WO1988000056A1 - Composition et procede de castration et de sterilisation immunologiques - Google Patents
Composition et procede de castration et de sterilisation immunologiques Download PDFInfo
- Publication number
- WO1988000056A1 WO1988000056A1 PCT/AU1987/000199 AU8700199W WO8800056A1 WO 1988000056 A1 WO1988000056 A1 WO 1988000056A1 AU 8700199 W AU8700199 W AU 8700199W WO 8800056 A1 WO8800056 A1 WO 8800056A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- lhrh
- composition
- animals
- rams
- protein
- Prior art date
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0005—Vertebrate antigens
- A61K39/0006—Contraceptive vaccins; Vaccines against sex hormones
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/23—Luteinising hormone-releasing hormone [LHRH]; Related peptides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
- A61K2039/6031—Proteins
- A61K2039/6037—Bacterial toxins, e.g. diphteria toxoid [DT], tetanus toxoid [TT]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
- A61K2039/6031—Proteins
- A61K2039/6068—Other bacterial proteins, e.g. OMP
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
- A61K2039/6031—Proteins
- A61K2039/6081—Albumin; Keyhole limpet haemocyanin [KLH]
Definitions
- the present invention relates to the suppression of social and sexual behaviour of male domestic animals and the suppression of ovulation and oestrous cyclicity of female domestic animals, and more specifically to a composition and method for immunological castration and spaying.
- LHRH luteinizing hormone releasing hormone
- LHRH is released from the hypothalamus into the pituitary portal vessels. It has been suggested that LHRH binds to the gonadotrophs of the anterior pituitary to stimulate secretion and release of luteinizing hormone (LH). LH acts on the Leydig cells in the testis to increase the production of testosterone. Active immunization against LHRH disrupts the communication between the hypothalamus and pituitary. It is thought that the site at which antibodies to LHRH might inhibit reproductive function is in the hypophyseal portal blood vessels (Schanbacher, 1984).
- LHRH is a small peptide of 9 amino acids. In the field of immunology, it is well established that small immunogens elicit a poor immune response when injected into a host. Consequently, in each of the above studies, LHRH has been chemically conjugated to one particular carrier protein, for example, Human Serum Albumin.
- the present applicants have surprisingly found that a synergistic interaction occurs when animals are immunised with a combination of two or more different carriers coupled to LHRH or analogues of LHRH.
- Combinations of LHRH or analogues of LHRH conjugated with carriers elicit an effective immune reponse (measured in terms of antibody titre) in a greater proportion of animals than if LHRH or analogues of LHRH conjugated to a single carrier are administered.
- compositions for the production in animals of antibodies specific for LHRH or analogues of LHRH comprising two or more different carriers individually coupled to LHRH or analogues of LHRH.
- Analogues of LHRH are peptides where additions, deletions, modifications or alterations have been made to the natural sequence of LHRH. Alterations to the natural sequence include the addition of one or more amino acids to the amino or carboxy terminus of LHRH.
- Domestic animal species falling within the scope of the invention include, cattle, sheep, goats, cats, guinea pigs, pigs, dogs, reindeer, horses and primates including women.
- Carriers of any type which are useful immunogens may be utilised.
- the carriers are proteins and more preferably the proteins are soluble in aqueous solution although precipitated protein may be used.
- DT Diptheria Toxoid
- HSA Human Serum Albumin
- P.A. staphyloccus protein A
- KLH Keyhole Limpet Haemocyannin
- TT Tetanus Toxoid,
- the carriers are proteins such as, HSA and P.A. or KLH and TT, each of these proteins being conjugated to LHRH.
- two or more carriers where each carrier coupled to LHRH and separately administered toan animal elicits a generally good titre response.
- compositions where one of the carriers is Diptheria toxoid (DT).
- Other carriers that may be used in the present invention include lipopolysaccharides, polysaccharides, glycopeptides, muramyl peptide analogues, peptidoclycans [from bacteria cell walls], liposomes, lecithin like materials, and bacterial proteins for example purified protein derivatives from tuberculin.
- the composition contains an adjuvant.
- adjuvants which may be used are aluminium hydroxide, Freunds incomplete adjuvant, Freunds complete adjuvant, DEAE dextran, levamisole, PCG and polyA, polyC or polyU.
- a mineral oil composition that includes bacterial cell wall material such as peptidoglycans, or a synthetic derivative of such a cell wall material. Such a synthetic material is known as muramyl dipeptide.
- composition may be optionally buffered to physiological pH using buffers such as TRIS-HCl, Hepes, pipes or other appropriate buffers.
