A Temperature-Jump Optical Trap for Single-Molecule Manipulation

[EN] To our knowledge, we have developed a novel temperature-jump optical tweezers setup that changes the temperature locally and rapidly. It uses a heating laser with a wavelength that is highly absorbed by water so it can cover a broad range of temperatures. This instrument can record several force-distance curves for one individual molecule at various temper- atures with good thermal and mechanical stability. Our design has features to reduce convection and baseline shifts, which have troubled previous heating-laser instruments. As proof of accuracy, we used the instrument to carry out DNA unzipping experi- ments in which we derived the average basepair free energy, entropy, and enthalpy of formation of the DNA duplex in a range of temperatures between 5 C and 50 C. We also used the instrument to characterize the temperature-dependent elasticity of single-stranded DNA (ssDNA), where we find a significant condensation plateau at low force and low temperature. Oddly, the persistence length of ssDNA measured at high force seems to increase with temperature, contrary to simple entropic models.The authors thank J. Camunas and S. Frutos for contributing the molecules used in the experiments, and J.M. Huguet for helpful discussion. F.R. is supported by grant Institucio Catalana de Recerca i Estudis Avancats Academia 2013 and J.R. A.-G. by an Explora grant from MINECO (MAT2013-49455-EXP). The research that led to the results presented here was funded by the European Union Seventh Framework Programme (FP7/2007-2013) under grant 308850 INFERNOS and European Research Council grant MagReps (No. 267862).De Lorenzo, S.; Ribezzi-Crivellari, M.; Arias-Gonzalez, JR.; Smith, S.; Ritort, F. (2015). A Temperature-Jump Optical Trap for Single-Molecule Manipulation. Biophysical Journal. 108(12):2854-2864. https://doi.org/10.1016/j.bpj.2015.05.017S285428641081

Simple Detection of Unstained Live Senescent Cells with Imaging Flow Cytometry

Cellular senescence is a hallmark of aging and a promising target for therapeutic approaches. The identification of senescent cells requires multiple biomarkers and complex experimental procedures, resulting in increased variability and reduced sensitivity. Here, we propose a simple and broadly applicable imaging flow cytometry (IFC) method. This method is based on measuring autofluorescence and morphological parameters and on applying recent artificial intelligence (AI) and machine learning (ML) tools. We show that the results of this method are superior to those obtained measuring the classical senescence marker, senescence-associated beta-galactosidase (SA-β-Gal). We provide evidence that this method has the potential for diagnostic or prognostic applications as it was able to detect senescence in cardiac pericytes isolated from the hearts of patients affected by end-stage heart failure. We additionally demonstrate that it can be used to quantify senescence “in vivo” and can be used to evaluate the effects of senolytic compounds. We conclude that this method can be used as a simple and fast senescence assay independently of the origin of the cells and the procedure to induce senescence

Systematic versus on-demand early palliative care: results from a multicentre, randomised clinical trial

Background Early palliative care (EPC) in oncology has been shown to have a positive impact on clinical outcome, quality-of-care outcomes, and costs. However, the optimal way for activating EPC has yet to be defined. Methods This prospective, multicentre, randomised study was conducted on 207 outpatients with metastatic or locally advanced inoperable pancreatic cancer. Patients were randomised to receive ‘standard cancer care plus on-demand EPC’ (n = 100) or ‘standard cancer care plus systematic EPC’ (n = 107). Primary outcome was change in quality of life (QoL) evaluated through the Functional Assessment of Cancer Therapy – Hepatobiliary questionnaire between baseline (T0) and after 12 weeks (T1), in particular the integration of physical, functional, and Hepatic Cancer Subscale (HCS) combined in the Trial Outcome Index (TOI). Patient mood, survival, relatives' satisfaction with care, and indicators of aggressiveness of care were also evaluated. Findings The mean changes in TOI score and HCS score between T0 and T1 were −4.47 and −0.63, with a difference between groups of 3.83 (95% confidence interval [CI] 0.10–7.57) (p = 0.041), and −2.23 and 0.28 (difference between groups of 2.51, 95% CI 0.40–4.61, p = 0.013), in favour of interventional group. QoL scores at T1 of TOI scale and HCS were 84.4 versus 78.1 (p = 0.022) and 52.0 versus 48.2 (p = 0.008), respectively, for interventional and standard arm. Until February 2016, 143 (76.9%) of the 186 evaluable patients had died. There was no difference in overall survival between treatment arms. Interpretations Systematic EPC in advanced pancreatic cancer patients significantly improved QoL with respect to on-demand EPC

"El coloquio de los perros": magic, witchcraft and deceit Critical and literary analysis

reservedAnalisi critica e letteraria della novella di Miguel de Cervantes: “El coloquio de los perros”, attraverso la ricerca e lo studio degli espedienti letterari utilizzati dall’autore che strutturano il racconto sotto forma di un lungo dialogo tra due cani la cui narrazione si sviluppa attraverso la lettura di un manoscritto, opera di un malato di Sifilide che pretende aver riportato ciò che ha udito o forse sognato, durante la sua permanenza presso l’ospedale della Resurreción di Valladolid. L’investigazione si concentra sul tema dell’inganno come forma di vivere e sopravvivere nella società spagnola del ‘600, attraverso la narrazione della vita di Berganza ed il suo servizio come cane, presso diversi padroni. In particolare si mette in relazione il ruolo simbolico del cane, animale fedele e leale ma anche traghettatore di anime, psicopompo, in analogia alla malvagità umana che Berganza ritrova costantemente all'interno di una società alla quale non vuole uniformarsi, giungendo all'unica soluzione di allontanarsi ed isolarsi completamente dalla essa. Infine, si riflette sulla profezia, nucleo centrale della novella, sul suo significato e sul suo possibile compimento attraverso l'analisi delle parole pronunciate da una strega che non solo mette in dubbio l'origine dei due cani ma cambia il significato dell'intera novella, rendendo quella che potrebbe sembrare una semplice storia di animali parlanti, una novella dal significato molto più profondo riguardante l'esistenza umana.Critical and literary analysis of the short story written by Miguel de Cervantes: "El coloquio de los perros", through the research and study of the literary devices used by the author which structure the story in the form of a long dialogue between two dogs whose narration develops through the reading of a manuscript: the work of a syphilis patient who claims to have reported what he heard or perhaps dreamed, during his stay at the Resurreción hospital in Valladolid. The investigation focuses on the theme of deception as a way of living or surviving in 17th century Spanish society, through the narration of Berganza's life and his service as a dog with various masters. In particular, the symbolic role of the dog is related, a faithful and loyal animal but also a ferryman of souls, a psychopomp, in analogy to the human wickedness that Berganza constantly finds within a society to which he does not want to conform, finding the only solution to get away and completely isolate themselves from it. Finally, we reflect on the prophecy, the central core of the novel, on its meaning and on its possible fulfilment through the analysis of the words spoken by a witch who not only questions the origin of the two dogs but changes the meaning of the entire novel, making what might seem like a simple story of talking animals, a novella with much deeper meaning concerning human existence