Controlling liquid-liquid phase behaviour with an active fluid.

Demixing binary liquids is a ubiquitous transition explained using a well-established thermodynamic formalism that requires the equality of intensive thermodynamics parameters across phase boundaries. Demixing transitions also occur when binary fluid mixtures are driven away from equilibrium, but predicting and designing such out-of-equilibrium transitions remains a challenge. Here we study the liquid-liquid phase separation of attractive DNA nanostars driven away from equilibrium using a microtubule-based active fluid. We find that activity lowers the critical temperature and narrows the range of coexistence concentrations, but only in the presence of mechanical bonds between the liquid droplets and reconfiguring active fluid. Similar behaviours are observed in numerical simulations, suggesting that the activity suppression of the critical point is a generic feature of active liquid-liquid phase separation. Our work describes a versatile platform for building soft active materials with feedback control and providing an insight into self-organization in cell biology.

Extending the Scope of the C-Functionalization of Cyclam via Copper(I)-Catalyzed Alkyne-Azide Cycloaddition to Bifunctional Chelators of Interest.

Cyclam, known for its potent chelation properties, is explored for diverse applications through selective N-functionalization, offering versatile ligands for catalysis, medical research, and materials science. The challenges arising from N-alkylation, which could decrease the coordination properties, are addressed by introducing a robust C-functionalization method. The facile two-step synthesis proposed here involves the click chemistry-based C-functionalization of a hydroxyethyl cyclam derivative using Cu(I)-catalyzed alkyne-azide cycloaddition (CuAAC). Boc-protecting groups prevent undesired copper coordination, resulting in compounds with a wide range of functionalities. The optimized synthesis conditions enable C-functional cyclams to be obtained easily and advantageously, with high application potential in the previously cited fields. The methodology has been extended to trehalose-based Siamese twin amphiphiles, enabling efficient gene delivery applications.

Vorticity phase separation and defect lattices in the isotropic phase of active liquid crystals.

We use numerical simulations and linear stability analysis to study the dynamics of an active liquid crystal film on a substrate in the regime where the passive system would be isotropic. Extensile activity builds up local orientational order and destabilizes the quiescent isotropic state above a critical activity, eventually resulting in spatiotemporal chaotic dynamics akin to the one observed ubiquitously in the nematic state. Here we show that tuning substrate friction yields a variety of emergent structures at intermediate activity, including lattices of flow vortices with associated regular arrangements of topological defects and a new state where flow vortices trap pairs of +1/2 defect that chase each other's tail. These chiral units spontaneously pick the sense of rotation and organize in a hexagonal lattice, surrounded by a diffuse flow of opposite rotation to maintain zero net vorticity. The length scale of these emergent structures is set by the screening length of the flow, controlled by the shear viscosity η and the substrate friction Γ, and can be captured by simple mode selection of the vortical flows. We demonstrate that the emergence of coherent structures can be interpreted as a phase separation of vorticity, where friction plays a role akin to that of birth/death processes in breaking conservation of the phase separating species and selecting a characteristic scale for the patterns. Our work shows that friction provides an experimentally accessible tuning parameter for designing controlled active flows.

Activity-Suppressed Phase Separation.

We use a continuum model to examine the effect of activity on a phase-separating mixture of an extensile active nematic and a passive fluid. We highlight the distinct role of (i) previously considered interfacial active stresses and (ii) bulk active stresses that couple to liquid crystalline degrees of freedom. Interfacial active stresses can arrest phase separation, as previously demonstrated. Bulk extensile active stresses can additionally strongly suppress phase separation by sustained self-stirring of the fluid, substantially reducing the size of the coexistence region in the temperature-concentration plane relative to that of the passive system. The phase-separated state is a dynamical emulsion of continuously splitting and merging droplets, as suggested by recent experiments. Using scaling analysis and simulations, we identify various regimes for the dependence of droplet size on activity. These results can provide a criterion for identifying the mechanisms responsible for arresting phase separation in experiments.

Characterization of the perioperative changes of exosomal immune-related cytokines induced by prostatectomy in early-stage prostate cancer patients.

