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PubMed1977

Concentrations of antidiuretic hormone in plasma during human sodium restriction.

Negative sodium balance was produced in 10 human volunteers. Body weight, plasma sodium, osmolality, hematocrit, renin activity (PRA), and antidiuretic hormone (ADH) concentrations were determined before, during, and after sodium restriction. Body weight declined and PRA rose during the period of low sodium intake. Plasma sodium concentration and osmolality did not change. A statistically significant change in ADH was not observed. It is suggested that a decrease in ADH was prevented by a rising titer of renin and contraction of the extracellular space.

Body Weight↗

PubMed1977

Ultrastructure of the liver in a case of childhood cystinosis.

Ultrastructural findings in the liver in a case of childhood cystinosis are reported. Crystalline structures were found mainly in Kupffer cells. The presence of dark cells, with or without crystals, was the most striking feature observed. Such cells have already been noted within the kidney on one occasion when it was shown that the dark substance was L-cystine (Spear et al., 1971). In this case identical dark material was also found extracellularly. The data shows that free cystine can fill cell cytoplasm and extracellular spaces and the possibility that cystine overproduction may take place in the hyaloplasm should be considered. Extracellular location of cystine in the tubules might account for an increase in epithelial permeability and thus for the Fanconi syndrome.

Child↗

PubMed2000

Aminomethylphosphonate and 2-aminoethylphosphonate as (31)P-NMR pH markers for extracellular and cytosolic spaces in the isolated perfused rat liver.

Aminomethylphosphonate (NMePo) and 2-aminoethylphosphonate (NEthPo) were evaluated as alternative pH indicators in the isolated perfused rat liver using (31)P-nuclear magnetic resonance (NMR). NMePo did not distribute within cells and remained in the extracellular space. It exhibited pH titration with a low pK(a) value (5.35). This behaviour makes NMePo useful for extracellular volume or acidic pH determination. In contrast, NEthPo accumulated within cells without altering liver energetic steady state, evaluated from nucleosides triphosphates resonances, even for prolonged (100 min) experiments. Withdrawal of NEthPo from perfusate revealed a residual resonance corresponding to the internalized amount of this phosphonate. This fraction was almost stable vs time and allowed determination of spin-lattice relaxation time constant T(1) within the liver (2.2 +/- 0.3 s; n = 6). Comparison of the titration curves for NEthPo and inorganic phosphate revealed that the accuracy of pH determination within physiologic or acidic range in both cases was comparable. Finally, when extracellular pH was decreased, the NEthPo resonance frequency was found to undergo the same chemical shift variations as observed for cytosolic P(i) signal, which was in good agreement with a cytosolic accumulation of this phosphonate. Therefore, NEthPo could be considered as an interesting cytosolic pH probe suitable for (31)P-NMR measurements, especially when experimental conditions prevent reliable observation of cytosolic Pi resonance.

Aminoethylphosphonic Acid↗

PubMed1977

Ultastructural analysis on acetylcholinesterase localization in the cerebellar cortex of teleosts.

The histochemical localization of acetylcholinesterase (AChE) was studied by electron microscopy in the cerebellar cortex of the goldfish and the catfish. The patterns of enzyme distribution show noticeable differences in the two teleost species at the level of the corresponding cerebellar structures. Among the most distinctive features is the prevailing intracellular localization of enzyme activity in the goldfish and the prevailing extracellular localization in the catfish in the molecular layer and, to a lesser extent, the granular layer. Only quantitative differences in the ability to synthesize AChE can be recorded among the different cerebellar neurons in the two species, since all these neurons exhibit different amounts of enzyme activity linked to their cytoplasmic structures. Comparing the results obtained with those of previous histochemical, experimental and developmental researches, the hypothesis seems well founded that the embryonic pool of cerebellar neurons is made up of AChE-synthesizing nruroblasts which, during development, loss or maintain to a different the mechanisms for AChE synthesis. In addition the light and electron microscope histochemistry reveals at different levels of resolution that the final pattern of AChE distribution in the cerebellar cortex is the sum of different degress of AChE synthesis by cerebellar neurons and different degrees of enzyme release in extracellular spaces.

Acetylcholinesterase↗

PubMed1985

Extracellular distribution of radiolabel obscures specific binding of diethylstilbestrol in mouse skeletal muscle.

