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In vivo hydraulic conductivity of muscle: effects of hydrostatic pressure.

We and others have shown that the loss of fluid and macromolecules from the peritoneal cavity is directly dependent on intraperitoneal hydrostatic pressure (Pip). Measurements of the interstitial pressure gradient in the abdominal wall demonstrated minimal change when Pip was increased from 0 to 8 mmHg. Because flow through tissue is governed by both interstitial pressure gradient and hydraulic conductivity (K), we hypothesized that K of these tissues varies with Pip. To test this hypothesis, we dialyzed rats with Krebs-Ringer solution at constant Pip of 0.7, 1.5, 2.2, 3, 4.4, 6, or 8 mmHg. Tracer amounts of 125I-labeled immunoglobulin G were added to the dialysis fluid as a marker of fluid movement into the abdominal wall. Tracer deposition was corrected for adsorption to the tissue surface and for local loss into lymphatics. The hydrostatic pressure gradient in the wall was measured using a micropipette and a servo-null system. The technique requires immobilization of the tissue by a porous Plexiglas plate, and therefore a portion of the tissue is supported. In agreement with previous results, fluid flux into the unrestrained abdominal wall was directly related to the overall hydrostatic pressure difference across the abdominal wall (Pip = 0), but the interstitial pressure gradient near the peritoneum increased only approximately 40% over the range of Pip = 1.5-8 mmHg (20-28 mmHg/cm). K of the abdominal wall varied from 0.90 +/- 0.1 x 10(-5) cm2.min-1.mmHg-1 at Pip = 1.5 mmHg to 4.7 +/- 0.43 x 10(-5) cm2.min-1.mmHg-1 on elevation of Pip to 8 mmHg. In contrast, for the same change in Pip, abdominal muscle supported on the skin side had a significantly lower range of fluid flux (0.89-1.7 x 10(-4) vs. 1.9-10.1 x 10(-4) ml.min-1.cm-2 in unsupported tissue). The differences between supported and unsupported tissues are likely explained in part by a reduced pressure gradient across the supported tissue. In conclusion, the in vivo hydraulic conductivity of the unsupported abdominal wall muscle in anesthetized rats varies with the superimposed hydrostatic pressure within the peritoneal cavity.

Abdominal Muscles↗

Hydrostatic pressure of the suprachoroidal space.

The hydrostatic pressure of the suprachoroidal space was measured in 18 cynomolgus monkey eyes by one of two methods: (1) direct cannulation, or (2) silicone sponge implantation. The intraocular pressure (IOP) and suprachoroidal pressure were monitored simultaneously with the IOP being held at various levels between 5 and 60 mm Hg. In eyes with direct cannulation, at an IOP of 15 mm Hg, the pressure in the anterior suprachoroidal (supraciliary) space was 0.8 +/- 0.2 mm Hg (n = 6, mean +/- SE) below the IOP, but the posterior suprachoroidal pressure was 3.7 +/- 0.4 mm Hg (n = 8) below the IOP. The suprachoroidal pressure in eyes with silicone sponge implant was 4.7 +/- 0.6 (n = 7) mm Hg below the IOP. A change in IOP produced a corresponding change in the supraciliary space pressure. However, the pressure difference between the anterior chamber and the posterior suprachoroidal space increased at higher IOP. This pressure differential is the driving force for uveoscleral outflow.

Animals↗

ATP is released from rabbit urinary bladder epithelial cells by hydrostatic pressure changes--a possible sensory mechanism?

