RESUMO
The development of wearable bioelectronic systems is a promising approach for optimal delivery of therapeutic treatments. These systems can provide continuous delivery of ions, charged biomolecules, and an electric field for various medical applications. However, rapid prototyping of wearable bioelectronic systems for controlled delivery of specific treatments with a scalable fabrication process is challenging. We present a wearable bioelectronic system comprised of a polydimethylsiloxane (PDMS) device cast in customizable 3D printed molds and a printed circuit board (PCB), which employs commercially available engineering components and tools throughout design and fabrication. The system, featuring solution-filled reservoirs, embedded electrodes, and hydrogel-filled capillary tubing, is assembled modularly. The PDMS and PCB both contain matching through-holes designed to hold metallic contact posts coated with silver epoxy, allowing for mechanical and electrical integration. This assembly scheme allows us to interchange subsystem components, such as various PCB designs and reservoir solutions. We present three PCB designs: a wired version and two battery-powered versions with and without onboard memory. The wired design uses an external voltage controller for device actuation. The battery-powered PCB design uses a microcontroller unit to enable pre-programmed applied voltages and deep sleep mode to prolong battery run time. Finally, the battery-powered PCB with onboard memory is developed to record delivered currents, which enables us to verify treatment dose delivered. To demonstrate the functionality of the platform, the devices are used to deliver H[Formula: see text] in vivo using mouse models and fluoxetine ex vivo using a simulated wound environment. Immunohistochemistry staining shows an improvement of 35.86% in the M1/M2 ratio of H[Formula: see text]-treated wounds compared with control wounds, indicating the potential of the platform to improve wound healing.
Assuntos
Tubo Capilar , Cicatrização , Animais , Camundongos , Dimetilpolisiloxanos , Modelos Animais de DoençasRESUMO
We report on a Computational Fluid Dynamics (CFD) study of the extra dispersion caused by the change in diameter when coupling two pieces of capillary tubing with different diameter. In this first investigation into the problem, the focus is on the typical flow rates (0.25≤F≤2µL/min) and diameters (d≤40µm) used in nano-LC, considering both the case of either a doubling or halving of the diameter. The CFD simulations allow to study the problem from a fundamental point of view, i.e., under otherwise perfect conditions (perfect alignment, zero dead-volume). Flow rates, capillary diameters, diffusion coefficients and liquid viscosities have been varied over a range relevant for nano-LC (Reynolds-numbers Re ≤ 1), with also an excursion made towards high-temperature nano-LC conditions (Re ≥ 10 and more). The extra dispersion caused by the change in diameter has been quantified via a volumetric variance σ2conn, defined in such a way that the overall dispersion across the entire capillary system can be easily reconstructed from the known analytical solutions in the individual segments. When the two capillaries are longer than their diffusion entry length, covering most of the practical cases, σ2conn converges to a limiting value σ2conn,∞ which varies to a close approximation with the square of flow rate. Under the investigated nano-LC conditions, the σ2conn,∞-values are surprisingly small (e.g., on the order of 0.01 to 0.15 nL2 in a 20 to 40µm connection) compared to the dispersion occurring in the remainder of the capillaries.
Assuntos
Tubo Capilar , Hidrodinâmica , Cromatografia Líquida/métodos , Difusão , ViscosidadeRESUMO
BACKGROUND: The COVID-19 pandemic has drastically changed the delivery of secondary care services. Self-collection of capillary blood at home can facilitate the monitoring of patients with chronic disease to support virtual clinics while mitigating the risk of SARS-CoV-2 infection and transmission. OBJECTIVE: To investigate the comparability of whole blood capillary and plasma venous samples for 15 routinely used biochemical analytes and to develop and pilot a user-friendly home-collection kit to support virtual outpatient clinical services. METHODS: To investigate the comparability of whole blood capillary and plasma venous samples for 15 routinely requested biochemical analytes, simultaneous samples of venous and capillary blood were collected in EDTA and lithium-heparin plasma separation tubes that were of 4-6 mL and 400-600 µL draw volume, respectively. Venous samples were analysed within 4 h of collection while capillary samples were kept at ambient temperature for three days until centrifugation and analysis. Analyte results that were comparable between the matrices were then piloted in a feasibility study in three outpatient clinical services. RESULTS: HbA1c, lipid profile and liver function tests were considered comparable and piloted in the patient feasibility study. The home-collect kit demonstrated good patient usability. CONCLUSION: Home collection of capillary blood could be a clinically-useful tool to deliver virtual care to patients with chronic disease.
