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At least 325 records · Page 18Linked to original sources

PubMed1977

Initiation of Bacillus spore germination by hydrostatic pressure: effect of temperature.

Suspensions of Bacillus cereus T, B. subtilis, and B. pumilus spores in water or potassium phosphate buffer were germinated by hydrostatic pressures of between 325 and 975 atm. Kinetics of germination at temperatures within the range of 25 to 44 degrees C were determined, and thermodynamic parameters were calculated. The optimum temperature for germination was dependent on pressure, species, suspending medium, and storage time after heat activation. Germination rates increased significantly with small increments of pressure, as indicated by high negative deltaV values of -230 +/- 5 cm3/mol for buffered B. subtilis (500 to 700 atm) and B. pumilus (500 atm) spores and -254 +/- 18 cm3/mol for aqueous B. subtilis (400 to 550 atm) spores at 40 degrees C and -612 +/- 41 cm3/mol for B. cereus (500 to 700 atm) spores at 25 degrees C. The ranges of thermodynamic constants calculated at 40 degrees C for buffered B. pumilus and B. subtilis spores at 500 and 600 atm and for aqueous B. subtilis spores at 500 atm were: Ea = 181,000 to 267,000 J/mol; deltaH = 178,000 to 264,000 J/mol; deltaG = 94,000 to 98,300 J/mol; deltaS = 264 to 544 J/mol per degree K. These values are consistent with the concept that the transformation of a dormant to a germinating spore induced by hydrostatic pressure involves either hydration or a reduction in the visocosity of the spore core and a conformational change of an enzyme.

Bacillus↗

PubMed2002

Dissecting the molecular origins of specific protein-nucleic acid recognition: hydrostatic pressure and molecular dynamics.

The fundamental processes by which proteins recognize and bind to nucleic acids are critical to understanding cellular function. To explore the factors involved in protein-DNA recognition, we used hydrostatic pressure to perturb the binding of the BamHI endonuclease to cognate DNA, both in experiment and in molecular dynamic simulations. A new technique of high-pressure gel mobility shift analysis was used to test the effects of elevated hydrostatic pressure on the binding of BamHI to its cognate recognition sequence. Upon application of a pressure of 500 bar, the equilibrium dissociation constant of BamHI binding to the cognate site was found to increase nearly 10-fold. A challenge has been to link this type of pure thermodynamic measurement to functional events occurring at the molecular level. Thus, we used molecular dynamic simulations at both ambient and elevated pressures to reveal details of the direct and water-mediated interactions between BamHI and cognate DNA, which allow explanation of the effects of pressure on site-specific protein-DNA binding and complex stability.

Base Sequence↗

PubMed1983

Anesthetic antagonism of the effects of high hydrostatic pressure on locomotory activity of the brine shrimp Artemia.

The locomotory activity of small groups of brine shrimp (Artemia salina) was studied under conditions of high hydrostatic pressure, varying temperatures and exposure to several gaseous anesthetics. Both compression and exposure to anesthesia reduced the animal's swimming activity, while temperature increased or decreased activity as it was raised or lowered from ambient. The effect of the anesthetics was less during periods of simultaneous exposure to high hydrostatic pressure. It is concluded that pressure antagonism of anesthesia is demonstrable in invertebrate organisms and may represent a fundamental interaction of these parameters in biological systems.

Anesthetics↗

PubMed1991

Vesiculation induced by hydrostatic pressure in human erythrocytes.

When human erythrocytes were subjected to hydrostatic pressure (1.1-2.0 kbar), it was found that membrane vesicles were released from the red cells above 1.4 kbar. As with hemolysis under high pressure, the amount of released vesicles was increased with increasing pressure but decreased by the cross-linking of membrane proteins with diamide. Vesicles obtained at 2.0 kbar were heterogeneous in size but similar to intact erythrocytes in phospholipid composition. Although it has been reported that spectrin-free vesicles are released by echinocytogenic agents, pressure-induced vesicles did contain considerable and similar amounts of spectrin irrespective of the difference in size. These results suggest that vesiculation by high pressure is associated with the disruption of the membrane skeleton, as previously seen in pressure-induced hemolysis [Yamaguchi et al. (1989) J. Biochem. 106, 1080-1085].

Adult↗

PubMed1989

[Hydrostatic pressure in the duodenum during the immediate postoperative period after resection of the stomach].

Twenty-nine patients were examined after resection of the stomach by various methods. The hydrostatic pressure value was found to be independent on the method of resection. It was maximum on the third postoperative day in all patients. No correlation was established between the hydrostatic pressure and the amount of duodenal contents discharged along the nasoduodenal catheter.

