Moving forward through consensus: protocol for a modified Delphi approach to determine the top research priorities in the field of orthopaedic oncology.

IntroductionOrthopaedic oncology researchers face several obstacles in the design and execution of randomised controlled trials, including finite fiscal resources to support the rising costs of clinical research and insufficient patient volume at individual sites. As a result, high-quality research to guide clinical practice has lagged behind other surgical subspecialties. A focused approach is imperative to design a research programme that is economical, streamlined and addresses clinically relevant endpoints. The primary objective of this study will be to use a consensus-based approach to identify research priorities for international clinical trials in orthopaedic oncology.Methods and analysisWe will conduct a 3-phase modified Delphi method consisting of 2 sequential rounds of anonymous web-based questionnaires (phases I and II), and an in-person consensus meeting (phase III). Participants will suggest research questions that they believe are of particular importance to the field (phase I), and individually rate each proposed question on 5 criteria (phase II). Research questions that meet predetermined consensus thresholds will be brought forward to the consensus meeting (phase III) for discussion by an expert panel. Following these discussions, the expert panel will be asked to assign scores for each research question, and research questions meeting predetermined criteria will be brought forward for final ranking. The expert panel will then be asked to rank the top 3 research questions, and these 3 research questions will be distributed to the initial group of participants for validation.Ethics and disseminationAn ethics application is currently under review with the Hamilton Integrated Research Ethics Board in Hamilton, Ontario, Canada. The results of this initiative will be disseminated through peer-reviewed publications and conference presentations

The Oesophageal Squamous Cell Carcinoma Cell Line COLO-680N Fails to Support Sustained Cryptosporidium parvum Proliferation

Cryptosporidium parvum is an important diarrhoea-associated protozoan, which is difficult to propagate in vitro. In 2017, a report described a continuous culture of C. parvum Moredun strain, in the oesophageal squamous cell carcinoma cell line COLO-680N, as an easy-to-use system for C. parvum propagation and continuous production of oocysts. Here, we report that—using the Köllitsch strain of C. parvum—even though COLO-680N cells, indeed, allowed parasite invasion and early asexual parasite replication, C. parvum proliferation decreased after the second day post infection. Considering recurring studies, reporting on successful production of newly generated Cryptosporidium oocysts in the past, and the subsequent replication failure by other research groups, the current data stand as a reminder of the importance of reproducibility of in vitro systems in cryptosporidiosis research. This is of special importance since it will only be possible to develop promising strategies to fight cryptosporidiosis and its ominous consequences for both human and animal health by a continuous and reliable methodological progress

First Metabolic Insights into Ex Vivo Cryptosporidium parvum-Infected Bovine Small Intestinal Explants Studied under Physioxic Conditions

The apicomplexan Cryptosporidium parvum causes thousands of human deaths yearly. Since bovines represent the most important reservoir of C. parvum, the analysis of infected bovine small intestinal (BSI) explants cultured under physioxia offers a realistic model to study C. parvum–host cell–microbiome interactions. Here, C. parvum-infected BSI explants and primary bovine small intestinal epithelial cells were analysed for parasite development and metabolic reactions. Metabolic conversion rates in supernatants of BSI explants were measured after infection, documenting an immediate parasite-driven metabolic interference. Given that oxygen concentrations affect cellular metabolism, measurements were performed at both 5% O2 (physiological intestinal conditions) and 21% O2 (commonly used, hyperoxic lab conditions). Overall, analyses of C. parvum-infected BSI explants revealed a downregulation of conversion rates of key metabolites—such as glucose, lactate, pyruvate, alanine, and aspartate—at 3 hpi, followed by a rapid increase in the same conversion rates at 6 hpi. Moreover, PCA revealed physioxia as a driving factor of metabolic responses in C. parvum-infected BSI explants. Overall, the ex vivo model described here may allow scientists to address pending questions as to how host cell–microbiome alliances influence intestinal epithelial integrity and support the development of protective intestinal immune reactions against C. parvum infections in a realistic scenario under physioxic conditions

Fricción interna y comportamiento anelástico en sólidos

Se presentan los conceptos de Fricción Interna (FI), anelasticidad y espectroscopia mecánica, como una técnica potente para la determinación de las propiedades mecánicas dinámicas en sólidos. Se comentan algunos de los logros de la técnica, fundamentalmente en el campo de la física del estado sólido

