Releasing polysaccharide and protein from yeast cells by ultrasound: Selectivity and effects of processing parameters

A 20 kHz high-intensity ultrasound was employed for the selective release of polysaccharide and protein from yeast cells. While the release of polysaccharide and protein was affected by most of the processing parameters, the release selectivity, which is the ratio of the amount of polysaccharide released to that of protein, designated as T/P value, was only influenced by sonication time, temperature and ionic strength, among which temperature had the greatest influence. The T/P value at 85 °C was a factor of 9.3 of the one at 25 °C. The underlying mechanism of this selectivity is speculated to be thermal denaturation and aggregation of protein within yeast cells at elevated temperatures leading to the decrease of protein release by ultrasound. This finding may be useful in exploring a novel selective process for producing polysaccharide and protein fractions from yeast biomass.     

Sonodynamic inactivation of methicillin-resistant Staphylococcus aureus in planktonic condition by curcumin under ultrasound sonication

Methicillin-Resistant Staphylococcus aureus (MRSA) is an important cause of difficult-to-treat infections. The present study aims to investigate sonodynamic inactivation of MRSA in planktonic condition using curcumin under ultrasound sonication. Dark toxicity of curcumin to MRSA was investigated to choose the concentration range of curcumin used in the study. The uptake of curcumin in MRSA was observed before ultrasound sonication. After sonication colony forming units (CFUs) and bacterial viability were investigated using fluorescence assay. Additionally, chromosomal DNA fragmentation was also analyzed. Curcumin showed no dark toxicity to MRSA in the concentration range of ?500 μM. The maximum uptake of curcumin in MRSA occurred in 50 min after curcumin incubation. Counting of CFUs showed that curcumin had significantly sonodynamic killing effect on MRSA in a curcumin dose-dependent manner, and 5-log reduction in CFU was observed after curcumin treatment (40 μM) at room temperature in the dark for 50 min followed by exposure to ultrasound with intensity of 1.56 W/cm² for 5 min. The ratio of green-fluorescent intensity to red-fluorescent intensity was obviously decreased after curcumin treatment under ultrasound sonication. No significant change in chromosomal DNA was found in the cultured MRSA after the combined treatment of curcumin and ultrasound. These results demonstrated that sonodynamic action of curcumin had significant inactivation of MRSA in planktonic condition.     

Ultrasound treatment enhances cholesterol removal ability of lactobacilli

This study aimed to evaluate the effect of ultrasound treatment on the cholesterol removing ability of lactobacilli. Viability of lactobacilli cells was significantly increased (P < 0.05) immediately after treatment, but higher intensity of 100 W and longer duration of 3 min was detrimental on cellular viability (P < 0.05). This was attributed to the disruption of membrane lipid bilayer, cell lysis and membrane lipid peroxidation upon ultrasound treatment at higher intensity and duration. Nevertheless, the effect of ultrasound on membrane properties was reversible, as the viability of ultrasound-treated lactobacilli was increased (P < 0.05) after fermentation at 37 °C for 20 h. The removal of cholesterol by ultrasound-treated lactobacilli via assimilation and incorporation of cholesterol into the cellular membrane also increased significantly (P < 0.05) upon treatment, as observed from the increased ratio of membrane C:P. Results from fluorescence anisotropies showed that most of the incorporated cholesterol was saturated in the regions of phospholipids tails, upper phospholipids, and polar heads of the membrane bilayer.     

Application of ultrasound as pretreatment for extraction of podophyllotoxin from rhizomes of Podophyllum peltatum

The effect of high-power ultrasound pretreatment on the extraction of podophyllotoxin from Podophyllum peltatum was investigated. Direct sonication by an ultrasound probe horn was applied at 24 kHz and a number of factors were investigated: particle size (0.18-0.6 mm), type of solvent (0-100% aqueous ethanol), ultrasonic treatment time (2-40 min), and power of ultrasound (0-100% power intensity, maximum power: 78 W). The optimal condition of ultrasound was achieved with 0.425-0.6 mm particle size, 10 min sonication time, 35 W ultrasound power, and water as the medium. There was no obvious degradation of podophyllotoxin with ultrasound under the applied conditions, and an improvement in extractability was observed. The SEM microscopic structure change of treated samples disclosed the effect of ultrasound on the tissue cells. The increased pore volume and surface area after ultrasonic treatment also confirmed the positive effect of ultrasound pretreatment on the extraction yield of podophyllotoxin from the plant cells.     

