Identification of Novel Series of Potent and Selective Relaxin Family Peptide Receptor 1 (RXFP1) Agonists.

Relaxin H2 is a clinically relevant peptide agonist for relaxin family peptide receptor 1 (RXFP1), but a combination of this hormone's short plasma half-life and the need for injectable delivery limits its therapeutic potential. We sought to overcome these limitations through the development of a potent small molecule (SM) RXFP1 agonist. Although two large SM HTS campaigns failed in identifying suitable hit series, we uncovered novel chemical space starting from the only known SM RXFP1 agonist series, represented by ML290. Following a design-make-test-analyze strategy based on improving early dose to man ranking, we discovered compound 42 (AZ7976), a highly selective RXFP1 agonist with sub-nanomolar potency. We used AZ7976, its 10 000-fold less potent enantiomer 43 and recombinant relaxin H2 to evaluate in vivo pharmacology and demonstrate that AZ7976-mediated heart rate increase in rats was a result of RXFP1 agonism. As a result, AZ7976 was selected as lead for continued optimization.

Discovery of Clinical Candidate AZD5462, a Selective Oral Allosteric RXFP1 Agonist for Treatment of Heart Failure.

Optimization of the highly potent and selective, yet metabolically unstable and poorly soluble hRXFP1 agonist AZ7976 led to the identification of the clinical candidate, AZD5462. Assessment of RXFP1-dependent cell signaling demonstrated that AZD5462 activates a highly similar panel of downstream pathways as relaxin H2 but does not modulate relaxin H2-mediated cAMP second messenger responsiveness. The therapeutic potential of AZD5462 was assessed in a translatable cynomolgus monkey heart failure model. Following 8 weeks of treatment with AZD5462, robust improvements in functional cardiac parameters including LVEF were observed at weeks 9, 13, and 17 without changes in heart rate or mean arterial blood pressure. AZD5462 was well tolerated in both rat and cynomolgus monkey and has successfully completed phase I studies in healthy volunteers. In summary, AZD5462 is a small molecule pharmacological mimetic of relaxin H2 signaling at RXFP1 and holds promise as a potential therapeutic approach to treat heart failure patients.

Microwave and Antenna Systems in Medical Applications.

The non-ionizing nature of microwave radiation and the low cost of microwave electronics offer innovative solutions for medical diagnosis, treatment, and health monitoring [...].

A wearable microwave instrument can detect and monitor traumatic abdominal injuries in a porcine model.

Abdominal injury is a frequent cause of death for trauma patients, and early recognition is essential to limit fatalities. There is a need for a wearable sensor system for prehospital settings that can detect and monitor bleeding in the abdomen (hemoperitoneum). This study evaluates the potential for microwave technology to fill that gap. A simple prototype of a wearable microwave sensor was constructed using eight antennas. A realistic porcine model of hemoperitoneum was developed using anesthetized pigs. Ten animals were measured at healthy state and at two sizes of bleeding. Statistical tests and a machine learning method were used to evaluate blood detection sensitivity. All subjects presented similar changes due to accumulation of blood, which dampened the microwave signal ([Formula: see text]). The machine learning analysis yielded an area under the receiver operating characteristic (ROC) curve (AUC) of 0.93, showing 100% sensitivity at 90% specificity. Large inter-individual variability of the healthy state signal complicated differentiation of bleedings from healthy state. A wearable microwave instrument has potential for accurate detection and monitoring of hemoperitoneum, with automated analysis making the instrument easy-to-use. Future hardware development is necessary to suppress measurement system variability and enable detection of smaller bleedings.

Discrete Dipole Approximation-Based Microwave Tomography for Fast Breast Cancer Imaging.

