The Effect of Scalding on the Content of Kininogen and Kininase in Limb Lymph

Jacobsen, Sten Eirik W.; Waaler, B. A.

doi:10.1111/j.1476-5381.1966.tb01657.x

Cited by 9 publications

(5 citation statements)

References 11 publications

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“…The findings in lymph where the representation of the substrates is different relative to that in plasma (Jacobsen, 1966c;Jacobsen & Waaler, 1966) strongly support the concept of two substrates with different molecular dimensions. The available data thus indicate that in plasma there exist two different substrate entities, with different molecular dimensions, and that these two substrates are differently attacked by kinin-forming enzymes.…”

Section: Discussionsupporting

confidence: 55%

Some Data on Two Purified Kininogens From Human Plasma

Jacobsen

Kriz

1967

British Journal of Pharmacology and Chemotherapy

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Work on the purification of the substrate in plasma for kinin-forming enzymes, kininogen, has been carried out in several laboratories. Habermann (1963) thus purified kininogen from bovine serum and Suzuki, Mizushima, Sato & Iwanaga (1965) purified a kininogen from bovine plasma. Habermann (1965Habermann ( , 1966 also studied the structure of the kinin-yielding polypeptides of his purified bovine kininogen (peptic kinin-yielding polypeptides). Pierce & Webster (1966) isolated two kininogens from human plasma. These kininogens were shown to be glycoproteins with molecular weights of about 50,000, as is also the case with the bovine kininogen (Habermann, 1963). Jacobsen (1966b) found that two different substrates for plasma kinin-forming enzymes could be separated from plasmas of several mammals. These substrates had different molecular dimensions and they differed in their reaction patterns towards various kininforming enzymes. The present paper reports the results of further purification with some characterization of these two substrates. The substrate with the larger molecular dimension has been designated substrate 1, and the other and smaller one has been designated substrate 2. The methods employed have been DEAE anion exchange chromatography, gel filtration, disc gel electrophoresis and sucrose gradient ultracentrifugation. METHODS Plasma. Citrated human plasma was obtained by using siliconized equipment throughout, as described by Jacobsen (1966b).Initial purification procedures for kininogens (anion exchange chromatography and gel filtration). The separation procedures previously described (Jacobsen, 1966a(Jacobsen, , 1966b have been modified and expanded, so as to include disc gel electrophoresis (see below). The initial procedure was a separation of fractions containing the two kininogens by chromatography of human citrated plasma on a DEAE anion exchanger. Each of the substrates was subsequently exposed to further purification procedures (see below and Table 1). For initial chromatography, plasma (25 ml.) was mixed with 0.05 M Tris-HCl buffer (pH 8) containing 0.25 M sodium chloride (12.5 ml.) and the mixture applied to a DEAE-Sephadex A 50 (A. B. Pharmacia, Uppsala, Sweden) column (18 x 1.8 cm) equilibrated with 0.05 M Tris-HCl buffer (pH 8) containing 0.2 M sodium chloride. Substrate 1 is retained on the column while substrate 2 is eluted with the equilibrating buffer. Substrate 1 could then be eluted with a 0.05 M Tris-HCl buffer (pH 8) containing 0.35 M sodium chloride. The effluent containing substrate 2 was treated with " massive contact exposure " in order to eliminate the plasma kallikrein activity (Jacobsen, 1966b), and then applied to a DEAE-Sephadex A 50 column (10 x 1 cm) equilibrated with 0.05 M Tris-HCI buffer (pH 8) containing 0.1 M sodium chloride. Substrate 2 was eluted from this column with a buffer containing 0.25 M sodium chloride. Buffer concentrations in the samples were always adjusted to that of the buffer used for equilibration in the following purification procedure. S. JACOB...

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Section: Discussionsupporting

confidence: 55%

Some Data on Two Purified Kininogens From Human Plasma

Jacobsen

Kriz

1967

British Journal of Pharmacology and Chemotherapy

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“…It has been found that the protein content of the perfusates parallels that of bradykinin and it thus appears that during the heating process protein migrates to the tissue spaces first and this is then followed by water to give the oedmea reaction. It may be that the pH value of the interstitial fluid in the paws subjected to thermal injury reaches a sufficiently low value to activate the kinin-releasing enzyme, for this enzyme is easily activated in rat plasma (Jacobsen & Waaler, 1966).…”

