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PubMed1988

Effect of increased hydrostatic pressure on lymphatic elimination of hyaluronan from sheep lung.

The effects of increased hydrostatic pressure on the concentrations of hyaluronan (hyaluronic acid) in lung lymph and serum were investigated in awake sheep with a cannula in the efferent vessel from the caudal mediastinal lymph node. Lung lymph was sampled at base line [left atrial pressure (LAP) 6.5 +/- 1.7 mmHg] and after two increases of LAP to 25.7 +/- 2.2 mmHg (level 1) and 37.0 +/- 5.1 mmHg (level 2). The lung lymph flow increased from 1.9 +/- 0.5 at base line to 9.3 +/- 2.2 and 15.9 +/- 0.7 ml/30 min, and the lymph-to-plasma concentration ratio of total protein decreased from 0.63 +/- 0.02 to 0.32 +/- 0.04 and 0.32 +/- 0.05 at the two elevated levels of LAP, respectively. The hyaluronan concentration in lung lymph was unchanged, and there was a flow-dependent elimination of hyaluronan from the lung that increased from 23 +/- 8 to 87 +/- 19 and 137 +/- 37 micrograms/30 min, respectively. The lung concentration of hyaluronan was 167 +/- 28 micrograms/g fresh lung, and at base line it was calculated that slightly less than 2% of the lung hyaluronan was eliminated by the lymphatic route in 24 h. If extrapolated to 24 h, the elimination rate of hyaluronan seen during elevated LAP would result in lymphatic elimination of 18% of the lung hyaluronan over this time period. Since hyaluronan is responsible for part of the protein exclusion in the extracellular matrix, it is plausible that washout of interstitial hyaluronan contributes to the decrease in albumin exclusion from the interstitium that occurs after an elevation of LAP.

Animals↗
PubMed1978

Hydrostatic pressure effects upon cellular leakage and active transport by Vibrio marinus.

Hydrostatic pressures greater than ca. 400 atm cause release of cellular components of Vibrio marinus MP-1 in the order protein greater than RNA greater than malate dehydrogenase greater than DNA greater than amino acids. Increased pressures also slow the rate of cycloleucine (non-metabolizable analogue of L-leucine acid L-valine) uptake but increase its efflux rate. Lineweaver-Burk plots (1/V versus 1/[S]) of cycloleucine uptake indicate that the Km increases with compression which suggests a decrease in affinity of the transport system for substrate at increased pressures.

Amino Acids↗
PubMed1987

Therapeutic vaccination against adjuvant arthritis using autoimmune T cells treated with hydrostatic pressure.

An ideal treatment for autoimmune diseases would be a nontoxic means of specifically neutralizing the autoreactive lymphocytes responsible for the disease. This goal has been realized in experimental autoimmunity models by immunizing rats or mice against their own autoimmune cells such that the animals generate an immune response specifically repressive to the disease-producing lymphocytes. This maneuver, termed lymphocyte vaccination, was demonstrated to be effective using some, but not all, autoimmune helper T-lymphocyte lines. We now report that T lymphocytes, otherwise incapable of triggering an immune response, can be transformed into effective immunogens by treating the cells in vitro with hydrostatic pressure. Clone A2b, as effector clone that recognized cartilage proteoglycan and caused adjuvant arthritis in Lewis rats, is such a cell. Untreated A2b could not trigger an immune response, but inoculating rats with pressure-treated A2b induced early remission of established adjuvant arthritis as well as resistance to subsequent disease. Specific resistance to arthritis was associated with anti-idiotypic T-cell reactivity to clone A2b and could be transferred from vaccinated rats to naive recipients using donor lymphoid cells. Aggregation of T-lymphocyte membrane components appeared to be important for an immune response because the effects of hydrostatic pressure could be reproduced by treatment of A2b with chemical cross-linkers or with agents disrupting the cytoskeleton. Populations of lymph node cells from antigen-primed rats, when treated with hydrostatic pressure, could also induce suppression of disease. Thus, effective vaccines can be developed without having to isolate the autoimmune T lymphocytes as lines or clones. These results demonstrate that effector T lymphocytes suitably treated may serve as agents for specifically controlling the immune system.

Animals↗
PubMed2004

Inactivation of Colletotrichum gloeosporioides spores by high hydrostatic pressure combined with citral or lemongrass essential oil.

