Ein mikrophotometrisches Verfahren zur Aufnahme der Sauerstoffbindungskurve von einzelnen Erythrocyten

Zusammenfassung Es wird ein Verfahren zur mikrophotometrischen Aufnahme von Sauerstoffbindungskurven beschrieben, mit dessen Hilfe es möglich ist, unmittelbar nacheinander sowohl von einzelnen Erythrocyten als auch einer größeren Zahl roter Blutkörperchen die O₂-Sättigung in Abhängigkeit vom O₂-Druck zu bestimmen. Außerdem kann mit dieser Methode der Hämoglobingehalt einzelner Erythrocyten ermittelt werden. Die von einzelnen Blutkörperchen erhaltenen Meßergebnisse waren gut reproduzierbar. Die erforderliche Blutmenge für eine Kammerbeschickung beträgt weniger als 1 l. Bestimmungen an 100 einzelnen Erythrocyten desselben Blutes zeigten beträchtliche Variationen des O₂-Bindungskurvenverlaufes. Es fand sich ein Mittelwert des Halbsättigungsdruckes von 25,4±4,6 Torr, wobei die Extremwerte 14 bzw. 47 Torr betrugen. Dieses Ergebnis zeigt, daß Sauerstoffbindungskurven, die mit den bisher üblichen Methoden gewonnen wurden, Mittelwertkurven darstellen. Der Hämoglobingehalt der untersuchten 100 Erythrocyten läßt eine Normalverteilung erkennen bei einem Mittelwert von 32,3·10^–12±6,1·10^–12 g pro Blutkörperchen.

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Summary A method for the microphotometric determination of the O₂-dissociation curve is decribed which allows us to investigate the relation between O₂-saturation and O₂-pressure of single erythrocytes as well as of a greater number of red cells immediately in subsequence. Moreover, by this method we can determine the hemoglobin content of single red cells. The results of the measurements received from single cells were well reproducable. The necessary quantity of blood for one charging of the chamber amounts to less than 1 l. Determinations of 100 single erythrocytes from the same blood showed considerable variations in the course of the O₂-dissociation curve. The mean value of O₂-pressure necessary for half saturation of the cells was 25.4±4.6 Torr. The extremes were 14 respectively 47 Torr. This result shows clearly that the O₂-dissociation curves received by usual methods till now represent curves of middle value. The hemoglobin content of the studied 100 erythrocytes shows a Gaussian distribution with a mean value of 32.3±6.1·10^–12 g per blood cell.

     

Aufnahme von Zink durch synchrone Zellen von Chlorella fusca

The uptake of labelled zinc into the interior of synchronous Chlorella fusca was measured at 30 °C in minimal and optimal conditions (dark/nitrogen or light/air, respectively). No saturation with Zn was reached during 10 hours. Uptake strongly depended on the stage of the cells during the development cycle. The rates of uptake per unit cell number, per unit cell volume and also per unit cell surface, reach maxima be forecell division, and thereafter strongly decrease. Clearly the transport system is built up during cell growth. The similarity of the dependence of the rates of uptake in minimal and optimal conditions on the stage of development suggests identity of the transport systems.     

Aufnahme von markiertem Zink durch Chlorella unterschiedlichen Zinkgehalts

Chlorella fusca was grown asynchronously for 2 weeks in media with different Zn concentrations up to 10^?3 m. Growth was optimal at 10^?5 m Zn. The Zn contents of the algae were followed by activation analysis. They increased with increasing Zn concentration in the media, and, except with the lowest Zn concentrations, with time. Further, the uptake of labelled Zn (⁶⁵Zn) from 5 · 10^?6 m solution by cells of known Zn content was measured at 30 °C during one day in minimum and optimum conditions (dark/nitrogen or light/air, respectively). The rate of uptake showed a maximum with algae grown in a medium with 10^?5 m Zn. However, algae exposed to a lethal Zn concentration (10^?3 m) also absorbed much Zn; this anomalous uptake is ascribed to a breakdown of the cell membrane as a permeability barrier.     

Zur Aufnahme von markierten Zellpartikeln durch Tumorzellen in vitro

Ohne ZusammenfassungDie Arbeit wurde mit Unterstützung der Deutschen Forschungsgemeinschaft durchgeführt.     