This paper describes a fast microwave tomography reconstruction algorithm based on the two-dimensional discrete dipole approximation. Synthetic data from a finite-element based solver and experimental data from a microwave imaging system are used to reconstruct images and to validate the algorithm. The microwave measurement system consists of 16 monopole antennas immersed in a tank filled with lossy coupling liquid and a vector network analyzer. The low-profile antennas and lossy nature of system make the discrete dipole approximation an ideal forward solver in the image reconstructions. The results show that the algorithm can readily reconstruct a 2D plane of a cylindrical phantom. The proposed forward solver combined with the nodal adjoint method for computing the Jacobian matrix enables the algorithm to reconstruct an image within 6 seconds. This implementation provides a significant time savings and reduced memory requirements and is a dramatic improvement over previous implementations.

Nephrotoxic antisense oligonucleotide SPC5001 induces kidney injury biomarkers in a proximal tubule-on-a-chip.

Antisense oligonucleotides (ASOs) are a promising therapeutic modality. However, failure to predict acute kidney injury induced by SPC5001 ASO observed in a clinical trial suggests the need for additional preclinical models to complement the preceding animal toxicity studies. To explore the utility of in vitro systems in this space, we evaluated the induction of nephrotoxicity and kidney injury biomarkers by SPC5001 in human renal proximal tubule epithelial cells (HRPTEC), cultured in 2D, and in a recently developed kidney proximal tubule-on-a-chip. 2D HRPTEC cultures were exposed to the nephrotoxic ASO SPC5001 or the safe control ASO 556089 (0.16-40 µM) for up to 72 h, targeting PCSK9 and MALAT1, respectively. Both ASOs induced a concentration-dependent downregulation of their respective mRNA targets but cytotoxicity (determined by LDH activity) was not observed at any concentration. Next, chip-cultured HRPTEC were exposed to SPC5001 (0.5 and 5 µM) and 556089 (1 and 10 µM) for 48 h to confirm downregulation of their respective target transcripts, with 74.1 ± 5.2% for SPC5001 (5 µM) and 79.4 ± 0.8% for 556089 (10 µM). During extended exposure for up to 20 consecutive days, only SPC5001 induced cytotoxicity (at the higher concentration; 5 µM), as evaluated by LDH in the perfusate medium. Moreover, perfusate levels of biomarkers KIM-1, NGAL, clusterin, osteopontin and VEGF increased 2.5 ± 0.2-fold, 3.9 ± 0.9-fold, 2.3 ± 0.6-fold, 3.9 ± 1.7-fold and 1.9 ± 0.4-fold respectively, in response to SPC5001, generating distinct time-dependent profiles. In conclusion, target downregulation, cytotoxicity and kidney injury biomarkers were induced by the clinically nephrotoxic ASO SPC5001, demonstrating the translational potential of this kidney on-a-chip.

Expansion of the Nodal-Adjoint Method for Simple and Efficient Computation of the 2D Tomographic Imaging Jacobian Matrix.

This paper focuses on the construction of the Jacobian matrix required in tomographic reconstruction algorithms. In microwave tomography, computing the forward solutions during the iterative reconstruction process impacts the accuracy and computational efficiency. Towards this end, we have applied the discrete dipole approximation for the forward solutions with significant time savings. However, while we have discovered that the imaging problem configuration can dramatically impact the computation time required for the forward solver, it can be equally beneficial in constructing the Jacobian matrix calculated in iterative image reconstruction algorithms. Key to this implementation, we propose to use the same simulation grid for both the forward and imaging domain discretizations for the discrete dipole approximation solutions and report in detail the theoretical aspects for this localization. In this way, the computational cost of the nodal adjoint method decreases by several orders of magnitude. Our investigations show that this expansion is a significant enhancement compared to previous implementations and results in a rapid calculation of the Jacobian matrix with a high level of accuracy. The discrete dipole approximation and the newly efficient Jacobian matrices are effectively implemented to produce quantitative images of the simplified breast phantom from the microwave imaging system.

A Multicompartment Human Kidney Proximal Tubule-on-a-Chip Replicates Cell Polarization-Dependent Cisplatin Toxicity.