Anthracnose, caused by the fungus Colletotrichum gloeosporioides, is the main post-harvest disease of the papaya. Inactivation of the spores of C. gloeosporioides in saline solution by the use of high hydrostatic pressure, citral oil and lemongrass oil, alone and in combination, was studied. C. gloeosporioides spores were efficiently inhibited after a pressure treatment of 350 MPa for 30 min. When C. gloeosporioides was treated with 0.75 mg ml(-1) of citral or lemongrass oil, the pressure needed to achieve the same spore inhibition was 150 MPa. This work suggests the use of high hydrostatic pressure and plant essential oils as an alternative control for fruit diseases.

Acyclic Monoterpenes↗
PubMed1991

The effect of hydrostatic pressure on the interaction of actomyosin subfragment 1 with nucleotides.

Increased hydrostatic pressure has previously been shown to reduce the tension of isometrically contracting skinned muscle fibres. An isomerization of the actomyosin complex is known to be pressure sensitive, but the pressure sensitivity of other steps in the ATPase pathway has not been characterised. We report here the effect of pressure on the ATP hydrolysis step of the myosin subfragment 1 ATPase, ADP binding to actomyosin subfragment 1 and the rate of ATP induced dissociation of actomyosin subfragment 1. We discuss the relationship of these changes to the observed effect of pressure on skinned muscle fibres.

Actomyosin↗
PubMed1999

Hydrostatic pressure rescues native protein from aggregates.

Misfolding and misassembly of proteins are major problems in the biotechnology industry, in biochemical research, and in human disease. Here we describe a novel approach for reversing aggregation and increasing refolding by application of hydrostatic pressure. Using P22 tailspike protein as a model system, intermediates along the aggregation pathway were identified and quantitated by size-exclusion high-performance liquid chromatography (HPLC). Tailspike aggregates were subjected to hydrostatic pressures of 2.4 kbar (35,000 psi). This treatment dissociated the tailspike aggregates and resulted in increased formation of native trimers once pressure was released. Tailspike trimers refolded at these pressures were fully active for formation of infectious viral particles. This technique can facilitate conversion of aggregates to native proteins without addition of chaotropic agents, changes in buffer, or large-scale dilution of reagents required for traditional refolding methods. Our results also indicate that one or more intermediates at the junction between the folding and aggregation pathways is pressure sensitive. This finding supports the hypothesis that specific determinants of recognition exist for protein aggregation, and that these determinants are similar to those involved in folding to the native state. An increased understanding of this specificity should lead to improved refolding methods.

Chromatography, Gel↗
PubMed2005

Induction of oxidative stress by high hydrostatic pressure in Escherichia coli.

Using leaderless alkaline phosphatase as a probe, it was demonstrated that pressure treatment induces endogenous intracellular oxidative stress in Escherichia coli MG1655. In stationary-phase cells, this oxidative stress increased with the applied pressure at least up to 400 MPa, which is well beyond the pressure at which the cells started to become inactivated (200 MPa). In exponential-phase cells, in contrast, oxidative stress increased with pressure treatment up to 150 MPa and then decreased again, together with the cell counts. Anaerobic incubation after pressure treatment significantly supported the recovery of MG1655, while mutants with increased intrinsic sensitivity toward oxidative stress (katE, katF, oxyR, sodAB, and soxS) were found to be more pressure sensitive than wild-type MG1655. Furthermore, mild pressure treatment strongly sensitized E. coli toward t-butylhydroperoxide and the superoxide generator plumbagin. Finally, previously described pressure-resistant mutants of E. coli MG1655 displayed enhanced resistance toward plumbagin. In one of these mutants, the induction of endogenous oxidative stress upon high hydrostatic pressure treatment was also investigated and found to be much lower than in MG1655. These results suggest that, at least under some conditions, the inactivation of E. coli by high hydrostatic pressure treatment is the consequence of a suicide mechanism involving the induction of an endogenous oxidative burst.

Cytoplasm↗
PubMed2002

Marinitoga piezophila sp. nov., a rod-shaped, thermo-piezophilic bacterium isolated under high hydrostatic pressure from a deep-sea hydrothermal vent.

