Redirecting raltitrexed from cancer cell thymidylate synthase to Mycobacterium tuberculosis phosphopantetheinyl transferase.

There is a compelling need to find drugs active against Mycobacterium tuberculosis (Mtb). 4'-Phosphopantetheinyl transferase (PptT) is an essential enzyme in Mtb that has attracted interest as a potential drug target. We optimized a PptT assay, used it to screen 422,740 compounds, and identified raltitrexed, an antineoplastic antimetabolite, as the most potent PptT inhibitor yet reported. While trying unsuccessfully to improve raltitrexed's ability to kill Mtb and remove its ability to kill human cells, we learned three lessons that may help others developing antibiotics. First, binding of raltitrexed substantially changed the configuration of the PptT active site, complicating molecular modeling of analogs based on the unliganded crystal structure or the structure of cocrystals with inhibitors of another class. Second, minor changes in the raltitrexed molecule changed its target in Mtb from PptT to dihydrofolate reductase (DHFR). Third, the structure-activity relationship for over 800 raltitrexed analogs only became interpretable when we quantified and characterized the compounds' intrabacterial accumulation and transformation.

The Remove-the-Mask Open-Source head and neck Surface-Guided radiation therapy system.

Background and Purpose: Surface Guided Radiotherapy (SGRT) for head and neck radiotherapy is challenging as obstructions are common and non-rigid facial motion can compromise surface accuracy. The purpose of this work was to develop and benchmark the Remove the Mask (RtM) SGRT system, an open-source system especially designed to address the challenges faced in radiotherapy of head and neck cancer. Materials and Methods: The accuracy of the RtM SGRT system was benchmarked using a head phantom positioned on a robotic motion platform capable of sub-millimetre accuracy which was used to induce unidirectional shifts and to reproduce three real head motion traces. We also assessed the accuracy of the system in ten humans volunteers. The ground truth motion of the volunteers was obtained using a commercial motion capture system with an accuracy < 0.3 mm. Results: The mean tracking error of the RtM SGRT system for the ten volunteers was of -0.1 ± 0.4 mm -0.6 ± 0.6 mm and 0.3 ± 0.2 mm, and 0.0 ± 0.2° 0.0 ± 0.1° and 0.0 ± 0.2° for translations and rotations along the left-right, superior-inferior and anterior-posterior axes respectively and we also found similar results in measurements with the head phantom. Forced facial motion was associated with lower tracking accuracy. The RtM SGRT system achieved submillimetre accuracy. Conclusion: The RtM SGRT system is a low-cost, easy to build and open-source SGRT system that can achieve an accuracy that meets international commissioning guidelines. Its open-source and modular design allows for the development and easy translation of novel surface tracking techniques.

A kinematics-based method for creating deformed patient-derived head and neck CT scans.

CT scans of the head and neck have multiple clinical uses, and simulating deformation of these CT scans allows for predicting patient motion and data augmentation for machine-learning methods. Current methods for creating patient-derived deformed CT scans require multiple scans or use unrealistic head and neck motion. This paper describes the CTHeadDeformation software package which allows for realistic synthetic deformation of head and neck CT scans for small amounts of motion. CTHeadDeformation is a python-based package that uses a kinematics-based approach using anatomical landmarks, and rigid/non-rigid registration to create a realistic patient-derived deformed CT scan. CTHeadDeformation is also designed for simple clinical implementation. The CTHeadDeformation software package was demonstrated on a head and neck CT scan of one patient. The CT scan was deformed in the anterior-posterior, superior-inferior, and left-right directions. Internal organ motion and more complex combination motions were also simulated. The results showed the patient's CT scan was able to be deformed in a way that preserved the shape and location of the anatomy.Clinical Relevance- This method allows for the realistic simulation of head and neck motion in CT scans. Clinical applications including simulating how patient motion affects radiation therapy treatment effectiveness. The CTHeadDeformation software can also be used to train machine-learning networks that are robust to patient motion, or to generate ground truth images for imaging or segmentation grand challenges.