Assuntos
Análise Química do Sangue/métodos , Coleta de Amostras Sanguíneas/métodos , COVID-19/sangue , Pandemias , SARS-CoV-2 , Adulto , Análise Química do Sangue/instrumentação , Coleta de Amostras Sanguíneas/instrumentação , Tubo Capilar , Estudos de Viabilidade , Feminino , Humanos , Londres , Masculino , Pessoa de Meia-Idade , Flebotomia/instrumentação , Flebotomia/métodos , Projetos Piloto , Consulta Remota , Autocuidado/instrumentação , Autocuidado/métodos , Inquéritos e QuestionáriosRESUMO
Liquid-filled capillary tubes are common structures in nature and engineering fields, which often function via vibration. Although liquid-solid interfacial tension plays important roles in the vibration behavior of the liquid-filled capillary tube, it remains elusive how the interfacial tension influences the natural frequency of capillary tube vibration. To address this, we developed a theory of beam-string structure to analyze the influence of liquid-solid interfacial tension on the vibration of a liquid-filled capillary cantilever. We used glass capillary tubes as a demo and experimentally validated the theory, where the reduced liquid-solid interfacial tension in a capillary tube decreases the natural frequencies of small-order modes. We then performed theoretical analysis and found that the change of elastocapillarity number, slenderness ratio and inner/outer radius ratio of capillary tubes enables: in higher order modes, a nonmonotonic change of natural frequency due to mode transformation between a beam and string; for lower order modes, decrease in the natural frequency to zero (increase from zero) due to mode disappearance (appearance). The developed theory would provide guidelines for high-accuracy design of capillary sensors.
Assuntos
Tubo Capilar , VibraçãoRESUMO
Antimicrobial resistance is recognized as one of the greatest emerging threats to public health. Antimicrobial resistant (AMR) microorganisms affect nearly 2 million people a year in the United States alone and place an estimated $20 billion burden on the healthcare system. The rise of AMR microorganisms can be attributed to a combination of overprescription of antimicrobials and a lack of accessible diagnostic methods. Delayed diagnosis is one of the primary reasons for empiric therapy, and diagnostic methods that enable rapid and accurate results are highly desirable to facilitate evidence-based treatment. This is particularly true for clinical situations at the point-of-care where access to state-of-the-art diagnostic equipment is scarce. Here, we present a capillary-based antimicrobial susceptibility testing platform (cAST), a unique approach that offers accelerated assessment of antimicrobial susceptibility in a low-cost and simple testing format. cAST delivers an expedited time-to-readout by means of optical assessment of bacteria incubated in a small capillary form factor along with a resazurin dye. cAST was designed using a combination of off-the-shelf and custom 3D-printed parts, making it extremely suitable for use in resource-limited settings. We demonstrate that growth of bacteria in cAST is approximately 25% faster than in a conventional microplate, further validate the diagnostic performance with clinical isolates, and show that cAST can deliver accurate antimicrobial susceptibility test results within 4-8 h.
Assuntos
Acinetobacter baumannii/efeitos dos fármacos , Antibacterianos/farmacologia , Enterobacter cloacae/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos dos fármacos , Tubo Capilar , Farmacorresistência Bacteriana/efeitos dos fármacos , Desenho de Equipamento , Testes de Sensibilidade Microbiana , Fenótipo , Impressão Tridimensional , Aço Inoxidável , Fatores de TempoRESUMO
INTRODUCTION: The microscopic examination of microhematocrit tubes (mHCT) has been proposed as the gold standard for acute and congenital Chagas disease diagnosis. We compared different mHCT methodologies detecting T. cruzi parasites in the blood. METHODS: The rotating method, water mount, and immersion oil methods were compared for their suitability, sensitivity, and specificity. RESULTS: The rotating method was easier, faster, and more sensitive than the others with 100% specificity. CONCLUSIONS: The rotating method is feasible for laboratory technicians with standard training in microscopic techniques and is recommended for the diagnosis of acute Chagas disease in primary health care facilities.
