Open-Path Laser Absorption Sensor for Mobile Measurements of Atmospheric Ammonia.

Anthropogenic emissions of ammonia to the atmosphere, particularly those from agricultural sources, can be damaging to the environment and human health and can drive a need for sensor technologies that can be used to detect and quantify the emissions. Mobile sensing approaches that can be deployed on ground-based or aerial vehicles can provide scalable solutions for high throughput measurements but require relatively compact and low-power sensor systems. This contribution presents an ammonia sensor based on wavelength modulation spectroscopy (WMS) integrated with a Herriott multi-pass cell and a quantum cascade laser (QCL) at 10.33 µm oriented to mobile use. An open-path configuration is used to mitigate sticky-gas effects and achieve high time-response. The final sensor package is relatively small (~20 L), lightweight (~3.5 kg), battery-powered (<30 W) and operates autonomously. Details of the WMS setup and analysis method are presented along with laboratory tests showing sensor accuracy (<~2%) and precision (~4 ppb in 1 s). Initial field deployments on both ground vehicles and a fixed-wing unmanned aerial vehicle (UAV) are also presented.

Functions of Thrombospondin-1 in the Tumor Microenvironment.

The identification of thrombospondin-1 as an angiogenesis inhibitor in 1990 prompted interest in its role in cancer biology and potential as a therapeutic target. Decreased thrombospondin-1 mRNA and protein expression are associated with progression in several cancers, while expression by nonmalignant cells in the tumor microenvironment and circulating levels in cancer patients can be elevated. THBS1 is not a tumor suppressor gene, but the regulation of its expression in malignant cells by oncogenes and tumor suppressor genes mediates some of their effects on carcinogenesis, tumor progression, and metastasis. In addition to regulating angiogenesis and perfusion of the tumor vasculature, thrombospondin-1 limits antitumor immunity by CD47-dependent regulation of innate and adaptive immune cells. Conversely, thrombospondin-1 is a component of particles released by immune cells that mediate tumor cell killing. Thrombospondin-1 differentially regulates the sensitivity of malignant and nonmalignant cells to genotoxic stress caused by radiotherapy and chemotherapy. The diverse activities of thrombospondin-1 to regulate autophagy, senescence, stem cell maintenance, extracellular vesicle function, and metabolic responses to ischemic and genotoxic stress are mediated by several cell surface receptors and by regulating the functions of several secreted proteins. This review highlights progress in understanding thrombospondin-1 functions in cancer and the challenges that remain in harnessing its therapeutic potential.

Cavity Ring-Down Methane Sensor for Small Unmanned Aerial Systems.

We present the development, integration, and testing of an open-path cavity ring-down spectroscopy (CRDS) methane sensor for deployment on small unmanned aerial systems (sUAS). The open-path configuration used here (without pump or flow-cell) enables a low mass (4 kg) and low power (12 W) instrument that can be readily integrated to sUAS, defined here as having all-up mass of <25 kg. The instrument uses a compact telecom style laser at 1651 nm (near-infrared) and a linear 2-mirror high-finesse cavity. We show test results of flying the sensor on a DJI Matrice 600 hexacopter sUAS. The high sensitivity of the CRDS method allows sensitive methane detection with a precision of ~10-30 ppb demonstrated for actual flight conditions. A controlled release setup, where known mass flows are delivered, was used to simulate point-source methane emissions. Examples of methane plume detection from flight tests suggest that isolated plumes from sources with a mass flow as low as ~0.005 g/s can be detected. The sUAS sensor should have utility for emissions monitoring and quantification from natural gas infrastructure. To the best of our knowledge, it is also the first CRDS sensor directly deployed onboard an sUAS.

Open-path cavity ring-down methane sensor for mobile monitoring of natural gas emissions.

We present the design, development, and testing results of a novel laser-based cavity ring-down spectroscopy (CRDS) sensor for methane detection. The sensor is specifically oriented for mobile (i.e. vehicle deployed) monitoring of natural gas emissions from oil and infrastructure. In contrast to most commercial CRDS sensors, we employ an open-path design which allows higher temporal response and a lower power and mass package more suited to vehicle integration. The system operates in the near-infrared (NIR) at 1651 nm with primarily telecom components and includes cellular communication for wireless data transfer. Along with basic sensor design and lab testing, we present results of field measurements showing performance over a range of ambient conditions and examples of methane plume detection.

