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Results for “Hydrostatic Pressure”

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PubMed1978

Comparison of polymer, glucose, and hydrostatic pressure induced ultrafiltration in a hollow fiber dialyzer: effects on convective solute transport.

Ultrafiltration induced by (1) poly(sodium acrylate), (2) glucose, and (3) hydrostatic pressure was studied in a hollow fiber dialyzer. Poly(sodium acrylate) added to dialysate induced large amounts of ultrafiltration without crossing the dialyzer membrane. Sodium ions of the acrylate polymer were osmotically active but were held in dialysate by the impermeant anionic polymer. The hydrostatic pressure equivalent of osmotic pressure induced primarily by the sodium ions approximated that predicted for a completely impermeant molecule. The apparent (net) sieving coefficients for vitamin B12 observed during polymer and hydrostatic ultrafiltration studies were both significantly higher than that observed during glucose ultrafiltration but did not differ from each other. These studies suggest that sodium salts of polyanions can provide an osmotic driving force to yield large amounts of ultrafiltration in dialysis systems and yet not cross the membrane. The studies also suggest that relatively less efficient convective transport with glucose as compared to hydrostatic pressure is neither a membrane phenomenon nor a characteristic of all osmotic pressure induced ultrafiltration. Relatively low effective solute sieving appears to be associated with osmotic induced ultrafiltration with a permeant solute. Under such conditions it is proposed that molecular interaction within the membrane impairs convective transport.

Acrylic Resins↗

PubMed1998

Subunit interaction of vacuolar H+-pyrophosphatase as determined by high hydrostatic pressure.

Vacuolar H+-pyrophosphatase (H+-PPase) from etiolated hypocotyls of mung bean (Vigna radiata L.) is a homodimer with a molecular mass of 145 kDa. The vacuolar H+-PPase was subjected to high hydrostatic pressure to investigate its structure and function. The inhibition of H+-PPase activity by high hydrostatic pressure has a pressure-, time- and protein-concentration-dependent manner. The Vmax value of vacuolar H+-PPase was dramatically decreased by pressurization from 293.9 to 70.2 micromol of PPi (pyrophosphate) consumed/h per mg of protein, while the Km value decreased from 0.35 to 0.08 mM, implying that the pressure treatment increased the affinity of PPi to vacuolar H+-PPase but decreased its hydrolysis. The physiological substrate and its analogues enhance high pressure inhibition of vacuolar H+-PPase. The HPLC profile reveals high pressure treatment of H+-PPase provokes the subunit dissociation from an active into inactive form. High hydrostatic pressure also induces the conformational change of vacuolar H+-PPase as determined by spectroscopic techniques. Our results indicate the importance of protein-protein interaction for this novel proton-translocating enzyme. Working models are proposed to interpret the pressure inactivation of vacuolar H+-PPase. We also suggest that association of identical subunits of vacuolar H+-PPase is not random but proceeds in a specific manner.

Cell Membrane↗

PubMed1985

Hydrostatic pressure and adrenergic drugs (agonists and antagonists): effects and interactions in fish.

Cardiac responses to alpha and beta adrenergic drugs have been studied in the eel firstly at atmospheric pressure (1 ATA), secondly at 101 ATA per se hydrostatic pressure. The exposure of treated eels for 1 hr at 101 ATA cancels the propranolol effect, increases the clonidine effect and induces paradoxal effects (reversal effect) of isoproterenol, phentolamine and phenylephrine. The highly significant interaction (P less than 0.001) between drugs and HP at 101 ATA is discussed and interpreted as effects of hydrostatic pressure inducing a change in receptor function.

Animals↗

PubMed2000

Combination of hydrostatic pressure and lacticin 3147 causes increased killing of Staphylococcus and Listeria.

The use of hydrostatic pressure and lacticin 3147 treatments were evaluated in milk and whey with a view to combining both treatments for improving the quality of minimally processed dairy foods. The system was evaluated using two foodborne pathogens: Staphylococcus aureus ATCC6538 and Listeria innocua DPC1770. Trials against Staph. aureus ATCC6538 were performed using concentrated lacticin 3147 prepared from culture supernatant. The results demonstrated a more than additive effect when both treatments were used in combination. For example, the combination of 250 MPa (2.2 log reduction) and lacticin 3147 (1 log reduction) resulted in more than 6 logs of kill. Similar results were obtained when a foodgrade powdered form of lacticin 3147 (developed from a spray dried fermentatation of reconstituted demineralized whey powder) was evaluated for the inactivation of L. innocua DPC1770. Furthermore, it was observed that treatment of lacticin 3147 preparations with pressures greater than 400 MPa yielded an increase in bacteriocin activity (equivalent to a doubling of activity). These results indicate that a combination of high pressure and lacticin 3147 may be suitable for improving the quality of minimally processed foods at lower hydrostatic pressure levels.