Nano X Image Guidance in radiation therapy: feasibility study protocol for cone beam computed tomography imaging with gravity-induced motion.

BACKGROUND: This paper describes the protocol for the Nano X Image Guidance (Nano X IG) trial, a single-institution, clinical imaging study. The Nano X is a prototype fixed-beam radiotherapy system developed to investigate the feasibility of a low-cost, compact radiotherapy system to increase global access to radiation therapy. This study aims to assess the feasibility of volumetric image guidance with cone beam computed tomography (CBCT) acquired during horizontal patient rotation on the Nano X radiotherapy system. METHODS: In the Nano X IG study, we will determine whether radiotherapy image guidance can be performed with the Nano X radiotherapy system where the patient is horizontally rotated while scan projections are acquired. We will acquire both conventional CBCT scans and Nano X CBCT scans for 30 patients aged 18 and above and receiving radiotherapy for head/neck or upper abdomen cancers. For each patient, a panel of experts will assess the image quality of Nano X CBCT scans against conventional CBCT scans. Each patient will receive two Nano X CBCT scans to determine the image quality reproducibility, the extent and reproducibility of patient motion and assess patient tolerance. DISCUSSION: Fixed-beam radiotherapy systems have the potential to help ease the current shortfall and increase global access to radiotherapy treatment. Advances in image guidance could facilitate fixed-beam radiotherapy using horizontal patient rotation. The efficacy of this radiotherapy approach is dependent on our ability to image and adapt to motion due to rotation and for patients to tolerate rotation during treatment. TRIAL REGISTRATION: ClinicalTrials.gov, NCT04488224. Registered on 27 July 2020.

Realistic CT data augmentation for accurate deep-learning based segmentation of head and neck tumors in kV images acquired during radiation therapy.

BACKGROUND: Using radiation therapy (RT) to treat head and neck (H&N) cancers requires precise targeting of the tumor to avoid damaging the surrounding healthy organs. Immobilisation masks and planning target volume margins are used to attempt to mitigate patient motion during treatment, however patient motion can still occur. Patient motion during RT can lead to decreased treatment effectiveness and a higher chance of treatment related side effects. Tracking tumor motion would enable motion compensation during RT, leading to more accurate dose delivery. PURPOSE: The purpose of this paper is to develop a method to detect and segment the tumor in kV images acquired during RT. Unlike previous tumor segmentation methods for kV images, in this paper, a process for generating realistic and synthetic CT deformations was developed to augment the training data and make the segmentation method robust to patient motion. Detecting the tumor in 2D kV images is a necessary step toward 3D tracking of the tumor position during treatment. METHOD: In this paper, a conditional generative adversarial network (cGAN) is presented that can detect and segment the gross tumor volume (GTV) in kV images acquired during H&N RT. Retrospective data from 15 H&N cancer patients obtained from the Cancer Imaging Archive were used to train and test patient-specific cGANs. The training data consisted of digitally reconstructed radiographs (DRRs) generated from each patient's planning CT and contoured GTV. Training data was augmented by using synthetically deformed CTs to generate additional DRRs (in total 39 600 DRRs per patient or 25 200 DRRs for nasopharyngeal patients) containing realistic patient motion. The method for deforming the CTs was a novel deformation method based on simulating head rotation and internal tumor motion. The testing dataset consisted of 1080 DRRs for each patient, obtained by deforming the planning CT and GTV at different magnitudes to the training data. The accuracy of the generated segmentations was evaluated by measuring the segmentation centroid error, Dice similarity coefficient (DSC) and mean surface distance (MSD). This paper evaluated the hypothesis that when patient motion occurs, using a cGAN to segment the GTV would create a more accurate segmentation than no-tracking segmentations from the original contoured GTV, the current standard-of-care. This hypothesis was tested using the 1-tailed Mann-Whitney U-test. RESULTS: The magnitude of our cGAN segmentation centroid error was (mean ± standard deviation) 1.1 ± 0.8 mm and the DSC and MSD values were 0.90 ± 0.03 and 1.6 ± 0.5 mm, respectively. Our cGAN segmentation method reduced the segmentation centroid error (p < 0.001), and MSD (p = 0.031) when compared to the no-tracking segmentation, but did not significantly increase the DSC (p = 0.294). CONCLUSIONS: The accuracy of our cGAN segmentation method demonstrates the feasibility of this method for H&N cancer patients during RT. Accurate tumor segmentation of H&N tumors would allow for intrafraction monitoring methods to compensate for tumor motion during treatment, ensuring more accurate dose delivery and enabling better H&N cancer patient outcomes.

