Perceptions and experiences of onco-haematology professionals in relation to advance care planning: a qualitative study.

PURPOSE: The objective of this study is to identify the beliefs, values, perceptions, and experiences of medical oncology, radiation oncology, and clinical haematology professionals about the advance care planning process. METHODS: Qualitative exploratory study. There were four focus groups with 14 nurses and 12 physicians (eight medical oncology, one radiation oncology, three haematology). A reflexive thematic analysis of the data obtained was performed. RESULTS: We identified 20 thematic categories, which we grouped into four themes: lack of knowledge about advance care planning; perception of the advance care planning process: knowledge acquired from practice; barriers and facilitators for the implementation of advance care planning; and communication as a key aspect of advance care planning. CONCLUSIONS: The participants valued advance care planning as an early intervention tool that promotes autonomy. They perceived difficulties in approaching planning due to lack of knowledge, training, and time. They identified the therapeutic relationship with the person, the participation of the person's loved ones, teamwork, and communication skills as essential to ensuring the quality of the process. Finally, they recognised that palliative care professionals provide added value in supporting planning processes.

Targeting of early endosomes by autophagy facilitates EGFR recycling and signalling.

Despite recently uncovered connections between autophagy and the endocytic pathway, the role of autophagy in regulating endosomal function remains incompletely understood. Here, we find that the ablation of autophagy-essential players disrupts EGF-induced endocytic trafficking of EGFR. Cells lacking ATG7 or ATG16L1 exhibit increased levels of phosphatidylinositol-3-phosphate (PI(3)P), a key determinant of early endosome maturation. Increased PI(3)P levels are associated with an accumulation of EEA1-positive endosomes where EGFR trafficking is stalled. Aberrant early endosomes are recognised by the autophagy machinery in a TBK1- and Gal8-dependent manner and are delivered to LAMP2-positive lysosomes. Preventing this homeostatic regulation of early endosomes by autophagy reduces EGFR recycling to the plasma membrane and compromises downstream signalling and cell survival. Our findings uncover a novel role for the autophagy machinery in maintaining early endosome function and growth factor sensing.

Update on Epithelial-Mesenchymal Plasticity in Cancer Progression.

Epithelial-mesenchymal transition (EMT) is a cellular process by which epithelial cells lose their characteristics and acquire mesenchymal traits to promote cell movement. This program is aberrantly activated in human cancers and endows tumor cells with increased abilities in tumor initiation, cell migration, invasion, metastasis, and therapy resistance. The EMT program in tumors is rarely binary and often leads to a series of gradual or intermediate epithelial-mesenchymal states. Functionally, epithelial-mesenchymal plasticity (EMP) improves the fitness of cancer cells during tumor progression and in response to therapies. Here, we discuss the most recent advances in our understanding of the diverse roles of EMP in tumor initiation, progression, metastasis, and therapy resistance and address major clinical challenges due to EMP-driven phenotypic heterogeneity in cancer. Uncovering novel molecular markers and key regulators of EMP in cancer will aid the development of new therapeutic strategies to prevent cancer recurrence and overcome therapy resistance.

[Food cravings and aversions in pregnant women in the Buenos Aires Conurban, Argentina]. / Antojos y aversiones alimentarias en gestantes del Conurbano Bonaerense, Argentina.

INTRODUCTION: Pregnant women often experience food aversions and cravings, of which little is known about their characteristics and consequences. The objective was to know the prevalence of food cravings and aversions, the characteristics of the pattern of foods that are craved or avoided, and the reasons behind their presence. METHODS: Observational, descriptive and cross-sectional study. A validated questionnaire was applied to 370 pregnant and postpartum women in public hospitals. Maternal and neonatal variables were measured with descriptive statistics and those associated with the phenomena under study were identified using logistic regression models and cluster analysis using the multivariate technique. RESULTS: A presence of cravings from 71 to 80% and aversions from 55 to 65% was detected. The most craved foods were fruits and sweet foods and the most avoided meats and mate. The maternal characteristics predictive of cravings were: being younger (OR 0.94), vomiting (OR 2.23), and having gained more weight than expected were negatively associated with the presence of cravings (OR 0.44). The variables associated with the aversions were the absence of a history of hypertension (OR 0.13), a history of macrosomia (OR 2.70), nausea (OR 1.86) and complications during pregnancy (OR 2.23). DISCUSSION: This work allowed to characterize food cravings and aversions during pregnancy and to know their high frequency.

