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In this study, we developed a wearable nanozyme-enzyme electrochemical biosensor that enablies sweat lactate monitoring. The biosensor comprises a flexible electrode system prepared on a polyimide (PI) film and the Janus textile for unidirectional sweat transport. We obtained favorable electrochemical activities for hydrogen peroxide reduction by modifying the laser-scribed graphene (LSG) electrode with cerium dioxide (CeO2)-molybdenum disulphide (MoS2) nanozyme and gold nanoparticles (AuNPs). By further immobilisation of lactate oxidase (LOx), the proposed biosensor achieves chronoamperometric lactate detection in artificial sweat within a range of 0.1-50.0 mM, a high sensitivity of 25.58 µA mM-1cm-2 and a limit of detection (LoD) down to 0.135 mM, which fully meets the requirements of clinical diagnostics. We demonstrated accurate lactate measurements in spiked artificial sweat, which is consistent with standard ELISA results. To monitor the sweat produced by volunteers while exercising, we conducted on-body tests, showcasing the wearable biosensor's ability to provide clinical sweat lactate diagnosis for medical treatment and sports management.
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Técnicas Biossensoriais , Técnicas Eletroquímicas , Ouro , Grafite , Ácido Láctico , Oxigenases de Função Mista , Suor , Dispositivos Eletrônicos Vestíveis , Técnicas Biossensoriais/instrumentação , Suor/química , Humanos , Ácido Láctico/análise , Técnicas Eletroquímicas/instrumentação , Oxigenases de Função Mista/química , Oxigenases de Função Mista/metabolismo , Ouro/química , Grafite/química , Enzimas Imobilizadas/química , Molibdênio/química , Nanopartículas Metálicas/química , Eletrodos , Dissulfetos/química , Limite de DetecçãoRESUMO
Improving the mechanical properties of wound dressings and achieving personalized automatic real-time in situ deposition are important for accelerating wound management and repair. In this study, we report a self-designed automatic in situ deposition device based on solution blow spinning (SBS) to prepare poly(lactic-co-glycolic acid) (PLGA) and poly-L-lactic acid (PLLA) composite (PLGA/PLLA) nanofibrous membranes for wound dressing at a short distance. Polymer solution and in situ deposition conditions, including air pressure, spinning distance, solvent extrusion rate, and spinning rate, were optimized using orthogonal experiments and characterized via dynamic mechanical analysis. The microscopic morphology and physical properties of the prepared PLGA/PLLA composite nanofibrous membranes show that their strength, adhesion, water vapor transmission rate (WVTR), water retention, water absorption, degradation, and other properties were sufficient for wound-dressing applications. To investigate the possibility of a biomedical wound-dressing material, tannic acid (TA) was incorporated into the PLGA/PLLA composite nanofibrous membranes. The resultant PLGA/PLLA/TA composite nanofibrous membranes exhibited good biocompatibility and exceptional antibacterial properties against both Escherichia coli and Staphylococcus aureus. A pilot animal study illustrated the potential of this in situ deposition of PLGA/PLLA/TA composite nanofibrous membranes across multiple applications in wound healing/repair by reducing wound scar tissue formation and fibroblast overactivation.
