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Effect of hydrostatic pressure on ligand binding to hemoglobin.

Increase in hydrostatic pressure to 1000 atm increased the affinity of human and menhaden (Brevoortia tyrannus) hemoglobins for oxygen. With necessary assumptions about the form of the equilibrium curve, and after correction for changes in pH and volume due to pressure, the increase in affinity is about 2-fold for both hemoglobins. At pH 6.5, Hill's n for menhaden hemoglobin is near 1, and it is believed to remain in the T state, whereas human hemoglobin undergoes a T to R transition. This suggests that the R-T equilibrium is not disturbed by pressure. In direct experiments the binding of a fluorescent effector (8 hydroxy-1,3,6-pyrene (trisulfonic acid) to deoxyhemoglobin was not changed by pressure. The binding of n-butylisocyanide to hemoglobin and to myoglobin is also greater at high pressures, similarly suggesting that the R-T transition is not involved in the pressure effect.

Animals↗

Effects of hydrostatic pressure on permeability of airway epithelium.

The presence of blood proteins and excess liquid in the airway lumen during airway inflammation may be secondary to extravasation and elevation of subepithelial hydrostatic pressure. This study examines how hydrostatic pressures of 5-20 cm H2O affect hydraulic conductivity and macromolecular permeability of primary cultures of bovine tracheal epithelium. Hydraulic conductivity was not altered by transepithelial pressure gradients of up to 20 cm H2O directed from the mucosal to serosal side of the tissue (m-s). By contrast, a 20-cm H2O s-m pressure resulted in a marked increase in hydraulic conductivity with the critical pressure lying between 10 and 20 cm H2O. Electrical conductance (i.e., permeability to ions) was not altered by m-s pressure gradients, or by a 5-cm H2O s-m gradient, but was increased by s-m pressures > or = 10 cm. Fluxes (s-m and m-s) of fluorescein and fluorescent dextrans (70 and 2000 kDa) were not altered by pressures of up to 20 cm H2O m-s. By contrast s-m pressure gradients of 20 cm H2O dramatically increased the s-m fluxes of these probes. The increases in flux were completely reversible. The results indicate that s-m pressure gradients greatly increase the hydraulic conductivity of airway epithelium by creating pores with an effective diameter greater than 54 nm.

Animals↗

Inactivation of Escherichia coli in milk by high-hydrostatic-pressure treatment in combination with antimicrobial peptides.

We studied the inactivation in milk of four Escherichia coli strains (MG1655 and three pressure-resistant mutants isolated from MG1655) by high hydrostatic pressure, alone or in combination with the natural antimicrobial peptides lysozyme and nisin and at different temperatures (10 to 50 degrees C). Compared with that of phosphate buffer, the complex physicochemical environment of milk exerted a strong protective effect on E. coli MG1655 against high-hydrostatic-pressure inactivation, reducing inactivation from 7 logs at 400 MPa to only 3 logs at 700 MPa in 15 min at 20 degrees C. An increase in lethality was achieved by addition of high concentrations of lysozyme (400 microg/ml) and nisin (400 IU/ml) to the milk before pressure treatment. The additional reduction amounted maximally to 3 logs in skim milk at 550 MPa but was strain dependent and significantly reduced in 1.55% fat and whole milk. An increase of the process temperature to 50 degrees C also enhanced inactivation, particularly for the parental strain, but even in the presence of lysozyme and nisin, a 15-min treatment at 550 MPa and 50 degrees C in skim milk allowed decimal reductions of only 4.5 to 6.9 for the pressure-resistant mutants. A substantial improvement of inactivation efficiency at ambient temperature was achieved by application of consecutive, short pressure treatments interrupted by brief decompressions. Interestingly, this pulsed-pressure treatment enhanced the sensitivity of the cells not only to high pressure but also to the action of lysozyme and nisin.

Animals↗

Estimate of the subepithelial hydrostatic pressure that drives inflammatory transudate into the airway lumen.

