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The influence of increased tubular hydrostatic pressure on renal function.

The effect of tubular obstruction on renal function has been understood poorly at the tubular level and from the clinical standpoint. In our review the evidence for a direct influence of hydrostatic pressure on tubular transport and glomerular filtration is examined. The data generated to date indicate a direct influence of hydrostatic pressure on tubular transport only at the level of the distal convoluted tubule and collecting duct. With respect to glomerular filtration increased tubular pressure reduces the net driving force for filtration and reduces glomerular filtration rate in the absence of a compensatory increase in glomerular hydrostatic pressure. We next review physiological data concerning the mechanism of post-obstructive diuresis. Available information suggests 4 factors that play a significant role in the clinical syndrome of post-obstructive diuresis: 1) medullo-papillary washout, 2) decreased fractional and absolute salt and water reabsorption in the collecting duct, presumably secondary to direct influence of hydrostatic pressure on transport mechanisms, 3) osmotic diuresis secondary to retention of urea and other osmotic solutes during the period of obstruction and 4) prior salt and water administration in the absence of excretion, resulting in extracellular fluid volume expansion.

Diuresis↗

Effects of hydrostatic pressure on sensory discharge in frog semicircular canals.

The effects of endolymphatic and perilymphatic pressure changes on resting and mechanically evoked responses were studied in isolated posterior semicircular canals of the frog. The results demonstrated that ampullar receptors are extremely sensitive to hydrostatic pressure changes (0.25 mm H2O were sufficient to produce distinct changes), being inhibited by endolymphatic pressure increases and facilitated by perilymphatic ones. Intracellular recordings from single afferent axons showed that the effects of hydrostatic pressure result from a modified transmitter release from the synaptic pole of the hair cells. Unlike resting activity, mechanically evoked activity was always depressed in the presence of a hydrostatic pressure. This indicates that the sensitivity of ampullar receptors to mechanical stimuli, i.e. the gain of the conversion process, is maximal when no pressure is present between the inner and the outer fluid. The possible action of hydrostatic pressure on vestibular receptors is discussed.

Afferent Pathways↗

Hydrostatic pressure as an office procedure for congenital nasolacrimal duct obstruction.

BACKGROUND: The estimated incidence of congenital nasolacrimal duct obstruction (CNDO) is 5% to 20%. We examined our success rate of treating CNDO with hydrostatic pressure (the Crigler method) as an office procedure. METHODS: A total of 742 children with CNDO initially were treated noninvasively with hydrostatic pressure. The procedure was safely repeated up to 3 times, with an interval of at least 1 week, if the condition persisted. RESULTS: Success was defined as no epiphora or discharge. The success rate for the entire study group was 45% for children up to 1 year of age. The ducts were opened in 46% (343/742) at the first attempt of hydrostatic pressure, in 35% (42/120) at the second attempt, and in 38% (8/21) at the third attempt. When the maneuver was performed in patients younger than 2 months of age, the success rate of the first attempt was 56%, decreasing to 46% in children 2 to 6 months of age and to 28% older than 6 months of age. CONCLUSIONS: Hydrostatic pressure is an effective way of resolving infant CNDO and shortening the duration of the associated morbidity. The success rate is higher when the procedure is conducted in patients up to 2 months of age. Nevertheless, we recommend this approach for every infant presenting with CNDO at the first office visit, even after 6 months of age. Procedures can be repeated safely and successfully.

Female↗

Hydrostatic pressure promotes the acidification of vacuoles in Saccharomyces cerevisiae.