BACKGROUND: Myeloid-derived suppressor cells (MDSCs) are relevant in prostate cancer microenvironment collaborating in tumor development. The main tumor marker used in this disease, prostate-specific antigen (PSA), does not provide information related to this tumor microenvironment. Cancer cells secrete exosomes carrying bioactive molecules contributing to MDSCs recruitment and induction. The aim of this study was to characterize the perioperative changes of exosomal cytokines relevant in MDSCs recruitment induced by prostatectomy in prostate cancer patients. METHODS: Blood was drawn from 26 early-stage prostate cancer patients before and after radical prostatectomy and from 16 healthy volunteers. Serum exosomes were separated by precipitation. Cytokines related with MDSC cell recruitment and activation CCL2, CXCL2, CXCL5, CXCL8, CXCL12, MIF, S100A9 and TGF-ß were measured in serum and serum-derived exosomes using immunometric assays. RESULTS: All cytokines were detected both in serum and exosomes, except for CXCL12, which was detected only in serum. Exosomes were enriched specially in MIF, TGF-ß and CXCL2. Presurgical MIF levels in exosomes correlated negatively with serum PSA. Also, presurgical TGF-ß decreased both in serum and exosomes as Gleason score rises. Patients presurgical exosomes had increased CCL2, CXCL5 and TGF-ß levels than exosomes from healthy controls. These differences were not observed when cytokines were analyzed in serum, except for TGF-ß. Cytokine levels of CCL2, CXCL5 decreased in patients' postsurgical exosomes, while TGF-ß further increased. On the contrary, S100A9 levels were lower in patients presurgical exosomes but increased after radical prostatectomy. CONCLUSIONS: Blood exosomal content in cytokines constitute an attractive source to evaluate MDSCs immunomodulators providing additional information related to tumor microenvironment in prostate cancer.

Stealth Entropy Production in Active Field Theories near Ising Critical Points.

We address the steady-state entropy production rate (EPR) of active scalar ϕ^{4} theories, which lack time-reversal symmetry, close to a phase-separation critical point. We consider both nonconserved (model A) and conserved (model B) dynamics at Gaussian level, and also address the former at leading order in ε=4-d. In each case, activity is irrelevant in the RG sense: the active model lies in the same (dynamic Ising) universality class as its time-reversible counterpart. Hence one might expect that activity brings no new critical behavior. Here we show instead that, on approach to criticality in these models, the singular part of the EPR per (diverging) spacetime correlation volume either remains finite or itself diverges. A nontrivial critical scaling for entropy production thus ranks among universal dynamic Ising-class properties.

A Robust Localization System for Inspection Robots in Sewer Networks.

Sewers represent a very important infrastructure of cities whose state should be monitored periodically. However, the length of such infrastructure prevents sensor networks from being applicable. In this paper, we present a mobile platform (SIAR) designed to inspect the sewer network. It is capable of sensing gas concentrations and detecting failures in the network such as cracks and holes in the floor and walls or zones were the water is not flowing. These alarms should be precisely geo-localized to allow the operators performing the required correcting measures. To this end, this paper presents a robust localization system for global pose estimation on sewers. It makes use of prior information of the sewer network, including its topology, the different cross sections traversed and the position of some elements such as manholes. The system is based on a Monte Carlo Localization system that fuses wheel and RGB-D odometry for the prediction stage. The update step takes into account the sewer network topology for discarding wrong hypotheses. Additionally, the localization is further refined with novel updating steps proposed in this paper which are activated whenever a discrete element in the sewer network is detected or the relative orientation of the robot over the sewer gallery could be estimated. Each part of the system has been validated with real data obtained from the sewers of Barcelona. The whole system is able to obtain median localization errors in the order of one meter in all cases. Finally, the paper also includes comparisons with state-of-the-art Simultaneous Localization and Mapping (SLAM) systems that demonstrate the convenience of the approach.

Strong Coupling in Conserved Surface Roughening: A New Universality Class?

