Artificial Antigen Presenting Cells With Preclustered anti-CD28/-CD3/-LFA-1 Monoclonal Antibodies Are Highly Effective To Induce The Ex-Vivo Expansion Of Functional Human Antitumor T Cells

Effective adoptive T cell therapy requires the _ex vivo_ generation of functional T lymphocytes with a long lifespan _in vivo_. We evaluated _in vitro_ T cell expansion by artificial antigen presenting cells (aAPC) generated with activating (human anti-CD3), co-stimulating (human anti-CD28) and adhesion (human anti-LFA-1) monoclonal antibodies pre-clustered in microdomains (MDs) held by a liposome scaffold. The co-localization of T cell ligands in MDs and the targeting of an adhesion protein, increasing the efficiency of immunological synapse formations, represent the novelties of our system. These aAPCs allowed increased expansion of polyclonal CD4^+^ and CD8^+^ T cells and of tumor antigen-specific CD8^+^ T cells compared to anti-CD28- and anti-CD3-coated microbeads and to immobilized anti-CD3. These aAPCs allowed the generation of T cells displaying an immunophenotype consistent with long-term _in vivo_ persistence, without increasing the frequency of regulatory T cells. Finally, our aAPCs proved to be suitable for large scale T cell expansion required in immunotherapy trials

PRÁTICAS ASSISTENCIAIS DE ENFERMAGEM EM UM NÚCLEO DE OSTOMIA: RELATO DE EXPERIÊNCIA

Objective: Report an experience of residents’ nurses during its practices in a core of ostomates. Methods: This study it is an experience report describing aspects experienced by the authors in the period they developed specialized nursing care activities with ostomates and/or urinary incontinence patients. In this sense, such practices occurred in the Regional Center of the West of Paraná Ostomy, between July 28 and August 20, 2014. Results: They were attended 15 patients, 66.7 % were female. Age ranged between 35 and 68 years and 53.3 % of patients was from the host city core. With regard to the type of ostomy, it was found that 80% was colostomy patients. Moreover, there were nursing consultations covering the whole process, delivery of medical and hospital supplies and health education users. In addition, you could attend monthly meeting organized by members (patients). Conclusions: The practical specialized assistance developed by residents, in addition to strengthening scientific knowledge and personal values toward the service to ostomates patients, proved of great importance to the user who had the opportunity to raise subsidies to promoting self-care and improvement in their quality life.Objetivo: Reportar una experiencia de residentes enfermeras durante sus prácticas en un núcleo de ostomizados. Métodos: Este estudio es un relato de experiencia que describe aspectos experimentados por los autores en el período en que se desarrollaron las actividades de cuidado de enfermería especializados con ostomizados y/o pacientes de incontinencia urinaria. En este sentido, estas prácticas tuvieron lugar en el Centro Regional del Oeste de Paraná ostomía, entre el 28 de julio y 20 de agosto 2014. Resultados: Fueron atendidos 15 pacientes, 66,7 % eran mujeres. La edad osciló entre 35 y 68 años y el 53,3 % de los pacientes eran de la base de la ciudad anfitriona. En cuanto al tipo de ostomía, se encontró que el 80 % eran portador de colostomía. Además, no se Enfermería consultas teniendo en cuenta el proceso en su conjunto, la entrega de suministros médicos y hospitalarios y usuarios de educación sanitaria. Además, podríamos asistir a la reunión mensual organizada por los miembros (los pacientes). Conclusión: Asistencia especializada práctica desarrollada por los residentes, además de fortalecer el conocimiento científico y los valores personales en contra del servicio a los pacientes ostomizados, resultó de gran importancia para el usuario que tuvo la oportunidad de aumentar los subsidios para promover el autocuidado y la mejora de su calidad vida. Objetivo: Relatar a experiência vivenciada por enfermeiros residentes durante suas práticas assistenciais em um núcleo de ostomizados. Métodos: Este estudo trata-se de um relato de experiência que descreve aspectos vivenciados pelos autores no período em que desenvolveram atividades de assistência de enfermagem especializada junto a pacientes ostomizados e/ou com incontinência urinária. Nessa direção, as referidas práticas ocorreram no Núcleo Regional de Ostomizados do Oeste do Paraná, entre 28 de julho e 20 de agosto de 2014. Resultados: Foram atendidos 15 pacientes, sendo que 66,7% do gênero feminino. A idade variou entre 35 e 68 anos e 53,3% dos pacientes eram oriundos da cidade-sede do núcleo. No que se refere ao tipo de ostomia, verificou-se que 80% eram portador de colostomia. Outrossim, realizaram-se consultas de Enfermagem contemplando o processo como um todo, entrega de material médico-hospitalar e educação em saúde aos usuários. Além disso, pôde-se participar de encontro mensal promovido pelos associados (pacientes). Conclusões: As práticas assistências especializadas desenvolvidas pelos residentes, além de reforçar os conhecimentos científicos e valores pessoais frente ao atendimento aos pacientes ostomizados, mostraram-se de grande importância ao usuário o qual teve a oportunidade de angariar subsídios para promoção do autocuidado e melhoria em sua qualidade de vida

