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Pathogenic biallelic variants in PCK1 coding for the cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) cause PEPCK-C deficiency, a rare disorder of gluconeogenesis presenting with hypoglycemia, lactic acidosis, and hepatopathy. To date, there has been no systematic analysis of its phenotypic, biochemical, and genetic spectrum.All currently published individuals and a novel patient with genetically confirmed PEPCK-C deficiency were included. Clinical, biochemical, and genetic findings were analyzed. Protein and in-silico prediction score modeling was applied to analyze potential variant effects.Thirty-two individuals from 25 families were found, including one previously unreported patient. The typical biochemical pattern was hypoglycemia triggered by catabolic situations, elevated urinary concentrations of tricarboxylic acid cycle metabolites, mildly elevated alanine and aspartate aminotransferase and elevated lactate concentrations in serum. Plasma glutamine concentrations were elevated in some patients and may be a suitable marker for newborn screening. With adequate treatment, biochemical abnormalities usually normalized following a hypoglycemic episode. Symptom onset usually occurred in infancy with a broad range from neonatal age to adulthood. Regardless of the genotype, different phenotypes with a broad clinical spectrum were found. To date, eight genotypes with nine different PCK1 variants were identified, of which alleles with the recurrent variant c.925GA; p.(Gly309Arg) are predominant and appear to be endemic in the Finnish population. Protein modeling suggests altered manganese- and substrate-binding as superordinate pathomechanisms.Environmental factors appear to be the main determinant for the phenotype in patients with biallelic variants in PCK1. Based on the biochemical pattern, PEPCK-C deficiency is a recognizable cause of childhood hypoglycemia. It is a treatable disease and early diagnosis is important to prevent metabolic derailment and morbidity. Newborn screening can identify at least a sub-cohort of affected individuals through elevated glutamine concentrations in dry blood.

PDF file - 41K, QPCR validation.

PDF file - 61K, unsupervised top 10 positive and negative correlated genes - 5-FU basal.

PDF file - 62K, unsupervised top 10 positive and negative correlated genes - clinical.

PDF file - 64K, unsupervised top 10 positive and negative correlated genes - SN38 inducible.

PDF file - 32K, QPCR validations.

PDF file - 62K, unsupervised top 10 positive and negative correlated genes - 5-FU inducible.

PDF file - 63K, unsupervised top 10 positive and negative correlated genes - SN38 basal.

Chemotherapy response rates for advanced colorectal cancer remain disappointingly low, primarily because of drug resistance, so there is an urgent need to improve current treatment strategies. To identify novel determinants of resistance to the clinically relevant drugs 5-fluorouracil (5-FU) and SN38 (the active metabolite of irinotecan), transcriptional profiling experiments were carried out on pretreatment metastatic colorectal cancer biopsies and HCT116 parental and chemotherapy-resistant cell line models using a disease-specific DNA microarray. To enrich for potential chemoresistance-determining genes, an unsupervised bioinformatics approach was used, and 50 genes were selected and then functionally assessed using custom-designed short interfering RNA (siRNA) screens. In the primary siRNA screen, silencing of 21 genes sensitized HCT116 cells to either 5-FU or SN38 treatment. Three genes (RAPGEF2, PTRF, and SART1) were selected for further analysis in a panel of 5 colorectal cancer cell lines. Silencing SART1 sensitized all 5 cell lines to 5-FU treatment and 4/5 cell lines to SN38 treatment. However, silencing of RAPGEF2 or PTRF had no significant effect on 5-FU or SN38 sensitivity in the wider cell line panel. Further functional analysis of SART1 showed that its silencing induced apoptosis that was caspase-8 dependent. Furthermore, silencing of SART1 led to a downregulation of the caspase-8 inhibitor, c-FLIP, which we have previously shown is a key determinant of drug resistance in colorectal cancer. This study shows the power of systems biology approaches for identifying novel genes that regulate drug resistance and identifies SART1 as a previously unidentified regulator of c-FLIP and drug-induced activation of caspase-8. Mol Cancer Ther; 11(1); 119–31. ©2011 AACR.

