A Biobank of Breast Cancer Explants with Preserved Intra-tumor Heterogeneity to Screen Anticancer Compounds.

The inter- and intra-tumor heterogeneity of breast cancer needs to be adequately captured in pre-clinical models. We have created a large collection of breast cancer patient-derived tumor xenografts (PDTXs), in which the morphological and molecular characteristics of the originating tumor are preserved through passaging in the mouse. An integrated platform combining in vivo maintenance of these PDTXs along with short-term cultures of PDTX-derived tumor cells (PDTCs) was optimized. Remarkably, the intra-tumor genomic clonal architecture present in the originating breast cancers was mostly preserved upon serial passaging in xenografts and in short-term cultured PDTCs. We assessed drug responses in PDTCs on a high-throughput platform and validated several ex vivo responses in vivo. The biobank represents a powerful resource for pre-clinical breast cancer pharmacogenomic studies (http://caldaslab.cruk.cam.ac.uk/bcape), including identification of biomarkers of response or resistance.

Telemedicine in Orthopedic Surgery: A Systematic Review of Current Evidence.

Introduction: The coronavirus disease 2019 (COVID-19) pandemic has triggered transformative changes in how clinicians interact with patients. There has been a shift toward virtual consultations. The evidence to support this change in practice is unclear. The aim of this study was to systematically review the evidence base for virtual consultations for orthopedics. Materials and Methods: Two independent reviewers performed a literature search based on Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, utilizing the MEDLINE, EMBASE, and Scopus databases. Only studies reporting outcomes following the use of telemedicine for diagnosis, consultation, rehabilitation, and follow-up were included. Outcomes analyzed were: (1) patient and clinician satisfaction, (2) clinical outcome measures, and (3) cost analysis of traditional versus teleconsultation. Results: A total of 41 studies were included. Fifteen studies compared clinical outcomes of telemedicine against a matched traditional cohort. Of these 15 studies, 2 demonstrated noninferiority, 9 showed no statistically significant difference, and 4 found telemedicine to be superior. Eleven studies recorded patient reported outcomes, which demonstrated high patient satisfaction. Nine studies reported decreased costs when telemedicine was compared to traditional care. The remaining six studies had varied aims and methodologies that didn't fit well with any of these subheadings. Discussion: While the available evidence is limited, the studies assessed here show that telemedicine can deliver high quality health care with good clinical outcomes and high patient satisfaction in a cost-effective manner. Our team thinks what this has highlighted is that communication technology is advancing rapidly and that we as a community of surgeons need to be able to adapt rapidly and adopt innovative technology to continue to improve patient experience and outcomes.

Use of confocal Raman microscopy to characterise ethyl cyanoacrylate adhesive depth curing.

In situ spatial temporal measurement of monomer conversion during adhesive bondline curing remains a challenging area. The aim of this work was to demonstrate the effectiveness of using confocal Raman microscopy in a specially configured experimental set-up, as a versatile tool for measuring monomer concentration changes as a function of both time and adhesive bond depth during ethyl cyanoacrylate polymerisation. This also allowed monitoring of the extent of polymerisation at the adhesive substrate interface independently of the bulk bondline polymerisation region. Key kinetic parameters such as inhibition time tlag, rate of reaction Rmax and extent of reaction [αt]max were obtained by fitting the experimental data to sigmoidal growth curves using simple piecewise regression models. A systematic characterisation of a polymerisation reaction was conducted using different sample substrate types (copper alloy (red brass), aluminium, aluminium alloy, stainless steel and borosilicate glass) and at various reaction temperatures. Reaction rates were found to decrease further away from the substrate interface in the bulk volume region. The fastest kinetics occurred in the vicinity of nucleophilic hydroxyl rich surfaces such as at the copper alloy (red brass). In addition to substrate surface chemistry, surface roughness was also a factor, with the highest reaction rates occurring with a grit blasted (roughened) aluminium alloy (2024 T3) surface. An approximately linear dependence of the ln Rmaxvs. 1/T (Arrhenius) plot was recorded within the temperature range of 291-328 K. A better fit was obtained however through the use of two separate linear slopes, possibly indicative of a change of polymerisation reaction mechanism taking place at elevated temperatures with two distinct activation energies. Further work conducted using a larger number of temperatures would be useful to verify this finding. This work confirmed that differences in the rates of interfacial and bulk polymerisation processes could be readily measured in situ using confocal Raman microscopy which is a powerful technique for investigating such surface-confined and bulk polymerisation reactions.