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Antibacterianos , Bandagens , Escherichia coli , Nanofibras , Poliésteres , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Staphylococcus aureus , Cicatrização , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Nanofibras/química , Staphylococcus aureus/efeitos dos fármacos , Animais , Escherichia coli/efeitos dos fármacos , Poliésteres/química , Antibacterianos/química , Antibacterianos/farmacologia , Cicatrização/efeitos dos fármacos , Membranas Artificiais , Camundongos , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , HumanosRESUMO
BACKGROUND: Prosthetic-joint infection (PJI) is one of the severest complications after arthroplasty. However, antibiotics are not effective in the bacteria in biofilm outside the prosthetic-joint. Antimicrobial peptides have an efficient antimicrobial activity in staphylococcus aureus compared with conventional antibiotics. METHODS: Bone marrow stem cells (BMSCs) were isolated, cultured and transfected with cathelicidins antimicrobial peptides proline-arginine-rich 39 amino acid peptide (PR-39) lentivirus. The expression of PR-39 gene in BMSCs was detected by RT-PCR, and the antibacterial activity of PR-39 was measured by agar diffusion method. The transfection efficiency was detected by fluorescence microscopy. The infection model of artificial knee joint in rabbits were established. Kirschner wire was used as the knee joint implant to implant the distal femur through the femoral intercondylar fossa of rabbits. 24 rabbits were randomly divided into 2 groups for the above operations: group A was inoculated 0.5 mL into the joint cavity immediately after the incision was sutured 1 × 107 Staphylococcus aureus of colony forming unit (CFU), group B was inoculated with Staphylococcus aureus and PR-39. After operation, the wound conditions and histological changes were observed by X-ray and optical microscope respectively, CRP and erythrocyte sedimentation rate were measured by test assay. RESULTS: The transfection efficiency of lentivirus vectortransfected BMSCs was 74.09%. The supernatant of lentivirus vector had obvious inhibitory effect on Staphylococcus aureus, and the antibacterial rate was 98.43%. 100% infection observed in group A while few infection observed in group B; serum CRP and ESR at a high level in group A while decreased in group B after operation. There were no significant difference in CRP and ESR between the pLV/PR-39 group and pLV/EGFP group at day 1 and 3 respectively after surgery. However, CRP and ESR in the pLV/PR-39 groupwere significantly lower than the pLV/EGFP group at day 7 and 14 respectively after operation. CONCLUSIONS: Rabbits planted BMSCs expressing PR-39 were significantly increased resistance to Staphylococcus aureus in PJI than control group thus showing great potential for preventing implant-associated infection. It will provide a potential new therapeutic agent for implant-associated infection.
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Infecções Relacionadas à Prótese , Infecções Estafilocócicas , Animais , Coelhos , Catelicidinas , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Peptídeos Catiônicos Antimicrobianos , Infecções Estafilocócicas/prevenção & controle , Staphylococcus aureus , Infecções Relacionadas à Prótese/prevenção & controleRESUMO
Analysis of cortisol levels in human sweat is increasingly important as it can be a "stress biomarker" in stress-related disorders, giving real-time information about human health status. In this study, a portable 3D microfluidic origami biosensor based on a smartphone was developed for cortisol-level detection in human sweat. Molybdenum disulfide (MoS2) nanosheet-mediated fluorescence resonance energy transfer (FRET) and fluorescently labeled aptamers were employed in the biosensing process. A multilayer-structured 3D origami microfluidic chip was fabricated and functionalized to facilitate low-volume perspired human sweat collection, transportation, and detection. The translatability of the biosensor was exhibited by the fluorescence analysis in a smartphone mounted in a custom-designed holder. The critical design parameters of the microfluidic origami biosensor, including the characterization of various paper substrates, the concentration of MoS2 nanosheets, and the incubation/reaction time, were adjusted to obtain an acceptable range for the assay dynamic range and limit of detection (LOD). Under optimum conditions, various doses of cortisol within the physiologically relevant range of 10-1000 ng/mL reported in human sweat were tested to evaluate the performance of the proposed biosensor. It displayed an LOD of 6.76 ng/mL at 3σ in artificial sweat, an analysis time of 25 min, and high selectivity. The performance of the proposed cortisol sensor was compared with an enzyme-linked immunosorbent assay (ELISA) for a spiked artificial sweat sample, and a correlation coefficient of 0.988 was found. The proposed biosensor also presented satisfactory results in the determination of the cortisol levels in a real human sweat sample. The resulting portable biosensor provides a rapid, low-cost, convenient, and non-invasive sensing solution for the point-of-care analysis of cortisol levels in sweat.
