A point-of-care electrochemical biosensor for the rapid and sensitive detection of biomarkers in murine models with LPS-induced sepsis.

Sepsis is a life-threatening condition, which is irreversible if diagnosis and intervention are delayed. The response of the immune cells towards an infection triggers widespread inflammation through the production of cytokines, which may result in multiple organ dysfunction and eventual death. Conventional detection techniques fail to provide a rapid diagnosis because of their limited sensitivity and tedious protocol.

Voltage-modulated surface plasmon resonance biosensors integrated with gold nanohole arrays.

Surface plasmon resonance (SPR) has emerged as one of the most efficient and attractive techniques for optical sensors in biological applications. The traditional approach of an EC (electrochemical)-SPR biosensor to generate SPR is by adopting a prism underneath the sensing substrate, and an angular scan is performed to characterize the reflectivity of target analytes. In this paper, we designed and investigated a novel optical biosensor based on a hybrid plasmonic and electrochemical phenomenon.

An AC electrokinetics-based electrochemical aptasensor for the rapid detection of microRNA-155.

Traditional immunosensors are often limited by low sensitivity and long detection times, for they usually depend on passive diffusion-dominated transport of target analytes for the binding reaction with a bio-recognition element such as enzymes, antibodies, and aptamers. Numerous studies rely on electric field manipulation by using alternating current (AC) electrokinetics to enhance the hybridization rate and reduce the hybridization time for faster and more efficient detection. This study demonstrated a rapid electrochemical aptasensor integrated with an AC electroosmotic (ACEO) flow phenomenon for the enhanced target hybridization of microRNA-155 (miR-155).

Simple In-House Fabrication of Microwells for Generating Uniform Hepatic Multicellular Cancer Aggregates and Discovering Novel Therapeutics.

Three-dimensional (3D) cell culture models have become powerful tools because they better simulate the in vivo pathophysiological microenvironment than traditional two-dimensional (2D) monolayer cultures. Tumor cells cultured in a 3D system as multicellular cancer aggregates (MCAs) recapitulate several critical in vivo characteristics that enable the study of biological functions and drug discovery. The microwell, in particular, has emerged as a revolutionary technology in the generation of MCAs as it provides geometrically defined microstructures for culturing size-controlled MCAs amenable for various downstream functional assays.

In situ study of EDC/NHS immobilization on gold surface based on attenuated total reflection surface-enhanced infrared absorption spectroscopy (ATR-SEIRAS).

In this study, the time-dependent reaction between 11-mercaptoundecanoic acid (11-MUA) and 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide/N-hydroxysuccinimide (EDC/NHS) is precisely characterized using surface enhanced infrared absorption spectroscopy (SEIRAS). According to the high correlation between the spectral results of SEIRAS and the electrochemical behavior, it strongly demonstrates that the EDC/NHS reaction would be obviously interfered by phosphate ions in the neutral pH condition (pH = 7.0).

Chemo-photothermal effects of doxorubicin/silica-carbon hollow spheres on liver cancer.

Chemo-photothermal therapy, which exhibits synergistic effects, is more effective than either of the treatments administered alone because of its superior ability to target and destroy cancer cells. An anti-cancer compound (doxorubicin, DOX) was embedded in silica-carbon hollow spheres (SCHSs) using heat and vacuum to integrate multi-therapeutic effects onto one platform and subsequently improve the anti-cancer efficacy. SCHSs were synthesized a surface activation method and its highly porous surface enhanced the loading content of the desired drug.

Aptamer-based sensor for quantitative detection of mercury (II) ions by attenuated total reflection surface enhanced infrared absorption spectroscopy.

A sensing platform based on the attenuated total reflection surface enhanced infrared absorption spectroscopy (ATR-SEIRAS) technique and immobilized aptamer has been proposed herein for the selective detection of mercury ions (Hg). In the proposed platform, 5' thiolated 32-mer DNA probes with methylene blue at the 3' end were immobilized on a thin gold (Au) surface layer. Following Hg ions interacting with T bases of the aptamer, T-Hg-T bonds are formed; resulting in a hairpin-shaped formation of the DNA and a detectable change in the IR absorbance of the sensing interface.