Assuntos
Tubo Capilar , Centrifugação/métodos , Doença de Chagas/diagnóstico , Hematócrito/métodos , Parasitemia/diagnóstico , Trypanosoma cruzi/isolamento & purificação , Animais , Doença de Chagas/sangue , Doença de Chagas/parasitologia , Serviços de Laboratório Clínico , Humanos , Parasitemia/parasitologia , Sensibilidade e EspecificidadeRESUMO
DNA hydrogels have received considerable attention in analytical science, however, some limitations still exist in the applications of intelligent hydrogels. In this paper, we describe a way to prepare gel film in a capillary tube based on the thermal reversible principle of DNA hydrogel and the principle of capillary action. Because of the slight change in the internal structure of gel, its permeability can be increased by the addition of some specific targets. The capillary behavior is thus changed due to the different permeability of the hydrogel film. The duration time of the target solution flowing through the capillary tube with a specified length is used to quantify this change. With this proposed method, ultra-trace DNA hydrogel (0.01 µL) is sufficient to realize the sensitive detection of cocaine without the aid of other instruments, which has a low detection limit (1.17 nM) and good selectivity.
Assuntos
Aptâmeros de Nucleotídeos , Tubo Capilar , Cocaína/análise , Hidrogéis , Técnicas Biossensoriais , Cocaína/urina , Desenho de Equipamento , Humanos , Limite de Detecção , PermeabilidadeRESUMO
Measuring forces from the piconewton to millinewton range is of great importance for the study of living systems from a biophysical perspective. The use of flexible micropipettes as highly sensitive force probes has become established in the biophysical community, advancing our understanding of cellular processes and microbial behavior. The micropipette force sensor (MFS) technique relies on measurement of the forces acting on a force-calibrated, hollow glass micropipette by optically detecting its deflections. The MFS technique covers a wide micro- and mesoscopic regime of detectable forces (tens of piconewtons to millinewtons) and sample sizes (micrometers to millimeters), does not require gluing of the sample to the cantilever, and allows simultaneous optical imaging of the sample throughout the experiment. Here, we provide a detailed protocol describing how to manufacture and calibrate the micropipettes, as well as how to successfully design, perform, and troubleshoot MFS experiments. We exemplify our approach using the model nematode Caenorhabditis elegans, but by following this protocol, a wide variety of living samples, ranging from single cells to multicellular aggregates and millimeter-sized organisms, can be studied in vivo, with a force resolution as low as 10 pN. A skilled (under)graduate student can master the technique in ~1-2 months. The whole protocol takes ~1-2 d to finish.
Assuntos
Caenorhabditis elegans/fisiologia , Chlamydomonas reinhardtii/fisiologia , Mecanotransdução Celular/fisiologia , Microscopia de Força Atômica/métodos , Imagem Óptica/métodos , Animais , Calibragem , Ação Capilar , Tubo Capilar , Elasticidade , Microscopia de Força Atômica/instrumentação , Microtecnologia/métodos , Imagem Óptica/instrumentação , ViscosidadeRESUMO
The preparation of a glass capillary pattered with lipid layersãon which lactate dehydrogenase (LDH) and glucose dehydrogenase (GDH) were regionally adsorbed and its application for simultaneous detection of d-glucose and l-lactate in human serum is described. A lipid layer was formed on the surface of BSA-unabsorbed octadecyltrichlorosilane (OTS) inner wall of a glass capillary. The electrostatic charge of the lipid layer was a key factor for adsorbing the enzymes on the lipid layer. The fluorescence intensities were observed at each enzyme site in the presence of diaphorase (DIA), ß-nicotinamide-adenine dinucleotide oxidized (NAD), resazurin, d-glucose and l-lactate. The fluorescence intensities at each enzyme site increased with an increase in the concentration of d-glucose and l-lactate=with the detection limits of 32 µM and 4.9 µM, respectively.
Assuntos
Técnicas Biossensoriais/métodos , Glicemia/análise , Enzimas Imobilizadas/química , Ácido Láctico/sangue , Espectrometria de Fluorescência/métodos , Técnicas Biossensoriais/instrumentação , Tubo Capilar , Vidro , Glucose/química , Glucose 1-Desidrogenase/química , Humanos , L-Lactato Desidrogenase/química , Ácido Láctico/química , Fosfolipídeos/química , Espectrometria de Fluorescência/instrumentação , Lipossomas UnilamelaresRESUMO
Abstract INTRODUCTION: The microscopic examination of microhematocrit tubes (mHCT) has been proposed as the gold standard for acute and congenital Chagas disease diagnosis. We compared different mHCT methodologies detecting T. cruzi parasites in the blood. METHODS: The rotating method, water mount, and immersion oil methods were compared for their suitability, sensitivity, and specificity. RESULTS: The rotating method was easier, faster, and more sensitive than the others with 100% specificity. CONCLUSIONS: The rotating method is feasible for laboratory technicians with standard training in microscopic techniques and is recommended for the diagnosis of acute Chagas disease in primary health care facilities.
