Paper-based mediatorless enzymatic microfluidic biofuel cells.

In this study, eco-friendly and disposable paper-based membraneless microfluidic enzymatic fuel cells (EFCs) were developed without any mediators to reduce the toxicity and cost of EFCs. Glucose oxidase and laccase were immobilized on multi-walled carbon nanotube electrodes to catalyze the redox reaction of glucose and oxygen. Micromachining techniques well-suited for mass production were used to precisely fabricate micro-scale Y-shaped and cross-shaped EFCs. Experimental measurements showed that the concentration of glucose in the fuel solution affects the cell performance, which occurs because the flow speed of the fuel stream decreases as the concentration of glucose increases. The highest performance of power density (104.2 ± 3.35 µW cm-2) and current density (615.6 ± 3.14 µA cm-2) were obtained with the Y-shaped channel configuration at a glucose concentration of 100 mM. This performance is the best of all paper-based single EFCs reported to date. The new paper-based co-laminar flow mediatorless EFC exhibits strong potential to power miniaturized and portable on-site diagnostic devices.

Relationship Between the Gastrointestinal Side Effects of an Anti-Hypertensive Medication and Changes in the Serum Lipid Metabolome.

An earlier study using a rat model system indicated that the active ingredients contained in the anti-hypertensive medication amlodipine (AMD) appeared to induce various bowel problems, including constipation and inflammation. A probiotic blend was found to alleviate intestinal complications caused by the medicine. To gain more extensive insight into the beneficial effects of the probiotic blend, we investigated the changes in metabolite levels using a non-targeted metabolic approach with ultra-performance liquid chromatography-quadrupole/time-of-fligh (UPLC-q/TOF) mass spectrometry. Analysis of lipid metabolites revealed that rats that received AMD had a different metabolome profile compared with control rats and rats that received AMD plus the probiotic blend. In the AMD-administered group, serum levels of phosphatidylcholines, lysophosphatidylcholines, sphingomyelins, triglycerides with large numbers of double bonds, cholesterols, sterol derivatives, and cholesterol esters (all p < 0.05) were increased compared with those of the control group and the group that received AMD plus the probiotic blend. The AMD-administered group also exhibited significantly decreased levels of triglycerides with small numbers of double bonds (all p < 0.05). These results support our hypothesis that AMD-induced compositional changes in the gut microbiota are a causal factor in inflammation.

Age-dependent pulmonary reactivity to house dust mite allergen: a model of adult-onset asthma?

Asthma is a heterogeneous disease differentiated by factors like allergen sensitivity, inflammation, sex, and age at onset. The mouse model is widely used to study the early-life development of allergic asthma. However, age-dependent allergen responses later in life remain relatively understudied and lack a widely accepted model. To differentiate age-dependent responses to the ubiquitous house dust mite (HDM), 3- and 9-mo-old female C57BL/6 mice were randomized into two groups each and exposed to HDM or phosphate-buffered saline (control) via intranasal instillation for sensitization and challenge phases. At 24 h after challenge, all mice underwent pulmonary function testing and methacholine challenge. Bronchoalveolar lavage fluid (BALF) was collected for assessment of cell differentials, and right lung lobes were fixed, sectioned, and stained for histopathology and immunohistochemistry. Both age groups demonstrated strong inflammatory/allergic responses to HDM exposure. However, only 9-mo-old HDM-exposed mice demonstrated significant airway hyperresponsiveness compared with age-matched controls. These HDM-exposed mice also had 1) statistically significant increases in tissue bronchiolitis, perivasculitis, and BALF neutrophilia relative to their younger counterparts and 2) significantly increased extent of immunostaining compared with all other groups. This study presents a potential model for adult-onset asthma, focusing specifically on the atopic, perimenopausal female phenotype. Our findings suggest that lung function declines with age and that the inflammatory profile of this adult subgroup is a mixed, rather than a simple, atopic, Th2 response. This model may enhance our understanding of how age influences the development of asthmic-like symptoms in older subgroups.

Photochemical crosslinking of caries-affected dentin combined with total- or self-etch systems.