Formation of p-n junctions in nanoparticle cerium oxide electrolytic cells displaying memristive switching behaviour.

A macro-scale metal-semiconductor-metal device comprising CeO2 nanoparticles cast from a suspension of cerium dioxide formed by a novel synthetic method was fabricated. Thin CeO2 films of 40 nm thickness placed between panels of aluminium and/or copper displayed memristive-like resistive switching behaviour upon the application of potential sweeps ranging between -0.6 V and 0.6 V. A mechanism is proposed based on the notion that an electrolytic cell operates under such conditions with the initial formation of p and n-type regions within the central semiconductive thin film. Evidence is presented for the existence of numerous point defects in these nanosized CeO2 films, which are also likely to play a role in the device's operation acting as internal dopants. Steady currents were observed upon the imposition of constant potentials, most notably at higher potential values (both anodic and cathodic). It is suggested that electrons and holes act as charge carriers in these devices rather than ionic species as proposed in some other mechanisms.

Modified Surface Relief Layer Created by Holographic Lithography: Application to Selective Sodium and Potassium Sensing.

Point-of-care diagnostics will rely upon the development of low-cost, noncomplex, and easily integrated systems in order to examine biological samples such as blood and urine obtained from the patient. The development of metal ion sensors is a subject of significant relevance for physiological samples. The level of different blood electrolytes, mainly H⁺, Na⁺, K⁺ and Cl- is considerably used to monitor irregular physiologies. The particular challenge in biosensing, and in fact for any other sensor, is signal differentiation between non-specifically bound material and the specific detecting of the target molecule/ion. The biosensors described in this paper are fabricated by a holographic recording of surface relief structures in a photopolymer material. The surface structures are modified by coating with either dibenzo-18-crown-6 (DC) or tetraethyl 4-tert-butylcalix[4]arene (TBC), which are embedded in a polymer matrix. Interrogation of these structures by light allows indirect measurement of the concentration of the analyte. The influence of polymer matrices with different porosities, plasticised polyvinyl chloride (PVC) and a sol-gel matrix, on the performance of the sensors for detection of K⁺ and Na⁺ is examined. Here we demonstrate a proof of concept that by using a matrix with higher porosity one can increase the sensitivity of the sensor. The results showed that the DC sensing layer provides a selective response to K⁺ over Na⁺ and the TBC modified grating is more responsive to Na⁺ over K⁺. The sensor responds to K⁺ and Na⁺ within the physiological concentration ranges.

Self-processing photopolymer materials for versatile design and fabrication of holographic sensors and interactive holograms.

The aim of this paper is to discuss the benefits as well as the limitations of utilizing photopolymer materials in the design of holograms that are responsive to changes in their environment, such as changes in the concentration of a specific substance, temperature, and pressure. Three different case studies are presented, including both surface and volume phase holograms, in order to demonstrate the flexibility in the approach of utilizing holographic photopolymers for the design of sensors and interactive optical devices. First, a functionalized surface relief hologram is demonstrated to operate as an optical sensor for the detection of metal ions in water. The sensitivity and selectivity of the sensor are investigated. The second example demonstrates a volume transmission hologram recorded in a temperature-sensitive photopolymer and the memory effects of its exposure to elevated temperature. Finally, a pressure-sensitive reflection hologram that changes color under application of pressure is characterized, and its potential application in document authentication is described.

An unusual pattern of peritubular capillary injury involving apoptosis in a renal transplant patient.