Introducción: Las gestantes suelen experimentar aversiones y antojos alimentarios, de los cuales se conocen poco sus características y consecuencias. El objetivo fue conocer la prevalencia de los antojos y aversiones alimentarias, las características del patrón de alimentos que son antojados o evitados y los motivos que subyacen ante su presencia. Métodos: Estudio observacional, descriptivo y transversal. Un cuestionario validado se aplicó a 370 embarazadas y puérperas en hospitales públicos. Las variables maternas y neonatales se midieron con estadísticas descriptivas y se identificaron aquellas asociadas a los fenómenos en estudio empleando modelos de regresión logística y un análisis por clusters utilizando la técnica multivariante. Resultados: Se detectó una presencia de antojos del 71 al 80% y de aversiones del 55 al 65%. Los alimentos más antojados fueron las frutas y los alimentos dulces y los mayormente rechazados las carnes y el mate. Las características maternas predictoras de antojos fueron: tener menor edad (OR 0.94), vómitos (OR 2.23), y el haber ganado más peso de lo esperado se asoció negativamente con la presencia de antojos (OR 0.44). Las variables asociadas a las aversiones fueron la ausencia de antecedentes de hipertensión (OR 0.13), antecedentes de macrosomía (OR 2.70), náuseas (OR 1.86) y complicaciones durante el embarazo (OR 2.23). Discusión: Este trabajó permitió caracterizar los antojos y aversiones alimentarias durante el embarazo y conocer su elevada frecuencia.

Fibroblast activation protein drives tumor metastasis via a protease-independent role in invadopodia stabilization.

During metastasis, tumor cells invade through the basement membrane and intravasate into blood vessels and then extravasate into distant organs to establish metastases. Here, we report a critical role of a transmembrane serine protease fibroblast activation protein (FAP) in tumor metastasis. Expression of FAP and TWIST1, a metastasis driver, is significantly correlated in several types of human carcinomas, and FAP is required for TWIST1-induced breast cancer metastasis to the lung. Mechanistically, FAP is localized at invadopodia and required for invadopodia-mediated extracellular matrix degradation independent of its proteolytic activity. Live cell imaging shows that association of invadopodia precursors with FAP at the cell membrane promotes the stabilization and growth of invadopodia precursors into mature invadopodia. Together, our study identified FAP as a functional target of TWIST1 in driving tumor metastasis via promoting invadopodia-mediated matrix degradation and uncovered a proteolytic activity-independent role of FAP in stabilizing invadopodia precursors for maturation.

Mutation of the Conserved Threonine 8 within the Human ARF Tumour Suppressor Protein Regulates Autophagy.

BACKGROUND: The ARF tumour suppressor plays a well-established role as a tumour suppressor, halting cell growth by both p53-dependent and independent pathways in several cellular stress response circuits. However, data collected in recent years challenged the traditional role of this protein as a tumour suppressor. Cancer cells expressing high ARF levels showed that its expression, far from being dispensable, is required to guarantee tumour cell survival. In particular, ARF can promote autophagy, a self-digestion pathway that helps cells cope with stressful growth conditions arising during both physiological and pathological processes. METHODS: We previously showed that ARF is regulated through the activation of the protein kinase C (PKC)-dependent pathway and that an ARF phospho-mimetic mutant on the threonine residue 8, ARF-T8D, sustains cell proliferation in HeLa cells. We now explored the role of ARF phosphorylation in both basal and starvation-induced autophagy by analysing autophagic flux in cells transfected with either WT and ARF phosphorylation mutants by immunoblot and immunofluorescence. RESULTS: Here, we show that endogenous ARF expression in HeLa cells is required for starvation-induced autophagy. Further, we provide evidence that the hyper-expression of ARF-T8D appears to inhibit autophagy in both HeLa and lung cancer cells H1299. This effect is due to the cells' inability to elicit autophagosomes formation upon T8D expression. CONCLUSIONS: Our results lead to the hypothesis that ARF phosphorylation could be a mechanism through which the protein promotes or counteracts autophagy. Several observations underline how autophagy could serve a dual role in cancer progression, either protecting healthy cells from damage or aiding cancerous cells to survive. Our results indicate that ARF phosphorylation controls protein's ability to promote or counteract autophagy, providing evidence of the dual role played by ARF in cancer progression.

