Determination of the content of nonfilterable cells in erythrocyte suspensions as a function of the medium osmolality

Lisovskaya, I. L.; Shurkhina, E. S.; Nesterenko, V. M.; Rozenberg, Julian M.; Ataullakhanov, F. I.

doi:10.1016/s0006-355x(99)80004-3

Cited by 12 publications

(13 citation statements)

References 21 publications

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“…2052194 of the Russian Federation) developed and made at Laboratory of Physical Biochemistry of Hematological Center. This hemorheometer measures the time of fi ltration of a fi xed volume of fl uid (250 μl) under a pressure of 60 mm H 2 O [2,8]. Polycarbonate nuclepore fi lters were 7 μ thick with the mean diameter of pores 3 μ, 3.5-3.8×10 5 pores per working surface.…”

Section: Methodsmentioning

confidence: 99%

“…Erythrocyte capacity to pass through narrow fi lter pores with a diameter (3 μ) lesser than the diameter of a resting erythrocyte (8 μ) is explained by its excessive surface in comparison with a sphere of the same volume [2,8]. Due to this, the cell can reversibly change its shape (be deformed) if necessary and penetrate through narrow holes (capillaries, fi lter pores, etc.).…”

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confidence: 99%

“…The erythrocyte volume increases and the ratio of cell surface to cell volume decreases. At a certain osmolarity value depending on the initial geometrical parameters of the cell, the erythrocyte turns into a sphere and no longer passes through the fi lter pores [2,8].…”

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confidence: 99%

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Filtration Method for Studies of the Kinetics of Hypo-Osmotic Pore Closure in Erythrocyte

et al. 2010

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Filterability of erythrocytes through small (3 μ) pores decreases with decreasing osmolarity of suspension medium because of hypo-osmotic swelling of cells. After appearance of lytic pores, erythrocyte filterability increases for some time, while after recovery of membrane integrity it decreases again. We suggest filtration method for studies of the kinetics of hypo-osmotic lytic pores closure. The dynamics of changes in erythrocyte filterability was studied in 2 patients with paroxysmal nocturnal hemoglobinuria and 6 donors (Ht 0.01%, Na phosphate buffer 5 mM, pH 7.4, 35 mOsm, 24°C). The method can be used for studies of erythrocyte membrane characteristics in various diseases and for evaluation of the membranotropic effects of drugs, infusion media, hemolysins, ethanol, etc.

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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Filtration Method for Studies of the Kinetics of Hypo-Osmotic Pore Closure in Erythrocyte

et al. 2010

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“…The EOR is determined from the amount of hemoglobin entering the liquid owing to the destruction of erythrocytes. A filtration-osmotic method has been developed [3][4][5] which combines the use of membrane filters and solutions with different osmotic concentrations. As in the earlier method, sample preparation includes centrifuging.…”

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Light scattering by erythrocytes under conditions of osmotic imbalance in a cell-surrounding medium system

Naumenko

2009

J Appl Spectrosc

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A model for changes in the sizes of erythrocytes under conditions of osmotic imbalance in a hypoosmotic medium is proposed assuming free diffusion of water through the erythrocyte membrane. Equations relating the minimum and maximum osmotic resistances to the shape of an erythrocyte in an isotonic medium are obtained. A numerical simulation is used to determine how the optical attenuation coefficient varies as a result of the swelling and hemolysis of erythrocytes; this provides a foundation for a new optical method for determining the osmotic resistance of erythrocytes. This method does not require exerting a force on the erythrocytes, is distinguished by simplicity and high speed, and can be implemented using standard spectrophotometers.Introduction. Erythrocytes, their shape and rheological properties -deformability, ability to undergo aggregation, stability with respect to external factors -play an important role in exchange of oxygen, carbon dioxide, and metabolic products in tissues. Over the time they are viable, erythrocytes undergo various kinds of mechanical and chemical interactions which require that their membranes be durable, flexible, and elastic. The ability of erythrocytes to withstand the action of external factors is characterized by the concept of their resistance. As the resistance decreases to a minimum, hemolysis sets in; hemolysis is the failure of the membrane of erythrocytes, accompanied by the escape of hemoglobin into the blood plasma. Depending on the nature of the destructive agent, a distinction is made between osmotic, mechanical, chemical, and thermal hemolysis. Hemolysis can also be caused by UV, ionizing radiation, and x-rays. It is known that, regardless of the type of hemolysis, the breakdown of erythrocytes is preceded by a change in their shape to spherical and swelling to a certain limit. Given this, specialists say that the hemolysis process leads to an excess intracellular osmotic concentration above the external pressure and to rupture of the membrane caused by the internal osmotic pressure.The osmotic resistance of erythrocytes refers to their stability in hypotonic solutions (solutions with an osmotic concentration below that for osmotic equilibrium). The most widespread method of measuring the erythrocyte osmotic resistance (EOR) under clinical conditions is to place the erythrocytes in hypotonic solutions with different concentrations of NaCl [1,2]. After a certain amount of time the samples are centrifuged and the resulting liquid is analyzed photometrically. The EOR is determined from the amount of hemoglobin entering the liquid owing to the destruction of erythrocytes. A filtration-osmotic method has been developed [3][4][5] which combines the use of membrane filters and solutions with different osmotic concentrations. As in the earlier method, sample preparation includes centrifuging. The samples pass through a filter with an average pore size smaller than the dimensions of the erythrocytes. The EOR is determined from the rate at which a fixed volume of suspensio...

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“…Furthermore, the effect of nonfilterable cells also cannot be excluded. 15 The suspensions' hematocrit values were correct; if not, this would have been a huge source of error in determining the filtration parameters. Furthermore, it is important to remark that in the single suspensions, the mean corpuscular volume did not change considerably.…”

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Measurement of erythrocyte deformability and methodological adaptation for small‐animal microsurgical models

et al. 2006

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Measuring erythrocyte deformability with bulk filtrometers is common, but there are problems with standardization and sample quantity in small laboratory animals. In this study, blood was drawn from mice, rats, and dogs, and then erythrocyte suspensions at 1-5% hematocrit were prepared. Bulk filtrometers with a 5- or 3-microm pore-diameter filter were used to determine the initial relative filtration rate (IRFR) and relative cell transit time (RCTT). Hematological composition was also tested. In all species, the highest IRFR values occurred at 1% hematocrit, and the lowest values at 5%. In mice and rats, RCTT values were lowest at 1% and highest at 5% using a 5-microm filter, whereas in dogs, RCTT was elevated at 1-2%. Using a 3-microm filter, each species showed the same phenomena under 3%. RCTT values may become distorted above a certain cell-size/pore-size ratio. In mice and rats, 1% suspension is applicable for bulk filtrometry, but only using 5-microm filters.

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Determination of the content of nonfilterable cells in erythrocyte suspensions as a function of the medium osmolality

Cited by 12 publications

References 21 publications

Filtration Method for Studies of the Kinetics of Hypo-Osmotic Pore Closure in Erythrocyte

Filtration Method for Studies of the Kinetics of Hypo-Osmotic Pore Closure in Erythrocyte

Light scattering by erythrocytes under conditions of osmotic imbalance in a cell-surrounding medium system

Measurement of erythrocyte deformability and methodological adaptation for small‐animal microsurgical models

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