Yacine Hemar

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.

Quantification of high-power ultrasound induced damage on potato starch granules using light microscopy

A simple light microscopic technique was developed in order to quantify the damage inflicted by high-power low-frequency ultrasound (0-160 W, 20 kHz) treatment on potato starch granules in aqueous dispersions. The surface properties of the starch granules were modified using ethanol and SDS washing methods, which are known to displace proteins and lipids from the surface of the starch granules. The study showed that in the case of normal and ethanol-washed potato starch dispersions, two linear regions were observed. The number of defects first increased linearly with an increase in ultrasound power up to a threshold level. This was then followed by another linear dependence of the number of defects on the ultrasound power. The power threshold where the change-over occurred was higher for the ethanol-washed potato dispersions compared to non-washed potato dispersions. In the case of SDS-washed potato starch, although the increase in defects was linear with the ultrasound power, the power threshold for a second linear region was not observed. These results are discussed in terms of the different possible mechanisms of cavitation induced-damage (hydrodynamic shear stresses and micro-jetting) and by taking into account the hydrophobicity of the starch granule surface.

Non-targeted analysis of tea by hydrophilic interaction liquid chromatography and high resolution mass spectrometry.

Tea is the second most consumed beverage in the world and its consumption has been associated with numerous potential health benefits. Factors such as fermentation methods, geographical origin and season can affect the primary and secondary metabolite composition of tea. In this study, a hydrophilic interaction liquid chromatography (HILIC) method coupled to high resolution mass spectrometry in both positive and negative ionisation modes was developed and optimised. The method when combined with principal component analysis to analyse three different types of tea, successfully distinguished samples into different categories, and provided evidence of the metabolites which differed between them. The accurate mass and high resolution attributes of the mass spectrometric data were utilised and relative quantification data were extracted post-data acquisition on 18 amino acids, showing significant differences in amino acid concentrations between tea types and countries. This study highlights the potential of HILIC chromatography combined with non-targeted mass spectrometric methods to provide a comprehensive understanding of polar metabolites in plant extracts.

Monitoring tea fermentation/manufacturing by direct analysis in real time (DART) mass spectrometry

Factors such as fermentation methods, geographical origin and season can affect the biochemical composition of tea leaves (Camellia sinensis L.). In this study, the biochemical composition of oolong tea during the manufacturing and fermentation process was studied using a non-targeted method utilising ambient ionisation with a direct analysis in real time (DART) ion source and mass spectrometry (MS). Caffeine dominated the positive ionisation spectra throughout the manufacturing process, while the negative ion spectra collected during manufacturing were rich in ions likely to be surface lipids. Correlation analyses on the spectra revealed two volatile compounds tentatively identified as indole and geranic acid, along with ammonium and caffeine clusters/adducts with geranic acid that increased in concentration during the fermentation stages of the process. The tentative identifications were assigned using a combination of DART-ion-trap MS(n) and DART-accurate mass MS(1) and MS(2) on tea samples and standard compounds. This study highlights the potential of DART-MS to rapidly monitor the progress of complex manufacturing processes such as tea fermentation.

Nonlinear Behavior of Gelatin Networks Reveals a Hierarchical Structure

We investigate the strain hardening behavior of various gelatin networks-namely physical gelatin gel, chemically cross-linked gelatin gel, and a hybrid gel made of a combination of the former two-under large shear deformations using the pre-stress, strain ramp, and large amplitude oscillations shear protocols. Further, the internal structures of physical gelatin gels and chemically cross-linked gelatin gels were characterized by small angle neutron scattering (SANS) to enable their internal structures to be correlated with their nonlinear rheology. The Kratky plots of SANS data demonstrate the presence of small cross-linked aggregates within the chemically cross-linked network whereas, in the physical gelatin gels, a relatively homogeneous structure is observed. Through model fitting to the scattering data, we were able to obtain structural parameters, such as the correlation length (ξ), the cross-sectional polymer chain radius (R(c)) and the fractal dimension (d(f)) of the gel networks. The fractal dimension d(f) obtained from the SANS data of the physical and chemically cross-linked gels is 1.31 and 1.53, respectively. These values are in excellent agreement with the ones obtained from a generalized nonlinear elastic theory that has been used to fit the stress-strain curves. The chemical cross-linking that generates coils and aggregates hinders the free stretching of the triple helix bundles in the physical gels.