Perspective on Schistosomiasis Drug Discovery: Highlights from a Schistosomiasis Drug Discovery Workshop at Wellcome Collection, London, September 2022.

In September 2022, the Drug Discovery Unit at the University of Dundee, UK, organised an international meeting at the Wellcome Collection in London to explore the current clinical situation and challenges associated with treating schistosomiasis. The aim of this meeting was to discuss the need for new treatments in view of the clinical situation and to ascertain what the key requirements would be for any potential new anti-schistosomals. This information will be essential to inform ongoing drug discovery efforts for schistosomiasis. We also discussed the potential drug discovery pathway and associated criteria for progressing compounds to the clinic. To date, praziquantel (PZQ) is the only drug available to treat all species causing schistosomiasis, but it is often unable to completely clear parasites from an infected patient, partially due to its inactivity against juvenile worms. PZQ-mediated mass drug administration campaigns conducted in endemic areas (e.g., sub-Saharan Africa, where schistosomiasis is primarily prevalent) have contributed to reducing the burden of disease but will not eliminate the disease as a public health problem. The potential for Schistosoma to develop resistance towards PZQ, as the sole treatment available, could become a concern. Consequently, new anthelmintic medications are urgently needed, and this Perspective aims to capture some of the learnings from our discussions on the key criteria for new treatments.

Spontaneous perspective-taking and its relation to schizotypy.

INTRODUCTION: Patients with schizophrenia differ from healthy controls in the extent that they spontaneously take another's perspective. For such effects, it is difficult to separate the influence of schizophrenia from multiple potential confounders. Here, for the first time, associations between spontaneous perspective-taking and schizotypy were investigated in a nonclinical population. METHODS: Adult participants completed both a Schizotypal Personality Questionnaire (SPQ-BRU) and a novel online adaptation of a visual perspective-taking task that required participants to make judgements both from their own perspective and that of a human avatar. RESULTS: Response times were elevated when the avatar's perspective was inconsistent with that of the participant, providing evidence of spontaneous perspective-taking. This demonstrates that the visual perspective-taking task can be successfully implemented in an online format. However, schizotypy did not predict these spontaneous perspective-taking effects. CONCLUSIONS: Unlike explicit mentalising, this form of implicit mentalising is not affected by nonclinical manifestations of schizotypy traits. This implies that impairment of general neurocognitive function contributes to altered spontaneous perspective-taking in schizophrenia. A novel account based on the cognitive control processes involved in perspective selection and the role of attention in perspective calculation reconciles apparently contradictory findings of earlier studies comparing patients with schizophrenia and healthy controls.

A Wearable Ballistocardiography Device for Estimating Heart Rate During Positive Airway Pressure Therapy: Investigational Study Among the General Population.

