The Subjective and Objective Improvement of Non-Invasive Treatment of Schumann Resonance in Insomnia-A Randomized and Double-Blinded Study.

Purpose: Accumulated studies revealed that electromagnetic field can affect human brain and sleep, and the extremely low-frequency electromagnetic field, Schumann resonance, may have the potential to reduce insomnia symptoms. The purpose of this study was to investigate the responses of patients with insomnia to a non-invasive treatment, Schumann resonance (SR), and to evaluate its effectiveness by subjective and objective sleep assessments. Patients and Methods: We adopted a double-blinded and randomized design and 40 participants (70% female; 50.00 ± 13.38 year) with insomnia completed the entire study. These participants were divided into the SR-sleep-device group and the placebo-device group and were followed up for four weeks. The study used polysomnography (PSG) to measure objective sleep and used sleep diaries, Pittsburgh Sleep Quality Inventory (PSQI), Epworth Sleepiness Scale (ESS), and visual analogy of sleep satisfaction to measure subjective sleep. The 36-Item Short-Form Health Survey (SF-36) was used to evaluate quality of life. Chi-square test, Mann-Whitney U-test, and Wilcoxon test were used to analyze the data. Results: About 70% of the subjects were women, with an average age of 50±13.38 years and an average history of insomnia of 9.68±8.86 years. We found that in the SR-sleep-device group, objective sleep measurements (sleep-onset-latency, SOL, and total-sleep-time, TST) and subjective sleep questionnaires (SOL, TST, sleep-efficiency, sleep-quality, daytime-sleepiness, and sleep-satisfaction) were significantly improved after using the SR-sleep-device; in the placebo-device group, only such subjective sleep improvements as PSQI and sleep-satisfaction were observed. Conclusion: This study demonstrates that the SR-sleep-device can reduce the insomnia symptoms through both objective and subjective tests, with minimal adverse effects. Future studies can explore the possible mechanism of SR and health effects and, with a longer tracking time, verify the effectiveness and side effects.

Effect of extremely low frequency electromagnetic field parameters on the proliferation of human breast cancer.

Extremely low-frequency electromagnetic field (ELF-EMF) exposures influence many biological systems. These effects are mainly related to the intensity, duration, frequency, and pattern of the ELF-EMF. Our intent was to characterize the effect of specific pulsed electromagnetic fields on the in vitro proliferation of MCF-7 adenocarcinoma and MDA-MB-231 breast cancer cell lines and one non-cancerous M10 breast epithelial cell line. The following four important parameters of ELF-EMF were examined: frequencies (7.83 ± 0.3, 23.49 ± 0.3, and 39.15 ± 0.3 Hz), flux density (0.5 and 1 mT), exposure duration (12, 24, and 48 h), and the exposure methodology (continuous exposure versus switching exposure). The viability of MDA-MB-231 cells exposed to the optimized ELF-EMF pattern (7.83 ± 0.3 Hz, 1 mT, and 6 h switching exposure) was 40.1%. By contrast, the optimized ELF-EMF parameters that were most cytotoxic to breast cancer MDA-MB-231 cells were not damaging to normal M10 cells. In vitro studies also showed that exposure of MDA-MB-231 cells to the optimized ELF-EMF pattern promoted Ca2+ influx and resulted in apoptosis. These data confirm that exposure to this specific ELF-EMF pattern can influence cellular processes and inhibit cancer cell growth. The specific ELF-EMF pattern determined in this study may provide a potential anti-cancer treatment in the future.

Inhibition of B16F10 Cancer Cell Growth by Exposure to the Square Wave with 7.83+/-0.3Hz Involves L- and T-Type Calcium Channels.

Extremely low-frequency electromagnetic field (ELF-EMF) exposure influences many biological systems; these effects are mainly related to the intensity, duration, frequency, and pattern of the ELF-EMF. In this study, exposure to square wave with 7.83±0.3 Hz (sweep step 0.1 Hz) was shown to inhibit the growth of B16F10 melanoma tumor cells. In addition, the distribution of the magnetic field was calculated by Biot-Savart Law and plotted using MATLAB. In vitro studies demonstrated a decrease in B16F10 cell proliferation and an increase of Ca2+ influx after 48 h of exposure to the square wave. Ca2+ influx was also partially blocked by inhibition of voltage-gated L- and T-type Ca2+ channels. The data confirmed that the specific time-varying ELF-EMF had an anti-proliferation effect on B16F10 cells and that the inhibition is related to Ca2+ and voltage-gated L- and T-type Ca2+ channels.

