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A lateral flow assay using DNA aptamer-based sensing for the detection of dopamine in urine is reported. Dopamine duplex aptamers (hybridized sensor with capture probe) are conjugated to 40-nm Au nanoparticles (AuNPs) with 20T linkers. The detection method is based on the dissociation of the duplex aptamer in the presence of dopamine, with the sensor part undergoing conformational changes and being released from the capture part. Hybridization between the complementary DNA in the test line and the conjugated AuNP-capture DNA produces a red band, whose intensity is related to the dopamine concentration. The minimum detectable concentration obtained by ImageJ analysis was <10 ng/mL (65.2 nM), while the visual limit of detection is estimated to be ~50 ng/mL (normal range of dopamine in urine of 52-480 ng/mL or 0.3-3.13 µM). No cross reactivity to other stress biomarkers in urine was confirmed. These results indicate that this robust and user-friendly point-of-care biosensor has significant potential for providing a cost-effective alternative for dopamine detection in urine.
Assuntos
Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais , Dopamina/urina , HumanosRESUMO
AIM: Microneedles (MNs) create micropunctures and deliver drugs or nutrients deep into skin layer. We demonstrated that MNs, coated with electrosprayed nanoparticles loaded with functional molecules, are useful for transdermal delivery. METHODS: Electrospraying was utilised to generate drug-loaded nanoparticles and to create uniform coating on MNs. Process parameters and release kinetics were evaluated in vitro. The in vivo efficacy of insulin-coated MNs was investigated using diabetic rats. RESULTS: Electrosprayed micro/nanoparticles loaded with dye or insulin were coated on MNs with particle size of 515 ± 286 and 522 ± 261 nm, respectively. Optimally coated MNs resulted in >70% transfer rate into porcine skins. Insulin-coated MNs were applied to diabetic rats resulting in reduction of blood glucose levels fluctuations, compared to subcutaneous injections. CONCLUSIONS: Electrospraying is shown to be an effective method to coat MNs with drug-loaded nanoparticles. Coated MNs provide a promising platform for cosmetic, drug and protein delivery applications.
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Diabetes Mellitus Experimental/tratamento farmacológico , Sistemas de Liberação de Medicamentos/instrumentação , Hipoglicemiantes/administração & dosagem , Insulina/administração & dosagem , Nanopartículas/química , Administração Cutânea , Animais , Hipoglicemiantes/farmacocinética , Hipoglicemiantes/uso terapêutico , Insulina/farmacocinética , Insulina/uso terapêutico , Agulhas , Ratos , Ratos Wistar , Pele/metabolismo , Absorção CutâneaRESUMO
Vitamin K antagonists such as warfarin are the most widely used class of oral anticoagulants. Due to a narrow therapeutic window, patients on warfarin require regular monitoring. Self-testing using point-of-care (POC) diagnostic devices is available, but cost makes this monitoring method beyond reach for many. The main objective of this research was to assess the clinical utility of a low-cost, paper-based lateral flow POC diagnostic device developed for anticoagulation monitoring without the need for a separate electronic reader. Custom-fabricated lateral flow assay (LFA) test strips comprised of a glass fiber sample pad, a nitrocellulose analytical membrane, a cellulose wicking pad, and a plastic backing card were assembled in a plastic cassette. Healthy volunteers and patients on warfarin therapy were recruited for this prospective study. For each participant, a whole blood sample was collected via fingerstick to determine: (1) international normalized ratio (INR) using the CoaguChek® XS coagulometer, (2) hematocrit by centrifugation, and (3) red blood cell (RBC) travel distance on the experimental LFA device after 240 s using digital image analysis. RBC travel distance measured on the LFA device using blood samples obtained from warfarin patients positively correlated with increasing INR value and the LFA device had the capability to statistically distinguish between healthy volunteer INR values and those for patients groups with INR ≥ 2.6. From these data, it is predicted that this low-cost, paper-based LFA device can have clinical utility for identifying anticoagulated patients taking vitamin K antagonists who are outside of the desired therapeutic efficacy window.