In March of 1831, three American missionary families commenced service with the American Marathi Mission in Bombay. In just over three years, half of the party had died, and the survivors set sail for America. Their story is like what happened to many American missionaries at the end of the long eighteenth century who sought to expand the empire of Christ. From their voyage on the high seas to their everyday life in Bombay, they were confronted by their various states of dependence. Forced to reckon with unfamiliar social, cultural, economic, and political realities, they struggled on the margins of Anglo-Indian society. This article explores the uneasy relationship between American missionaries, who were acutely aware of their dependency on others as colonial interlopers, and the maritime empires they encountered.

The human-restricted pathogenNeisseria gonorrhoeaeascends into the upper female reproductive tract to cause damaging inflammation within the Fallopian tubes (salpingitis) and pelvic inflammatory disease (PID), increasing the risk of infertility and life-threatening ectopic pregnancy. The loss of ciliated cells from the epithelium is thought to be both a consequence of inflammation and a cause of the associated adverse sequelae. However, the links between infection, inflammation, and ciliated cell extrusion remain unresolved. With the use ofex vivocultures of human Fallopian tube paired with RNA sequencing we defined the tissue response to gonococcal challenge, identifying cytokine, chemokine, cell adhesion, and apoptosis related transcripts not previously recognized as potentiators of gonococcal PID. Unexpectedly, the cytokine IL-17C was one of the most highly induced genes. Yet, this cytokine has no previous association with gonococcal disease nor any sexually transmitted infection and thus it was selected for further characterization in our model. We show that human Fallopian tubes express the IL-17C receptor (IL-17RE) on the epithelial surface and that treatment with purified IL-17C induces pro-inflammatory cytokine secretion in addition to sloughing of the epithelium and generalized tissue damage. These results demonstrate a previously unrecognized but critical role of IL-17C in the damaging inflammation induced by gonococci in a human explant model of PID.SignificanceWe performed untargeted transcriptional analysis of the human Fallopian tube response toNeisseria gonorrhoeae. Focusing on one upregulated transcript not previously identified in any context of gonococcal infection, we show that the human Fallopian tube secretes the inflammation amplifying cytokine IL-17C in response toN. gonorrhoeaechallenge. IL-17C treatment of human Fallopian tubes stimulated pro-inflammatory cytokine secretion and was sufficient to elicit epithelial cell pathologies characteristic of pelvic inflammatory disease (sloughing, exfoliation). Thus, we identify IL-17C as a previously unrecognized component of the host response to gonococci that promotes inflammation and tissue damage within the human Fallopian tube.

Early detection of pathogens using nucleic acids in clinical samples often requires sensitivity at the single-copy level, which currently necessitates time-consuming and expensive nucleic acid amplification. Here, we describe 1) a redesigned flow cell in the shape of a trapezoid-subtracted geometric stadium, and 2) modified experimental procedures that allow for the measurement of sub-attomolar analytes in microliter quantities on a fluorescence-based waveguide biosensor. We verified our instrumental sensitivity with a 200-μL sample of a fluorescent streptavidin conjugate at 100 zM (100 zeptomolar, or 100·10−21 mol L−1) and theoretically explored the applicability of this modified sensing platform in a sandwich immunoassay format using a Langmuir adsorption model. We present assays that demonstrate specific detection of synthetic influenza A DNA (in buffer) and RNA (in saliva) oligonucleotides at the single-copy level (200 μL at 10 zM) using a fluorescent molecular beacon. Lastly, we demonstrate detection of isolated genomic influenza A RNA at a clinically relevant concentration. This work constitutes a sensitivity improvement of over twelve orders of magnitude compared to our previous nucleic acid detection work, illustrating the significant enhancements that can be gained with optimized experimental design.