Inhibition of N-linked glycosylation impairs ALK phosphorylation and disrupts pro-survival signaling in neuroblastoma cell lines

<p>Abstract</p> <p>Background</p> <p>The Anaplastic Lymphoma Kinase (ALK) is an orphan receptor tyrosine kinase, which undergoes post-translational N-linked glycosylation. The catalytic domain of ALK was originally identified in the t(2;5) translocation that produces the unglycosylated oncogenic protein NPM-ALK, which occurs in Anaplastic Large Cell Lymphoma (ALCL). Recently, both germline and somatic activating missense mutations of ALK have been identified in neuroblastoma (NB), a pediatric cancer arising from neural crest cells. Moreover, we previously reported that ALK expression is significantly upregulated in advanced/metastatic NB. We hypothesized that ALK function may depend on N-linked glycosylation and that disruption of this post-translational modification would impair ALK activation, regardless the presence of either gene mutations or overexpression.</p> <p>Methods</p> <p>We employed tunicamycin to inhibit N-linked glycosylation. The following ALK-positive NB cell lines were used: SH-SY5Y and KELLY (ALK mutation F1174L), UKF-NB3 (ALK mutation R1275Q) and NB1 (ALK amplification). As a control, we used the NB cell lines LA1-5S and NB5 (no ALK expression), and the ALCL cell line SU-DHL1 (NPM-ALK).</p> <p>Results</p> <p>Tunicamycin treatment of ALK-positive NB cells resulted in a hypoglycosylated ALK band and in decreased amounts of mature full size receptor. Concomitantly, we observed a marked reduction of mature ALK phosphorylation. On the contrary, tunicamycin had no effects on NPM-ALK phosphorylation in SU-DHL1 cells. Moreover, phosphorylation levels of ALK downstream effectors (AKT, ERK1/2, STAT3) were clearly impaired only in ALK mutated/amplified NB cell lines, whereas no significant reduction was observed in both ALK-negative and NPM-ALK-positive cell lines. Furthermore, inhibition of N-linked glycosylation considerably impaired cell viability only of ALK mutated/amplified NB cells. Finally, the cleavage of the Poly-ADP-ribose-polymerase (PARP) suggested that apoptotic pathways may be involved in cell death.</p> <p>Conclusions</p> <p>In this study we showed that inhibition of N-linked glycosylation affects ALK phosphorylation and disrupts downstream pro-survival signaling, indicating that inhibition of this post-translational modification may be a promising therapeutic approach. However, as tunicamycin is not a likely candidate for clinical use other approaches to alter N-linked glycosylation need to be explored. Future studies will assess whether the efficacy in inhibiting ALK activity might be enhanced by the combination of ALK specific small molecule and N-linked glycosylation inhibitors.</p