- buffers such as TRIS-HCl, Hepes, pipes or other appropriate buffers.
- a method for the suppression of male social and sexual behaviour of domestic animals comprising the administration of a composition to a domestic animal said composition comprising two or more different carriers individually conjugated to LHRH or analogues of LHRH.
- Administration may be parenterally, for example, subcutaneous, and/or intramuscular or intravenous injection, orally or by absorption through the skin or by a mini-pump either implanted in the animal or attached to the outside of the animal.
- bull calves are preferably immunised with the composition at between 8 to 40 weeks of age.
- ram lambs are preferably immunised at 8 to 24 weeks.
- Booster injections are preferably given eight weeks apart and preferably 1 to 3 booster injections are given.
- the administration of the substance e.g. LHRH
- the present invention has particular utility in the immunological castration and spaying of domestic pets or companion animals, particularly cats and dogs. Conventional surgical treatments performed by veterinarians may be expensive, and often traumatic for the animal involved and its owner.
- An increase in the proportion of animals responding; may also occur where other substances that elicit an immune response, other than LHRH or analogues of LHRH, are conjugated to at least two different carriers and combinations of these conjugates are subsequently administered to animals.
- peptides and proteins such as adrenocorticotrophic hormone, substance P, human chorionic gonadotrophin, somatostatin, epidermal and insulin-like growth factors; steroids including androstenedione (fecundin), testosterone and other substances such as melatonin and the prostaglandins.
- LHRH Luteinizing hormone releasing hormone is a specific substance involved in stimulating luteinizing hormone and follicle stimulating hormone from the adenohypophysis.
- Carrier Any substance, especially protein which, when coupled to small substances, can elicit an immune response to animals.
- Antibody titre The dilution of antisera which binds 50% of 0.15 nCi 131 I-labelled LHRH for 16 h at 4°C. Precipitation of the gamma globulins was achieved using polyethylene glycol at a final concentration of 14.5%.
- HSA Human serum albumin
- CP Corynebacterium parvum.
- P.A. Protein A from Staphylococcus aureus.
- Carb 1-ethyl-3 (3-dimethyl aminopropyl) carbodiimide.
- Glut Gluteraldehyde.
- Figures 1, 2 show Antibody Titre (Log Scale) plotted against various treatment groups for ram lambs and bull calves respectively.
- Figures 3 and 4 show mean testicular volume of ram lambs given Corynebacterium Parvum (CP), Keyhole Limpet Haemocyanim (KLH), Diphtheria Toxoid (DT), Testanus Toxoid (TT) and a combination of these carriers coupled to LHRH; CP, KLH, DT in Figure 4 and KLH, TT and DT in Figure 3.
- CP Corynebacterium Parvum
- KLH Keyhole Limpet Haemocyanim
- DT Diphtheria Toxoid
- TT Testanus Toxoid
- LHRH (240 mg) and tetanus toxoid (35.5 ml, 248 mg) was dissolved with 20 ml distilled water and the pH of the mixture adjusted to 6.0 with 0.1 N NaOH.
- a freshly prepared solution of 0.4 mg 1-ethyl-3 (3-dimethyl amino propyl) carbodimide hydrochloride (ECDI) in 2 ml distilled water has added dropwise and the reaction allowed to proceed at room temperature (22oC) overnight.
- the solution was dialysed against 3 changes of saline (5 litres). To estimate the number of moles of LHRH bound to tetanus toxoid 3nCi of 131 I labelled LHRH was added into the reaction mixture.
- the solution was dialysed against 3 changes of saline (5 litres).
- a flock of 42 Merino ram lambs were used in the study.
- the rams were selected by liveweight into 7 groups of 6 rams. (However during the course of the experiment some rams were lost; deaths were attributed to pneumonia). Treatments are shown below:
- Each individual immunogen solution was emulsified with equal volumes of Freund's complete adjuvant.
- the ram lambs were then given 2 ml of the mixture in 4 separate subcutaneous sites in the back leg adjacent to the groin.
- the combined immunogens were injected with 8 ml of the emulsion.
- Booster injections were made using identical quantities of freund's incomplete adjuvant.
- a herd of 35 bull calves of predominently Friesian breed were used for the experiment. The calves were raised on skim milk and weaned onto grass at 10 weeks of age. The bulls were divided into 5 groups of 7 animals using liveweight as the basis of selection (during the course of the experiment three bulls were lost due to pneumonia). Treatments are shown below.
- composition, dose and immunization schedule for the experiment is outlined in Table 2.
- the immunogen solution was emulsified in an equal volume of Freund's complete adjuvant.
