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  • 1990-1994  (3)
  • Biochemistry and Biotechnology  (2)
  • Fibrinogenolysis  (1)
  • Analytical Chemistry and Spectroscopy
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Years
Year
  • 1
    ISSN: 1433-2981
    Keywords: Hementin ; Antihaemostatic factors ; Salivary gland complex ; Giant Amazon leech ; Fibrinogen ; Fibrinogenolysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Biological observations were made on the antihaemostatic activity in the saliva of the giant Amazon leech Haementeria ghilianii. Particular emphasis was placed on determining where the fibrino(geno)lytic enzyme hementin is produced and secreted in the salivary gland complex. Hungry third-fed Amazon leeches produce about 650 units of hementin, with by far the majority of the activity (75%) in the two posterior glands. The two anterior glands contain much less hementin (12%), and a small amount (12%) resides in the proboscis itself, presumably in the salivary gland ductules. In the anterior gland, hementin appears to be produced by only certain cells. The secretion of hementin is confined to the lumen of the proboscis. Secretions in the proboscis lumen are rich in antihaemostatic activity, as evidenced by fibrinogenolysis (hementin), prolongation of prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin clotting time (TCT) and Atroxin clotting time, and inhibition of collagen-induced platelet aggregation. In contrast, no anti-haemostatic, including hementin, activity was detectable at the tip. Suprisingly, the wound from the bite by the Amazon leech is not associated with prolonged bleeding. This is in marked contrast to the wound by the European medicinal leech Hirudo medicinalis which bleeds for an average of ten hours. The absence of bleeding is compatible with the interpretation, based on the above findings, that during feeding the Amazon leech does not inject antihaemostatic factors into the host, or at least not in the vicinity of the wound.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0952-3499
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A secreted form of phospholipase A2 (PLA2) is thought to play an important role in inflammatory diseases. To characterize this enzyme the cDNA encoding a low molecular weight PLA2 was cloned from a human placental cDNA library. The cDNA encoding the human PLA2 was subcloned into an expression vector and subsequently transfected into Chinese hamster ovary (CHO) cells. A stable CHO cell clone, secreting ca I mg/L of recombinant PLA2 into the medium, was scaled up in culture to 180L. The recombinant enzyme was purified from the cell supernatant to apparent homogeneity by a novel procedure combining adsorption to poly(vinylidene difluoride) membranes, ion exchange chromatography and size exclusion chromatography. The final recovery of PLA2 activity was 58%. A direct comparison between the purified recombinant human PLA2 and PLA2 purified from human synovial fluid, including molecular weight, antigenicity, ionic dependence, substrate specificity and sensitivity to know PLA2 inhibitors, indicated that the two enzymes exhibit identical biochemical properties. These results show that the recombinant PLA2 can be efficiently expressed and purified in sufficient quantities to characterize the enzyme active site, to aid in the rational development of PLA2 inhibitors as potential anti-inflammatory drugs, and to investigate further the role of PLA2 in inflammatory disease.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Chichester : Wiley-Blackwell
    Journal of Chemical Technology AND Biotechnology 59 (1994), S. 115-115 
    ISSN: 0268-2575
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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