A thermophilic, anaerobic, piezophilic, chemo-organotrophic sulfur-reducing bacterium, designated as KA3T, was isolated from a deep-sea hydrothermal chimney sample collected at a depth of 2630 m on the East-Pacific Rise (13 degrees N). When grown under elevated hydrostatic pressure, the cells are rod-shaped with a sheath-like outer structure, motile, have a mean length of 1-1.5 microm and stain Gram-negative. They appear singly or in short chains. When grown at lower, or atmospheric, pressures, the cells elongate and become twisted. Growth is enhanced by hydrostatic pressure; the optimal pressure for growth is 40 MPa (26 MPa pressure at sampling site). The temperature range for growth is 45-70 degrees C, the optimum being around 65 degrees C (doubling time is approximately 20 min at 40 MPa). Growth is observed from pH 5 to pH 8, the optimum being at pH 6. The salinity range for growth is 10-50 g NaCl l(-1), the optimum being at 30 g l(-1). The isolate is able to grow on a broad spectrum of carbohydrates or complex proteinaceous substrates, and growth is stimulated by L-cystine and elemental sulfur. The G+C content of the genomic DNA is 29 +/- 1 mol%. According to phylogenetic analysis of the 16S rDNA gene, the strain is placed within the order Thermotogales, in the bacterial domain. On the basis of 16S rDNA sequence comparisons and morphological, physiological and genotypic characteristics, it is proposed that the isolate be described as a novel species of the genus Marinitoga, with Marinitoga piezophila sp. nov. as the type species. The type strain is KA3T (= DSM 14283T = JCM 11233T).

Culture Media↗
PubMed2005

Temperature and treatment time influence high hydrostatic pressure inactivation of feline calicivirus, a norovirus surrogate.

Interest in high hydrostatic pressure processing as a nonthermal pasteurization process for foods continues to increase. Feline calicivirus (FCV), a propagable virus that is genetically related to the nonpropagable human noroviruses, was used for detailed evaluation of the high pressure processing parameters necessary for virus inactivation. Pressure inactivation curves of FCV strain KCD in Dulbecco's modified Eagle medium with 10% fetal bovine serum were obtained at 200 and 250 MPa as a function of time at room temperature. Pressure inactivation curves at 200 and 250 MPa also were determined as a function of temperature ranging from --10 to 50 degrees C at treatment times of 4 and 2 min, respectively. Tailing was observed for inactivation as a function of treatment time, indicating that the linear model was not adequate for describing these curves. The two nonlinear models, the log logistic and Weibull functions, consistently produced better fit to inactivation curves than did the linear model. The mean square errors were 0.381 for the log logistic model, 0.425 for the Weibull model, and 1.546 for the linear model. For inactivation as a function of temperature, FCV was most resistant to pressure at 20 degrees C. Temperatures above and below 20 degrees C significantly increased pressure inactivation of FCV. A 4-min treatment of 200 MPa at --10 and 50 degrees C reduced the titer of FCV by 5.0 and 4.0 log units, respectively; whereas at 20 degrees C the same treatment only reduced the titer by 0.3 log units. These novel results point to the potential for using temperatures above and particularly below room temperature to lower the pressure needed to cause the desired level of virus inactivation.

Animals↗
PubMedDate not supplied

Hydrostatic pressure measurement of endolymph and perilymph in the guinea pig cochlea.

Hydrostatic pressures of endolymph and perilymph were measured with a servo-micropipet system. The validity of the experiment was assessed by observation of endolymphatic pressure changes during and after 3 minutes of anoxia. Simultaneous recording of endocochlear DC potential confirmed the location of the electrode tip. Following verification of the efficacy of the system, measurements of endolymphatic or perilymphatic pressures were made in ten guinea pigs in which the middle ears had been filled with a saturated sodium chloride solution. Both endolymphatic and perilymphatic measurements showed gradual declines in pressure, with minimum pressure at 15 to 20 minutes and slow recovery during the next 20 minutes.

Animals↗
PubMed1985

Mechanisms of inhibition of (Na,K)-ATPase by hydrostatic pressure studied with fluorescent probes.