Duodenum↗

PubMed1979

Dissociation and aggregation of lactic dehydrogenase by high hydrostatic pressure.

As shown by earlier experiments high hydrostatic pressure affects the catalytic function of lactic dehydrogenase from rabbit muscle. In the presence of substrates denaturation occurs, whereas in the absence of substrates and --SH-protecting reagents oxidation of sulfhydryl groups takes place [Schmid, G., Lüdemann, H.-D. & Jaenicke, R. (1975) Biophys. Chem. 3, 90--98; (1978) Eur. J. Biochem. 86, 219--224]. Avoiding oxidation effects by reducing conditions in the solvent medium and by chelation of heavy metal ions, the remaining high-pressure effects consist of dissociation of the native quaternary structure into subunits followed by aggregation. Both reactions are influenced by temperature and enzyme concentration. Short incubation (less than or equal to 10 min) at pH 6.0--8.5 and pressures of 0.3--1.0 kbar causes dissociation which is reversed at normal pressure. At 5 degrees C the activation volume is found to be delta V not equal to = -62 +/- 3cm3 . mol-1. Above 1.2 kbar irreversible aggregation takes place; the reaction is favoured by low temperature and decreased pH. The activation volume for the aggregation process at 5 degress C is delta V not equal to = -97 +/- 3cm3 . mol-1. The results may be described by a reaction sequence comprisign pressure-induced dissociation of the native enzyme into its subunits followed by subunit aggregation to form inactive high-molecular-weight particles.

Animals↗

PubMed1978

Oxidation of sulfhydryl groups in lactate dehydrogenase under high hydrostatic pressure.

Lactate dehydrogenase from rabbit skeletal muscle in the presence of substrate exhibits irreversible deactivation at hydrostatic pressures beyond 1 kbar [Schmid, G., Lüdemann, H.-D. & Jaenicke, R. (1975) Biophys. Chem. 3, 90-98]. In the absence of substrate and coenzyme the lability towards pressure is enhanced. The pH dependence of the effect and its inhibition by SH-protecting agents suggest the oxidation of sulfhydryl groups to be involved in the mechanism of deactivation. Partial deactivation observed at pH 5.5-7.0 becomes complete in the range of the intrinsic pK of cysteine; addition of dithiothreitol and/or EDTA protects the enzyme from complete deactivation, and leads to the residual enzymatic activity observed at pH 7.0. Incubation of the enzyme with dithiothreitol after pressure deactivation at pH 8.5 causes partial reactivation. From pressure-dependent measurements of the kinetics of deactivation an activation volume of deltaVnot equal to = -285 +/- 30 cm3 . mol-1 is calculated, which exceeds numerical data reported for typical reactions in organic chemistry. Therefore, the assumption can be made that the oxidation of sulfhydryl groups is connected with structural changes in the enzyme in the rate-determining step of the deactivation. The proposed mechanism may contribute to the toxicity of oxygen towards bacteria under high hydrostatic pressure.

Animals↗

PubMed1982

The influence of hydrostatic pressure and contralateral cooling on capillary blood cell velocity and transcutaneous oxygen tension in fingers.

Measurement of transcutaneous oxygen tension (pTcO2) has become a rather widely used technique. The relation of pTcO2 to skin microcirculatory parameters is, however, unclear. Capillary blood cell velocity (CBV) was compared with transcutaneous oxygen tension (pTcO2) of the finger in seven volunteers on change of hydrostatic pressure and contralateral cooling. On increase in hydrostatic pressure and also with contralateral cooling CBV decreased, while pTcO2 remained unchanged. This finding can be explained by a heat-induced local vasodilation under the pTcO2 probe in spite of a general increase in vasomotor tone. During decreasing perfusion pressure CBV was unchanged, while pTcO2 fell. This shows that with reduction in driving pressure skin pO2 is lowered. Teh absence of a decrease in CBV under these circumstances may be explained by a closure of the arteriovenous shunts, with diversion of blood into the superficial nutritional capillaries. It is suggested that CBV is related to changes in skin vasomotor tone. The pTcO2 findings indicate that the skin vasculature under the isothermally heated (44 degrees C) sensor is dilated and does not respond to vasoconstrictor stimuli.

Adult↗

PubMed1984

Effect of high hydrostatic pressure on sodium transport across toad skin.