The extracellular distribution of 3H-diethylstilbestrol (3H-DES) in mouse skeletal muscle was assessed following intraperitoneal injection. Total muscle extracellular space was measured with 14C-inulin, and the vascular space with 125I-albumin. A significant difference in the distribution of native 3H-DES and its metabolites in muscle and blood was found. This could only be explained if these compounds distributed with the albumin space and not the inulin space.

Albumins↗

PubMed1989

Age-related changes in the internal branch of the rat superior laryngeal nerve.

This investigation was initiated to provide data on the ultrastructural basis for neurologic age-related changes in laryngeal sensory function. In the present study, an animal model (female Wistar rats: age range: young [Y], 3 to 5 months; old [O], 25 months; and very old [VO], 29 to 30 months) was used to examine systematically changes in the internal branch of the superior laryngeal nerve with age using electron microscopic morphometric techniques. Total fiber counts, fiber populations (size categories), and mean fiber size for myelinated and unmyelinated fibers did not change with age. Qualitative changes were consistent with segmental demyelination and axonal degeneration in the older animals. There was also a significant age-related increase in the volume fraction of adaxonal Schwann cell cytoplasm (Y, 0.019; O, 0.041; and VO, 0.042). Ultrastructural correlates of intracellular support and axonal transport showed a significant decrease in the numerical density of neurofilaments (Y, 0.126/micron2; O, 0.073/micron2; and VO, 0.078/micron2) in the older animals, but no change in the numerical density of microtubules. Energy metabolism correlates in the form of mitochondrial volume fraction did not change with age. There was a significant increase in the volume fraction of the intrafascicular extracellular space (Y, 0.224; O, 0.271; and VO, 0.301), indicating a late, age-related change in the extracellular environment. These changes could lead to decreased conduction velocity or complete fiber dysfunction. A number of these changes resembled those of aged human peripheral nerves already examined.

Aging↗

PubMed2003

Adenosine extracellular levels in human brain gliomas: an intraoperative microdialysis study.

Adenosine present in human brain glioma extracellular spaces is a marker of astrocyte purine metabolism. In this study, we evaluated adenosine levels in the extracellular fluid of 21 human gliomas of high-grade malignancy using brain microdialysis techniques coupled to high-performance liquid chromatography. The adenosine concentration (mean+/-SEM) within the control tissue was 2.99+/-0.37 microM and in the tumour tissue 1.56+/-0.46 microM. The reduction was statistically significant. It is concluded that the adenosine concentrations reached in the tumour tissue are sufficient to stimulate all adenosine receptor subtypes, suppress local anti-tumour immune responses and affect glial and endothelial cell proliferation.

Adenosine↗

PubMed2000

Extracellular adenosine 5'-ATP-induced calcium signaling in isolated vestibular ganglion cells of the guinea pig.

Extracellular adenosine 5'-triphosphate (ATP)-induced intracellular calcium concentration ([Ca2+]i) changes in acutely isolated vestibular ganglion cells (VGCs) of the guinea pig were investigated using the Ca2+ -sensitive dye Fura-2. Extracellular ATP induced an increase in [Ca2+]i in VGCs in a dose-dependent manner. ATP induced an increase in [Ca2+]i even in the absence of extracellular Ca2+ (1 mM Ethylene Glycol-bis (beta-aminoethyl Ether) N,N,N',N'-Tetraacetic Acid (EGTA)), thus suggesting that ATP induces Ca2+ release from the intracellular stores. The P2-receptor antagonists suramin and reactive blue 2 inhibited the ATP-induced [Ca2+]i increase in a dose-dependent manner. The P1-receptor agonist adenosine did not induce any changes in [Ca2+]i. These results suggest that VGCs may possess a P2-purinergic receptor but not a P1-purinergic receptor. La3+, a receptor-mediated calcium channel blocker, inhibited the ATP-induced [Ca2+]i increase but, in contrast, nifedipine, a L-type calcium channel blocker, did not. These results suggest that ATP induces both a Ca2+ -release from the intracellular stores and a Ca2+ influx from the extracellular space through La3+ -sensitive and nifedipine-insensitive Ca2+ channels in VGCs. Our results also suggest that extracellular ATP may act as a neurotransmitter or neuromodulator of the vestibular peripheral system in the guinea pig.

Adenosine Triphosphate↗

PubMed1994

Accumulation of exogenous 45Ca after middle cerebral artery occlusion in rats.

