Jida Xing

Enhancing multimodality functional and molecular imaging using glucose-coated gold nanoparticles

AIM To describe how pegylated glucose-coated gold nanoparticles (PEG-Glu-GNPs) can help improve computed tomography (CT) imaging. MATERIALS AND METHODS PEG-Glu-GNPs were designed for use as an imaging nanoprobe to act an effective contrast agent for both CT and PET scans. Twelve BALB/c mice were divided into two groups: mice with injected with PEG-Glu-GNPs and control mice. The mice were examined using high-resolution micro-CT at different time intervals (24 h, 7 days, and 15 days) after the injection of the particles. Greyscale density and CT attenuation values were determined to trace the excretion of the particles over time. RESULTS Tumour contours were easily distinguished from surrounding tissue in mice injected with PEG-Glu-GNPs but not controls. This distinction was still visible at 7 days, but not at 15 days post-injection. CONCLUSION Molecular imaging technology has enabled the development of a new generation of imaging probes. These sophisticated probes can visualize biological processes or enable early diagnosis of diseases in vivo. Compared to conventional CT images and PET scans, PEG-Glu-GNPs significantly improved image quality at the cellular and molecular level, which can significantly aid the early detection of cancer or cancer metastases.

Algal Cell Response to Pulsed Waved Stimulation and Its Application to Increase Algal Lipid Production

Generating renewable energy while sequestering CO2 using algae has recently attracted significant research attention, mostly directing towards biological methods such as systems biology, genetic engineering and bio-refining for optimizing algae strains. Other approaches focus on chemical screening to adjust culture conditions or culture media. We report for the first time the physiological changes of algal cells in response to a novel form of mechanical stimulation, or a pulsed wave at the frequency of 1.5 MHz and the duty cycle of 20%. We studied how the pulsed wave can further increase algal lipid production on top of existing biological and chemical methods. Two commonly used algal strains, fresh-water Chlorella vulgaris and seawater Tetraselmis chuii, were selected. We have performed the tests in shake flasks and 1 L spinner-flask bioreactors. Conventional Gravimetric measurements show that up to 20% increase for algal lipid could be achieved after 8 days of stimulation. The total electricity cost needed for the stimulations in a one-liter bioreactor is only one-tenth of a US penny. Gas liquid chromatography shows that the fatty acid composition remains unchanged after pulsed-wave stimulation. Scanning electron microscope results also suggest that pulsed wave stimulation induces shear stress and thus increases algal lipid production.

Design of a Thermoacoustic Sensor for Low Intensity Ultrasound Measurements Based on an Artificial Neural Network

In therapeutic ultrasound applications, accurate ultrasound output intensities are crucial because the physiological effects of therapeutic ultrasound are very sensitive to the intensity and duration of these applications. Although radiation force balance is a benchmark technique for measuring ultrasound intensity and power, it is costly, difficult to operate, and compromised by noise vibration. To overcome these limitations, the development of a low-cost, easy to operate, and vibration-resistant alternative device is necessary for rapid ultrasound intensity measurement. Therefore, we proposed and validated a novel two-layer thermoacoustic sensor using an artificial neural network technique to accurately measure low ultrasound intensities between 30 and 120 mW/cm2. The first layer of the sensor design is a cylindrical absorber made of plexiglass, followed by a second layer composed of polyurethane rubber with a high attenuation coefficient to absorb extra ultrasound energy. The sensor determined ultrasound intensities according to a temperature elevation induced by heat converted from incident acoustic energy. Compared with our previous one-layer sensor design, the new two-layer sensor enhanced the ultrasound absorption efficiency to provide more rapid and reliable measurements. Using a three-dimensional model in the K-wave toolbox, our simulation of the ultrasound propagation process demonstrated that the two-layer design is more efficient than the single layer design. We also integrated an artificial neural network algorithm to compensate for the large measurement offset. After obtaining multiple parameters of the sensor characteristics through calibration, the artificial neural network is built to correct temperature drifts and increase the reliability of our thermoacoustic measurements through iterative training about ten seconds. The performance of the artificial neural network method was validated through a series of experiments. Compared to our previous design, the new design reduced sensing time from 20 s to 12 s, and the sensor’s average error from 3.97 mW/cm2 to 1.31 mW/cm2 respectively.

Applications of low-intensity pulsed ultrasound to increase monoclonal antibody production in CHO cells using shake flasks or wavebags.

Many technologies, such as cell line screening and host cell engineering, culture media optimization and bioprocess optimization, have been proposed to increase monoclonal antibody (mAb) production in Chinese Hamster Ovary (CHO) cells. Unlike the existing biochemical approaches, we investigated stimulation using low-intensity pulsed ultrasound (LIPUS) as a purely physical approach, offering enhanced scalability, contamination control and cost-efficiency, while demonstrating significantly increased cell growth and antibody production. It was found that daily ultrasound treatments at 40 mW/cm(2) for 5 min during cell culture increased the production of human anti-IL-8 antibody by more than 30% using 10 or 30 mL shake flasks. Further increasing the ultrasound dosage (either intensities or the treatment duration) did not appreciably increase cell growth or antibody production, however feeding the culture with additional highly-concentrated nutrients, glucose and amino acids (glutamine in this case), did further increase cell growth and antibody titer to 35%. Similar ultrasound treatments (40 mW/cm(2), 5 min per day) when scaled up to larger volume wavebags, resulted in a 25% increase in antibody production. Increased antibody production can be attributed to both elevated cell count and the ultrasound stimulation. Theoretical study of underlying mechanisms was performed through the simulations of molecular dynamics using the AMBER software package, with results showing that LIPUS increases cell permeability. The significance of this study is that LIPUS, as a physical-based stimulation approach, can be externally applied to the cell culture without worrying about contamination. By combining with the existing technologies in antibody production, LIPUS can achieve additional mAb yields. Because it can be easily integrated with existing cell culture apparatuses, the technology is expected to be more acceptable by the end users.