A bead-based immunofluorescence-assay on a microfluidic dielectrophoresis platform for rapid dengue virus detection.

The proof of concept of utilizing a microfluidic dielectrophoresis (DEP) chip was conducted to rapidly detect a dengue virus (DENV) in vitro based on the fluorescence immunosensing. The mechanism of detection was that the DEP force was employed to capture the modified beads (mouse anti-flavivirus monoclonal antibody-coated beads) in the microfluidic chip and the DENV modified with fluorescence label, as the detection target, can be then captured on the modified beads by immunoreaction. The fluorescent signal was then obtained through fluorescence microscopy, and then quantified by ImageJ freeware.

Dielectrophoresis System for Testing Antimicrobial Susceptibility of Gram-Negative Bacteria to β-Lactam Antibiotics.

Gram-negative bacteria (GNBs) are common pathogens causing severe sepsis. Rapid evaluation of drug susceptibility would guide effective antibiotic treatment and promote life-saving. A total of 78 clinical isolates of 13 Gram-negative species collected between April 2013 and November 2013 from two medical centers in Tainan were tested.

Identification of Staphylococcus spp. and detection of mecA by an oligonucleotide array.

Phenotypic identification of coagulase-negative staphylococci (CoNS) is difficult and many staphylococcal species carry mecA. This study developed an array that was able to detect mecA and identify 30 staphylococcal species by targeting the internal transcribed spacer regions. A total of 129 target reference strains (30 species) and 434 clinical isolates of staphylococci were analyzed.

Optimization of the score cutoff value for routine identification of Staphylococcus species by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry.

Staphylococcus species are important pathogens. We evaluated 2 score cutoffs (2.0 and 1.

Characterization of the elastic and viscoelastic properties of dentin by a nanoindentation creep test.

Dentin is the main supporting structure of teeth, but its mechanical properties may be adversely affected by pathological demineralization. The purposes of this study were to develop a quantitative approach to characterize the viscoelastic properties of dentin after de- and re-mineralization, and to examine the elastic properties using a nanoindentation creep test. Dentin specimens were prepared to receive both micro- and nano-indentation tests at wet and dry states.

The photothermal effect of silica-carbon hollow sphere-concanavalin A on liver cancer cells.

Hepatocellular carcinoma (HCC) is one of the most common cancers and causes of death by cancer. Concanavalin A (ConA) lectin can specifically bind to the glycoprotein receptors of HCC, which are produced by the aberrant overexpression of liver cancer cells. ConA was used in the current study to conjugate on silica-carbon hollow spheres (SCHSs) and applied in the thermal ablation therapy of liver cancer cell lines under near-infrared (NIR) laser irradiation.

Ripple structure-generated hybrid electrokinetics for on-chip mixing and separating of functionalized beads.

We present an electrokinetics-based microfluidic platform that is capable of on-chip manipulating, mixing, and separating microparticles through adjusting the interrelated magnitudes of dielectrophoresis and AC electroosmosis. Hybrid electrokinetic phenomenon is generated from an electric field-induced micro-ripple structure made of ultraviolet-curable glue. Size-dependent particle separation and selective removal over the ripple structure is demonstrated successfully.

Rapid detection of dermatophytes and Candida albicans in onychomycosis specimens by an oligonucleotide array.

Background: Onychomycosis is a fungal infection of nails, leading to the gradual destruction of the nail plate. Treatment of onychomycosis may need long-time oral antifungal therapy that can have potential side effects, thus accurate diagnosis of the disease before treatment is important. Culture for diagnosis of onychomycosis is time-consuming and has high false-negative rates.

Quantitative determination of salvinorin A, a natural hallucinogen with abuse liability, in Internet-available Salvia divinorum and endemic species of Salvia in Taiwan.

In recent years, recreational use of Salvia divinorum (Lamiaceae), a herbal drug that contains a hallucinogenic ingredient, salvinorin A, has become a new phenomenon among young drug users. In Taiwan, as in many other countries, dry leaves of S. divinorum and its related concentrated extract products are available via the Internet.

Physicochemical properties and osteogenic activity of radiopaque calcium silicate-gelatin cements.