Assuntos
Humanos , Animais , Trypanosoma cruzi/isolamento & purificação , Centrifugação/métodos , Doença de Chagas/diagnóstico , Parasitemia/diagnóstico , Tubo Capilar , Hematócrito/métodos , Sensibilidade e Especificidade , Doença de Chagas/parasitologia , Doença de Chagas/sangue , Parasitemia/parasitologia , Serviços de Laboratório ClínicoRESUMO
This protocol describes the design of capillary microfluidics for spinning bioactive (cell-laden) microfibers for three-dimensional (3D) cell culture and tissue-engineering applications. We describe the assembly of three types of microfluidic systems: (i) simple injection capillary microfluidics for the spinning of uniform microfibers; (ii) hierarchical injection capillary microfluidics for the spinning of core-shell or spindle-knot structured microfibers; and (iii) multi-barrel injection capillary microfluidics for the spinning of microfibers with multiple components. The diverse morphologies of these bioactive microfibers can be further assembled into higher-order structures that are similar to the hierarchical structures in tissues. Thus, by using different types of capillary microfluidic devices, diverse styles of microfibers with different bioactive encapsulation can be generated. These bioactive microfibers have potential applications in 3D cell culture, the mimicking of vascular structures, the creation of synthetic tissues, and so on. The whole protocol for device fabrication and microfiber spinning takes ~1 d.
Assuntos
Técnicas de Cultura de Células , Células Imobilizadas/citologia , Desenho de Equipamento , Dispositivos Lab-On-A-Chip , Engenharia Tecidual/métodos , Alginatos/química , Animais , Materiais Biocompatíveis/química , Cloreto de Cálcio/química , Tubo Capilar , Células Imobilizadas/fisiologia , Gelatina/química , Células Hep G2 , Humanos , Metacrilatos/química , Camundongos , Células NIH 3T3 , Esferoides Celulares/citologia , Esferoides Celulares/fisiologiaRESUMO
In this study, a simple, rapid, and label-free sensor was developed for detecting the enzymatic activity of catalase (CAT) with liquid crystals (LCs) confined in microcapillaries. Inside a microcapillary functionalized with n-octyltrichlorosilane, aldehyde-doped LCs anchored radially so that a pattern of straight lines was observed under a polarized optical microscope (POM). However, once hydrogen peroxide (HP) oxidized the aldehyde into carboxylic acid, which has surface activity, the orientation of the LCs at the interface changed, resulting in a distinct pattern change, from straight to crossed. In this system, the enzymatic activity of CAT could be detected as it inhibits the oxidation by decomposing HP; as a result, the pattern changed back to the straight one. From the orientational and optical shift, the enzymatic activity of CAT was detected up to a concentration of 0.8â¯fM under mild experimental conditions and 8 aM at pH 9.0. This result suggests the need for further study of microcapillary systems to develop simple and sensitive sensors for biochemical interactions.