The effect of ultrasound on particle size, color, viscosity and polyphenol oxidase activity of diluted avocado puree

The effect of ultrasound treatment on particle size, color, viscosity, polyphenol oxidase (PPO) activity and microstructure in diluted avocado puree was investigated. The treatments were carried out at 20 kHz (375 W/cm(2)) for 0-10 min. The surface mean diameter (D[3,2]) was reduced to 13.44 μm from an original value of 52.31 μm by ultrasound after 1 min. A higher L(∗) value, ΔE value and lower a(∗) value was observed in ultrasound treated samples. The avocado puree dilution followed pseudoplastic flow behavior, and the viscosity of diluted avocado puree (at 100 s(-1)) after ultrasound treatment for 1 min was 6.0 and 74.4 times higher than the control samples for dilution levels of 1:2 and 1:9, respectively. PPO activity greatly increased under all treatment conditions. A maximum increase of 25.1%, 36.9% and 187.8% in PPO activity was found in samples with dilution ratios of 1:2, 1:5 and 1:9, respectively. The increase in viscosity and measured PPO activity might be related to the decrease in particle size. The microscopy images further confirmed that ultrasound treatment induced disruption of avocado puree structure.

Conjugation of Bovine Serum Albumin and Glucose under Combined High Pressure and Heat

The effect of combined heat and pressure on the Maillard reaction between bovine serum albumin (BSA) and glucose was investigated. The effects in the range of 60-132 °C and at 0.1-600 MPa on the lysine availability of BSA were investigated at isothermal/isobaric conditions. The kinetic results showed that the protein-sugar conjugation rate increased with increasing temperature, whereas it decreased with increasing pressure. The reaction followed 1.4th order kinetics at most conditions investigated. A mathematical model describing BSA-glucose conjugation kinetics as a function of pressure and temperature is proposed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry were used to verify BSA-glucose conjugation and to identify the glucosylated sites. These indicated that the application of combined high pressure and high temperature resulted in significant differences in the progression of the Maillard reaction as compared to heat treatments at atmospheric pressure.

In situ study of maize starch gelatinization under ultra-high hydrostatic pressure using X-ray diffraction

The gelatinization of waxy (very low amylose) and high-amylose maize starches by ultra-high hydrostatic pressure (up to 6 GPa) was investigated in situ using synchrotron X-ray powder diffraction on samples held in a diamond anvil cell (DAC). The starch pastes, made by mixing starch and water in a 1:1 ratio, were pressurized and measured at room temperature. X-ray diffraction pattern showed that at 2.7 GPa waxy starch, which displayed A-type XRD pattern at atmospheric pressure, exhibited a faint B-type-like pattern. The B-type crystalline structures of high-amylose starch were not affected even when 1.5 GPa pressure was applied. However, both waxy and high-amylose maize starches can be fully gelatinized at 5.9 GPa and 5.1 GPa, respectively. In the case of waxy maize starch, upon release of pressure (to atmospheric pressure) crystalline structure appeared as a result of amylopectin aggregation.

The physicochemical properties of a new class of anticancer fungal polysaccharides: a comparative study.

The structural and physicochemical properties of polysaccharides isolated from fungi with anticancer properties were investigated. The majority of the polysaccharides considered, have the β-d-Glcp component mostly connected by 1→3 and 1→6 linkages in the backbones and the short branches, respectively. The established parameters of lead-like, drug-like and of known dug space (KDS) were used and the repeating units of the polysaccharides exhibit some overlap with these. It was found that a unique region of chemical space is occupied by the polysaccharides, with MW: 1.0 x 10(5) to 2.5 x 10(5) g mol(-1); LogP: -3.0 x 10(3) to -1.0 x 10(3); HD: 1.0 x 10(3) to 5.0 x 10(3); HA: 5.0 x 10(3) to 1.0 x 10(4); PSA: 5.0 x 10(4) to 1.0 x 10(5) and RB: 5.0 x 10(3) to 1.0 x 10(4). These findings can be exploited in antitumor drug discovery projects.

Inactivation of Enterobacter aerogenes in reconstituted skim milk by high- and low-frequency ultrasound

The inactivation of Enterobacter aerogenes in skim milk using low-frequency (20kHz) and high-frequency (850kHz) ultrasonication was investigated. It was found that low-frequency acoustic cavitation resulted in lethal damage to E. aerogenes. The bacteria were more sensitive to ultrasound in water than in reconstituted skim milk having different protein concentrations. However, high-frequency ultrasound was not able to inactivate E. aerogenes in milk even when powers as high as 50W for 60min were used. This study also showed that high-frequency ultrasonication had no influence on the viscosity and particle size of skim milk, whereas low-frequency ultrasonication resulted in the decrease in viscosity and particle size of milk. The decrease in particle size is believed to be due to the breakup of the fat globules, and possibly to the cleavage of the κ-casein present at the surface of the casein micelles. Whey proteins were also found to be slightly affected by low-frequency ultrasound, with the amounts of α-lactalbumin and β-lactoglobulin slightly decreasing.