The short-circuit current (Isc) was measured as an index of active net sodium transport across the isolated toad skin under various hydrostatic pressures up to 300 ATA. Upon compression, the base-line Isc increased transiently during the first 10 min by 10-15 microA/cm2 (congruent to 20%) and then decreased continuously, leveling off at 40 min under pressure. The latter decrease in base-line Isc (P less than 0.05 at all pressures) was pressure dependent, and its magnitude was 30 microA/cm2 (60% inhibition) at 300 ATA. Similarly, the transepithelial electric potential difference (PD) tended to increase slightly during the early phase of compression and decreased during the steady-state phase of compression. The transepithelial resistance (R), calculated from PD/Isc ratios, generally increased under pressure. The addition of vasotocin to the inside bathing medium resulted in an increase in Isc and PD and a reduction in R at all pressures. The magnitude of peak Isc response to vasotocin was 50-60 microA/cm2 at pressures up to 100 ATA, but decreased to 30 microA/cm2 at 200-300 ATA (0.05 less than P less than 0.10 as compared to the response at 1 ATA). On the other hand the stimulatory effect of 1 mM cyclic adenosine monophosphate (cAMP), added to the inside bathing medium, on Isc was not affected by pressure between 1 and 300 ATA. From these results it is postulated that the inhibition of base-line Na transport under high hydrostatic pressure may be primarily due to a decrease in the outer membrane permeability to Na rather than an inhibition of the Na-K-ATPase.

Animals↗

PubMed1998

Two-electrode voltage clamp of Xenopus oocytes under high hydrostatic pressure.

Functional assays of cloned ion channels and other transport systems under various hydrostatic pressures provide information on the apparent changes in protein volume occurring during conformational rearrangements. Thus, they are valuable tools in the detailed study of the molecular steps underlying the functioning of such proteins. Here we present details of a set-up which can be used for two-electrode voltage-clamp experiments on Xenopus oocytes, commonly used for heterologous protein expression, at hydrostatic (oil) pressures as high as 60 MPa (approximately 600 atm.). The advantages of this set-up over pneumatic systems include the minimization of compression/decompression-induced temperature changes, and an increased safety of handling due to the small volume (< 10 ml) of compression medium (oil) required. The performance of the system is illustrated using experimental data on the effects of high pressure on currents recorded from oocytes expressing a Shaker potassium channel mutant. This set-up is suitable for the investigation of all electrically measurable transport systems expressed in Xenopus oocytes.

Animals↗

PubMed1990

Effect of the tyrosine 96 hydrogen bond on the inactivation of cytochrome P-450cam induced by hydrostatic pressure.

The effects of removal of the tyrosine 96 hydrogen bond on the stability and conformational events of cytochrome P-450cam are presented in this communication. Hydrostatic pressure has been used as a tool to perturbe the structure leading to the formation of cytochrome P-420, an inactivated but soluble and undenatured form of the enzyme. We show that the spin transition of cytochrome P-450cam, which is known to be influenced by hydrostatic pressure, is affected by this single mutation. The free energy of stabilisation of native substrate-free cytochrome P-450cam is not affected by the removal of the tyrosine 96 hydrogen bond via mutagenesis to phenylalanine, whereas the substrate-bound protein shows a difference of 21 kJ/mol. These results, as well as an observed 110 ml/mol difference for the volume of the inactivation reaction between substrate-bound native and mutant proteins, have been interpreted in terms of a more hydrated heme pocket for the site-directed mutant at position 96 compared to the wild-type protein where camphor is tightly bound via the tyrosine 96 hydrogen bond and water excluded from the active site.

Camphor↗

PubMed2002

Conformational heterogeneity is revealed in the dissociation of the oligomeric chaperonin GroEL by high hydrostatic pressure.

We investigated the dissociation of tetradecameric GroEL by high hydrostatic pressure in the range of 1-2.5 kbar. Kinetics of the dissociation of GroEL in the absence and presence of Mg(2+) and/or KCl were monitored using light scattering. All of the kinetics were biphasic in nature. At any given pressure, only monomers and 14mers were produced, and below 2.5 kbar, the 14mers only partially dissociated to monomers, which did not significantly reassemble on depressurization. Under identical reaction conditions, the observed dissociation rates decreased by only 2-fold when the concentration of GroEL was increased by 20-fold. At 2.5 kbar the observed rates decreased exponentially with the increase in [KCl] and reached a minimum at approximately 75mM. Similarly, the rates decreased with the increase in [Mg(2+)] and reached a minimum at approximately 3 mM Mg(2+). In the presence of saturating amounts of Mg(2+) (10 mM) and KCl (100 mM), the rates were much faster than with 10 mM Mg(2+) alone. The results could be rationalized in terms of the presence of GroEL heterogeneity, which could not be assessed easily by common techniques such as sedimentation velocity, HPLC, gel electrophoresis, and dissociation by chaotropes. This heterogeneity is evidence of subpopulations of GroEL that dissociate at different pressures. At low pressures, the oligomer without added Mg(2+) only partially dissociates to monomers, leading to an apparent plateau in the kinetics, whereas in the presence of Mg(2+) the species are converted to a tighter Mg(2+)-bound species, leading to a much slower dissociation process. The presence of KCl in the sample also leads to similar heterogeneity.