Microvascular injury is an important factor in renal allograft survival. Repeated episodes of endothelial injury from chronic antibody-mediated rejection typically manifest at the ultrastructural level as circumferential multilayering of remodeled glomerular basement membrane material and peritubular capillary basal lamina. In contrast to this typical pattern of microvascular injury, a renal transplantation case is presented in which focally dilated and multilayered segments of peritubular capillary basal lamina bearing lipid droplets were interspersed with ultrastructurally normal unilayered segments of basal lamina devoid of lipid droplets. Glomerular basement membranes were not affected by this process. The peak incidence of lipid droplets within the peritubular capillary walls coincided with a peak in apoptotic activity within the allograft. Lesser amounts of the same lipidic material were identified in the mesangial matrix and an arteriolar wall. Mesangial electron-dense deposits were detected at two weeks posttransplantation and their appearance coincided with elevated immunological activity in the glomeruli, as determined by immunofluorescence microscopy. The unusual ultrastructure and immunological activity observed in this case may reflect a process of impaired apoptotic clearance within the allograft. The six biopsies from a single patient are discussed in the setting of a highly sensitized renal transplant recipient who received prophylactic terminal complement blockade by eculizumab. The findings may be relevant to the study of apoptosis, efferocytosis, microvascular injury, eculizumab, rejection, lupus, and drug-related disease.

Functional exofacially tagged N-type calcium channels elucidate the interaction with auxiliary α2δ-1 subunits.

CaV1 and CaV2 voltage-gated calcium channels are associated with ß and α2δ accessory subunits. However, examination of cell surface-associated CaV2 channels has been hampered by the lack of antibodies to cell surface-accessible epitopes and of functional exofacially tagged CaV2 channels. Here we report the development of fully functional CaV2.2 constructs containing inserted surface-accessible exofacial tags, which allow visualization of only those channels at the plasma membrane, in both a neuronal cell line and neurons. We first examined the effect of the auxiliary subunits. Although α2δ subunits copurify with CaV2 channels, it has recently been suggested that this interaction is easily disrupted and nonquantitative. We have now tested whether α2δ subunits are associated with these channels at the cell surface. We found that, whereas α2δ-1 is readily observed at the plasma membrane when expressed alone, it appears absent when coexpressed with CaV2.2/ß1b, despite our finding that α2δ-1 increases plasma-membrane CaV2.2 expression. However, this was due to occlusion of the antigenic epitope by association with CaV2.2, as revealed by antigen retrieval; thus, our data provide evidence for a tight interaction between α2δ-1 and the α1 subunit at the plasma membrane. We further show that, although CaV2.2 cell-surface expression is reduced by gabapentin in the presence of wild-type α2δ-1 (but not a gabapentin-insensitive α2δ-1 mutant), the interaction between CaV2.2 and α2δ-1 is not disrupted by gabapentin. Altogether, these results demonstrate that CaV2.2 and α2δ-1 are intimately associated at the plasma membrane and allow us to infer a region of interaction.

The upregulation of α2δ-1 subunit modulates activity-dependent Ca2+ signals in sensory neurons.

As auxiliary subunits of voltage-gated Ca(2+) channels, the α2δ proteins modulate membrane trafficking of the channels and their localization to specific presynaptic sites. Following nerve injury, upregulation of the α2δ-1 subunit in sensory dorsal root ganglion neurons contributes to the generation of chronic pain states; however, very little is known about the underlying molecular mechanisms. Here we show that the increased expression of α2δ-1 in rat sensory neurons leads to prolonged Ca(2+) responses evoked by membrane depolarization. This mechanism is coupled to CaV2.2 channel-mediated responses, as it is blocked by a ω-conotoxin GVIA application. Once initiated, the prolonged Ca(2+) transients are not dependent on extracellular Ca(2+) and do not require Ca(2+) release from the endoplasmic reticulum. The selective inhibition of mitochondrial Ca(2+) uptake demonstrates that α2δ-1-mediated prolonged Ca(2+) signals are buffered by mitochondria, preferentially activated by Ca(2+) influx through CaV2.2 channels. Thus, by controlling channel abundance at the plasma membrane, the α2δ-1 subunit has a major impact on the organization of depolarization-induced intracellular Ca(2+) signaling in dorsal root ganglion neurons.