To investigate the mechanisms by which hydrostatic pressure inhibits (Na,K)-ATPase, we measured enzyme activity, as a function of pressure and temperature, of purified (Na,K)-ATPase from dog kidney and eel electroplax, and we monitored protein conformation, possible subunit interactions, and the fluidity of the membrane with fluorescent probes. The (Na,K)-ATPase and p-nitrophenylphosphatase activities were inhibited reversibly by pressures below 1.5 kilobars (eel enzyme) and 2.5 kilobars (dog kidney enzyme). Above these pressures, the enzymes were inactivated irreversibly. The plots of 1n(activity) versus pressure were curvilinear; this indicates that the reversible inhibition by pressure involves two or more rate-limiting steps. The calculated activation volumes varied with temperature and pressure and were larger for the (Na,K)-ATPase activity compared to the p-nitrophenylphosphatase activity. The fluorescence polarization of three hydrophobic probes decreased with increasing temperature (10-36 degrees C) and increased with increasing pressure (10(-3)-1.5 kilobars), reversibly, without any evidence of a lipid phase transition. Plots of enzyme activity versus fluorescence polarization of the lipid probes showed an inverse relationship; this indicates that enzyme activity was directly related to the fluidity of the membrane as measured by the lipid probes. Pressure had no effect on the fluorescence polarization of two cardiac glycoside probes nor on the efficiency of resonance energy transfer between either donor and acceptor cardiac glycosides specifically bound to the ouabain sites of different alpha-subunits, or tryptophan and the bound cardiac glycoside probe. These results suggest that dissociation of dimeric alpha-subunits is not related to the inhibition by pressure, and that the cardiac glycoside-complexed enzyme conformation is stabilized by pressure. It is concluded that increased pressure decreases the membrane fluidity which hinders conformational transitions associated with rate-limiting steps of the (Na,K)-ATPase reaction. It is proposed that ion-bound or -occluded forms of (Na,K)-ATPase are stabilized by pressure because they occupy a smaller volume.

Animals↗
PubMed2004

Analysis of hydrostatic pressure effects on transcription in Escherichia coli by DNA microarray procedure.

Hydrostatic pressure is a well-known physical stimulus, but its effects on cell physiology have not been clarified. To investigate pressure effects on Escherichia coli, we carried out DNA microarray analysis of the entire E. coli genome. The microarray results showed pleiotropic effects on gene expression. In particular, heat- and cold-stress responses were induced simultaneously by the elevated pressure. Upon temperature stress (including both temperature up- and down-shifts) and other environmental stresses, gene expression adjusts to adapt to such environmental changes through regulations by several DNA-binding proteins. An E. coli mutant, which deleted the hns gene encoding one of the regulator proteins, exhibited great pressure sensitivity. The result suggested that the H-NS protein was a possible transcriptional regulator for adaptation of the high-pressure stress.

Bacterial Proteins↗
PubMed1998

Joint fluid hydrostatic pressures that empty synovial capillaries of red cells have a wide range and correlate with pressures emptying arterioles.

OBJECTIVE: To study if, as predicted by calculations based on 3 Starling pressures, the fourth one, capillary pressure, may increase enough to contribute to joint swelling. METHODS: At magnifying arthroscopy, red cells mostly flow separated from each other in capillaries and precapillary vessels (arterioles). These are emptied of erythrocytes at intraarticular injection of saline to "tamponade" pressures. Patients sent for arthroscopy of the knee joint for mechanical or other lesions, or "internal derangement" (n = 30) or osteoarthritis (n = 26) were studied. RESULTS: The range of capillary (including postcapillary venular) tamponade pressure was 9-70 mm Hg. It correlated (r = 0.536, p < 0.001) with arteriolar tamponade pressure (range 20-100 mm Hg). CONCLUSION: If intravascular hydrostatic pressure is the main determinant of tamponade pressure, then increase of capillary pressure, possibly due to arteriolar dilatation induced precapillary pressure increase, may be a key mechanism in enhanced formation of synovial fluid.

Adolescent↗
PubMed1999

Thermococcus barophilus sp. nov., a new barophilic and hyperthermophilic archaeon isolated under high hydrostatic pressure from a deep-sea hydrothermal vent.

A novel barophilic, hyperthermophilic, anaerobic sulfur-metabolizing archaeon, strain MPT (T = type strain), was isolated from a hydrothermal vent site (Snakepit) on the Mid-Atlantic Ridge (depth, 3550 m). Enrichments and isolation were done under 40 MPa hydrostatic pressure at 95 degrees C. Strain MPT was barophilic at 75, 80, 85, 90, 95 and 98 degrees C, and was an obligate barophile between 95 and 100 degrees C (Tmax). For growth above 95 degrees C, a pressure of 15.0-17.5 MPa was required. The strain grew at 48-95 degrees C under atmospheric pressure. The optimal temperature for growth was 85 degrees C at both high (40 MPa) and low (0.3 MPa) pressures. The growth rate was twofold higher at 85 degrees C under in situ hydrostatic pressure compared to at low pressure. Strain MPT cells were motile, coccoid, 0.8-2.0 microns in diameter and covered by a hexagonal S-layer lattice. The optimum pH and NaCl concentration for growth at low pressure were 7.0 and 20-30 g l-1, respectively. The new isolate was an obligate heterotroph and utilized yeast extract, beef extract and peptone for growth. Growth was optimal in the presence of elemental sulfur. Rifampicin and chloramphenicol inhibited growth. The core lipids consisted of a major archaeol and a complex lipid pattern consisting of a major phospholipid. The DNA G + C content was 37.1 mol%. Sequencing of the 16S rRNA gene revealed that strain MPT belonged to the genus Thermococcus and it is proposed that this isolate should be designated as a new species, Thermococcus barophilus.

