WO2000020868A1 - Screen for risk for gastric adenocarcinoma - Google Patents
Screen for risk for gastric adenocarcinoma Download PDFInfo
- Publication number
- WO2000020868A1 WO2000020868A1 PCT/US1998/020820 US9820820W WO0020868A1 WO 2000020868 A1 WO2000020868 A1 WO 2000020868A1 US 9820820 W US9820820 W US 9820820W WO 0020868 A1 WO0020868 A1 WO 0020868A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cells
- gastric
- biopsies
- spasmolytic
- spem
- Prior art date
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57446—Specifically defined cancers of stomach or intestine
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/575—Hormones
Definitions
- Gastric cancer worldwide is the second highest cause of death from cancer. While rates of gastric cancer have been decreasing in the United States, gastric cancer prevalence remains high in other parts of the world, especially in Asia. Presently, patients in endemic regions for gastric center, especially Japan and China, undergo screening b ⁇ upper endoscopy. There is no alternative at present for discovery of early cancer other than by endoscopy. Grading of these endoscopies is based solely on hematoxylin and eosin staining. There are no prognostic markers that could indicate those at risk for future cancer development. Therefore, while biopsies can reveal early gastric cancers, patients in high risk regions who are initially negative by endoscopy must be followed with endoscopies to rule out future developments.
- spasmolytic peptide It has been determined that a specific metaplastic lineage that contains immunoreactivity for a trefoil polypeptide, spasmolytic peptide, is associated with and gives rise to the vast majority of human adenocarcinomas. The identification of this Spasmolytic Polypeptide
- SPEM Expressing Metaplasia
- hSP Spasmolytic peptide
- antibodies or nucleic acid probes hybridizing to mRNA encoding hSP can be used for rapid detection of the cells in tissue biopsies.
- the antibodies can also be used in serological tests for screening and following patients at risk for gastric cancer. In combination with evidence of previous or present invention with H. pylori, the tests are predictive of the liklihood of developing adenocarcinoma.
- Metaplastic cell lineages arising in response to chronic injury are precursors for the evolution of dysplasia and adenocarcinoma.
- This sequence is well characterized in the case of the Barrett's epithelium, a columnar specialized intestinal metaplasia in the distal esophagus of patients with esophageal reflux ( ⁇ aggitt, R.C. Hum. Pathol. 25, 982-993 (1994)).
- intestinal metaplasia has been associated with gastric adenocarcinoma (Dixon, et al. Am. J. Surg. Pathol. 20, 1161-1181 (1996); Filipe et al. Int. J. Cancer. 57, 324-329 (1994)); Correa, P.
- gastric mucosal biopsies are fixed in formalin and embedded in paraffin. Microtome sections of tissues are then stained with hematoxylin and eosin. These stained slides are examined for loss of parietal cells (oxyntic atrophy), ulceration, inflammatory infiltrates, and alterations in cell lineages including increased numbers of surface mucous cells (foveolar hyperplasia) , the presence of goblet cells (intestinal metaplasia), as well as dysplasia and adenocarcinoma.
- Dysplasia and adenocarcinoma ar judged by changes in nuclear morphology, loss of cytoplasmic space, loss of polarity and invasion of submucosa or vasculature. Brunner's glansds are not present in the normal stomach but can be observed in the duodenum.
- hSP Human spasmolytic peptide
- the patterns and timing of the expression of the trefoil peptides are different from each other. It is though that S2 plays an important rle in the proliferation of intestinal epithelial cells during the acute phase of mucosal ulceration, whereas ITF may be involved in differentialtion of the cells, particularly to form goblet cells, during the recovery phase of acute colitis. (Itoh, et al. , Biochem. J. 318(Pt 3), 939-944 (1996)). Immunostaining for SP in the intact mucosa has been determined to be confined to the mucous neck cells, but following exposure to stress it was significantly enhanced and occurred also in the cells of the basal region of gastric glands, as reported by Konturek, et al., Regul.
- SP is a marker of metaplastic cells having a morphology similar to those of Brunner's gland cells. These cells can be identified by histological examination. However, the identification of SPEM with spasmolytic peptide immunostaining is easier, more sensitive and rapid. Therefore, detection of metaplastic cells expressing SP provides a means for identification of those at risk who would need further follow-up. Furthermore, since the SPEM lineage is often extensive, quantitation of serum spasmolytic polypeptide levels by either radioimmunoassay or ELISA should be useful to stratify patients at risk and provide a serological method for identifying and following patients at risk for developing adenocarcinoma.