Metabolic Signatures of Cryptosporidium parvum-Infected HCT-8 Cells and Impact of Selected Metabolic Inhibitors on C. parvum Infection under Physioxia and Hyperoxia

Cryptosporidium parvum is an apicomplexan zoonotic parasite recognized as the second leading-cause of diarrhoea-induced mortality in children. In contrast to other apicomplexans, C. parvum has minimalistic metabolic capacities which are almost exclusively based on glycolysis. Consequently, C. parvum is highly dependent on its host cell metabolism. In vivo (within the intestine) infected epithelial host cells are typically exposed to low oxygen pressure (1–11% O2, termed physioxia). Here, we comparatively analyzed the metabolic signatures of C. parvum-infected HCT-8 cells cultured under both, hyperoxia (21% O2), representing the standard oxygen condition used in most experimental settings, and physioxia (5% O2), to be closer to the in vivo situation. The most pronounced effect of C. parvum infection on host cell metabolism was, on one side, an increase in glucose and glutamine uptake, and on the other side, an increase in lactate release. When cultured in a glutamine-deficient medium, C. parvum infection led to a massive increase in glucose consumption and lactate production. Together, these results point to the important role of both glycolysis and glutaminolysis during C. parvum intracellular replication. Referring to obtained metabolic signatures, we targeted glycolysis as well as glutaminolysis in C. parvum-infected host cells by using the inhibitors lonidamine [inhibitor of hexokinase, mitochondrial carrier protein (MCP) and monocarboxylate transporters (MCT) 1, 2, 4], galloflavin (lactate dehydrogenase inhibitor), syrosingopine (MCT1- and MCT4 inhibitor) and compound 968 (glutaminase inhibitor) under hyperoxic and physioxic conditions. In line with metabolic signatures, all inhibitors significantly reduced parasite replication under both oxygen conditions, thereby proving both energy-related metabolic pathways, glycolysis and glutaminolysis, but also lactate export mechanisms via MCTs as pivotal for C. parvum under in vivo physioxic conditions of mammals

Clinical utility of intralesional methotrexate to distinguish crateriform keratinocytic tumors before surgery

Clinical utility of intralesional methotrexate to distinguish crateriform keratinocytic tumors before surgeryDear Editors,Keratoacanthoma (KA) and cutaneous squamous cell carci-noma (CSCC) may adopt an identical crateriform morpho-logy. Nowadays, the debate about whether KA is a distinct entity, or a low-grade variant of cutaneous squamous cell carcinoma (CSCC) still persists. Since CSCC is a more ag-gressive neoplasm, misdiagnosing crateriform lesions may have a negative impact on the patient's prognosis. Evaluating a partial biopsy is extremely challenging to confidently dis-tinguish KA from CSCC [1]. No distinctive gene expression profiles have been identified and no pathognomonic criteria to unequivocally differentiate between KA and CSCC exist [2]. Consequently, the surgical approach remains the gold standard in the management of crateriform tumors, especi-ally those arising on the face

Validación de un nuevo método de dosificación de metadona en pacientes del Programa de Mantenimiento con Metadona

Validació

Ocean Circulation over Formigas and Ormonde Seamounts

Seamounts constitute an obstacle to the free ocean flow, modifying the patter of circulation. As a result of these alterations, a variety of hydrodynamical processes and phenomena may take place in seamounts, among others, Taylor columns/caps. These oceanographic effects may turn seamounts into very productive ecosystems with high biodiversity. Under these conditions seamounts provide a particularly good environment for the settle of some organisms, acting as stepping stones and contributing to its dispersal. In this study, we verify if these oceanographic effects explain the presence of cold-water corals of Mediterranean origin in the Atlantic. To achieve this, three seamounts in the path of the Mediterranean Outflow Water (MOW) through the Eastern North Atlantic were selected: the Gazul mud volcano, and the Ormonde and Formigas seamounts. In order to determine the hydrographic and dynamical conditions in each one of the three locations, CTD, LADPC and biochemical observations were carried out. Taylor columns were not observed in any of the three sampled areas. Although we found suggestions of upwelling/downwelling systems, their effect was barely noticed in the circulation pattern. The oceanographic processes in those areas are more influenced by the vertical distribution of water masses, which determine the stability of the water column. Moreover, the high values of the Brunt-Väisälä frequency around the MOW halocline can lead to the formation of internal waves. These perturbations in the water column can enhance the vertical mixing, producing suspension, which, in turn, could affect the vertical distribution of cold-water corals