A method using 3-O-methyl-D-glucose and phloretin for the determination of intracellular water space of cells in monolayer culture

One approach to platelet sizing is to measure the intracellular water space of platelets with 3H-H2O since the % water content of platelets remains constant in states with different platelet sizes. Fresh citrated blood was centrifuged for 10 min at 150 'g' to obtain PRP. Aliquots of PRP were briefly incubated with either 3H-H2O or 14C-sucrose then layered over 0.3 ml dibutylphthalate and spun 4 min at 8000 'g'. The cell pellet was solubilized and counted to enable spaces to be calculated. The extracellular (sucrose) space was subtracted from the total water space of the pellet to give a mean intracellular water space of 0.56 ± 0.12 μ1/108 platelets (n =19). Assuming a water content of 7 5% and a density of 1.04, the mean platelet volume for normal subjects is 7.2 fl. Gel-filtration of platelets (GFP) on Sepharose-2B reduced their mean water space by 0.12 μl/108 platelets. However the amount of shrinkage on gel-filtration depended on the initial water space of the platelets in PRP and there was a linear relation between these two variables (r = 0.82). Shrinkage was 40% for an initial platelet water space of 0.70 μl/108 platelets but there was almost no shrinkage below a water space of 0.40 μl/108 platelets. Recovery of platelets from each column averaged 8 0% and showed no relation to the reduction in the mean cell water space. The lower water space of GFP may indicate a reduction in mean cell volume due to gel-filtration.

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Use of 3-O-methyI-D-glucose and phloretin to estimate the intracellular water space of mouse peritoneal macrophage monolayers

Canadian Journal of Physiology and Pharmacology ◽

10.1139/y82-010 ◽

1982 ◽

Vol 60 (1) ◽

pp. 76-79

Author(s):

Douglas B. Lowrie

Keyword(s):

Glucose Uptake ◽

Peritoneal Macrophage ◽

Intracellular Water ◽

Mouse Peritoneal Macrophage ◽

Facilitated Diffusion ◽

Water Space ◽

Glucose Carrier

Macrophages took up 3-O-methyl-D-glucose rapidly by facilitated diffusion using the glucose carrier and slowly by carrier-independent diffusion. Phloretin inhibited carrier-dependent but not carrier-independent diffusion. Estimates of intracellular water space based on 3-O-methyl-D-glucose uptake varied between 0.7 and 6.9 μL∙106 cells−1 for 2-week-old monolayers.

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Optical determination of intracellular water in apoptotic cells

The Journal of Physiology ◽

10.1113/jphysiol.2013.263228 ◽

2013 ◽

Vol 591 (23) ◽

pp. 5843-5849 ◽

Cited By ~ 16

Author(s):

Michael A. Model ◽

Ethan Schonbrun

Keyword(s):

Intracellular Water ◽

Apoptotic Cells ◽

Optical Determination

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Determination of Intracellular Water by Multifrequency Bioelectrical Impedance

Annals of Nutrition and Metabolism ◽

10.1159/000177860 ◽

1995 ◽

Vol 39 (3) ◽

pp. 177-184 ◽

Cited By ~ 27

Author(s):

A. De Lorenzo ◽

N. Candeloro ◽

A. Andreoli ◽

P. Deurenberg

Keyword(s):

Bioelectrical Impedance ◽

Intracellular Water

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The effect of route of dosing and method of estimation of tritiated water space on the determination of total body water and the prediction of body fat in sheep

The Journal of Agricultural Science ◽

10.1017/s0021859600050279 ◽

1974 ◽

Vol 82 (1) ◽

pp. 105-112 ◽

Cited By ~ 16

Author(s):

B. S. W. Smith ◽

A. R. Sykes

Keyword(s):