- SPEM can be detected using antibodies or antibody fragments prepared by standard techniques. The studies described in the examples were performed with a mouse monoclonal IgM anti-human spasmolytic polypeptide developed by Drs. Richard Poulsom and Nicholas Wright at the Imperial Cancer Research Fund, London, UK. Antibodies specifically directed towards utility in RIA and ELISA have also been developed. Either monoclonal or polyclonal antibodies can be used. Antibodies can be labelled using any detectable marker, including radiolabels, fluorescent labels, dyes, enzyme-chromogenic substrate systems, and other means commercially available.
- SPEM can also be detected using nucleic acid probes which hybridize under standard hybridization conditions, as described for example, by Maniatis, et al. Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Laboratory), to mRNA encoding SP. These can be labeled using standard labelling techniques for detection of bound nucleic acid. Alternatively, or in addition, other markers of these cells can be used to screen for the presence of SPEM in gastric tissue biopsies. For serological assay of SP, serum would be obtained from fasting patients. SP levels would be determined using either radioimmunoassay or enzyme-linked immunoassay. A standard curve would be used for known amounts of recombinant SP (Lars Thimm, Novartis Corporation).
- H. pylori is known to be associated with an increased incidence of adenocarcinoma of the stomach, efficacy of screening can be further enhanced by testing for previous or present H. pylori infection.
- H. pylori infestion would be determined by either CLO test at the time of biopsy or H. pylori serology with the same sample used for SP determination.
- SP detection such as immunohistochemistry
- endoscopic biopsies as well as brushings obtained either through endoscopy or blind per oral intubation.
- gastric cancer screening especially in Asian countries, is a major focus for clinical care.
- the only impact of medicine on gastric cancer is through early detection of low grade tumors and early resection for cure.
- High grade tumors have uniformly dismal prognosis with median survival less than one year. No significant effect of adjuvant chemotherapy has been noted.
- the addition of spasmolytic polypeptide immunostaining of biopsies and the identification of SEEM provides a means for identifying those at risk for developing cancer in the future.
- spasmolytic peptide serology should provide a blood test for identification of those at risk.
- the use of spasmolytic polypeptide immunostaining could significantly decrease the number of screening endoscopies, focus screening endoscopies, and, through serology testing, facilitate screening of large populations a risk in Asia and other countries with high cancer incidence.
- Example 1 Detection of SP levels in gastric fundic biopsies. Ten fundic biopsies from H. pylori negative patients and 17 biopsies from patients with H. pylori colonization in the fundus were examined. All biopsies demonstrated ⁇ /K-ATPase-immunostaining parietal cells and contained no gastrin-immunoreactive cells, confirming the authenticity of fundic mucosal biopsies. In all of the H. pylori- infected biopsies specimens, the presence of H. pylori was confirmed by Giemsa staining of tissue sections. The histological characteristics of these biopsies are summarized in Table I.
- Foveolar hyperplasia was assessed in sections stained for pS2 as the presence of surface cells in greater than 25 % of the gland length.
- Oxyntic atrophy was assessed in sections stained for H/K-ATPase as a decrease in parietal cell density to less than half the gland length.
- Mononuclear infiltrates were assessed in H&E stains.
- Hp H. pylori
- Gastric fundic biopsies were stained for SP. 10 ⁇ m sections from paraffin-embedded biopsies of fundic mucosa from H. pylori uninfected and H. pylon infected patients were stained for hSP with a monoclonal murine anti-HSP Elia, et al. Histochem. J. 26, 644-647 (1994)). Dewaxed paraffin sections were blocked with 5 % goat serum in phosphate buffered saline and then incubated with anti-hSP (1:50) for one hour at room temperature.
- H. pylori The presence of H. pylori was confirmed in all biopsies with Giemsa staining. All biopsies demonstrated immunoreactive parietal cells without the presence of G-cells). In non-infected patients, staining was confined to normal appearing mucous neck cells. In H py lori- infected patients, nodular aggregates of hSP-staining cells with the characteristics of Brunner's gland cells (SPEM) were observed in 65 % of biopsies. No SPEM was observed in fundic biopsies from non- infected patients.
- SPEM Brunner's gland cells
- Example 2 Association of SPEM with Gastric Adenocarcinomas. Since chronic H. pylori infection is associated with the development of adenocarcinoma, archived tissues from resected gastric adenocarcinomas in patients were examined. All of the resections were for tumors of the fundus or fundal/antral border.