Chaperonin 60↗

PubMed2005

Cellular growth under hydrostatic pressure using bovine aortic EC-SMC co-cultured ePTFE vascular graft.

High blood pressure (hypertension) is implicated in the development of atherosclerosis. Blood vessels are constantly subjected to stretch due to blood pressure and changes in stretch usually instigate adaptive vascular remodeling, including abnormal growth and proliferation of vascular smooth muscle cells (VSMCs) as well as extracellular matrix (ECM). In this experiment, we used bovine aortic endothelial cells and smooth muscle cells (EC-SMC) co-cultured ePTFE vascular grafts subjected to normal atmospheric pressure (as a control), and 100 mmHg hydrostatic pressure for 7 d. The increase of cell layer thickness was observed. When measured, the cell layer thickness increased by 116.2%. The increase of collagen (Type IV) synthesis was also observed in the immunohistochemistry assay. When stained with toluidine blue, the cells showed metachromatic phenomenon.

Animals↗

PubMedDate not supplied

[Effect of elevated hydrostatic pressure on the shape of human erythrocytes].

Influence of high hydrostatic pressure (2600-4300 atm) on the shape of human red blood cells was studied. An irreversible change of the shape was observed, which was similar to that seen in the case of a shift stress. The model modification dynamics of the erythrocyte shape was proposed. Activation energy and activation volume of the defects resulting in shape changes were calculated.

Erythrocytes↗

PubMed2006

Water activity of bacterial suspension media unable to account for the baroprotective effect of solute concentration on the inactivation of Listeria monocytogenes by high hydrostatic pressure.

Inactivation of Listeria monocytogenes (10(8) CFU/ml) by high hydrostatic pressure (HHP) from 400 to 600 MPa at 25 degrees C for 10 min was investigated with various concentrations of sodium chloride, sucrose, and sodium phosphate buffer solutions. Sodium chloride significantly inhibited HHP-induced inactivation of L. monocytogenes at concentrations higher than 2.6 M. A low concentration of sodium chloride within 1.7 M had no effect on HHP-induced inactivation. Almost complete inactivation at relatively low sodium chloride concentration solution was observed with treatments above 500 MPa. Sucrose also significantly inhibited HHP-induced inactivation of L. monocytogenes when greater than 1.2 M sucrose solutions were used. HHP-treatment at 400 MPa reduced the number of L. monocytogenes in 1.2 M, 1.5 M, and 1.8 M sucrose solutions by 4.8, 2.0, and 0.7 log cycles, respectively. Higher pressure did not yield significant reductions. Sodium phosphate buffer significantly inhibited HHP-induced inactivation of L. monocytogenes. In particular, 1 M phosphate buffer completely inhibited HHP-induced inactivation even at 600 MPa. HHP-treatment at 400 MPa reduced the number of L. monocytogenes in 0.1 M, 0.25 M, and 0.5 M phosphate buffer solutions by 5.6, 4.1, and 3.2 log cycles, respectively. The effect of HHP-induced inactivation of L. monocytogenes in the three kinds of solution was evaluated by adjusting water activity (a(w)). However, the baroprotective effect differed depending on the kind of solute even at the same a(w). This result showed no consistent correlation between a(w) and solute concentration in terms of the baroprotective effect. As an alternative approach, saturation of suspension solution was used for evaluating the effect of HHP-induced inactivation of L. monocytogenes. As the saturation of suspension media increased, the effect of HHP-induced inactivation of L. monocytogenes decreased regardless of the kinds of solute. The saturation of solution would be an alternative parameter of inhibition in terms of HHP-induced inactivation of bacteria.

Colony Count, Microbial↗

PubMed2003

SulA-independent filamentation of Escherichia coli during growth after release from high hydrostatic pressure treatment.

