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Biomedical subjects

P Gervais

Publications and source records attributed to P Gervais.

At least 19 recordsLinked to original sources

Water activity affects heat resistance of microorganisms in food powders.

To study the factors and mechanisms involved in microorganisms' death or resistance to temperature in low-water-activity environments, a previous work dealt with the viability of dried microorganisms immobilized in thin-layer on glass beads. This work is intended to check the efficiency of a rapid heating-cooling treatment to destroy microorganisms that were dried after mixing with wheat flour or skim milk. The thermoresistance of the yeast Saccharomyces cerevisiae and the bacterium Lactobacillus plantarum were studied. Heat stress was applied at two temperatures (150 or 200 degrees C) for treatments of one of four durations (5, 10, 20, or 30 s) and at seven levels of initial water activity (a(w)) in the range 0.10 to 0.70. This new treatment achieved a microbial destruction of eight log reductions. A specific initial water activity was defined for each strain at which it was most resistant to heat treatments. On wheat flour, this initial a(w) value was in the range 0.30-0.50, with maximal viability value at a(w)=0.35 for L. plantarum, whatever the temperature studied, and 0.40 for S. cerevisiae. For skim milk, a variation in microbial viability was observed, with optimal resistance in the range 0.30-0.50 for S. cerevisiae and 0.20-0.50 for L. plantarum, with minimal destruction at a(w)=0.30 whatever the heating temperature is.

Food Contamination↗

Pressure-induced shape change of phospholipid vesicles: implication of compression and phase transition.

A microscopic study has allowed the analysis of modifications of various shapes acquired by phospholipid vesicles during a hydrostatic pressure treatment of up to 300 MPa. Giant vesicles of dimyristoylphosphatidylcholine / phosphatidylserine (DMPC/PS) prepared at 40 degrees C mainly presented a shape change resembling budding during pressure release. This comportment was reinforced by the incorporation of 1,2-dioleyl-sn-glycero-3-phosphatidylethanolamine (DOPE) or by higher temperature (60 degrees C) processing. The thermotropic main phase transition (L alpha to P beta') of the different vesicles prepared was determined under pressure through a spectrofluorimetric study of 6-dodecanoyl-2-dimethylamino-naphtalene (Laurdan) incorporated into the vesicles' bilayer. This analysis was performed by microfluorescence observation of single vesicles. The phase transition was found to begin at about 80 MPa and 120 MPa for DMPC/PS vesicles at, respectively, 40 degrees C and 60 degrees C. At 60 degrees C the liquid-to-gel transition phase was not complete within 250 MPa. Addition of DMPE at 40 degrees C does not significantly shift the onset boundary of the phase transition but extends the transition region. At 40 degrees C, the gel phase was obtained at, respectively, 110 MPa and 160 MPa for DMPC/PS and DMPC/PS/DOPE vesicles. In comparing volume data obtained from image analysis and Laurdan signal, we assume the shape change is a consequence of the difference between lateral compressibility of the membrane and bulk water. The phase transition contributes to the membrane compression but seems not necessary to induce shape change of vesicles. The high compressibility of the L alpha phase at 60 degrees C allows induction on DMPC/PS vesicles of a morphological transition without phase change.

Compressive Strength↗

Air drying optimization of Saccharomyces cerevisiae through its water-glycerol dehydration properties.

AIMS: This study describes the different stages of optimization in an original drying process for yeasts, which allows the retrieval of dried samples of Saccharomyces cerevisiae CBS 1171 with maximum viability. METHODS AND RESULTS: The process involves the addition of wheat flour to yeast pellets, followed by mixing and then air-drying in a fluidized bed dryer. The sensitivity to the osmotic stress was first studied in a water-glycerol solution and the observed results were then applied to the drying process. This study have shown that the yeast was quite resistant to osmotic stress and pointed out the existence of zones of sensitivity where viability dramatically decrease as function of final osmotic pressure and temperature of the treatment. Thus, for dehydration until low osmotic pressure (133 MPa, i.e. a(w) = 0.38) results have shown that viability was better when temperature of the treatment was less than 8 degrees C or higher than 25 degrees C. Moreover, kinetic of dehydration was found to greatly influence cells recovery. CONCLUSIONS: These observations allowed the choice of parameters of dehydration of yeasts with an original drying process which involve the mix of the yeasts with wheat flour and then drying in a fluidized bed. SIGNIFICANCE AND IMPACT OF THE STUDY: This process dried rapidly the yeasts to less than 220 MPa (aw < or = 0.2) with whole cell recovery and good fermentative capabilities.