- Adjacent sections were stained with anti-PSTI (1 :200) with staining as described above except that sections underwent retrieval using citrate buffer in a microwave and horseradish peroxidase- conjugated secondary antibodies were used for detection with diaminobenzidine chromogen. While distinct PSTI immunoreactive mucous neck cells were observed within the mucosa, the SPEM lineage cells at the right side of the section showed no PSTI immunoreactivity. Adjacent sections were also stained with anti-PS2 (1 :50) as described above. While pS2 immunostaining was observed in regions of surface cell foveolar hyperplasia, SPEM cells did not stain for pS2.
- Toluidine blue staining of a 0.5 ⁇ m section of glutaraldehyde fixed tissue from a resection specimen demonstrates the numerous granules in SPEM cells. Note the presence of residual parietal cells in some SPEM-containing glands, confirming the presence of the lineage in fundic mucosa. Electron microscopic examination demonstrates the presence of multiple mucous granules within the SPEM cells along with the characteristic expanded apical membrane surface without microvilli.
- ITF Intestinal Trefoil Factor
- the SPEM lineage was observed in apposition with both dysplastic lineages as well as adenocarcinoma.
- hSP staining was observed in both SPEM as well as in contiguous regions of dysplasia in 49% of resection samples. Immunoreactivity was detectable in a region of adenocarcinoma adjacent to the hSP-immunoreactive cells.
- dysplastic epithelial cells appear to be contiguous with SPEM cell-containing glands.
- hSP immunoreactivity was present in cells within regions of severe dysplasia or carcinoma in situ.
- regions of SP- expressing dysplastic cells were noted extending from regions of SPEM lineage. While regions containing the SPEM lineage did not show significant labelling with Ki-67 and PCNA antibodies, the dysplastic areas showed prominent nuclear labelling in addition to characteristic changes in cell morphology. These results indicate that the SPEM lineage may represent a metaplastic precursor for the development of dysplasia and adenocarcinoma. Previous investigations have focused on the association of Type III intestinal metaplasia with the development of adenocarcinoma. Despite this association, it is less clear that neoplastic cells actually arise from areas in goblet-cell containing intestinal metaplasia.
- intestinal metaplasia and foveolar hyperplasia are often conspicuous in atrophic gastritis in association with H. pylori infection
- gastric adenocarcinomas usually develop deep in the glands as nodular lesions that underlie the regions of intestinal metaplasia or foveolar hyperplasia.
- the identification of the SPEM lineage supports the proposal that this metaplastic candidate precursor is located in proximity with the putative site of neoplastic transition.
- hSP itself might contribute to the development of adenocarcinoma.
- hSP has generally been associated with cytoprotection in the gastric mucosa (McKenzie, et al. , (1997); Tanaka, et al. Am. J. Physiol. 272, G1473-G1780 (1997); Babyatsky, et al. Gastroenterology. 110, 489-497 (1996)).
- Homologous deletion of the gene for the gastric surface cell trefoil protein pS2 leads to the development of intramucosal carcinomas (Lefebvre, et al. Science 274, 259-262 (1996)), however the lineage responsible for these lesions is unclear.
