Spa-RQ: an Image Analysis Tool to Visualise and Quantify Spatial Phenotypes Applied to Non-Small Cell Lung Cancer

To facilitate analysis of spatial tissue phenotypes, we created an open-source tool package named 'Spa-RQ' for 'Spatial tissue analysis: image Registration & Quantification'. Spa-RQ contains software for image registration (Spa-R) and quantitative analysis of DAB staining overlap (Spa-Q). It provides an easy-to-implement workflow for serial sectioning and staining as an alternative to multiplexed techniques. To demonstrate Spa-RQ's applicability, we analysed the spatial aspects of oncogenic KRAS-related signalling activities in non-small cell lung cancer (NSCLC). Using Spa-R in conjunction with ImageJ/Fiji, we first performed annotation-guided tumour-by-tumour phenotyping using multiple signalling markers. This analysis showed histopathology-selective activation of PI3K/AKT and MAPK signalling in Kras mutant murine tumours, as well as high p38MAPK stress signalling in p53 null murine NSCLC. Subsequently, Spa-RQ was applied to measure the co-activation of MAPK, AKT, and their mutual effector mTOR pathway in individual tumours. Both murine and clinical NSCLC samples could be stratified into 'MAPK/mTOR', 'AKT/mTOR', and 'Null' signature subclasses, suggesting mutually exclusive MAPK and AKT signalling activities. Spa-RQ thus provides a robust and easy to use tool that can be employed to identify spatially-distributed tissue phenotypes

Effect of posttranslational modification on the Na+, K+ ATPase kinetics

The Na+, K+ ATPase is an essential membrane protein in eukaryotic cells, which transports Na+ out of the cell in exchange for K+ into the cell. For this transport it hydrolyses one molecule of ATP for each cycle. The partial reactions of the ATPase cycle and the effects of posttranslational modifications on ATPase activity have been studied extensively. However, amalgamation of the reported rate constants for the partial reactions along with the effect of posttranslational modifications have never been attempted. We have designed a simplified four-state mathematical model of the Na+, K+ ATPase using published results for the partial reactions. We have incorporated the effect of the Na+ allosteric site and poise dependent glutathionylation and attempted to replicate K+ activated transient currents reported in voltage clamped cardiomyocytes. Our voltage clamped cardiomyocyte results indicate the K+ activated transient is an effect of poise dependent glutathionylation rather than the Na+ subsarcolemmal space. These results can be replicated to some extent by the proposed kinetic model. This is the first kinetic model of the Na+, K+ ATPase that incorporates both partial rate constants and a reported posttranslational modification which is able to reproduce voltage clamped cardiomyocyte data

Protocol to utilize fresh uncultured human lung tumor cells for personalized functional diagnostics

Peer reviewe

Using differential reinforcement of high rates of behavior to improve work productivity : a replication and extension

Background: Due to deficits in adaptive and cognitive functioning, productivity may pose challenges for individuals with intellectual disability in the workplace.Method: Using a changing‐criterion embedded in a multiple baseline across partici‐pants design, we examined the effects of differential reinforcement of high rates of behaviour (DRH) on the rate of data entry (i.e., productivity) in four adults with intel‐lectual disability.Results: Although the DRH procedure increased the rate of correct data entry in all four participants, none of the participants achieved the criterion that we set with novice undergraduate students.Conclusions: Our results indicate that DRH is an effective intervention to increase rate of correct responding in individuals with intellectual disability, but that achiev‐ing the same productivity as workers without disability may not always be possible

Functional diagnostics using fresh uncultured lung tumor cells to guide personalized treatments

