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At least 19 recordsLinked to original sources
PubMed1990

Effects of retrograde flow on measured blood volume, Disse space, intracellular water space and drug extraction in the perfused rat liver: characterization by the multiple indicator dilution technique.

Previous rat liver perfusion studies had shown a preferential derecruitment of downstream (phenacetin O-de-ethylation) in relation to upstream (acetaminophen sulfation) hepatocyte activities with reductions in prograde (normal) flow, with substrates entering from the portal vein and exiting the hepatic vein. The reduction in metabolic activities was found associated with a reduction in accessible intracellular water space (Pang et al., J. Pharmacokinet. Biopharm. 16: 595, 1988). In the present studies, we investigated the changes in metabolic activities of the liver and its accessible spaces during retrograde flow (substrates entering from the hepatic vein and exiting the portal vein), an approach used to probe zonal heterogeneity in the liver. The multiple indicator dilution technique was used to gain information concerning the accessible sinusoidal blood volume, Disse space, and the intracellular water space; once-through perfusion with tracer [3H]acetaminophen and [14C]phenacetin was performed simultaneously within the same liver, to estimate metabolic activities at the periportal and perihepatic venous regions of the liver, respectively, at the designated flows. The estimated sinusoidal blood volume, total albumin and sucrose distribution spaces, the Disse space and the accessible albumin and sucrose Disse spaces showed weak but positive correlations with retrograde perfusate flow rate. The values were substantially greater than those for rat livers perfused with normal (prograde) flows, suggesting an increase in the vascular space and accessible Disse space of the liver during retrograde flow. The total water and accessible intracellular water spaces also showed weak but positive correlations with retrograde perfusate flow, and the average intracellular water space (which is also accessed by the substrates) at both retrograde flows of 8 and 12 ml/min was 0.52 +/- 0.08 and 0.55 +/- 0.08 ml/g liver, respectively, values that were not statistically different. When the intracellular water space was plotted vs. water flow rate (ml water/g liver), a weakly positive correlation was again found, suggesting that over the flow range, the accessible intracellular water space decreased in a very gradual and linear fashion with a reduction in water flow rate. When the estimated intracellular water space for retrograde flow was compared with values obtained for the same flow rate in a prograde fashion, slightly lower values were obtained. After consideration of the effect of the distended vascular and interstitial spaces on the weights of the liver, the values obtained for retrograde and prograde flow rates were closely similar. In both cases, it appeared that with reduction in flow, the accessible intracellular water spaces were reduced, regardless of the flow direction.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetaminophen↗
PubMed1988

Effects of perfusate flow rate on measured blood volume, disse space, intracellular water space, and drug extraction in the perfused rat liver preparation: characterization by the multiple indicator dilution technique.

The effect of hepatic blood flow on the elimination of several highly cleared substrates was studied in the once-through perfused rat liver preparation. A constant and low input concentration of ethanol (2.0 mM), [14C]-phenacetin and [3H]-acetaminophen (0.36 and 0.14 microM, respectively), or meperidine (8.1 microM) was delivered once-through the rat liver preparation in five flow periods (greater than 35 min each); control flow periods at 12 ml/min were interrupted by flow changes to 8 or 16 ml/min. The steady-state hepatic availabilities (F or outflow survivals) at 12 ml/min were ethanol, 0.075 +/- 0.038; [14C]-phenacetin, 0.15 +/- 0.059; [3H]-acetaminophen, 0.34 +/- 0.051; meperidine, 0.047 +/- 0.017. Flow-induced changes were different among the compounds: with reduced flow (8 ml/min), F was decreased for ethanol (0.061 +/- 0.032) and [3H]-acetaminophen (0.28 +/- 0.051), as expected, but was increased for [14C]-phenacetin (0.20 +/- 0.068) and meperidine (0.05 +/- 0.03); with an elevation of flow (to 16 ml/min), F was increased for all compounds, as expected of shorter sojourn times: ethanol, 0.13 +/- 0.065; [14C]-phenacetin, 0.22 +/- 0.062; [3H]-acetaminophen, 0.43 +/- 0.063; meperidine, 0.055 +/- 0.022. A marked increase in F for ethanol had occurred when flow changed from 12 to 16 ml/min due to nonlinear metabolism; the latter was confirmed by a reduction in the extraction ratios at increasing concentrations (1.8 to 11.4 mM); this condition was not present for the other compounds. In order to explain the observations, we used the multiple indicator dilution technique to investigate the flow-induced behaviors of tissue distribution spaces of vascular and intracellular references in the perfused rat liver preparation.