Inflammatory diseases of the upper respiratory tract are characterized by flow of plasma filtrate across the epithelium into the airway lumen ("transudation"). Elsewhere, we have proposed that extravasation from microvessels causes edema, and this is associated with elevated subepithelial hydrostatic pressure that drives transudation. To test this hypothesis, we have attempted to block transudation by elevating luminal hydrostatic pressure. We measured the appearance of plasma markers into the lumen of an isolated perfused segment of rat trachea in vivo and found that stimulation of one vagal nerve caused a rapid (half-time approximately 5 min) and nonselective increase in the flow of markers from blood to airway lumen. Leukocyte migration also caused transudation that developed much more slowly (half-time = 2-3 h). In both cases, transudation was blocked by application of luminal hydrostatic pressures. The critical luminal pressure needed to block vagally induced transudation was approximately 4.5 cmH2O, and, to block epithelial transudation induced by leukocyte traffic, it was 3 cmH2O, and we conclude that these are the subepithelial pressures that drive inflammatory transudation into the airway lumen.

Albumins↗

Effects of hydrostatic pressure on the Leptospira interrogans: high immunogenicity of the pressure-inactivated serovar hardjo.

The Hardjoprajitno strain of Leptospira interrogans serovar hardjo was subjected to different hydrostatic pressures. Complete inactivation occurred when the leptospires were treated with 2 kbar for 60 min. Electron microscopy showed dislocation of the outer membrane, partial loss of the helical shape and extrusion of the axial filament from the cytoplasmic cylinder of the pressurized leptospires. When the pressure-treated leptospires were inoculated into rabbits they were highly immunogenic. The sera of these animals presented a titer of 2048 in the microscopic serum agglutination reaction. Fluorescence measurements indicated that the action of pressure on the leptospires might have resulted from perturbation on membrane protein components, permitting the binding of the fluorescent probe bis (8-anilinonaphthalene-1-sulfonate) (Bis-ANS). This is the first report of the use of hydrostatic pressure to inactivate pathogenic bacteria with the potential to lead to a vaccine.

Anilino Naphthalenesulfonates↗

Interactions between permeability and hydrostatic pressure in perfused dogs' lungs.

To evaluate interactions between increased permeability of the pulmonary vasculature and increased hydrostatic pressure on lung fluid flux, we measured water and blood contents of five equally thick slices of in situ perfused dogs' lungs that had been vertically suspended and, after experimental manipulations, rapidly frozen. Measurements were made under four sets of conditions (4 dogs each): 1) control; 2) increased left atrial pressure (Pla), 25 cmH2O relative to the bottom of the lung; 3) oleic acid; and 4) oleic acid and increased Pla. Significant interactions on lung water content were found between 1) oleic acid and increased Pla (P less than 0.005) and 2) oleic acid and height of the slice of lung (P less than 0.003). Blood content was significantly affected by interactions between 1) oleic acid and height of the lung slice (P less than 0.001) and 2) increased Pla and height of the lung slice (P less than 0.001). We conclude that there are significant interactions between increased permeability and hydrostatic pressure that affect regional lung water content and intrapulmonary distribution of blood.

Animals↗

Gene expression profiles in chondrosarcoma cells subjected to cyclic stretching and hydrostatic pressure. A cDNA array study.

Mechanical forces have a profound effect on cartilage tissue and chondrocyte metabolism. Strenuous loading inhibits the cellular metabolism, while optimal level of loading at correct frequency raises an anabolic response in chondrocytes. In this study, we used Atlas Human Cancer cDNA array to investigate mRNA expression profiles in human chondrosarcoma cells stretched 8% for 6 hours at a frequency of 0.5 Hz. In addition, cultures were exposed to continuous and cyclic (0.5 Hz) 5 MPa hydrostatic pressure. Cyclic stretch had a more profound effect on the gene expression profiles than 5 MPa hydrostatic pressure. Several genes involved with the regulation of cell cycle were increased in stretched cells, as well as mRNAs for PDGF-B, glucose-1-phosphate uridylyltransferase, Tiam1, cdc37 homolog, Gem, integrin alpha6, and matrix metalloproteinase-3. Among down-regulated genes were plakoglobin, TGF-alpha, retinoic acid receptor-alpha and Wnt8b. A smaller number of changes was detected after pressure treatments. Plakoglobin was increased under cyclic and continuous 5 MPa hydrostatic pressure, while mitogen-activated protein kinase-9, proliferating cell nuclear antigen, Rad6, CD9 antigen, integrins alphaE and beta8, and vimentin were decreased. Cyclic and continuous pressurization induces a number of specific changes. In conclusion, a different set of genes were affected by three different types of mechanical stimuli applied on chondrosarcoma cells.