Effects of extremely low-frequency electromagnetic fields on B16F10 cancer cells.

This paper presents a method to inhibit B16F10 cancer cells using extremely low-frequency electromagnetic fields (ELF-EMFs) and to evaluate cell viability using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. The study examined the effect of a natural EMF resonance frequency (7.83 Hz) and a power line frequency (60 Hz) on B16F10 cancer cells for 24 and 48 h. The B16F10 cancer cells were also exposed to sweep frequencies in several sweep intervals to quantitatively analyze the viability of cancer cells. The results yielded a 17% inhibition rate under 7.83 Hz compared with that of the control group. Moreover, sweep frequencies in narrow intervals (7.83 ± 0.1 Hz for the step 0.05 Hz) caused an inhibition rate of 26.4%, and inhibitory effects decreased as frequency sweep intervals increased. These results indicate that a Schumann resonance frequency of 7.83 Hz can inhibit the growth of cancer cells and that using a specific frequency type can lead to more effective growth inhibition.

Impedance measurement system for automatic determination of glycated hemoglobin.

In this study, an automatic glycated hemoglobin (HbA1c) impedance measurement system (AHMS) is developed for the detection of HbA1c. The proposed device removes some of the drawbacks of common instruments for HbA1c detection (i.e., large, expensive, difficult to operate) by detecting the ratio of HbA1c to Hb. The method is label-free and requires only a small sample volume; no additional reagents are required. The manpower consumption and bulk of the instrument are also reduced. The method provides a simple way to analyze impedance deviation and effectively reduces the effort required by the operator. The ratios of HbA1c to Hb (4%-7%) are well distinguished, and the experiment is used to build a database for AHMS. To check the reliability of the proposed system, a sample test using three different ratios of HbA1c is applied in this study. The sample test uses HbA1c to Hb ratios of 4.7%, 5.6%, and 6.8%, and the determined experimental values are 4.93%, 5.8%, and 6.83%, respectively. The sample test has an accuracy of approximately 96.99%. Based on these results, the proposed system for detecting HbA1c through protein coverage is both effective and feasible.

Hemodialysis vascular access stenosis detection using auditory spectro-temporal features of phonoangiography.

For end-stage renal disease patients undergoing hemodialysis, thrombosis caused by stenosis hinders the long-term use of vascular access. However, traditional spectral bruit analysis techniques for detecting the severity of vascular access stenosis are not robust. Accordingly, the present study proposes an automated method for mimicking a trained practitioner in performing the auscultation process. In the proposed approach, the bruit obtained using a standard phonoangiographic method is transformed into the time-frequency domain, and two spectro-temporal features, namely the auditory spectrum flux and the auditory spectral centroid, are then extracted. The distributions of the two features are analyzed using a multivariate Gaussian distribution (MGD) model. Finally, the distribution parameters of the MGD model are used to detect the presence (or otherwise) of vascular access stenosis. The validity of the proposed approach is investigated using the phonoangiography signals obtained from 16 hemodialysis patients with straight arteriovenous grafts over the upper arm region. The results show that the MGD covariance matrix coefficient of the auditory spectral centroid feature yields an accuracy of 83.87 % in detecting significant vascular access stenosis. Thus, the proposed method has significant potential for the applications of vascular access stenosis detection.

Battery-powered portable instrument system for single-cell trapping, impedance measurements, and modeling analyses.

A battery-powered portable instrument system for the single-HeLa-cell trapping and analyses is developed. A method of alternating current electrothermal (ACET) and DEP are employed for the cell trapping and the method of impedance spectroscopy is employed for cell characterizations. The proposed instrument (160 mm × 170 mm × 110 mm, 1269 g) equips with a highly efficient energy-saving design that promises approximately 120 h of use. It includes an impedance analyzer performing an excitation voltage of 0.2-2 Vpp and a frequency sweep of 11-101 kHz, function generator with the sine wave output at an operating voltage of 1-50 Vpp with a frequency of 4-12 MHz, cell-trapping biochip, microscope, and input/output interface. The biochip for the single cell trapping is designed and simulated based on a combination of ACET and DEP forces. In order to improve measurement accuracy, the curve fitting method is adopted to calibrate the proposed impedance spectroscopy. Measurement results from the proposed system are compared with results from a precision impedance analyzer. The trapped cell can be modeled for numerical analyses. Many advantages are offered in the proposed instrument such as the small volume, real-time monitoring, rapid analysis, low cost, low-power consumption, and portable application.