The Kardar-Parisi-Zhang (KPZ) equation defines the main universality class for nonlinear growth and roughening of surfaces. But under certain conditions, a conserved KPZ equation (CKPZ) is thought to set the universality class instead. This has non-mean-field behavior only in spatial dimension d<2. We point out here that CKPZ is incomplete: It omits a symmetry-allowed nonlinear gradient term of the same order as the one retained. Adding this term, we find a parameter regime where the one-loop renormalization group flow diverges. This suggests a phase transition to a new growth phase, possibly ruled by a strong-coupling fixed point and thus described by a new universality class, for any d>1. In this phase, numerical integration of the model in d=2 gives clear evidence of non-mean-field behavior.

Prevalence analysis of urinary incontinence after radical prostatectomy and influential preoperative factors in a single institution.

AIMS: To assess prevalence of urinary incontinence (UI) after radical prostatectomy (RP) and to analyze which preoperative characteristics of the patients have influence on UI. METHODS: Between 2002 and 2012, 746 consecutive patients underwent RP for clinically localized prostate cancer. We defined UI according to International Continence Society (ICS) definition: "the complaint of any involuntary leakage of urine" after 12 months of recovery, international consultation on incontinence questionnaire (ICIQ-SF) and pads/day was collected too. Clinical features and magnetic resonance imaging measurements were assessed. A multivariable logistic regression model predicting incontinence were built-in after adjust by cofounding factors and bootstrapping. RESULTS: About 172 (23%) of the patients were classified as incontinent according to the ICS definition. The mean value of the ICIQ-SF was 10.87 (±4). 17.8% of patients use at least one pad/day, 11.9% use more than one pad/day. The preoperative factors independently influential in UI are: age [OR: 1.055; CI 95% (1.006-1.107), p = .028], urethral wall thickness [OR: 5.03; CI 95% (1.11-22.8), p = .036], history of transurethral resection of the prostate [OR: 6.13; CI 95% (1.86-20.18), p = .003] and membranous urethral length [OR: 0.173; CI 95% (0.046-0.64), p = .009]. The predictive accuracy of the model is 78.7% and the area under the curve (AUC) value 71.7%. CONCLUSIONS: Urinary incontinence after radical prostatectomy has different prevalence depending on the definition. Age, prior transurethral resection of the prostate (TURP), membranous urethral length (MUL) and urethral wall thickness (UWT) were risk factors.

Validation of the Jefferson Scale of Physician Empathy in Spanish medical students who participated in an Early Clerkship Immersion programme.

BACKGROUND: The Jefferson Scale of Physician Empathy is the most widely used instrument to measure empathy in the doctor-patient relationship. This work pursued cultural adaptation and validation of the original scale, in its health professions version (JSE-HP), for medical students who participate in an Early Clerkship Immersion Programme of a Spanish university. METHODS: The questionnaire was replied by 506 1st, 2nd, 3rd and 5th year medical students from Universidad Francisco de Vitoria, Madrid, in 2014 and 2016. Internal consistency was analysed by means of Cronbach's alpha, and reliability by means of test-retest using the intraclass correlation coefficient and the Bland-Altman method. The construct validity was checked by means of confirmatory factor analysis and association with other empathy-related variables. Criterion validity was compared using Davis' Interpersonal Reactivity Index. RESULTS: Cronbach's alpha was 0.82 (range 0.80-0.85). Item-total score correlations were positive and significant (median 0.45, p <  0.01). The test-retest intraclass correlation coefficient was 0.68 (0.42-0.82). The factor analysis confirmed the three original factors: "perspective taking", "compassionate care" and "standing in the patient's shoes". Women and students who preferred specialities focused on persons obtained the best scores. The JSE-HP scores were positively correlated with Interpersonal Reactivity Index, personality traits were associated with empathy, clinical interview skills and Objective Structured Clinical Examinations. CONCLUSION: The results support the validity and reliability of JSE-HP applied to Spanish medical students.

The Impact of Heteromultivalency in Lectin Recognition and Glycosidase Inhibition: An Integrated Mechanistic Study.