Drug-induced kidney injury is a major clinical problem and causes drug attrition in the pharmaceutical industry. To better predict drug-induced kidney injury, kidney in vitro cultures with enhanced physiologic relevance are developed. To mimic the proximal tubule, the main site of adverse drug reactions in the kidney, human-derived renal proximal tubule epithelial cells (HRPTECs) were injected in one of the channels of dual-channel Nortis chips and perfused for 7 days. Tubes of HRPTECs demonstrated expression of tight junction protein 1 (zona occludens-1), lotus lectin, and primary cilia with localization at the apical membrane, indicating an intact proximal tubule brush border. Gene expression of cisplatin efflux transporters multidrug and toxin extrusion transporter (MATE) 1 (SLC47A1) and MATE2-k (SLC47A2) and megalin endocytosis receptor increased 19.9 ± 5.0-, 23.2 ± 8.4-, and 106 ± 33-fold, respectively, in chip cultures compared with 2-dimensional cultures. Moreover, organic cation transporter 2 (OCT2) (SLC22A2) was localized exclusively on the basolateral membrane. When infused from the basolateral compartment, cisplatin (25 µM, 72 hours) induced toxicity, which was evident as reduced cell number and reduced barrier integrity compared with vehicle-treated chip cultures. Coexposure with the OCT2 inhibitor cimetidine (1 mM) abolished cisplatin toxicity. In contrast, infusion of cisplatin from the apical compartment did not induce toxicity, which was in line with polarized localization of cisplatin uptake transport proteins, including OCT2. In conclusion, we developed a dual channel human kidney proximal tubule-on-a-chip with a polarized epithelium, restricting cisplatin sensitivity to the basolateral membrane and suggesting improved physiologic relevance over single-compartment models. Its implementation in drug discovery holds promise to improve future in vitro drug-induced kidney injury studies. SIGNIFICANCE STATEMENT: Human-derived kidney proximal tubule cells retained characteristics of epithelial polarization in vitro when cultured in the kidney-on-a-chip, and the dual-channel construction allowed for drug exposure using the physiologically relevant compartment. Therefore, cell polarization-dependent cisplatin toxicity could be replicated for the first time in a kidney proximal tubule-on-a-chip. The use of this physiologically relevant model in drug discovery has potential to aid identification of safe novel drugs and contribute to reducing attrition rates due to drug-induced kidney injury.

Investigation of an Ultra Wideband Noise Sensor for Health Monitoring.

Quick on-scene assessment and early intervention is the key to reduce the mortality of stroke and trauma patients, and it is highly desirable to develop ambulance-based diagnostic and monitoring devices in order to provide additional support to the medical personnel. We developed a compact and low cost ultra wideband noise sensor for medical diagnostics and vital sign monitoring in pre-hospital settings. In this work, we demonstrated the functionality of the sensor for respiration and heartbeat monitoring. In the test, metronome was used to manipulate the breathing pattern and the heartbeat rate reference was obtained with a commercial electrocardiogram (ECG) device. With seventeen tests performed for respiration rate detection, sixteen of them were successfully detected. The results also show that it is possible to detect the heartbeat rate accurately with the developed sensor.

Hyperthermia Treatment Planning Including Convective Flow in Cerebrospinal Fluid for Brain Tumour Hyperthermia Treatment Using a Novel Dedicated Paediatric Brain Applicator.

Hyperthermia therapy (40-44 °C) is a promising option to increase efficacy of radiotherapy/chemotherapy for brain tumours, in particular paediatric brain tumours. The Chalmers Hyperthermia Helmet is developed for this purpose. Hyperthermia treatment planning is required for treatment optimisation, but current planning systems do not involve a physically correct model of cerebrospinal fluid (CSF). This study investigates the necessity of fluid modelling for treatment planning. We made treatments plans using the Helmet for both pre-operative and post-operative cases, comparing temperature distributions predicted with three CSF models: a convective "fluid" model, a non-convective "solid" CSF model, and CSF models with increased effective thermal conductivity ("high-k"). Treatment plans were evaluated by T90, T50 and T10 target temperatures and treatment-limiting hot spots. Adequate heating is possible with the helmet. In the pre-operative case, treatment plan quality was comparable for all three models. In the post-operative case, the high-k models were more accurate than the solid model. Predictions to within ±1 °C were obtained by a 10-20-fold increased effective thermal conductivity. Accurate modelling of the temperature in CSF requires fluid dynamics, but modelling CSF as a solid with enhanced effective thermal conductivity might be a practical alternative for a convective fluid model for many applications.