Cell Survival↗

Thermal properties and granulometry of dried powders strongly influence the effectiveness of heat treatment for microbial destruction.

The thermal treatment of Saccharomyces cerevisiae cells, which were homogeneously incorporated into dried wheat flour particles, was performed for various particle radii (0.8 to 1.6, 1.6 to 2.8, 2.8 to 3.2, and 5 mm) and for an initial water activity of 0.20. A new high-temperature short-time process developed by our laboratory for powder decontamination was used at 150, 200, and 250 degrees C for 5 to 30 s, and significant destruction of up to a 6.7-log reduction, depending on treatment conditions and granule size, was achieved. This study confirms the strong influence of granulometry on the microbial destruction of homogeneously contaminated powdered products. Moreover, a thermal model was developed that takes into account the thermal properties of each component, the variations during heat treatment, and the energy required for phase change. This model provides a tool for predicting yeast destruction.

Colony Count, Microbial↗

Death of Escherichia coli during rapid and severe dehydration is related to lipid phase transition.

This study reports the effects of exposure to increasing osmotic pressure on the viability and membrane structure of Escherichia coli. Changes in membrane structure after osmotic stress were investigated by electron transmission microscopy, measurement of the anisotropy of the membrane fluorescent probe DPH (1,6-diphenyl-1,3,5-hexatriene) inserted in E. coli, and Fourier infrared spectroscopy (FTIR). The results show that, above a critical osmotic pressure of 35 MPa, the viability of the bacterium is drastically reduced (2 log decrease in survivors). Electron micrographs revealed a severe contraction of the cytoplasm and the formation of membrane vesicles at 40 MPa. Changes in DPH anisotropy showed that osmotic dehydration to 40 MPa promoted a decrease in the membrane fluidity of integral cells of E. coli. FTIR measurements showed that at 10-40 MPa a transition from lamellar liquid crystal to lamellar gel among the phospholipids extracted from E. coli occurred. Bacterial death resulting from dehydration can be attributed to the conjunction between membrane deformation, caused by the volumetric contraction, and structural changes of the membrane lipids. The influence of the latter on the formation of membrane vesicles and on membrane permeabilization at lethal osmotic pressure is discussed, since vesiculation is hypothetically responsible for cell death.

Cell Membrane↗

Efficiency of pulsed UV light for microbial decontamination of food powders.

The aim of this study was to evaluate the efficiency of pulsed light on the destruction of dried microorganisms on fluidized glass beads and to determine treatment parameters (energy level, water activity, final product quality) for process optimization. The applied drying method allowed microorganisms to remain viable on glass beads or dried powdered products with viability yields approaching 100%. The pulsed UV light system enabled an efficient fluidization of food powders, even for granular products (up to 5 mm diameter) and avoided shadowed areas. For Saccharomyces cerevisiae decontamination, the dose effect of UV rays was preponderant with glass beads and quartz plate, and in this case, 58 J/cm2 were required to decrease the microbial population by 7 log. For colored food powders (black pepper and wheat flour), the thermal effect of pulsed light dominated the UV effect.

Consumer Product Safety↗

Achievement of rapid osmotic dehydration at specific temperatures could maintain high Saccharomyces cerevisiae viability.