Animals↗

PubMed1986

Role of a hydrostatic pressure gradient in the formation of early ischemic brain edema.

We studied whether a hydrostatic pressure gradient between arterial blood and brain tissue plays a role in the formation of early ischemic cerebral edema after middle cerebral artery (MCA) occlusion in cats. Tissue pressure, regional CBF, and water content were measured from the cortex in the core and the peripheral zone of brain normally perfused by the MCA. Intraluminal arterial pressure was altered at intervals by inflation of an aortic balloon to vary the blood-tissue pressure gradient in the ischemic zone. Brain water content in the ischemic core, where flow fell to 5.5 ml/100 g/min, increased within 1 h of occlusion. After occlusion tissue pressure rose from 7.95 +/- 0.72 mm Hg at 1 h to 13.16 +/- 1.13 mm Hg at 3 h. When intraluminal pressure was increased, water content increased further, but only at 1 h after occlusion. In the periphery where flow was 18.9 ml/100 g/min during normotension, neither water content nor tissue pressure rose within 3 h of occlusion. Increased intraluminal pressure was accompanied by increased water content only at 3 h. This study indicates that a hydrostatic pressure gradient is an important element in the development of ischemic brain edema, exerting its major effect during the initial phase of the edema process.

Animals↗

PubMed1990

Effects of hydrostatic pressure and inert gases on platelet aggregation in vitro.

A novel cuvette was used to subject citrated platelet-rich plasma (PRP) to high hydrostatic pressure with negligible contamination by He (used for compression of the apparatus). Aggregation was induced at pressure by ADP and quantified turbidimetrically. The maximum degree of aggregation (MDA) was reduced from a control level of 82.2 to 53.6% by exposure to 101 ATA. Because decompression bubbles did not form, aggregation was also measured immediately after a compression cycle. After exposure to 101 ATA hydrostatic pressure, platelets responded normally to ADP at 1 ATA. In a matching apparatus, PRP was equilibrated with high partial pressures of inert gases. Normal physiological plasma Po2 and pH were maintained during equilibration. N2O (5 ATA) reduced the MDA from 86.5 (control) to 58.1%. N2 (51 ATA) reduced the MDA from 74.7 (control) to 51.6%, and 101 ATA Pn2 reduced the MDA from 78.0 (control) to 32.3%. He (100 ATA) reduced the MDA from 83.6 to 38.6%. It was concluded that platelet aggregation was relatively sensitive to hydrostatic pressure and less sensitive to inert gases than predicted from their anesthetic potency ratios.

Adenosine Diphosphate↗

PubMed1999

The effects of sustained hydrostatic pressure on select bladder smooth muscle cell functions.

PURPOSE: Normal bladder development is believed to depend on the active work of the bladder for storing and expelling urine. When high urinary diversion is performed in infants and the bladder no longer undergoes normal filling, bladder development may be altered, ultimately resulting in bladder dysfunction. To help better understand this relationship of bladder function with growth at the cellular level we developed a novel laboratory method for applying hydrostatic pressure to cell cultures, and we characterized the response of neonatal bladder smooth muscle cells to physiological levels of sustained hydrostatic pressure. MATERIALS AND METHODS: Neonatal ovine smooth muscle cells staining positive for desmin and alpha-smooth muscle actin were exposed to pressures of 0.3 (controls), 2, 4, 6 and 8.5 cm. water for 1, 3, 5 and 7 days. At the end of the experiments the cells were fixed, stained and counted. Mitogenic activity of the supernatant media from bladder smooth muscle cells exposed to 8.5 cm. water for 5 days (conditioned media) was tested before and after treatments of heating, freezing, passing through a heparin-sepharose affinity chromatography column or after the addition of suramin, a nonspecific growth factor inhibitor. Statistical analysis was performed using Student's t test with p <0.05 considered statistically significant. RESULTS: Exposure of bladder smooth muscle cells to sustained hydrostatic pressures of 4, 6 and 8.5 cm. water resulted in increased cell proliferation. Differences became statistically significant (p <0.05) by day 5. Also, conditioned media contained mitogenic activity that was ablated by heating, freezing, passage through a heparin-sepharose affinity chromatography column or with the addition of suramin. CONCLUSIONS: We have demonstrated a proliferative response of neonatal bladder smooth muscle after exposure to physiological levels of sustained hydrostatic pressure. This response is partially due to 1 or more transferable mitogenic factors. These data support the hypothesis that pressure associated with bladder filling is an important stimulus for detrusor development.