Mycobacterium ulceransis the causative agent of the chronic and debilitating neglected tropical disease Buruli ulcer (BU) which mostly affects children. The early detection and treatment ofM. ulceransinfections can significantly minimize life-long disability resulting from surgical intervention. However, the disease is characterized by relatively few systemic systems as a result of complex host-pathogen interactions that have yet to be fully characterized, which has limited the development of both diagnostic and therapeutic approaches to treat BU. In this work, we study the interactions of the host immune system with two principleM. ulceransvirulence factors: mycolactone, an amphiphilic macrolide toxin, and lipoarabinomannan (LAM), a cell wall component of most mycobacterial pathogens. We observe that human lipoproteins have a profound effect on the interaction of both mycolactone and LAM with the immune system. Individually, both molecules are pro-inflammatory in the absence of serum and immunosuppressive in the presence of serum. When combined, mycolactone and LAM are immunosuppressive regardless of serum conditions. We also show that Toll-like receptor 2 (TLR2), a macrophage pathogen pattern recognition receptor, is critical for LAM immune stimulation but aids in mycolactone immunosuppression. These findings are a first step towards unraveling mycolactone-mediated immunosuppression during BU disease and may facilitate the development of effective diagnostics and therapeutics in the future.Author SummaryBuruli ulcer (BU) is a neglected tropical disease caused by the pathogenMycobacterium ulcerans. The principal virulence factors associated with it are the macrolide toxin mycolactone and the major cell wall component lipoarabinomannan (LAM). Here, we examine the impact of the amphiphilic biochemistry of mycolactone and LAM on their interaction with the human immune system. We show that both mycolactone and LAM associate with serum lipoproteins, and that this association is critical for the immune evasion seen in early-stageM. ulceransinfections. In the absence of serum, mycolactone is pro-inflammatory. Immunosuppression occurs only in the presence of human serum lipoproteins. In the presence of LAM, mycolactone is immunosuppressive, regardless of serum conditions. Immunosuppression is a hallmark of BU disease, and understanding the mechanisms of this immunosuppression can support the development of effective diagnostic and therapeutic strategies.

Near-infrared (NIR) emitting quantum dots (QDs) with emission in the biological transparency windows (NIR-I: 650–950 nm and NIR-II: 1000–1350 nm) are promising candidates for deep-tissue bioimaging...

Rapid, on-site diagnostics allow for timely intervention and response for warfighter support, environmental monitoring, and global health needs. Portable optical biosensors are being widely pursued as a means of achieving fieldable biosensing due to the potential speed and accuracy of optical detection. We recently developed the portable engineered analytic sensor with automated sampling (PEGASUS) with the goal of developing a fieldable, generalizable biosensing platform. Here, we detail the development of PEGASUS’s sensing hardware and use a test-bed system of identical sensing hardware and software to demonstrate detection of a fluorescent conjugate at 1 nM through biotin-streptavidin chemistry.

Traumatic brain injury (TBI) is not a single disease state but describes an array of conditions associated with insult or injury to the brain. While some individuals with TBI recover within a few days or months, others present with persistent symptoms that can cause disability, neuropsychological trauma, and even death. Understanding, diagnosing, and treating TBI is extremely complex for many reasons, including the variable biomechanics of head impact, differences in severity and location of injury, and individual patient characteristics. Because of these confounding factors, the development of reliable diagnostics and targeted treatments for brain injury remains elusive. We argue that the development of effective diagnostic and therapeutic strategies for TBI requires a deep understanding of human neurophysiology at the molecular level and that the framework of multiomics may provide some effective solutions for the diagnosis and treatment of this challenging condition. To this end, we present here a comprehensive review of TBI biomarker candidates from across the multiomic disciplines and compare them with known signatures associated with other neuropsychological conditions, including Alzheimer’s disease and Parkinson’s disease. We believe that this integrated view will facilitate a deeper understanding of the pathophysiology of TBI and its potential links to other neurological diseases.

Near-infrared (NIR) emitting quantum dots (QDs) with emission in the biological transparency windows (NIR-I: 650-950 nm and NIR-II: 1000-1350 nm) are promising candidates for deep-tissue bioimaging. However, they typically contain toxic heavy metals such as cadmium, mercury, arsenic, or lead. We report on the biocompatibility of high brightness CuInSe