Acetaminophen↗
PubMed2006

Active mechanical stabilization of the viscoplastic intracellular space of Dictyostelia cells by microtubule-actin crosstalk.

The micro-viscoelasticity of the intracellular space of Dictyostelium discoideum cells is studied by evaluating the intracellular transport of magnetic force probes and their viscoelastic responses to force pulses of 20-700 pN. The role of the actin cortex, the microtubule (MT) aster and their crosstalk is explored by comparing the behaviour of wild-type cells, myosin II null mutants, latrunculin A and benomyl treated cells. The MT coupled beads perform irregular local and long range directed motions which are characterized by measuring their velocity distributions (P(v)). The correlated motion of the MT and the centrosome are evaluated by microfluorescence of GFP-labelled MTs. P(v) can be represented by log-normal distributions with long tails and it is determined by random sweeping motions (v approximately 0.5 microm/s) of the MTs (caused by tangential forces on the filament ends coupled to the actin cortex) and by intermittent bead transports parallel to the MTs (v(max) approximately 1.5 microm/s). The tails are due to spontaneous filament deflections (with speeds up to 10 microm/s) attributed to pre-stressing of the MT by local cortical tensions, generated by dynactin motors generating plus-end directed forces in the MTs. The viscoelastic responses are strongly non-linear and are mostly directed opposite or perpendicular to the force, showing that the cytoplasm behaves as an active viscoplastic body with time and force dependent drag coefficients. Nano-Newton loads exerted on the soft MT are balanced by traction forces arising at the MT ends coupled to the actin cortex and the centrosome, respectively. The mechanical coupling between the soft microtubules and the viscoelastic actin cortex provides cells with high mechanical stability despite the softness of the cytoplasm.

Actins↗
PubMed1990

Suppression of water shift into intracellular space by TA-3090 (calcium entry blocker) measured with NMR.

In order to clarify the effects of TA-3090 (calcium entry blocker) on the suppression of water after ischaemic events, 16 measurements of T2f were continuously performed on brain biopsy (for 2-60 min) obtained from treated and control Wistar rats. The time constant (k) and NMR parameters (T2f,(O) delta T2f, T2fmax(T2f(O) + delta T2f) were obtained from 16 values of T2f. The values of k in Wistar rats treated with intravenously administrated TA-3090 (0.5 mg/kg) were significantly prolonged as compared to that of control. There were no significant differences of maximum prolongation of T2f(T2fmax(T2f(O) + delta T2f) among three groups. Since the prolongation of T2f after biopsy reflects the water shift from extra to intracellular space, the increments of time constant indicates that TA-3090 suppresses the water shift into intracellular space. Our present results suggest that TA-3090 prevents some processes involved in irreversible cell damage and suppresses the cytotoxic brain oedema in the incomplete ischaemic area where non-competitive calcium channel is inactive.

Animals↗
PubMed1997

Phosphonates as 31P-NMR markers of extra- and intracellular space and pH in perfused rat liver.