- Freund's complete adjuvant For all individual treatment groups a total of 2 ml of the emulsion solution was injected into 4-5 sites.
- the bull calves were injected into 4-5 sites with 8 ml of the emulsion solution. Injections were given subcutaneously near the front shoulder and the brisket of the calves.
- the immunogens were emulsified using Freund's incomplete adjuvant. Blood samples were collected into heparinized containers at two weeks after the second booster injection. The antibody titres for this experiment are given in Table 4.
- the behavioural data presented in Table 5 show that control animals spent more time involved in social and sexual behaviour and in social and sexual investigation than animals in the castrate, combination (LHRH individually coupled to KLH, TT and PA), and tetanus toxoid (TT coupled to LHRH) treatments.
- LHRH individually coupled to KLH, TT and PA
- TT tetanus toxoid
- a flock of 80 Merino X Corredale rams at 5 months of age were used in this experiment.
- the rams (16 weeks of age) were allocated to 8 groups of 10 rams. One group were surgically castrated at 16 weeks and another group were left as entire animals. The remaining 60 rams were treated with a number of LHRH antigens.
- Various LHRH - carrier protein conjugates were prepared as described previously. The antigens used were LHRH coupled separately to diphtheria toxoid, tetanus toxoid, keyhole limpet haemocyanin and corynebacterium parvum. Two other groups were injected with two different combinations of the individual LHRH complexes.
- Liveweights and testicular size (as determined using a series of calibrated beads) was measured every 4 weeks.
- Antibody titre to LHRH were measured by both radioimmunoassay (RIA) and ELISA techniques.
- Plasma samples were serially diluted with phosphosaline buffer (0.05M, pH 7.6). The diluted samples (100 ul) were added to 100 ul bovine gamma globulin (3% 9w/v) and
- Liveweights of all rams are shown in Table 7. There were no significant differences among groups although the mean liveweight values for the immunised rams were higher than the castrated males but lower than the control rams.
- the mean testicular volumes for each treatment are shown in Table 8 and Figures 3 and 4. There was considerable variation with each individual antigen treatment. The largest decrease for individual carriers was observed in the rams immunised against LHRH coupled to diptheria toxoid. The next best carriers were tetanus toxoid follwed by keyhole limpet haemocyanin and corynebacterium parvum. The two different combinations of 3 separate antigens gave lower testes volumes than each of the individual antigens.
- the proportion of rams in each group that were sexually active at 2 months after the booster injection are shown in Table 10. Ten out of a total of 10 rams in the control group were showed sexual activity. The best single antigen was diphtheria toxoid with 1 of 9 rams sexually active. The order of effectiveness was diphtheria toxoid, tetanus toxoid, keyhole limpet haemocyanin and corynebacterium parvum. The two groups with the combined antigens were not sexually active. For those rams given a combination of TT, DT and KLH coupled to LHRH there were no rams active out of a total of 10 rams. Similar results were obtained using a different combination of carriers, DT CP and KLH where again there were no rams active out of a total of 8 rams (two rams died during the trial). None of the castrated rams were sexually active.
- T974 Blockage of the preovulatory surge of LH and FSH and ovulation by anti-LHRH serum in rats. Endocrinology 95:323-325.
- Clarke et al. (1978) Active immunization of ewes against luteinizing hormone releasing hormone, and its effect in ovulation and gonadotrophin, prolactin and ovarian steroid secretion.
- Robertson et al. (1984) Immunological castration of young bulls for beef production. In “Manipulation of growth in farm animals” (Eds. Roche, J.F. and D'Callaghan, D.) pp. 137-145.