Atlantic Ocean↗
PubMed1976

Trying to explain an effect of "per se" hydrostatic pressure on heart rate in fish.

Specific effects of "per se" hydrostatic pressure on mean heart rate have been studied on eels (Anguilla anguilla L.), both untreated and treated with atropine and propranolol, and on isolated eel's heart. Because temperature brings, by itself, heart rate modifications in fish, a quantitative study was performed in order to take away the increment of heart rate due to the water warming, which cannot entirely be suppressed during compression. The specific effects of "per se" pressure have been identified as a bradycardia above 24.5 degrees C and as a tachycardia below this temperature. The use of atropine and propranol, which abolish extrinsic heart control, shows that pressure acts both through extrinsic mechanisms of heart control and through a direct action on autonomic cardiac cells. This effect of "per se" pressure on cardiac cells is interpreted according to chemical kinetics laws. The hypothesis that pressure acts at the molecular level, possibly by the means of reversible structure modifications of some molecules, is suggested.

Animals↗
PubMed1995

Second derivative spectroscopy of enolase at high hydrostatic pressure: an approach to the study of macromolecular interactions.

Second derivative spectroscopy in the ultraviolet region of proteins has been used to study the polarity of the regions surrounding tyrosine residues. We show here that it can also be a tool to study the degree to which proteins associate and that it can be effectively combined with hydrostatic pressure in order to evaluate equilibrium dissociation constants and reaction volumes. Hydrostatic pressure causes yeast enolase to dissociate. Clear changes in the second derivative spectra of enolase were observed as pressure was increased. At enolase concentrations of about 20 microM, the midpoint of the transition is about 1800 bar. All aspects of the transition are reversible up to 2700 bar. It is likely that the transition observed is the result of enolase dimers dissociating into monomers. The second derivative spectra indicate that one or more tyrosine residues is in an unusually polar environment in the dimer, an environment that is less polar in the monomer. Three tyrosines (6, 11, 130) are near the dimer interface. Tyrosines 6 and 11 are pointing into the water-filled crevice between the subunits and are close to several immobilized waters. All three are close to a network of intersubunit salt bridges and hydrogen bonds. We believe that the average tyrosine polarity in the dimer reflects the exposure of these tyrosines to immobilized water and the fixed dipole of the salt bridge. The water in the crevice between the subunits should be more mobile in the monomer; the salt bridge does not exist in the monomer.(ABSTRACT TRUNCATED AT 250 WORDS)

Hydrostatic Pressure↗
PubMed1995

Heterogeneity in molecular recognition by restriction endonucleases: osmotic and hydrostatic pressure effects on BamHI, Pvu II, and EcoRV specificity.

The cleavage specificity of the Pvu II and BamHI restriction endonucleases is found to be dramatically reduced at elevated osmotic pressure. Relaxation in specificity of these otherwise highly accurate and specific enzymes, previously termed "star activity," is uniquely correlated with osmotic pressure between 0 and 100 atmospheres. No other colligative solvent property exhibits a uniform correlation with star activity for all of the compounds tested. Application of hydrostatic pressure counteracts the effects of osmotic pressure and restores the natural selectivity of the enzymes for their canonical recognition sequences. These results indicate that water solvation plays an important role in the site-specific recognition of DNA by many restriction enzymes. Osmotic pressure did not induce an analogous effect on the specificity of the EcoRV endonuclease, implying that selective hydration effects do not participate in DNA recognition in this system. Hydrostatic pressure was found to have little effect on the star activity induced by changes in ionic strength, pH, or divalent cation, suggesting that distinct mechanisms may exist for these observed alterations in specificity. Recent evidence has indicated that BamHI and EcoRI share similar structural motifs, while Pvu II and EcoRV belong to a different structural family. Evidently, the use of hydration water to assist in site-specific recognition is a motif neither limited to nor defined by structural families.

DNA↗