SVIBOR - Project code: 3-03-312

MINISTRY OF SCIENCE AND TECHNOLOGY

Strossmayerov trg 4, HR - 10000 ZAGREB
tel.: +385 1 459 44 44, fax: +385 1 459 44 69
E-mail: ured@znanost.hr

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Project code: 3-03-312


IMMUNOLOGICAL ACTIVITY AND APPLICATION PREPARING OBTAINED FROM EARTHWARM (fam. Lumbricidae)


Main researcher: HRŽENJAK, TEREZIJA (16733)



Assistants
Type of research: applied
Duration from: 01/01/91. to 01/01/96.

Papers on project (total): 20
Papers on project quoted in Current Contents: 5
Institution name: Veterinarski fakultet, Zagreb (53)
Department/Institute: Department of biology
Address: Heinzelova 55
City: 10000 - Zagreb, Croatia
Communication
Phone: 385 (01) 2390-142
Fax: 385 (01) 214-697

Summary: Earthworms (Annelida, Oligochaeta) are one of the first organisms in the evolution of the any three that possess immunological recognition and memory. If the nervous system is not removed from the regenerative area they can rebuild an amputated part of their body. Their body fluid, i.e. coelomic fluid possesses agglutinins, opsonins, lysisns, mitogens and machrophage inhibition factor. Their position in evolution shows that genetic basis of macromolecules which show those characheristica did not change very much. The aim of the research was to isolate biologically active macromolecules, to determine their genetic basis and to find appropriate practical application. From tissue homogenate Eisenia foetida (fam. Lumbricidae) glycolipiprotein extract G-90 was separated. It shows biological activities: mitogenic, cytotoxic, bacteriostatic, tumorostatic selective agglutinative and haemolytic activites. In this research those macromolecules would be isolated from G-90 and tested "in vitro" and "in vivo" to determine their function and specificity. Macromolecules will be biochemically and genetically examined. We expect that G-90 contains enzymes, growth factors and lectines. Their isolation would be contribution to study of transfer of signals from plasma memrane to nucleus, it would show evolution way of some macromolecules and to be a basis for drugs with tumorostatic effect.

Keywords: earthworms, glycolipoproteins, lectins, tumorostatics, bacteriostatics, immunomodulators, growth factors, enzymes

Research goals: Aimes: The biologically active macromolecules carriers of activities such as agglutination, opsonisation, regeneration and mitogenicity would be isolated from wet tissue of adult earthworms. The biologica and biochemicall characteristions, as well as applications of them will be investigated (immunomodulators, tumorostatics). Results a biologically active glycolipoprotein extract from a whole earthworm tissue homogenate was isolated and named G-90. G-90 increases (3H) tymidine incorporation into melanoma B16, fibrosarcoma CMC1, HeLa cella and health murine lymphocytes. Stimulation or inhibition depends on of concentrations of G-90. It alters murine cell growth rate "in vitro" in serum in a dose dependent manner and slows murine tumor growth "in vivo". G-90 was separated in seven fractions by gel filtrations. It has been shown by RIA and Western blotting that fractions is cross-reactive with porcine anti-insulin antibodies. We consider, a new "growth factor" exists in the supstance. It could be a growth factor from the family of "insuline like growth factors". The supstance is prepared to determination genetic code. We can not continue this investigations because of insuficient finantial support. From G-90 two enzymes were isolated. Thay have got a serin-proteolitic activity, molecular weight of 23 and 28 kDa. Investigations are part of master thesys of "the young investigator". G-90 forms precipitation arcs in gel with different animal and human sera. We fortify, the interactions yeald with sera IgA.


COOPERATION - PROJECTS


  1. Name of project: 3-03-464 IMUNOGLOBULINI A U DIJAGNOSTICI HUMANE I VETERINARSKE MEDICINE
    Name of institution: VETERINARSKI FAKULTET, SVEUČILIŠTE U ZAGREBU
    City: 10000 - Zagreb, Croatia


COOPERATION - INSTITUTIONS


  1. Name of institution: Institut Ruđer Bošković, Zagreb, Hrvatska
    Type of institution: University/Faculty
    City: 10000 - Zagreb, Croatia

  2. Name of institution: Pliva, farmacija, kemijska kozmetika i prehrambena industrija
    Type of institution: State institute
    City: 10000 - Zagreb, Croatia

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Last update: 10/11/95
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