Enhanced growth of lactobacilli and bioconversion of isoflavones in biotin-supplemented soymilk upon ultrasound-treatment

This study aimed at utilizing ultrasound treatment to further enhance the growth of lactobacilli and their isoflavone bioconversion activities in biotin-supplemented soymilk. Strains of lactobacilli (Lactobacillus acidophilus BT 1088, L. fermentum BT 8219, L. acidophilus FTDC 8633, L. gasseri FTDC 8131) were treated with ultrasound (30 kHz, 100 W) at different amplitudes (20%, 60% and 100%) for 60, 120 and 180 s prior to inoculation and fermentation in biotin-soymilk. The treatment affected the fatty acids chain of the cellular membrane lipid bilayer, as shown by an increased lipid peroxidation (P < 0.05). This led to increased membrane fluidity and subsequently, membrane permeability (P < 0.05). The permeabilized cellular membranes had facilitated nutrient internalization and subsequent growth enhancement (P < 0.05). Higher amplitudes and longer durations of the treatment promoted growth of lactobacilli in soymilk, with viable counts exceeding 9 log CFU/mL. The intracellular and extracellular β-glucosidase specific activities of lactobacilli were also enhanced (P < 0.05) upon ultrasound treatment, leading to increased bioconversion of isoflavones in soymilk, particularly genistin and malonyl genistin to genistein. Results from this study show that ultrasound treatment on lactobacilli cells promotes (P < 0.05) the β-glucosidase activity of cells for the benefit of enhanced (P < 0.05) isoflavone glucosides bioconversion to bioactive aglycones in soymilk.     

Potential of Nd:YAG pulsed laser deposition method for coated conductor production

A Nd:YAG laser is environmentally safe and economical with no poisonous or hazardous gases and no expensive gases. We prepared Y123 films by using the fourth harmonic Nd:YAG pulsed laser deposition (PLD) method and optimized the deposition conditions on MgO single crystalline substrates and IBAD-MgO substrates for Y123 coated conductor. We found that the optimal deposition conditions acquired bi-axially aligned Y123 films on both substrates with T_c ∼ 90 K and J_c > 1 MA/cm² at 77 K in self-field. For obtaining high I_c, we fabricated thick Y123 films on both substrates and the maximum I_c per 1 cm in width reached 186 A/cm-width on the IBAD-MgO substrate. Interestingly, there were no a-axis oriented grains within the films up to 1.8 μm thick. This might be an especial feature of the Nd:YAG-PLD method. We believe that the Nd:YAG-PLD method is promising method for RE123 coated conductor production.     

Optimization of ultrasound assisted extraction of antioxidant compounds from marjoram (Origanum majorana L.) using response surface methodology

The present study optimized the ultrasound assisted extraction (UAE) conditions to maximize the antioxidant activity [Ferric ion Reducing Antioxidant Power (FRAP)], total phenol content (TP) and content of individual polyphenols of extracts from marjoram. Optimal conditions with regard to amplitude of sonication (24.4-61.0 μm) and extraction temperature (15-35 °C) and extraction time (5-15 min) were identified using response surface methodology (RSM). The results showed that the combined treatment conditions of 61 μm, 35 °C and 15 min were optimal for maximizing TP, FRAP, rosmarinic acid, luteolin-7-O-glucoside, apigenin-7-O-glucoside, caffeic acid, carnosic acid and carnosol values of the extracts. The predicted values from the developed quadratic polynomial equation were in close agreement with the actual experimental values with low average mean deviations (E%) ranging from 0.45% to 1.55%. The extraction yields of the optimal UAE were significantly (p < 0.05) higher than solid/liquid extracts. Predicted models were highly significant (p < 0.05) for all the parameters studied with high regression coefficients (R²) ranging from 0.58 to 0.989.     