Matrix Degradation Assay to Measure the Ability of Tumor Cells to Degrade Extracellular Matrix.

During the metastatic process, carcinoma cells form invadopodia, F-actin enriched protrusive structures, to degrade the extracellular matrix (ECM) in order to invade the surrounding stroma and intravasate into the circulatory system. In this chapter, we describe the 2D-fluorescent matrix degradation assay, a highly sensitive and reproducible in vitro method used to measure invadopodia-mediated ECM degradation. We provide a detailed protocol on how to prepare the glass coverslips with fluorescent gelatin matrix and a standardized method to quantify gelatin degradation and invadopodia formation in order to evaluate cell invasion.

Dual Role of the Alternative Reading Frame ARF Protein in Cancer.

The CDKN2a/ARF locus expresses two partially overlapping transcripts that encode two distinct proteins, namely p14ARF (p19Arf in mouse) and p16INK4a, which present no sequence identity. Initial data obtained in mice showed that both proteins are potent tumor suppressors. In line with a tumor-suppressive role, ARF-deficient mice develop lymphomas, sarcomas, and adenocarcinomas, with a median survival rate of one year of age. In humans, the importance of ARF inactivation in cancer is less clear whereas a more obvious role has been documented for p16INK4a. Indeed, many alterations in human tumors result in the elimination of the entire locus, while the majority of point mutations affect p16INK4a. Nevertheless, specific mutations of p14ARF have been described in different types of human cancers such as colorectal and gastric carcinomas, melanoma and glioblastoma. The activity of the tumor suppressor ARF has been shown to rely on both p53-dependent and independent functions. However, novel data collected in the last years has challenged the traditional and established role of this protein as a tumor suppressor. In particular, tumors retaining ARF expression evolve to metastatic and invasive phenotypes and in humans are associated with a poor prognosis. In this review, the recent evidence and the molecular mechanisms of a novel role played by ARF will be presented and discussed, both in pathological and physiological contexts.

[Is the legally responsible party indeed responsible? An ethical-legal question on the term]. / O responsável legal é de fato o responsável? Um questionamento ético-legal sobre o termo.

The authors discuss the legal and ethical meaning of the expression "Third-Party Consent" by questioning its limits. It is indeed shown that it does not satisfactorily meet what is called third-party consent because this would require legal endorsement by legal codes and norms which, in fact does not occur. As such, the expression "third-party consent", whenever used, may not provide the professional with the normative, ethical and legal support needed for professional performance.

Dynamics of p14ARF and Focal Adhesion Kinase-Mediated Autophagy in Cancer.

It has been widely shown that the focal adhesion kinase (FAK) is involved in nearly every aspect of cancer, from invasion to metastasis to epithelial⁻mesenchymal transition and maintenance of cancer stem cells. FAK has been shown to interact with p14ARF (alternative reading frame)—a well-established tumor suppressor—and functions in the negative regulation of cancer through both p53-dependent and -independent pathways. Interestingly, both FAK and ARF (human and mouse counterpart) proteins, as well as p53, are involved in autophagy—a process of “self-digestion”—whose main function is the recycling of cellular components and quality control of proteins and organelles. In the last years, an unexpected role of p14ARF in the survival of cancer cells has been underlined in different cellular contexts, suggesting a novel pro-oncogenic function of this protein. In this review, the mechanisms whereby ARF and FAK control autophagy are presented, as well as the role of autophagy in cell migration and spreading. Integrated investigation of these cell functions is extremely important to understand the mechanism of the basis of cell transformation and migration and thus cancer development.

PKC Dependent p14ARF Phosphorylation on Threonine 8 Drives Cell Proliferation.