Glycated hemoglobin (HbA1c) affinity biosensors with ring-shaped interdigital electrodes on impedance measurement.

Glycated hemoglobin (HbA1c) is one of the most important diagnostic assays for the long-term mark of glycaemic control in diabetes. This study presents an affinity biosensor for HbA1c detection which is label-free based on the impedance measurement, and it features low cost, low sample volume, and requires no additional reagent in experiments. The ring-shaped interdigital electrodes (RSIDEs) are designed to promote the distribution uniformity and immobilization efficiency of HbA1c, and are further employed to characterize the impedance change and identify various concentrations of HbA1c. The self-assembled monolayer (SAM) of thiophene-3-boronic acid (T3BA) is provided to modify the gold electrode surface. Afterwards, the esterification reaction between HbA1c and T3BA produces a relative change of electrical property on the electrode surface. The RSIDEs with SAM of T3BA exhibit a wide range from 100 to 10 ng/µL producing an approximate logarithmic decrease of impedance, a low detection limit of 1 ng/µL, a good selectivity and short-term stability for HbA1c determination. The remarkable advantages (miniaturization and low-cost) fill the bill of point-care diagnostics for portable sensor development.

Adjustable trapping position for single cells using voltage phase-controlled method.

We present an advanced technique improving upon the micron-sized particle trap integrated in biochip systems using a planar structure to generate an adjustable trapping position by utilizing voltage phase-controlled (VPC) method and negative dielectrophoresis (nDEP) theory in high conductivity physiological media. The designed planar and split structure is composed of independent components of measuring and trapping micro-electrodes. Through different voltage configurations on the device, the trapped position of single particles/cells was selected and adjusted in vertical and horizontal directions. The numerical simulations verify our theoretical predictions of the effects at the various voltages. It shows that the trapped position can be adjusted in the vertical (0 to 26 µm) and horizontal (0 to 74 µm) directions. In experiments, the single particles/cells is captured, measured, and then released, with the same process being repeated twice to demonstrate the precision of the positioning. The measurement results determined that particles at various heights result in different magnitude values, while the impedance error is less than 5% for the proposed electrode layout. Finally, the experiments are performed to verify that a particle/cell can be precisely trapped on the selected site in both the vertical and horizontal directions.

Influence of electromagnetic signal of antibiotics excited by low-frequency pulsed electromagnetic fields on growth of Escherichia coli.

Energy medicine (EM) provides a new medical choice for patients, and its advantages are the noninvasive detection and nondrug treatment. An electromagnetic signal, a kind of EM, induced from antibiotic coupling with weak, extremely low-frequency pulsed electromagnetic fields (PEMFs) is utilized for investigating the growth speed of Escherichia coli (E. coli). PEMFs are produced by solenoidal coils for coupling the electromagnetic signal of antibiotics (penicillin). The growth retardation rate (GRR) of E. coli is used to investigate the efficacy of the electromagnetic signal of antibiotics. The E. coli is cultivated in the exposure of PEMFs coupling with the electromagnetic signal of antibiotics. The maximum GRR of PEMFs with and without the electromagnetic signal of antibiotics on the growth of E. coli cells in the logarithmic is 17.4 and 9.08%, respectively. The electromagnetic signal of antibiotics is successfully coupled by the electromagnetic signal coupling instrument to affect the growth of E. coli. In addition, the retardation effect on E. coli growth can be improved of by changing the carrier frequency of PEMFs coupling with the electromagnetic signal of antibiotics. GRR caused by the electromagnetic signal of antibiotics can be fixed by a different carrier frequency in a different phase of E. coli growth.

Single HeLa and MCF-7 cell measurement using minimized impedance spectroscopy and microfluidic device.