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Técnicas Biossensoriais , Hidrocortisona , Técnicas Biossensoriais/métodos , Humanos , Hidrocortisona/análise , Limite de Detecção , Microfluídica , Suor/químicaRESUMO
In this study, an origami microfluidic electrochemical nano-aptasensor was developed for the rapid detection of the peanut allergen Ara h1. Specifically, the microfluidic aptasensor was fabricated through sequential folding of a piece of chromatography paper substrate patterned with microchannel and screen-printed electrodes. Aptamer-decorated black phosphorus nanosheets (BPNSs) were electrodeposited onto the paper-based electrode surface as sensing probes for enhanced electrochemical detection and high specificity and selectivity. Critical design parameters (the concentration of probe, time for self-assembly of aptamer and reaction time) were investigated to optimize the aptasensor performance. The prepared aptasensor was able to complete detection within 20 min and demonstrated a linear range from 50 ~ 1000 ng/mL with a detection limit of 21.6 ng/mL. The aptasensor was successfully used to detect the Ara h1 spiked cookie dough sample. The proposed method reduces the gap between complex lab testing and food allergen analysis at the point of need.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Alérgenos , Arachis , Técnicas Eletroquímicas , Eletrodos , Ouro , Limite de Detecção , MicrofluídicaRESUMO
Vibrio parahaemolyticus is one of the foodborne bacteria that widely present in seafood as well as the leading cause of seafood-associated bacterial gastroenteritis. Traditional identification of such pathogens mainly relies on culturing methods, ELISA or PCR. These methods are usually laborious, time-consuming with poor diagnosis competences, or require costly and bulky equipment though of high sensitivity. In this study, a thread-based microfluidic electrochemical aptasensor was designed, fabricated and tested by using label-free aptamer immunosensing technology for rapid and highly sensitive detection of Vibrio parahaemolyticus in seafood. Both the microfluidic channels and electrodes were simply fabricated on threads. Molybdenum disulfide (MoS2) nanosheets were used to obtain enhanced sensitivity of the electrochemical measurement. When used in detecting Vibrio parahaemolyticus, the proposed aptasensor has a dynamic detection range of 10-106 CFU mL-1 with a detection limit of 5.74 CFU mL-1. Compared with traditional plate counting method, the proposed aptasensor has higher detection sensitivity and less assay time (30 min), while high specificity and accuracy are kept. The proposed microfluidic thread-based electrochemical aptasensor grabs the potential to detect other pathogens by simply functionalizing the threaded electrodes with aptamers for targeted biological substances.
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Técnicas Biossensoriais , Vibrio parahaemolyticus , Eletrodos , Limite de Detecção , Microfluídica , Reação em Cadeia da Polimerase , Vibrio parahaemolyticus/genéticaRESUMO
In virtue of the inherent molecular recognition and programmability, DNA has recently become the most promising for high-performance biosensors. The rationally engineered nucleic acid architecture will be very advantageous to hybridization efficiency, specificity, and sensitivity. Herein, a robust and split-mode photoelectrochemical (PEC) biosensor for miRNA-196a was developed based on an entropy-driven tetrahedral DNA (EDTD) amplifier coupled with superparamagnetic nanostructures. The DNA tetrahedron structure features in rigidity and structural stability that contribute to obtain precise identification units and specific orientations, improving the hybridization efficiency, sensitivity, and selectivity of the as-designed PEC biosensor. Further, superparamagnetic Fe3O4@SiO2@CdS particles integrated with DNA nanostructures are beneficial for the construction of a split-mode, highly selective, and reliable PEC biosensor. Particularly, the enzyme- and hairpin-free EDTD amplifier eliminates unnecessary interference from the complex secondary structure of pseudoknots or kissing loops in typical hairpin DNAs, significantly lowers the background noise, and improves the detection sensitivity. This PEC biosensor is capable of monitoring miRNA-196a in practical settings with additional advantages of efficient electrode fabrication, stability, and reproducibility. This strategy can be extended to various miRNA assays in complex biological systems with excellent performance.