The purpose of this study is to evaluate the physicochemical properties and in vitro osteogenic activity of radiopaque calcium silicate-gelatin cements. The radiopacity, setting time, working time, flow, diametral tensile strength, pH value, washout resistance and morphology of the cements with gelatin (0, 5 and 10% by weight) were measured, which compared to a popular endodontic material, ProRoot white-colored mineral trioxide aggregate (WMTA). The cell morphology, cell attachment and proliferation, alkaline phosphatase and osteocalcin levels on the cements were measured by culturing the specimens with dental pulp cells.

A capillary dielectrophoretic chip for real-time blood cell separation from a drop of whole blood.

This study proposes a capillary dielectrophoretic chip to separate blood cells from a drop of whole blood (approximately 1 μl) sample using negative dielectrophoretic force. The separating efficiency was evaluated by analyzing the image before and after dielectrophoretic force manipulation. Blood samples with various hematocrits (10%-60%) were tested with varied separating voltages and chip designs.

A filter-like AuNPs@MS SERS substrate for Staphylococcus aureus detection.

An accurate, highly sensitive and rapid identification assay of cells is extremely important in areas such as medical diagnosis, biological research, and environmental monitoring. Laboratory examinations of clinical isolates require time-consuming and complex processes to identify the colony count, with approximately 10(6)-10(8) cells needed for the characterization of strains. In the present study, a highly sensitive SERS filter-like substrate is prepared with AuNPs embedded in mesoporous silica (denoted as AuNPs@MS) synthesized by a simple one-spot method, and an example of its use for the filtration and concentration of analytes from aqueous samples is reported.

Isoflavonoid-Rich Flemingia macrophylla Extract Attenuates UVB-Induced Skin Damage by Scavenging Reactive Oxygen Species and Inhibiting MAP Kinase and MMP Expression.

In this study, we investigated the antioxidant activity and anti-photoaging properties of an extract of Flemingia macrophylla, a plant rich in isoflavonoid content. Pretreatment of fibroblasts with Flemingia macrophylla extract (FME) inhibited elastase activity, promoted the protein expression of type I procollagen, and attenuated the phosphorylation of mitogen-activated protein (MAP) kinase and the protein expression of matrix-metalloproteinase- (MMP-) 1, 3, and 9. The IC50 values were 2.

Rapid (<5 min) identification of pathogen in human blood by electrokinetic concentration and surface-enhanced Raman spectroscopy.

This study reports a novel microfluidic platform for rapid and long-ranged concentration of rare-pathogen from human blood for subsequent on-chip surface-enhanced Raman spectroscopy (SERS) identification/discrimination of bacteria based on their detected fingerprints. Using a hybrid electrokinetic mechanism, bacteria can be concentrated at the stagnation area on the SERS-active roughened electrode, while blood cells were excluded away from this region at the center of concentric circular electrodes. This electrokinetic approach performs isolation and concentration of bacteria in about three minutes; the density factor is increased approximately a thousand fold in a local area of ~5000 μm(2) from a low bacteria concentration of 5 × 10(3) CFU/ml.

A rapid electrochemical biosensor based on an AC electrokinetics enhanced immuno-reaction.

Fluorescent labelling and chromogenic reactions that are commonly used in conventional immunoassays typically utilize diffusion dominated transport of analytes, which is limited by slow reaction rates and long detection times. By integrating alternating current (AC) electrokinetics and electrochemical impedance spectroscopy (EIS), we construct an immunochip for rapid, sensitive, and label-free detection. AC electroosmosis (ACEO) and positive dielectrophoresis (DEP), induced by a biased AC electric field, can rapidly convect and trap the analyte onto an EIS working electrode within a few minutes.

Biopolymer coated gold nanocrystals prepared using the green chemistry approach and their shape-dependent catalytic and surface-enhanced Raman scattering properties.

This study deals with the preparation of multi-shaped nanoscale gold crystals under synthetically simple, green, and efficient conditions using a seed-mediated growth approach in the presence of hyaluronic acid (HA). These highly biocompatible multi-shaped gold nanocrystals were examined to evaluate their catalytic and surface enhanced Raman scattering (SERS) properties. The results show that the size and shape of the nanocrystals are mainly correlated to the amount of seed, seed size, HA concentration, and reaction temperature.