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Anticoagulantes/farmacologia , Equipamentos para Diagnóstico , Monitoramento de Medicamentos/instrumentação , Dispositivos Lab-On-A-Chip , Papel , Sistemas Automatizados de Assistência Junto ao Leito , Adulto , Idoso , Colódio/química , Custos e Análise de Custo , Equipamentos para Diagnóstico/economia , Monitoramento de Medicamentos/economia , Desenho de Equipamento , Feminino , Humanos , Dispositivos Lab-On-A-Chip/economia , Masculino , Membranas Artificiais , Fitas Reagentes/químicaRESUMO
Electrospun polyacrylonitrile (PAN) based carbon nanofibers (CNFs) have attracted intense attention due to their easy processing, high carbon yield, and robust mechanical properties. In this work, a CNF modified glassy carbon (GC) electrode that was coated with Nafion polymer was evaluated as a new electrode material for the simultaneous determination of trace levels of heavy metal ions by anodic stripping voltammetry (ASV). Pb(2+) and Cd(2+) were used as a representative system for this initial study. Well-defined stripping voltammograms were obtained when Pb(2+) and Cd(2+) were determined individually and then simultaneously in a mixture. Compared to a bare GC electrode, the CNF/Nafion modified GC (CNF/Nafion/GC) electrode improved the sensitivity for lead detection by 8-fold. The interface properties of the CNF/Nafion/GC were characterized by electrochemical impedance spectroscopy (EIS), which showed the importance of the ratio of CNF/Nafion on electrode performance. Under optimized conditions, the detection limits are 0.9 and 1.5 nM for Pb(2+) and Cd(2+), respectively.
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Biomarcadores/sangue , Análise Química do Sangue/métodos , Testes Hematológicos/métodos , Técnicas Analíticas Microfluídicas/métodos , Papel , Animais , Análise Química do Sangue/instrumentação , Testes Hematológicos/instrumentação , Humanos , Técnicas Analíticas Microfluídicas/instrumentação , Testes ImediatosRESUMO
Organic light-emitting diodes (OLED) were fabricated on flexible and transparent reconstituted cellulose obtained from wood pulp. Cellulose is naturally available, abundant, and biodegradable and offers a unique substrate alternative for the fabrication of flexible OLEDs. Transparent cellulose material was formed by dissolution of cellulose in an organic solvent (dimethyl acetamide) at elevated temperature (165 °C) in the presence of a salt (LiCl). The optical transmission of 40-µm thick transparent cellulose sheet averaged 85% over the visible spectrum. High brightness and high efficiency thin film OLEDs were fabricated on transparent cellulose films using phosphorescent Ir(ppy)3 as the emitter material. The OLEDs achieved current and luminous emission efficiencies as high as 47 cd A(-1) and 20 lm W(-1), respectively, and a maximum brightness of 10,000 cd m(-2).
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Pentacene-based organic thin-film transistors (OTFTs) were fabricated on several types of flexible substrate: commercial photo paper, ultra-smooth specialty paper and ultra-thin (100 µM) flexible glass. The transistors were fabricated entirely through dry-step processing. The transconductance and field-effect mobility of OTFTs on photo paper reached values of â¼0.52 mS m(-1) and â¼ 0.1 cm(2) V (-1) s(-1), respectively. Preliminary results on the lifetime of OTFTs on photo paper yielded stable transconductance and mobility values over a period of more than 250 h. The comparable characteristics of OTFTs fabricated on widely available, low cost paper and high quality expensive liquid crystal display glass indicate the potential importance of cellulose-based electronic devices.
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When knowledge has advanced to a state that includes a predictive understanding of the relationship between genome sequence and organism phenotype it will be possible for future engineers to design and produce synthetic organisms. However, the possibility of synthetic biology does not necessarily guarantee its feasibility, in much the same way that the possibility of a brute force attack fails to ensure the timely breaking of robust encryption. The size and range of natural genomes, from a few million base pairs for bacteria to over 100 billion base pairs for some plants, suggests it is necessary to evaluate the practical limits of designing genomes of similar complexity. This analysis characterizes the complexity of natural genomes, compares them to existing engineering benchmarks, and shows that existing large software programs are on similar scale with the genome of complex natural organisms.