Application of hydrostatic pressure caused a delay or cessation of cell growth in Saccharomyces cerevisiae. The yeast vacuole is an acidic organelle involved in cellular ion homeostasis and degradation of proteins. Hydrostatic pressure promoted the acidification of the vacuoles in the strain IFO 2347. A pressure of 40 to 60 MPa reduced the vacuolar pH, defined using 6-carboxyfluorescein, from 6.05 to 5.88, while a pressure of 20 MPa did not affect the pH. Similar results were obtained with the strain X2180. Bafilomycin A1, a specific inhibitor of vacuolar H(+)-ATPase (V-H(+)-ATPase), caused a significant alkalization of vacuoles in the strain X2180. The pHs rose to 7.34 and 6.84 at both atmospheric pressure and a pressure of 40 MPa, respectively. Meanwhile, vacuolar accumulation of the weak base quinacrine was increased by a pressure of 40 MPa, suggesting that uptake of the dye was induced by the increased pH gradient across the vacuolar membrane.

Anti-Bacterial Agents↗

Oscillatory shear stress and hydrostatic pressure modulate cell-matrix attachment proteins in cultured endothelial cells.

Endothelial cells (ECs) may behave as hemodynamic sensors, translating mechanical information from the blood flow into biochemical signals, which may then be transmitted to underlying smooth muscle cells. The extracellular matrix (ECM), which provides adherence and integrity for the endothelium, may serve an important signaling function in vascular diseases such as atherogenesis, which has been shown to be promoted by low and oscillating shear stresses. In this study, confluent bovine aortic ECs (BAECs), were exposed to an oscillatory shear stress or to a hydrostatic pressure of 40 mmHg for time periods of 12 to 48 h. Parallel control cultures were maintained in static condition. Although ECs exposed to hydrostatic pressure or to oscillatory flow had a polygonal morphology similar to that of control cultures, these cells possessed more numerous central stress fibers and exhibited a partial loss of peripheral bands of actin, in comparison to static cells. In EC cultures exposed to oscillatory flow or hydrostatic pressure, extracellular fibronectin (Fn) fibrils were more numerous than in static cultures. Concomitantly, a dramatic clustering of alpha 5 beta 1 Fn receptors and of the focal contact-associated proteins vinculin and talin occurred. Laminin (Ln) and collagen type IV formed a network of thin fibrils in static cultures, which condensed into thicker fibers when BAECs were exposed to oscillatory shear stress or hydrostatic pressure. The ECM-associated levels of Fn and Ln were found to be from 1.5- to 5-fold greater in cultures exposed to oscillatory shear stress or pressure for 12 and 48 h, than in static cultures. The changes in the organization and composition of ECM and focal contacts reported here suggest that ECs exposed to oscillatory shear stress or hydrostatic pressure may have different functional characteristics from cells in static culture, even though ECs in either environment exhibit a similar morphology.

Actins↗

Increased drainage volume in intermittent peritoneal dialysis using a two-bag, low hydrostatic pressure drainage system.

Adequate drainage volume (DV) is a prerequisite for long-term success of intermittent peritoneal dialysis (IPD). Ultrafiltration (UF) is basically determined by the interplay of two major forces: a driving osmotic force created by a hyperosmolar dialysate solution versus an opposing force--the intraperitoneal hydrostatic pressure. Conventional drainage procedures (CS) are capable of achieving a fluid volume of 3 L per bag. We found that this intrabag volume corresponds to an intrabag hydrostatic pressure of 60 cm water. Because this drainage system is closed, the intrabag hydrostatic pressure reflects intraperitoneal hydrostatic pressure. A low-pressure system (LPS) was devised to evaluate the possible influence on DV of reducing HP. Our results show that such LPS does indeed significantly improve DV, using a decreased number of exchanges. The residual volume and its glucose concentration (in a 1.5-g/dl exchange) were measured with CS. When followed by an 4.25-g/dl exchange, an 11% reduction in glucose concentration was evident. Thus, the osmotic driving force may also be impaired by incomplete drainage. To obtain maximal DV during IPD, a LPS should be used. This could be achieved by using 2 L dialysate in 3-L bags.

Aged↗

Influence of cyclic hydrostatic pressure on fibrocartilaginous metaplasia of achilles tendon fibroblasts.

