OX40 and 4-1BB delineate distinct immune profiles in sarcoma.

Systemic relapse after radiotherapy and surgery is the major cause of disease-related mortality in sarcoma patients. Combining radiotherapy and immunotherapy is under investigation as a means to improve response rates. However, the immune contexture of sarcoma is understudied. Here, we use a retrospective cohort of sarcoma patients, treated with neoadjuvant radiotherapy, and TCGA data. We explore therapeutic targets of relevance to sarcoma, using genomics and multispectral immunohistochemistry to provide insights into the tumor immune microenvironment across sarcoma subtypes. Differential gene expression between radioresponsive myxoid liposarcoma (MLPS) and more radioresistant undifferentiated pleomorphic sarcoma (UPS) indicated UPS contained higher transcript levels of a number of immunotherapy targets (CD73/NT5E, CD39/ENTPD1, CD25/IL2RA, and 4-1BB/TNFRSF9). We focused on 4-1BB/TNFRSF9 and other costimulatory molecules. In TCGA data, 4-1BB correlated to an inflamed and exhausted phenotype. OX40/TNFRSF4 and 4-1BB/TNFRSF9 were highly expressed in sarcoma subtypes versus other cancers. Despite OX40 and 4-1BB being described as Treg markers, we identified that they delineate distinct tumor immune profiles. This was true for sarcoma and other cancers. While only a limited number of samples could be analyzed, spatial analysis of OX40 expression identified two diverse phenotypes of OX40+ Tregs, one associated with and one independent of tertiary lymphoid structures (TLSs). Patient stratification is of intense interest for immunotherapies. We provide data supporting the viewpoint that a cohort of sarcoma patients, appropriately selected, are promising candidates for immunotherapies. Spatial profiling of OX40+ Tregs, in relation to TLSs, could be an additional metric to improve future patient stratification.

A multi-species powder dropper for magnetic fusion applications.

We present a device for controlled injection of a variety of materials in powder form. The system implements four independent feeder units, arranged to share a single vertical drop tube. Each unit consists of a 80 ml reservoir, coupled to a horizontal linear trough, where a layer of powder is advanced by piezo-electric agitation at a speed proportional to the applied voltage, until it falls into a drop tube. The dropper has been tested with a number of impurities of low (B, BN, C), intermediate (Si, SiC), and high Z (Sn) and a variety of microscopic structures (flakes, spheres, rocks) and sizes (5-200 µm). For low Z materials, drop rates ∼2-200 mg/s have been obtained showing good repeatability and uniformity. A calibrated light-emitting diode (LED)-based flowmeter allows measuring and monitoring the drop rate during operation. The fast time-response of the four feeders allows combination of steady and pulsed injections, providing a flexible tool for controlled-dose, real-time impurity injection in fusion plasmas.

The SSRI citalopram increases the sensitivity of the human circadian system to light in an acute dose.

RATIONALE: Disturbances of the circadian system are common in depression. Though they typically subside when depression is treated with antidepressants, the mechanism by which this occurs is unknown. Despite being the most commonly prescribed class of antidepressants, the effect of selective serotonin reuptake inhibitors (SSRIs) on the human circadian clock is not well understood. OBJECTIVE: To examine the effect of the SSRI citalopram (30 mg) on the sensitivity of the human circadian system to light. METHODS: This study used a double-blind, placebo-controlled, within-subjects, crossover design. Participants completed two melatonin suppression assessments in room level light (~ 100 lx), taking either a single dose of citalopram 30 mg or a placebo at the beginning of each light exposure. Melatonin suppression was calculated by comparing placebo and citalopram light exposure conditions to a dim light baseline. RESULTS: A 47% increase in melatonin suppression was observed after administration of an acute dose of citalopram, with all participants showing more suppression after citalopram administration (large effect, d = 1.54). Further, melatonin onset occurred later under normal room light with citalopram compared to placebo. CONCLUSIONS: Increased sensitivity of the circadian system to light could assist in explaining some of the inter-individual variability in antidepressant treatment responses, as it is likely to assist in recovery in some patients, while causing further disruption for others.