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Benchmarking , Genoma Bacteriano/genética , Genoma de Planta/genética , Software , Bactérias/genética , Engenharia Genética , Fenótipo , Plantas/genética , Biologia SintéticaRESUMO
Cerebral spinal fluid (CSF) leakage is a major postoperative complication requiring surgical intervention, resulting in prolonged healing and higher costs. Biocompatible polymers, such as cyanoacrylates, are currently used as tissue adhesives for closing surgical defects and incisions. Coupling these polymers with nanofiber technology shows promising results for generating nanofibers used in wound care, tissue engineering, and drug delivery. Fiber membranes formed by electrospinning of n-octyl-2-cyanoacrylate (NOCA) are investigated for in situ dural closures after neurosurgery to improve the quality of the closure and prevent post-surgical CSF leaks. Electrospun NOCA fiber membranes showed significantly higher sealing capabilities of defects in human dura, with an average burst pressure of 149 mmHg, compared with that of an FDA-approved common dural sealant that had an average burst pressure of 37 mmHg. In this study, microfabrication of NOCA fibers demonstrates a promising technique for dural repairs.
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Adesivos Teciduais , Vazamento de Líquido Cefalorraquidiano/prevenção & controle , Vazamento de Líquido Cefalorraquidiano/cirurgia , Cianoacrilatos/farmacologia , Dura-Máter/cirurgia , Humanos , Polímeros , Adesivos Teciduais/farmacologiaRESUMO
A novel approach to control the area of anchor-free droplet interface bilayer (DIB) lipid membranes is presented. Unsupported DIB lipid membranes are formed at the interface of phospholipid-coated aqueous droplets dispensed in dodecane oil. Using electrodes inserted into the droplets, an external voltage is applied which modulates the effective DIB area. Electrical (capacitance or current) and optical (imaging of DIB lateral length) recordings were simultaneously performed. Alpha-hemolysin (αHL) single channel insertions into the DIB were recorded. Currents across the DIB were measured as a function of voltage and αHL concentration in the droplets. Nonlinear response is observed for current, DIB lateral length and area, and capacitance with respect to voltage. Voltage induced changes in interfacial tension modulated the DIB-oil contact angle and the membrane contact length, which provided control of membrane dimensions. Comparison of these results is made to the electrowetting effect, which is also governed by effect of voltage on the interfacial tension. This approach provides active control of the number of ion channels inserted into the DIB.
Assuntos
Fontes de Energia Elétrica , Bicamadas Lipídicas/química , Membranas Artificiais , Eletrodos , Tamanho da Partícula , Prata/química , Propriedades de SuperfícieRESUMO
Electrospun fibers consisting of poly(3,4-ethylenedioxythiophene)/poly(styrene sulfonate) (PEDOT/PSS) and poly(ethylene oxide) (PEO) have been used to successfully encapsulate and stabilize thylakoid membrane vesicles isolated from spinach. Light-driven electronic properties were measured. Fibers with immobilized thylakoids show higher electrical conductivity compared with fibers without thylakoids under white light conditions. This is attributed to the electron-generating photosynthetic reactions from the thylakoids. Electron and optical microscopy show the presence of thylakoid vesicles within the fibers using lipid-specific stains. After electrospinning into fibers, the thylakoid vesicles still exhibit an ability to produce a light-driven electron gradient, indicating that activity is preserved during the electrospinning process. These electrospun fibers provide an excellent example of incorporating photosynthetic function into an artificial system.