Body Weight ◽

Body Fat ◽

Total Body Water ◽

Specific Activity ◽

Tritiated Water ◽

Multiple Correlation Coefficient ◽

Body Water ◽

Total Body ◽

Water Space

SUMMARYEight mature female sheep were offered a ration which maintained body weight constant during a 20-week period. During the final 10 weeks a comparison was made in each animal of the pattern of equilibration and urinary losses of tritiated water during 8 h after dosing by four different routes. These were intravenous, intraperitoneal, intraruminal and a combination of the intraperitoneal and intraruminal routes. Tritiated water spaces were calculated from (a) the 8-h plasma specific activity and (b) by extrapolation to zero time of the plasma specific activities during the 7 days after injection. At the end of the experiment the fat and water contents of the bodies of the sheep were determined directly.Complete equilibration of tritiated water between plasma and rumen water was not achieved in all animals 8 h after intravenous or intraperitoneal injection but was when the rumen was primed by the combination of intraperitoneal and intraruminal dosing. After intraruminal dosing equilibration was not achieved in any animal within 8 h of dosing.Urinary losses of marker were lower after intraruminal dosing but otherwise averaged 4–5 % of the dose/1 urine. This was equivalent to 0·3–6·7% of the dose for individual sheep.Errors resulting from incomplete equilibration and urinary loss of marker did not influence the efficiency of prediction of total body water from tritiated water space. The multiple correlation coefficient relating body fat with empty body weight and its water content was very high (r = 0·99). Errors introduced into this relationship by the inclusion of gut water in the prediction equations were apparently of a similar magnitude to those resulting from the errors in the estimation of tritiated water space.The extrapolation method for the determination of tritiated water space was shown to have the same accuracy as equilibration techniques under these controlled dietary conditions.

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Rates of efflux and intracellular concentrations of potassium, sodium and chloride ions in isolated fat-cells from the rat

Biochemical Journal ◽

10.1042/bj1150865 ◽

1969 ◽

Vol 115 (5) ◽

pp. 865-871 ◽

Cited By ~ 21

Author(s):

M. C. Perry ◽

C. N. Hales

Keyword(s):

Atomic Absorption Spectrophotometry ◽

Chloride Ions ◽

Intracellular Water ◽

Epididymal Adipose Tissue ◽

Fat Cells ◽

Isolated Fat Cells ◽

First Order ◽

Inulin Space ◽

Intracellular Concentrations ◽

Water Space

1. The metabolism of K+, Na+ and Cl− has been investigated in isolated fat-cells prepared from the epididymal adipose tissue of rats. 2. Methods are described for measuring the intracellular water space, the rates of loss of intracellular 42K+, 22Na+ and 36Cl− and the intracellular concentrations of K+, Na+ and Cl− in isolated fat-cells. 3. The intracellular water space, measured as the [3H]water space minus the [carboxylic acid−14C]inulin space, was 3·93±0·38μl./100mg. cell dry wt. 4. The first-order rate constants for radioisotope effluxes from isolated fat-cells were 0·029min.−1 for 42K+, 0·245min.−1 for 22Na+ and 0·158min.−1 for 36Cl−. 5. The intracellular concentrations of K+, Na+ and Cl− were 146m-equiv./l., 18·6±2·9m-equiv./l. and 43±2·4m-equiv./l. respectively. 6. The total intracellular K+ content of isolated fat-cells was determined by atomic-absorption spectrophotometry to confirm the value obtained from the radioisotope-efflux data. 7. The ion effluxes from isolated fat-cells were: K+, 1·5pmoles/cm.2/sec., Na+, 1·6pmoles/cm.2/sec., and Cl−, 2·4pmoles/cm.2/sec. 8. The membrane potential of isolated fat-cells calculated from the Cl− distribution ratio was −28·7mv.