CD47 Receptor Globally Regulates Metabolic Pathways That Control Resistance to Ionizing Radiation.

Modulating tissue responses to stress is an important therapeutic objective. Oxidative and genotoxic stresses caused by ionizing radiation are detrimental to healthy tissues but beneficial for treatment of cancer. CD47 is a signaling receptor for thrombospondin-1 and an attractive therapeutic target because blocking CD47 signaling protects normal tissues while sensitizing tumors to ionizing radiation. Here we utilized a metabolomic approach to define molecular mechanisms underlying this radioprotective activity. CD47-deficient cells and cd47-null mice exhibited global advantages in preserving metabolite levels after irradiation. Metabolic pathways required for controlling oxidative stress and mediating DNA repair were enhanced. Some cellular energetics pathways differed basally in CD47-deficient cells, and the global declines in the glycolytic and tricarboxylic acid cycle metabolites characteristic of normal cell and tissue responses to irradiation were prevented in the absence of CD47. Thus, CD47 mediates signaling from the extracellular matrix that coordinately regulates basal metabolism and cytoprotective responses to radiation injury.

Hydrogen sulfide is an endogenous potentiator of T cell activation.

H(2)S is an endogenous signaling molecule that may act via protein sulfhydrylation to regulate various physiological functions. H(2)S is also a byproduct of dietary sulfate metabolism by gut bacteria. Inflammatory bowel diseases such as ulcerative colitis are associated with an increase in the colonization of the intestine by sulfate reducing bacteria along with an increase in H(2)S production. Consistent with its increased production, H(2)S is implicated as a mediator of ulcerative colitis both in its genesis or maintenance. As T cells are well established mediators of inflammatory bowel disease, we investigated the effect of H(2)S exposure on T cell activation. Using primary mouse T lymphocytes (CD3+), OT-II CD4+ T cells, and the human Jurkat T cell line, we show that physiological levels of H(2)S potentiate TCR-induced activation. Nanomolar levels of H(2)S (50-500 nM) enhance T cell activation assessed by CD69 expression, interleukin-2 expression, and CD25 levels. Exposure of T cells to H(2)S dose-dependently enhances TCR-stimulated proliferation with a maximum at 300 nM (30% increase, p < 0.01). Furthermore, activation increases the capacity of T cells to make H(2)S via increased expression of cystathionine γ-lyase and cystathionine ß-synthase. Disrupting this response by silencing these H(2)S producing enzymes impairs T cell activation, and proliferation and can be rescued by the addition of 300 nM H(2)S. Thus, H(2)S represents a novel autocrine immunomodulatory molecule in T cells.

A homogeneous SIRPα-CD47 cell-based, ligand-binding assay: Utility for small molecule drug development in immuno-oncology.

CD47 is an immune checkpoint protein that downregulates both the innate and adaptive anti-tumor immune response via its counter receptor SIRPα. Biologics, including humanized CD47 monoclonal antibodies and decoy SIRPα receptors, that block the SIRPα-CD47 interaction, are currently being developed as cancer immunotherapy agents. However, adverse side effects and limited penetration of tumor tissue associated with their structure and large size may impede their clinical application. We recently developed a quantitative high throughput screening assay platform to identify small molecules that disrupt the binding of SIRPα and CD47 as an alternative approach to these protein-based therapeutics. Here, we report on the development and optimization of a cell-based binding assay to validate active small molecules from our biochemical screening effort. This assay has a low volume, high capacity homogenous format that relies on laser scanning cytometry (LSC) and associated techniques to enhance signal to noise measurement of cell surface binding. The LSC assay is specific, concentration dependent, and validated for the two major human SIRPα variants (V1 and V2), with results that parallel those of our biochemical data as well as published studies. We also utilized the LSC assay to confirm published studies showing that the inhibition of amino-terminal pyroglutamate formation on CD47 using the glutaminyl cyclase inhibitor SEN177 disrupts SIRPα binding. The SIRPα-CD47 interaction could be quantitatively measured in live and fixed tumor cells. Use of fixed cells reduces the burden of cell maintenance and provides stable cell standards to control for inter- and intra-assay variations. We also demonstrate the utility of the assay to characterize the activity of the first reported small molecule antagonists of the SIRPα-CD47 interaction. This assay will support the screening of thousands of compounds to identify or validate active small molecules as hits, develop structure activity relationships and assist in the optimization of hits to leads by a typical iterative medicinal chemistry campaign.

