An affinity-based approach to engineer laminin-presenting cell instructive microenvironments

Laminin immobilization into diverse biological and synthetic matrices has been explored to replicate the microenvironment of stem cell niches and gain insight into the role of extracellular matrix (ECM) on stem cell behavior. However, the site-specific immobilization of this heterotrimeric glycoprotein and, consequently, control over its orientation and bioactivity has been a challenge that has limited many of the explored strategies to date. In this work, we established an affinity-based approach that takes advantage of the native high affinity interaction between laminin and the human N-terminal agrin (hNtA) domain. This interaction is expected to promote the site-selective immobilization of laminin to a specific substrate, while preserving the exposure of its key bioactive epitopes. Recombinant hNtA (rhNtA) domain was produced with high purity (>90%) and successfully conjugated at its N-terminal with a thiol-terminated poly(ethylene glycol) (PEG) without affecting its affinity to laminin. Self-assembled monolayers (SAMs) of mono-PEGylated rhNtA on gold (mPEG rhNtA-SAMs) were then prepared to evaluate the effectiveness of this strategy. The site-specific immobilization of laminin onto mPEG rhNtA-SAMs was shown to better preserve protein bioactivity in comparison to laminin immobilized on SAMs of thiol-PEG-succinimidyl glutaramide (HS-PEG-SGA), used for the non-selective covalent immobilization of laminin, as evidenced by its enhanced ability to efficiently self-polymerize and mediate cell adhesion and spreading of human neural stem cells. These results highlight the potential of this novel strategy to be used as an alternative to the conventional immobilization approaches in a wide range of applications, including engineered coatings for neuroelectrodes and cell culture, as well as biofunctionalization of 3D matrices.The mass spectrometry technique was performed at the Proteomics i3S Scientific Platform ( Roteiro/0028/2013 ; LISBOA-01-0145-FEDER-022125 ) with the assistance of Hugo Osório. IRRAS analysis was performed at the Biointerfaces and Nanotechnology Scientific Platform, with the assistance of Ricardo Vidal. In Cell Analyzer experiments were performed at the Biosciences Screening i3S Scientific Platform , member of the PPBI ( PPBI-POCI-01-0145-FEDER-022122 ) with the assistance of André Maia. Confocal microscopy was conducted at the Bioimaging i3S Scientific Platform , member of the PPBI ( PPBI-POCI-01-0145-FEDER-022122 ), with the assistance of Maria Lázaro. This work was supported by projects NORTE-01-0145-FEDER-000008 and NORTE-01-0145-FEDER-000012 , supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF) and FEDER - Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation ( POCI ), Portugal 2020; by Portuguese funds through FCT/MCTES in the framework of the project “Institute for Research and Innovation in Health Sciences” ( POCI-01-0145-FEDER-007274 ) and by project COMBINE (Santa Casa da Misericordia de Lisboa – Prémio Neurociências Mello e Castro). D.B. was supported by FCT PhD Programs ( PD/BD/105953/2014 ) and Programa Operacional Potencial Humano (POCH), in the scope of the BiotechHealth Program (Doctoral Program on Cellular and Molecular Biotechnology Applied to Health Sciences), Programa FLAD Helthcare 2020 and the project PARES (Prémio Albino Aroso). Eduardo Conde-Sousa was supported by a post-doctoral grant of the project PPBI-POCI-01-0145-FEDER-022122 , in the scope of Fundação para a Ciência e Tecnologia, Portugal (FCT) National Roadmap of Research Infrastructures. Appendix

Dhvar5- and MSI78-coated titanium are bactericidal against methicillin-resistant Staphylococcus aureus, immunomodulatory and osteogenic