Assuntos
Técnicas Biossensoriais/métodos , Catalase/análise , Cristais Líquidos/química , Microscopia de Polarização/métodos , Aldeídos/química , Aldeídos/metabolismo , Tubo Capilar , Peróxido de Hidrogênio/química , Peróxido de Hidrogênio/metabolismo , Ácidos Láuricos/química , Ácidos Láuricos/metabolismo , Muramidase/química , Tripsina/química , Urease/químicaRESUMO
Metabolites of new chemical entities can influence safety and efficacy of a molecule and often times need to be quantified in preclinical studies. However, synthetic standards of metabolites are very rarely available in early discovery. Alternate approaches such as biosynthesis need to be explored to generate these metabolites. Assessing the quantity and purity of these small amounts of metabolites with a nondestructive analytical procedure becomes crucial. Quantitative NMR becomes the method of choice for these samples. Recent advances in high-field NMR (>500 MHz) with the use of cryoprobe technology have helped to improve sensitivity for analysis of small microgram quantity of such samples. However, this type of NMR instrumentation is not routinely available in all laboratories. To analyze microgram quantities of metabolites on a routine basis with lower-resolution 400 MHz NMR instrument fitted with a broad band fluorine observe room temperature probe, a novel hybrid capillary tube setup was developed. To quantitate the metabolite in the sample, an artificial signal insertion for calculation of concentration observed (aSICCO) method that introduces an internally calibrated mathematical signal was used after acquiring the NMR spectrum. The linearity of aSICCO signal was established using ibuprofen as a model analyte. The limit of quantification of this procedure was 0.8 mM with 10 K scans that could be improved further with the increase in the number of scans. This procedure was used to quantify three metabolites-phenytoin from fosphenytoin, dextrophan from dextromethorphan, and 4-OH-diclofenac from diclofenac-and is suitable for minibiosynthesis of metabolites from in vitro systems.
Assuntos
Tubo Capilar , Espectroscopia de Ressonância Magnética/instrumentação , Anti-Inflamatórios não Esteroides/análise , Anti-Inflamatórios não Esteroides/farmacocinética , Calibragem , Cromatografia Líquida de Alta Pressão , Dextrorfano/análise , Ibuprofeno/análise , Ibuprofeno/farmacocinética , Espectroscopia de Ressonância Magnética/métodos , Fenitoína/análise , Padrões de Referência , Solventes , Espectrometria de Massas em Tandem , TemperaturaRESUMO
OBJECTIVE: To explore the role and mechanism of the two-kidney one-clip (2K1C)-activated Angiotensin II (Ang II) in the development of vascular damage in adjuvant-induced arthritis (AA) rats. METHODS: 2K1C rats were established in normal and AA rats for 35 days. Hypertension, endothelial dysfunction, and vascular hypertrophy induced by 2K1C-activated Ang II in systemic inflammation rats were evaluated. The levels of Ang II and TNF-α in serum were observed by ELISA kits. Expressions of Ang II/ATR/ERK1/2 signaling pathway molecules in the aorta were tested by immunohistochemistry or western blot. The migration and capillary tube formation abilities of human umbilical vein endothelial cells (HUVECs) were tested by migration chamber and capillary tube formation assays. RESULTS: The level of Ang II in serum was significantly increased in 2K1C rats. Compared with AA rats, the high level of Ang II activated by 2K1C reduced the endothelium-dependent vasodilator responses to acetylcholine (ACh) in the thoracic aorta and exacerbated endothelial dysfunction and vascular hypertrophy. Expressions of ATR, GRK2, p-ERK1/2, and p-NF-κB were significantly increased in the aorta of AA combined with 2K1C rats. The migration and capillary tube formation abilities of HUVECs were significantly enhanced by Ang II and TNF-α co-stimulations in vitro through the ATR/ERK1/2 signaling pathway compared to those stimulated with TNF-α. CONCLUSIONS: 2K1C-activated Ang II is involved in aggravated vascular injury and endothelial dysfunction through the ATR/ERK1/2 signaling pathway in AA rats.
Assuntos
Angiotensina II/metabolismo , Artrite/patologia , Proteínas Mutadas de Ataxia Telangiectasia , Hipertensão Renovascular/metabolismo , Sistema de Sinalização das MAP Quinases , Animais , Pressão Sanguínea/efeitos dos fármacos , Vasos Sanguíneos/efeitos dos fármacos , Vasos Sanguíneos/patologia , Tubo Capilar , Movimento Celular , Modelos Animais de Doenças , Células Endoteliais da Veia Umbilical Humana , Humanos , Hipertensão Renovascular/patologia , Masculino , NF-kappa B/efeitos dos fármacos , Ratos , Ratos Sprague-DawleyRESUMO
INTRODUCTION: Many secondary metabolites in plants are labile compounds which under environmental stress, are difficult to detect and track due to the lack of rapid in situ identification techniques, making plant metabolomics research difficult. Therefore, developing a reliable analytical method for rapid in situ identification of labile compounds and their short-lived intermediates in plants is of great importance. OBJECTIVE: To develop under atmospheric pressure, a rapid in situ method for effective identification of labile compounds and their short-lived intermediates in fresh plants. METHODOLOGY: An in vivo nanospray high-resolution mass spectrometry (HR-MS) method was used for rapid capture of labile compounds and their short-lived intermediates in plants. A quartz capillary was partially inserted into fresh plant tissues, and the liquid flowed out through the capillary tube owing to the capillary effect. A high direct current (d.c.) voltage was applied to the plant to generate a spray of charged droplets from the tip of the capillary carrying bioactive molecules toward the inlet of mass spectrometer for full-scan and MS/MS analysis. RESULTS: Many labile compounds and short-lived intermediates were identified via this method: including glucosinolates and their short-lived intermediates (existing for only 10 s) in Raphanus sativus roots, alliin and its conversion intermediate (existing for 20 s) in Allium sativum and labile precursor compound chlorogenic acid in Malus pumila Mill. CONCLUSION: The method is an effective approach for in situ identification of internal labile compounds and their short-lived intermediates in fresh plants and it can be used as an auxiliary tool to explore the degradation mechanisms of new labile plant compounds. Copyright © 2016 John Wiley & Sons, Ltd.