Airborne contamination and surgical site infection: Could a thirty-year-old idea help solve the problem?

Surgical site infection (SSI) is a most serious postoperative complication, associated with increased morbidity and mortality, as well as extended therapy and elevated healthcare costs. In open clean surgery, e.g. orthopedic and cardiothoracic operations, the risk of SSI is strongly correlated with the amount of airborne bacteria being present in the operating room and the surgical field. The source of these bacteria is the surgical team itself, as we all emit thousands of bacteria-carrying skin particles every minute into the air. Although the risk of airborne contamination has decreased over the years, thanks to modern surgical clothing and advanced operating room ventilation, airborne bacteria are still detected and cause SSI. However, during the past thirty years there has been a simple and potentially effective preventive method waiting to be noticed. In 1986 Hall, Mackintosh and Hoffman found in a controlled experimental study that the application of regular unperfumed skin lotion to a person's body reduced the emission of airborne bacteria-carrying particles by approximately 90%. Moreover, the effect lasted at least 4 hours, which corresponds to a major surgical procedure. Thus, in the light of those results the present paper puts forth the hypothesis that this method can decrease the incidence of airborne bacterial contamination and SSI in open clean surgery. The paper also discusses the rationale and advantages of the method, and questions why it has escaped scientific attention for so long. In healthcare, difficult problems rarely have a simple and cheap solution. However, the use of ordinary skin lotion in open surgery may just be one, as it could potentially help prevent surgical site infection, and thereby increase patient safety and reduce healthcare costs.

3D Simulations of Intracerebral Hemorrhage Detection Using Broadband Microwave Technology.

Early, preferably prehospital, detection of intracranial bleeding after trauma or stroke would dramatically improve the acute care of these large patient groups. In this paper, we use simulated microwave transmission data to investigate the performance of a machine learning classification algorithm based on subspace distances for the detection of intracranial bleeding. A computational model, consisting of realistic human head models of patients with bleeding, as well as healthy subjects, was inserted in an antenna array model. The Finite-Difference Time-Domain (FDTD) method was then used to generate simulated transmission coefficients between all possible combinations of antenna pairs. These transmission data were used both to train and evaluate the performance of the classification algorithm and to investigate its ability to distinguish patients with versus without intracranial bleeding. We studied how classification results were affected by the number of healthy subjects and patients used to train the algorithm, and in particular, we were interested in investigating how many samples were needed in the training dataset to obtain classification results better than chance. Our results indicated that at least 200 subjects, i.e., 100 each of the healthy subjects and bleeding patients, were needed to obtain classification results consistently better than chance (p < 0.05 using Student's t-test). The results also showed that classification results improved with the number of subjects in the training data. With a sample size that approached 1000 subjects, classifications results characterized as area under the receiver operating curve (AUC) approached 1.0, indicating very high sensitivity and specificity.

Application of Two-Dimensional Discrete Dipole Approximation in Simulating Electric Field of a Microwave Breast Imaging System.

The two-dimensional electric field distribution of the microwave imaging system is numerically simulated for a simplified breast tumour model. The proposed two-dimensional discrete dipole approximation (DDA) has the potential to improve computational speed compared to other numerical methods while retaining comparable accuracy. We have modeled the field distributions in COMSOL Multiphysics as baseline results to benchmark the DDA simulations. We have also investigated the adequate sampling size and the effect of inclusion size and property contrast on solution accuracy. In this way, we can utilize the 2D DDA as an alternative, fast and reliable forward solver for microwave tomography. From a mathematical perspective, the derivation of the 2D DDA and its application to microwave imaging is new and not previously implemented. The simulation results and the measurements show that the 2D DDA is a well-grounded forward solver for the specified microwave breast imaging system.