Effects of ultrasound on growth, bioconversion of isoflavones and probiotic properties of parent and subsequent passages of Lactobacillus fermentum BT 8633 in biotin-supplemented soymilk

This study aimed to evaluate the effects of ultrasound on Lactobacillus fermentum BT 8633 in parent and subsequent passages based on their growth and isoflavone bioconversion activities in biotin-supplemented soymilk. The treated cells were also assessed for impact of ultrasound on probiotic properties. The growth of ultrasonicated parent cells increased (P < 0.05) by 3.23-9.14% compared to that of the control during fermentation in biotin-soymilk. This was also associated with enhanced intracellular and extracellular (8.4-17.0% and 16.7-49.2%, respectively; P < 0.05) β-glucosidase specific activity, leading to increased bioconversion of isoflavones glucosides to aglycones during fermentation in biotin-soymilk compared to that of the control (P < 0.05). Such traits may be credited to the reversible permeabilized membrane of ultrasonicated parent cells that have facilitated the transport of molecules across the membrane. The growing characteristics of first, second and third passage of treated cells in biotin-soymilk were similar (P > 0.05) to that of the control, where their growth, enzyme and isoflavone bioconversion activities (P > 0.05) were comparable. This may be attributed to the temporary permeabilization in the membrane of treated cells. Ultrasound affected probiotic properties of parent L. fermentum, by reducing tolerance ability towards acid (pH 2) and bile; lowering inhibitory activities against selected pathogens and reducing adhesion ability compared to that of the control (P < 0.05). The first, second and third passage of treated cells did not exhibit such traits, with the exception of their bile tolerance ability which was inherited to the first passage (P < 0.05). Our results suggested that ultrasound could be used to increase bioactivity of biotin-soymilk via fermentation by probiotic L. fermentum FTDC 8633 for the development of functional food.     

Thin films of polyaniline deposited by MAPLE technique

Polyaniline (PAni) has important electro-conductive properties, high absorbance in microwave range and it is also frequently used in gas sensors because of its capability to convert chemical interactions into electrical signals. The methods of obtaining polyaniline in the form of thin films and/or nanostructures are complicated and request special physical and chemical treatments, both on the substrate surface and for the polymer itself.In this paper we applied matrix assisted pulsed lased evaporation (MAPLE) for obtaining thin films and nanostructures of polyaniline. In MAPLE, the target consisting of the material (usually 0.2-3 wt%) dissolved in a solvent is frozen and it is evaporated using a laser. In our case polyaniline-emeraldine salt (PAni-ES) was dissolved in xylene or toluene, frozen in liquid nitrogen and was used as target. The third and the fourth harmonics of a Nd-YAG laser (λ = 355 nm and 266 nm) were used as laser sources. The obtained films have been characterized by atomic force microscopy, dielectric spectroscopy and Fourier transform infrared spectroscopy. The influence of the solvent type and of the laser parameters (wavelengths and fluence) on the polyaniline structures composition and properties has been investigated.     

Optical properties of microphytobenthic biofilms (MPBOM): Biomass retrieval implication

Microphytobenthos Optical Model (MPBOM) provides the optical properties, absorption coefficient and refractive index, of a laboratory simulated microphytobenthic biofilm using the reflectance measurements derived from HySpex laboratory images, with the final aim of estimating photosynthetically active biomass. The high correlation between this biomass, expressed in chlorophyll a (mg Chl a m⁻²) and the absorption coefficient at the corresponding absorption wavelength of Chl a (673 nm) made possible the estimation of biomass for any absorption coefficient calculated from reflectance measurements of any other data set. The latter was validated for an independent data set which performed an acceptable estimation of biomass in comparison with the biomass measured by HPLC (R²=0.93). Finally, this model is designed to be applied to hyperspectral images, like airborne or satellite, in order to map biomass in the field.     

Preparation and in vitro evaluation of poly(D,L-lactide-co-glycolide) air-filled nanocapsules as a contrast agent for ultrasound imaging

The aim of this study was to prepare air-filled nanocapsules intended ultrasound contrast agents (UCAs) with a biodegradable polymeric shell composed of poly(d,l-lactide-co-glycolide) (PLGA). Because of their size, current commercial UCAs are not capable of penetrating the irregular vasculature that feeds growing tumors. The new generation of UCAs should be designed on the nanoscale to enhance tumor detection, in addition, the polymeric shell in contrast with monomolecular stabilized UCAs improves the mechanical properties against ultrasound pressure and lack of stability. The preparation method of air-filled nanocapsules was based on a modification of the double-emulsion solvent evaporation technique. Air-filled nanocapsules with a mean diameter of 370 ± 96 nm were obtained. Electronic microscopies revealed spherical-shaped particles with smooth surfaces and a capsular morphology, with a shell thickness of ∼50 nm. Air-filled nanocapsules showed echogenic power in vitro, providing an enhancement of up to 15 dB at a concentration of 0.045 mg/mL at a frequency of 10 MHz. Loss of signal for air-filled nanocapsules was 2 dB after 30 min, suggesting high stability. The prepared contrast agent in this work has the potential to be used in ultrasound imaging.     