Bone Neoplasms↗

Compressibility of the heme pocket of substrate analogue complexes of cytochrome P-450cam-CO. The effect of hydrostatic pressure on the Soret band.

The effect of hydrostatic pressure on the electronic absorption spectrum of the carbon monoxide complex of cytochrome P-450cam (CYP101) in the presence of various substrates was studied. With increasing pressure the wavenumber of the Soret band in the cytochrome P-450-CO complex shifts linearily to lower values (red-shift) and the half-width increases (broadening). The microscopic theory of solvent-solute interaction discussed by Laird and Skinner is used to explain the observed pressure effects. According to this theory, the slope of the red-shift of the Soret band is related to the compressibility of the chromophore environment, that is the heme moiety of the hemoproteins. It was found that the slope of the red-shift and the slope of the broadening of the Soret band for the CO complex in the presence of various substrate analogues increase with the decrease of the initial high-spin content at 0.1 MPa in the oxidized state. Variation of the high-spin content reflects the changes in the number of water molecules and/or changes in the polarity of the heme environment. The higher compressibility of the cytochrome P-450 complexes with the substrate analogues, which induce a lower degree of the high-spin content in the oxidized protein, is explained by the ability of the water molecules in the heme moiety to transmit the pressure effect on the protein structure to the heme chromophore. Therefore, a larger pressure-induced red-shift of the Soret band in the CO complex of cytochrome P-450cam might indicate a higher water content in the heme environment.

Camphor 5-Monooxygenase↗

Re-evaluation of hydrostatic pressure treatment for malignant bladder lesions.

We reviewed 50 cases of bladder cancer treated with the hydrostatic pressure technique. Results were encouraging when 1) single or multiple papillary tumors were not located around the ureteral orifice, with a presumed depth of tumor infiltration within T2, I) there was no history of open operations, 3) there was observed activity of immunological surveillance, for example a positive reaction to a tuberculin test, and 4) management of anesthesia was satisfactory. Hydrostatic pressure therapy is not palliative treatment for far-advanced cases of bladder cancers but should be the first choice for new cases beyond the scope for transurethral operations and when indications in tumor and host condition are satisfied.

Blood Circulation↗

Variation in resistance to hydrostatic pressure among strains of food-borne pathogens.

Among food-borne pathogens, some strains could be resistant to hydrostatic pressure treatment. This information is necessary to establish processing parameters to ensure safety of pressure-pasteurized foods (N. Kalchayanand, A. Sikes, C. P. Dunne, and B. Ray, J. Food Prot. 61:425-431, 1998). We studied variation in pressure resistance among strains of Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella species at two temperatures of pressurization. Early-stationary-phase cells in 1% peptone solution were pressurized at 345 MPa either for 5 min at 25 degrees C or for 5, 10, or 15 min at 50 degrees C. The viability loss (in log cycles) following pressurization at 25 degrees C ranged from 0.9 to 3.5 among nine L. monocytogenes strains, 0.7 to 7.8 among seven S. aureus strains, 2.8 to 5.6 among six E. coli O157:H7 strains, and 5.5 to 8.3 among six Salmonella strains. The results show that at 25 degrees C some strains of each species are more resistant to pressure than the others. However, when one resistant and one sensitive strain from each species were pressurized at 345 MPa and 50 degrees C, the population of all except the resistant S. aureus strain was reduced by more than 8 log cycles within 5 min. Viability loss of the resistant S. aureus strain was 6.3 log cycles even after 15 min of pressurization. This shows that strains of food-borne pathogens differ in resistance to hydrostatic pressure (345 MPa) at 25 degrees C, but this difference is greatly reduced at 50 degrees C. Pressurization at 50 degrees C, in place of 25 degrees C, will ensure greater safety of foods.