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Epidemiology (AREA)
- Endocrinology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Immunology (AREA)
- Animal Behavior & Ethology (AREA)
- Virology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
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Abstract
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FI880954A FI880954A (fi) | 1986-07-03 | 1988-03-02 | Komposition och foerfarande foer immunologisk kastrering. |
NO880918A NO880918L (no) | 1986-07-03 | 1988-03-02 | Preparat og metode for immunologisk kastrering og sterilisering. |
DK114588A DK114588A (da) | 1986-07-03 | 1988-03-03 | Sammensaetning og fremgangsmaade til immunologisk kastration og udboedning |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AUPH672186 | 1986-07-03 | ||
AUPH6721 | 1986-07-03 | ||
AUPH9205 | 1986-11-27 | ||
AUPH920586 | 1986-11-27 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1988000056A1 true WO1988000056A1 (fr) | 1988-01-14 |
Family
ID=25643122
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/AU1987/000199 WO1988000056A1 (fr) | 1986-07-03 | 1987-07-03 | Composition et procede de castration et de sterilisation immunologiques |
Country Status (6)
Country | Link |
---|---|
EP (1) | EP0274496A4 (fr) |
JP (1) | JPH01500900A (fr) |
DK (1) | DK114588A (fr) |
FI (1) | FI880954A (fr) |
NZ (1) | NZ220932A (fr) |
WO (1) | WO1988000056A1 (fr) |
Cited By (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE4003944A1 (de) * | 1990-02-14 | 1991-08-22 | Gerhard Dr Prinzhaus | Synthetisches polypeptid: lh - rh -- klh (konjugat aus luteinisierendem hormon-releasing hormon und dem keyhole limpet haemocyanin |
EP0501882A2 (fr) * | 1991-03-01 | 1992-09-02 | Rhone Merieux | Procédé d'immunoneutralisation anti-LHRM des animaux domestiques mâles non castrés et peptide pour cela |
EP0578293A1 (fr) * | 1992-06-18 | 1994-01-12 | Akzo Nobel N.V. | Protéine frangée (fimbrilline-P) de E.Coli comme porteur immunogène contre GnRH |
EP0596959A1 (fr) * | 1991-07-26 | 1994-05-18 | Commonwealth Scientific And Industrial Research Organisation | Systeme d'administration de vaccin peptidique produisant son propre adjuvant et production du vaccin |
US5403586A (en) * | 1989-08-25 | 1995-04-04 | Biotechnology Australia Ptl Ltd. | LHRH-TraTp fusion proteins |
US5688506A (en) * | 1994-01-27 | 1997-11-18 | Aphton Corp. | Immunogens against gonadotropin releasing hormone |
WO1998027111A1 (fr) * | 1996-12-17 | 1998-06-25 | Centro De Ingenieria Genetica Y Biotecnologia. (Cigb) | Vaccin pour l'immuno-castration reversible des mammiferes |
WO1998051339A1 (fr) * | 1997-05-14 | 1998-11-19 | Pasteur Merieux Serums & Vaccins | Composition vaccinale multivalente a porteur mixte |
EP1007084A1 (fr) * | 1997-07-09 | 2000-06-14 | Csl Limited | Composition immunogene de lh-rh et procedes concernant cette composition |
EP3466441A3 (fr) * | 2012-03-09 | 2019-06-19 | Nord University | Procédés immunologiques d'inhibition de la maturation gonade |
WO2020051656A1 (fr) * | 2018-09-12 | 2020-03-19 | Ouro Fino Saúde Animal Ltda | Polypeptide chimérique antigénique, construction génétique et composition antigénique pour l'immunocastration de mammifères non humains |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA1239346A (fr) * | 1985-06-04 | 1988-07-19 | Gursaran P. Talwar | Vaccin anticonceptionnel |
-
1987
- 1987-07-02 NZ NZ22093287A patent/NZ220932A/xx unknown
- 1987-07-03 EP EP19870904375 patent/EP0274496A4/fr not_active Withdrawn
- 1987-07-03 WO PCT/AU1987/000199 patent/WO1988000056A1/fr not_active Application Discontinuation
- 1987-07-03 JP JP50402287A patent/JPH01500900A/ja active Pending
-
1988
- 1988-03-02 FI FI880954A patent/FI880954A/fi not_active IP Right Cessation
- 1988-03-03 DK DK114588A patent/DK114588A/da not_active Application Discontinuation
Non-Patent Citations (2)
Title |
---|
CHEMICAL ABSTRACTS, Volume 98, No. 17, issued 1983, April 25 (Columbus, Ohio, U.S.A.), DONNA L. VOGEL et al, 'Sertoli Cell Maturation is Impaired by Neonatal Passive Immunization with Antiserum to LHRH', Abstract No. 137896q. * |
See also references of EP0274496A4 * |