The vision of multivalency as a strategy limited to achieve affinity enhancements between a protein receptor and its putative sugar ligand (glycotope) has proven too simplistic. On the one hand, binding of a glycotope in a dense glycocalix-like construct to a lectin partner has been shown to be sensitive to the presence of a third sugar entity (heterocluster effect). On the other hand, several carbohydrate processing enzymes (glycosidases and glycosyltransferases) have been found to be also responsive to multivalent presentations of binding partners (multivalent enzyme inhibition), a phenomenon first discovered for iminosugar-type inhibitory species (inhitopes) and recently demonstrated for multivalent carbohydrate constructs. By assessing a series of homo- and heteroclusters combining α-d-glucopyranosyl-related glycotopes and inhitopes, it was shown that multivalency and heteromultivalency govern both kinds of events, allowing for activation, deactivation or enhancement of specific recognition phenomena towards a spectrum of lectin and glycosidase partners in a multimodal manner. This unified scenario originates from the ability of (hetero)multivalent architectures to trigger glycosidase binding modes that are reminiscent of those harnessed by lectins, which should be considered when profiling the biological activity of multivalent architectures.

A Latin American, Portuguese and Spanish consensus on a core communication curriculum for undergraduate medical education.

BACKGROUND: To present learning outcomes in clinical communication for a Core Curriculum for medical undergraduate students in Latin America, Portugal and Spain (LAPS-CCC) and to establish an expert network to support a transnational implementation. METHODS: Through an iterative process, an international group of 15 experts developed an initial set of learning outcomes following a review and discussion of relevant international and local literature. A two-round Delphi survey involving 46 experts from 8 countries was performed. Quantative and qualitative analisis permited the definition of the final consensus. RESULTS: The initial proposal included 157 learning outcomes. The Delphi process generated 734 comments and involved the modification, deletion and addition of some outcomes. At the end of the process, a consensus was reached on 136 learning outcomes grouped under 6 competency domains with a high overall acceptance (95.1 %). CONCLUSIONS: The learning outcomes of this proposal provide a guide to introduce, support and develop communication curriculae for undergraduate medical studies in the countries involved or in other Spanish- or Portuguese-speaking countries.

Radical prostatectomy. Detailed surgical margins. Prognostic value of multifocal involvement in pT2 (+).

OBJECTIVES: We intend to analyze the prognostic value of positive surgical margins depending on their number and location in pT2 patients. METHODS: We analyze 448 (34.3%) patients with positive surgical margins from a series of 1,310 T1-T2 patients treated with radical prostatectomy between 1989-2012. Of them 164 are pT2 (+). 119 (72.6% ) have unifocal affectation (41 (34.5%) unifocal in right lobe; 35 (29.4%) unifocal in left lobe, 40 (33.6%) unifocal in apex, 3 (2.5% ) unifocal proximal) and 45 (27.4%) multifocal involvement. RESULTS: Unifocal and multifocal pT2(+)patients have not evidenced significant differences in any of the clinicopathologic variables compared. However the BPFS at 5 and 10 years is significantly worse in the multifocal group, (p<0.000) In the BPFS multivariate study of 164 pT2(+ )influential variables are: multifocal involvement (HR: 3.4; 95%IC 1.7-6.9 p<0.000) and PSA (HR: 1.03; 95%IC 1.02-1.05 p<0.000), being PSA >15 ng/ml )HR: 3.7; 95%IC 2.1-6.6 p<0.000 ( the best cut-off point. Risk groups: Using the independent influence variables, the best model (using Cox models ) includes two risk groups: Group 1 (0 variables): They are pT2(+) with unifocal affectation and PSA<15 ng/ml, (63%). Their BPFS are 81±4% and 77±4% (5 and 10 years). Grupo 2 (1-2 variables): They are pT2 (+) with multifocal involvement, PSA> 15 ng/ml or both of them, (37%). Their BPFS are 46±6% and 26±7% (5 and 10 years). The BPFS differs significantly between the two groups (p<0.000). The Group 1 BPFS is similar to the pT2 (-) patients, (p:0.242). The Group 2 BPFS is similar to the pT3(+) patients, (p:0.637). The model explained significantly better the BPFS than any of the individual variables analyzed. CONCLUSIONS: In pT2(+) patients the prognosis is significantly worse in multifocal involvement. In addition two groups of patients can be clearly distinguished from the BPFS point view according to their influential variables. The data suggest that since the prognostic point view the second group is understaged while the first is overstaged.

Trehalose-polyamine/DNA nanocomplexes: impact of vector architecture on cell and organ transfection selectivity.