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Técnicas Biossensoriais , Nanoestruturas , DNA/genética , Técnicas Eletroquímicas , Entropia , Limite de Detecção , Reprodutibilidade dos Testes , Dióxido de SilícioRESUMO
Organic dyes are typically applied as photosensitizers in photoelectrochemical (PEC) cells but have not been reported in polarity-reversal-mode PEC sensors with excellent sensitivity and accuracy. Herein, an elegant and robust PEC biosensor for carcinoembryonic antigen (CEA) has been designed by photocurrent polarity switching of CdTe quantum dots (QDs), which is obtained by embedding methylene blue (MB) into amplified double-stranded DNA (dsDNA) anchored to the superparamagnetic Fe3O4@SiO2. The target-triggered Exo III-assisted cyclic amplification strategy and in situ magnetic enrichment enable the remarkable sensitivity. The extraction of target-analogue single-stranded DNA (output DNA) contributes to high selectivity resulting from the elimination of possible interferences in real samples or matrixes. Particularly, this exclusive polarity-reversal-mode PEC aptasensing can efficiently eliminate the false-positive or false-negative signals, leading to accurate measurements. Moreover, different from the probes and layer-by-layer assembled photoelectric beacons on electrodes in advance, this rational split-type approach is doomed to help the PEC biosensor with additional merits of convenient fabrication, short time consumption, wider linearity, as well as outstanding reproducibility and stability in practical applications. In light of the ability of MB acting as a kind of signal probe in typical electrochemical sensors, certainly, this ingenious design can not only be extended to a wide variety of target monitoring but also provide new ideas for the construction of high-performance electrochemical and PEC biosensors.
Assuntos
Técnicas Biossensoriais , Antígeno Carcinoembrionário/análise , DNA/química , Técnicas Eletroquímicas , Azul de Metileno/química , Compostos de Cádmio/química , Compostos Férricos/química , Humanos , Fenômenos Magnéticos , Tamanho da Partícula , Processos Fotoquímicos , Pontos Quânticos/química , Dióxido de Silício/química , Propriedades de Superfície , Telúrio/químicaRESUMO
MicroRNA (miRNA) has become a key indicator of cancer diagnosis based on its abnormal expression levels. However, high-performance monitoring of miRNA is still a difficult task because of its low concentration, small size, and similarity of sequences. Herein, an elegant and robust photoelectrochemical (PEC) biosensor for miRNA-122 has been flexibly designed based on the split mode between entropy-driven DNA signal amplification and photocurrent expression. The entropy-driven DNA circuit uses a multichain composite structure instead of a DNA hairpin structure, leading to decrease the reversibility of each step of the signal amplification system. Also, the unique increasing entropy mechanism, rather than the free energy release from the new base pairs forming, improves the reaction efficiency and enhances more thermal stability and strong specific identification ability. Particularly, the biologically functionalized superparamagnetic Fe3O4@SiO2 complex endows this split mode PEC biosensor with excellent specificity and enhanced efficiency of electrode fabrication. Additionally, this strategy of only the CdTe-signal DNA modified on the ITO electrode for photocurrent readout overcomes the shortcomings of tediously long layer-by-layer assembly process and multiple rinsing steps, leading to efficient improvement of the stability and reproducibility for the as-designed PEC biosensor. This elegant strategy opens a new path for miRNA measurements with superior performance.
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Técnicas Biossensoriais/métodos , DNA/química , Entropia , Óxido Ferroso-Férrico/química , Nanoestruturas/química , Processos Fotoquímicos , Eletroquímica , Dióxido de Silício/químicaRESUMO
Over the past decades, researchers have been seeking attractive substrate materials to keep microfluidics improving to outbalance the drawbacks and issues. Cellulose substrates, including thread, paper and hydrogels are alternatives due to their distinct structural and mechanical properties for a number of applications. Thread have gained considerable attention and become promising powerful tool due to its advantages over paper-based systems thus finds numerous applications in the development of diagnostic systems, smart bandages and tissue engineering. To the best of our knowledge, no comprehensive review articles on the topic of thread-based microfluidics have been published and it is of significance for many scientific communities working on Microfluidics, Biosensors and Lab-on-Chip. This review gives an overview of the advances of thread-based microfluidic diagnostic devices in a variety of applications. It begins with an overall introduction of the fabrication followed by an in-depth review on the detection techniques in such devices and various applications with respect to effort and performance to date. A few perspective directions of thread-based microfluidics in its development are also discussed. Thread-based microfluidics are still at an early development stage and further improvements in terms of fabrication, analytical strategies, and function to become low-cost, low-volume and easy-to-use point-of-care (POC) diagnostic devices that can be adapted or commercialized for real world applications.