We evaluated phosphonates (Po) as markers of the extra- and intracellular space in perfused rat liver. (i) In- and outwash behaviour of phenylphosphonate (PhePo), 3-amino-propylphosphonate (NProPo) and methyl phosphonate (MePo) was compared with that of creatine phosphate (CrP), a marker of the extracellular space, and of dimethyl methylphosphonate (MePoMe2), a marker of the total water-accessible space. In- and outwash of CrP was accurately predicted by the time constant (approximately 12 s) for the in- and outwash of inulin, a standard marker of the extracellular space. MePoMe2 rapidly distributed over the total liver volume (about three times the CrP accessible space). PhePo, NProPo and MePo washed rapidly into the extracellular space with CrP, and then steadily spilled over into the MePoMe2-accessible space. Upon outwash, Po signals rapidly declined in phase with that of CrP. Residual Po (PhePo >> NProPo approximately equal to MePo) reflected the amount internalized during prolonged (60 min) inwash. Proportional amounts of residual Po were found in extracts of livers harvested after outwash of perfusate and extracellular markers. Consistent with exclusion from the cells, CrP went undetected in these extracts. (ii) The resonance frequency of residual PhePo after outwash of the extracellular fraction corresponded with the pH reported by cytosolic P1 and responded to transient changes of the intracellular pH, induced by perfusion with and withdrawal of 20 mM NH4Cl. (iii) MePoMe2 homogeneously distributed over perfusate, parenchyma and bile, consistent with unrestricted permeability. Other Po were transported transcellularly and excreted in bile. CrP was virtually excluded from the bile, attesting to a minimal role for 'bulk-phase pinocytotic' transcellular transport, or for 'paracellular' leakage. In summary, charged Po can be used as extracellular markers in liver, provided experimental conditions are adjusted to minimize their internalization. Some Po (e.g. PhePo) can reach intracellular concentrations which suffice for the compound to act as a reporter molecule of the cytosolic pH.

Animals↗
PubMed1990

Effect of glutamate and its antagonist on shift of water from extra- to intracellular space after cerebral ischaemia.

The effects of glutamate and the excitatory amino acid antagonist, MK-801, were investigated on the time course of the shift of water from extracellular to intracellular space (progression or cytotoxic oedema) after total brain ischaemia in rats. Administration of sodium glutamate intravenously before ischaemia accelerated the shift of water dose-dependently. On the contrary, preischaemic administration of MK-801, an NMDA antagonist, delayed the progression of cytotoxic oedema due to brain ischaemia. We consider that glutamate and NMDA antagonists may have important roles in the development and prevention of cytotoxic oedema in the ischaemic state.

Animals↗
PubMed2002

Evidence that both fast and slow water ADC components arise from intracellular space.

Evaluation of water diffusion in the brain has revealed both fast- and slow-diffusing water populations. It has been suggested that these populations represent extra- and intracellular water, respectively. We have identified and characterized both populations in the intracellular space of the Xenopus oocyte. We have also determined their T(1) and T(2) relaxation properties. The fast and slow intracellular populations have diffusion coefficients of 1.06 +/- 0.05 microm(2)/ms and 0.16 +/- 0.02 microm(2)/ms, respectively, with the fast fraction representing 89% +/- 1% of the total water signal. These values are quite similar to those for total water in brain and are observed in the absence of signal from the perfusate (extracellular) water population. Volumetric swelling (16% +/- 4%) of the oocyte in hypoosmotic media increased the diffusion coefficients of both intracellular populations (fast = 1.27 +/- 0.03 microm(2)/ms, slow = 0.22 +/- 0.02 microm(2)/ms), but did not change their relative signal fractions. This phenomenon runs counter to the effects observed in brain injury, following which the apparent diffusion coefficient (ADC) decreases 30-50%. The results presented herein suggest that this ADC decrease in brain occurs despite cell swelling, which by itself would be expected to induce an increase in intracellular diffusion coefficients.

Animals↗
PubMed1977

Penetration of Mannitol into the Intracellular Space of Chlorella sorokiniana.

It was found that in the alga, Chlorella sorokiniana, mannitol penetrated the plasmalemma (normally thought to be impermeable to mannitol) into the intracellular space. The rate of penetration is exponential and relatively slow, having a half-time of 6 to 12 minutes and requiring over 60 minutes for complete penetration. This penetration was demonstrated both by the Millipore filtration of cells incubated with (14)C-mannitol and by centrifugation of the cells through a silicon oil layer after incubation with tritiated water and (14)C-mannitol. Further, mannitol caused an inhibition of both autotrophic (on CO(2)) and heterotrophic (on glucose) growth. A low rate of mannitol metabolism was demonstrated, although this rate could not support heterotrophic growth.