Effect of sonication on eliminating of phorate in apple juice

The degradation of phorate in apple juice by sonication was investigated in the present study. Results showed that sonication was effective in eliminating phorate in apple juice, and the ultrasonic power and sonication time significantly influenced the degradation of phorate (p < 0.05). The degradation of phorate followed the first-order kinetics model well. Phorate-oxon and phorate sulfoxide were identified as the degradation products of phorate by gas chromatography-mass spectrometry (GC-MS). Moreover, the toxicity of apple juice samples spiked with phorate was significantly reduced by sonication (p < 0.05). The quality indexes of apple juice including pH, titratable acidity (TA), electrical conductivity (EC), total soluble solids (TSS), and the contents of sucrose, glucose and fructose were not affected by sonication, and no visible difference in color was observed between the sonicated samples and the control.     

Investigation of diode parameters using I-V and C-V characteristics of In/SiO2/p-Si (MIS) Schottky diodes

A study on interface states density distribution and characteristic parameters of the In/SiO₂/p-Si (MIS) capacitor has been made. The thickness of the SiO₂ film obtained from the measurement of the corrected capacitance in the strong accumulation region for MIS Schottky diodes was 220 Å. The diode parameters from the forward bias I-V characteristics such as ideality factor, series resistance and barrier heights were found to be 1.75, 106-112 Ω and 0.592 eV, respectively. The energy distribution of the interface state density D_it was determined from the forward bias I-V characteristics by taking into account the bias dependence of the effective barrier height. The interface state density obtained using the I-V characteristics had an exponential growth, with bias towards the top of the valance band, from 9.44×10¹³ eV⁻¹ cm⁻² in 0.329-E_v eV to 1.11×10¹³ eV⁻¹ cm⁻² in 0.527-E_v eV at room temperature. Furthermore, the values of interface state density D_it obtained by the Hill-Coleman method from the C-V characteristics range from 52.9×10¹³ to 1.11×10¹³ eV⁻¹ cm⁻² at a frequency range of 30kHz-1 MHz. These values of D_it and R_s were responsible for the non-ideal behaviour of I-V and C-V characteristics.     

Inactivation of microorganisms by low-frequency high-power ultrasound: 2. A simple model for the inactivation mechanism

A simple theoretical model based on shear forces generated by the collapse of the ultrasound cavities near the surface of a microorganism is proposed. This model requires two parameters which take into account the number of acoustic cavitation bubbles, and the resistance of the cell wall of the microorganism to the shear forces generated by bubble collapse. To validate the model, high-power low frequency (20 kHz) ultrasound was used to inactivate two microorganisms with very different sizes, viz., a bacterium, Enterobacter aerogenes and a yeast, Aureobasidium pullulans. The inactivation ratio was experimentally measured as a function of sonication time for different ultrasound power and for different initial cell numbers. For both E. aerogenes and A. pullulans the Log of the inactivation ratio decreased linearly with sonication time, and the rate of inactivation increased (D-value decreased) with the increase in sonication power. The rate of inactivation was also found, for both microorganisms, to increase with a decrease in the initial cell number. The fits, obtained using the proposed model, are in very good agreement with the experimental data.     

Effect of ultrasound treatment on steady and dynamic shear properties of glucomannan based salep dispersions: optimization of amplitude level, sonication time and temperature using response surface methodology

The present study investigated effect of different amplitude levels (40, 70 and 100%), sonication temperatures (40, 50 and 60 °C) and exposure times (3, 7 and 11 min) on steady shear properties; apparent viscosity (η), shear stress (σ), consistency coefficient (K), flow behavior index (n) and dynamic shear properties; storage modulus (G′), loss modulus (G″), complex viscosity (η^∗), complex modulus (G^∗) and loss tangent (tan δ) values of glucomannan based salep solution (SS) and salep drink (SD) samples. In addition, the steady and dynamic shear properties were optimized using ridge analysis in terms of amplitude level, sonication temperature and exposure times levels. Increasing amplitude level and sonication time decreased considerably the η, σ, K, G′, G″ and η^∗ values of salep dispersions (SS and SD samples). However, sonication temperature did not have a remarkable effect on these properties.     