Die Ausscheidung von J131 durch das Bronchialsekret

Zusammenfassung An 9 laryngektomierten Patienten wurden 12 Untersuchungen über die Radiojodausscheidung in Sputum nach Vorbehandlung mit 9 g Kaliumjodid durchgeführt. Die J¹³¹-Ausscheidung der gesamten, während 46 Std gesammelten Sputummengen zeigte ein für die einzelnen Fälle uneinheitliches Aktivitätsmaximum, fiel aber in allen Fällen nach 24 Std ab. Von dem eingenommenen J¹³¹ fand sich im Sputum nur die sehr geringe Menge von 0,01–0,4% der Gesamtaktivität wieder.Herrn Prof. Dr.J. H. Müller, Zürich, Radiologische Abteilung und Histopathologisches Laboratorium der Universitäts-Frauenklinik, in Verehrung gewidmet.     

Aufnahme von Cobalt,Blei und Cadmium durch Bäckerhefe

The toxicity for and the uptake by Saccharomyces cerevisiae of the essential trace element Co²⁺ and the non essential elements Cd²⁺ and Pb²⁺ were compared. Inhibition of yeast growth is observed 4 hrs after addition of Co at concentrations higher than 10^?4 M. Cd is about 100 times more toxic than Co and inhibits growth immediately after addition. No toxicity of Pb is observed with concentrations up to 5 · 10^?4 M. For uptake experiments the radioactive isotopes ⁶⁰Co, ^115mCd and ²¹⁰Pb were used. The yeast samples were washed with carrier solution in order to remove the isotopes adsorbed only by the cell wall and measured by γ-scintillation counting. Co uptake by yeast is strongly dependent on glucose both under aerobic and anaerobic conditions. There is no efflux of Co once taken up. Dead yeast does not accumulate Co at all. Co already taken up is released from cells when killed, indicating that no irreversible binding in the cell interior takes place. Co transport follows biphasic saturation kinetics with the MICHAELIS constants K₁ = 10^?4 and K₂ = 8 · 10^?4 M and the maximal velocities ν¹ = 9.5 and ν₂ = 63 μ moles/g dry weight · hr. The Q₁₀ for Co uptake from 10^?4 M solution is 2.3. Uptake of Cd resembles that of Co. The absorption of Cd is also glucose-dependent, but independent of the presence of air, no Cd is taken up by dead cells, and Cd previously taken up is released during killing the cells. There is a considerable Pb uptake irrespective of the presence of glucose, and even by dead yeast. Killing of the yeast during uptake does not lead to loss of Pb. It is concluded that the essential trace element Co is accumulated by an energy-dependent, probably active, transport system in the membrane. Cd is similarly transported, probably because of its chemical similarity to Zn. In contrast, Pb appears to be taken up only by diffusion, but subsequently to be trapped by binding in the cell interior. In this way, the accumulation of Pb is similar in amount to those of Co or Cd although the mechanisms differ widely.     

Hemmung des Kaliumverlustes fluoridvergifteter Erythrocyten durch Komplexbildner

Summary Fluoride poisened (40 mequiv./l.) human red cells lose potassium at rates of about 25 mequiv./l. cells/hr. This rapid potassium loss is reduced to 1.6 to 2.0 mequiv./l. cells/hr. by the addition of ethylendiaminetetra acetic acid (EDTA) or citrate. Oxalate is ineffective.EDTA enters neither intact nor fluoride poisened erythrocytes.The action of the complexing agents is reversed by a surplus of Ca⁺⁺ or Mg⁺⁺.The drastic decrease of cellular ATP-content associated with fluoride poisoning of red cells is not inhibited by EDTA.It has been concluded that the slow potassium loss of 1.6 to 2.0 mequiv./l. cells/hr. is due to the absence of an energy supply for active cation transport, induced by the inhibition of glycolysis by fluoride; whereas the rapid potassium loss is caused by an additional or independent reaction of fluoride in the cell surface. This latter reaction seems to involve either the formation of a metal-fluoride-complex with ligands in the cell membrane or the activitation by Ca⁺⁺ or Mg⁺⁺ of abnormal metabolic reactions in the cell surface, which lead to a breakdown of the permeability barrier.

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Mit 3 Textabbildungen

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Herrn Professor Dr. Rudolf Mond zum 65. Geburtstag gewidmet.

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Mit Unterstützung der Deutschen Forschungsgemeinschaft und des Bundesministeriums für Atomkernenergie und Wasserwirtschaft.     