BACKGROUND: Obstructive sleep apnea (OSA) is a condition in which a person's airway is obstructed during sleep, thus disturbing their sleep. People with OSA are at a higher risk of developing heart problems. OSA is commonly treated with a positive airway pressure (PAP) therapy device, which is used during sleep. The PAP therapy setup provides a good opportunity to monitor the heart health of people with OSA, but no simple, low-cost method is available for the PAP therapy device to monitor heart rate (HR). OBJECTIVE: This study aims to develop a simple, low-cost device to monitor the HR of people with OSA during PAP therapy. This device was then tested on a small group of participants to investigate the feasibility of the device. METHODS: A low-cost and simple device to monitor HR was created by attaching a gyroscope to a PAP mask, thus integrating HR monitoring into PAP therapy. The gyroscope signals were then analyzed to detect heartbeats, and a Kalman filter was used to produce a more accurate and consistent HR signal. In this study, 19 participants wore the modified PAP mask while the mask was connected to a PAP device. Participants lay in 3 common sleeping positions and then underwent 2 different PAP therapy modes to determine if these affected the accuracy of the HR estimation. RESULTS: Before the PAP device was turned on, the median HR error was <5 beats per minute, although the HR estimation error increased when participants lay on their side compared with when participants lay on their back. Using the different PAP therapy modes did not significantly increase the HR error. CONCLUSIONS: These results show that monitoring HR from gyroscope signals in a PAP mask is possible during PAP therapy for different sleeping positions and PAP therapy modes, suggesting that long-term HR monitoring of OSA during PAP therapy may be possible.

The adaptation and investigation of cone-beam CT reconstruction algorithms for horizontal rotation fixed-gantry scans of rabbits.

Fixed-gantry radiation therapy has been proposed as a low-cost alternative to the conventional rotating-gantry radiation therapy, that may help meet the rising global treatment demand. Fixed-gantry systems require gravitational motion compensated reconstruction algorithms to produce cone-beam CT (CBCT) images of sufficient quality for image guidance. The aim of this work was to adapt and investigate five CBCT reconstruction algorithms for fixed-gantry CBCT images. The five algorithms investigated were Feldkamp-Davis-Kress (FDK), prior image constrained compressed sensing (PICCS), gravitational motion compensated FDK (GMCFDK), motion compensated PICCS (MCPICCS) (a novel CBCT reconstruction algorithm) and simultaneous motion estimation and iterative reconstruction (SMEIR). Fixed-gantry and rotating-gantry CBCT scans were acquired of 3 rabbits, with the rotating-gantry scans used as a reference. Projections were sorted into rotation bins, based on the angle of rotation of the rabbit during image acquisition. The algorithms were compared using the structural similarity index measure root mean square error, and reconstruction time. Evaluation of the reconstructed volumes showed that, when compared with the reference rotating-gantry volume, the conventional FDK algorithm did not accurately reconstruct fixed-gantry CBCT scans. Whilst the PICCS reconstruction algorithm reduced some motion artefacts, the motion estimation reconstruction methods (GMCFDK, MCPICCS and SMEIR) were able to greatly reduce the effect of motion artefacts on the reconstructed volumes. This finding was verified quantitatively, with GMCFDK, MCPICCS and SMEIR reconstructions having RMSE 17%-19% lower and SSIM 1% higher than a conventional FDK. However, all motion compensated fixed-gantry CBCT reconstructions had a 56%-61% higher RMSE and 1.5% lower SSIM than FDK reconstructions of conventional rotating-gantry CBCT scans. The results show that motion compensation is required to reduce motion artefacts for fixed-gantry CBCT reconstructions. This paper further demonstrates the feasibility of fixed-gantry CBCT scans, and the ability of CBCT reconstruction algorithms to compensate for motion due to horizontal rotation.

Pre-treatment and real-time image guidance for a fixed-beam radiotherapy system.