Detection methods that do not require nucleic acid amplification are advantageous for viral diagnostics due to their rapid results. These platforms could provide information for both accurate diagnoses and pandemic surveillance. Influenza virus is prone to pandemic-inducing genetic mutations, so there is a need to apply these detection platforms to influenza diagnostics. Here, we analyzed the Fast Evaluation of Viral Emerging Risks (FEVER) pipeline on ultrasensitive detection platforms, including a waveguide-based optical biosensor and a flow cytometry bead-based assay. The pipeline was also evaluated in silico for sequence coverage in comparison to the U.S. Centers for Disease Control and Prevention’s (CDC) influenza A and B diagnostic assays. The influenza FEVER probe design had a higher tolerance for mismatched bases than the CDC’s probes, and the FEVER probes altogether had a higher detection rate for influenza isolate sequences from GenBank. When formatted for use as molecular beacons, the FEVER probes detected influenza RNA as low as 50 nM on the waveguide-based optical biosensor and 1 nM on the flow cytometer. In addition to molecular beacons, which have an inherently high background signal we also developed an exonuclease selection method that could detect 500 pM of RNA. The combination of high-coverage probes developed using the FEVER pipeline coupled with ultrasensitive optical biosensors is a promising approach for future influenza diagnostic and biosurveillance applications.

Enzyme-linked immunosorbent assays (ELISAs) are widely employed for the detection of protein targets due to their ease of use, sensitivity, and potential for high-throughput analyses. However, the use of ELISAs to detect non-protein targets such as lipids and amphiphiles is complicated by the physical properties of these molecules, which affects their association with functional surfaces and recognition ligands. Here, we developed a unique lipoprotein capture ELISA in which the natural association between lipoproteins and amphiphilic molecules facilitates detection of the target biomarker in a physiologically relevant conformation. An assay to detect the glycolipid lipoarabinomannan (LAM), a cell membrane component and virulence factor associated with Mycobacterial infections, was developed as a proof of concept.

Macrolides are a diverse class of hydrophobic compounds characterized by a macrocyclic lactone ring and distinguished by variable side chains/groups. Some of the most well characterized macrolides are toxins produced by marine bacteria, sea sponges, and other species. Many marine macrolide toxins act as biomimetic molecules to natural actin-binding proteins, affecting actin polymerization, while other toxins act on different cytoskeletal components. The disruption of natural cytoskeletal processes affects cell motility and cytokinesis, and can result in cellular death. While many macrolides are toxic in nature, others have been shown to display therapeutic properties. Indeed, some of the most well known antibiotic compounds, including erythromycin, are macrolides. In addition to antibiotic properties, macrolides have been shown to display antiviral, antiparasitic, antifungal, and immunosuppressive actions. Here, we review each functional class of macrolides for their common structures, mechanisms of action, pharmacology, and human cellular targets.

Metall-Kunststoff-Verbunde (MKV) sind eine innovative Werkstoffklasse, mit der insbesondere im Bereich des Kraft- und Schienenfahrzeugbaus eine signifikante Gewichtseinsparung erzielt werden kann. Das Scherschneiden als trennendes Fertigungsverfahren erlaubt eine hohe Ausbringungsrate bei geringen bauteilbezogenen Kosten. In diesem Beitrag wird der Einfluss der durch Scherschneiden erzeugten Schnittflächen von MKV auf eine nachfolgende Bauteilbeanspruchung vorgestellt. Metal-plastic composites (MKV) are an innovative class of materials by means of which a significant weight saving can be achieved, in particular in the area of power and rail vehicle construction. Shear cutting as a separating manufacturing process enables a high output rate with low component-related costs. This article presents the influence of cutting edges produced by shear cutting of MKV on a subsequent component stress.

Multiplex biomarker quantitation is ideal for tracking the progression of a disease, but sensitive, specific, and quantitative multiplex biosensing remains challenging. We achieved total internal reflection with two lasers in a single planar optical waveguide by coupling 532 nm laser light into the diffraction grating of a waveguide and 635 nm laser light directly into the thin film of the same waveguide. We confirmed that the evanescent fields generated in the waveguide can excite two fluorescent dyes (Alexa Fluor 532 and Alexa Fluor 647) held to the surface of the waveguide through biotin-streptavidin chemistry. This sensing concept holds significant potential as a platform for multiplex biomarker detection.