Die Wirkung von Erdalkalimetallionen auf die Hemmung des Kaliumverlustes fluoridvergifteter Erythrocyten durch Substrate und Stoffwechselgifte

Summary The inhibition of potassium loss of fluoride poisoned (40 mequiv/l. cell suspension) human red cells by metabolic poisons and substrates at varying Ca⁺⁺-concentrations has been studied. At low calcium concentrations, fluoride produces rapid potassium loss after a lag period, whereas sodium permeability is little affected. This specific action of fluoride on potassium permeability can be almost completely prevented by the addition of cyanide, monoiodoacetic acid or adenosine. At high calcium concentrations, exceeding 0.1 mequiv/l. cell suspension, fluoride destroys the permeability barrier for both sodium and potassium ions without a preceeding lag period, which eventually leads to colloid osmotic hemolysis. This effect cannot be prevented by adenosine, monoiodoacetic acid or cyanide. It is concluded that the permeability change at high calcium concentrations is independent of the action of fluoride on cellular metabolism and is due to the formation of a calcium-fluoride-complex with ligands in the outer cell surface. On the other hand, the specific effect on potassium permeability, occuring at low Ca⁺⁺-concentrations, is related to the inhibition of cellular metabolism by fluoride.

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Mit 3 Textabbildungen

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Herrn Professor Dr. Rudolf Mond zum 65. Geburtstag gewidmet.

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Mit Unterstützung der Deutschen Forschungsgemeinschaft und des Bundesministeriums für Atomkernenergie und Wasserwirtschaft.     

Deformierung und Zerstörung von Erythrocyten durch Noradrenalin in normalen und hämorrhagischen Hunden

Zusammenfassung Normalen Hunden wurde in Nembutal^®-Narkose Noradrenalin (NA) i. v. infundiert. Bei einer Infusionsrate von ca. 2 /min · kg veränderten die Erythrocyten ihre bikonkave Form. Sie wurden stechapfelörmig. 10 min nach Ende der Infusion waren alle Zellen wieder normal bikonkav. Die NA-Wirkung unterblieb bei hämorrhagischen Hunden (Blutdruck 40 mm Hg) und bei Steigerung der Infusionsrate auf 8 NA. Es wird diskutiert, ob NA die Fließeigenschaften der Erythrocyten durch Formveränderungen beeinflussen kann.

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Summary Anesthetized dogs received infusions of nor-adrenaline (0.5–8 /kg · min). During infusion the shape of the erythrocytes was observed in its own plasma in a coverglass preparation. 10 min after start of a 2 infusion up to 90% of the erythrocytes became more or less creanated. 10 min after stopping the infusion all erythrocytes showed normal biconcave shapes. Infusion of 8 /kg · min did not influence the shape. No changes were observed in dogs subjected to severe hemorrhage. During the hypotensive periode (40 mm Hg) neither the shape nor the concentration of free fatty acids responded to infusion of 0.5–2 NA/min · kg. It is discussed whether NA might disturb the microcirculation in capillaries by changing the flow properties of shape transformed erythrocytes.

     

Nachweis von Elastase in menschlichen Erythrocyten

Zusammenfassung Mit Hilfe einer photometrischen Methode (Kongorot-Elastin) lassen sich im Stroma und im stromafreien Hämolysat menschlicher Erythrocyten eine elastolytische Aktivität nachweisen und die gleichzeitige Anwesenheit eines Elastaseinhibitors vermuten.

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Summary Using a photometric method (kongored-elastin) we can prove in stroma and in supernatant of human erythrocytes an elastolytic activity. The simultaneous presence of an elastase inhibitor also may be probable.

     

Zum Sauerstoffverbrauch der Erythrocyten von Psoriatikern

Zusammenfassung Aufgrund des erhöhten Sauerstoffverbrauchs der Erythrocyten von Psoriatikern kann auf eine Steigerung des Pentosephosphatcyclus geschlossen werden. Dieser Befund weist gemeinsam mit anderen Untersuchungen auf das Vorliegen einer generalisierten Stoffwechselstörung bei der Psoriasis vulgaris hin.

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Summary The elevated oxygen-consumption of the erythrocytes of patients with psoriasis vulgaris can be interpreted as an increase of the pentose-cycle. These findings together with other results point to the existence of a generalized disorder in the psoriasis vulgaris.

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In dieser Arbeit sind Teile einer Inaugural-Dissertation mitverwertet.

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Herrn Prof. Dr. Dr. h. c. h. c. H. A. Gottron zum 80. Geburtstag verehrungsvoll gewidmet.