PURPOSE: A radiotherapy system with a fixed treatment beam and a rotating patient positioning system could be smaller, more robust and more cost effective compared to conventional rotating gantry systems. However, patient rotation could cause anatomical deformation and compromise treatment delivery. In this work, we demonstrate an image-guided treatment workflow with a fixed beam prototype system that accounts for deformation during rotation to maintain dosimetric accuracy. METHODS: The prototype system consists of an Elekta Synergy linac with the therapy beam orientated downward and a custom-built patient rotation system (PRS). A phantom that deforms with rotation was constructed and rotated within the PRS to quantify the performance of two image guidance techniques: motion compensated cone-beam CT (CBCT) for pre-treatment volumetric imaging and kilovoltage infraction monitoring (KIM) for real-time image guidance. The phantom was irradiated with a 3D conformal beam to evaluate the dosimetric accuracy of the workflow. RESULTS: The motion compensated CBCT was used to verify pre-treatment position and the average calculated position was within -0.3 ± 1.1 mm of the phantom's ground truth position at 0°. KIM tracked the position of the target in real-time as the phantom was rotated and the average calculated position was within -0.2 ± 0.8 mm of the phantom's ground truth position. A 3D conformal treatment delivered on the prototype system with image guidance had a 3%/2 mm gamma pass rate of 96.3% compared to 98.6% delivered using a conventional rotating gantry linac. CONCLUSIONS: In this work, we have shown that image guidance can be used with fixed-beam treatment systems to measure and account for changes in target position in order to maintain dosimetric coverage during horizontal rotation. This treatment modality could provide a viable treatment option when there insufficient space for a conventional linear accelerator or where the cost is prohibitive.

Dehydration in older people: A systematic review of the effects of dehydration on health outcomes, healthcare costs and cognitive performance.

OBJECTIVE: To systematically examine the effect of dehydration on health outcomes, identify associated financial costs and consider impacts on cognitive performance in older adults. DESIGN: A systematic review of English-language articles via OVID using MEDLINE, PsychINFO, EMBASE, and others, to March 2018. Included studies examined the relationship between hydration status and health, care costs or cognitive outcome. SETTING: Cross sectional and cohort data from studies reporting on dehydration in older adults. PARTICIPANTS: Adults aged 60 years and older. MEASUREMENTS: Independent quality ratings were assessed for all extracted articles. RESULTS: Of 1684 articles screened, 18 papers (N = 33,707) met inclusion criteria. Participants were recruited from hospital settings, medical long-term care centres and the community dwelling population. Data were synthesised using a narrative summary. Mortality rates were higher in dehydrated patients. Furthermore, health outcomes, including frailty, bradyarrhythmia, transient ischemic attacks, oral health and surgery recovery are linked to and worsened by dehydration. Length of hospital stay, either as a principal or secondary diagnosis, is greater in those with dehydration, compared to those who are euhydrated. Finally, neurocognitive functioning may be impacted by dehydration. There are issues with study design, inconsistency in hydration status measurement and different measures used for outcome assessment. CONCLUSION: Dehydration in older people is associated with increased mortality, poorer course of illness and increased costs for health services. In addition, there is some, but sparse evidence that dehydration in older people is linked to poorer cognitive performance. Intervention studies should test strategies for reducing dehydration in older adults.

At what stage in the drinking process does drinking water affect attention and memory? Effects of mouth rinsing and mouth drying in adults.

Drinking water is important for health and there is an agreement that drinking water facilitates certain cognitive processes. However, the mechanism underlying the effect of drinking water on cognition is unknown. While attention performance is improved by even a very small drink, memory performance seems to require larger drinks for performance enhancement. This suggests that attention could be affected earlier in the drinking process than memory. We aimed to elucidate further the mechanism involved by investigating the stage during the drinking process influencing performance on cognitive tasks. To this end, we compared mouth rinsing and mouth drying. Mouth rinsing was expected to result in improved attention performance and would suggest that the mechanism responsible is located in the mouth and occurs early in the drinking process, before swallowing. Eighty-seven adults participated in either a treatment (mouth rinsing or mouth drying) or control (no intervention) condition. They were assessed at baseline and 20 min later after intervention on measures of visual attention, short-term memory, subjective thirst and mood. Our results showed that mouth rinsing improved visual attention, but not short-term memory, mood or subjective thirst. Mouth drying did not affect performance. Our results support the hypothesis that different mechanisms underlie the effect of drinking water on different cognitive processes. They suggest that merely sipping water, as opposed to having a large drink, can improve attention.