Fr-PPIChem: An Academic Compound Library Dedicated to Protein-Protein Interactions.

Protein-protein interactions (PPIs) mediate nearly every cellular process and represent attractive targets for modulating disease states but are challenging to target with small molecules. Despite this, several PPI inhibitors (iPPIs) have entered clinical trials, and a growing number of PPIs have become validated drug targets. However, high-throughput screening efforts still endure low hit rates mainly because of the use of unsuitable screening libraries. Here, we describe the collective effort of a French consortium to build, select, and store in plates a unique chemical library dedicated to the inhibition of PPIs. Using two independent predictive models and two updated databases of experimentally confirmed PPI inhibitors developed by members of the consortium, we built models based on different training sets, molecular descriptors, and machine learning methods. Independent statistical models were used to select putative PPI inhibitors from large commercial compound collections showing great complementarity. Medicinal chemistry filters were applied to remove undesirable structures from this set (such as PAINS, frequent hitters, and toxic compounds) and to improve drug likeness. The remaining compounds were subjected to a clustering procedure to reduce the final size of the library while maintaining its chemical diversity. In practice, the library showed a 46-fold activity rate enhancement when compared to a non-iPPI-enriched diversity library in high-throughput screening against the CD47-SIRPα PPI. The Fr-PPIChem library is plated in 384-well plates and will be distributed on demand to the scientific community as a powerful tool for discovering new chemical probes and early hits for the development of potential therapeutic drugs.

Quantitative high-throughput screening assays for the discovery and development of SIRPα-CD47 interaction inhibitors.

CD47 is an immune checkpoint molecule that downregulates key aspects of both the innate and adaptive anti-tumor immune response via its counter receptor SIRPα, and it is expressed at high levels in a wide variety of tumor types. This has led to the development of biologics that inhibit SIRPα engagement including humanized CD47 antibodies and a soluble SIRPα decoy receptor that are currently undergoing clinical trials. Unfortunately, toxicological issues, including anemia related to on-target mechanisms, are barriers to their clinical advancement. Another potential issue with large biologics that bind CD47 is perturbation of CD47 signaling through its high-affinity interaction with the matricellular protein thrombospondin-1 (TSP1). One approach to avoid these shortcomings is to identify and develop small molecule molecular probes and pretherapeutic agents that would (1) selectively target SIRPα or TSP1 interactions with CD47, (2) provide a route to optimize pharmacokinetics, reduce on-target toxicity and maximize tissue penetration, and (3) allow more flexible routes of administration. As the first step toward this goal, we report the development of an automated quantitative high-throughput screening (qHTS) assay platform capable of screening large diverse drug-like chemical libraries to discover novel small molecules that inhibit CD47-SIRPα interaction. Using time-resolved Förster resonance energy transfer (TR-FRET) and bead-based luminescent oxygen channeling assay formats (AlphaScreen), we developed biochemical assays, optimized their performance, and individually tested them in small-molecule library screening. Based on performance and low false positive rate, the LANCE TR-FRET assay was employed in a ~90,000 compound library qHTS, while the AlphaScreen oxygen channeling assay served as a cross-validation orthogonal assay for follow-up characterization. With this multi-assay strategy, we successfully eliminated compounds that interfered with the assays and identified five compounds that inhibit the CD47-SIRPα interaction; these compounds will be further characterized and later disclosed. Importantly, our results validate the large library qHTS for antagonists of CD47-SIRPα interaction and suggest broad applicability of this approach to screen chemical libraries for other protein-protein interaction modulators.

Teleconferencing model for forensic consultation, court testimony, and continuing education.