1. The responses of rabbit urinary bladder to hydrostatic pressure changes and to electrical stimulation have been investigated using both the Ussing chamber and a superfusion apparatus. These experiments enabled us to monitor changes in both ionic transport across the tissue and cellular ATP release from it. 2. The urinary bladder of the rabbit maintains an electrical potential difference across its wall as a result largely of active sodium transport from the urinary (mucosal) to the serosal surface. 3. Small hydrostatic pressure differences produced by removal of bathing fluid from one side of the tissue caused reproducible changes in both potential difference and short-circuit current. The magnitude of these changes increases as the volume of fluid removed increases. 3. Amiloride on the mucosal (urinary), but not the serosal, surface of the membrane reduces the transepithelial potential difference and short-circuit current with an IC50 of 300 nM. Amiloride reduces the size of, but does not abolish, transepithelial potential changes caused by alterations in hydrostatic pressure. 4. Field electrical stimulation of strips of bladder tissue produces a reproducible release of ATP. Such release was demonstrated to occur largely from urothelial cells and is apparently non-vesicular as it increases in the absence of calcium and is not abolished by tetrodotoxin. 5. It is proposed that ATP is released from the urothelium as a sensory mediator for the degree of distension of the rabbit urinary bladder and other sensory modalities.

Adenosine Triphosphate↗

Inactivation of Mucor plumbeus by the combined actions of chitinase and high hydrostatic pressure.

Sporangiospores were treated with high hydrostatic pressure and/or fungal chitinase in order to study the inhibition of germination and growth of the food spoiling mold Mucor plumbeus. Total fungal inhibition was obtained either at 4.0 kbar or by 10 U/ml of chitinase from Penicillium janthinellum. A pretreatment with 1 U/ml of the same chitinase reduced the pressure necessary to obtain complete spore inhibition to 3 kbar.

Chitinases↗

The influence of tissue hydrostatic pressure and protein concentration on fluid and protein uptake by diaphragmatic initial lymphatics; effect of calcium dobesilate.

Net uptake of fluid and protein by rat diaphragmatic initial lymphatics was observed, at various tissue hydrostatic pressures (-5 to 25 cm water)--using the peritoneal cavity as a huge tissue space. Between -5 and +5 cms, the amount of fluid uptake was much increased as the protein concentration in the tissue fluid rose. This is as predicted by the colloidal osmotic pressure hypothesis of initial lymphatic filling; it is completely contrary to the hydrostatic pressure gradient hypothesis. However at 15 and 25 cm, the protein concentration had no effect on the fluid uptake. The initial lymphatics were acting as simple conduits, rather than as force-pumps. Fluid flowed down a hydrostatic pressure gradient, from the tissues to the collecting lymphatics. Fluid uptake was not directly proportional to the hydrostatic pressure gradient; it increased disproportionately at higher pressures--probably because of the dilatation of the initial lymphatics or their open junctions. Calcium dobesilate had no effect on fluid or protein uptake at the lower pressures, but significantly increased both of these at 25 cm of water. Probably it caused the collecting lymphatics to pump more, removing excess lymph which otherwise accumulated in them. This reduced the intralymphatic pressures in the initial lymphatics so that more fluid entered them.

Animals↗

Effects of cyclic hydrostatic pressure on proteoglycan synthesis in cultured chondrocytes and articular cartilage explants.

Primary chondrocyte cell cultures and explants of bovine articular cartilage were subjected to cyclic hydrostatic pressure in a novel computer-controlled pressure chamber designed for this purpose. The cultures were labeled with 5 microCi/ml 35SO4 and simultaneously pressurized with 5 MPa load for 1.5 or 20 h with pressure cycles of 0.0167, 0.05, 0.25, and 0.5 Hz. The chondrocyte cell cultures were also subjected to 0.0082 and 0.0034 Hz cycles. Sulfate incorporation was significantly inhibited in cell cultures subjected to the 0.5, 0.25, or 0.05 Hz cyclic loads for 1.5 h, but stimulated in explant cultures with a 0.5 Hz cyclic 1.5-h load. Chondrocyte cultures subjected to longer (20 h) loading showed a stimulation of sulfate incorporation with 0.5 and 0.25 Hz cycles, but an inhibition with 0.0167 Hz. The results indicate that cyclic hydrostatic pressures of presumably physiological magnitude have significant influences on proteoglycan synthesis in articular cartilage chondrocytes. Comparison of the cell and explant cultures under identical pressure conditions suggested that chondrocyte interactions with extracellular matrix are involved in this regulation by cyclic hydrostatic pressure. The responses of the chondrocytes to pressurization also varied according to the total length of the treatment, a finding compatible with the idea of multiple metabolic steps in chondrocytes, both pre- and post-translational, controlled by the ambient hydrostatic pressure.