The distribution of exogenous 45Ca in the focal ischemia rat model (middle cerebral artery occlusion) was studied using 45Ca autoradiography. High 45Ca accumulations were observed in the frontal cortex and caudate-putamen corresponding with morphological damage shown by HE staining. Regional 45Ca concentrations were calculated from the optical density on the 45Ca autoradiograms. Rapid uptake of 45Ca in the ischemic brain occurred during the first 5 hours, and continued more slowly between 5 and 24 hours after ischemia. The area of 45Ca accumulation was also expanded between 5 and 24 hours. An area of low 45Ca concentration around the area of high accumulation developed 5 hours after ischemia, which presumably accumulated 45Ca between 5 and 24 hours after ischemia. The lower concentration of 45Ca in the periphery of ischemia may result from: 1) a decrease in the total amount of calcium due to narrowing of extracellular space accompanied by cytotoxic edema, and 2) delayed accumulation of exogenous 45Ca due to reduced clearance of extracellular fluid.

Animals↗

PubMed1995

Persistent subcutaneous nodules in patients hyposensitized with aluminum-containing allergen extracts.

BACKGROUND: The development of persistent nodules that cause pain and itching at a vaccination or hyposensitization injection site is a rare event. These lesions have been mainly attributed to a hypersensitivity reaction to aluminum hydroxide, which is used as an absorbing agent in many vaccines and hyposensitization preparations. Patch tests with standard antigens and aluminum compounds and histopathologic and ultrastructural studies were performed on 10 patients with persistent subcutaneous nodules on the upper part of their arms after injection of aluminum-adsorbed dust and/or pollen extracts. OBSERVATIONS: The nodules appeared 1 month to 6.5 years after injections. The results of patch tests with 2% aluminum chloride were positive in five patients. Histopathologic examination revealed two different patterns: some biopsy specimens (from lesions of less than 9 months' duration) showed a pure foreign body histiocytic reaction characterized by extracellular amorphous dermal basophilic deposits with a histiocytic-macrophagic reaction; others showed a delayed hypersensitivity granulomatous reaction in association with an histiocytic foreign body response. The lesions were characterized by a unifocal or multifocal unencapsulated granulomatous reaction in the deep dermis and/or subcutaneous tissue. Eosinophilic necrotic areas surrounded by dense fibrous bands and a massive inflammatory infiltrate (lymphoid follicles, large histiocytic cells, abundant eosinophils, and some plasma cells) were observed. A granular basophilic material in extracellular spaces and within the cytoplasm of some histiocytes was also noted. Electron microscopic studies revealed intracytoplasmic and extracellular deposits of a fibrillar electron-dense material. CONCLUSIONS: Persistent subcutaneous nodules that develop after the administration of aluminum-containing preparations may show two characteristic histopathologic patterns. A pure histiocytic foreign body reaction was observed in early lesions, and a delayed hypersensitivity granulomatous reaction was seen in older lesions. No relationship between histopathologic pattern and patch test results was observed. Aluminum-free preparations should be used in patients in whom these nodules develop.

Adsorption↗

PubMed1983

Electron microscopic serial section analysis of nodes of Ranvier in lumbar spinal roots of the cat: a morphometric study of nodal compartments in fibres of different sizes.

Serially sectioned nodes of Ranvier from nerve fibres 2-20 micron in diameter of feline ventral and dorsal spinal roots were examined electron microscopically, reconstructed to scale and analysed morphometrically. The assumed 'fresh-state' value of several structural variables, considered to be of functional significance, were calculated by the use of compensation factors. The compensated data were plotted against fibre and axon diameters. It was calculated that the membranous area of the 'fresh-state' nodal axon segment increased more or less exponentially from less than 5 micron2 to 30 micron2 with increasing fibre diameter (D). Most variables associated with the nodal gap and the Schwann cell initially increased rapidly with D and then levelled out or even decreased in fibres with a D value greater than 8-12 micron. The area open for communication between the nodal axolemma and the endoneurial space was 30-100 times smaller than the membrane area of the nodal axolemma. The volume of the extracellular space in the nodal gap, outside the nodal axolemma, increased linearly from less than 0.1 micron3 to about 0.6 micron3 with increasing fibre size. The Schwann cell membrane area facing the nodal gap outnumbered the membrane area of the nodal axon by 10-15 times in nerve fibres with a D value between 5 and 15 microns. Some functional implications of the 'fresh-state' nodal model are discussed.

Animals↗

PubMed1988

Secretion of newly synthesized proteins into the extracellular fluid of the rabbit hippocampus.