Assuntos
Ácido Clorogênico/química , Cisteína/análogos & derivados , Alho/química , Glucosinolatos/química , Malus/química , Raphanus/química , Espectrometria de Massas em Tandem , Tubo Capilar , Ácido Clorogênico/isolamento & purificação , Cisteína/química , Cisteína/isolamento & purificação , Glucosinolatos/isolamento & purificação , Metabolômica , Estrutura Molecular , Raízes de Plantas/química , Quartzo , Estresse Fisiológico , Fatores de TempoRESUMO
Biofilms are multicellular bacterial structures that adhere to surfaces and often endow the bacterial population with tolerance to antibiotics and other environmental insults. Biofilms frequently colonize the tubing of medical devices through mechanisms that are poorly understood. Here we studied the helicoidal spread of Pseudomonas putida biofilms through cylindrical conduits of varied diameters in slow laminar flow regimes. Numerical simulations of such flows reveal vortical motion at stenoses and junctions, which enhances bacterial adhesion and fosters formation of filamentous structures. Formation of long, downstream-flowing bacterial threads that stem from narrowings and connections was detected experimentally, as predicted by our model. Accumulation of bacterial biomass makes the resulting filaments undergo a helical instability. These incipient helices then coarsened until constrained by the tubing walls, and spread along the whole tube length without obstructing the flow. A three-dimensional discrete filament model supports this coarsening mechanism and yields simulations of helix dynamics in accordance with our experimental observations. These findings describe an unanticipated mechanism for bacterial spreading in tubing networks which might be involved in some hospital-acquired infections and bacterial contamination of catheters.
Assuntos
Biofilmes/crescimento & desenvolvimento , Tubo Capilar/microbiologia , Pseudomonas putida/fisiologia , Aderência Bacteriana , Constrição , Equipamentos e Provisões/microbiologia , Pseudomonas putida/químicaRESUMO
To develop in vivo monitoring strategies of neurotransmitters involved in brain chemistry is a challenging work for progress in understanding the roles that biomolecules play in pathology and physiology. Here we report a new type of gold nanoparticle-sheathed glass capillary nanoelectrode (Au/GCNE) for sensing cerebral dopamine. First, a size-controlled needle-type quartz capillary was pulled with a laser puller. Then, the capillary tip exterior was chemically functionalized with colloidal gold nanoparticles by the seed-mediated growth protocol. Through insulating the above tip with cathodic electrophoretic paint followed by heating to tune the exposed area of gold-nanoparticle-film, the Au/GCNE with tip apex radius ranging from ~8.9 to ~500 nm can be prepared. Scanning electron microscopy (SEM) and steady-state voltammetry were utilized to characterize the effective radius of nanoelectrodes. The results showed that the tip apex radius of Au/GCNE was mainly affected by the pre-pulled capillary tip, the modified AuNPs and the cathodic electrophoretic paint. By taking advantage of the modified AuNPs and the enhanced electrochemical performance of the nanoelectrode, a wide dynamic linear range from 2.0×10(-8) M to 5.6×10(-6) M with a low detection limit of 1.0×10(-8) M (S/N=3), as well as good selectivity for dopamine, were first achieved with the Nafion-modified Au/GCNE. In addition, the designed glass substrates of Au/GCNE were mechanically stronger and their sharp tips aided in membrane penetration during implantation in the in vivo experiment. As a result, the Nafion-modified Au/GCNE was successfully applied for amperometrically monitoring dopamine in the striatum of anesthetic rats.