The goal of this study was to demonstrate whether cyclically imposed hydrostatic pressure, compressive in nature, could induce fibrocartilaginous metaplasia in a purely tendinous cell source in vitro. The effect of short-duration cyclic hydrostatic pressure on tendon fibroblasts (tenocytes) expanded from rat Achilles tendon was studied. Total RNA was isolated either immediately after loading or 24 h later. The mRNA expression of tendon and cartilage specific markers - Collagen types I and II, Sox9, and Aggrecan was quantified by real-time reverse transcription polymerase chain reaction over multiple biological samples (n=6). For immediately isolated RNA samples, there were statistically significant increases in mRNA expression of Aggrecan and Collagen type II, while Collagen type I significantly decreased. Noticeably, for RNA samples isolated 24 h later, there were further increases in mRNA expression of Aggrecan and Collagen type II, whereas Collagen type I increased roughly three-fold relative to the non-loaded control. These findings support the hypothesis that cyclic hydrostatic pressurization can induce fibrocartilaginous metaplasia in tenocytes by upregulation of cartilaginous gene expression. Also, it was demonstrated that changes in mRNA expression as a result of single 2 h pressurization persist even up to 24 h.

Achilles Tendon↗

Metabolic selectivity and growth of Clostridium thermocellum in continuous culture under elevated hydrostatic pressure.

The continuous culture of Clostridium thermocellum, a thermophilic bacterium capable of producing ethanol from cellulosic material, is demonstrated at elevated hydrostatic pressure (7.0 MPa, 17.3 MPa) and compared with cultures at atmospheric pressure. A commercial limitation of ethanol production by C. thermocellum is low ethanol yield due to the formation of organic acids (acetate, lactate). At elevated hydrostatic pressure, ethanol:acetate (E/A) ratios increased >10(2) relative to atmospheric pressure. Cell growth was inhibited by approximately 40% and 60% for incubations at 7.0 MPa and 17.3 MPa, respectively, relative to continuous culture at atmospheric pressure. A decrease in the theoretical maximum growth yield and an increase in the maintenance coefficient indicated that more cellobiose and ATP are channeled towards maintaining cellular function in pressurized cultures. Shifts in product selectivity toward ethanol are consistent with previous observations of hydrostatic pressure effects in batch cultures. The results are partially attributed to the increasing concentration of dissolved product gases (H2, CO2) with increasing pressure; and they highlight the utility of continuous culture experiments for the quantification of the complex role of dissolved gas and pressure effects on metabolic activity.

Acetic Acid↗

Hsp70 accumulation in chondrocytic cells exposed to high continuous hydrostatic pressure coincides with mRNA stabilization rather than transcriptional activation.

In response to various stress stimuli, heat shock genes are induced to express heat shock proteins (Hsps). Previous studies have revealed that expression of heat shock genes is regulated both at transcriptional and posttranscriptional level, and the rapid transcriptional induction of heat shock genes involves activation of the specific transcription factor, heat shock factor 1 (HSF1). Furthermore, the transcriptional induction can vary in intensity and kinetics in a signal- and cell-type-dependent manner. In this study, we demonstrate that mechanical loading in the form of hydrostatic pressure increases heat shock gene expression in human chondrocyte-like cells. The response to continuous high hydrostatic pressure was characterized by elevated mRNA and protein levels of Hsp70, without activation of HSF1 and transcriptional induction of hsp70 gene. The increased expression of Hsp70 was mediated through stabilization of hsp70 mRNA molecules. Interestingly, in contrast to static pressurization, cyclic hydrostatic loading did not result in the induction of heat shock genes. Our findings show that hsp70 gene expression is regulated posttranscriptionally without transcriptional induction in chondrocyte-like cells upon exposure to high continuous hydrostatic pressure. We suggest that the posttranscriptional regulation in the form of hsp70 mRNA stabilization provides an additional mode of heat shock gene regulation that is likely to be of significant importance in certain forms of stress.

Cartilage↗

Effect of extracorporal high hydrostatic pressure on tumor cell adherence and viability.