A novel family of precision-engineered gene vectors with well-defined structures built on trehalose and trehalose-based macrocycles (cyclotrehalans) comprising linear or cyclic polyamine heads have been synthesized through procedures that exploit click chemistry reactions. The strategy was conceived to enable systematic structural variations and, at the same time, ensuring that enantiomerically pure vectors are obtained. Notably, changes in the molecular architecture translated into topological differences at the nanoscale upon co-assembly with plasmid DNA, especially regarding the presence of regions with short- or long-range internal order as observed by TEM. In vitro and in vivo experiments further evidenced a significant impact on cell and organ transfection selectivity. Altogether, the results highlight the potential of trehalose-polyamine/pDNA nanocomplex monoformulations to achieve targeting transfection without the need for any additional cell- or organ-sorting component.

Short-term cultured tumor fragments to study immunotherapy combinations based on CD137 (4-1BB) agonism.

Biomarkers for cancer immunotherapy are an unmet medical need. The group of Daniela Thommen at the NKI recently reported on novel methodologies based on short-term cultures of patient-derived tumor fragments whose cytokine concentrations in the supernatants and activation markers on infiltrating T cells were associated with clinical response to PD-1 blockade. We set up a similar culture technology with tumor-derived fragments using mouse tumors transplanted into syngeneic immunocompetent mice to test an agonist anti-CD137 mAb and its combinations with anti-PD-1 and/or anti-TGF-ß. Increases in IFNγ concentrations in the tissue culture supernatants were detected upon in-culture activation with the anti-CD137 and anti-PD-1 mAb combinations or concanavalin A as a positive control. No other cytokine from a wide array was informative of stimulation with these mAbs. Interestingly, increases in Ki67 and other activation markers were substantiated in lymphocytes from cell suspensions gathered at the end of 72 h cultures. In mice bearing bilateral tumors in which one was excised prior to in vivo anti-CD137 + anti-PD-1 treatment to perform the fragment culture evaluation, no association was found between IFNγ production from the fragments and the in vivo therapeutic outcome in the non-resected contralateral tumors. The experimental system permitted freezing and thawing of the fragments with similar functional outcomes. Using a series of patient-derived tumor fragments from excised solid malignancies, we showed IFNγ production in a fraction of the studied cases, that was conserved in frozen/thawed fragments. The small tumor fragment culture technique seems suitable to preclinically explore immunotherapy combinations.

Asthma control and concordance of opinions between patients and pulmonologists.

UNLABELLED: Patient-physician opinion concordance could play a key role in asthma control. There have been no studies evaluating this association in large samples of patients. OBJECTIVES: To determine opinion concordance between asthma patients and their pulmonologists on the impact of the disease and to correlate concordance to asthma control. METHODS: This was a cross-sectional multicentre study including 1160 patients and 300 pulmonologists. Patient-physician concordance rates were assessed by two semi-structured qualitative questionnaires: (1) impact of the disease and (2) treatment satisfaction. Subsequently, participating pulmonologists determined the concordance between their perceptions and their patient's. Sociodemographic and clinical data were recorded for all patients. RESULTS: In 53.6% of cases, asthma was controlled. The rate of patient-pulmonologist concordance on disease impact on patient daily life was 57%, with physicians underestimating the impact (compared to patients) in 26% of cases. Concordance on satisfaction with treatment was 56%, with physicians underestimating satisfaction in 26% of cases. Patient-physician discordance rates were significantly lower among patients with controlled asthma (29 and 32.1%) than those with poor control (73.7 and 73.1%). CONCLUSIONS: Patient-pulmonologist concordance on perceptions of disease impact is low, particularly in uncontrolled asthma. This poor concordance should be addressed in education programmes, particularly for patients with uncontrolled symptoms.

General infection prevention, mitigation, and control procedures implemented in the university education during the COVID-19 pandemic to achieve classroom attendance: a successful community case study.