Ultrasound induces cellular destruction of nasopharyngeal carcinoma cells in the presence of curcumin

Objectives

Curcumin, a natural pigment from the traditional Chinese herb, has shown promise as an efficient enhancer of ultrasound. The present study aims to investigate ultrasound-induced cellular destruction of nasopharyngeal carcinoma cells in the presence of curcumin in vitro.

Methods

Nasopharyngeal carcinoma cell line CNE2 cells were incubated by 10 μm curcumin and then were treated by ultrasound for 8 s at the intensity of 0.46 W/cm². Cytotoxicity was evaluated using MTT assay and light microscopy. Mitochondrial damage was analyzed using a confocal laser scanning microcopy with Rhodamine 123 and ultrastructural changes were observed using a transmission electron microscopy (TEM).

Results

MTT assay showed that cytotoxicity induced by ultrasound treatment alone and curcumin treatment alone was 18.16 ± 2.37% and 24.93 ± 8.30%, respectively. The cytotoxicity induced by the combined treatment of ultrasound and curcumin significantly increased up to 86.67 ± 7.78%. TEM showed that microvillin disappearance, membrane blebbing, chromatin condensation, swollen mitochondria, and mitochondrial myelin-like body were observed in the cells treated by ultrasound and curcumin together. The significant collapse of mitochondrial membrane potential (MMP) was markedly observed in the CNE2 cells after the combined treatment of curcumin and ultrasound.

Conclusions

Our findings demonstrated that ultrasound sonication in the presence of curcumin significantly killed the CNE2 cells and induced ultrastructural damage and the dysfunction of mitochondria, suggesting that ultrasound treatment remarkably induced cellular destruction of nasopharyngeal carcinoma cells in the presence of curcumin.     

Effects of low-intensity ultrasound on the growth,cell membrane permeability and ethanol tolerance of Saccharomyces cerevisiae

Effects of low-intensity ultrasound (at different frequency, treatment time and power) on Saccharomyces cerevisiae in different growth phase were evaluated by the biomass in the paper. In addition, the cell membrane permeability and ethanol tolerance of sonicated Saccharomyces cerevisiae were also researched. The results revealed that the biomass of Saccharomyces cerevisiae increased by 127.03% under the optimum ultrasonic conditions such as frequency 28 kHz, power 140 W/L and ultrasonic time 1 h when Saccharomyces cerevisiae cultured to the latent anaphase. And the membrane permeability of Saccharomyces cerevisiae in latent anaphase enhanced by ultrasound, resulting in the augment of extracellular protein, nucleic acid and fructose-1,6-diphosphate (FDP) contents. In addition, sonication could accelerate the damage of high concentration alcohol to Saccharomyces cerevisiae although the ethanol tolerance of Saccharomyces cerevisiae was not affected significantly by ultrasound.     

Invert sugar formation with Saccharomyces cerevisiae cells encapsulated in magnetically responsive alginate microparticles

Invert sugar (an equimolar mixture of glucose and fructose prepared by sucrose hydrolysis) is a very important food component. We have prepared magnetically responsive alginate microbeads containing entrapped Saccharomyces cerevisiae cells and magnetite microparticles which can be easily separated in an appropriate magnetic separator. The microbeads (typical diameter between 50 and 100 μm) were prepared using the water-in-oil emulsification process. The prepared microbeads containing yeast cells with invertase activity enabled efficient sucrose conversion. The biocatalyst was quite stable; the same catalytic activity was observed after one month storage at 4 °C and the microbeads could be used at least six times.     

Third-order nonlinear optical properties of thin sputtered gold films

Au films of thickness ranging between 5 and 52 nm were prepared by sputtering on quartz substrates and their third-order nonlinear optical response was investigated by Optical Kerr effect (OKE) and Z-scan techniques using 532 nm, 35 ps laser pulses. All prepared films were characterized by XRD, AFM and UV-VIS-NIR spectrophotometry while their third-order susceptibility χ⁽³⁾ was measured and found to be of the order of 10⁻⁹ esu. The real and imaginary parts of the third-order susceptibility were found in very good agreement with experimental results and theoretical predictions reported by Smith et al. [D.D. Smith, Y. Yoon, R.W. Boyd, Y.K. Cambell, L.A. Baker, R.M. Crooks, M. George, J. Appl. Phys. 86 (1999) 6200].