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Condutividade Elétrica , Luz , Polímeros , Tilacoides/fisiologia , Cápsulas/química , Células Imobilizadas , Transporte de Elétrons , Nanofibras , Fotossíntese , Spinacia oleracea/ultraestruturaRESUMO
We have developed a disposable point-of-care (POC) aptamer-based biosensor for the detection of salivary cortisol. Nonstressful and noninvasive sampling of saliva compared to that of blood makes saliva an attractive biological matrix in developing POC devices for biomarker monitoring. Aptamers are attractive as recognition elements for multiple reasons, including their specific chemical synthesis, high stability, lack of immunogenicity, and cell-free evolution. A duplex aptamer conjugated to the surface of Au nanoparticles (AuNPs) by Au-S bonds is utilized as the sensor probe in a lateral flow assay (LFA) device. The addition of saliva samples containing cortisol makes the cortisol-aptamer undergo conformational changes and dissociate from the capture probe. Increasing cortisol concentration in the dispensed saliva sample results in increased dissociation and leads to increased binding of AuNP conjugate on the test line. Therefore, the color intensity of the test line on the LFA is a direct function of the concentration of cortisol in saliva. This simple and fast method provides detection in the cortisol range of â¼0.5-15 ng/mL, which is in the clinically accepted range for salivary cortisol. The limit of detection was 0.37 ng/mL, and the accuracy was confirmed by enzyme-linked immunosorbent assay (ELISA) testing results. High selectivity was observed for salivary cortisol against other closely related steroids and stress biomarkers present in saliva.
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Floating gastro-retentive delivery systems can prolong the gastric residence providing sustained drug release. In this study, we report on self-inflating effervescence-based electrospun nanofiber membranes embedding polyethylene oxide/sodium bicarbonate cast films. In this system, sodium bicarbonate results in an effervescence effect by creating carbon dioxide gas upon contacting an acidic gastric fluid, with the resulting gas bubbles being entrapped within the swollen network of nanofibers. Eudragit RL and RS polymers are utilized as a host material to manipulate release kinetics of incorporated drugs. Pramipexole, a common medication for chronic Parkinson's disease (PD), is used as a model drug. Uniform and bead-free nanofibers with diameters of ~300 nm were obtained. Although floating nanofibers initially exhibited high water contact angles (WCA), water droplets were quickly absorbed into the surface and the WCA decreased to ~0° within 60 s. Floating lag time, total floating time, swelling properties and drug release profiles were investigated both in a simulated gastric fluid (pH 1.2 buffer solution) and in a simulated intestinal fluid (pH 6.8 buffer solution) at 37 °C. All floating nanofiber formulations began to float instantly with nearly zero floating lag time and did not sink into the solution even after 24 h. By comparison, the same formulations without sodium bicarbonate cast films could not maintain continuous floating beyond 15 min. The floating nanofiber pouches presented lower initial release of between 20 and 57 %, compared to that of non-floating nanofiber pouches (40-82% within 2 h). Clearly, floating nanofibers reduced the initial burst release and provided sustained drug release. This demonstrates the potential to result in 'once-a-day' oral introduction of drugs that normally must be taken frequently. Effervescence-based floating nanofibers present a novel and promising prototype delivery system for the drug delivery in the upper gastro-intestinal (GI) tract.
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Resinas Acrílicas/química , Preparações de Ação Retardada/química , Nanofibras/química , Polietilenoglicóis/química , Polímeros/química , Pramipexol/química , Bicarbonato de Sódio/química , Composição de Medicamentos/métodos , Sistemas de Liberação de Medicamentos/métodos , Liberação Controlada de Fármacos , Nanofibras/ultraestruturaRESUMO
We present a rapid and quantitative point-of-care (PoC) system based on a smartphone application that is capable of accurately tracking the flow of red blood cells (RBCs) through a no-reaction lateral flow assay (nrLFA) device. Utilizing only the camera feed from the smartphone and built-in image processing, the nrLFA is identified and RBC fluid flow distances and rates are recorded in parallel with the test without the need of any custom hardware or enclosure. We demonstrated the application by first measuring and then calculating hematocrit (Hct) values of whole blood samples with nominal content of 28%, 35%, 40%, and 45% Hct on the nrLFA platform. The PoC system was able to accurately measure (to within 1% Hct of nominal values) whole blood Hct in ~10-20 s after sample dispensing.
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Técnicas Biossensoriais , Aplicativos Móveis , Hematócrito , Sistemas Automatizados de Assistência Junto ao Leito , SmartphoneRESUMO
Highly efficient preconcentration is a crucial prerequisite to the identification of important protein biomarkers with extremely low abundance in target biofluids. In this work, poly(dimethylsiloxane) microchips integrated with 10 nm polycarbonate nanopore membranes were utilized as high-speed protein accumulators. Double-sided injection control of electrokinetic fluid flow in the sample channel resulted in highly localized protein accumulation at a very sharp point in the channel cross point. This greatly enhanced the ability to detect very low levels of initial protein concentration. Fluorescein labeled human serum albumin solutions of 30 and 300 pM accumulated to 3 and 30 microM in only 100 s. Initial solutions as low as 0.3 and 3 pM could be concentrated within 200 s to 0.3 and 3 microM, respectively. This demonstrates a approximately 10(5)-10(6) accumulation factor, and an accumulation rate as high as 5000/sec, yielding a >10x improvement over most results reported to date.
