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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 265 (1970), S. 442-454 
    ISSN: 1432-1912
    Keywords: Haemolysis ; Phospholipase A ; Direct Lytic Factor ; Polypeptides ; Toxins ; HÄmolyse ; Phospholipase A ; Direkt lytischer Faktor ; Polypeptide ; Toxine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The haemolytic action on washed guinea-pig red cells of the following substances has been studied: the direct lytic factor (DLF) of cobra venom, melittin and an apamin-containing fraction of bee venom, anaphylatoxin (AT), angiotensin, vasopressin, saponin, p-chloro-mercuribenzoate (p-CMB) and N-ethylmaleimide (NEM). Further the synergism of these substances with phospholipase A in causing haemolysis has been investigated. In regard to the lytic effects, the substances studied can be classified as follows. 1. Substances which react with SH-groups, either by means of -S-S- bonds (DLF, apamin-fraction, AT, vasopressin) or by other structures (p-CMB, NEM) produce weak or no direct haemolysis, but strongly potentiate haemolysis caused by phospholipase A. Their effect is increased by Ca++, inhibited by EDTA, and strongly dependent on temperature (as far as has been investigated). 2. Angiotensin, a peptide without disulfide groups, is not haemolytic, neither directly nor in combination with phospholipase A. Saponin, which does not react with SH-groups, also does not show potentiated haemolysis with phospholipase A in spite of being haemolytic itself. 3. Melittin, though not containing disulfide structures, does produce potentiated haemolysis with phospholipase A, even at concentrations which are not lytic when acting alone. It is concluded that more than one mechanism of potentiating phospholipase A haemolysis exists. One possibility is the reaction of potentiating agents with SH-groups of membrane constituents (enzymes?) of the red cells. This mechanism applies to p-CMB, NEM and to disulfide-containing peptides. It is independent of detergent effects. Another mechanism may be membrane changes due to a lowering of surface tension such as that produced by melittin. It seems doubtful, however, whether this is the only molecular property responsible for the potentiation, as the detergent saponin does not have such an effect. Possibly melittin, in addition to having detergent effects interferes with the same membrane properties which are altered by the SH-reactants.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 275 (1972), S. 203-211 
    ISSN: 1432-1912
    Keywords: Glutathione Reductase ; Glutathione ; Haemolysis ; Direct Lytic Factor ; Disulphides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The activity of glutathione reductase (GR) was measured in haemolysates and red cell membranes of various species. The enzyme levels were compared with the susceptibility of the respective cells to the direct lytic factor (DLF) of cobra venom. A positive correlation was found in so far as cells having high (low) GR activity were highly (little) sensitive to DLF. Further results make it likely that GR is implicated in DLF-induced lysis: 1. By enzymatic assay an interaction of membrane bound GR and DLF was found, as evident from consumption of the coenzyme NADPH. 2. Glutathione in the reduced state (GSH) as well as in the oxidized form (GSSG) inhibited haemolysis by DLF in a dose-dependent manner. A chemical interaction between DLF and glutathione was excluded. 3. NADPH showed a dual effect: it accelerated DLF-induced haemolysis at low concentrations, whereas at high concentrations inhibition was evident.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 282 (1974), S. 255-260 
    ISSN: 1432-1912
    Keywords: Direct Lytic Factor ; Cobra Venom ; Phospholipase A ; Red Cells ; Haemolysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The effect of increasing the (colloid-)osmotic pressure in the extracellular medium on haemolysis by the direct lytic factor of cobra venom (DLF) and phospholipase A has been investigated. For comparison, N-ethyl-maleimide (NEM) and p-chloromercuribenzoate (p-CMB) were used. Dextran and sucrose abolished the haemolytic effect of NEM and p-CMB but reduced only slightly (dextran) or not (sucrose) the weak lytic activity of DLF. Haemolysis by phospholipase A in the presence of DLF, NEM or p-CMB was not significantly inhibited. Hypertonic NaCl solution considerably retarded the onset of haemolysis by DLF plus phospholipase A. The mean corpuscular volume of guinea-pig red cells increased slightly but definitely during incubation with DLF. It is concluded that the haemolytic effect of DLF has non-osmotic as well as osmotic components, and that phospholipase A causes non-osmotic haemolysis. The retardation of haemolysis by hypertonic NaCl probably indicates specific inhibition of bee venom phospholipase A2, not protection of the erythrocytes from osmotic stress.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 280 (1973), S. 201-207 
    ISSN: 1432-1912
    Keywords: Direct Lytic Factor ; Cobra Venom ; Red Cells ; Membranes ; Haemolysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The binding of direct lytic factor (DLF) from cobra venom (Naja naja) to intact guinea-pig red cells and to guinea-pig ghosts was estimated quantitatively by bioassay of DLF in the supernatant. 1. DLF was not bound to intact red cells in considerable amounts, during 320 min incubation. 2. The degree of binding to ghosts was much larger than that in suspensions of intact red cells. Binding to ghosts increased with time. 3. Whereas the binding of DLF to ghosts was not much influenced by varying the incubation temperature, its haemolytic activity was completely absent at temperatures below 15°C. By an immunofluorescence technique binding of DLF to erythrocytes was studied morphologically: 1. DLF was only bound to red cell ghosts (guinea pig and rat), but not to intact red cells. This binding was not temperature dependent. 2. Pretreatment of ghosts with SH-reagents such as NEM or PCMB did not prevent binding of DLF. 3. Ghosts prepared by different methods (hypotonic shock, freezing and thawing, ultrasonication, and resealing) were all able to bind DLF to their surface. It is concluded that the binding of small amounts of DLF to intact red cells, observed by bioassay, was due to the presence of a small fraction of lysed cells, and that the binding to ghosts is not related to the lytic effect of DLF but secondary to lysis.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 274 (1972), S. 81-90 
    ISSN: 1432-1912
    Keywords: Direct Lytic Factor ; Phospholipase A ; Red Cells ; Ion Permeability ; Haemolysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The effects of the direct lytic factor (DLF) of cobra venom, bee venom phospholipase A and of combinations of these two lysins on guinea-pig red cells have been studied. DLF caused an increased permeability to Na ions, swelling of the cells and moderate haemolysis. No prelytic loss of potassium was seen. Phospholipase A produced loss of potassium, considerable gain of sodium, cell swelling, but no remarkable lysis. Combinations of the two venom constituents, more strongly haemolytic, mimicked the effect of the predominant component of the mixture. Thus prelytic loss of potassium was observed when higher concentrations of phospholipase were combined with low concentrations of DLF, but was absent when DLF predominated. The rather low critical volume and spherocytosis of haemolysing red cells exposed to DLF, and the effect of DLF on osmotic stability suggest that DLF has other effects on the cell membrane in addition to those of an osmotic haemolysin.
    Type of Medium: Electronic Resource
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