Electron-dense deposit in renal transplant patients on eculizumab may be drug-derived.

Eculizumab is a monoclonal antibody that inhibits the conversion of complement protein C5 to C5a and C5b. Eculizumab has been used to treat some disorders of complement regulation owing to its ability to inhibit terminal complement activation. The efficacy of eculizumab in reducing complement-mediated microvascular injury in renal allografts is currently the subject of trials. Electron-dense deposit was detected in allograft biopsies from three highly sensitized recipients of renal transplants, all of whom had received prophylactic eculizumab therapy. In two cases, the deposit was probably drug-derived whilst in the third case the deposit was probably derived from recurrent disease. The cases demonstrate the potential difficulty in interpreting electron-dense deposit in renal allograft biopsies, particularly in the setting of eculizumab therapy.

Return-to-work challenges following a work-related mild TBI: The injured worker perspective.

PRIMARY OBJECTIVE: To explore how individuals with work-related mild traumatic brain injury (wrMTBI) experience return-to-work (RTW) processes when returning to the workplace where the injury occurred. DESIGN: RTW experiences were explored using in-depth interviews and an inductive analytic approach. Qualitative analysis guided by the research question moved through phases of line-by-line and thematic coding through which categories and the interaction between categories emerged. PARTICIPANTS: Twelve workers diagnosed with a wrMTBI reported on their RTW experiences following wrMTBIs that occurred 3-5 years prior to the time of the interview. MAIN OUTCOMES AND RESULTS: Participants perceived employer and workers' compensation factors as profoundly influencing their RTW experiences. Participants consistently reported that employers and workers' compensation representatives had an inadequate understanding of wrMTBI sequelae. Six of 12 participants were re-injured following their wrMTBI, with three of these injuries occurring at work. CONCLUSION: Employers, co-workers and workers' compensation representatives should be aware of wrMTBI sequelae so injured workers can receive appropriate supports and both stigmatization and re-injury can be mitigated. Greater attention to the structural and social elements of workplace and compensation environments could inform strategies to break down barriers to successful return-to-work following a wrMTBI.

Medium-Term Clinical Outcomes of the Global Icon Stemless Shoulder System: Results of a 2-Year Follow-Up.

The primary aim of this study was to assess the medium-term outcomes of the Global Icon stemless shoulder replacement in patients who have undergone primary total shoulder arthroplasty (TSA) for glenohumeral joint osteoarthritis. A retrospective review of patients who had undergone a TSA using the Global Icon stemless shoulder system was performed. The Western Ontario Osteoarthritis Shoulder (WOOS) Index and Oxford Shoulder Score (OSS) were evaluated pre-operatively and at 12 to 24 months post-operatively. Radiological outcomes, operation time, and post-operative complications were reported. Primary analysis for the WOOS Index and OSS focused on detecting within-group treatment effects at 24 months using a repeated measures ANOVA. Thirty patients were included in the study. Post-surgery, there was a significant improvement at 24 months on the OSS (ES = 0.932, CI: 41.7 to 47.7, p < 0.001) and the WOOS Index (ES = 0.906, CI: 71.9 to 99.8, p < 0.001). Radiographs revealed that no component loosened, migrated, or subsided. The median operative time was 75.5 (IQR: 12.25, range: 18 to 105) min. No implant-related complications were reported. The Global Icon stemless replacements have excellent clinical outcomes in this cohort at 12- and 24-month follow-up with no implant-related complications.

Common health assets protocol: a mixed-methods, realist evaluation and economic appraisal of how community-led organisations (CLOs) impact on the health and well-being of people living in deprived areas.