Upgraded flowing liquid lithium limiter for improving Li coverage uniformity and erosion resistance in EAST device.

We report on design and technology improvements for a flowing liquid lithium (FLiLi) limiter inserted into auxiliary heated discharges in the experimental advanced superconducting tokamak device. In order to enhance Li coverage uniformity and erosion resistance, a new liquid Li distributor with homogenous channels was implemented. In addition, two independent electromagnetic pumps and a new horizontal capillary structure contributed to an improvement in the observed Li flow uniformity (from 30% in the previous FLiLi design to >80% in this FLiLi design). To improve limiter surface erosion resistance, hot isostatic press technology was applied, which improved the thermal contact between thin stainless steel protective layers covering the Cu heat sink. The thickness of the stainless steel layer was increased from 0.1 mm to 0.5 mm, which also helped macroscopic erosion resilience. Despite the high auxiliary heating power up to 4.5 MW, no Li bursts were recorded from FLiLi, underscoring the improved performance of this new design.

Refractive index determination by coherence scanning interferometry.

Coherence scanning interferometry is established as a powerful noncontact, three-dimensional, metrology technique used to determine accurate surface roughness and topography measurements with subnanometer precision. The helical complex field (HCF) function is a topographically defined helix modulated by the electrical field reflectance, originally developed for the measurement of thin films. An approach to extend the capability of the HCF function to determine the spectral refractive index of a substrate or absorbing film has recently been proposed. In this paper, we confirm this new capability, demonstrating it on surfaces of silicon, gold, and a gold/palladium alloy using silica and zirconia oxide thin films. These refractive index dispersion measurements show good agreement with those obtained by spectroscopic ellipsometry.

Oncolytic vaccinia virus as a vector for therapeutic sodium iodide symporter gene therapy in prostate cancer.

Oncolytic strains of vaccinia virus are currently in clinical development with clear evidence of safety and promising signs of efficacy. Addition of therapeutic genes to the viral genome may increase the therapeutic efficacy of vaccinia. We evaluated the therapeutic potential of vaccinia virus expressing the sodium iodide symporter (NIS) in prostate cancer models, combining oncolysis, external beam radiotherapy and NIS-mediated radioiodide therapy. The NIS-expressing vaccinia virus (VV-NIS), GLV-1h153, was tested in in vitro analyzes of viral cell killing, combination with radiotherapy, NIS expression, cellular radioiodide uptake and apoptotic cell death in PC3, DU145, LNCaP and WPMY-1 human prostate cell lines. In vivo experiments were carried out in PC3 xenografts in CD1 nude mice to assess NIS expression and tumor radioiodide uptake. In addition, the therapeutic benefit of radioiodide treatment in combination with viral oncolysis and external beam radiotherapy was measured. In vitro viral cell killing of prostate cancers was dose- and time-dependent and was through apoptotic mechanisms. Importantly, combined virus therapy and iodizing radiation did not adversely affect oncolysis. NIS gene expression in infected cells was functional and mediated uptake of radioiodide both in vitro and in vivo. Therapy experiments with both xenograft and immunocompetent Transgenic Adenocarcinoma of the Mouse Prostate (TRAMP) mouse models showed that the addition of radioiodide to VV-NIS-infected tumors was more effective than each single-agent therapy, restricting tumor growth and increasing survival. In conclusion, VV-NIS is effective in prostate cancer models. This treatment modality would be an attractive complement to existing clinical radiotherapy practice.

New steady-state quiescent high-confinement plasma in an experimental advanced superconducting tokamak.