Martin Luther was and remains a controversial figure whose contentious legacy has been used to serve a variety of agendas over the centuries. Nowhere is this better seen than in the use of Luther as symbol during the Cold War years in the United States. As American Protestants responded to the social, cultural, and political changes that defined this period they re-interpreted Luther in surprising ways to suit their own needs. Drawing on film, Roman Catholic responses, debates among scholars, Pentecostal, ecumenical, and political representations, this essay argues that Luther’s memory, as a lieu de mèmoire, was used during the Cold War era to promote whatever cause or concern interpreters wanted to associate with his name and legacy.

The separation of biomarkers from blood is straightforward in most molecular biology laboratories. However, separation in resource-limited settings, allowing for the successful removal of biomarkers for diagnostic applications, is not always possible. The situation is further complicated by the need to separate hydrophobic signatures such as lipids from blood. Herein, we present a microfluidic device capable of centrifugal separation of serum from blood at the point of need with a system that is compatible with biomarkers that are both hydrophilic and hydrophobic. The cross-flow filtration device separates serum from blood as efficiently as traditional methods and retains amphiphilic biomarkers in serum for detection.

The bacterial cell envelope provides many important functions. It protects cells from harsh environments, serves as a selective permeability barrier, houses bioenergetic functions, defines sensitivity to antibacterial agents, and plays a crucial role in biofilm formation, symbiosis, and virulence. Despite the important roles of this cellular compartment, we lack a detailed understanding of the biosynthesis and remodeling of the cell envelope. Here, we report that the R. sphaeroides two-component signaling system NtrYX is a previously undescribed regulator of cell envelope processes, providing evidence that it is directly involved in controlling transcription of genes involved in cell envelope assembly, structure, and function in this and possibly other bacteria. Thus, our data report on a newly discovered process used by bacteria to assemble and remodel the cell envelope., Activity of the NtrYX two-component system has been associated with important processes in diverse bacteria, ranging from symbiosis to nitrogen and energy metabolism. In the facultative alphaproteobacterium Rhodobacter sphaeroides, loss of the two-component system NtrYX results in increased lipid production and sensitivity to some known cell envelope-active compounds. In this study, we show that NtrYX directly controls multiple properties of the cell envelope. We find that the response regulator NtrX binds upstream of cell envelope genes, including those involved in peptidoglycan biosynthesis and modification and in cell division. We show that loss of NtrYX impacts the cellular levels of peptidoglycan precursors and lipopolysaccharide and alters cell envelope structure, increasing cell length and the thickness of the periplasm. Cell envelope function is also disrupted in the absence of NtrYX, resulting in increased outer membrane permeability. Based on the properties of R. sphaeroides cells lacking NtrYX and the target genes under direct control of this two-component system, we propose that NtrYX plays a previously undescribed, and potentially conserved, role in the assembly, structure, and function of the cell envelope in a variety of bacteria.

Characterizing the structure of the Galactic interstellar medium (ISM) in three dimensions is of high importance for accurate modeling of dust emission as a foreground to the Cosmic Microwave Background (CMB). At high Galactic latitude, where the total dust content is low, accurate maps of the 3D structure of the ISM are lacking. We develop a method to quantify the complexity of the distribution of dust along the line of sight with the use of HI line emission. The method relies on a Gaussian decomposition of the HI spectra to disentangle the emission from overlapping components in velocity. We use this information to create maps of the number of clouds along the line of sight. We apply the method to: (a) the high-galactic latitude sky and (b) the region targeted by the BICEP/Keck experiment. In the North Galactic Cap we find on average three clouds per 0.2 square degree pixel while in the South the number falls to 2.5. The statistics of the number of clouds are affected by Intermediate-Velocity Clouds (IVCs), primarily in the North. The presence of IVCs results in detectable features in the dust emission measured by \textit{Planck}. We investigate the complexity of HI spectra in the BICEP/Keck region and find evidence for the existence of multiple components along the line of sight. The data (https://doi.org/10.7910/DVN/8DA5LH) and software are made publicly available, and can be used to inform CMB foreground modeling and 3D dust mapping., Comment: 30 pages (including appendices), 19 figures, ApJ in press