ARF role as tumor suppressor has been challenged in the last years by several findings of different groups ultimately showing that its functions can be strictly context dependent. We previously showed that ARF loss in HeLa cells induces spreading defects, evident as rounded morphology of depleted cells, accompanied by a decrease of phosphorylated Focal Adhesion Kinase (FAK) protein levels and anoikis. These data, together with previous finding that a PKC dependent signalling pathway can lead to ARF stabilization, led us to the hypothesis that ARF functions in cell proliferation might be regulated by phosphorylation. In line with this, we show here that upon spreading ARF is induced through PKC activation. A constitutive-phosphorylated ARF mutant on the conserved threonine 8 (T8D) is able to mediate both cell spreading and FAK activation. Finally, ARF-T8D expression confers growth advantage to cells thus leading to the intriguing hypothesis that ARF phosphorylation could be a mechanism through which pro-proliferative or anti proliferative signals could be transduced inside the cells in both physiological and pathological conditions.

Antojos y aversiones alimentarias en gestantes del Conurbano Bonaerense, Argentina / Food cravings and aversions in pregnant women in the Buenos Aires Conurban, Argentina

Resumen Introducción : Las gestantes suelen experimentar aversiones y antojos alimentarios, de los cuales se cono cen poco sus características y consecuencias. El objetivo fue conocer la prevalencia de los antojos y aversiones alimentarias, las características del patrón de alimentos que son antojados o evitados y los motivos que subyacen ante su presencia. Métodos : Estudio observacional, descriptivo y trans versal. Un cuestionario validado se aplicó a 370 embara zadas y puérperas en hospitales públicos. Las variables maternas y neonatales se midieron con estadísticas descriptivas y se identificaron aquellas asociadas a los fenómenos en estudio empleando modelos de regresión logística y un análisis por clusters utilizando la técnica multivariante. Resultados : Se detectó una presencia de antojos del 71 al 80% y de aversiones del 55 al 65%. Los alimentos más antojados fueron las frutas y los alimentos dulces y los mayormente rechazados las carnes y el mate. Las características maternas predictoras de antojos fueron: tener menor edad (OR 0.94), vómitos (OR 2.23), y el haber ganado más peso de lo esperado se asoció ne gativamente con la presencia de antojos (OR 0.44). Las variables asociadas a las aversiones fueron la ausencia de antecedentes de hipertensión (OR 0.13), antecedentes de macrosomía (OR 2.70), náuseas (OR 1.86) y complica ciones durante el embarazo (OR 2.23). Discusión : Este trabajó permitió caracterizar los an tojos y aversiones alimentarias durante el embarazo y conocer su elevada frecuencia.

Abstract Introduction : Pregnant women often experience food aversions and cravings, of which little is known about their characteristics and consequences. The objective was to know the prevalence of food cravings and aver sions, the characteristics of the pattern of foods that are craved or avoided, and the reasons behind their presence. Methods : Observational, descriptive and cross-sec tional study. A validated questionnaire was applied to 370 pregnant and postpartum women in public hospi tals. Maternal and neonatal variables were measured with descriptive statistics and those associated with the phenomena under study were identified using logistic regression models and cluster analysis using the mul tivariate technique. Results : A presence of cravings from 71 to 80% and aversions from 55 to 65% was detected. The most craved foods were fruits and sweet foods and the most avoided meats and mate. The maternal characteristics predictive of cravings were: being younger (OR 0.94), vomiting (OR 2.23), and having gained more weight than expected were negatively associated with the presence of cravings (OR 0.44). The variables associated with the aversions were the absence of a history of hypertension (OR 0.13), a history of macrosomia (OR 2.70), nausea (OR 1.86) and complications during pregnancy (OR 2.23). Discussion : This work allowed to characterize food cravings and aversions during pregnancy and to know their high frequency.

Plasma modified PLA electrospun membranes for actinorhodin production intensification in Streptomyces coelicolor immobilized-cell cultivations.