A Discrete Dipole Approximation Solver Based on the COCG-FFT Algorithm and Its Application to Microwave Breast Imaging.

We introduce the discrete dipole approximation (DDA) for efficiently calculating the two-dimensional electric field distribution for our microwave tomographic breast imaging system. For iterative inverse problems such as microwave tomography, the forward field computation is the time limiting step. In this paper, the two-dimensional algorithm is derived and formulated such that the iterative conjugate orthogonal conjugate gradient (COCG) method can be used for efficiently solving the forward problem. We have also optimized the matrix-vector multiplication step by formulating the problem such that the nondiagonal portion of the matrix used to compute the dipole moments is block-Toeplitz. The computation costs for multiplying the block matrices times a vector can be dramatically accelerated by expanding each Toeplitz matrix to a circulant matrix for which the convolution theorem is applied for fast computation utilizing the fast Fourier transform (FFT). The results demonstrate that this formulation is accurate and efficient. In this work, the computation times for the direct solvers, the iterative solver (COCG), and the iterative solver using the fast Fourier transform (COCG-FFT) are compared with the best performance achieved using the iterative solver (COCG-FFT) in C++. Utilizing this formulation provides a computationally efficient building block for developing a low cost and fast breast imaging system to serve under-resourced populations.

Effects of the Plastic of the Realistic GeePS-L2S-Breast Phantom.

A breast phantom developed at the Supelec Institute was interrogated to study its suitability for microwave tomography measurements. A microwave measurement system based on 16 monopole antennas and a vector network analyzer was used to study how the S-parameters are influenced by insertion of the phantom. The phantom is a 3D-printed structure consisting of plastic shells that can be filled with tissue mimicking liquids. The phantom was filled with different liquids and tested with the measurement system to determine whether the plastic has any effects on the recovered images or not. Measurements of the phantom when it is filled with the same liquid as the surrounding coupling medium are of particular interest. In this case, the phantom plastic has a substantial effects on the measurements which ultimately detracts from the desired images.

A novel multi-parametric high content screening assay in ciPTEC-OAT1 to predict drug-induced nephrotoxicity during drug discovery.

Drug-induced nephrotoxicity is a major concern in the clinic and hampers the use of available treatments as well as the development of innovative medicines. It is typically discovered late during drug development, which reflects a lack of in vitro nephrotoxicity assays available that can be employed readily in early drug discovery, to identify and hence steer away from the risk. Here, we report the development of a high content screening assay in ciPTEC-OAT1, a proximal tubular cell line that expresses several relevant renal transporters, using five fluorescent dyes to quantify cell health parameters. We used a validation set of 62 drugs, tested across a relevant concentration range compared to their exposure in humans, to develop a model that integrates multi-parametric data and drug exposure information, which identified most proximal tubular toxic drugs tested (sensitivity 75%) without any false positives (specificity 100%). Due to the relatively high throughput (straight-forward assay protocol, 96-well format, cost-effective) the assay is compatible with the needs in the early drug discovery setting to enable identification, quantification and subsequent mitigation of the risk for nephrotoxicity.

Test systems in drug discovery for hazard identification and risk assessment of human drug-induced liver injury.

INTRODUCTION: The liver is an important target for drug-induced toxicities. Early detection of hepatotoxic drugs requires use of well-characterized test systems, yet current knowledge, gaps and limitations of tests employed remains an important issue for drug development. Areas Covered: The current state of the science, understanding and application of test systems in use for the detection of drug-induced cytotoxicity, mitochondrial toxicity, cholestasis and inflammation is summarized. The test systems highlighted herein cover mostly in vitro and some in vivo models and endpoint measurements used in the assessment of small molecule toxic liabilities. Opportunities for research efforts in areas necessitating the development of specific tests and improved mechanistic understanding are highlighted. Expert Opinion: Use of in vitro test systems for safety optimization will remain a core activity in drug discovery. Substantial inroads have been made with a number of assays established for human Drug-induced Liver Injury. There nevertheless remain significant gaps with a need for improved in vitro tools and novel tests to address specific mechanisms of human Drug-Induced Liver Injury. Progress in these areas will necessitate not only models fit for application, but also mechanistic understanding of how chemical insult on the liver occurs in order to identify translational and quantifiable readouts for decision-making.