Assuntos
Eletroforese em Microchip/instrumentação , Membranas Artificiais , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Cimento de Policarboxilato/química , Proteínas/isolamento & purificação , Dimetilpolisiloxanos/química , Eletroforese em Microchip/economia , Eletroforese em Microchip/métodos , Desenho de Equipamento , Fluoresceína-5-Isotiocianato , Humanos , Técnicas Analíticas Microfluídicas/economia , Filtros Microporos , Pressão , Proteínas/análise , Albumina Sérica/análise , Albumina Sérica/isolamento & purificação , Fatores de Tempo , VácuoRESUMO
We report a novel label-free quantitative detection of human performance "stress" biomarkers in different body fluids based on optical absorbance of the biomarkers in the ultraviolet (UV) region. Stress biomarker (hormones and neurotransmitters) concentrations in bodily fluids (blood, sweat, urine, saliva) predict the physical and mental state of the individual. The stress biomarkers primarily focused on in this manuscript are cortisol, serotonin, dopamine, norepinephrine, and neuropeptide Y. UV spectroscopy of stress biomarkers performed in the 190-400 nm range has revealed primary and secondary absorption peaks at near-UV wavelengths depending on their molecular structure. UV characterization of individual and multiple biomarkers is reported in various biofluids. A microfluidic/optoelectronic platform for biomarker detection is reported, with a prime focus toward cortisol evaluation. The current limit of detection of cortisol in sweat is â¼200 ng/mL (â¼0.5 µM), which is in the normal (healthy) range. Plasma samples containing both serotonin and cortisol resulted in readily detectable absorption peaks at 203 (serotonin) and 247 (cortisol) nm, confirming feasibility of simultaneous detection of multiple biomarkers in biofluid samples. UV spectroscopy performed on various stress biomarkers shows a similar increasing absorption trend with concentration. The detection mechanism is label free, applicable to a variety of biomarker types, and able to detect multiple biomarkers simultaneously in various biofluids. A microfluidic flow cell has been fabricated on a polymer substrate to enable point-of-use/care UV measurement of target biomarkers. The overall sensor combines sample dispensing and fluid transport to the detection location with optical absorption measurements with a UV light emitting diode (LED) and photodiode. The biomarker concentration is indicated as a function of photocurrent generated at the target wavelength.
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Biomarcadores/análise , Testes de Química Clínica/instrumentação , Dispositivos Ópticos , Saliva/química , Suor/química , Absorção de Radiação , Métodos Analíticos de Preparação de Amostras , Biomarcadores/sangue , Biomarcadores/urina , Humanos , Dispositivos Lab-On-A-Chip , Raios UltravioletaRESUMO
The formation of fibers by electrospinning has experienced explosive growth in the past decade, recently reaching 4,000 publications and 1,500 patents per year. This impressive growth of interest is due to the ability to form fibers with a variety of materials, which lend themselves to a large and rapidly expanding set of applications. In particular, coaxial electrospinning, which forms fibers with multiple core-sheath layers from different materials in a single step, enables the combination of properties in a single fiber that are not found in nature in a single material. This article is a detailed review of coaxial electrospinning: basic mechanisms, early history and current status, and an in-depth discussion of various applications (biomedical, environmental, sensors, energy, catalysis, textiles). We aim to provide readers who are currently involved in certain aspects of coaxial electrospinning research an appreciation of other applications and of current results.