INTRODUCTION: This research investigates how community-led organisations' (CLOs') use of assets-based approaches improves health and well-being, and how that might be different in different contexts. Assets-based approaches involve 'doing with' rather than 'doing to' and bring people in communities together to achieve positive change using their own knowledge, skills and experience. Some studies have shown that such approaches can have a positive effect on health and well-being. However, research is limited, and we know little about which approaches lead to which outcomes and how different contexts might affect success. METHODS AND ANALYSIS: Using a realist approach, we will work with 15 CLOs based in disadvantaged communities in England, Scotland and Northern Ireland. A realist synthesis of review papers, and a policy analysis in different contexts, precedes qualitative interviews and workshops with stakeholders, to find out how CLOs' programmes work and identify existing data. We will explore participants' experiences through: a Q methodology study; participatory photography workshops; qualitative interviews and measure outcomes using a longitudinal survey, with 225 CLO participants, to assess impact for people who connect with the CLOs. An economic analysis will estimate costs and benefits to participants, for different contexts and mechanisms. A 'Lived Experience Panel' of people connected with our CLOs as participants or volunteers, will ensure the appropriateness of the research, interpretation and reporting of findings. ETHICS AND DISSEMINATION: This project, research tools and consent processes have been approved by the Glasgow Caledonian University School of Health and Life Sciences Ethics Committee, and affirmed by Ethics Committees at Bournemouth University, Queen's University Belfast and the University of East London. Common Health Assets does not involve any National Health Service sites, staff or patients.Findings will be presented through social media, project website, blogs, policy briefings, journal articles, conferences and visually in short digital stories, and photographic exhibitions.

Non aggregated colloidal silver nanoparticles for surface enhanced resonance Raman spectroscopy.

Silver nanoparticles with a tuneable λ max were produced as colloids by heterogeneous nucleation. The synthesis process is both fast and repeatable, producing stable PVA capped nanoparticles. The colloid's effectiveness in the SERRS system was investigated using Rhodamine 6G, R6G, Crystal Violet, CV, and Malachite Green, MG, as probe molecules. A clear sensing trend was observed, where the Raman signal emitted was significantly enhanced by the addition of silver nanoparticles. A build up of signal intensity is observed until an optimum ratio is achieved, followed by a decline in signal intensity as the concentration of nanoparticles is further increased. The sensing trend appeared to be dependant on the structure of these model molecules with similarly structured compounds exhibiting similar trends. Thus a maximum enhancement with the Ag: analyte molar ratio of ∼ 5.56: 1, was seen for CV and MG whereas R6G had a maximum enhancement at the Ag: analyte molar ratio of ∼ 2.25: 1.

A wide depth distribution of seismic tremors along the northern Cascadia margin.

The Cascadia subduction zone is thought to be capable of generating major earthquakes with moment magnitude as large as M(w) = 9 at an interval of several hundred years. The seismogenic portion of the plate interface is mostly offshore and is currently locked, as inferred from geodetic data. However, episodic surface displacements-in the direction opposite to the long-term deformation motions caused by relative plate convergence across a locked interface-are observed about every 14 months with an unusual tremor-like seismic signature. Here we show that these tremors are distributed over a depth range exceeding 40 km within a limited horizontal band. Many occurred within or close to the strong seismic reflectors above the plate interface where local earthquakes are absent, suggesting that the seismogenic process for tremors is fluid-related. The observed depth range implies that tremors could be associated with the variation of stress field induced by a transient slip along the deeper portion of the Cascadia interface or, alternatively, that episodic slip is more diffuse than originally suggested.

Identifying Cancer Drivers Using DRIVE: A Feature-Based Machine Learning Model for a Pan-Cancer Assessment of Somatic Missense Mutations.