Design and Characterization of a Close-Proximity Thermoacoustic Sensor

Although the radiation force balance is the gold standard for measuring ultrasound intensity, it cannot be used for real-time monitoring in certain settings, for example, bioreactors or in the clinic to measure ultrasound intensities during treatment. Foreseeing these needs, we propose a close-proximity thermoacoustic sensor. In this article, we describe the design, characterization, testing and implementation of such a sensor. We designed a 20-mm-diameter plexiglass sensor with a 2-mm-long absorber and tested it against low-intensity pulsed ultrasound generated at a 1.5-MHz frequency, 20% duty cycle, 1-kHz pulse repetition frequency and intensities between 30 and 120 mW/cm(2). The sensor captures the beam, converts the ultrasound power into heat and indirectly measures the spatial-average time-average ultrasound intensity (Isata) by dividing the calculated power by the beam cross section (or the nominal area of the transducers). A thin copper sheet was attached to the back face of the sensor with thermal paste to increase heat diffusivity 1000-fold, resulting in uniform temperature distribution across the back face. An embedded system design was implemented using an Atmel microcontroller programmed with a least-squares algorithm to fit measured temperature-versus-time data to a model describing the temperature rise averaged across the back side of the sensor in relation to the applied ultrasound intensity. After it was calibrated to the transducer being measured, the thermoacoustic sensor was able to measure ultrasound intensity with an average error of 5.46% compared with readings taken using a radiation force balance.

Microelectronic-sensing assay to detect presence of Verotoxins in human faecal samples

Aims:  To develop a novel Vero cell assay that implements a real‐time cell electronic sensing (RT‐CES) system for the determination of the presence of verotoxin‐producing Escherichia coli (VTEC). The assay overcomes the major drawbacks in conventional Vero cell assay, for example, labour‐intensive and time‐consuming.

Increasing vaccine production using pulsed ultrasound waves

Vaccination is a safe and effective approach to prevent deadly diseases. To increase vaccine production, we propose that a mechanical stimulation can enhance protein production. In order to prove this hypothesis, Sf9 insect cells were used to evaluate the increase in the expression of a fusion protein from hepatitis B virus (HBV S1/S2). We discovered that the ultrasound stimulation at a frequency of 1.5 MHz, intensity of 60 mW/cm2, for a duration of 10 minutes per day increased HBV S1/S2 by 27%. We further derived a model for transport through a cell membrane under the effect of ultrasound waves, tested the key assumptions of the model through a molecular dynamics simulation package, NAMD (Nanoscale Molecular Dynamics program) and utilized CHARMM force field in a steered molecular dynamics environment. The results show that ultrasound waves can increase cell permeability, which, in turn, can enhance nutrient / waste exchange thus leading to enhanced vaccine production. This finding is very meaningful in either shortening vaccine production time, or increasing the yield of proteins for use as vaccines.

Pulsed ultrasound for enhancing vaccine production

Hepatitis B is an infectious liver disease and vaccination is an effective way to protect individuals. We have applied mechanical wave stimulation to increase protein production. To validate our design, we used Sf9 insect cells to increase antigen fragment fusion protein expression for hepatitis B virus (HBV S1/S2). We discovered that stimulation at a frequency of 1.5MHz, intensity of 60mW/cm2, for a duration of 10 minutes per day increased HBV S1/S2 production by 15%. This finding is very significant for shortening vaccine production time or increasing the yield of proteins for use as vaccines.

Live demonstration: Mechanical stimulation for increasing algal oil production

Generating renewable energy while sequestering CO2 using algae has recently attracted significant research attention, mostly directed towards biological methods such as systems biology, genetic engineering and bio-refining for optimizing algae strains. Other approaches focus on chemical screening to adjust culture conditions or culture media. We report for the first time the physiological changes of algal cells in response to a novel form of mechanical stimulation, or a pulsed wave at the frequency of 1.5MHz and the duty cycle of 20%. Most importantly, we studied how the pulsed wave can further increase algal oil production on top of existing biological and chemical methods. Two commonly used algal strains, fresh-water Chlorella vulgaris and seawater Tetraselmis chuii, were selected. Conventional Gravimetric measurements show that up to 20% increase for T. chuii at the intensity of 80 mW/cm2 and 10% increase for C. vulgaris in algal oil could be achieved after 8 days of stimulation at the intensity of 100 mW/cm2. Gas chromatography / mass spectrometry (GC/MS) shows that the fatty acid composition remains unchanged after pulsed-wave stimulation. Scanning electron microscope results also suggest that pulsed wave stimulation induces shear stress and thus increase algal oil production.

Developing a thermoacoustic sensor adaptive to ambient temperatures

In this paper, a simple and adaptive thermoacoustic sensor was designed to measure Low Intensity Pulsed Ultrasound (LIPUS). Compared to other thermoacoustic sensor designs, our novelty lies in (i) integrating an ultrasound medium layer during the measurement to simplify the complicated set-up procedures and (ii) taking the effect of ambient temperatures into design consideration. After measuring temperature increases with various ambient temperatures under different ultrasound intensities, a relationship among ultrasound intensities, ambient temperatures and coefficients of temporal temperature changes was calculated. Our improved design has made the sensor easy to operate and its performance more accurate and consistent than the thermoacoustic sensor designs without considering ambient temperatures. In all, our improved design greatly enhances the thermoacoustic sensor in practical ultrasound calibration.