Nature photonics 14(6), 391 - 397 (2020). doi:10.1038/s41566-020-0607-z, The European XFEL is a hard X-ray free-electron laser (FEL) based on a high-electron-energy superconducting linear accelerator. The superconducting technology allows for the acceleration of many electron bunches within one radio-frequency pulse of the accelerating voltage and, in turn, for the generation of a large number of hard X-ray pulses. We report on the performance of the European XFEL accelerator with up to 5,000 electron bunches per second and demonstrating a full energy of 17.5 GeV. Feedback mechanisms enable stabilization of the electron beam delivery at the FEL undulator in space and time. The measured FEL gain curve at 9.3 keV is in good agreement with predictions for saturated FEL radiation. Hard X-ray lasing was achieved between 7 keV and 14 keV with pulse energies of up to 2.0 mJ. Using the high repetition rate, an FEL beam with 6 W average power was created., Published by Nature Publ. Group, London [u.a.]

The performance of sorbent-loaded hollow-fiber modules to capture carbon dioxide from the exhaust stream of a thermal power plant is analyzed. The hollow-fiber modules undergo a Rapid Temperature Swing Adsorption (RTSA) process to capture carbon dioxide. A detailed heat and mass transfer model is developed to simulate the performance of the fiber module. The thermal wave technique for heat recovery is incorporated to reduce the thermal energy consumption and reduce operating costs. A parametric study of six key operating parameters is presented, and tradeoffs in performance metrics with variations in operating parameters are discussed. The proposed process compares favorably with other adsorption-based carbon capture technologies.

We address the spatial scale, ionization structure, mass and metal content of gas at the Milky Way disk-halo interface detected as absorption in the foreground of seven closely-spaced, high-latitude halo blue horizontal branch stars (BHBs) with heights z = 3 - 14 kpc. We detect transitions that trace multiple ionization states (e.g. CaII, FeII, SiIV, CIV) with column densities that remain constant with height from the disk, indicating that the gas most likely lies within z < 3.4 kpc. The intermediate ionization state gas traced by CIV and SiIV is strongly correlated over the full range of transverse separations probed by our sightlines, indicating large, coherent structures greater than 1 kpc in size. The low ionization state material traced by CaII and FeII does not exhibit a correlation with either N$_{\rm HI}$ or transverse separation, implying cloudlets or clumpiness on scales less than 10 pc. We find that the observed ratio log(N_SiIV/ N_CIV), with a median value of -0.69+/-0.04, is sensitive to the total carbon content of the ionized gas under the assumption of either photoionization or collisional ionization. The only self-consistent solution for photoionized gas requires that Si be depleted onto dust by 0.35 dex relative to the solar Si/C ratio, similar to the level of Si depletion in DLAs and in the Milky Way ISM. The allowed range of values for the areal mass infall rate of warm, ionized gas at the disk-halo interface is 0.0003 < dM_gas / dtdA [M_sun kpc^-2 yr^-] < 0.006. Our data support a physical scenario in which the Milky Way is fed by complex, multiphase processes at its disk-halo interface that involve kpc-scale ionized envelopes or streams containing pc-scale, cool clumps., 12 figures, 4 tables, Accepted to ApJ. Revision features areal mass accretion rate calculation

The constitutively active, atypical protein kinase C, protein kinase M-ζ (PKMζ), is exclusively expressed in the brain and its expression increases following exposure to drugs of abuse. However, the limitations of currently available tools have made it difficult to examine the role of PKMζ in cocaine self-administration and relapse. The current study demonstrates that constitutive deletion of PKMζ potentiates cue-induced reinstatement of cocaine seeking and increases both food and cocaine self-administration, without affecting cue-driven food seeking in both male and female mice. Conditional deletion of PKMζ within the nucleus accumbens recapitulated the increase in cocaine taking and seeking seen in the constitutive knockout mice, but only in male animals. Site-specific knockdown of PKMζ in the nucleus accumbens had no effect on cocaine taking or seeking in female mice. Additionally, neither male nor female mice exhibited any alterations in food self-administration or cue-induced reinstatement of food seeking following accumbal deletion of PKMζ. Taken together these results indicate that PKMζ may act to dampen cocaine taking and seeking. Furthermore, these results indicate that PKMζ is playing divergent roles in reward seeking in males and females.