Towards automated in vivo tracheal mucociliary transport measurement: Detecting and tracking particle movement in synchrotron phase-contrast x-ray images.

Accurate in vivo quantification of airway mucociliary transport (MCT) in animal models is important for understanding diseases such as cystic fibrosis, as well as for developing therapies. A non-invasive method of measuring MCT behaviour, based on tracking the position of micron sized particles using synchrotron x-ray imaging, has previously been described. In previous studies, the location (and path) of each particle was tracked manually, which is a time consuming and subjective process. Here we describe particle tracking methods that were developed to reduce the need for manual particle tracking. The MCT marker particles were detected in the synchrotron x-ray images using cascade classifiers. The particle trajectories along the airway surface were generated by linking the detected locations between frames using a modified particle linking algorithm. The developed methods were compared with the manual tracking method on simulated x-ray images, as well as on in vivo images of rat airways acquired at the SPring-8 Synchrotron. The results for the simulated and in vivo images showed that the semi-automatic algorithm reduced the time required for particle tracking when compared with the manual tracking method, and was able to detect MCT marker particle locations and measure particle speeds more accurately than the manual tracking method. Future work will examine the modification of methods to improve particle detection and particle linking algorithms to allow for more accurate fully-automatic particle tracking.

Development and analytical validation of a next-generation sequencing based microsatellite instability (MSI) assay.

BACKGROUND: We have developed and analytically validated a next-generation sequencing (NGS) assay to classify microsatellite instability (MSI) in formalin-fixed paraffin-embedded (FFPE) tumor specimens. METHODOLOGY: The assay relies on DNA-seq evaluation of insertion/deletion (indel) variability at 29 highly informative genomic loci to estimate MSI status without the requirement for matched-normal tissue. The assay has a clinically relevant five-day turnaround time and can be conducted on as little as 20 ng genomic DNA with a batch size of up to forty samples in a single run. RESULTS: The MSI detection method was developed on a training set (n = 94) consisting of 22 MSI-H, 24 MSS, and 47 matched normal samples and tested on an independent test set of 24 MSI-H and 24 MSS specimens. Assay performance with respect to accuracy, reproducibility, precision as well as control sample performance was estimated across a wide range of FFPE samples of multiple histologies to address pre-analytical variability (percent tumor nuclei), and analytical variability (batch size, run, day, operator). Analytical precision studies demonstrated that the assay is highly reproducible and accurate as compared to established gold standard PCR methodology and has been validated through NYS CLEP. SIGNIFICANCE: This assay provides clinicians with robust and reproducible NGS-based MSI testing without the need of matched normal tissue to inform clinical decision making for patients with solid tumors.

Oncologist uptake of comprehensive genomic profile guided targeted therapy.

We describe the extent to which comprehensive genomic profiling (CGP) results were used by oncologists to guide targeted therapy selection in a cohort of solid tumor patients tested as part of standard care at Roswell Park Comprehensive Cancer Center June 2016-June 2017, with adequate follow up through September 2018 (n = 620). Overall, 28.4% of CGP tests advised physicians about targeted therapy use supported by companion diagnostic or practice guideline evidence. Post-test targeted therapy uptake was highest for patients in active treatment at the time of order (86% versus 76% of treatment naïve patients), but also took longer to initiate (median 50 days versus 7 days for treatment naïve patients), with few patients (2.6%) receiving targeted agents prior to testing. 100% of patients with resistance variants did not receive targeted agents. Treatment naïve patients received immunotherapy as the most common alternative. When targeted therapy given off-label or in a trial was the best CGP option, (7%) of patients received it. Our data illustrate the appropriate and heterogeneous use of CGP by oncologists as a longitudinal treatment decision tool based on patient history and treatment needs, and that some patients may benefit from testing prior to initiation of other standard treatments.