This study presents an impedance measurement system for single-cell capture and measurement. The microwell structure which utilizes nDEP force is used to single-cell capture and a minimized impedance spectroscopy which includes a power supply chip, an impedance measurement chip and a USB microcontroller chip is used to single-cell impedance measurement. To improve the measurement accuracy of the proposed system, Biquadratic fitting is used in this study. The measurement accuracy and reliability of the proposed system are compared to those of a conventional precision impedance analyzer. Moreover, a stable material, latex beads, is used to study the impedance measurement using the minimized impedance spectroscopy with cell-trapping device. Finally, the proposed system is used to measure the impedance of HeLa cells and MCF-7 cells. The impedance of single HeLa cells decreased from 9.55 × 10(3) to 3.36 × 10(3) Ω and the impedance of single MCF-7 cells decreased from 3.48 × 10(3) to 1.45 × 10(3) Ω at an operate voltage of 0.5 V when the excitation frequency was increased from 11 to 101 kHz. The results demonstrate that the proposed impedance measurement system successfully distinguishes HeLa cells and MCF-7 cells.

Experimental study of dielectrophoresis and liquid dielectrophoresis mechanisms for particle capture in a droplet.

This work presents a microfluidic system that can transport, concentrate, and capture particles in a controllable droplet. Dielectrophoresis (DEP), a phenomenon in which a force is exerted on a dielectric particle when it is subjected to a non-uniform electric field, is used to manipulate particles. Liquid dielectrophoresis (LDEP), a phenomenon in which a liquid moves toward regions of high electric field strength under a non-uniform electric field, is used to manipulate the fluid. In this study, a mechanism of droplet creation presented in a previous work that uses DEP and LDEP is improved. A driving electrode with a DEP gap is used to prevent beads from getting stuck at the interface between air and liquid, which is actuated with an AC signal of 200 V(pp) at a frequency of 100 kHz. DEP theory is used to calculate the DEP force in the liquid, and LDEP theory is used to analyze the influence of the DEP gap. The increment of the actuation voltage due to the electrode with a DEP gap is calculated. A set of microwell electrodes is used to capture a bead using DEP force, which is actuated with an AC signal of 20 V(pp) at a frequency of 5 MHz. A simulation is carried out to investigate the dimensions of the DEP gap and microwell electrodes. Experiments are performed to demonstrate the creation of a 100-nL droplet and the capture of individual 10-µm polystyrene latex beads in the droplet.

Integration of single-cell trapping and impedance measurement utilizing microwell electrodes.

The ability to research individual cells has been seen as important in many kinds of biological studies. In the present study, cell impedance analysis is integrated into a single-cell trapping structure. For the purpose of precise positioning, a cell manipulation and measurement microchip, which uses an alternating current electrothermal effect (ACET) and a negative dielectrophoresis (nDEP) force to move a particle and cell on measurement electrodes, is developed. An ACET and an nDEP can be easily combined with subsequent analyses based on electric fields. A microwell presented in a previous study is separated into two parts, which are regarded as the measurement electrodes. The original structure is modified for precise positioning. Numerical simulations and analyses are conducted to compute and analyze the effects of the structural parameters. The results of simulations and analyses are used to obtain the optimum structure for the cell. The capture range of the microwell can be designed for cells of various sizes. In order to demonstrate the precision of the positioning, a particle is captured, measured, and released twice. The results show that the impedance error of the particle is about 3%. Finally, the developed structure is applied to trap and measure the impedance of a HeLa cell.

Effects of electrode geometry and cell location on single-cell impedance measurement.

Measurements on single cells provide more accurate and in-depth information about electrical properties than those on pathological tissues. The relationship between electrode geometry and the location of a cell on microfluidic devices greatly affects the accuracy of single-cell impedance measurement. Accordingly, this study presents numerical solutions from the FEM simulation of the COMSOL multiphysics package and experimental measurements to analyze the effects of electrode geometry and cell location on microfluidic devices. An equivalent electrical circuit model is developed to obtain the impedance and sensitivity of various cell locations on various electrode geometries using FEM simulation. According to the simulation results, the parallel electrodes have the largest sensing area (39 microm(2)) and the highest sensitivity (0.976) at a voltage of 0.1 V and a frequency of 100 kHz. Increasing the width of electrodes provides a large sensing area but reduces sensitivity, whereas decreasing the gap between electrodes increases both sensing area and sensitivity. In experiments, the results demonstrate that the magnitude is inversely proportional to the overlap area of the cell and electrodes. Moreover, the impedance of single HeLa cells measured at various cell locations can be modified using equations determined from the modeling and experimental results.

