarga del CI, con miras a la capacitación de la tarea de cuidar y la adquisición de herramientas en la gestión de la sobrecarga. Métodos: De una lista de 90 pacientes con la clasificación "demencia" y 50 con "retraso mental" se realizó el diagnóstico de sobrecarga del CI, aplicándose la escala de Zarit. De ellos, 13 mostraron interés en participar en la intervención, que consistió en sesiones de educación para la salud, sesiones de yoga y dinamización de un Grupo de Apoyo. Se evaluó el grado de sobrecarga de los participantes y su satisfacción. Resultados: Los CI fueron en su mayoría mujeres (92%), con una edad promedio de 62 años. El 100% de las sesiones se llevaron a cabo, y el 23% de los participantes se rindieron. La sobrecarga moderada a severa o severa disminuyó del 92,3% al 60% al final de la intervención. Todos los CIs calificaron el proyecto como "bueno" o "muy bueno". Discusión: El proyecto cumplió la planificación inicial, habiéndose comprobado una disminución del grado de sobrecarga del CI como lo demuestran otros estudios. Se destaca como limitación la dificultad del CI en la realización de otras actividades además de cuidar de su dependiente. Conclusión: La intervención tuvo un impacto positivo en los CI, habiendo cumplido los objetivos inicialmente propuestos. El equipo de salud continuó este proyecto a través de una asociación con una cooperativa de solidaridad social local.

A 5-Year Follow-Up of The Benefits of an Exercise Training Program in Liver Recipients Transplanted Due to Familial Amyloidotic Polyneuropathy.

BACKGROUND: Supervised (SE) and home-based exercise (HBE) training regimes are effective on reconditioning patients with familial amyloidotic polyneuropathy (FAP) after liver transplantation, but research of the long-term retention of the benefits attained in patients with FAP has not yet been conducted. PURPOSE: In this 5-year follow-up study, we aimed to determine whether the exercise training gains in body composition, physical activity, and function promoted by a 24-week SE or HBE training regimes are retained in patients with FAP who resume normal activity. METHODOLOGY: Sixteen liver-transplanted patients with FAP were reassessed for body composition (dual X-ray absorptiometry), physical activity (questionnaire), and function (handgrip strength and 6-minute walk test). RESULTS: Total body fat increased with both exercise regimes during follow-up ( P < .05; η2 = 0.432-0.625) as well as femoral neck bone density ( P = .048; η2 = 0.119). However, gains in upper limbs muscle quality during follow-up ( P < .001; η2 = 0.597) were only found in the SE group ( P = .042; η2 = 0.245). Both exercise regimes showed retaining aptitudes in walking capacity ( P < .05; η2 = 0.329-0.460) and muscle mass ( P = .05; η2 = 0.245). Still, none could retain the physical activity levels. CONCLUSION: Long-term resumption of normal activity following a 24-week SE or HBE regime in patients with FAP resulted in loss of exercise induced increases in physical activity but counterweighted postoperative losses in bone mineral density and substantially retained the benefits in walking capacity, muscle mass, and quality, in particular, in the SE group.

Is the adenosine A2B 'biased' receptor a valuable target for the treatment of pulmonary arterial hypertension?

Pulmonary arterial hypertension (PAH) is a maladaptive disorder characterized by increased pulmonary vascular resistance leading to right ventricular failure and death. Adenosine released by injured tissues, such as the lung and heart, influences tissue remodeling through the activation of adenosine receptors. Evidence regarding activation of the low-affinity A2BAR by adenosine points towards pivotal roles of this receptor in processes associated with both acute and chronic lung diseases. Conflicting results exist concerning the beneficial or detrimental roles of the A2B 'biased' receptor in right ventricular failure secondary to PAH. In this review, we discuss the pros and cons of manipulating A2BARs as a putative therapeutic target in PAH.

Três médicos e uma mãe / Three doctors and a mother / Tres médicos y una madre

A papilomatose laríngea, doença rara potencialmente fatal, carateriza-se pela proliferação de papilomas no trato respiratório, múltiplos, recorrentes, cuja etiologia é a infeção por papilomavírus humano (HPV). Menina, 21 meses, filha de mãe com serologia positiva para vírus da imunodeficiência humana (VIH) e HPV. Em acompanhamento nas consultas de Pediatria do Desenvolvimento do Hospital, Pediatra Particular e Médico de Família (MF). Aos 18 meses, na consulta de acompanhamento do MF, a mãe preocupada salienta a fala sussurrada e choro rouco da filha com diagnóstico frequente de laringite aguda no Pediatra e MF nos últimos 3 meses, motivando a referenciação à Otorrinolaringologia e posterior diagnóstico de papilomatose laríngea. A abordagem da disfonia na criança evita o uso inapropriado de corticoides, inibidores da bomba de prótons e antibioticoterapia. Neste relato sobressai a desvantagem associada ao seguimento por múltiplos médicos, sendo o MF fundamental para reunir e integrar a informação clínica, permitindo a continuidade de cuidados.

Laryngeal papillomatosis is a rare and potentially fatal disease characterized by the proliferation of recurrent respiratory papillomas, whose etiology is human papillomavirus (HPV) infection. We report a clinical case of a 21-month girl, whose mother is sero-positive to human immunodeficiency virus (HIV) infection and HPV. This girl attended multiple medical consultations: Development Pediatrics (at the hospital), private Pediatrician and General Practitioner (GP). At 18 months, in the GP surveillance consultation, the concerned mother referred whispered talking, hoarse crying and frequent diagnosis of acute laryngitis at the Pediatrician in the last 3 months. She was referenced to otorhinolaryngology with subsequent diagnosis of laryngeal papillomatosis. The approach to childhood dysphonia avoids inappropriate use of corticosteroids, proton pump inhibitors and antibiotics. This report highlights the disadvantage of the surveillance by multiple doctors and the key role of the GP in gathering and integrating clinical information, allowing the continuity of care.

La papilomatosis laríngea, una enfermedad rara y potencialmente mortal, se caracteriza por la proliferación de papilomas respiratorios recurrentes, cuya etiología es la infección por el virus del papiloma humano (VPH). Se relata el caso de una niña de 21 meses, hija de una madre seropositiva al virus de la inmunodeficiencia humana (VIH) y VPH. Vigilada en las consultas de Pediatría de Desarrollo del Hospital, Pediatría Privada y Médico de la Familia (MF). A los 18 meses, en la consulta de vigilancia del MF, la madre preocupada destaca habla susurrada, llanto ronco y diagnóstico frecuente de la laringitis aguda en la pediatra en los últimos 3 meses. Se referenció a otorrinolaringología con posterior diagnóstico de papilomatosis laríngea. El enfoque de la disfonía infantil evita el uso inapropiado de los corticosteroides, inhibidores de la bomba de protones y antibioterapia. En este informe se destaca la desventaja asociada al seguimiento por parte de varios médicos, y el papel clave del MF para reunir e integrar la información clínica, lo que permite la continuidad de la atención.