Animals↗

PubMed1996

Biological alterations of rat podocytes cultured under basolateral hydrostatic pressure.

In vivo, glomerular visceral epithelial cells (GVEC), or podocytes, are morphologically highly differentiated cells which are in close contact with adjacent cells by complex interdigitating foot processes. In vitro, the dedifferentiated appearance of podocytes hampers investigations on podocyte structure and function. Cultured podocytes resemble simple epithelium in several ways with apical tight junctions and absence of foot processes. The morphological resemblances between GVEC early in embryonic development, in proteinuric diseases and in cultured cells are striking, but the mechanisms involved in these (de)differentiation processes are poorly understood. A common feature of GVEC in these various states of dedifferentiation is their altered exposure to or even total lack of hydrostatic pressure, suggesting that this may be one of the parameters involved in GVEC differentiation. In this study we investigated whether basolateral hydrostatic pressure could affect GVEC biology in vitro. We therefore exposed cultured GVEC grown on porous supports to basolateral hydrostatic pressure and investigated morphology with scanning and transmission electron microscopy, expression of specific podocyte markers and their biological responses to a model stimulus, the cytokine IFN-gamma. Morphologically, monolayers of pressurized GVEC contained large regions of whirl-like, raised cell formations. Individual cells in these formations had a rounded morphology and pore-like indentations between adjacent cells were observed. Cell-cell contacts were often found more basally and intercellular spaces were widened. Moreover, protein expression of pressurized monolayers was altered, as demonstrated by regions of cells with decreased keratin expression. Finally, upon exposure to the model stimulus IFN-gamma, the pressurized as compared to the control GVEC demonstrated a 3-fold increased expression of MHC class II and a strongly decreased sensitivity to the toxic effects of IFN-gamma. In conclusion, we found several indications that hydrostatic pressure can affect podocyte biology in vitro and similar mechanisms may account for podocyte biology in vivo. The strikingly altered morphology and biology of pressurized GVEC suggest that this culture system can be quite relevant for future studies with cultured GVEC.

Animals↗

PubMed1989

[Effects of hydrostatic pressure on proliferation and expression of the differentiated phenotype of cultured chondrocytes derived from rabbit craniofacial complex].

To investigate the effects of mechanical forces on the growth of cartilages of the craniofacial complex, chondrocytes isolated from nasal septal cartilage (NSC), spheno-occipital synchondrosis (SOS), and mandibular condylar cartilage (MCC) were cultured and exposed to hydrostatic pressure in vitro. Following results were obtained. 1. By application of the pressure, DNA synthesis increased in NSC and SOS (150% of control by 50 g/cm2, 1 min), but not so much in MCC in the medium containing 10% fetal calf serum (FCS). On the other hand, GAG synthesis increased in SOS (140% of control by 50 g/cm2, 2 min.) and MCC (160% of control by 100 g/cm2, 5 min), but decreased slightly in NSC in the same condition. The stimulations in DNA and GAG syntheses reached maxima 20-22 hr and 24-27 hr respectively after the application of the pressure. These stimulatory effects of the hydrostatic pressure on DNA and GAG syntheses could be found when cultured chondrocytes were in a multilayer stage. 2. It has been reported that addition of FCS at a concentration of 10% increases DNA synthesis in these chondrocytes preincubated in medium containing 0.3% FCS. The application of the pressure 2 min before the addition of 10% FCS potentiated the increase by FCS of DNA synthesis in NSC (130% of control), but not so much that in SOS and little that in MCC. 3. When the pressure was applied on these cells 2 min before addition of parathyroid hormone (PTH; 10(-7) M), this mechanical stimulation potentiated the increase by PTH of GAG synthesis in MCC (130% of control), but not so much that in SOS, and little that in NSC. Moreover, the intracellular cAMP level which was increased just after the addition of PTH also became higher by the application of the pressure. 4. These potentiation by the hydrostatic pressure in the cases of the addition of FCS and PTH diminished when intervals between application of pressure and addition of these factors were more than 10 min. These findings suggested that mechanical forces have influences on the growth of cartilages in craniofacial complex and that these effects are mediated by factors contained in serum.