This work was financed by Portuguese funds through FCT-Fundaç˜ao para a Ciˆencia e a Tecnologia/Minist´erio da Ciˆencia, Tecnologia e Inovaç˜ao in the framework of the grants/contracts for Bruna Costa (SFRH/BD/147,027/2019), Joana Coelho (2023.03848.BD), Fabíola Costa (CEECIND/01921/2017), Cl´audia Monteiro (art. 23 of Law no. 57/2016 & 57/2017) and M. Cristina L. Martins (LA/P/0070/2020). Nat´alia A. Costa thanks to Coordenaç˜ao de Aperfeiçoamento de Pessoal de Nível Superior - Brazil (CAPES) - Finance Codes 88887.600413/ 2021–00 and 88887.802752/2023–00, and S˜ao Paulo Research Foundation (FAPESP) (grants #2020/10125–9 and #2021/11461–5). M. Cristina L. Martins also acknowledges MOBILIsE Project, which has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement no. 951,723. XPS analysis was funded by the European Union’s H2020 project Sinfonia (N.857253).Infection is one of the major issues associated with the failure of orthopedic devices, mainly due to implant bacterial colonization, biofilm formation, and associated antibiotic resistance. Antimicrobial peptides (AMP) are a promising alternative to conventional antibiotics given their broad-spectrum of activity, low propensity to induce bacterial resistance, and ability to modulate host immune responses. Dhvar5 (LLLFLLKKRKKRKY) and MSI78 (GIGKFLKKAKKFGKAFVKILKK) are two AMP with broad-spectrum activity against bacteria, including methicillin-resistant Staphylococcus aureus (MRSA), one of the most problematic etiologic agents in Orthopedic Devices-Related Infections (ODRI). This work aims to evaluate the bactericidal, immunomodulatory and osteogenic potential of Dhvar5- and MSI78-coated titanium surfaces (AMP-Ti). Two AMP-Ti surfaces were successfully obtained by grafting Dhvar5 (0.8 ± 0.1 µM/mm2) or MSI78 (0.5 ± 0.3 µM/mm2) onto titanium substrates through a polydopamine layer. Both AMP-Ti were bactericidal against MRSA, eradicating bacteria upon contact for 6 h in a culture medium supplemented with human plasma proteins. The AMP-Ti immunomodulatory potential was evaluated using human primary macrophages, by assessing surfaces capacity to induce pro-/anti-inflammatory (M1/M2) markers and cytokines. While in naïve conditions both AMP-Ti surfaces slightly promoted the M2 marker CD163, in response to LPS and IFN-γ (simulating a bacterial infection), both AMP increased the M1 marker CCR7 and enhanced macrophage secretion of pro-inflammatory IL-6 and TNF-α cytokines, particularly for Ti-MSI78 surfaces. Additionally, both AMP-Ti surfaces were cytocompatible and promoted osteoblastic cell differentiation. This proof-of-concept study demonstrated the high potential of Dhvar5- and MSI78-Ti as antimicrobial coatings for ODRI prevention. STATEMENT OF SIGNIFICANCE: This study investigates the bactericidal effects of the antimicrobial peptides Dhvar5 and MSI78, immobilized on titanium (Ti) surfaces through a polydopamine coating, aiming at the prevention of Orthopedic-Device Related Infections (ODRIs). The developed coatings displayed MRSA-sterilizing activity, while revealing an immunomodulatory potential towards macrophages and promoting osteoblastic cell differentiation. This strategy allows a quick and easy immobilization of high quantities of AMP, unlike most other approaches, thus favoring its clinical translation

Dysthanasia: nursing professionals' perception

Dysthanasia means slow and painful death without quality of life. This study aimed to know whether nurses identify dysthanasia as part of the final process of the lives of terminal patients hospitalized at an adult ICU. This is an exploratory-qualitative study. Data were collected through semi-structured interviews with ten nurses with at least one year of experience in an ICU, and interpreted through content analysis. Results indicate that nurses understand and identify dysthanasia, do not agree with it and recognize elements of orthonasia as the adequate procedure for terminal patients. We conclude that nurses interpret dysthanasia as extending life with pain and suffering, while terminal patients are submitted to futile treatments that do not benefit them. They also identify dysthanasia using elements of orthonasia to explain it.Distanasia significa muerte lenta, con sufrimiento y sin calidad de vida. En esta investigación se buscó conocer si los enfermeros identifican la distanasia como parte del proceso final de la vida de personas en estado terminal, internadas en una UTI para adultos. El estudio es de naturaleza exploratoria, con abordaje cualitativo. Los datos fueron recolectados por medio de entrevista semiestructurada con 10 enfermeros con un mínimo de un año de experiencia en UTI; los datos fueron interpretados por el análisis de contenido. Se obtuvo como resultado que los enfermeros comprenden e identifican la distanasia y se oponen a la misma, presentando elementos de ortotanasia como procedimiento adecuado para pacientes en estado terminal. Se concluye que los enfermeros interpretan la distanasia como el prolongamiento de la vida con dolor y sufrimiento, en el cual los pacientes terminales son sometidos a tratamientos fútiles que no traen beneficios. También identifican la distanasia, usando elementos de la ortotanasia para hacerla explicita.Distanásia significa morte lenta, sofrida e sem qualidade de vida. Nesta pesquisa buscou-se conhecer se os enfermeiros identificam a distanásia como parte do processo final da vida de pessoas em terminalidade, internadas em UTI adulto. O estudo é de natureza exploratória, com abordagem qualitativa. Os dados foram coletados por meio de entrevista semiestruturada com 10 enfermeiros com, no mínimo, um ano de experiência em UTI, e interpretados pela análise de conteúdo. Teve-se como resultado que os enfermeiros compreendem e identificam a distanásia e se opõem à mesma, trazendo elementos da ortotanásia como procedimento adequado para pacientes em terminalidade. Conclui-se que os enfermeiros interpretam a distanásia como o prolongamento de vida com dor e sofrimento, onde os pacientes terminais são submetidos a tratamentos fúteis que não trazem benefícios. E também identificam a distanásia, usando elementos da ortotanásia para explicitá-la

EHA–EU MCL network guidelines for diagnosis and treatment of mantle cell lymphoma