Assuntos
Tubo Capilar , Condutometria/instrumentação , Corpo Estriado/metabolismo , Dopamina/metabolismo , Nanopartículas Metálicas/química , Microeletrodos , Animais , Dopamina/análise , Desenho de Equipamento , Análise de Falha de Equipamento , Vidro/química , Ouro/química , Masculino , Nanopartículas Metálicas/ultraestrutura , Microquímica/instrumentação , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
CONTEXT: Point-of-care glucose (POCG) testing on capillary blood specimens is central to maintaining glycemic control in patients with diabetes. Although there are known performance issues with POCG methods, especially for maintaining tight glucose control, there is little information about the accuracy of results in the critical ranges that may involve life-threatening conditions. OBJECTIVES: To evaluate the reliability of POCG measurements in critical, high (>600 mg/dL) and low (<40 mg/dL) ranges. DESIGN: One-year retrospective analysis of POCG (ACCU-CHEK glucose meter, Roche Diagnostics Corporation, Indianapolis, Indiana) results for routine patient care were obtained. The frequency and accuracy of repeat testing after critical POCG results was analyzed. A convenience sample of noncritical capillary POCG measurements retested on venous blood specimens by another point-of-care device (RAPIDPoint 405 analyzer, Siemens Medical Solutions USA, Malvern, Pennsylvania) was also evaluated. RESULTS: Critical values were observed in 2.4 per 1000 POCG tests (256 of 105,928; 0.24%), with the highest rate (76 of 2289; 3.32%) from the emergency department. Twice as many critical high values as critical low values were seen. Nearly 80% of critical POCG tests (204 of 256) were repeated within 10 minutes. Of these 204 repeat measurements, 112 (54.9%) met accuracy criteria (±15 mg/dL of low and ±20% of high initial values). Accuracy was significantly higher when retesting was performed on the same meter or by the same operator (P ≤ .05). Comparison of capillary and venous POCG testing of noncritical results showed no significant difference (P = .95), with 89.8% (125 of 139) meeting accuracy criteria. CONCLUSIONS: POCG measurements in the critical range are frequently erroneous, which is likely caused by preanalytic factors associated with sampling capillary blood. POCG testing practices should include retesting to confirm critical results.
Assuntos
Análise Química do Sangue/instrumentação , Glicemia/análise , Sistemas Automatizados de Assistência Junto ao Leito/estatística & dados numéricos , Doença Aguda , Análise Química do Sangue/normas , Análise Química do Sangue/estatística & dados numéricos , Tubo Capilar , Estado Terminal , Complicações do Diabetes/sangue , Complicações do Diabetes/diagnóstico , Diabetes Mellitus/sangue , Humanos , Hiperglicemia/sangue , Hiperglicemia/diagnóstico , Hipoglicemia/sangue , Hipoglicemia/diagnóstico , Sistemas Automatizados de Assistência Junto ao Leito/normas , Controle de Qualidade , Reprodutibilidade dos Testes , Estudos RetrospectivosRESUMO
In this report, protein-modified quartz nanopipettes were used to quantitatively study protein-protein interactions in attoliter sensing volumes. As shown by numerical simulations, the ionic current through the conical-shaped nanopipette is very sensitive to the surface charge variation near the pore mouth. With the appropriate modification of negatively charged human neuroglobin (hNgb) onto the inner surface of a nanopipette, we were able to detect concentration-dependent current change when the hNgb-modified nanopipette tip was exposed to positively charged cytochrome c (Cyt c) with a series of concentrations in the bath solution. Such current change is due to the adsorption of Cyt c to the inner surface of the nanopipette through specific interactions with hNgb. In contrast, a smaller current change with weak concentration dependence was observed when Cyt c was replaced with lysozyme, which does not specifically bind to hNgb. The equilibrium dissociation constant (KD) for the Cyt c-hNgb complex formation was derived and the value matched very well with the result from surface plasmon resonance measurement. This is the first quantitative study of protein-protein interactions by a conical-shaped nanopore based on charge sensing. Our results demonstrate that nanopipettes can potentially be used as a label-free analytical tool to quantitatively characterize protein-protein interactions.