Single-cell trapping utilizing negative dielectrophoretic quadrupole and microwell electrodes.

The handling of individual cells, which has attracted increasing attention, is a key technique in cell engineering such as gene introduction, drug injection, and cloning technology. Alternating current (AC) electrokinetics has shown great potential for microfluidic functions such as pumping, mixing, and concentrating particles. The non-uniform electric field gives rise to Joule heating and dielectrophoresis (DEP). The motion of particles suspended in the medium can be influenced directly, by means of dielectrophoretic effects, and indirectly, via fluid flow through a viscous drag force that affects the particles. Thus alternating current electrothermal effect (ACET) induced flow and DEP force can be combined to manipulate and trap single particles and cells. This study presents a microfluidic device which is capable of specifically guiding and capturing single particles and cells by ACET fluid flow and the negative dielectrophoretic (nDEP) trap, respectively. The experiment was operated at high frequencies (5-12 MHz) and in a culture medium whose high conductivity (sigma=1.25S/m) is of interest to biochemical analysis and environmental monitoring, which are both prone to producing ACET and nDEP. Manipulation of particle motion using ACET-induced fluid flow to the target trap is modeled numerically and is in good agreement with the experimental results.

Effect of electrode geometry on performance of EWOD device driven by battery-based system.

This study develops a driving system for an electrowetting-on-dielectric (EWOD) device comprising a 9 V battery, an ATmega8535 microprocessor, a DC/DC converter, two regulator ICs and a switch circuit. The driving system greatly improves the portability of the EWOD device and is capable of generating a square wave with voltages ranging from 50~100 V(pp) and frequencies in the range 1~5 kHz. A series of experimental and numerical investigations are performed to investigate the effect of the conducting electrode geometry on the droplet velocity in the EWOD device. Three different electrode configurations are considered, namely a linear array of square electrodes, a series of interdigitated electrodes having either two or three fingers, and a series of interdigitated electrodes having five or six fingers. The experimental results show that the corresponding droplet velocities are 7.25 mm/s, 8.17 mm/s and 7.82 mm/s, respectively. The simulation results indicate that the pressure difference induced within the droplets actuated by the square, interdigitated (2323) and interdigitated (5656) electrodes has a value of 15.5 N/m², 262 N/m² and 141.1 N/m², respectively. The corresponding droplet velocities are 33.8 mm/s, 72.7 mm/s and 64.5 mm/s, respectively. Overall, the experimental and numerical results indicate that the interdigitated (2323) electrode optimizes the transportation of the droplets in the EWOD device. The improved droplet velocity obtained using this particular electrode configuration is attributed to an increased length of the contact line between the droplet and the actuating electrode, which in turn increases the driving force.

A systematic investigation into the electrical properties of single HeLa cells via impedance measurements and COMSOL simulations.

The electrical properties of single cells provide fundamental insights into their pathological condition and are therefore of immense interest to medical practitioners. Accordingly, this study captures single HeLa cells using a microfluidic device and then measures their impedance properties using a commercial impedance spectroscopy system. The experimental system is modeled by an equivalent electrical circuit and COMSOL simulations are then performed to establish the conductivity, permittivity and impedance of single HeLa cells under various operational frequencies and voltages. At an operational voltage of 0.2 V, the maximum deviation between the experimental and simulation results for the magnitude and phase of the HeLa cell impedance is found to be 9.5% and 4.2%, respectively. In general, both sets of results show that the conductivity and permittivity of single HeLa cells increase with an increasing operational voltage. Moreover, an increasing frequency is found to increase the conductivity of HeLa cells at all values of the operational voltage, but to reduce the permittivity for operational voltages in the range 0.6-1.0 V. Based upon the simulation and experimental results, empirical equations are constructed to predict the conductivity and permittivity of single HeLa cells under specified values of the operational voltage and frequency, respectively. The maximum discrepancy between the predicted results and the simulation results for the permittivity and conductivity of the HeLa cells at an operational voltage of 0.2 V is found to be just 0.5% and 4.5%, respectively.