Assuntos
Técnicas Biossensoriais/instrumentação , Celulose/química , Fibra de Algodão/análise , Técnicas Analíticas Microfluídicas/instrumentação , Animais , Técnicas Biossensoriais/economia , Técnicas Biossensoriais/métodos , Colorimetria/economia , Colorimetria/instrumentação , Colorimetria/métodos , Técnicas Eletroquímicas/economia , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Gossypium/química , Humanos , Técnicas Analíticas Microfluídicas/economia , Técnicas Analíticas Microfluídicas/métodos , Papel , Sistemas Automatizados de Assistência Junto ao Leito/economiaRESUMO
Electrohydrodynamics is commonly used in microfluidics to control and manipulate the fluid. Though there are studies on the rotation flow in suspended films, the thin film liquid is easily broken and cannot last long hence not applicable in specific applications. Here, we established a three-dimensional microchamber embedded with two pairs of microelectrodes to investigate the rotational phenomenon of bulk of liquid which we called 'water fan' effect based on the electrohydrodynamics force. When proper voltages were applied on these microelectrodes, the tornado-like rotation would be generated. Both the numerical and experimental results showed that the controllable and continuous rotation could be achieved in the microchamber. In addition, the concentration effect resulting from the rotation flow was also observed. The proposed method offers great promises in providing theoretical and practical guideline in microfluidic devices for mixing, separating, and cooling applications.
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Current food production faces tremendous challenges from growing human population, maintaining clean resources and food qualities, and protecting climate and environment. Food sustainability is mostly a cooperative effort resulting in technology development supported by both governments and enterprises. Multiple attempts have been promoted in tackling challenges and enhancing drivers in food production. Biosensors and biosensing technologies with their applications, are being widely applied to tackling top challenges in food production and its sustainability. Consequently, a growing demand in biosensing technologies exists in food sustainability. Microfluidics represents a technological system integrating multiple technologies. Nanomaterials, with its technology in biosensing, is thought to be the most promising tool in dealing with health, energy, and environmental issues closely related to world populations. The demand of point of care (POC) technologies in this area focus on rapid, simple, accurate, portable, and low-cost analytical instruments. This review provides current viewpoints from the literature on biosensing in food production, food processing, safety and security, food packaging and supply chain, food waste processing, food quality assurance, and food engineering. The current understanding of progress, solution, and future challenges, as well as the commercialization of biosensors are summarized.
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Técnicas Biossensoriais/métodos , Análise de Alimentos/métodos , Animais , Técnicas Biossensoriais/instrumentação , Análise de Alimentos/instrumentação , Embalagem de Alimentos/instrumentação , Embalagem de Alimentos/métodos , Qualidade dos Alimentos , Humanos , Sistemas Automatizados de Assistência Junto ao LeitoRESUMO
Infectious bronchitis virus (IBV), an avian coronavirus, significantly affects the performance of both the egg-laying and meat-type birds causing the foremost of economic loss in poultry industry. This paper aims to develop a rapid, low-cost, and sensitive biosensor for IBV detection by using molybdenum disulfide (MoS2). MoS2 is a 2-D nanosheet which has strong high fluorescence-quenching ability when applied to a dye-labeled antibody (Ab). In this paper, we developed an Ab-functionalized MoS2-based fluorescent immunosensor, which utilized the fluorescence resonance energy transfer (FRET) between the MoS2 and fluorescence dye during the Ab-antigen interaction. The assay was performed on a low-cost cotton thread-based microfluidic platform due to the good wicking property and flexibility. Upon the optimization of assay conditions, the immunosensor demonstrated remarkable sensitivity of [Formula: see text] EID50 per mL and specificity with a dynamic linear response range of 102-106 EID50 per mL for IBV standard solutions. The developed immunoassay successfully detected the IBV spiked chicken serum with satisfactory results. The foregoing presents its potential application for on-farm detection.