Tumor de células granulares mimetizando uma úlcera eosinofílica

O tumor de células granulares (TCG) é uma neoplasia benigna incomum de tecido mole possível, decorrente de células neurais, especialmente células de Schwann. Na cavidade oral, o local mais frequente é a língua. Geralmente, a aparência clínica é um nódulo submucoso solitário, indolor e não ulcerado, localizado no dorso da língua. As mulheres têm duas vezes mais predileção do que os homens e a idade média ocorre entre a quarta e a sexta décadas. Este relato de caso consiste em um homem de 56 anos de idade, que referiu a evolução de 6 meses de uma lesão ulcerada na borda lateral direita da língua. Clinicamente, observou-se ulceração discóide com centro eritematoso elevado e halo branco. A hipótese clínica era uma úlcera eosinofílica. Após biópsia excisional e processos histológicos, um grupo predominante de células eosinofílicas com citoplasma granular elevado do tecido conjuntivo para a superfície ulcerada pôde ser visualizado. Uma reação imuno-histoquímica S-100 foi feita, e uma forte marcação dessas células confirmou o diagnóstico final do TCGGranular Cell Tumor (GCT) is an uncommon benign neoplasia of soft tissue possible arise from neural cells, specially Schwann cells. In oral cavity, the most frequent site is the tongue. Usually, the clinical appearance is a solitary submucosal nodule, painless and non-ulcerated, located in tongue dorsum. Female have twice most predilection than male and mean age occurs between fourth to six decades. This case report consists in a 56-years-old male referred the 6 moth evolution of an ulcerated lesion in the right lateral border of tongue. Clinically, was observed a discoidal ulceration with elevated erythematous center and whiteness halo. The clinical hypothesis was an eosinophilic ulcer. After excisional biopsy and histologic processes, a predominant group of eosinophilic cells with granular cytoplasm raised from connective tissue to ulcerated surface could be visualized. A S-100 immunohistochemical reaction was made, and a strong marking of these cells confirmed the final diagnosis of GC

Evaluation of Patient Satisfaction Using the EORTC IN-PATSAT32 Questionnaire and Surgical Outcome in Single-Port Surgery for Benign Adnexal Disease: Observational Comparison with Traditional Laparoscopy

Laparoscopic surgery has been demonstrated as a valid approach in almost all gynaecologic procedures including malignant diseases. Benefits of the minimally invasive approach over traditional open surgery have been well demonstrated in terms of minimal perioperative morbidity and reduced postoperative pain and hospital stay duration, with consequent quick postoperative recovery (Medeiros et al. (2009)). Single-port surgery resurfaced in gynaecology surgery in recent years and renewed interest among other surgeons and within the industry to develop this field (Podolsky et al. (2009)). Patient satisfaction is emerging as an increasingly important measure of quality which represents a complex entity that is dependent on patient demographics, comorbidities, disease, and, to a large extent, patient expectations (Tomlinson and Ko (2006)). It can be broadly thought to refer to all relevant experiences and processes associated with health care delivery (Jackson et al. (2001)). In this study we aim to compare single-port surgery (SPS) with conventional laparoscopy in terms of patient satisfaction using the EORTC IN-PATSAT32 questionnaire. We also evaluate the main surgical outcomes of both minimally invasive approaches

A Combined Approach Employing Chlorotoxin-Nanovectors and Low Dose Radiation To Reach Infiltrating Tumor Niches in Glioblastoma