A medical center-based forensic clinic that provides the necessary comprehensive consultation, continuing education, court testimony, and clinical services through an applied model of teleconferencing applications is addressed. Telemedicine technology and services have gained the attention of both legal and clinical practitioners, examining trends and models of health care for underserved populations, and identifying where consultation with a team of professionals may benefit service providers in rural communities. The contribution offered herein provides an understanding of the history of the development of the clinic, a theoretical model that has been applied to a clinical forensic program that employs telepsychiatry services, and the ethical and malpractice liability issues confronted in using teleconferencing services. This model is examined through a child and adolescent forensic evaluation clinic. The goals of this model are offered, as are a number of applications within the broad spectrum of services utilizing telemedicine. Finally, changing patterns are addressed in clinically based health-care delivery for criminal justice, social services, and forensic mental health.

Data obtained with a novel approach to estimate installment loan acquisition costs.

The data presented in this study were obtained from a novel approach to estimate a comprehensive loan acquisition cost. The latter includes commute costs and wage losses in addition to the monthly installment payments. These cost estimates represent the monetary value (in U.S. dollars) of the costs of driving to and from the installment lender storefront and that of the potential hourly wage losses, that is, wage loss from the driving time and the time spent at the loan office filling out the required paperwork to obtain the loan. Borrowers only get the net loan proceeds, that is, the original loan amount minus the comprehensive loan acquisition costs. The study area has 160 counties. It was created from the ESRI ArcGIS Map (a mapping software) using the spatial data from the U.S. Census, Topologically Integrated Geographic Encoding and Referencing (TIGER) Cartographic boundary files representing the geographies of states and counties. Using the U.S. road networks, the origin of the trip is a county seat in Arkansas and the destination of the trip is a county seat in a surrounding state of Tennessee, Mississippi, Louisiana, Texas, Oklahoma, and Missouri. The transportation networks were established using Google Earth/Directions to efficiently measure the travel time (distance). The average cost of a trip of 17 cents (U.S. dollar) was calculated based on the U.S. Department of Transportation Survey data, which identify important attributes of a typical vehicle used in a county such as model make, age of the vehicle, fuel consumption, etc. There are 10 occupational industry sectors where a typical borrower has a job. To estimate wage loss, the data were gathered from the U.S. Department of Labor, Bureau of Labor Statistics, namely, the Occupational Employment Statistics. Putting the missing pieces together, the data contain in this study improve our understanding of extra costs borne by borrowers located in the "loan desert" area. As expected, interior counties post high loan acquisition costs compared with border counties. The data from this study are useful to the public, businesses, policymakers, and researchers working on consumer finance.

Trauma, change, and psychological health in the 21st century.

The impact of stressful life events on health has been the object of inquiry for decades. Health care professionals have studied how stressful life events may precipitate or contribute to the onset of illness. Traumatic events and experiences can profoundly affect physical and psychological well-being, which in turn may predispose an individual to greater resilience or greater vulnerability to life stresses. Examined herein is the relationship between life stresses--including social stressors, political stressors, and environmental stressors--and the critical health related issues that psychologists need to prepare for in both the science and the practice of psychology over the next decade and during the 21st century.

Allied health professionals with 2020 vision.

Allied health professionals in all disciplines must be visionary as they address education, training, and health care delivery in the next decade. Examined herein are forces of change in education, training, health care, the recognition of essential leadership styles, and the paradigm shifts facing the allied health profession in the health care arena. Some visionary directions are offered for allied health professionals to consider as health policy and clinical agendas emerge toward the year 2020.

Biological activity of designed photolabile metal nitrosyls: light-dependent activation of soluble guanylate cyclase and vasorelaxant properties in rat aorta.

The biological and pharmacological utility of nitric oxide (NO) has led to the development of many classes of NO-donor compounds as both research tools and therapeutic agents. Many donors currently in use rely on thermal decomposition or bioactivation for the release of NO. We have developed several photolabile metal-nitrosyl donors that release NO when exposed to either visible or UV light. Herein, we show that these donors are capable of activating the primary "NO receptor", soluble guanylate cyclase (sGC), in a light-dependent fashion leading to increases in cGMP. Moreover, we demonstrate that these donors are capable of eliciting light-dependent increases of cGMP in smooth muscle cells and vasorelaxation of rat aortic smooth muscle tissue, all effects that are attributed to activation of sGC. The potential utility of these compounds as drugs and/or research tools is discussed.

Reducing alcohol-impaired driving crashes through the use of social marketing.