Animals↗

PubMedDate not supplied

Electrical and morphological changes of human erythrocytes under high hydrostatic pressure followed by dielectric spectroscopy.

The electrical and morphological properties of human erythrocytes under high hydrostatic pressure up to 500 MPa have been studied by dielectric spectroscopy. The pressure-induced changes in the dielectric behavior of erythrocyte suspensions indicate that hydrostatic pressure causes the change in cell shape from discoidal to spherical and hemolysis at 200-300 MPa, the formation of buds and spicular processes followed by vesiculation at 300-400 MPa, and the increase in the membrane capacitance at 400-500 MPa.

Cell Membrane↗

PubMed2003

Properties of diamond under hydrostatic pressures up to 140 GPa.

Diamond is the archetypal covalent material. Each atom in an sp(3) configuration is bonded to four nearest neighbours. Because of its remarkable properties, diamond has been extensively studied. And yet our knowledge of the properties of diamond under very high pressure is still incomplete. Although diamond is known to be the preferred allotrope of carbon at high pressure, the possibility of producing under pressure high-density polymorphs of diamond, including metallic forms, has been discussed. Structural changes have already been reported in diamond under non-hydrostatic pressures around 150 GPa and large deformation. However, measurements of the properties of diamond under hydrostatic pressure have been limited to below 40 GPa. Here, we report accurate measurements of the volume and of the optical phonon frequency of diamond under hydrostatic pressure up to 140 GPa. We show that diamond is more compressible than currently expected. By combining the volume and the frequency pressure shifts, we deduce that diamond remains very stable under pressure: it is a Gruneisen solid up to at least 140 GPa, and the covalent bond is even slightly strengthened under pressure. Finally, the optical phonon frequency versus pressure is calibrated here to be used as a pressure gauge for diamond anvil cell studies in the multi-megabar range.

Crystallography, X-Ray↗

PubMed2006

Chondrocyte response to cyclic hydrostatic pressure in alginate versus pellet culture.

Cells are often cultured at high density (e.g., confluent monolayer and as pellets) to promote chondrogenic differentiation and to maintain the chondrocyte phenotype. They are also frequently suspended in hydrogels such as agarose or alginate for the same purposes. These culture techniques differ markedly with respect to frequency of direct contact between cells and overall intercellular spacing. Because these factors may significantly affect mechanotransduction, the purpose of this study was to determine if the response of articular chondrocytes to cyclic hydrostatic pressure would depend on the culture condition. Primary articular chondrocytes from young and mature pigs were cultured either as pellets or suspended in alginate beads. Both groups were exposed to dynamic hydrostatic pressure (4 MPa, 1 Hz, 5400 cycles per day) for 7 days. Cell proliferation was unaffected by pressure, but pressurized chondrocytes in pellet culture had significantly greater sGAG content and incorporated [3H]proline at a higher rate than nonpressurized controls. Electron microscopy revealed a fibrous extracellular matrix (ECM) surrounding pellets, but not cells in alginate. In addition, expression of Connexin 43 (Cx43) mRNA was slightly lower in alginate than in pellet cultures and was not significantly altered by loading. Thus, metabolic response of chondrocytes to dynamic hydrostatic pressure was affected by culture technique; chondrocytes cultured as pellets exhibited the classical anabolic response to dynamic hydrostatic pressure, but those in alginate did not. Although cell-ECM interaction could be important, the differential response is not likely attributable to differential expression of Cx43 mRNA.

Alginates↗

PubMed1989

Role of renal interstitial hydrostatic pressure in the pressure diuresis response.

The present study examines the role of renal interstitial hydrostatic pressure (RIHP) in the pressure-diuretic and -natriuretic response. The relationships between RIHP, sodium excretion, and renal perfusion pressure (RPP) were determined in antidiuretic and volume-expanded (VE) rats with an intact or decapsulated kidney. RIHP was measured by use of the implanted capsule technique. RIHP increased significantly from 7.5 +/- 0.8 to 12.0 +/- 1.4 mmHg in VE animals and from 3.3 +/- 0.4 to 5.2 +/- 0.7 mmHg in antidiuretic rats after RPP was varied from 100 to 150 mmHg. The pressure-natriuretic response of the antidiuretic rats was blunted compared with that observed in the VE rats. Decapsulation of the kidney in VE rats lowered RIHP and reduced, but did not eliminate, the pressure-natriuretic response. To determine whether this residual response was related to changes in interstitial pressure in the medulla, cortical (CIHP) and medullary interstitial hydrostatic pressures (MIHP) were simultaneously measured in VE rats with an intact or decapsulated kidney. In control rats CIHP and MIHP were similar at all levels of RPP studied. In rats with the renal capsule removed MIHP was higher than CIHP and rose significantly from 6.7 +/- 0.8 to 9.2 +/- 0.8 mmHg when RPP was varied from 100 to 150 mmHg. These results indicate that pressure diuresis and natriuresis is accompanied by changes in RIHP and the response is modulated by the basal level of RIHP. These findings suggest that changes in MIHP may serve as an intrarenal signal for this response.