Animals↗

Genomic expression pattern in Saccharomyces cerevisiae cells in response to high hydrostatic pressure.

Gene expression patterns in response to hydrostatic pressure were determined by whole genome microarray hybridization. Functional classification of the 274 genes affected by pressure treatment of 200 MPa for 30 min revealed a stress response expression profile. The majority of the >2-fold upregulated genes were involved in stress defense and carbohydrate metabolism while most of the repressed ones were in cell cycle progression and protein synthesis categories. Furthermore, uncharacterized genes were among the 10 highest expressed sequences and represented 45% of the total upregulated genes. The results of this study revealed a hydrostatic pressure-specific stress response pattern and suggested interesting information about the mechanisms involved in adaptation of cells to a high-pressure environment.

Energy Metabolism↗

ERK 1/2 activation in enhanced osteogenesis of human mesenchymal stem cells in poly(lactic-glycolic acid) by cyclic hydrostatic pressure.

The aim of this study was to identify the signal transduction pathways and mechano-transducers that play critical roles in the processes induced by changes in cyclic hydrostatic pressure and fluid shear in 3-dimensional (3D) culture systems. Mesenchymal stem cells were loaded into a polymeric scaffold and divided into three groups according to the stress treatment: static, fluid shear, and hydrostatic pressure with fluid shear. Cells were exposed daily to a hydrostatic pressure of 0.2 MPa for 1 min followed by 14 min rest with fluid flow at 30 rpm. Protein extracts were analyzed by Western blot for extracellular signal-regulated kinase 1/2 (ERK1/2). The complexes were cultured under the mechanical stimuli for 21 days with or without phospho-ERK1/2 inhibitor (U0126) and evaluated by RT-PCR, calcium contents, and immunohistochemistry. Under conditions of mechanical stimulation, the activation of ERK1/2 was sustained or increased with time. U0126 suppressed mechanical stimuli-induced expression of osteocalcin. In addition, calcium contents and the degrees of osteocalcin and osteopontin staining were decreased by this inhibitor. These results demonstrate that mechanical stimuli, particularly hydrostatic pressure with fluid shear, enhance osteogenesis in 3D culture systems via ERK1/2 activation.

Cell Culture Techniques↗

The influence of hydrostatic pressure on the distribution of histone mRNA in HeLa cells.

Hydrostatic pressure and HeLa S3 cells were used (as a model system) to investigate the relationship of the cytoskeleton and histone gene expression. Exposure of HeLa S3 cells to hydrostatic pressure of 1000 - 10,000 psi (6.89 x 10(3) - 6.89 x 10(4) kPa) disrupts the cytoskeleton and reduces H1 and core histone mRNA and actin mRNA levels as determined by hybridization to specific DNA probes. Soluble and insoluble cell fractions were isolated from HeLa cells after lysis in Triton X-100 buffered with PIPES and being subjected to low-speed centrifugation. The insoluble fraction was designated the cytoskeletal fraction. At atmospheric pressure, 76% of H4 histone mRNA is associated with the cytoskeletal fraction and 24% of the H4 histone mRNA is in the soluble fraction. At 6000 and 10,000 psi for a duration of 10 min, H4 mRNA levels in the cytoskeletal fraction were reduced to 52 and 41%, respectively. The reduction of mRNA in the cytoskeletal fraction is accompanied by a corresponding increase of mRNA in the soluble cell fraction. The other core (H2A, H2B, and H3) and H1 histone mRNA transcripts exhibited similar sensitivity to pressure treatment. The effects of pressure on histone gene regulation may be mediated through alteration of mRNA-cytoskeleton association.

Actin Cytoskeleton↗

Evolution of intraperitoneal hydrostatic pressure following peritoneal catheter implantation.