Over the past decade there has been little decrease in the number of alcohol-related driving fatalities. During this time most interventions have been educational or legal. This paper presents the results of a field experiment that used social marketing to introduce a new ride program into three rural communities. Almost all people in the 21-34-year-old target know that they should not drive while impaired, and most agree it is not a good thing to do, but for many the opportunity to behave properly does not exist. The Road Crew program was developed using new product development techniques and implemented by developing broad coalitions within the communities. A key feature of the program included rides to, between, and home from bars in older luxury vehicles. Results showed a significant shift in riding/driving behavior, especially among 21-34-year olds, a projected 17% decline in alcohol-related crashes in the first year, no increase in drinking behavior, and large savings between the reactive cost of cleaning up after a crash and the proactive cost of avoiding a crash. Programs have become self-sustaining based on fares and tavern contributions, and have become part of the life style in the treatment communities.

Secreted Thrombospondin-1 Regulates Macrophage Interleukin-1ß Production and Activation through CD47.

Thrombospondin-1 regulates inflammation by engaging several cell surface receptors and by modulating activities of other secreted factors. We have uncovered a novel role of thrombospondin-1 in modulating production and activation of the proinflammatory cytokine IL-1ß by human and murine macrophages. Physiological concentrations of thrombospondin-1 limit the induction by lipopolysaccharide of IL-1ß mRNA and total protein production by human macrophages. This inhibition can be explained by the ability of thrombospondin-1 to disrupt the interaction between CD47 and CD14, thereby limiting activation of NFκB/AP-1 by lipopolysaccharide. Only the CD47-binding domain of thrombospondin-1 exhibits this activity. In contrast, CD47, CD36, and integrin-binding domains of thrombospondin-1 independently enhance the inflammasome-dependent maturation of IL-1ß in human THP-1 monocyte-derived macrophages. Correspondingly, mouse bone marrow-derived macrophages that lack either thrombospondin-1 or CD47 exhibit diminished induction of mature IL-1ß in response to lipopolysaccharide. Lack of CD47 also limits lipopolysaccharide induction of IL-1ß, NLRP3, and caspase-1 mRNAs. These data demonstrate that thrombospondin-1 exerts CD47-dependent and -independent pro-and anti-inflammatory effects on the IL-1ß pathway. Therefore, thrombospondin-1 and its receptor CD47 may be useful targets for limiting the pro-inflammatory effects of lipopolysaccharide and for treating endotoxemia.

Telepsychiatry: critical dimensions for forensic services.

The use of telepsychiatry technology and services has gained attention among legal and clinical practitioners. In the current article, telepsychiatry is defined, and an innovative model of telepsychiatry care delivery that is in use in a child and adolescent forensic evaluation clinic is examined. Critical factors specific to forensics services are examined, as are those specific to telepsychiatry, including transmission mode, privacy and confidentiality, expense, quality of care, face-to-face versus video transmission, user satisfaction, and liability concerns in the use of telepsychiatry.

CD47-dependent regulation of H2S biosynthesis and signaling in T cells.

Pharmacological concentrations of H2S donors inhibit some T cell functions by inhibiting mitochondrial function, but evidence is also emerging that H2S at physiological concentrations produced via chemical sources and endogenously is a positive physiological mediator of T cell function. Expression of the H2S biosynthetic enzymes cystathionine γ-lyase (CSE) and cystathionine ß-synthase (CBS) is induced in response to T cell receptor signaling. Inhibiting the induction of these enzymes limits T cell activation and proliferation, which can be overcome by exposure to exogenous H2S at submicromolar concentrations. Exogenous H2S at physiological concentrations increases the ability of T cells to form an immunological synapse by altering cytoskeletal actin dynamics and increasing the reorientation of the microtubule-organizing center. Downstream, H2S enhances T cell receptor-dependent induction of CD69, CD25, and Interleukin-2 (IL-2) gene expression. The T cell stimulatory activity of H2S is enhanced under hypoxic conditions that limit its oxidative metabolism by mitochondrial and nonenzymatic processes. Studies of the receptor CD47 have revealed the first endogenous inhibitory signaling pathway that regulates H2S signaling in T cells. Binding of the secreted protein thrombospondin-1 to CD47 elicits signals that block the stimulatory activity of exogenous H2S on T cell activation and limit the induction of CSE and CBS gene expression. CD47 signaling thereby inhibits T cell receptor-mediated T cell activation.