Various methods have been tried to prevent cell mortality during dehydration, but the reasons why microorganisms die when submitted to dehydration and rehydration are not well understood. The aim of this study was to further investigate the reasons for yeast mortality during dehydration. Osmotic dehydration and rehydration of Saccharomyces cerevisiae W303-1A were performed at different temperatures. Two different approaches were used: isothermic treatments (dehydration and rehydration at the same temperature), and cyclic treatments (dehydration at an experimental temperature and rehydration at 25 degrees C), with significant differences in viability found between the different treatments. Dehydration at lower and higher temperatures gave higher viability results. These experiments allowed us to propose a hypothesis that relates mortality to a high water flow through an unstable membrane during phase transition.

Dehydration↗

Unexpected thermal destruction of dried, glass bead-immobilized microorganisms as a function of water activity.

To help us understand the factors and mechanisms implicated in the death of microorganisms or their resistance to temperature in a low water activity environment, microorganisms were dried on the surface of glass beads before being subjected to high temperatures for a short period followed by rapid cooling. Two microorganisms were studied: the yeast Saccharomyces cerevisiae and the bacterium Lactobacillus plantarum. Experiments were carried out at 150, 200, and 250 degrees C, with four durations of heat treatment and seven levels of initial water activity between 0.10 and 0.70. We observed an unexpected range of water activity, between 0.30 and 0.50, at which microorganisms were more resistant to the various treatments, with maximal viability at 0.35 for L. plantarum and 0.40 for S. cerevisiae.

Food Microbiology↗

Involvement of osmotic cell shrinkage on the proton extrusion rate in Saccharomyces cerevisiae.

Saccharomyces cerevisiae has been subjected to hyperosmotic shocks by using permeating (sorbitol, xylitol, glycerol, NaCl) and nonpermeating (PEG 600) solutes. The proton extrusion rate decreased as the osmotic pressure increased, whichever solute was used. However, the total inhibition of the cellular H+ extrusion depended on the solute used. A total inhibition was observed at about 20 MPa with glycerol, xylitol and sorbitol. With PEG 600, a total inhibition of extracellular acidification was obtained at 8.5 MPa. NaCl, with an extracellular pressure of 37.8 MPa (near saturation), did not completely inhibit the extracellular acidification. These results showed that the total inhibition of proton extrusion, involving probably the membrane H+-ATPase. was not correlated to the hydric state of the external medium but was strictly linked to the degree of permeation of solutes across the plasma membrane. The extracellular acidification was totally inhibited by a critical final cell volume reached after the osmotic shrinkage, whichever solute was used. This critical final cell volume represented 50% of the initial cell volume. This result suggests that the final cell volume reached after an osmotic stress represents a key thermodynamic parameter for cell osmoregulation in which H+-ATPase would be implicated.

Cell Membrane↗

Upstream use of tirofiban in patients admitted for an acute coronary syndrome in hospitals with or without facilities for invasive management. PRISM-PLUS Investigators.

Management and prognosis of acute coronary syndromes may be influenced by the availability of catheterization facilities at admitting hospitals. Treatment effects of tirofiban were examined in a Canadian cohort of 834 patients enrolled in the Canadian Platelet Receptor Inhibition in Ischemic Syndrome Management in Patients Limited by Unstable Signs and Symptoms (PRISM-PLUS) trial according to admission into hospitals without (n = 322) or with catheterization facilities (n = 512). Hospital transfers for cardiac catheterization were facilitated using preexisting networks accelerated for the purposes of the protocol. In hospitals without facilities, the relative risks for occurrence of death, infarction, or refractory ischemia among patients receiving tirofiban plus heparin compared with heparin alone were 0.52 at 7 days (p = 0.02), 0.59 at 30 days (p = 0.03), and 0.70 at 180 days (p = 0.09); and for death or infarction, 0.32 (p = 0.02), 0.46 (p = 0.04,) and 0.51 (p = 0.03), respectively. Benefit was seen regardless of transfer status, with no statistically significant interaction between treatment, hospital type, and catheterization for any end point at any time point. The incidence of Thrombolysis In Infarction defined major bleeding with respect to therapy was not significantly different between hospital types. Thus, upstream treatment with tirofiban plus heparin confers clinical benefits in unstable angina and/or non-ST-segment elevation infarction patients regardless of whether initial presentation is to a hospital without catheterization facilities or to a hospital with such facilities.