Most of industrially relevant bioproducts are produced by submerged cultivations of actinomycetes. The immobilization of these Gram-positive filamentous bacteria on suitable porous supports may prevent mycelial cell-cell aggregation and pellet formation which usually negatively affect actinomycete submerged cultivations, thus, resulting in an improved biosynthetic capability. In this work, electrospun polylactic acid (PLA) membranes, subjected or not to O2-plasma treatment (PLA-plasma), were used as support for immobilized-cell submerged cultivations of Streptomyces coelicolor M145. This strain produces different bioactive compounds, including the blue-pigmented actinorhodin (ACT) and red-pigmented undecylprodigiosin (RED), and constitutes a model for the study of antibiotic-producing actinomycetes. Wet contact angles and X-ray photoelectron spectroscopy analysis confirmed the increased wettability of PLA-plasma due to the formation of polar functional groups such as carboxyl and hydroxyl moieties. Scanning electron microscope observations, carried out at different incubation times, revealed that S. coelicolor immobilized-cells created a dense "biofilm-like" mycelial network on both kinds of PLA membranes. Cultures of S. coelicolor immobilized-cells on PLA or PLA-plasma membranes produced higher biomass (between 1.5 and 2 fold) as well as higher levels of RED and ACT than planktonic cultures. In particular, cultures of immobilized-cells on PLA and PLA-plasma produced comparable levels of RED that were approximatively 4 and 5 fold higher than those produced by planktonic cultures, respectively. In contrast, levels of ACT produced by immobilized-cell cultures on PLA and PLA-plasma were different, being 5 and 10 fold higher than those of planktonic cultures, respectively. Therefore, this is study demonstrated the positive influence of PLA membrane on growth and secondary metabolite production in S. coelicolor and also revealed that O2-plasma treated PLA membranes specifically promoted higher ACT production than not treated membranes.

Effect of PCL/PEG-Based Membranes on Actinorhodin Production in Streptomyces coelicolor Cultivations.

The actinomycetes, Gram-positive filamentous bacteria, are the most prolific source of natural occurring antibiotics. At an industrial level, antibiotics from actinomycete strains are produced by means of submerged fermentations, where one of the major factors negatively affecting bioproductivity is the pellet-shaped biomass growth. The immobilization of microorganisms on properly chosen supports prevents cell-cell aggregation resulting in improving the biosynthetic capability. Thus, novel porous biopolymer-based devices are developed by combining melt mixing and particulate leaching. In particular, polycaprolactone (PCL), polyethylene glycol (PEG), and sodium chloride (NaCl) with different grain sizes are used to prepare PCL/PEG/NaCl blends in the melt. These blends are then leached to obtain PCL-based porous membranes that are used as solid support for the growth of Streptomyces coelicolor, a model streptomycete used to produce various antibiotics including the blue colored actinorhodin (ACT). Thereafter, the effect of the devices' characteristics on the bacterial growth and on the production ACT is evaluated. The results showed that ACT production is strongly dependent on the pore size distribution of the device. Moreover, membranes with pores ranging from 90 to 110 µm are able to offer a potential improvement in volumetric productivity of ACT if compared to conventional submerged liquid culture.

O responsável legal é de fato o responsável? Um questionamento ético-legal sobre o termo / Is the legally responsible party indeed responsible?An ethical-legal question on the term

Os autores discutem o significado ético e jurídico da expressão "responsável legal", questionando seus limites. Demonstram que de fato ela não atende satisfatoriamente o que se denomina responsabilidade legal, pois para tanto teria que encontrar amparo nos códigos e normas legais, o que, de fato, não acontece. Assim, a expressão representante legal pode não possibilitar ao profissional, quando de sua utilização respaldo ético e legal normativo a sua atividade profissional.

The authors discuss the legal and ethical meaning of the expression "Third-Party Consent" by questioning its limits. It is indeed shown that it does not satisfactorily meet what is called third-party consent because this would require legal endorsement by legal codes and norms which, in fact does not occur. As such, the expression "third-party consent", whenever used, may not provide the professional with the normative, ethical and legal support needed for professional performance.

Informe final del grupo de estudio de enfermedad celíaca / Final report of the group of celiac disease

Elección del tema de trabajo: Enfermedad Celíaca. Adherencia a la Dieta Libre de Gluten. Revisión bibliográfica sobre el tema: Diferentes formas de medir la Adherencia a la Dieta Libre de Gluten. Revisiones y artículos de los últimos 10 años y trabajos de mayor antig³edad referentes en el tema. Los objetivos son: 1. Resumir el conocimiento existente sobre la definición de la adherencia terapéutica. 2. Identificar qué entiende la comunidad científica por adherencia y definir la magnitud del problema.3. Listar las diferentes formas de medir adherencia y si cuentan con validez científica. 4. Promover el debate de los temas relacionados con la adherencia terapéutica.(AU)