Section: Discussionmentioning

confidence: 99%

Bradykinin and Oedema Formation in Heated Paws of Rats

Starr

West

1967

British Journal of Pharmacology and Chemotherapy

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Histamine was considered to be the chief mediator in inflammatory reactions until it was shown that specific antihistamine drugs did not always inhibit-these reactions. The idea of the polypeptide nature of some of the unidentified permeability factors in inflammatory exudates was introduced by Menkin (1936). Armstrong, Jepson, Keele & Stewart (1957) noted the similarity of the actions of these factors with those of bradykinin, a nonapeptide. This compound or others related to it have since been found in many oedema fluids and were detected in the perfusates of rat paws after the application of heat (Rocha e Silva & Antonio, 1960).It was considered important to investigate in more detail the extent to which kinins participate in this so-called thermic oedema by following changes in four-kinin parameters in the perfusates. As rats which are resistant to the anaphylactoid reaction produced by dextran (the so-called non-reactors) also showed greater resistance-to thermic oedema than did the reactors giving the anaphylactoid reaction (Gecse, Karady, Starr & West, 1965), a comparison of the activity of the kinin systems in the perfusates collected from the heated paws of both types of rat was also made. In addition, a study of the effects of drugs on both the development of thermic oedema and the actions of bradykinin has been included. METHODSAdult Wistar albino rats were obtained from Fison's Ltd. (Holmes Chapel) and from the Agricultural Research Council's Field Station at Compton. They were tested for their reactivity to dextran (240 mg/kg, intraperitoneally) on three occasions, oedema developing in the extremities over a 2-hr period only in reactor rats (Harris & West, 1961). Both reactor and non-reactor rats were anaesthetized with pentobarbitone sodium (45 mg/kg, intraperitoneally) and their hind paws were perfused according to the method of Rocha e Silva & Antonio (1960). Before collecting the perfusates, the paw space was washed free of blood. The flow of perfusing fluid (Tyrode solution) was regulated to 5-10 drops/min, and then the paw was immersed in a water-bath maintained at the desired temperature for 30 min. Perfusates were collected for three consecutive periods of 10 min during the heating and for further periods after the heating. Samples were also collected from unheated paws to serve as controls.In other experiments, paws were heated without perfusion after the rats had been anaesthetized and given azovan blue dye intravenously (18 mg/kg, Bonaccorsi & West, 1963). The paws were then examined for the presence of oedema using the plethysmographic method of Buttle, D'Arcy,

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“…The object of these estimations was to ensure that during the assay of lymph kallikrein activity, more than sufficient kininogen was present without the addition of substrate prepared from other animals. There was no doubt that there was considerably more kininogen present than that utilized during the assay of kallikrein showing that the increase in the activity of acid kallikrein was not due to the passage of substrate into the lymph following an increase in vascular permeability, as suggested by Jacobsen & Waaler (1966). Further, in the experiments in which exogenous kininogen was added (see Table 2), the concentration of kinins formed was not higher than those in Tables 3 and 4 in which no additions of kininogen were made.…”

Section: Discussionmentioning

confidence: 94%

“…Some of the difficulties were pointed out by Jacobsen & Waaler (1966) who reported that the increase of kinin forming potential in lymph after injury was due to an increase in kininogen concentration resulting from an increase in vascular permeability.…”

Section: Introductionmentioning

confidence: 99%

Effect of thermal injury on the kinin system in rabbit hind limb lymph

Lewis¹,

Wawretschek²

1971

British J Pharmacology

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Summary The kinin‐forming activity of hind limb lymph and of plasma has been examined in rabbits before and after thermal injury. Neither plasma nor lymph contained much active kallikrein activity but the enzyme was evident in samples treated with glass or with acid. There was little or no increase in the activity of enzyme activated by glass after thermal injury, but an increase in the activity of enzyme activated by acid regularly occurred. There were two increases in the activity of enzyme activated by acid—one about 2 h and the other 4–6 h after thermal injury. They corresponded to increases in vascular permeability as indicated by increases in the concentration of lymph protein. There was considerably more kininogen in the lymph and plasma than was used in the assays of kallikrein activity, showing that the increased kinin‐forming activity in lymph was not the result of the passage of kininogen from the plasma. The increase in activity in lymph was not usually accompanied by a similar increase in the plasma. However, an increase in the activity of enzyme activated by acid sometimes occurred in the plasma simply as a result of prolonged anaesthesia. It is suggested that whereas the enzyme activated by glass is a measure of prekallikrein, the acid activatable enzyme appears as a result of the dissociation of a kallikrein‐inhibitor complex. An increase in the concentration of this complex is therefore an indication of the preceding activation of kallikrein.

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The Effect of Scalding on the Content of Kininogen and Kininase in Limb Lymph

Cited by 9 publications

References 11 publications

Some Data on Two Purified Kininogens From Human Plasma

Some Data on Two Purified Kininogens From Human Plasma

Bradykinin and Oedema Formation in Heated Paws of Rats

Effect of thermal injury on the kinin system in rabbit hind limb lymph

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