Extracellular proteins were collected continuously from the hippocampus of the rabbit by slow perfusion of a protein-permeable thin tubing. After pulse labelling with radioactive amino acids via the tubing a rapid appearance of labelled proteins was seen in the extracellular fluid. The half-life of the proteins in the extracellular space varied between 15 min and 2 h. Protein fractions were separated with gel electrophoresis or HPLC. The extracellular fraction differed distinctly from intracellular proteins with respect to the labelling pattern.

Animals↗

PubMed1984

Control of extracellular potassium levels by retinal glial cell K+ siphoning.

Efflux of K+ from dissociated salamander Müller cells was measured with ion-selective microelectrodes. When the distal end of an isolated cell was exposed to high concentrations of extracellular K+, efflux occurred primarily from the endfoot, a cell process previously shown to contain most of the K+ conductance of the cell membrane. Computer simulations of K+ dynamics in the retina indicate that shunting ions through the Müller cell endfoot process is more effective in clearing local increases in extracellular K+ from the retina than is diffusion through extracellular space.

Ambystoma↗

PubMed1972

A compartmental analysis of 24 Na kinetics in rat cerebrum, sciatic nerve and cerebrospinal fluid.

1. The kinetics of (24)Na exchange in rat cerebrospinal fluid, brain and sciatic nerve were studied during steady-state plasma isotope conditions. The entrance of (24)Na into brain and sciatic nerve was found to fit a three compartmental model. Using newly derived equations for compartmental analysis, the first compartment (24)Na space of brain was found to be about 22% and the second compartment space about 6%; the corresponding sciatic nerve spaces were 26 and 11%, respectively. In the discussion, arguments are presented that suggest that the first compartment is the extracellular space (ECS) and the second compartment is the intracellular space. The t((1/2)) for compartmental equilibration between plasma and brain ECS was 2 hr and from ECS to intracellular space 14 hr; in nerve the respective t((1/2))'s were (2/3) hr and 10 hr.2. A third compartment of 18% was found in sciatic nerve which was interpreted to be anatomically confined to the epi- and perineurium; in brain the corresponding space was 2% and was felt to be anatomically and kinetically an inactive closed compartment in parallel with the blood.3. There were two t((1/2))'s for plasma-c.s.f. (24)Na equilibration of less than 10 min and 2 hr.4. Profiles of brain, from cortex to ventricle, at 6 min, demonstrated a gradient of decreasing (24)Na radioactivity from ventricle to cortex.

Animals↗

PubMed1994

Desynchronization of epileptiform activity by extracellular current pulses in rat hippocampal slices.

1. A single cathodic current pulse applied in the somatic CA1 region of the hippocampus was found to induce a large decrease in the amplitude of the population spike. 2. Intracellular recordings showed intense cellular firing suggesting the amplitude decrease could not be attributed to a decrease in neuronal firing. 3. Simultaneous intracellular and extracellular potentials were recorded to analyse the synchronization of neuronal firing in the CA1 region. Action potentials were synchronized with the first population spike but this synchronization decreased with subsequent spikes. Histograms of the phase of the action potentials displayed a normal distribution. 4. Histograms of the phases of the action potentials following the application of the 'singular stimulus' (one producing a singular response) revealed a uniform distribution of the phases suggesting that the neuronal population was desynchronized. 5. This desynchronization effect of the singular stimulus was verified by double intracellular recordings. The simultaneous firing of two neurons could be desynchronized by the application of the singular stimulus. 6. These findings indicate that it is possible to desynchronize a neuronal pool with the application of a single current pulse. In addition, the results show that it is possible for a neuronal population to fire a large number of action potentials with no resulting evoked potentials in the extracellular space.

Action Potentials↗

PubMed1998

Ictal epileptiform activity in the CA3 region of hippocampal slices produced by pilocarpine.