Introduction: The COVID-19 pandemic entailed confinement and elimination of face-to-face university classes in Spain. The Francisco de Vitoria University in Madrid (UFV by its Spanish acronym) implemented risk management systems to enable on-campus university activity to avoid a negative impact on students, teachers, and faculties. Methods: A tracking/registry system was implemented to collect data, identify COVID-19-related cases, implement containment measures, and do follow-up in the UFV community (administration/services personnel [ASP], teaching/research personnel [TRP], and students), from September 2020 to April 2022. In addition, a prevention plan was implemented on campus to avoid COVID-19 spreading. Satisfaction with these measures was assessed through an online questionnaire. Results: A total of 7,165 suspected COVID-19 cases (84.7% students, 7.7% ASP, 6.5% TRP) were tracked (62.5% female cases, mean age (±SD) 24.8 years (±9.2 years)), and 45% of them confirmed (82% symptomatic/16% asymptomatic), being the student group that with the highest percentage (38.3% total tracked cases). The source of infection was identified in 50.6% of the confirmed cases (90.2% located off-campus). Nineteen COVID-19 outbreaks were registered (inside-10/outside-9). COVID-19 incidence rates were similar or lower than those reported in the Community of Madrid, except in the last wave, corresponding to Omicron variant. The degree of satisfaction (scale 1-6) with the implemented measures was high (scores 4.48-5.44). Conclusion: During the COVID-19 pandemic, UFV control measures, periodic monitoring, and the effectiveness of the tracking system have contributed to maintaining classroom teaching, guaranteeing health and safety. UFV has adapted to a new reality as an example of good practice for future pandemics or emergency situations.

Disruption of the Arabidopsis thaliana inward-rectifier K+ channel AKT1 improves plant responses to water stress.

The Arabidopsis thaliana inward-rectifier K(+) channel AKT1 plays an important role in root K(+) uptake. Recent results show that the calcineurin B-like (CBL)-interacting protein kinase (CIPK) 23-CBL1/9 complex activates AKT1 in the root to enhance K(+) uptake. In addition, this CIPK-CBL complex has been demonstrated to regulate stomatal movements and plant transpiration. However, a role for AKT1 in plant transpiration has not yet been demonstrated. Here we show that disruption of AKT1 conferred an enhanced response to water stress in plants. Experiments performed in hydroponics showed that, when water potential was diminished by adding polyethylene glycol, akt1 adult plants lost less water than wild-type (WT) plants. Under long-term water stress in soil, adult akt1 plants displayed lower transpiration and less water consumption than WT plants. Finally, akt1 stomata closed more efficiently in response to ABA. Such results were also observed in cipk23 plants. The similar responses shown by cipk23 and akt1 plants to water stress denote that the regulation of AKT1 by CIPK23 may also take place in stomata and has a negative impact on plant performance under water stress conditions.

A Ca(2+)-sensitive system mediates low-affinity K(+) uptake in the absence of AKT1 in Arabidopsis plants.

K(+) acquisition by Arabidopsis roots is mainly mediated by the high-affinity K(+) transporter AtHAK5 and the inward-rectifier K(+) channel AtAKT1. This model is probably universal to plants. Mutant plants lacking these two systems (athak5,atakt1) take up K(+) and grow when the external K(+) concentration is above a certain level, indicating that an additional transport system may compensate for the absence of AtHAK5 and AtAKT1. Here we describe that this alternative system is essential for providing sufficient K(+) to sustain growth of athak5,atakt1 plants. This system is especially sensitive to Ca(2+), Mg(2+), Ba(2+) and La(3+), it transports Cs(+) and its activity is reduced by cyclic nucleotides. These results suggest that a Ca(2+)-permeable voltage-independent non-selective cation channel, probably belonging to the cyclic nucleotide gated channel (CNGC) family, may provide the pathway for K(+) uptake in athak5,atakt1 plants. The genes encoding the two members of the CNGC family that have been described as mediating root K(+) uptake, AtCNGC3 and AtCNGC10, are not up-regulated in athak5,atakt1 plants, excluding overexpression of these genes as a compensatory mechanism. On the other hand, an increased driving force for K(+) in athak5,atakt1 plants due to a hyperpolarization of the membrane potential of its root cells is also discarded. The identification of this unknown system may provide tools to improve plant K(+) nutrition in conditions where AtAKT1 functionality is reduced, such as under salinity. In addition, this system may constitute an important pathway for accumulation of toxic cations such as Cs(+) or radiocesium ((137)Cs(+)), and could play a role in phytoremediation.