Particle coating alters mucociliary transit in excised rat trachea: A synchrotron X-ray imaging study.

We have previously developed non-invasive in vivo mucociliary transport (MCT) monitoring methods using synchrotron phase contrast X-ray imaging (PCXI) to evaluate potential therapies for cystic fibrosis (CF). However, previous in vivo measurements of MCT velocity using this method were lower than those from alternate methods. We hypothesise this was due to the surface chemistry of the uncoated particles. We investigated the effect of particle surface coating on MCT marker performance by measuring the velocity of uncoated, positively-charged (aminated; NH2), and negatively-charged (carboxylated; COOH) particles. The effect of aerosolised hypertonic saline (HS) was also investigated, as previous in vivo measurements showed HS significantly increased MCT rate. PCXI experiments were performed using an ex vivo rat tracheal imaging setup. Prior to aerosol delivery there was little movement of the uncoated particles, whilst the NH2 and COOH particles moved with MCT rates similar to those previously reported. After application of HS the uncoated and COOH particle velocity increased and NH2 decreased. This experiment validated the use of COOH particles as MCT marker particles over the uncoated and NH2 coated particles. Our results suggest that future experiments measuring MCT using synchrotron PCXI should use COOH coated marker particles for more accurate MCT quantification.

Proliferative potential and resistance to immune checkpoint blockade in lung cancer patients.

BACKGROUND: Resistance to immune checkpoint inhibitors (ICIs) has been linked to local immunosuppression independent of major ICI targets (e.g., PD-1). Clinical experience with response prediction based on PD-L1 expression suggests that other factors influence sensitivity to ICIs in non-small cell lung cancer (NSCLC) patients. METHODS: Tumor specimens from 120 NSCLC patients from 10 institutions were evaluated for PD-L1 expression by immunohistochemistry, and global proliferative profile by targeted RNA-seq. RESULTS: Cell proliferation, derived from the mean expression of 10 proliferation-associated genes (namely BUB1, CCNB2, CDK1, CDKN3, FOXM1, KIAA0101, MAD2L1, MELK, MKI67, and TOP2A), was identified as a marker of response to ICIs in NSCLC. Poorly, moderately, and highly proliferative tumors were somewhat equally represented in NSCLC, with tumors with the highest PD-L1 expression being more frequently moderately proliferative as compared to lesser levels of PD-L1 expression. Proliferation status had an impact on survival in patients with both PD-L1 positive and negative tumors. There was a significant survival advantage for moderately proliferative tumors compared to their combined highly/poorly counterparts (p = 0.021). Moderately proliferative PD-L1 positive tumors had a median survival of 14.6 months that was almost twice that of PD-L1 negative highly/poorly proliferative at 7.6 months (p = 0.028). Median survival in moderately proliferative PD-L1 negative tumors at 12.6 months was comparable to that of highly/poorly proliferative PD-L1 positive tumors at 11.5 months, but in both instances less than that of moderately proliferative PD-L1 positive tumors. Similar to survival, proliferation status has impact on disease control (DC) in patients with both PD-L1 positive and negative tumors. Patients with moderately versus those with poorly or highly proliferative tumors have a superior DC rate when combined with any classification schema used to score PD-L1 as a positive result (i.e., TPS ≥ 50% or ≥ 1%), and best displayed by a DC rate for moderately proliferative tumors of no less than 40% for any classification of PD-L1 as a negative result. While there is an over representation of moderately proliferative tumors as PD-L1 expression increases this does not account for the improved survival or higher disease control rates seen in PD-L1 negative tumors. CONCLUSIONS: Cell proliferation is potentially a new biomarker of response to ICIs in NSCLC and is applicable to PD-L1 negative tumors.

Next generation sequencing of PD-L1 for predicting response to immune checkpoint inhibitors.