Bacteria↗

Stability of bovine cytochrome C oxidase as studied after exposure to high hydrostatic pressure.

Structural and functional stability of bovine cytochrome c oxidase as a function of exposure to high hydrostatic pressure is reported. The pressure affects the stability of monomeric and dimeric enzyme quite differently. Exposure of the monomeric cytochrome c oxidase to pressures higher than 2.5 kbar causes dissociation of subunits III, VIa, VIb, VIIa with a 35-50% decrease in electron transport activity. Dimeric enzyme is more resistant to high hydrostatic pressure since subunits III and VIIa do not dissociate and the electron transport activity loss is minimal.

Animals↗

Combined influence of temperature and hydrostatic pressure on cardiac conduction.

Interactions of temperature and hydrostatic pressure were examined in three types of cardiac tissue. Decreases in temperature from 37 degrees C to 27 degrees C and increases in pressure from 1 to 150 ATA slowed conduction significantly in rabbit atria, dog atria, and dog Purkinje fibers. The combination of lowest temperature and highest pressure produced the greatest decreases in conduction in all three tissues. Purkinje fiber action potentials revealed that slowing of conduction can be attributed to depressed excitability, decreased membrane potential, and reduced maximum upstroke velocity of the action potential. These action potential variables exhibited the greatest change at the lowest temperature and highest pressure. Cooling or elevations in pressure increased the duration of the action potential. At 27 degrees C, however, the duration was unaffected by increases in pressure. Aberrant conduction developed occasionally, at 27 degrees C and 150 ATA, and could be explained by changes in action potential variables. The significance of this finding is discussed with respect to the possible development of cardiac arrhythmias in divers exposed to cold hyperbaric stress.

Action Potentials↗

Effect of hydrostatic pressure on spectra of heme compounds.

Increase in hydrostatic pressure shifts the absorption bands of oxy-, carboxy-, and deoxyhemoglobin and myoglobin toward the red by 0.4 to 0.7 nm corresponding to a change in extinction coefficient of from 4 to 8% at the peak of the difference spectrum. The pressure difference spectrum for oxyhemoglobin closely resembles the difference spectrum described by Adams and Schuster ((1974) Biochem. Biophys. Res. Commun. 58, 528-533) following addition of inositol hexaphosphate to oxyhemoglobin. A similar shift was observed for derivatives of dimethyl-deuterohemedisulfonate in both Fe2+ and Fe3+ forms indicating that the protein is not required for this effect, in contrast to earlier reports of T. L. Fabry and J. W. Hunt ((1968) Arch. Biochem. Biophys. R3, 428-429) and Q.H. Gibson and F.G. Carey ((1975) Biochem. Biophys. Res. Commun. 67, 747-571) who were unable to observe changes in aqueous solutions of protoheme derivatives.

Animals↗

Biphasic inactivation kinetics of Escherichia coli in liquid whole egg by high hydrostatic pressure treatments.

Kinetic studies on the isothermal high hydrostatic pressure (HHP) inactivation of Escherichia coli in liquid whole egg (LWE) were performed at 5 and 25 degrees C in the pressure range of 250-400 MPa. The characteristic tailing inactivation curves were described by a first-order biphasic model. As compared to a previous rheological study, it is suggested that the phase change of LWE during pressure treatment affects the inactivation rate of E. coli. Within the processing criteria where the rheological properties of LWE were still comparable to those of fresh LWE, HHP treatments at 5 degrees C induced more E. coli inactivations than those at 25 degrees C. From the results of approximately 3 log reductions of E. coli and over 5 log reductions of Pseudomonas and Paenibacillus, HHP treatment of LWE at 5 degrees C is regarded to be as effective as conventional thermal pasteurization. However, no post-process contamination and the consistency of temperature during preparation, HHP treatment, and storage provide clear processing advantages.

Colony Count, Microbial↗

[Effects of hydrostatic pressure on cardiac function in human].