PURPOSE: To evaluate the effects of collagen crosslinking with riboflavin 0.1% and ultraviolet-A (UVA) 5.4 J on bond strength of total-etch or self-etch adhesives on caries-affected dentin. METHODS: Sixty human caries-affected molars were randomly divided into three groups: control (C), riboflavin (R), and riboflavin + 3 minutes of UVA (R+UVA). After each treatment, either total-etch or self-etch adhesives were applied following the manufacturer's instructions, and composite stubs were built up on the treated surfaces. They were de-bonded in tension to measure bond strength. Twelve extra molars were used for scanning electron microscope (SEM) analysis. RESULTS: We observed that R+UVA-treated group yielded significantly higher bond strengths for carious dentin when the total-etch adhesive was applied. For the self-etch adhesive, no statistical differences were observed between the three pretreated-groups. CONCLUSION: Our results, for the first time, are suggesting that etching with phosphoric acid potentialized the benefits of R+UVA crosslinking on carious dentin. R+UVA dentinal collagen crosslinking improves bond strength for caries-affected dentin when using a total-etch adhesive, but did not affect it when using a self-etch adhesive.

Design, Fabrication, and Testing of a Microfluidic Device for Thermotaxis and Chemotaxis Assays of Sperm.

Infertile couples needing assisted reproduction are increasing, so a fundamental understanding of motile sperm migration is required. This paper presents an advanced microfluidic device for sperm motion analysis utilizing chemotaxis and thermotaxis simultaneously for the first time. The proposed device is a transparent polydimethylsiloxane- and glass-based microfluidic chip system providing a low-cost, useful, and disposable platform for sperm analysis. The concentration gradient of the chemoattractant (acetylcholine) and the temperature difference are formed along the microchannel. The temperature gradient is generated and controlled by a microheater and microsensor. Thermotactic and chemotactic responses of mouse sperm were examined using the proposed device. Experimental results show that motile mouse sperm are attracted more sensitively under integrated conditions of chemotaxis and thermotaxis rather than individual conditions of chemotaxis and thermotaxis. This sperm analysis device is expected to be a useful tool for the study of mammalian sperm migration and the improvement of artificial insemination techniques.

Development of electrochemical microbiochip for the biological diagnosis of Neisseria gonorrhoeae.

A sexually transmitted disease is an illness that has a high probability of transmission between humans or animals who have sexual contact. Our research is based on the development of a microbiochip for Neisseria gonorrhoeae (N.G.). In our study, we have employed fusion technology between microarray technology and a microfluidic system for quantitative analysis of N.G. A great deal of attention has been focused on electrochemical detection by using a DNA probe, which is a specific DNA sequence and binds to a target biomolecule, because of high affinity, ease of usage, and fast measurement. The microbiochip consisted of two electrode systems and microchannel based PDMS. Our detection principles use electrochemical detection. Consequently, our microbiochip detected 5 ng/mL of N.G. and the correlation rate was over 0.95. We can produce a microbiochip, which could bind to a DNA probe and detect sample of interest. We expect that our electrobiochemical chip will be used for the development of a portable device.

Disposable on-chip microfluidic system for buccal cell lysis, DNA purification, and polymerase chain reaction.

This paper reports the development of a disposable, integrated biochip for DNA sample preparation and PCR. The hybrid biochip (25 × 45 mm) is composed of a disposable PDMS layer with a microchannel chamber and reusable glass substrate integrated with a microheater and thermal microsensor. Lysis, purification, and PCR can be performed sequentially on this microfluidic device. Cell lysis is achieved by heat and purification is performed by mechanical filtration. Passive check valves are integrated to enable sample preparation and PCR in a fixed sequence. Reactor temperature is needed to lysis and PCR reaction is controlled within ±1°C by PID controller of LabVIEW software. Buccal epithelial cell lysis, DNA purification, and SY158 gene PCR amplification were successfully performed on this novel chip. Our experiments confirm that the entire process, except the off-chip gel electrophoresis, requires only approximately 1 h for completion. This disposable microfluidic chip for sample preparation and PCR can be easily united with other technologies to realize a fully integrated DNA chip.

Separation of progressive motile sperm from mouse semen using on-chip chemotaxis.

We present a novel method for the separation of progressive motile sperm from non-progressive motile and immotile sperm. This separation was accomplished by inducing chemotaxis along a longitudinal chemical gradient in a microchip composed of a biocompatible polydimethysiloxane layer and a glass substrate. In a preliminary experiment using fluorescent rhodamine B as a marker, we verified that a chemical gradient was generated by diffusion within the microchannel. We used acetylcholine as a chemoattractant to evaluate the chemotactic response of sperm. We tested the response to a 1/2 to 1/64 dilution series of acetylcholine. The results of a mouse sperm chemotaxis assay showed that progressive motile sperm swam predominantly toward the outlet at an optimal chemical gradient of 0.625 (mg/ml)/mm of acetylcholine. This device provides a convenient, disposable, and high-throughput platform that could function as a progressive motile sperm sorter for potential use in intracytoplasmic sperm injection.