Animals↗

PubMed2000

Compressibility and polygonization of single-walled carbon nanotubes under hydrostatic pressure

Single-walled carbon nanotubes show linear elasticity under hydrostatic pressure up to 1.5 GPa at room temperature. The volume compressibility, measured by in situ synchrotron x-ray diffraction, has been determined to be 0.024 GPa (-1). Theoretical calculations suggest that single-walled carbon nanotubes are polygonized when they form bundles of hexagonal close-packed structure and the intertubular gap is smaller than the equilibrium spacing of graphite (002) (d = 3.35 A). It has also been determined that the deformation of the trigonal nanotube lattice under hydrostatic pressure is reversible up to 4 GPa, beyond which the nanotube lattice is destroyed.

Journal Article↗

PubMed2007

A new experimental system for the extended application of cyclic hydrostatic pressure to cell culture.

Mechanical forces have been shown to be important stimuli for the determination and maintenance of cellular phenotype and function. Many cells are constantly exposed in vivo to cyclic pressure, shear stress, and/or strain. Therefore, the ability to study the effects of these stimuli in vitro is important for understanding how they contribute to both normal and pathologic states. While there exist commercial as well as custom-built devices for the extended application of cyclic strain and shear stress, very few cyclic pressure systems have been reported to apply stimulation longer than 48 h. However, pertinent responses of cells to mechanical stimulation may occur later than this. To address this limitation, we have designed a new cyclic hydrostatic pressure system based upon the following design variables: minimal size, stability of pressure and humidity, maximal accessibility, and versatility. Computational fluid dynamics (CFD) was utilized to predict the pressure and potential shear stress within the chamber during the first half of a 1.0 Hz duty cycle. To biologically validate our system, we tested the response of bone marrow progenitor cells (BMPCs) from Sprague Dawley rats to a cyclic pressure stimulation of 120/80 mm Hg, 1.0 Hz for 7 days. Cellular morphology was measured using Scion Image, and cellular proliferation was measured by counting nuclei in ten fields of view. CFD results showed a constant pressure across the length of the chamber and no shear stress developed at the base of the chamber where the cells are cultured. BMPCs from Sprague Dawley rats demonstrated a significant change in morphology versus controls by reducing their size and adopting a more rounded morphology. Furthermore, these cells increased their proliferation under cyclic hydrostatic pressure. We have demonstrated that our system imparts a single mechanical stimulus of cyclic hydrostatic pressure and is capable of at least 7 days of continuous operation without affecting cellular viability. Furthermore, we have shown for the first time that BMPCs respond to cyclic hydrostatic pressure by alterations in morphology and increased proliferation.

Animals↗

PubMed2002

Renal interstitial hydrostatic pressure and natriuretic responses to volume expansion in pregnant rats.

During normal pregnancy, a gradual plasma volume expansion (VE) occurs and reaches a maximum level at late term. Pressure natriuresis and renal interstitial hydrostatic pressure (RIHP) responses are attenuated in pregnant rats. Also, basal RIHP is lower in pregnant rats, suggesting an increase in renal interstitial compliance during pregnancy. This adaptation may contribute to the increase in plasma volume that is required for a normal pregnancy, because increases in RIHP have been consistently shown to produce natriuresis and diuresis. Acute saline VE (5% body wt/30 min) has been shown to increase RIHP in normal nonpregnant rats. Therefore, the objective of this study was to determine RIHP, natriuretic, and diuretic responses to VE in nonpregnant (n = 7), midterm pregnant (n = 8), and late-term pregnant (n = 8) Sprague-Dawley rats. Although VE significantly increased RIHP, fractional excretion of sodium (FE(Na)), and urine flow rate (V) in all groups, DeltaRIHP was highest for nonpregnant (3.0 +/- 0.3 mmHg) compared with midterm pregnant (1.6 +/- 0.1 mmHg; P < 0.05 vs. nonpregnant) and late-term pregnant rats (1.2 +/- 0.1 mmHg; P < 0.05 vs. both midterm pregnant and nonpregnant rats). DeltaFE(Na) and DeltaV were similar in all groups: 5.8 +/- 1.0% and 231 +/- 27 microl/min for nonpregnant, 6.8 +/- 1.3% and 173 +/- 16 microl/min for midterm pregnant, and 7.6 +/- 1.2% and 203 +/- 10 microl/min for late-term pregnant rats, respectively. In conclusion, basal RIHP and the increase in RIHP during VE were attenuated during pregnancy; however, the natriuretic and diuretic responses to VE remain intact during the course of pregnancy.