To improve the efficiency of sterilization by high hydrostatic pressure treatment (HPT), it is desirable to know the biochemical process of bacteria most sensitive to the treatment. We investigated growth properties after release from HPT of exponentially growing Escherichia coli K-12 cells. We observed growth retardation after treatment (30 min at 37 degrees C) above 75 MPa. Long filamentous cells of about eight times normal cell length were observed at 90 min growth after treatment at 75 MPa. In the subsequent period the filamentous cells divided into normal-sized cells. recA and sulA mutant strains also formed filamentous cells, indicating that filamentation was SulA-independent. Nucleoids segregated normally in the filamentous cells. Only one FtsZ ring (or none) was detected at possible division sites in the elongated cells. Western blotting analysis demonstrated that the amount of FtsZ protein was not affected by the treatment. GTP-dependent in vitro polymerization of either FtsZ protein in E. coli crude extract or purified FtsZ protein, however, was sensitive to HPT. These facts suggest that HPT at 75 MPa denatures a fraction of FtsZ molecules, and that these denatured molecules interfere with the polymerization of functional FtsZ, resulting in the significantly reduced number of FtsZ rings.

Cell Division↗

PubMed2000

Evaluation of batch and semicontinuous application of high hydrostatic pressure on foodborne pathogens in salsa.

The effects of high hydrostatic pressure (HPP; 545 MPa) on strains of Escherichia coli O157:H7, Listeria monocytogenes, enterotoxigenic Staphylococcus aureus, and nonpathogenic microorganisms were studied in tomato-based salsa. Products were evaluated for the survival of the inoculated pathogens following HPP treatment and after storage at 4 degrees C and 21 to 23 degrees C for up to 2 months. Inoculated samples without HPP treatment, stored under the same conditions, were also evaluated to determine the effects of the acid environment of salsa on the survival of inoculated strains. None of the inoculated pathogens were detected in the HPP-treated samples for all treatments throughout the storage period. Inoculated pathogens were detected in the non-HPP-treated samples stored at 4 degrees C after 1 month, with L. monocytogenes showing the highest level of survivors. In the non-HPP-treated samples stored at 21 to 23 degrees C, E. coli and S. aureus were not detected after 1 week, but L. monocytogenes was detected in low levels. Studies with nonpathogenic strains of the pathogens were conducted at Oregon State University using HPP treatments in a semicontinuous production system. The nonpathogenic microorganisms (E. coli, Listeria innocua, Listeria welshimeri, and nonenterotoxigenic S. aureus) were inoculated together into a feeder tank containing 100 liters of salsa. Microbiological results of samples collected before HPP treatment and from the aseptic filler were similar to those obtained for the pathogenic strains. No survivors were detected in any of the HPP-treated samples.

Bacteria↗

PubMed2005

A three-dimensional collagen matrix as a suitable culture system for the comparison of cyclic strain and hydrostatic pressure effects on intervertebral disc cells.

OBJECT: To study intervertebral disc cell mechanobiology, the authors developed experimental systems that allow the application of cyclic strain and intermittent hydrostatic pressure (IHP) on isolated disc cells under equal three-dimensional (3D) culture conditions. The purpose of the study was to characterize disc cell proliferation, viability, morphology, and gene expression in 3D collagen matrices. METHODS: The effects of cyclic strain (1, 2, 4, and 8% strain; 1 Hz) and IHP (0.25 MPa, 0.1 Hz) on gene expression (real-time polymerase chain reaction) of anabolic and catabolic matrix proteins were investigated and compared with those derived from mechanically unstimulated controls. Intervertebral disc cells proliferated in the collagen gels (mean viability 91.6%) and expressed messenger RNA for collagen I, collagen II, aggrecan, matrix metalloproteinase (MMP)-2, and MMP-3. Morphologically, both spindle-shaped cells with longer processes and rounded cells were detected in the collagen scaffolds. Cyclic strain increased collagen II and aggrecan expression and decreased MMP-3 expression of anulus fibrosus cells. No significant difference between the four strain magnitudes was found. Intermittent hydrostatic pressure tended to increase collagen I and aggrecan expression of nucleus cells and significantly decreased MMP-2 and -3 expression of nucleus cells, whereas aggrecan expression of anulus cells tended to decrease. CONCLUSIONS: Based on these results, the collagen matrix appeared to be a suitable substrate to apply both cyclic strain and IHP to intervertebral disc cells under 3D culture conditions. Individual variations may be influenced by the extent of degeneration of the disc specimens from which the cells were isolated. This experimental setup may be suitable for studying the influence of degeneration on the disc cell response to mechanical stimuli.

Aggrecans↗