Acute Disease↗

The effect of osmotic pressure on the membrane fluidity of Saccharomyces cerevisiae at different physiological temperatures.

Membrane fluidity in whole cells of Saccharomyces cerevisiae W303-1A was estimated from fluorescence polarization measurements using the membrane probe, 1,6-diphenyl-1,3,5-hexatriene, over a wide range of temperatures (6-35 degrees C) and at seven levels of osmotic pressure between 1.38 MPa and 133.1 MPa. An increase in phase transition temperatures was observed with increasing osmotic pressure. At 1.38 MPa, a phase transition temperature of 12 +/- 2 degrees C was observed, which increased to 17 +/- 4 degrees C at 43.7 MPa, 21+/- 7 degrees C at 61.8 MPa, and 24 +/- 9 degrees C at an osmotic pressure of 133.1 MPa. From these results we infer that, with increases in osmotic pressure, the change in phospholipid conformation occurs over a larger temperature range. These results allow the representation of membrane fluidity as a function of temperature and osmotic pressure. Osmotic shocks were applied at two levels of osmotic pressure and at nine temperatures, in order to relate membrane conformation to cell viability.

Membrane Fluidity↗

Coupling effects of osmotic pressure and temperature on the viability of Saccharomyces cerevisiae.

The osmotic tolerance of cells of Saccharomyces cerevisiae as a function of glycerol concentration and temperature has been investigated. Results show that under isothermal conditions (25 degrees C) cells are resistant (94% viability) to hyperosmotic treatment at 49.2 MPa. A thigher osmotic pressure, cell viability decreases to 25% at 99 MPa. Yeast resistance to high osmotic stress (99 Mpa) is enhanced at low temperatures (5-11 degrees C). Therefore, the temperature at which hyperosmotic pressure is achieved greatly affects cell viability. These results suggest that temperature control is a suitable means of enhancing cell survival in response to osmotic dehydration.

Cold Temperature↗

Influence of the fluidity of the membrane on the response of microorganisms to environmental stresses.

The aim of this mini-review is to relate membrane physical properties to the adaptation and resistance of microorganisms to environmental stresses. In the first part, the effects of various stresses on the structure and dynamic properties of phospholipid and biological membranes are presented. The compensation of these effects, i.e., change in membrane fluidity, phase transitions, by the active cellular control of the membrane chemical composition, is then described. In this natural process, the change in membrane fluidity is viewed as the detecting "input" signal that initiates the regulation, activating proteic effectors that in turn may influence the chemical composition of the membrane (feedback). This adaptation system allows the maintenance of the physical characteristics of membranes and, thereby, of their functionality. When environmental stresses are extreme and occur abruptly, the regulation process may not compensate for the changes in the membrane physical characteristics. In such cases, important variations in the membrane fluidity and structure may induce cellular damages and cell death. However, the lethal consequences are not systematically observed because protective effects of changes in the membrane physical state on the resistance to stresses are also reported.

Membrane Fluidity↗

Pressure-temperature phase transition diagram for wheat starch.

Wheat starch suspensions in water (5% dry matter) were subjected to various pressures (0.1-600 MPa) and temperatures (-20 to 96 degrees C) for 15 min. The gelatinization rate was measured after treatment by using microscopic measurements of the loss of birefringence of the granules. This method was previously calibrated by differential scanning calorimetry. Curves of isogelatinization were found to be quite similar to a pressure-temperature (P-T) diagram of unfolding proteins. Results were first analyzed by considering the thermodynamic aspects related to the dT/dP curve shifts. On the basis of equations already shown for proteins, the P-T gelatinization diagram of wheat starch would show different kinds of thermal contributions, suggesting endothermic, athermic, or exothermic melting reactions. Second, as a practical consequence, these previous P-T areas corresponded to specific gelatinization conditions as confirmed by hydration evaluation measured by starch swelling index. Depending on the pressure-temperature conditions, gelatinization would involve hydration. Lowering the pressure and temperature resulted in a complete gelatinization with less hydration in comparison with a thermal treatment at atmospheric pressure. A hydration model based on an energetic approach was proposed.