Mantle cell lymphoma (MCL) is a relatively rare B‐cell lymphoma subtype, with a higher incidence among males and a median age of 70 years at diagnosis. MCL is characterized by clinically diverse behavior, from indolent disease to extremely aggressive, related to the presence of biological risk factors such as proliferation rate and TP53 mutations. Most often, patients present with disseminated disease, necessitating systemic treatment. Immunochemotherapy has historically been the mainstay of treatment, but recent data indicate that addition of novel agents, especially covalent Bruton tyrosine kinase inhibitors (cBTKi), may substantially improve outcome in younger and older patients, although a curative approach remains to be shown. In elderly patients, the standard of care is still immuno‐chemotherapy such as rituximab‐bendamustine, although this may be challenged by non‐chemotherapeutic options, such as rituximab plus cBTKi. For patients with relapsed or refractory disease, treatment options are developing rapidly, including CAR‐T cell therapy, novel BTK targeting agents, BCL2 inhibitors, and T‐cell engagers. In this clinical practice guideline, we present current evidence‐based recommendations for diagnosis, staging, treatment, and follow‐up of MCL

Identification and characterization of murine glycoprotein 2‐expressing intestinal dendritic cells

The intestinal lamina propria (LP) contains distinct subsets of classical dendritic cells (cDC), each playing key non-redundant roles in intestinal immune homeostasis. Here, we show that glycoprotein 2 (GP2), a GPI-anchored protein and receptor for bacterial type-I fimbriae, is selectively expressed by CD103+CD11b+ cDC in the murine small intestine (SI). GP2 expression was induced on CD103+CD11b+ cDC within the SI-LP and was regulated by IRF4, TGFβR1- and retinoic acid signalling. Mice selectively lacking Gp2 on CD103+CD11b+ cDC (huLang-Cre.gp2fl/fl mice) had normal numbers and proportions of innate and adaptive immune cells in the SI-LP suggesting that GP2 expression by CD103+CD11b+ cDC is not required for intestinal immune homoeostasis

Qualidade dos cuidados de enfermagem e satisfação do paciente atendido em um hospital de ensino

OBJECTIVES: assess the quality of nursing care, the patients' satisfaction and the correlation between both. METHOD: cross-sectional study, involving 275 patients hospitalized at a teaching hospital in the Central-West of Brazil. The data were collected through the simultaneous application of three instruments. Next, they were included in an electronic database and analyzed in function of the positivity, median value and Spearman's correlation coefficients. RESULTS: among the nursing care assessed, only two were considered safe - hygiene and physical comfort; nutrition and hydration - while the remainder were classified as poor. Nevertheless, the patients were satisfied with the care received in the domains assessed: technical-professional, confidence and educational. This can be justified by the weak to moderate correlation that was observed among these variables. CONCLUSION: Despite the quality deficit, the patients' satisfaction level with the nursing care received was high. These results indicate that the institution needs to center its objectives on a continuing evaluation system of the care quality, aiming to attend to the patients' expectations.OBJETIVOS: evaluar la calidad de los cuidados de enfermería, la satisfacción del paciente y la correlación entre ambos. MÉTODO: estudio trasversal con 275 pacientes internados en un hospital de enseñanza de la región Centro-Oeste de Brasil. Los datos fueron recolectados mediante la aplicación simultánea de tres instrumentos. A seguir, fueron digitalizados en un banco de datos electrónico y analizados en función de la positividad, valor de mediana y coeficientes de correlación de Spearman. RESULTADOS: entre los cuidados de enfermería evaluados, solamente dos fueron considerados seguros - higiene y conforto físico; nutrición e hidratación - y los demás clasificados como pobres. Sin embargo, los pacientes se mostraron satisfechos con los cuidados recibidos en los dominios evaluados: técnico-profesional, confianza y educacional. Eso puede ser justificado por la correlación débil a moderada observada entre esas variables. CONCLUSIÓN: A pesar del déficit de cualidad, fue encontrado alto nivel de satisfacción de los pacientes con los cuidados de enfermería recibidos. Tales resultados indican la necesidad de que la institución centre sus objetivos en un sistema de evaluación permanente de la cualidad del cuidado, visando atender a las expectativas de los pacientes.OBJETIVOS: avaliar a qualidade dos cuidados de enfermagem, a satisfação do paciente e a correlação entre ambos. MÉTODO: estudo transversal, realizado com 275 pacientes internados em um hospital de ensino da Região Centro-Oeste do Brasil. Os dados foram obtidos por meio da aplicação simultânea de três instrumentos. A seguir, foram digitados em banco de dados eletrônico e analisados em função da positividade, valor de mediana e coeficientes de correlação de Spearman. RESULTADOS: dentre os cuidados de enfermagem avaliados, apenas dois foram considerados seguros - higiene e conforto físico; nutrição e hidratação - e os demais foram classificados como pobres. Todavia, os pacientes mostraram-se satisfeitos com os cuidados recebidos nos domínios avaliados: técnico-profissional, confiança e educacional. Isso pode ser justificado pela correlação fraca a moderada observada entre essas variáveis. CONCLUSÃO: apesar do déficit de qualidade, houve alto nível de satisfação dos pacientes com os cuidados de enfermagem recebidos. Tais resultados apontam a necessidade de a instituição centrar seus objetivos num sistema de avaliação permanente da qualidade do cuidado, visando o atendimento das expectativas dos pacientes