A critical challenge facing the basic long-pulse high-confinement operation scenario (H mode) for ITER is to control a magnetohydrodynamic (MHD) instability, known as the edge localized mode (ELM), which leads to cyclical high peak heat and particle fluxes at the plasma facing components. A breakthrough is made in the Experimental Advanced Superconducting Tokamak in achieving a new steady-state H mode without the presence of ELMs for a duration exceeding hundreds of energy confinement times, by using a novel technique of continuous real-time injection of a lithium (Li) aerosol into the edge plasma. The steady-state ELM-free H mode is accompanied by a strong edge coherent MHD mode (ECM) at a frequency of 35-40 kHz with a poloidal wavelength of 10.2 cm in the ion diamagnetic drift direction, providing continuous heat and particle exhaust, thus preventing the transient heat deposition on plasma facing components and impurity accumulation in the confined plasma. It is truly remarkable that Li injection appears to promote the growth of the ECM, owing to the increase in Li concentration and hence collisionality at the edge, as predicted by GYRO simulations. This new steady-state ELM-free H-mode regime, enabled by real-time Li injection, may open a new avenue for next-step fusion development.

Methods and preliminary measurement results of liquid Li wettability.

A test of lithium wettability was performed in high vacuum (< 3 × 10(-4) Pa). High magnification images of Li droplets on stainless steel substrates were produced and processed using the MATLAB(®) program to obtain clear image edge points. In contrast to the more standard "θ/2" or polynomial fitting methods, ellipse fitting of the complete Li droplet shape resulted in reliable contact angle measurements over a wide range of contact angles. Using the ellipse fitting method, it was observed that the contact angle of a liquid Li droplet on a stainless steel substrate gradually decreased with increasing substrate temperature. The critical wetting temperature of liquid Li on stainless steel was observed to be about 290 °C.

Sleep difficulty mediates effects of vasomotor symptoms on mood in younger breast cancer survivors.

OBJECTIVE: Treatment-induced early menopause occurs in > 80% of premenopausal women diagnosed with breast cancer. This study explored the relationship between vasomotor symptoms (VMS), sleep and mood in women aged 40-51 years with non-metastatic breast cancer. METHODS: Cross-sectional study using validated questionnaires (Greene Climacteric scale and Hospital Anxiety and Depression Scale, HADS). Women (n = 114) were recruited from the community and hospital outpatient clinics. Frequency determination and structural equation modeling (SEMod) were used to examine the relationship between the latent variables: VMS, anxiety, and depression, and the indicator variable: difficulty sleeping. RESULTS: Participants' mean age was 47 years and 94% became menopausal after breast cancer diagnosis. Difficulty sleeping was reported by 82% of women with 46% reporting (Likert scale) 'quite a bit/extremely'. Most women reported night sweats (77% of women: 47% reporting 'quite a bit/extremely') and hot flushes (84% of women: 50% reporting 'quite a bit/extremely'). HADS scores indicated clinically relevant depression and anxiety in 98% and 99% of women, respectively. SEMod revealed that VMS contributed to difficulty sleeping (standardized coefficient = 0.54; p < 0.001) and difficulty sleeping mediated the relationship between VMS and anxiety (standardized coefficient = 0.34; p = 0.03). However, difficulty sleeping did not have a significant direct impact on depression (standardized coefficient = -0.03; p = 0.8), although anxiety was a strong predictor of depression (standardized coefficient = 0.83; p = 0.015). CONCLUSIONS: VMS, sleep and mood disturbance are commonly experienced by younger women with breast cancer. Using SEMod, we demonstrate for the first time that VMS may directly influence sleep in these women. VMS may have an indirect effect on mood, partly mediated by sleep difficulty.

Synergistic cytotoxicity of radiation and oncolytic Lister strain vaccinia in (V600D/E)BRAF mutant melanoma depends on JNK and TNF-α signaling.