Absorption↗

PubMed2005

Hydrostatic pressure affects the conformational equilibrium of Salmonella typhimurium tryptophan synthase.

The effect of hydrostatic pressure on the tryptophan (Trp) synthase alpha2beta2 complex from Salmonella typhimurium has been investigated. Trp synthase has been shown previously to exhibit low-activity (open) and high-activity (closed) conformations. The equilibrium between the open and closed conformations of Trp synthase has been found to be affected by a wide range of variables, including alpha-subunit ligands, monovalent cations, organic solvents, pH, and temperature. The absorption spectrum of the Trp synthase-L-Ser complex shows an increase in absorption of the 423 nm band of the external aldimine, which is a characteristic of the open conformation, as hydrostatic pressure is increased from 1 to 2000 bar. The deltaV(o) and K(o) for the equilibrium between the closed and open conformations of the Trp synthase-L-Ser complex are -126 mL/mol and 0.12 for the Na+ form and -171 mL/mol and 2.3 x 10(-4) for the NH4+ form. When the Trp synthase-L-Ser complex is subjected to pressure jumps of 100-400 bar, relaxations are observed, exhibiting an increase in fluorescence emission at wavelengths greater than 455 nm, with 405 nm excitation. The relaxation to the new equilibrium position requires two exponentials to fit the data in the presence of 0.1 M Na+ and three exponentials to obtain a reasonable fit in the absence of cations and with 0.1 M NH4+. Fluorescence emission at 325 nm, with excitation at 280 nm, also increases when the Trp synthase-L-Ser complex is subjected to pressure jump. These data demonstrate that the open conformation of Trp synthase is favored by higher pressure. Thus, the open conformation has a smaller apparent net system volume than the closed conformation. We estimate that there are 35-47 more waters in the solvation shell of the open conformation than in that of the closed conformation.

Bacterial Proteins↗

PubMed1997

Ability of high hydrostatic pressure treated plasmids and cells of Escherichia coli to genetically transform.

The exposure of plasmid pUC18 and pBR322 DNA to high hydrostatic pressure increased the ability of plasmids to transform competent Escherichia coli cells. For pUC18 plasmid, a pressure of 400 MPa, and for pBR322, a pressure of 200 MPa was found to provide the highest transformation efficiency. The DNA duplexes of the two plasmids were found to be the most stable for melting conditions at these pressures. At pressures higher than these, both the stability of the duplex DNA and the transformation efficiency were affected. The stabilizing effect of high hydrostatic pressure on the hydrogen bond may be responsible for the observed increase in transformation efficiency of the pressure-exposed plasmid DNA. The possibility of pressure-induced changes in the structure and conformation of DNA was studied using various techniques. In agarose gel electrophoresis, pressure-treated plasmids (pUC18 at 400 MPa and pBR322 at 200 MPa) consistently showed visibly distinct higher mobility compared to untreated plasmids. Pressure-treated pUC18 as well as pBR322 DNA showed significant reduction in ethidium bromide binding as is evident from the reduced intensity of fluorescence of the dye bound pressure-treated DNA. Spectroscopic studies using circular dichroism and Fourier transform infrared (FTIR) spectroscopy also showed significant differences in the absorption profiles of pressure-treated plasmids as compared to an untreated control. These studies revealed that the pressure-induced changes in the conformation of these DNAs may be responsible for the observed increase in the transformation ability of the plasmids. On the other hand, the exposure of competent cells of E. coli to a high hydrostatic pressure of 50 MPa not only reduced their colony-forming ability but also drastically reduced their ability to take up plasmid DNA.

Journal Article↗