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Relaxometric investigations addressing the determination of intracellular water lifetime: a novel tumour biomarker of general applicability

Molecular Physics ◽

10.1080/00268976.2018.1527045 ◽

2018 ◽

Vol 117 (7-8) ◽

pp. 968-974 ◽

Cited By ~ 5

Author(s):

Maria Rosaria Ruggiero ◽

Simona Baroni ◽

Silvio Aime ◽

Simonetta Geninatti Crich

Keyword(s):

Intracellular Water ◽

General Applicability

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Этимология русского диалектного слова анчутка

Acta Baltico-Slavica ◽

10.11649/abs.2011.010 ◽

2015 ◽

Vol 35 ◽

pp. 131-141

Author(s):

Rolandas Kregždys

Keyword(s):

Russian Word ◽

Areal Distribution ◽

Poor Person ◽

Water Space

The etymology of the Russian dialectal lexeme aнчуткаThe article deals with the etymological analysis of Russ. dial. анчýтка 'devil, demon; Antichrist; hobgoblin, water-sprite, familiar, sylvan; trollop; poor person; pickle; curse'. The precondition is made, that it reflects multipartite derivative: prefix Russ. на-+ radix Russ. чуд- [with secondary -m- (phonetic recording) instead of etymological -д-] + suff. -ка → *начудкa. Being based on the made lexeme analysis, Russ. dial. анчýтка is explained as primordially Russian (!!!) word, which contrarily V. N. Toporov's hypothesis by no means could be compared with the water space birds names. Due to such decision it can not be explained as Baltism either. By virtue of it, determination of the Protobaltic tribes presumptive borders of their residing territory in Russia, according to areal distribution of the word, is not possible.

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Regulation of liver cell volume and proteolysis by glucagon and insulin

Biochemical Journal ◽

10.1042/bj2780771 ◽

1991 ◽

Vol 278 (3) ◽

pp. 771-777 ◽

Cited By ~ 51

Author(s):

S Vom Dahl ◽

C Hallbrucker ◽

F Lang ◽

W Gerok ◽

D Häussinger

Keyword(s):

Cell Volume ◽

Liver Cell ◽

Intracellular Water ◽

Cell Swelling ◽

Isolated Perfused Rat Liver ◽

Cell Shrinkage ◽

Glucagon And Insulin ◽

Opposing Effects ◽

Water Space

The effects of insulin and glucagon on liver cell volume and proteolysis were studied in isolated perfused rat liver. The rate of proteolysis was assessed as [3H]leucine release from single-pass-perfused livers from rats which had been prelabelled in vivo by intraperitoneal injection of [3H]leucine. The intracellular water space was determined from the wash-out profiles of simultaneously added [3H]inulin and [14C]urea. In normo-osmotic (305 mosM) control perfusions the intracellular water space was 548 +/- 10 microliters/g wet mass (n = 44) and was increased by 16.5 +/- 2.6% (n = 6), i.e. by 85 +/- 14 microliters/g, after hypoosmotic exposure (225 mosM). Glucagon (0.1 microM) decreased the intracellular water space by 17 +/- 4% (n = 4), whereas insulin (35 nM) increased the intracellular water space by 9.3 +/- 1.4% (n = 15). Also, in isolated rat hepatocyte suspensions insulin (100 nM) caused cell swelling by 10.7 +/- 1.8% (n = 16), which was fully reversed by glucagon. In perfused liver, insulin-induced cell swelling was accompanied by a hepatic net K+ uptake (4.5 +/- 0.2 mumol/g) and an inhibition of proteolysis by 21 +/- 2% (n = 12); further addition of glucagon led to a net K+ release of 3.8 +/- 0.2 mumol/g (n = 7) and fully reversed the insulin effects on both cell volume and proteolysis. Similarly, insulin-induced cell swelling and inhibition of proteolysis were completely antagonized by hyperosmotic (385 mosM) cell shrinkage. Furthermore, cell swelling and inhibition of proteolysis after hypo-osmotic exposure or amino acid addition were reversed by glucagon-induced cell shrinkage. There was a close relationship between the extent of cell swelling and the inhibition of proteolysis, regardless of whether cell volume was modified by insulin, glucagon or aniso-osmotic exposure. The data show that glucagon and insulin are potent modulators of liver cell volume, at least in part by alterations of cellular K+ balance, and that their opposing effects on hepatic proteolysis can largely be explained by opposing effects on cell volume. It is hypothesized that hormone-induced alterations of cell volume may represent an important, not yet recognized, mechanism mediating hormonal effects on metabolism.

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