Journal Article↗
PubMed1995

On the use of 133Cs as an NMR active probe of intracellular space in vivo.

Data are presented from 133Cs NMR studies on both excised and in situ tissues from rats fed a regular diet and administered i.p. CsCI in aqueous solution for 6 to 14 days. Cesium is an NMR-active potassium analog which accumulates in the intracellular spaces of tissues [Davies et al., Biochemistry 27, 3547 (1988); Shehan, B.P. et al., Magn. Reson. Med. 30,573 (1993)]. Chemical shifts, relaxation properties, sensitivity and detectability of cesium in tissues were investigated. Consistent with previous reports, two resonances (representing intra- and extracellular cesium) were detected in blood. Only one resonance was detected in brain, kidney, and muscle tissue. Efforts to resolve intra- and extracellular components by T1 and T2 relaxation discrimination were not successful. Following i.p. administration, cesium accumulates intracellularly with a brain-to-cerebrospinal fluid concentration (mumol/g) ratio of 9:1 and a thigh muscle-to-plasma concentration ratio of 40:1. Considering the small extracellular volume in these tissues (ca 18% and 10%, respectively), the net content differences between intra- and extracellular cesium are approximately 40-fold in brain and 360-fold in muscle. The concentration ratio of cesium in brain to cesium in cerebrosinal fluid decreased to 3:1 1 h after death, indicating a relatively slow rate of leakage of cesium from the intra- to extracellular space in the face of bioenergetic failure. These data suggest that the cesium signal is dominated by the intracellularly located cesium and, thus, cesium may be useful as a probe of the intracellular environment despite an inability to resolve and directly observe distinct resonances from intra- and extracellular spaces.

Animals↗
PubMed2000

1-aminocyclopropane-1-carboxylate (ACC) deaminase induced by ACC synthesized and accumulated in Penicillium citrinum intracellular spaces.

We have already described how 1-aminocyclopropane-1-carboxylic acid (ACC), which is a precursor of the plant hormone ethylene, is synthesized in Penicillium citrinum through the same reaction by the catalysis of ACC synthase [EC 4.4.1.14] as in higher plants. In addition, ACC deaminase [EC 4.1.99.4], which degrades ACC to 2-oxobutyrate and ammonia, was also purified from this strain. To study control of induction of ACC deaminase in this organism, we have isolated and analyzed the cDNA of P. citrinum ACC deaminase and studied the expression of ACC deaminase mRNA in P. citrinum cells. By the analysis of peptides from the digests of the purified and modified ACC deaminase with lysylendopeptidase, 70 % of its amino acid sequences were obtained. These amino acid sequences were used to identify a cDNA, consisting of 1,233 bp with an open reading frame of 1,080 bp encoding ACC deaminase with 360 amino acids. The deduced amino acids from the cDNA are identical by 52% and 45% to those of enzymes of Pseudomonas sp. ACP and Hansenula saturnus. Through Northern blot analysis, we found that the mRNA of ACC deaminase was expressed in P. citrinum cells grown in a medium containing 0.05% L-methionine. These findings suggest that ACC synthesized by ACC synthase and accumulated in P. citrinum intracellular spaces can induce the ACC deaminase that degrades the ACC.

Amino Acid Sequence↗
PubMed1990

Influence of pretreatment on interstitial and intracellular space of canine left-ventricular myocardium.

In this study, the interstitial space and myocytes were investigated qualitatively and morphometrically in samples from beating, fibrillating as well as from cardioplegically HTK-arrested hearts fixed by immersion or perfusion. The size of tissue clefts separating bundles of myocytes and that of the interstitial space within bundles of myocytes depend on the functional state and on the kind of fixation. Cellular preservation is significantly better in HTK-arrested hearts compared to beating or fibrillating hearts. Thus, for the structural evaluation of myocytes and interstitium, the pretreatment constitutes a highly significant factor.

Animals↗