Cited By (25)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6019983A (en) * | 1982-06-19 | 2000-02-01 | Akzo Nobel N.V. | Carrier system against GnRH |
US5403586A (en) * | 1989-08-25 | 1995-04-04 | Biotechnology Australia Ptl Ltd. | LHRH-TraTp fusion proteins |
DE4003944A1 (de) * | 1990-02-14 | 1991-08-22 | Gerhard Dr Prinzhaus | Synthetisches polypeptid: lh - rh -- klh (konjugat aus luteinisierendem hormon-releasing hormon und dem keyhole limpet haemocyanin |
EP0501882A2 (fr) * | 1991-03-01 | 1992-09-02 | Rhone Merieux | Procédé d'immunoneutralisation anti-LHRM des animaux domestiques mâles non castrés et peptide pour cela |
WO1992015330A1 (fr) * | 1991-03-01 | 1992-09-17 | Rhône Merieux | Procede d'immunoneutralisation anti-lhrh des animaux domestiques males non castres et peptide pour cela |
EP0501882A3 (en) * | 1991-03-01 | 1992-10-14 | Rhone Merieux | Method of anti lhrh immunoneutralization of non-castrated male animals and peptide therefore |
US5573767A (en) * | 1991-03-01 | 1996-11-12 | Societe Anonyme | Method for improving the organoleptic qualities of the meat from uncastrated male domestic animals, vaccines which are usable in this method, new peptide, in particular for producing these vaccines and vaccination kit relating thereto |
US5955080A (en) * | 1991-07-26 | 1999-09-21 | Commonwealth Scientific And Industrial Research Organisation | Self-adjuvanting peptide vaccine delivery system and production thereof |
EP0596959A1 (fr) * | 1991-07-26 | 1994-05-18 | Commonwealth Scientific And Industrial Research Organisation | Systeme d'administration de vaccin peptidique produisant son propre adjuvant et production du vaccin |
EP0596959A4 (fr) * | 1991-07-26 | 1995-02-22 | Commw Scient Ind Res Org | Systeme d'administration de vaccin peptidique produisant son propre adjuvant et production du vaccin. |
EP0578293A1 (fr) * | 1992-06-18 | 1994-01-12 | Akzo Nobel N.V. | Protéine frangée (fimbrilline-P) de E.Coli comme porteur immunogène contre GnRH |
US5684145A (en) * | 1992-06-18 | 1997-11-04 | Akzo Nobel N.V. | Carrier system against GNRH |
US6303123B1 (en) | 1994-01-27 | 2001-10-16 | Aphton Corporation | Methods for the treatment of hormone-dependent tumors with immunogens against gonadotropin releasing hormone |
US6132720A (en) * | 1994-01-27 | 2000-10-17 | Aphton Corp. | Immunogens against gonadotropin releasing hormone |
US5688506A (en) * | 1994-01-27 | 1997-11-18 | Aphton Corp. | Immunogens against gonadotropin releasing hormone |
WO1998027111A1 (fr) * | 1996-12-17 | 1998-06-25 | Centro De Ingenieria Genetica Y Biotecnologia. (Cigb) | Vaccin pour l'immuno-castration reversible des mammiferes |
FR2763244A1 (fr) * | 1997-05-14 | 1998-11-20 | Pasteur Merieux Serums Vacc | Composition vaccinale multivalente a porteur mixte |
WO1998051339A1 (fr) * | 1997-05-14 | 1998-11-19 | Pasteur Merieux Serums & Vaccins | Composition vaccinale multivalente a porteur mixte |
US7862823B1 (en) | 1997-05-14 | 2011-01-04 | Sanofi Pasteur S.A. | Multivalent vaccine composition with mixed carrier |
US8029798B2 (en) | 1997-05-14 | 2011-10-04 | Sanofi Pasteur S.A. | Multivalent vaccine composition with mixed carrier |
EP1007084A1 (fr) * | 1997-07-09 | 2000-06-14 | Csl Limited | Composition immunogene de lh-rh et procedes concernant cette composition |
EP1007084A4 (fr) * | 1997-07-09 | 2003-07-02 | Csl Ltd | Composition immunogene de lh-rh et procedes concernant cette composition |
EP2196213A1 (fr) * | 1997-07-09 | 2010-06-16 | Pfizer Inc. | Composition immunogène de LHRH et procédés concernant cette composition |
EP3466441A3 (fr) * | 2012-03-09 | 2019-06-19 | Nord University | Procédés immunologiques d'inhibition de la maturation gonade |
WO2020051656A1 (fr) * | 2018-09-12 | 2020-03-19 | Ouro Fino Saúde Animal Ltda | Polypeptide chimérique antigénique, construction génétique et composition antigénique pour l'immunocastration de mammifères non humains |
Also Published As
Publication number | Publication date |
---|---|
DK114588A (da) | 1988-05-03 |
FI880954A0 (fi) | 1988-03-02 |
EP0274496A1 (fr) | 1988-07-20 |
DK114588D0 (da) | 1988-03-03 |
NZ220932A (en) | 1990-05-28 |
FI880954A (fi) | 1988-03-02 |
JPH01500900A (ja) | 1989-03-30 |
EP0274496A4 (fr) | 1989-12-19 |
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