In orthopedic surgery, reconstruction of bone segments afflicted with cancer is done in various ways, including devitalization of the bone or replacement of the bone by artificial bone constructs. To devitalize bone cells, extracorporal irradiation or autoclaving is used although both methods have substantial disadvantages. We now introduce the technique of extracorporal high hydrostatic pressure (HHP) treatment to disintegrate tumor cells in suspension or in their adherent state. The effect of HHP on cell viability, adherence and morphology of four different tumor cell lines (fibrosarcoma HT-1080, osteosarcoma SAOS-2, ovarian cancer OV-MZ-6, breast cancer MCF-7) was investigated. For this, adherently growing (with fibronectin serving as the growth-promoting substrate) or suspended tumor cells were placed into a test vial which was transferred into the pressure chamber of a high hydrostatic pressure device. After pressure treatment, the pressure was relaxed to atmospheric pressure and subsequently cell viability, adherence and morphology assessed. High hydrostatic pressure as high as 350 MPa (10 min, 37 degrees C) did not detach the tumor cells from the fibronectin-coated surface although at these conditions all of the four cell lines tested were irreversibly damaged. Adherently growing tumor cells were considerably more sensitive to HHP than tumor cells detached from the surface and treated by HHP in suspension. HHP-treated tumor cells showed drastic morphological changes, evident by cell membrane ruffling and bleb formation. At 150 MPa adherently growing or suspended tumor cells are irreversibly damaged by short-term treatment with HHP. In another investigation, we experienced that treatment of freshly excised bones or tendons by HHP has no adverse effect on their stability or biomechanical properties. Therefore, we anticipate that in orthopedic surgery HHP could be used as a new gentle way of treating resected cancer-afflicted bones or tendons to inactivate tumor cells before autologous reimplantation.

Bone and Bones↗

High hydrostatic pressure inactivation of Salmonella typhimurium: effects of pressure, duration, pH and temperature studied by analysis of variance.

High hydrostatic pressure treatments are regarded as possible alternative methods for food preservation. One of the primary considerations for industrial applications is the ability of these methods to eradicate pathogenic microorganisms. This study subjected S. typhimurium suspensions, first in a phosphate buffer (pH 7.0) and then in a citrate phosphate buffer (pH 5.6), to high hydrostatic pressure treatments relative to the following variables: pressure (200-400 MPa), duration (3, 10 and 20 min), temperature (4, 20 and 40 degrees C) and the pH of the suspension medium (5.6 and 7.0). An optimal design of 40 runs was obtained using the Fedorov algorithm, and responses were studied by analysis of variance in terms of cell survival on plate count agar. Efficiency was determined by Log10 comparisons of the numbers of live cells before and after treatment. A statistically significant relationship was found between the four variables considered (pressure, pH, duration and temperature), their interactions (duration x pressure, pH x temperature, pH x pressure) and the inactivation of S. typhimurium. R-squared statistical analysis indicated that the linear model used accounted for more than 98% of the variability in the inactivation of S. typhimurium.

Algorithms↗

Time-dependent effects of intermittent hydrostatic pressure on articular chondrocyte type II collagen and aggrecan mRNA expression.

The normal loading of joints during daily activities causes the articular cartilage to be exposed to high levels of intermittent hydrostatic pressure. This study quantified effects of intermittent hydrostatic pressure on expression of mRNA for important extracellular matrix constituents. Normal adult bovine articular chondrocytes were isolated and tested in primary culture, either as high-density monolayers or formed aggregates. Loaded cells were exposed to 10 MPa of intermittent hydrostatic pressure at a frequency of 1 Hz for periods of 2, 4, 8, 12, and 24 hrs. Other cells were intermittently loaded for a period of 4 hrs per day for 4 days. Semiquantitative reverse transcription polymerase chain reaction assays were used to assess mRNA signal levels for collagen types II and I and aggrecan. The results showed that type II collagen mRNA signal levels exhibited a biphasic pattern, with an initial increase of approximately five-fold at 4 and 8 hrs that subsequently decreased by 24 hrs. In contrast, aggrecan mRNA signal increased progressively up to three-fold throughout the loading period. Changing the loading profile to 4 hrs per day for 4 days increased the mRNA signal levels for type II collagen nine-fold and for aggrecan twenty-fold when compared to unloaded cultures. These data suggest that specific mechanical loading protocols may be required to optimally promote repair and regeneration of diseased joints.