Clinical Evaluation of a Microwave-Based Device for Detection of Traumatic Intracranial Hemorrhage.

Traumatic brain injury (TBI) is the leading cause of death and disability among young persons. A key to improve outcome for patients with TBI is to reduce the time from injury to definitive care by achieving high triage accuracy. Microwave technology (MWT) allows for a portable device to be used in the pre-hospital setting for detection of intracranial hematomas at the scene of injury, thereby enhancing early triage and allowing for more adequate early care. MWT has previously been evaluated for medical applications including the ability to differentiate between hemorrhagic and ischemic stroke. The purpose of this study was to test whether MWT in conjunction with a diagnostic mathematical algorithm could be used as a medical screening tool to differentiate patients with traumatic intracranial hematomas, chronic subdural hematomas (cSDH), from a healthy control (HC) group. Twenty patients with cSDH and 20 HC were measured with a MWT device. The accuracy of the diagnostic algorithm was assessed using a leave-one-out analysis. At 100% sensitivity, the specificity was 75%-i.e., all hematomas were detected at the cost of 25% false positives (patients who would be overtriaged). Considering the need for methods to identify patients with intracranial hematomas in the pre-hospital setting, MWT shows promise as a tool to improve triage accuracy. Further studies are under way to evaluate MWT in patients with other intracranial hemorrhages.

Compact self-grounded Bow-Tie antenna design for an UWB phased-array hyperthermia applicator.

Using UWB hyperthermia systems has the potential to improve the heat delivery to deep seated tumours. In this paper, we present a novel self-grounded Bow-Tie antenna design which is to serve as the basis element in a phased-array applicator. The UWB operation in the frequency range of 0.43-1 GHz is achieved by immersing the antenna in a water bolus. The radiation characteristics are improved by appropriate shaping the water bolus and by inclusion of dielectric layers on the top of the radiating arms of the antenna. In order to find the most appropriate design, we use a combination of performance indicators representing the most important attributes of the antenna. These are the UWB impedance matching, the transmission capability and the effective field size. The antenna was constructed and experimentally validated on muscle-like phantom. The measured reflection and transmission coefficients as well as radiation characteristics are in excellent agreement with the simulated results. MR image acquisitions with antenna located inside MR bore indicate a negligible distortion of the images by the antenna itself, which indicates MR compatibility.

Microwave technology for detecting traumatic intracranial bleedings: tests on phantom of subdural hematoma and numerical simulations.

Traumatic brain injury is the leading cause of death and severe disability for young people and a major public health problem for elderly. Many patients with intracranial bleeding are treated too late, because they initially show no symptoms of severe injury and are not transported to a trauma center. There is a need for a method to detect intracranial bleedings in the prehospital setting. In this study, we investigate whether broadband microwave technology (MWT) in conjunction with a diagnostic algorithm can detect subdural hematoma (SDH). A human cranium phantom and numerical simulations of SDH are used. Four phantoms with SDH 0, 40, 70 and 110 mL are measured with a MWT instrument. The simulated dataset consists of 1500 observations. Classification accuracy is assessed using fivefold cross-validation, and a validation dataset never used for training. The total accuracy is 100 and 82-96 % for phantom measurements and simulated data, respectively. Sensitivity and specificity for bleeding detection were 100 and 96 %, respectively, for the simulated data. SDH of different sizes is differentiated. The classifier requires training dataset size in order of 150 observations per class to achieve high accuracy. We conclude that the results indicate that MWT can detect and estimate the size of SDH. This is promising for developing MWT to be used for prehospital diagnosis of intracranial bleedings.