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The risks associated with the presence of hidden allergens in food have increased the need for rapid, sensitive, and reliable methods for tracing food allergens in commodities. Conventional enzyme immunosorbent assay (ELISA) has usually been performed in a centralized lab, requiring considerable time and sample/reagent consumption and expensive detection instruments. In this study, a microfluidic ELISA platform combined with a custom-designed optical sensor was developed for the quantitative analysis of the proteins wheat gluten and Ara h 1. The developed microfluidic ELISA biosensor reduced the total assay time from hours (up to 3.5 h) to 15-20 min and decreased sample/reagent consumption to 5-10 µL, compared to a few hundred microliters in commercial ELISA kits, with superior sensitivity. The quantitative capability of the presented biosensor is a distinctive advantage over the commercially available rapid methods such as lateral flow devices (LFD) and dipstick tests. The developed microfluidic biosensor demonstrates the potential for sensitive and less-expensive on-site determination for rapidly detecting food allergens in a complex sample system.
Assuntos
Alérgenos/análise , Técnicas Biossensoriais , Ensaio de Imunoadsorção Enzimática , Alimentos/efeitos adversos , Microfluídica/métodos , Antígenos de Plantas/análise , Glutens/análise , Glicoproteínas/análise , Dispositivos Lab-On-A-Chip , Proteínas de Membrana , Proteínas de Plantas/análiseRESUMO
The increasing prevalence of food allergies and the intake of packing foods in the past two decades urge the need for more rapid, accurate, and sensitive assays to detect potential allergens in food in order to control the allergen content. Most of the commercial analytical tools for allergen detection rely on immunoassays such as ELISA. As far as disadvantages, ELISA can be time-consuming and expensive. Biosensors appear as a suitable alternative for the detection of allergens because they are rapid, highly sensitive, selective, less expensive, environmentally friendly, and easy to handle. In this study, we developed a microfluidic system integrated with a quantum dots (Qdots) aptamer functionalized graphene oxide (GO) nano-biosensor for simple, rapid, and sensitive food allergen detection. The biosensor utilized Qdots-aptamer-GO complexes as probes to undergo conformational change upon interaction with the food allergens, resulting in fluorescence changes due to the fluorescence quenching and recovering properties of GO by adsorption and desorption of aptamer-conjugated Qdots. This one-step 'turn on' homogenous assay in a ready-to-use microfluidic chip took ~10min to achieve a quantitative detection of Ara h 1, one of the major allergens appearing in peanuts. The results suggested this system had remarkable sensitivity and selectivity. The integration of a microfluidics platform in a homemade miniaturized optical analyzer provides a promising way for the rapid, cost-effective, and accurate on-site determination of food allergens. This biosensor can also be extended to the detection of other food allergens with a selection of corresponding aptamers.
Assuntos
Antígenos de Plantas/análise , Aptâmeros de Nucleotídeos/química , Arachis/química , Técnicas Biossensoriais/instrumentação , Glicoproteínas/análise , Grafite/química , Técnicas Analíticas Microfluídicas/instrumentação , Proteínas de Plantas/análise , Pontos Quânticos/química , Desenho de Equipamento , Análise de Alimentos/instrumentação , Limite de Detecção , Proteínas de Membrana , Óxidos/química , Pontos Quânticos/ultraestrutura , Espectrometria de Fluorescência/instrumentaçãoRESUMO
BACKGROUND: Antimicrobial resistance is a great concern in the medical community, as well as food industry. Soy peptides were tested against bacterial biofilms for their antimicrobial activity. A high throughput drug screening assay was developed using microfluidic technology, RAMAN spectroscopy, and optical microscopy for rapid screening of antimicrobials and rapid identification of pathogens. METHODS: Synthesized PGTAVFK and IKAFKEATKVDKVVVLWTA soy peptides were tested against Pseudomonas aeruginosa and Listeria monocytogenes using a microdilution assay. Microfluidic technology in combination with Surface Enhanced RAMAN Spectroscopy (SERS) and optical microscopy was used for rapid screening of soy peptides, pathogen identification, and to visualize the impact of selected peptides. RESULTS: The PGTAVFK peptide did not significantly affect P. aeruginosa, although it had an inhibitory effect on L. monocytogenes above a concentration of 625 µM. IKAFKEATKVDKVVVLWTA was effective against both P. aeruginosa and L. monocytogenes above a concentration of 37.2 µM. High throughput drug screening assays were able to reduce the screening and bacterial detection time to 4 h. SERS spectra was used to distinguish the two bacterial species. CONCLUSIONS: PGTAVFK and IKAFKEATKVDKVVVLWTA soy peptides showed antimicrobial activity against P. aeruginosa and L. monocytogenes. Development of high throughput assays could streamline the drug screening and bacterial detection process. GENERAL SIGNIFICANCE: The results of this study show that the antimicrobial properties, biocompatibility, and biodegradability of soy peptides could possibly make them an alternative to the ineffective antimicrobials and antibiotics currently used in the food and medical fields. High throughput drug screening assays could help hasten pre-clinical trials in the medical field.