BACKGROUND: PD-L1 immunohistochemistry (IHC) has been traditionally used for predicting clinical responses to immune checkpoint inhibitors (ICIs). However, there are at least 4 different assays and antibodies used for PD-L1 IHC, each developed with a different ICI. We set to test if next generation RNA sequencing (RNA-seq) is a robust method to determine PD-L1 mRNA expression levels and furthermore, efficacy of predicting response to ICIs as compared to routinely used, standardized IHC procedures. METHODS: A total of 209 cancer patients treated on-label by FDA-approved ICIs, with evaluable responses were assessed for PD-L1 expression by RNA-seq and IHC, based on tumor proportion score (TPS) and immune cell staining (ICS). A subset of serially diluted cases was evaluated for RNA-seq assay performance across a broad range of PD-L1 expression levels. RESULTS: Assessment of PD-L1 mRNA levels by RNA-seq demonstrated robust linearity across high and low expression ranges. PD-L1 mRNA levels assessed by RNA-seq and IHC (TPS and ICS) were highly correlated (p < 2e-16). Sub-analyses showed sustained correlation when IHC results were classified as high or low by clinically accepted cut-offs (p < 0.01), and results did not differ by tumor type or anti-PD-L1 antibody used. Overall, a combined positive PD-L1 result (≥1% IHC TPS and high PD-L1 expression by RNA-Seq) was associated with a 2-to-5-fold higher overall response rate (ORR) compared to a double negative result. Standard assessments of sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) showed that a PD-L1 positive assessment for melanoma samples by RNA-seq had the lowest sensitivity (25%) but the highest PPV (72.7%). Among the three tumor types analyzed in this study, the only non-overlapping confidence interval for predicting response was for "RNA-seq low vs high" in melanoma. CONCLUSIONS: Measurement of PD-L1 mRNA expression by RNA-seq is comparable to PD-L1 expression by IHC both analytically and clinically in predicting ICI response. RNA-seq has the added advantages of being amenable to standardization and avoidance of interpretation bias. PD-L1 by RNA-seq needs to be validated in future prospective ICI clinical studies across multiple histologies.

A Modified Mask for Continuous Cardiac Monitoring during Positive Airway Pressure Therapy.

A Positive Airway Pressure mask was modified to be able to measure the heart rate of the wearer during Positive Airway Pressure (PAP) therapy. The mask was modified by attaching sensors to measure Electrocardiography (ECG) and Photoplethysmography (PPG) signals, from which a heart rate was extracted. The ECG signal was recorded using stainless steel electrodes positioned on the wearer's head and neck. The PPG signal was measured from the wearer's forehead using a reflectance pulse oximeter attached to the mask. The modified device was tested on 19 healthy participants (mean age30.5 ± 9 years) in a simulated sleeping environment. For both the ECG and PPG signals, the device was able to correctly identify more than 95% of heartbeats for the majority of the participants. However, it was found that by combining the ECG and PPG heartbeat data, the percentage of heartbeats that could be detected was increased to 99%. The results showed that it may be possible to use this device to monitor sleep apnoea patients during PAP therapy in their homes.

How does drinking water affect attention and memory? The effect of mouth rinsing and mouth drying on children's performance.

There is general consensus that drinking water facilitates certain cognitive processes. However, it is not yet known what mechanism underlies the effect of drinking on performance and these may be different for different cognitive processes. We sought to elucidate the mechanisms involved by establishing at what stage of the drinking process cognitive performance is influenced. We examined the effect of mouth rinsing and mouth drying on subjective thirst and mood, visual attention and short term memory in children. Data are reported from 24 children aged 9- to 10-years. Children's performance was assessed in three conditions - mouth drying, mouth rinsing and a control (no intervention). In each condition they were assessed twice - at baseline, before intervention, and 20 min later at test. Mouth rinsing improved visual attention performance, but not short term memory, mood or subjective thirst. The effects of mouth drying were more equivocal. The selective nature of the results is consistent with suggestions that different domains of cognition are influenced by different mechanisms.