Melanoma is an aggressive skin cancer that carries an extremely poor prognosis when local invasion, nodal spread or systemic metastasis has occurred. Recent advances in melanoma biology have revealed that RAS-RAF-MEK-ERK signaling has a pivotal role in governing disease progression and treatment resistance. Proof-of-concept clinical studies have shown that direct BRAF inhibition yields impressive responses in advanced disease but these are short-lived as treatment resistance rapidly emerges. Therefore, there is a pressing need to develop new targeted strategies for BRAF mutant melanoma. As such, oncolytic viruses represent a promising cancer-specific approach with significant activity in melanoma. This study investigated interactions between genetically-modified vaccinia virus (GLV-1h68) and radiotherapy in melanoma cell lines with BRAF mutant, Ras mutant or wild-type genotype. Preclinical studies revealed that GLV-1h68 combined with radiotherapy significantly increased cytotoxicity and apoptosis relative to either single agent in (V600D)BRAF/(V600E)BRAF mutant melanoma in vitro and in vivo. The mechanism of enhanced cytotoxicity with GLV-1h68/radiation (RT) was independent of viral replication and due to attenuation of JNK, p38 and ERK MAPK phosphorylation specifically in BRAF mutant cells. Further studies showed that JNK pathway inhibition sensitized BRAF mutant cells to GLV-1h68-mediated cell death, mimicking the effect of RT. GLV-1h68 infection activated MAPK signaling in (V600D)BRAF/(V600E)BRAF mutant cell lines and this was associated with TNF-α secretion which, in turn, provided a prosurvival signal. Combination GLV-1h68/RT (or GLV-1h68/JNK inhibition) caused abrogation of TNF-α secretion. These data provide a strong rationale for combining GLV-1h68 with irradiation in (V600D/E)BRAF mutant tumors.

Oncolytic Vaccinia virus and radiotherapy in head and neck cancer.

OBJECTIVE: Oncolytic forms of attenuated Vaccinia virus are now in clinical development, assessing the compatibility of this novel treatment with radiotherapy may reveal exploitable synergistic relationships. MATERIALS AND METHODS: In vitro analyses of cell killing, cell cycle effects and caspase activation were carried out on HN3, HN5, CAL27, Detroit, SIHN5B, and PJ41 cells. In vivo studies of the virus and X-radiation were performed on H&N xenografts in CD1 nude mice. RESULTS: Cell killing in vitro was demonstrated to be dose- and time-dependent. Infection causes an increase in S-phase and sub-G1 cells. A dose dependent increase in active caspase-3 indicated induction of apoptosis. Xenografts injected with Vaccinia stabilised and frequently completely regressed. Combination with radiation generated additional cell death, induction of caspase activity and in vivo further improved long term regression rates. CONCLUSIONS: These data support continued exploration of this therapy combination and indicates potential for clinical trials in head and neck cancer.

Synergistic cytotoxicity of oncolytic reovirus in combination with cisplatin-paclitaxel doublet chemotherapy.

Oncolytic reovirus is currently under active investigation in a range of tumour types. Early phase studies have shown that this agent has modest monotherapy efficacy and its future development is likely to focus on combination regimens with cytotoxic chemotherapy. Indeed, phase I/II clinical trials have confirmed that reovirus can be safely combined with cytotoxic drugs, including a platin-taxane doublet regimen, which is currently being tested in a phase III clinical trial in patients with relapsed/metastatic head and neck cancer. Therefore, we have tested this triple (reovirus, cisplatin, paclitaxel) combination therapy in a panel of four head and neck cancer cell lines. Using the combination index (CI) method, the triple therapy demonstrated synergistic cytotoxicity in vitro in both malignant and non-malignant cell lines. In head and neck cancer cell lines, this was associated with enhanced caspase 3 and 7 cleavage, but no increase in viral replication. In vitro analyses confirmed colocalisation of markers of reovirus infection and caspase 3. Triple therapy was significantly more effective than reovirus or cisplatin-paclitaxel in athymic nude mice. These data suggest that the combination of reovirus plus platin-taxane doublet chemotherapy has significant activity in head and neck cancer and underpin the current phase III study in this indication.

The importance of informed consent in patients with wet age-related macular degeneration considering intravitreal anti-vascular endothelial growth factor treatments.