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Pseudomonas aeruginosa is a particularly problematic opportunistic pathogen due to its capacity to form recalcitrant biofilm structures, while cohabiting with other harmful/pathogenic species and harboring the capability to release toxins that cause tissue necrosis. Although it is now recognized that the majority of biofilm infections are polymicrobial, little is known about the complex interactions that occur within polymicrobial communities and few tools exist for studying these interactions. In this study, we have designed a microfluidic model that mimics the relevant physiological properties of wound microenvironment, while incorporating materials present in the human extracellular matrix/wound environment. Using microfluidics combined with imaging techniques, we have validated the robustness of our model comparing traditional GFP-tagging to new fluorescent staining techniques to visualize/resolve individual species within a polymicrobial habitat. We have also demonstrated that chemotactic stimuli may be incorporated into our model through specialized ports in our chamber. Our system is specifically designed for use with high resolution imaging techniques, allowing for data collection throughout the life of the biofilm and in real-time. Ultimately, this model can be used to investigate the spatio-temporal mechanobiological structures of the wound environment, and the response of the bacteria to the drug transport which will significantly contribute to our understanding of the development and progression of polymicrobial biofilm infections.
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Biofilmes/crescimento & desenvolvimento , Coinfecção/microbiologia , Dispositivos Lab-On-A-Chip , Modelos Teóricos , Pseudomonas aeruginosa/fisiologia , Infecção dos Ferimentos/microbiologia , Ferimentos e Lesões/microbiologia , Humanos , Microfluídica , Imagem Óptica , Pseudomonas aeruginosa/crescimento & desenvolvimentoRESUMO
Early detection of dairy animal health issues allows the producer or veterinarian to intervene before the animals' production levels, or even survival, is threatened. An increased concentration of ß-hydroxybutyrate (ßHBA) is a key biomarker for diagnosis of subclinical ketosis (SCK), and provides information on the health stress in cows well before any external symptoms are observable. In this study, quantum dots (QDs) modified with cofactor nicotinamide adenine dinucleotide (NAD(+)) were prepared for the sensing event, by which the ßHBA concentration in the cow's blood and milk samples was determined via fluorescence analysis of the functionalized QDs. The detection was performed on a custom designed microfluidic platform combining with a low cost and miniaturized optical sensor. The sensing mechanism was first validated by a microplate reader method and then applied to the microfluidic platform. Standard ßHBA solution, ßHBA in blood and milk samples from cows were successfully measured by this novel technology with a detection limit at a level of 35 µM. Side by side comparison of the developed microfluidic biosensor with a commercial kit presented its good performance.