Effects of hydrostatic pressure on cardiac function were measured by echocardiography and radiography. Seven normal subjects (three females and four males) aged from 21 to 45 years were performed echocardiography on long axis of the heart at standing position in air and in head-out water (36 degrees C) immersion. A same trials were done at long sitting position in four subjects (one female and three males). The measurement of cardiac functions was calculated by Pombo's method. A chest radiography of a normal male individual (45 years) was taken at the time of functional residual capacity of lung in air and in head-out water (36 degrees C) immersion. For taking the chest radiography in water and in air, a box made by wood was used. X-ray exposure was synchronized with R wave of electro-cardiogram. The end-diastolic left ventricular volume, end-systolic volume and cardiac output in water immersion were significantly increased to compare with that of in air. Mean +/- SD of these percent changes were 213 +/- 71% (p less than 0.005), 193 +/- 90% (p less than 0.05), 220 +/- 83% (p less than 0.005), respectively. A same results were obtained at long sitting position. There was no significant changes on the heart rate between in water and in air. The circumstance of the abdomen decreased significantly in water immersion in comparison to in air. The percent change was 94.4 +/- 3.1% (p less than 0.001). There was a significant correlation between percent changes of abdominal circumstance and percent changes of end-systolic left ventricular volume (r = 0.754, p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Selective expression of neural cell adhesion molecule (NCAM)-180 in optic nerve head astrocytes exposed to elevated hydrostatic pressure in vitro.

Glaucomatous optic neuropathy is usually associated with elevated intraocular pressure. Optic nerve head astrocytes may respond to intraocular pressure by stimulation of pressure-sensitive mechanoreceptors on the cell surface. Neural cell adhesion molecule (NCAM) a transmembrane protein, mediates cell adhesion and migration. The NCAM 180 isoform increases in astrocytes of glaucomatous optic nerve head. We characterized the relative expression of NCAM isoforms in human optic nerve head astrocytes grown under elevated hydrostatic pressure. Astrocytes cultured from normal human optic nerve heads were exposed to either atmospheric or continuous hydrostatic pressure of 60 mm Hg, and analyzed at 6-48 h. Changes in cell shape, immunoreactivity, and distribution of GFAP, actin and NCAM were observed in pressure-treated cultures. Newly synthesized (35)S-labeled NCAM protein immunoprecipitated from cell lysates was increased 2-fold within 24 h after exposure to elevated pressure compared to control. The increase in NCAM synthesis was primarily due to the NCAM 180 isoform. A significant increase in NCAM 180 mRNA levels was detected by RT-PCR and Northern blots in cultured optic nerve head astrocytes within 6 h after exposure to elevated pressure. NCAM 180 mRNA and protein synthesis decreased after 24 h and returned to control levels by 48 h. Our data indicate that NCAM 180 transcription and synthesis in astrocytes is stimulated by elevated hydrostatic pressure. Because NCAM 180 interacts with the cytoskeleton through an extended cytoplasmic tail, a selective and transient increase in NCAM 180 in optic nerve head astrocytes exposed to elevated pressure may be relevant to the migration and interactions of reactive astrocytes in glaucoma.

Adolescent↗

High hydrostatic pressure as a tool to study protein aggregation and amyloidosis.

Aggregation of proteins is a serious problem, affecting both industrial production of proteins and human health. Despite recent advances in the theories and experimental techniques available to address understanding of protein aggregation processes, mechanisms of aggregate formation have proved challenging to study. This is in part because the typical irreversibility of protein aggregation processes at atmospheric conditions complicates analysis of their kinetics and thermodynamics. Because high hydrostatic pressures act to disfavor the hydrophobic and electrostatic interactions that cause protein aggregation, studies conducted under high hydrostatic pressures may allow protein aggregates to be formed reversibly, enabling thermodynamic and kinetic parameters to be measured in greater detail. Although application of high hydrostatic pressures to protein aggregation problems is rather recent, a growing literature, reviewed herein, suggests that high pressure may be a useful tool for both understanding protein aggregation and reversing it in industrial applications.

Amyloidosis↗