A new atom probe design is presented along with data showing spectral performance and selected microstructural characterization examples. The instrument includes a curved reflectron, a 532-nm laser, and an integrated, fixed-position, counter electrode in a configuration with moderate electric field enhancement that includes improvements in ease of use and cost of ownership. Both voltage-pulsed and laser-pulsed performance is shown for a variety of materials including Al, Si, W, 316 stainless steel, Inconel 718, and GaN. Characterization of grain boundaries and phase boundaries, including correlation with transmission electron backscatter diffraction results in Inconel 718, is shown. A detailed case study of the resultant microstructure between laser-beam and electron-beam additive manufacturing paths in Inconel 718 is also presented.

There is an urgent need for broad-based surveillance and diagnostic strategies that can be applied quickly and effectively to address emerging pathogens, and preferably at the point of need, as is evident with the current covid-19 crisis. Our innate immune system is an example of such a sensor – a recognition system that is capable of identifying all pathogens quickly and sensitively. Conserved pathogen associated molecular patterns (PAMPs) produced by the infectious agents are recognized by immune receptors (e.g. Toll like Receptors, TLRs) in an exquisite scheme of pattern recognition, culminating in universal early identification pipeline. Mimicking innate immune recognition in the laboratory can therefore provide a strategy for universal diagnosis of infection, and this has been the focus of our research for over a decade. There are three objectives to be addressed in order to develop a universal diagnostic platform based on innate immune recognition. They are: 1) understanding host-pathogen biology, and the development of tailored assays to address biochemically disparate pathogen associated molecular patterns in complex clinical samples, 2) development of multiplex detection assays and ultra-sensitive sensor platforms for their detection, 3) field adaptation of sensors and sample processing and 4) validation in blinded clinical samples. Herein, we present the strategy and progress in each of these areas of development, as outlined below. In bacterial pathogens, pathogen signatures recognized by our innate immune receptors are typically amphiphilic in biochemistry – lipoproteins or glycolipids. These lipidic molecules are intrinsically unstable in the aqueous milieu of blood, making them difficult to detect. Work from our team lead to the understanding that all bacterial amphiphilic pathogen associated molecular patterns associate with host lipoprotein carriers, which function as a biological taxi service for their transport in blood. Understanding this, we developed tailored sample processing pipeline and assays (membrane insertion and lipoprotein capture) for the detection of amphiphiles in serum samples. In viral pathogens, the pathogen signatures recognized by our innate immune receptors are nucleic acids – DNA and RNA. We developed a bioinformatic pipeline – FEVER- to capture the conserved aspects of these signatures and develop our recognition probes to target these pathogens, and then developed amplification-free assays for their detection using molecular beacon technology. Assays developed for both bacterial and viral signatures were applied to a novel, fieldable waveguide-based optical biosensor platform developed by our team. This platform uses single mode planar optical waveguides for bio-detection within an evanescent field, offering much greater sensitivity (10X or higher) than conventional immunoassay platforms. This fieldable sensor ( Figure: Schematic representing the fieldable universal diagnostic platform and approach. Blood is a universal sample – where all pathogens are recognized by innate immune receptors. Blood is collected and processed on site using a microfluidics sample processing cassette for biomarker extraction. The presence of the biomarker is then measured using novel tailored assays on a fieldable portable waveguide biosensor offering an answer within 15 min. Figure 1

In the mid-1930s Joy Ridderhof, a Quaker missionary, returned from her missionary work in Honduras a physically broken woman. In the process of recovering from malaria and the other illnesses that had not allowed her to remain on the mission field she began a new project that would transform how the gospel message was disseminated around the world. Ridderhof imagined the possibilities associated with proclaiming the message of Jesus through the use of phonograph records for Spanish listeners. The benefit of making sound recordings was quickly recognized by missionaries who were trying to reach largely illiterate and, in some cases, pre-literate populations. Ridderhof was soon asked to expand from her initial foray into Spanish language records to make recordings in other North American indigenous languages and, eventually, languages from around the world. This article analyses how Ridderhof managed this endeavour while embracing new media technologies to bring the sound of the gospel to the people of the world in their native tongue.