Aggrecans↗

Hydrostatic pressure influences histone mRNA.

Exposure of HeLa S3 cells to high hydrostatic pressure (6.89 x 10(3) to 6.89 x 10(4) kPa: 1000 to 10,000 lbfin-2) reduced core and H1 histone mRNA levels as determined by hybridization to specific histone DNA probes. At 4.14 x 10(4) kPa for 10 min core histone and H1 histone mRNA levels were reduced 32-38% and 56%, respectively. At 30 min postdecompression core mRNA levels returned to atmospheric control levels while H1 histone mRNA levels continued to be suppressed. Levels of macromolecular synthesis were monitored under hydrostatic pressure with radioactive precursors of RNA, DNA and protein. Macromolecular synthesis was shown to be suppressed in a dose-dependent manner with increasing magnitude and duration of pressure. To determine the influence of pressure on histone mRNA stability, actinomycin D (10 micrograms ml-1) was used to block RNA synthesis. Relative amounts of H4 and H1 mRNA were determined at atmospheric pressure and following treatment with actinomycin D (10 micrograms ml-1), pressure (4.14 x 10(4) kPa) and a combination of pressure and actinomycin D. This study shows that a synthesis component and a stability component are involved in the pressure-induced reduction of core histone mRNA. At 4.14 x 10(4) kPa for 15 min, there was a 42% reduction in core histone mRNA of which approximately one third was due a suppression of transcription and two thirds to a loss of mRNA stability. The pressure-induced reduction in histone mRNA is attributed to the instability of endogenous histone mRNA and a reduction in transcription/processing of new histone mRNA.

HeLa Cells↗

Elasto-optics in double-coated optical fibers induced by axial strain and hydrostatic pressure.

Stresses, microbending loss, and refractive-index changes induced simultaneously by axial strain and hydrostatic pressure in double-coated optical fibers are analyzed. The lateral pressure and normal stresses in the optical fiber, primary coating, and secondary coating are derived. Also presented are the microbending loss and refractive-index changes in the glass fiber. The normal stresses are affected by axial strain, hydrostatic pressure, material properties, and thickness of the primary and secondary coatings. It is found that microbending loss decreases with increasing thickness, the Young's modulus, and the Poisson's ratio of the secondary coating but increases with the increasing Young's modulus and Poisson's ratio of the primary coating. Similarly, changes in refractive index in the glass fiber decrease with the increasing Young's modulus and Poisson's ratio of the secondary coating but increase with the increasing Young's modulus and Poisson's ratio of the primary coating. Therefore, to minimize microbending loss induced simultaneously by axial strain and hydrostatic pressure in the glass fiber, the polymeric coatings should be suitably selected. An optimal design procedure is also indicated.

Journal Article↗

Effect of hydrostatic pressure of a mutant of Saccharomyces cerevisiae deleted in the trehalose-6-phosphate synthase gene.

Mutants Saccharomyces cerevisiae deleted on the trehalose-6-phosphate synthase gene (tps1) and their parental wild-type cells were submitted to hydrostatic pressure in the range of 0-200 MPa. Experimental evidence showed that viability for both strains decreased with increasing pressure and that tps1 mutants, unable to accumulate trehalose, were more sensitive to hydrostatic pressure than the wild-type cells. Additionally, both tps1 and wild-type cells in the stationary phase, when there is an accumulation of endogenous trehalose, were more resistant to pressure than proliferating cells. Under these conditions, mutant cells were also more sensitive to pressure treatment than the wild type. The present work also showed that mild pressure pretreatment did not induce hydrostatic pressure resistance (barotolerance) in yeast cells.