Intraperitoneal hydrostatic pressure (IPP) was measured in 10 patients for 15 days (until day 15) following peritoneal catheter implantation. Tenckhoff catheters were surgically implanted using a transrectal approach. The patients began peritoneal dialysis (PD) immediately, initially with small intraperitoneal fluid volumes (500 mL), gradually increasing to reach 2000 mL by day 7. In one patient PD was delayed until day 7 because of an initial leakage of dialysate. Inspiratory and expiratory IPP were measured daily, with the zero point fixed at the midaxillary line with the patient in the supine position. Despite the small intraperitoneal fluid volumes used initially (500 mL), IPP was much higher in the first 3 days following implantation than in the next 12 days (17 +/- 3 cm of water vs 11 +/- 1 cm of water). IPP remained high until day 3, then gradually decreased in a linear fashion until day 12, despite the increased intraperitoneal volumes used, and then remained constant. In conclusion, IPP was found to be significantly higher for 3 days following intraperitoneal catheter implantation, whichever intraperitoneal volume was used. This may be due to tense abdominal muscles from pain caused by the surgery or because of reduced intraperitoneal compliance at the beginning of PD.

Adult↗

Influence of hydrostatic pressure on the effects of the heavy metal cations of manganese, copper, cobalt, and nickel on the growth of three deep-sea bacterial isolates.

Increases hydrostatic pressure varied the 72-h growth yield of three bacterial isolates from the deep sea in the presence of heavy metal cations of Mn, Cu, Co, and Ni, depending on the bacterial isolate, the metal cation and its concentration, and the level of hydrostatic pressure. Above atmospheric, hydrostatic pressure was found to have one of the following four effects on the response of culture growth to a heavy metal cation. (i) It could be without effect; (ii) it could enhance inhibition by a metal cation; (iii) it could increase the 72-h growth yield by a metal cation; or (iv) it could protect against a growth inhibitory effect noted at a lower pressure. Possible reasons for these varied responses are discussed.

Bacteria↗

[Effects of high hydrostatic pressure on energy metabolism of Lactobacillus plantarum].

Effects of high hydrostatic pressure on energy metabolism were investigated with Lactobacillus plantarum ATCC8014 as the test microorganism in this work. An INT colorimetric method of oxidation-reduction was established to measure INT metabolic activity of deoxidization of L. plantarum ATCC8014 cells. The utilization of glucose and INT metabolic activity of deoxidization of the cells after HPP treatment were determined using colorimetric methods. The experimental results showed that survival counts of ATCC8014 cells on MRS agar medium and INT metabolic activity of deoxidization decreased significantly, and little changes of utilization of glucose took place with increasing pressure ranging from 150 to 250 MPa for 15 min. Utilization of glucose also reduced evidently at high pressure ( > 300 MPa) for 15 min. Whereas survival cell counts on MRS agar medium were below the detection limit and INT metabolic activity of deoxidization of ATCC8014 was 0% after a 15-min pressure holding time at 400MPa, utilization of glucose of the cells retained 56.1% compared with that of untreated cells. In summary, it can be concluded that enzymes absorbing and transporting glucose in cellular membrane appear to have a high resistance to pressure, enzymes and biological regulating systems involved in glycolysis are more resistant to pressure than those in TCA (tricarboxylic acid cycle) system, TCA of ATCC8014 is more sensitive to pressure than glycolysis, and the decrease of INT metabolic activity of deoxidization is highly related to cell reduction during HHP, which provide some theoretical evidences for mechanisms of HHP sterilization. Inhibition of TCA metabolism is a very important cause of ATCC8014 inactivation by HHP. High hydrostatic pressure can be used as an effective tool to explore pathways of biological metabolism.

Energy Metabolism↗

Prostaglandin E2 induced changes in renal blood flow, renal interstitial hydrostatic pressure and sodium excretion in the rat.

