The Renal Excretion of Hemoglobin: Regulatory Mechanisms and the Differential Excretion of Free and Protein-Bound Hemoglobin*†

Lathem, Willoughby

doi:10.1172/jci103843

Cited by 91 publications

(34 citation statements)

References 12 publications

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“…Hemoglobin was administered intravenously in either single or multiple injections to 13 healthy, fasting adult subjects who showed no evidence of cardiovascular, infectious, hematologic or renal disease. Hemoglobin was prepared from autogenous blood which was withdrawn in heparinized syringes and hemolyzed by the addition of 2.5 volumes of distilled water to each volume of whole blood.…”

Section: Methodsmentioning

confidence: 99%

The Distribution of Extracorpuscular Hemoglobin in Circulating Plasma*†

Lathem¹,

Worley²

1959

J. Clin. Invest.

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Section: Methodsmentioning

confidence: 99%

The Distribution of Extracorpuscular Hemoglobin in Circulating Plasma*†

Lathem¹,

Worley²

1959

J. Clin. Invest.

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“…Conventionally, inferences regarding hemoglobin reabsorption have been made in one of two ways: on the basis of a demonstrable threshold level of excretion (5,7) or by demonstrating a difference between the rate of filtration of hemoglobin and its rate of excretion (7,8). However, neither of these methods has proved satisfactory or conclusive in recent studies in man (9) and the dog ( 10), in which it has been shown that the threshold results largely from plasma protein binding and that the differences between rates of filtration and excretion are small or negligible.…”

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confidence: 99%

The Demonstration and Localization of Renal Tubular Reabsorption of Hemoglobin by Stop Flow Analysis *

Lathem¹,

Davis²,

Zweig³

et al. 1960

J. Clin. Invest.

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The evidence that extracorpuscular hemoglobin is reabsorbed by renal tubular cells after filtration through the glomerulus is based principally upon morphological studies. In a variety of animal species hemoglobin or its metabolic derivatives can be demonstrated in tubular epithelium during or following spontaneous or induced hemoglobinemia (1-6).Although these histological observations suggest that hemoglobin enters tubular cells as a result of reabsorptive activity, attempts to demonstrate such activity in the intact animal by physiological methods have been equivocal. Conventionally, inferences regarding hemoglobin reabsorption have been made in one of two ways: on the basis of a demonstrable threshold level of excretion (5, 7) or by demonstrating a difference between the rate of filtration of hemoglobin and its rate of excretion (7,8). However, neither of these methods has proved satisfactory or conclusive in recent studies in man (9) and the dog ( 10), in which it has been shown that the threshold results largely from plasma protein binding and that the differences between rates of filtration and excretion are small or negligible.In the present study an attempt was made to demonstrate reabsorptive activity in the intact animal (the dog) in another way: by the use of stop flow analysis (11). This method is based on the principle that, when urinary outflow is suddenly obstructed, the composition of urine trapped in metabolically active tubular segments is altered, owing to continued tubular reabsorptive or secretory activity, the effect of such alterations becoming apparent on restoration of * Supported by a grant-in-aid (A-1971) from the

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“…Die bisherigen Veröffentlichungen zur Hämoglöbinbestimmung basieren überwiegend auf der Anwendung der PeroxidaseReaktion. Zumeist wird der quantitative Nachweis mit dem Elektrophorese^Verfahren kombiniert, was jedoch eine geeignete apparative Einrichtung voraussetzt (3)(4)(5)(6)(7)(8)(9).…”

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Eine einfache Methode zur Bestimmung von freiem und proteingebundenem Hämoglobin im Plasma

Weichert¹,

Schulz²

1974

Clinical Chemistry and Laboratory Medicine

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Im Rahmen experimenteller Untersuchungen zur glomerulokapillären Permeabilität der Hundeniere für Hämoglobin stellte sich das Problem eines einfachen Nachweises von freiem (Hbf) und proteingebundenem (Hb^) Hämoglobin im Plasma. Nachdem in Vorversuchen die Aussalzung des proteingebundenen Hämoglobins gesichert werden konnte, wurde folgendes Verfahren angewandt: Hämoglobinhaltiges Plasma wurde mit Natriumsulfit (280 g/l) im Volumen-Verhältnis 1:11 für 3 h bei 37°C inkubiert. Die Proben wurden mit Diäthyläther überschichtet, kurz geschüttelt und für 10 min bei 2750 g zentrifugiert. Der Überstand wurde abgesaugt. Der Hämoglobingehalt der wäßrigen Phase (Hbf) sowie die Ausgangskonzentration des Plasmas an Hämoglobin (Hb tota j) wurden mit der Hämiglobin-Cyanid-Methode gemessen. Die Konzentration des proteingebundenen Hämoglobins (Hbjj) ergibt sich dann unter Berück-sichtigung des Verdünnungsfaktors aus der Beziehung Hb^ = Hb tota j -Hbf. Die methodischen Grenzen des Verfahrens werden diskutiert A simple method for the determination of free and protein-bound haemoglobin in plasmaIn the course of experiments on the glomerulo-capillary permeability of dog kidney with respect to haemoglobin, a simple method was required for the measurement of free (Hbf) and protein-bound (Hb^) haemoglobin in the plasma. On the basis of the observation that the protein-bound haemoglobin can be separated by salting out, the following method was developed: Plasma containing haemoglobin (Hb tota i) was incubated for 3 hr at 37° C with sodium sulphite solution (280 g/1) in the volume ratio 1:11 respectively. After the addition of ether, samples were shaken briefly by hand, then centrifuged (2750 #,10 min). The upper layer with the precipitated protein was removed by suction at the pump, and the concentrations of haemoglobin in the aqueous layer (Hbf) and in the starting plasma (Hb to t a j) were measured by the haemiglobin-cyanide method. Taking dilution factors into account, the concentration of protein-bound haemoglobin is defined as Hb b = Hb tota i -Hbf. Limitations of the method are discussed.

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The Renal Excretion of Hemoglobin: Regulatory Mechanisms and the Differential Excretion of Free and Protein-Bound Hemoglobin*†

Cited by 91 publications

References 12 publications

The Distribution of Extracorpuscular Hemoglobin in Circulating Plasma*†

The Distribution of Extracorpuscular Hemoglobin in Circulating Plasma*†

The Demonstration and Localization of Renal Tubular Reabsorption of Hemoglobin by Stop Flow Analysis *

Eine einfache Methode zur Bestimmung von freiem und proteingebundenem Hämoglobin im Plasma

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