Gene Deletion↗

Differential regulation of stress proteins by high hydrostatic pressure, heat shock, and unbalanced calcium homeostasis in chondrocytic cells.

High hydrostatic pressure (HP) has recently been shown to increase cellular heat shock protein 70 (Hsp70) level in a specific way that does not involve transcriptional activation of the gene, but rather the stabilisation of the mRNA for Hsp70. In this study, we investigated whether there are other observable changes caused by HP stress, and compared them with those induced by certain other forms of stressors. A chondrocytic cell line T/C28a4 was exposed to 30 MPa continuous HP, heat shock at 43 degrees C, and increased cytosolic calcium concentration by the addition of sarco-endoplasmic reticulum Ca(2+) ATPase inhibitor thapsigargin (25 nM) or calcium ionophore A23187 (1 microM) in the cultures. The protein synthesis was studied by in vitro metabolic labelling followed by one- and two-dimensional polyacrylamide gel electrophoresis, and mass spectrometry was utilized to confirm the identity of the protein spots on two-dimensional gels. Continuous 30 MPa HP increased remarkably the relative labelling of Hsp70. Labelling of Hsp90 was also increased by 15-20%, although no clear change was evident at the protein level in Western blots. Elevated intracellular Ca(2+) concentration induced by thapsigargin and calcium ionophore A23187 increased mainly the synthesis of glucose-regulated protein 78 (Grp78/BiP), whereas Hsp70 and Hsp90 were decreased by the treatment. Heat shock was the strongest inducer of Hsp70 and Hsp90. This study further confirmed the induction of Hsp70 in chondrocytic cells exposed to high HP, but it also showed that calcium-mediated responses are unlikely to cause the stress response observed in the hydrostatically pressurized cells.

Amino Acid Sequence↗

TEMPERATURE-HYDROSTATIC PRESSURE STUDIES ON PARTIALLY PURIFIED INORGANIC PYROPHOSPHATASE ACTIVITY.

Morita, Richard Y. (Oregon State University, Corvallis), and Paul F. Mathemeier. Temperature-hydrostatic pressure studies on partially purified inorganic pyrophosphatase activity. J. Bacteriol. 88:1667-1671. 1964.-Partially purified inorganic pyrophosphatase from Bacillus stearothermophilus demonstrated increased activity at 90 C with hydrostatic pressures of 100 to 1,100 atm. At 90 C, optimal activity was at 700 atm. No enzyme activity could be demonstrated at 100 C at 1 atm; however, at 105 C moderate hydrostatic pressures favored the enzyme reaction, resulting in increased activity. The increased enzyme activity is explained on the basis of pressure counteracting the molecular volume increase which results from elevated temperature. Thermal denaturation studies at 90 C show inorganic pyrophosphatase to be more stable in the presence of cofactor than substrate, and indicate an enyzme-cofactor intermediate in the hydrolysis.

Animals↗

Saccharides that protect yeast against hydrostatic pressure stress correlated to the mean number of equatorial OH groups.

Several saccharides were found to be significantly effective in providing protection against hydrostatic pressure and high temperature damage in the yeast Saccharomyces cerevisiae. The extent of barotolerance and thermotolerance with seven different sugars showed a linear relationship to their mean number of equatorial OH groups. The same linear relationship is seen when sugars protect protein molecules against elevated temperatures in vitro. Some sugars were more effective in providing protection against hydrostatic pressure nearly a hundred times than high temperature. Pre-heat shock treatment on yeast cells induce various stress tolerances. In this report, pre-heat shocked cells showed potent protection against elevated temperature, but these cells showed faint protection against elevated pressure. These results suggest that sugars may protect cells against hydrostatic pressure and high temperature in a similar manner, probably by stabilizing the macromolecule(s), and such type of protection may be suited for pressure stress.

Carbohydrates↗