Functional profiling of a cancer patient's tumor cells holds potential to tailor personalized cancer treatment. Here, we report the utility of fresh uncultured tumor-derived EpCAM(+) epithelial cells (FUTCs) for ex vivo drug response interrogation. Analysis of murine Kras mutant FUTCs demonstrates pharmacological and adaptive signaling profiles comparable to subtype-matched cultured cells. By applying FUTC profiling on non-small cell lung cancer patient samples, we report robust drug-response data in 19 of 20 cases, with cells exhibiting targeted drug sensitivities corresponding to their oncogenic drivers. In one of these cases, an EGFR mutant lung adenocarcinoma patient refractory to osimertinib, FUTC profiling is used to guide compassionate treatment. FUTC profiling identifies selective sensitivity to disulfiram and the combination of carboplatin plus etoposide, and the patient receives substantial clinical benefit from treatment with these agents. We conclude that FUTC profiling provides a robust, rapid, and actionable assessment of personalized cancer treatment options.Peer reviewe

Protocol to utilize fresh uncultured human lung tumor cells for personalized functional diagnostics

Drug sensitivity data acquired from solid tumor-derived cultures are often unsuitable for personalized treatment guidance due to the lengthy turnaround time. Here, we present a protocol for determining ex vivo drug sensitivities using fresh uncultured human lung tumor-derived EpCAM+ epithelial cells (FUTCs). We describe steps for drug testing in FUTCs to identify tumor cell-selective single or combination therapy in 72 h of sample processing. The FUTC-based approach can also be used to predict in vivo resistance to known targeted therapies. For complete details on the use and execution of this protocol, please refer to Talwelkar et al. (2021).</p

A Smart Wireless Car Ignition System for Vehicle Security

Copyright: © 2017 Haider A, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.The paper proposes a novel car ignition system to replace the traditional wired technology and enhance vehicle security. This new system uses wireless transmissions to start the engine and hence eliminates the ignition wire behind the dashboard. It also allows the user to set a password of his/her choice to keep the system protected. A theft alarm that goes ‘’ON’’ when an unusual activity is sensed and/or when the wrong password is attempted to unlock the system is integrated in the system. Moreover, important factors such as economic feasibility, adaptability to the new vehicle technologies and customers’ preferences have been taken into consideration in the design of the proposed vehicle security system.Peer reviewedFinal Published versio

Driving pressure during general anesthesia for open abdominal surgery (DESIGNATION) : study protocol of a randomized clinical trial

Background Intraoperative driving pressure (Delta P) is associated with development of postoperative pulmonary complications (PPC). When tidal volume (V-T) is kept constant, Delta P may change according to positive end-expiratory pressure (PEEP)-induced changes in lung aeration. Delta P may decrease if PEEP leads to a recruitment of collapsed lung tissue but will increase if PEEP mainly causes pulmonary overdistension. This study tests the hypothesis that individualized high PEEP, when compared to fixed low PEEP, protects against PPC in patients undergoing open abdominal surgery. Methods The "Driving prESsure durIng GeNeral AnesThesIa for Open abdomiNal surgery trial" (DESIGNATION) is an international, multicenter, two-group, double-blind randomized clinical superiority trial. A total of 1468 patients will be randomly assigned to one of the two intraoperative ventilation strategies. Investigators screen patients aged >= 18 years and with a body mass index <= 40 kg/m(2), scheduled for open abdominal surgery and at risk for PPC. Patients either receive an intraoperative ventilation strategy with individualized high PEEP with recruitment maneuvers (RM) ("individualized high PEEP") or one in which PEEP of 5 cm H2O without RM is used ("low PEEP"). In the "individualized high PEEP" group, PEEP is set at the level at which Delta P is lowest. In both groups of the trial, V-T is kept at 8 mL/kg predicted body weight. The primary endpoint is the occurrence of PPC, recorded as a collapsed composite of adverse pulmonary events. Discussion DESIGNATION will be the first randomized clinical trial that is adequately powered to compare the effects of individualized high PEEP with RM versus fixed low PEEP without RM on the occurrence of PPC after open abdominal surgery. The results of DESIGNATION will support anesthesiologists in their decisions regarding PEEP settings during open abdominal surgery