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Interstitial chemotherapy plays a pivotal role in the treatment of glioblastoma multiforme (GBM), an aggressive form of primary brain cancer, by enhancing drug biodistribution to the tumor and avoiding systemic toxicities. The use of new polymer structures that extend the release of cytotoxic agents may therefore increase survival and prevent recurrence. A novel core-sheath fiber loaded with the drug carmustine (BCNU) was evaluated in an in vivo brain tumor model. Three-dimensional discs were formed from coaxially electrospun fiber membranes and in vitro BCNU release kinetics were measured. In vivo survival was assessed following implantation of discs made of compressed core-sheath fibers (NanoMesh) either concurrently with or five days after intracranial implantation of 9L gliosarcoma. Co-implantation of NanoMesh and 9L gliosarcoma resulted in statistically significant long-term survival (>150 days). Empty control NanoMesh confirmed the safety of these novel implants. Similarly, Day 5 studies showed significant median, overall, and long-term survival rates, suggesting optimal control of tumor growth, confirmed with histological and immunohistochemical analyses. Local chemotherapy by means of biodegradable NanoMesh implants is a new treatment paradigm for the treatment for brain tumors. Drug delivery with coaxial core-sheath structures benefits from high drug loading, controlled long-term release kinetics, and slow polymer degradation. This represents a promising evolution for the current treatment of GBM.
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Implantes Absorvíveis , Antineoplásicos/administração & dosagem , Neoplasias Encefálicas/tratamento farmacológico , Preparações de Ação Retardada/química , Sistemas de Liberação de Medicamentos/métodos , Glioblastoma/tratamento farmacológico , Animais , Antineoplásicos/uso terapêutico , Liberação Controlada de Fármacos , Feminino , Membranas Artificiais , Polímeros/química , Ratos , Ratos Endogâmicos F344RESUMO
Bright blue-light emission at 410 nm is observed from Mg(2+)-doped GaN nanoparticles prepared by the nitridation of Ga(2)MgO(4) nanoparticles at 950 degrees C. The sintering of these nanoparticles during high-temperature nitridation was prevented by mixing the Ga(2)MgO(4) precursor nanoparticles with La(2)O(3) as an inert matrix before the nitridation process. The Mg(2+)-doped GaN nanoparticles were isolated from the matrix by etching with 10 % nitric acid. The Mg(2+)-doped GaN nanoparticles were characterized by photoluminescence, atomic force microscopy, X-ray diffraction, and IR analyses.
Assuntos
Gálio/química , Luz , Magnésio/química , Nanopartículas Metálicas/química , Temperatura , Cátions Bivalentes/química , Cor , Nanopartículas Metálicas/ultraestrutura , Microscopia de Força Atômica , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios XRESUMO
Hormones produced by glands in the endocrine system and neurotransmitters produced by the nervous system control many bodily functions. The concentrations of these molecules in the body are an indication of its state, hence the use of the term biomarker. Excess concentrations of biomarkers, such as cortisol, serotonin, epinephrine, and dopamine, are released by the body in response to a variety of conditions, for example, emotional state (euphoria, stress) and disease. The development of simple, low-cost modalities for point-of-use (PoU) measurements of biomarkers levels in various bodily fluids (blood, urine, sweat, saliva) as opposed to conventional hospital or lab settings is receiving increasing attention. This paper starts with a review of the basic properties of 12 primary stress-induced biomarkers: origin in the body (i.e., if they are produced as hormones, neurotransmitters, or both), chemical composition, molecular weight (small/medium size molecules and polymers, ranging from â¼100 Da to â¼100 kDa), and hydro- or lipophilic nature. Next is presented a detailed review of the published literature regarding the concentration of these biomarkers found in several bodily fluids that can serve as the medium for determination of the condition of the subject: blood, urine, saliva, sweat, and, to a lesser degree, interstitial tissue fluid. The concentration of various biomarkers in most fluids covers a range of 5-6 orders of magnitude, from hundreds of nanograms per milliliter (â¼1 µM) down to a few picograms per milliliter (sub-1 pM). Mechanisms and materials for point-of-use biomarker sensors are summarized, and key properties are reviewed. Next, selected methods for detecting these biomarkers are reviewed, including antibody- and aptamer-based colorimetric assays and electrochemical and optical detection. Illustrative examples from the literature are discussed for each key sensor approach. Finally, the review outlines key challenges of the field and provides a look ahead to future prospects.