Fabrication of protein chips based on 3-aminopropyltriethoxysilane as a monolayer.

Although 3-aminopropyltriethoxysilane (APTES) is widely adopted as a monolayer in biosensors, experimental silanization takes at least 1 h at high temperature. Therefore, the feasibility of the silanization with APTES in a short reaction time and at room temperature was investigated. The surface modification of glass slides using a self-assembled monolayer of APTES with a concentration of 10% was studied by immobilizing FITC. APTES was successfully immobilized on the glass slide. The effect of reaction temperature and time of silanization were investigated. Various silanization conditions of APTES were examined by contact angle measurement and fluorescence microscopy. The surface of glass patterns with a gold thin film as background was characterized by determining the fluorescent intensities following the immobilization of fluorescein isothiocyanate (FITC), protein A-FITC, antimouse IgG-FITC and sheep anti-bovine albumin-FITC. The normalized fluorescent intensity indicated that a short period (4 min) of silanization at 25 degrees C suffices to form an APTES thin film by the immobilization of protein A on a glass surface. Such a condition does not require microheaters and temperature sensors in a microfluidic system, which will significantly reduce the manufacturing process, cost, and reaction time in the future.

Effect of actuation sequence on flow rates of peristaltic micropumps with PZT actuators.

Many biomedical applications require the administration of drugs at a precise and preferably programmable rate. The flow rate generated by the peristaltic micropumps used in such applications depends on the actuation sequence. Accordingly, the current study performs an analytical and experimental investigation to determine the correlation between the dynamic response of the diaphragms in the micropump and the actuation sequence. A simple analytical model of a peristaltic micropump is established to analyze the shift in the resonant frequency of the diaphragms caused by the viscous damping effect. The analytical results show that this damping effect increases as the oscillation frequency of the diaphragm increases. A peristaltic micropump with three piezoelectric actuators is fabricated on a silicon substrate and is actuated using 2-, 3-, 4- and 6-phase actuation sequences via a driving system comprising a microprocessor and a phase controller. A series of experiments is conducted using de-ionized water as the working fluid to determine the diaphragm displacement and the flow rates induced by each of the different actuation sequences under phase frequencies ranging from 50 Hz to 1 MHz. The results show that the damping effect of actuation sequences influences diaphragm resonant frequency, which in turn affects the profiles of flow rates.

Modified fabrication process of protein chips using a short-chain self-assembled monolayer.

In previous work a short chain SAM, 4,4-Dithiodibutyric Acid (DTBA) was found to be a thin monolayer in protein chips. However, obtaining uniform fluorescent intensity remains difficult because water-soluble carbodiimides (EDC) in an aqueous system cause the hydrolysis of N-hydroxysuccinimide ester (NHS esters). The hydrolysis of NHS esters reduces coupling yields and therefore reduces the fluorescent intensity of protein chips. The NHS can increase the stability of active intermediate resulting from the reaction of EDC and NHS, but the ratio of the concentration of EDC to that of NHS strongly affects this stability. The effects of the solvents used in the washing step are studied to solve this problem. The results reveal that PBST (PBS + 5% Tween20) is more effective in reducing the hydrolysis of NHS esters than deionized water. Additionally, the effects of 3:1 and 5:2 EDC/NHS ratios on the chips are examined. The 3:1 EDC/NHS ratio yields a higher fluorescent intensity than the 5:2 ratio. The effects on the chips of dissolving EDC in DI water, DI water + 0.1 M MES and alcohol are also investigated. The results show that alcohol provides higher fluorescent intensity than other solvents and the reaction time of 4 h yields a high fluorescent intensity with 3:1 EDC/NHS ratio. A modified fabrication process of protein chips using 4,4-DTBA is developed. In this work, 160 mM 4,4-DTBA is used as a self-assembled monolayer in the fabrication of protein chips. Experiments to characterize 4,4-DTBA are performed by contact angle goniometry and Fourier transform infrared spectroscopy (FTIR). Furthermore, the immobilized protein A-FITC (fluorescein isothiocyanate) is adopted in fluorescent assays.