Prostaglandin E2, when infused into the renal artery of the dog, is a vasodilator and increases both renal interstitial hydrostatic pressure and sodium excretion. Similar studies in the rat, however, have been inconclusive. The present study examined the effect of prostaglandin E2 infusion into the renal interstitium, by means of a chronically implanted matrix, on renal blood flow, renal interstitial hydrostatic pressure and sodium excretion in the rat. Prostaglandin E2 was continuously infused directly into the kidney interstitium to mimic endogenous prostaglandin E2 production by renal cells. The maximum change in each of these parameters occurred when 10(-5) M PGE2 was infused. Renal blood flow increased from 4.70 +/- 0.91 to 5.45 +/- 0.35 ml/min (p less than 0.05) while renal interstitial hydrostatic pressure decreased from 3.9 +/- 0.4 to 2.6 +/- 0.5 mmHg (p less than 0.05) and fractional excretion of sodium decreased from 1.02 +/- 0.20 to 0.61 +/- 0.12% (p less than 0.05). Thus, the present study demonstrates that renal interstitial infusion of prostaglandin E2 increases total renal blood flow but decreases both renal interstitial hydrostatic pressure and urinary sodium excretion in the rat.

Animals↗

Fate of Escherichia coli strains inoculated in model cheese elaborated with or without starter and treated by high hydrostatic pressure.

The aim of this research was to study high hydrostatic pressure inactivation of two strains of Escherichia coli (E. coli O59:H21 [CECT 405] and E. coli O157:H7 [CECT 5947]) inoculated in washed-curd model cheese elaborated with and without starter and the ability of these strains for survival, recovery, and growth. Samples were treated at 300, 400, and 500 MPa for 10 min at 20 degrees C and analyzed after the treatment and after 1, 7, and 15 days of storage at 8 degrees C to study the behavior of Escherichia populations. Cheeses elaborated with starter showed the maximum lethality at 400 and 500 MPa, and no significant differences in the baroresistant behavior of either strains were detected, except for E. coli O157:H7 at 400 MPa in cell counts obtained with thin agar layer method medium, where the decrease value was significantly lower. In cheese elaborated without starter, the highest decrease value was observed at 500 MPa, except for E. coli O59:H21 in cell counts obtained with selective culture medium, where the highest decrease value was also found at 400 MPa. The ability to repair and grow was not observed in model cheese elaborated with starter, as cell counts of treated samples decreased after 15 days of storage at 8 degrees C. By contrast, in cheese elaborated without starter, all pressurized samples showed the trend to repair and grow during the storage period in both strains. These results suggest that the presence of starter and low pH values are the main factors that control the ability of Escherichia strains inoculated in this type of cheese and treated by high hydrostatic pressure to recover and grow.

Cheese↗

Effects of hydrostatic pressure on matrix synthesis in different regions of the intervertebral disk.

The intervertebral disk is routinely subjected to compressive loads that alter with posture and muscle activity and can produce pressures > 2 MPa in human lumbar disks in vivo (A. Nachemson and G. Elfstrom. Scand. J. Rehabil. Med. 2, Suppl. 1:1-40, 1979; A. Nachemson and J. M. Morris. J. Bone Jt. Surg. Am. Vol. 46A: 1077-1092, 1964). We measured the effect of load on hydrostatic pressures in bovine caudal disks. With increase in applied load, pressure increased linearly in the nucleus and inner annulus. The resting pressure measured after slaughter (0.19 +/- 0.05 MPa) and the pressure at failure (34 MPa, estimated from the vertebrae/disk segment failure load of 7,430 +/- 590 N) define the limits that can occur in vivo. Because hydrostatic pressure influences matrix synthesis in articular cartilage, we have examined the effects of pressures in the range 1-10 MPa applied for 20 s or 2 h on proteoglycan synthesis in bovine caudal and human lumbar intervertebral disks in vitro. In the nucleus pulposus and inner annulus of bovine disks, application of hydrostatic pressure in the range of 1-7.5 MPa for only 20 s stimulated matrix synthesis over the following 2 h at atmospheric pressure. The maximum stimulation in the bovine disks was seen in the inner annulus after application of 2.5 MPa, where proteoglycan synthesis rates doubled. Exposure to 2.5 MPa also stimulated synthesis in the nucleus pulposus of human disks taken at surgery, whereas 7.5 MPa inhibited synthesis in five out of six specimens. With 2-h continuous exposure to the same levels of pressure, no stimulation was seen in the nucleus of bovine disks, and significant stimulation was only observed at 5.0 MPa in the inner annulus. Exposure to 10 MPa for either 20 s or 2 h inhibited proteoglycan synthesis in these regions of the disks. In contrast, in the outer annulus, where loading does not lead to a rise in hydrostatic pressure in vivo, there was no significant response to hydrostatic pressure over the range of 1-10 MPa in bovine or human disks.