Informe final del grupo de estudio de enfermedad celíaca / Final report of the group of celiac disease

Elección del tema de trabajo: Enfermedad Celíaca. Adherencia a la Dieta Libre de Gluten. Revisión bibliográfica sobre el tema: Diferentes formas de medir la Adherencia a la Dieta Libre de Gluten. Revisiones y artículos de los últimos 10 años y trabajos de mayor antigüedad referentes en el tema. Los objetivos son: 1. Resumir el conocimiento existente sobre la definición de la adherencia terapéutica. 2. Identificar qué entiende la comunidad científica por adherencia y definir la magnitud del problema.3. Listar las diferentes formas de medir adherencia y si cuentan con validez científica. 4. Promover el debate de los temas relacionados con la adherencia terapéutica.

Cirugía endoscópica transluminal NOTES: resultados experimentales iniciales / Natural orifice transluminal endoscopic surgery (NOTES): initial experimental results

Introducción: La aparición de la cirugía endoscópica transorificial (NOTES) y de nuevos prototipos para su realización nos llevó a diseñar un programa de cirugía experimental. Material y métodos: Se ha realizado durante 1 año cirugía de NOTES en cerdas, cumpliendo la normativa europea de experimentación animal. Se han practicado 90 intervenciones sin supervivencia animal. En este estudio se ha evaluado: a) complejidad de las vías de acceso (transgástrica, transvaginal, transesofágica y transumbilical); b) técnicas de ayuda para la orientación temporoespacial, y c) posibilidades técnicas de cierre de los orificios viscerales (clips, t-bars®, clips de Obesco® y sutura por endoscopia). Las intervenciones practicadas mediante uno o dos endoscopios han sido: anexectomía, colecistectomía, biopsia de adenopatías mediastínicas, pancreatectomía distal y gastroyeyunostomía. Resultados: Esta experiencia nos ha permitido conocer una serie de aspectos técnicos imprescindibles para el desarrollo del NOTES: a) el neumoperitoneo con CO2 es más seguro para la entrada; b) el tamaño del orificio es importante para limitar la contaminación; c) la entrada con punción y dilatación con guía es más segura; d) la buena localización de la salida gástrica facilita el abordaje de las vísceras a disecar; e) la hemorragia intrabdominal es de difícil control; f) el cierre con garantías absolutas de fuga no existe, los clips no lo garantizan, los t-bars® y clips de Obesco® y la sutura endoscópica pueden funcionar, y g) la posibilidad de utilizar endoscopios que triangulan facilita la disección. Conclusiones: La NOTES requiere de un equipo multidisciplinario con cirujanos laparoscopistas y endoscopistas. La realización de NOTES pura es compleja; posiblemente las formas híbridas o la vía transumbilical sean fases intermedias (AU)

Introduction: The advent of natural orifice endoscopic surgery (NOTES) and new prototypes for performing this surgical procedure led us to design an experimental animal surgical programme. Material and method: NOTES was performed over a period of one year, in sows, following the European guidelines on the use of experimental animals. Ninety operations were performed with no animals surviving. The following aspects were assessed: 1. Access route complexity (transgastric, transvaginal, transesophageal and transumbilical). 2. Support measures for temporal/spatial orientation. 3. Technical possibilities for visceral orifice closure (clips, T-bars®, Obesco® clips and endoscopic suture). Resections of fallopian tubes, ovaries, gallbladder, mediastinal lymph nodes, tail of the pancreas and gastrojejunal derivations were performed with one or two endoscopes. Results: This experience enabled us to highlight a series of technical aspects essential for these techniques: 1. Pneumoperitoneum with CO2 is safer for entry. 2. Orifice size is important to limit contamination. 3. Puncture entry and guided dilation is safer. 4. Good gastric exit location makes it easier to approach viscera to be resected. 5. Intra-abdominal haemorrhage is difficult to control. 6. Leak-free closure cannot be guaranteed with clips, but t-bars® and Obesco® clips may be effective. 7. Endoscopes that permit triangulation may facilitate the dissection and endoscopic suture. Conclusions: NOTES requires a multidisciplinary team comprising laparoscopic and endoscopic surgeons. Pure NOTES is complex and hybrid forms or transumbilical route could be intermediate steps (AU)