Birefringence↗

Osmotic mass transfer in the yeast Saccharomyces cerevisiae.

This paper reviews the passive mechanisms involved in the response of a yeast to changes in medium concentration and osmotic pressure. The results presented here were collected in our laboratory during the last decade and are experimentally based on the measurement of cell volume variations in response to changes in the medium composition. In the presence of isoosmotic concentration gradients of solutes between intracellular and extracellular media, mass transfers were found to be governed by the diffusion rate of the solutes through the cell membrane and were achieved within a few seconds. In the presence of osmotic gradients, mass transfers mainly consisting in a water flow were found to be rate limited by the mixing systems used to generate a change in the medium osmotic pressure. The use of ultra-rapid mixing systems allowed us to show that yeast cells respond to osmotic upshifts within a few milliseconds and to determine a very high hydraulic permeability for yeast membrane (Lp>6.10(-11) m x sec)-1) x Pa(-1)). This value suggested that yeast membrane may contain facilitators for water transfers between intra and extracellular media, i.e. aquaporins. Cell volume variation in response to osmotic gradients was only observed for osmotic gradients that exceeded the cell turgor pressure and the maximum cell volume decrease, observed during an hyperosmotic stress, corresponded to 60% of the initial yeast volume. These results showed that yeast membrane is highly permeable to water and that an important fraction of the intracellular content was rapidly transferred between intracellular and extracellular media in order to restore water balance after hyperosmotic stresses. Mechanisms implied in cell death resulting from these stresses are then discussed.

Animals↗

Influence of thermal and osmotic stresses on the viability of the yeast Saccharomyces cerevisiae.

This work studies the effect of thermal and dehydration kinetics on the viability of Saccharomyces cerevisiae. The influence of the rate of temperature (T) and osmotic pressure (pi) increases are first investigated. Results showed that yeast viability is preserved by slow variations of temperature or osmotic pressure in a precise range of T or pi. The influence of a previous thermal stress on the resistance to a hyperosmotic stress is also studied. Temperatures equal to or lower than 10 degrees C allowed the preservation of viability after an osmotic stress whereas temperatures above 10 degrees C did not preserve yeast survival.

Hot Temperature↗

Volume changes of isolated human K562 leukemia cells induced by electric field pulses.

Electropermeabilization of immobilized human leukemia K562 cells was studied by measuring changes in cell volume. Such changes reflect mass transfer between the cell and external medium. Electropermeabilization was carried out in an isosmotic water-sorbitol medium with a range of electric field strengths from 500 to 800 V. cm(-1), corresponding to low-energy levels. Electroporation of the K562 cell membrane was found to provoke an inflow of sorbitol and a corresponding osmotic inflow of water and/or an outflow of intracellular solutes due to Fick diffusion. Such flows were found to involve the shrinkage, swelling, or rupture of K562 cells, depending on the characteristics of the electric field and of the physiological state of cells. The behavior of immobilized cells was observed during their exposure to the electric field. The response in immobilized cell volume corresponded with the theoretical pore size and pore opening time, permitting an explanation of the behavior of cell suspensions subject to electrical fields.

Cell Size↗

Amyloglucosidase hydrolysis of high-pressure and thermally gelatinized corn and wheat starches.

The study of glucose production using amyloglucosidase as a biocatalyst was carried out using high-pressure and thermally gelatinized corn and wheat starches. For corn starch, the measured initial rate of glucose production obtained from thermal gelatinization is faster than that obtained from the two high-pressure treatments, but the equilibrium yield of glucose was found to be similar for the three treatments. High-pressure treatments of wheat starch significantly improve the equilibrium yield of glucose compared with those obtained from the thermally gelatinized wheat starch. This difference has been related to the formation of amylose-lipid complexes during heating and could also explain previous physicochemical differences observed between high-pressure and thermally gelatinized starch.

Catalysis↗