Animals↗

A new experimental model for studying the response of periodontal ligament cells to hydrostatic pressure.

An apparatus was developed to apply positive or negative hydrostatic pressure dynamically to periodontal ligament (PDL) cells in vitro. The objective of this investigation was to construct this apparatus and to determine its effects on PDL cells. Human PDL cells were collected from freshly extracted premolars. At the sixth passage, the cells were mechanically stimulated by this apparatus at different magnitudes of continuous positive or negative hydrostatic pressures (PHP or NHP, respectively). The application of PHP between 0.3 and 30 gm/cm2 significantly enhanced prostaglandin E (PGE) production and intracellular cyclic AMP (cAMP) of the cells. In contrast, perturbation by NHP significantly decreased PGE production and intracellular level of cAMP. Proliferation rate increased significantly at 24 and 48 hours due to stimulation of these cells with -30 gm/cm2 of NHP. Challenging these cells with +30 gm/cm2 of PHP significantly decreased the proliferation rate of these cells at 24 and 48 hours. Stimulation by PHP between +30 to +600 gm/cm2 increased cell length and width and appeared to increase surface area attachment to the bottom of the culture dishes. In contrast, NHP (between -30 and -600 gm/cm2) decreased these dimensions and appeared to reduce the surface area of attachment. These results indicate that this type of mechanical perturbation of PDL cells produces physiologic responses and is not detrimental to their vitality.

Atmosphere Exposure Chambers↗

Measurement of hydrostatic pressures during simulated post cementation.

Tooth sensitivity and fracture after cementation of posts for endodontically treated teeth have been a problem. This investigation developed an in vitro method of measuring intraradicular hydrostatic pressures created during simulated post cementation. The testing apparatus consisted of a pressure transducer and brush recorder connected to precision milled post spaces in a Plexiglas block. Cast post and cores were fabricated and cemented with three different luting agents: resinous cement, glass ionomer cement, and zinc phosphate cement. Mean hydrostatic pressures (psi) recorded during post cementation were zinc phosphate cement, 22.67; resinous cement, 19.77; and glass ionomer cement, 17.66. Zinc phosphate cement created substantially greater hydrostatic pressures than either the resinous or glass ionomer cements. This in vitro system was capable of discriminating intraradicular hydrostatic pressures among different classes of luting agents.

Analysis of Variance↗

August Krogh Lecture. The renal concentrating mechanism in insects and mammals: a new hypothesis involving hydrostatic pressures.

Water moves from compartments of higher to compartments of lower water potential. Osmotically active solutes and negative hydrostatic pressure both lower water potential by stretching the hydrogen bonds between water molecules (Hammel-Scholander hypothesis). In trees the negative hydrostatic pressure in the sap is balanced by the osmotic pressure of the leaves. In response to differences in water potential, water flows across biological membranes through water-filled pores. Protein molecules, aquaporins, forming hourglass-shaped pores have been identified, cloned, and located in plasma membranes in mammalian as well as other tissues. Water molecules flow single file through aquaporins. Insects concentrate the urine in the rectum. Mammals concentrate the urine in the collecting ducts in the inner medulla. In both, a compartment with a high osmotic concentration is created through ion transport. Both have a muscular coat surrounding the tissue, which shows peristaltic contractions. In insects it is the muscular layer around the rectum; in mammals it is the renal pelvic wall that surrounds the papilla. Mechanisms are proposed whereby these peristaltic contractions, through the creation of positive and negative hydrostatic pressures in the tissues, can lead to hyperosmotic excreta.

Animals↗