An integrated microfluidic device for rapid cell lysis and DNA purification of epithelial cell samples.

In this paper, we describe the design and fabrication of a microfluidic device for cell lysis and DNA purification, and the results of device tests using a real sample of buccal cells. Cell lysis was thermally executed for two minutes at 80 degrees C in a serpentine type microreactor (20 microL) using an Au microheater with a microsensor. The DNA was then mixed with other residual products and purified by a new filtration process involving micropillars and 50-80 microm microbeads. The entire process of sample loading, cell lysis, DNA purification, and sample extraction was successfully completed in the microchip within five minutes. Sample preparation within the microchip was verified by performing a SY158 gene PCR analysis and gel electrophoresis on the products obtained from the chip. The new purification method enhanced DNA purity from 0.93 to 1.62 after purification.

On-chip immunoassay using surface-enhanced Raman scattering of hollow gold nanospheres.

A surface-enhanced Raman scattering (SERS)-based gradient optofluidic sensor has been developed for a fast and sensitive immunoassay. In this work, a novel microfluidic sensor with functional internal structures has been designed and fabricated. This sensor is composed of three compartments consisting of the gradient channel that serially dilutes the target marker, the injection and mixing area of antibody-conjugated hollow gold nanospheres and magnetic beads, and the trapping area of sandwich immunocomplexes using multiple solenoids. Quantitative analysis of a specific target marker is performed by analyzing its characteristic SERS signals. This SERS-based gradient optofluidic sensor can replace the set of microwells or microtubes used in manual serial dilutions that have been traditionally used in enzyme-linked immunosorbent assay (ELISA)-type assays. The limit of detection for rabbit immunoglobin (IgG) is estimated to be 1-10 ng/mL. This novel SERS-based optofluidic immunoassay system is expected to be a powerful clinical tool for the fast and sensitive medical diagnosis of a disease.

Microchip-based multiplex electro-immunosensing system for the detection of cancer biomarkers.

Microfluidic-based microchips have become the focus of research interest for immunoassays and biomarker diagnostics. This is due to their aptitude for high-throughput processing, small sample volume, and short analysis times. In this paper, we describe the development of a microchip-based multiplex electro-immunosensing system for simultaneous detection of cancer biomarkers using gold nanoparticles and silver enhancer. Our microchip is composed of biocompatible poly(PDMS) and glass substrates. To fix the antibody-immobilized microbeads, we used pillar-type microfilters within a reaction chamber. An immunogold silver staining (IGSS) method was used to amplify the electrical signal that corresponded to the immune complex. To demonstrate this approach, we simultaneously assayed three cancer biomarkers, alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), and prostate-specific antigen (PSA) on the microchip. The electrical signal generated from the result of the immunoreaction was measured and monitored by a PC-based system. The overall assay time was reduced from 3-8 h to about 55 min when compared to conventional immunoassays. The working range of the proposed microchip was from 10(-3) to 10(-1) microg/mL of the target antigen.

A novel microfluidic biosensor based on an electrical detection system for alpha-fetoprotein.

Conventional immunoassays are labor intensive, expensive and time consuming and require large pieces of equipment for detection. Therefore, we have developed and characterized a novel immunoassay methodology comprised of microbeads and microbiochips. In this method, microbeads are used to filter and immobilize antibodies and an immuno-gold silver staining (IGSS) method is then used to amplify electrical signals that correspond to the bound antibodies. The chip used for this system is composed of an inexpensive and biocompatible polydimethylsiloxane (PDMS) layer over a Pyrex glass substrate that contains a platinum (Pt) microelectrode, which is used to detect the electrical signal in this system, the microelectrode is fabricated on the substrate and a microchannel and pillar-type microfilter is formed in the PDMS layer. A sandwich immunoassay approach was applied to detect alpha-fetoprotein (AFP), a cancer biomarker, using this system. The results of this study showed that the time required for a complete assay was reduced by 1h and a detection limit as low as 1 ng/mL was attained when this system used, which indicates that similar bead-based electrical detection systems could be used for the diagnosis of many forms of cancer.