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Ácido 3-Hidroxibutírico/análise , Ácido 3-Hidroxibutírico/sangue , Técnicas Biossensoriais/instrumentação , Doenças dos Bovinos/diagnóstico , Cetose/veterinária , Leite/química , Animais , Bovinos , Doenças dos Bovinos/sangue , Desenho de Equipamento , Feminino , Cetose/sangue , Cetose/diagnóstico , Limite de Detecção , Técnicas Analíticas Microfluídicas/instrumentação , NAD/química , Pontos Quânticos/químicaRESUMO
BACKGROUND: Determination of ß-hydroxybutyrate (ßHBA) is a gold standard for diagnosis of Subclinical Ketosis (SCK), a common disease in dairy cows that causes significant economic loss. Early detection of SCK can help reduce the risk of the disease progressing into clinical stage, thus minimizing economic losses on dairy cattle. Conventional laboratory methods are time consuming and labor-intensive, requiring expensive and bulky equipment. Development of portable and robust devices for rapid on-site SCK diagnosis is an effective way to prevent and control ketosis and can significantly aid in the management of dairy animal health. Microfluidic technology provides a rapid, cost-effective way to develop handheld devices for on-farm detection of sub-clinical ketosis. In this study, a highly sensitive microfluidics-based biosensor for on-site SCK diagnosis has been developed. RESULTS: A rapid, low-cost microfluidic biosensor with high sensitivity and specificity was developed for SCK diagnosis. Determination of ßHBA was employed as the indicator in the diagnosis of SCK. On-chip detection using miniaturized and cost-effective optical sensor can be finished in 1 minute with a detection limit of 0.05 mM concentration. Developed microfluidic biosensor was successfully tested with the serum samples from dairy cows affected by SCK. The results of the developed biosensor agreed well with two other laboratory methods. The biosensor was characterized by high sensitivity and specificity towards ßHBA with a detection limit of 0.05 mM. CONCLUSIONS: The developed microfluidic biosensor provides a promising prototype for a cost-effective handheld meter for on-site SCK diagnosis. By using microfluidic method, the detection time is significantly decreased compared to other laboratory methods. Here, we demonstrate a field-deployable device to precisely identify and measure subclinical ketosis by specific labeling and quantification of ß-hydroxybutyate in cow blood samples. A real-time on-site detection system will maximize convenience for the farmers.
Assuntos
Ácido 3-Hidroxibutírico/sangue , Técnicas Biossensoriais/métodos , Doenças dos Bovinos/diagnóstico , Cetose/veterinária , Microfluídica/métodos , Ácido 3-Hidroxibutírico/análise , Animais , Bovinos , Desenho de Equipamento , Feminino , Cetose/diagnóstico , Dispositivos Lab-On-A-Chip , Limite de Detecção , Microfluídica/economia , Microfluídica/instrumentação , Sensibilidade e EspecificidadeRESUMO
OBJECTIVE: To assess the correlations between objective measurements of 108 finished orthodontic cases and subjective assessments made by 69 orthodontic specialists, to explore the statistically significant measuring categories of cast and cephalogram and to validate the regression model. METHODS: A stratified random sample of 108 cases was drawn from the large sample of 2 383 patients who finished orthodontic treatment between July 2006 and August 2008 in six orthodontic treatment centers around China. For each patient, the post-treatment information sources evaluated in this study included standardized plaster study casts and a lateral cephalometric X-ray image. These information sources were evaluated both singly and in combination by a panel of 69 orthodontic specialists. The average subjective grading scores of 69 orthodontists were regarded as the gold standard. Six examiners used the peer assessment rating (PAR) index and American board of orthodontics-objective grading system (ABO-OGS) to measure all the study casts respectively and three other examiners measured all the lateral cephalometric X-ray images by using customized software. The objective measuring data were correlated with the gold standard. The correlations between the objective measurement and the subjective evaluation were assessed, the statistically significant measuring categories of cast and cephalogram were explored and the regression model was validated. RESULTS: The ABO-OGS scores of "occlusal relationship" correlated most strongly with the subjective scores of cast (r=0.655, P<0.01), and the secondarily correlated category with those were the PAR scores of "overjet" (r=0.525, P<0.01). The proclination of the lower incisors correlated most strongly with the subjective scores of cephalogram (r=0.446, P<0.01), and the secondarily correlated category with those was the protrusion of the lower lips (r=0.436, P<0.01). Nine components were predictive for the post-treatment model and lateral ephalometric film (Post-M+C) outcome: alignment (ABO-OGS), occlusal relationship (ABO-OGS), interproximal contact(ABO-OGS), L1/NB°, overjet (PAR), SNB°, occlusal contacts (ABO-OGS), U1/SN2° and centerline (PAR). These 9 components accounted for 72% of the variability in the average subjective grading scores. CONCLUSION: The objective regression model could replace the averaged opinion of Chinese orthodontic experts effectively, making objective assessment of orthodontic treatment outcome for Chinese patients.