Pilocarpine, a muscarinic agonist, produces status epilepticus that is associated with the later development of chronic recurrent seizures. When applied to rat hippocampal slices, pilocarpine (10 microM) produced brief (<200 ms) epileptiform discharges that resembled interictal activity that occurs between seizures, as well as more prolonged synchronous neuronal activation that lasted seconds (3-20 s), and was comparable to ictal or seizures-like discharges. We assessed the factors that favored ictal patterns of activity and determined the biophysical properties of the ictal discharge. The probability of observing ictal discharges was increased when extracellular potassium ([K+]o) was increased from 5 to 7.5 mM. Raising [K+]o to 10 mM resulted in loss of ictal patterns and, in 20 of 34 slices, desynchronization of epileptiform activity. Making the artificial cerebrospinal fluid (ACSF) hyposmotic favored ictal discharges at 5 mM [K+]o, but shifted 7.5 mM [K+]o ACSF patterns to interictal discharges or desynchronized activity. Conversely, increasing osmolality suppressed ictal patterns. The pilocarpine-induced ictal discharges were blocked by atropine (1 microM, n = 5), a muscarinic antagonist, and pirenzepine (1 microM, n = 6), a selective M1 receptor antagonist. Kainate/alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid receptor blockade stopped all epileptiform activity (n = 8). The N-methyl--aspartate antagonist ,-2-amino-5-phosphonovaleric acid (100 microM, n = 34) did not change the pattern of epileptiform activity but significantly increased the rate of interictal discharges and prolonged the duration of ictal discharges. The ictal discharge was characterized intracellularly by a depolarization that was associated with action potential generation and persisted as a membrane oscillation of 4-10 Hz. The ictal oscillations reversed in polarity at -22.7 +/- 2.2 mV (n = 11) with current-clamp recordings and -20.9 +/- 3.1 mV (n = 7) with voltage-clamp recordings. The reversal potential of the ictal discharge in the presence of the gamma-aminobutyric acid-A blocker bicuculline (10 microM, n = 6) was -2.2 +/- 2.6 mV and was significantly different from that measured without bicuculline. Bicuculline added to 7.5 mM [K+]o and 10 microM pilocarpine did not cause epileptiform activity to change pattern but significantly increased the rate of interictal discharges and prolonged the ictal discharge duration. Both synaptic and nonsynaptic mechanisms are important for the generation of ictal patterns of epileptiform activity. Although the synchronous epileptiform activity produced by pilocarpine required fast glutamate-mediated synaptic transmission, the transition from an interictal to ictal pattern of activity depended on [K+]o and could be influenced by extracellular space.

2-Amino-5-phosphonovalerate↗

PubMed1987

Phenylphosphonate: a 31P-NMR indicator of extracellular pH and volume in the isolated perfused rabbit bladder.

31P-NMR has been used extensively to estimate intracellular pH. It also can be used to measure extracellular pH and volume when an NMR-detectable extracellular phosphorous probe is used. Phosphonic acids have been suggested as useful 31P-NMR extracellular markers. The present study was designed to assess the utility of phenylphosphonic acid (PPA) as a 31P-NMR extracellular marker in perfused smooth muscle. Rabbit bladder strips were exposed to PPA concentrations of 1-20 mM. Tension development in response to maximal carbachol challenges (10 microM) was independent of PPA concentration. Addition of PPA (6 mM) to the perfusate supplying the isolated resting rabbit bladder had no effect on 31P-NMR-detectable phosphatic compounds. PPA's resonance frequency was distinctly downfield from endogenous phosphates and demonstrated a pH-dependent chemical shift of +/- 1.12 ppm/pH unit over the range of 6.4 to 7.6 with a pK' of 7.09 at 23 degrees C. The time courses for washing PPA in and out of the resting bladder were best described by monotonic exponential growth (r = 0.972; n = 3) and decay (r = 0.972; n = 3) equations, respectively. Rate and time constants for PPA wash-in (0.039 +/- 0.004 min-1 and 25.7 +/- 2.3 minutes) and washout (0.038 +/- 0.000 min-1 and 26.3 +/- 0.0 minutes) were not significantly different. Using steady state PPA and ATP peak intensities and concentrations, an extracellular-to-intracellular ratio was calculated to be 0.31 +/- 0.03 (n = 3). These data indicate that PPA remains distributed exclusively in the extracellular spaces.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1988

Electric impedance measurements in quadriplegia.

Previous experimental studies have shown that measurements of electric impedance of the whole human body relate to body fluid volumes. Measurements using two properly selected current frequencies have been purported to be useful in determining the sizes of extracellular and total body fluids. In this study, measurements of whole body impedance were performed on 20 healthy able-bodied subjects and 20 patients with quadriplegia at the frequencies of 1.5kHz and 150kHz. Differences found between the two groups suggest that the ratio of interstitial body fluids to total body fluids in quadriplegic patients is higher than in healthy subjects. This finding is consistent with previous body composition studies conducted on quadriplegic patients using a radioisotope technique, which have suggested a relative increase of extracellular space.

Adult↗