Retinal pigment epithelial (RPE) tears are now a documented potential complication following the intravitreal injection of anti-vascular endothelial growth factor (VEGF) treatments for neovascular age-related macular degeneration. Patients are often not well consented regarding this risk and thus we retrospectively analyzed the data from all of our patients undergoing this treatment over a six month period. Our findings highlighted the fact that the three patients (out of thirty) who had developed this RPE tear complication were initially all diagnosed with a pigment epithelial detachment (which is a type of macular degeneration in question). Therefore, we have adjusted our informed consent procedure such that all patients with "wet" macular degeneration and especially those with pigment epithelial detachments are now fully consented regarding the risks of the intravitreal treatment, which could potentially damage their vision further.

Continuous improvement of H-mode discharge performance with progressively increasing lithium coatings in the National Spherical Torus Experiment.

Lithium wall coatings have been shown to reduce recycling, improve energy confinement, and suppress edge localized modes in the National Spherical Torus Experiment. Here, we show that these effects depend continuously on the amount of predischarge lithium evaporation. We observed a nearly monotonic reduction in recycling, decrease in electron transport, and modification of the edge profiles and stability with increasing lithium. These correlations challenge basic expectations, given that even the smallest coatings exceeded that needed for a nominal thickness of the order of the implantation range.

Preliminary results from the Alcator C-Mod polarimeter.

A poloidally viewing far infrared polarimeter diagnostic is being developed for the Alcator C-Mod tokamak, and will be used to determine the q-profile and to study density and magnetic field fluctuations. A three-chord version of what will eventually be up to a ten-chord system has been designed and fabricated and will be installed on C-Mod before the end of the current run period. Bench tests of a single chord mock-up of this system show acceptable noise levels for the planned measurements. We will discuss the analysis and experimental techniques used to diagnose and reduce noise sources.

Experimental setup for tungsten transport studies at the NSTX tokamak.

Tungsten particles have been introduced into the National Spherical Torus Experiment (NSTX) in Princeton with the purpose to investigate the effects of tungsten injection on subsequent plasma discharges. An experimental setup for the study of tungsten particle transport is described where the particles are introduced into the tokamak using a modified particle dropper, otherwise used for lithium-powder injection. An initial test employing a grazing-incidence extreme ultraviolet spectrometer demonstrates that the tungsten-transport setup could serve to infer particle transport from the edge to the hot central plasmas of NSTX.

The precautionary principle: what is the risk of reusing disposable drops in routine ophthalmology consultations and what are the costs of reducing this risk to zero?

BACKGROUND: Instilling eye drops is a ubiquitous procedure in eye clinics. This audit aimed to assess the risk of contamination of disposable droppers and to quantify the financial and waste implications of reducing this risk to zero by using disposable droppers only once. METHODS: A total of 100 disposable Minims were used to place one drop in each eye of 70 patients. The dropper tip was then cultured for aerobic and anaerobic microbes. RESULTS: Coagulase-negative staphylococcus was cultured from five samples. The contamination rate per drop application was 2.5%. The risk of cross-contamination with coagulase-negative staphylococcus would be between 1 : 400 and 1 : 80 if the bottle was reused once or six times. Reducing this risk to zero costs between pound2.75 and pound4.6 million per annum and generates between 6.85 and 11.42 more tonnes of paper waste and between 12.69 and 21.15 more tonnes of plastic waste than a strategy that reuses the disposable dropper. CONCLUSION: Reducing the risk of dropper contamination and subsequent cross infection has financial and environmental costs. As exposure to coagulase-negative staphylococcus is not necessarily associated with infection, it would be useful to decide acceptable risk levels for a given cost to maximise both cost-effectiveness and patient safety.

Edge-localized-mode suppression through density-profile modification with lithium-wall coatings in the National Spherical Torus Experiment.

Reduction or elimination of edge localized modes (ELMs) while maintaining high confinement is essential for future fusion devices, e.g., the ITER. An ELM-free regime was recently obtained in the National Spherical Torus Experiment, following lithium (Li) evaporation onto the plasma-facing components. Edge stability calculations indicate that the pre-Li discharges were unstable to low-n peeling or ballooning modes, while broader pressure profiles stabilized the post-Li discharges. Normalized energy confinement increased by 50% post Li, with no sign of ELMs up to the global stability limit.