Sporadic cancer develops from the accrual of somatic mutations. Out of all small-scale somatic aberrations in coding regions, 95% are base substitutions, with 90% being missense mutations. While multiple studies focused on the importance of this mutation type, a machine learning method based on the number of protein-protein interactions (PPIs) has not been fully explored. This study aims to develop an improved computational method for driver identification, validation and evaluation (DRIVE), which is compared to other methods for assessing its performance. DRIVE aims at distinguishing between driver and passenger mutations using a feature-based learning approach comprising two levels of biological classification for a pan-cancer assessment of somatic mutations. Gene-level features include the maximum number of protein-protein interactions, the biological process and the type of post-translational modifications (PTMs) while mutation-level features are based on pathogenicity scores. Multiple supervised classification algorithms were trained on Genomics Evidence Neoplasia Information Exchange (GENIE) project data and then tested on an independent dataset from The Cancer Genome Atlas (TCGA) study. Finally, the most powerful classifier using DRIVE was evaluated on a benchmark dataset, which showed a better overall performance compared to other state-of-the-art methodologies, however, considerable care must be taken due to the reduced size of the dataset. DRIVE outlines the outstanding potential that multiple levels of a feature-based learning model will play in the future of oncology-based precision medicine.

RAC1B modulates intestinal tumourigenesis via modulation of WNT and EGFR signalling pathways.

Current therapeutic options for treating colorectal cancer have little clinical efficacy and acquired resistance during treatment is common, even following patient stratification. Understanding the mechanisms that promote therapy resistance may lead to the development of novel therapeutic options that complement existing treatments and improve patient outcome. Here, we identify RAC1B as an important mediator of colorectal tumourigenesis and a potential target for enhancing the efficacy of EGFR inhibitor treatment. We find that high RAC1B expression in human colorectal cancer is associated with aggressive disease and poor prognosis and deletion of Rac1b in a mouse colorectal cancer model reduces tumourigenesis. We demonstrate that RAC1B interacts with, and is required for efficient activation of the EGFR signalling pathway. Moreover, RAC1B inhibition sensitises cetuximab resistant human tumour organoids to the effects of EGFR inhibition, outlining a potential therapeutic target for improving the clinical efficacy of EGFR inhibitors in colorectal cancer.

Silver nanoparticle polymer composite based humidity sensor.

Silver nanoparticles were synthesised by a chemical reduction process in order to produce an aqueous colloidal dispersion. The resulting colloids were then characterised by a combination of UV-Vis spectroscopy, dynamic light scattering, X-ray diffraction and transmission electron microscopy and the nanoparticles were found to have an average diameter of 20-22 nm. The Ag/polymer nanocomposites were then applied to platinum interdigital electrodes as sensor coatings and the capability of the resulting sensor as a humidity detector investigated. With the application of 1 V, a current developed which was found to be directly proportional to humidity levels. The sensor gives a reversible, selective and rapid response which is proportional to levels of humidity within the range of 10% RH to 60% RH. An investigation into the mechanism of the sensor's response was conducted and the response was found to correlate well with a second order Langmuir adsorption model.

Hydrological response to a seafloor spreading episode on the Juan de Fuca ridge.

Seafloor hydrothermal systems are known to respond to seismic and magmatic activity along mid-ocean ridges, often resulting in locally positive changes in hydrothermal discharge rate, temperature and microbial activity, and shifts in composition occurring at the time of earthquake swarms and axial crustal dike injections. Corresponding regional effects have also been observed. Here we present observations of a hydrological response to seafloor spreading activity, which resulted in a negative formation-fluid pressure transient during and after an earthquake swarm in the sediment-sealed igneous crust of the Middle Valley rift of the northernmost Juan de Fuca ridge. The observations were made with a borehole seal and hydrologic observatory originally established in 1991 to study the steady-state pressure and temperature conditions in this hydrothermally active area. The magnitude of the co-seismic response is consistent with the elastic strain that would be expected from the associated earthquakes, but the prolonged negative pressure transient after the swarm is surprising and suggests net co-seismic dilatation of the upper, permeable igneous crust. The rift valley was visited four weeks after the onset of the seismic activity, but no signature of increased hydrothermal activity was detected in the water column. It appears that water, not magma, filled the void left by this spreading episode.