Glioblastoma multiforme (GBM) is the most aggressive form of glioma, with life expectancy of around 2 years after diagnosis, due to recidivism and to the blood-brain barrier (BBB) limiting the amount of drugs which reach the residual malignant cells, thus contributing to the failure of chemotherapies. To bypass the obstacles imposed by the BBB, we investigated the use of nanotechnologies combined with radiotherapy, as a potential therapeutic strategy for GBM. We used poly­(lactic-<i>co</i>-glycolic acid) (PLGA) nanoparticles (PNP) conjugated to chlorotoxin (CTX), a peptide reported to bind selectively to glioma cells. Silver nanoparticles were entrapped inside the functionalized nanoparticles (Ag-PNP-CTX), to allow detection and quantification of the cellular uptake by confocal microscopy, both <i>in vitro</i> and <i>in vivo</i>. <i>In vitro</i> experiments performed with different human glioblastoma cell lines showed higher cytoplasmic uptake of Ag-PNP-CTX, with respect to nonfunctionalized nanoparticles. <i>In vivo</i> experiments showed that Ag-NP-CTX efficiently targets the tumor, but are scarcely effective in crossing the blood brain barrier in the healthy brain, where dispersed metastatic cells are present. We show here that single whole brain X-ray irradiation, performed 20 h before nanoparticle injection, enhances the expression of the CTX targets, MMP-2 and ClC-3, and, through BBB permeabilization, potently increases the amount of internalized Ag-PNP-CTX even in dispersed cells, and generated an efficient antitumor synergistic effect able to inhibit <i>in vivo</i> tumor growth. Notably, the application of Ag-PNP-CTX to irradiated tumor cells decreases the extracellular activity of MMP-2. By targeting dispersed GBM cells and reducing MMP-2 activity, the combined use of CTX-nanovectors with radiotherapy may represent a promising therapeutic approach toward GBM

Dual role of integrin Alpha-6 in glioblastoma: supporting stemness in proneural stem-like cells while inducing radioresistance in mesenchymal stem-like cells

Therapeutic resistance after multimodal therapy is the most relevant cause of glioblastoma (GBM) recurrence. Extensive cellular heterogeneity, mainly driven by the presence of GBM stem-like cells (GSCs), strongly correlates with patients' prognosis and limited response to therapies. Defining the mechanisms that drive stemness and control responsiveness to therapy in a GSC-specific manner is therefore essential. Here we investigated the role of integrin a6 (ITGA6) in controlling stemness and resistance to radiotherapy in proneural and mesenchymal GSCs subtypes. Using cell sorting, gene silencing, RNA-Seq, and in vitro assays, we verified that ITGA6 expression seems crucial for proliferation and stemness of proneural GSCs, while it appears not to be relevant in mesenchymal GSCs under basal conditions. However, when challenged with a fractionated protocol of radiation therapy, comparable to that used in the clinical setting, mesenchymal GSCs were dependent on integrin a6 for survival. Specifically, GSCs with reduced levels of ITGA6 displayed a clear reduction of DNA damage response and perturbation of cell cycle pathways. These data indicate that ITGA6 inhibition is able to overcome the radioresistance of mesenchymal GSCs, while it reduces proliferation and stemness in proneural GSCs. Therefore, integrin a6 controls crucial characteristics across GBM subtypes in GBM heterogeneous biology and thus may represent a promising target to improve patient outcomes.This work was supported by Agència Gestió Ajuts Universitaris i Recerca, Generalitat de Catalunya, grant number 2017SGR1014); Red Temática de investigación cooperativa en cáncer, grant number RD12/0036/0029); School of Nursing, grant number PREI-UB 17/005I; 18/010I; 19/009A; the Fondation ARC pour la recherche sur le cancer, the INSERM-CNRS ATIP-Avenir grant, the European Research Council (ERC) under the European Union’s Horizon 2020 (grant agreement no. 805225) and the NanoTheRad (Paris-Saclay University). High-throughput sequencing was performed by the ICGex platform of the Institut Curie supported by the grants ANR-10-EQPX-03, ANR-10-INBS-09-08, and INCa-DGOS-4654. Work of M.M. and L.P. is supported by Fondazione AIRC per la Ricerca sul Cancro (IG 18851). E.S. is presently supported by Fondazione Veronesi. A.E-C. is funded by ISCIII/MINECO (PT17/0009/0019) and co-funded by FEDER