- Esophageal adenocarcinoma develops from dysplastic transition within the metaplastic columnar Barrett's epithelium in the distal esophagus (Reid, et al. Gasronenterology. 102, 1212-1219 (1992); Miros, et al. Gut. 32, 1441- 1446 (1991)). As with the SPEM lineage in the stomach, the Barrett's epithelium shows low proliferative indices. Dysplasia and adenocarcinoma only develop in a minority (10-15 %) of patients.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Pathology (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- General Physics & Mathematics (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Oncology (AREA)
- Hospice & Palliative Care (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
Claims
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA002345663A CA2345663A1 (en) | 1998-10-01 | 1998-10-01 | Screen for risk for gastric adenocarcinoma |
JP2000574935A JP2002526776A (en) | 1998-10-01 | 1998-10-01 | Screening for risk of gastric adenocarcinoma |
AU96810/98A AU765082B2 (en) | 1998-10-01 | 1998-10-01 | Screen for risk for gastric adenocarcinoma |
EP98950883A EP1121597A1 (en) | 1998-10-01 | 1998-10-01 | Screen for risk for gastric adenocarcinoma |
PCT/US1998/020820 WO2000020868A1 (en) | 1998-10-01 | 1998-10-01 | Screen for risk for gastric adenocarcinoma |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/US1998/020820 WO2000020868A1 (en) | 1998-10-01 | 1998-10-01 | Screen for risk for gastric adenocarcinoma |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2000020868A1 true WO2000020868A1 (en) | 2000-04-13 |
Family
ID=22268006
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1998/020820 WO2000020868A1 (en) | 1998-10-01 | 1998-10-01 | Screen for risk for gastric adenocarcinoma |
Country Status (5)
Country | Link |
---|---|
EP (1) | EP1121597A1 (en) |
JP (1) | JP2002526776A (en) |
AU (1) | AU765082B2 (en) |
CA (1) | CA2345663A1 (en) |
WO (1) | WO2000020868A1 (en) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2003050500A1 (en) * | 2000-05-10 | 2003-06-19 | Lankenau Institute For Medical Research | Early diagnosis of cancer or precancerous conditions by leakage of signature peptides and carbohydrates into the bloodstream |
US7538082B2 (en) | 2001-04-24 | 2009-05-26 | The General Hospital Corporation | Methods and compositions for treating oral and esophageal lesions |
USRE41028E1 (en) | 1996-04-12 | 2009-12-01 | The General Hospital Corporation | Treating eye disorders intestinal trefoil proteins |
WO2012170478A2 (en) * | 2011-06-06 | 2012-12-13 | The University Of North Carolina At Chapel Hill | Methods and kits for detecting adenomas, colorectal cancer, and uses thereof |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1996015456A1 (en) * | 1994-11-16 | 1996-05-23 | Locus Genex Oy | Method for screening the risk of gastric cancer |
US5567594A (en) * | 1991-04-26 | 1996-10-22 | Enteron, L.P. | Methods and compositions for the detection and treatment of diseases associated with antigens of microorganisms |
DE19629938C1 (en) * | 1996-07-24 | 1997-11-27 | Gsf Forschungszentrum Umwelt | Monoclonal antibodies specific for mutated E-Cadherin peptide sequences |
US5783416A (en) * | 1993-01-21 | 1998-07-21 | Novo Nordisk A/S | Human spasmolytic polypeptide in glycosylated form |
-
1998
- 1998-10-01 JP JP2000574935A patent/JP2002526776A/en active Pending
- 1998-10-01 CA CA002345663A patent/CA2345663A1/en not_active Abandoned
- 1998-10-01 AU AU96810/98A patent/AU765082B2/en not_active Ceased
- 1998-10-01 EP EP98950883A patent/EP1121597A1/en not_active Withdrawn
- 1998-10-01 WO PCT/US1998/020820 patent/WO2000020868A1/en active IP Right Grant
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5567594A (en) * | 1991-04-26 | 1996-10-22 | Enteron, L.P. | Methods and compositions for the detection and treatment of diseases associated with antigens of microorganisms |
US5783416A (en) * | 1993-01-21 | 1998-07-21 | Novo Nordisk A/S | Human spasmolytic polypeptide in glycosylated form |
WO1996015456A1 (en) * | 1994-11-16 | 1996-05-23 | Locus Genex Oy | Method for screening the risk of gastric cancer |
DE19629938C1 (en) * | 1996-07-24 | 1997-11-27 | Gsf Forschungszentrum Umwelt | Monoclonal antibodies specific for mutated E-Cadherin peptide sequences |
Non-Patent Citations (2)
Title |
---|
SCHMIDT, P. H. (1) ET AL: "Association of an aberrant spasmolytic peptide -expressing cell lineage with gastric adenocarcinoma in humans.", GASTROENTEROLOGY, (APRIL 15, 1998) VOL. 114, NO. 4 PART 2, PP. A673. MEETING INFO.: DIGESTIVE DISEASE WEEK AND THE 99TH ANNUAL MEETING OF THE AMERICAN GASTROENTEROLOGICAL ASSOCIATION NEW ORLEANS, LOUISIANA, USA MAY 16-22, 1998 AMERICAN GASTROENTEROLO, XP002107070 * |
THEISINGER B ET AL: "EXPRESSION OF THE BREAST CANCER ASSOCIATED GENE PS2 AND THE PANCREATIC SPASMOLYTIC POLYPEPTIDE GENE HSP IN DIFFUS TYPE OF STOMACH CARCINOMA.", EUR J CANCER, (1991) 27 (6), 770-773. CODEN: EJCAEL. ISSN: 0959-8049., XP002107071 * |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
USRE41028E1 (en) | 1996-04-12 | 2009-12-01 | The General Hospital Corporation | Treating eye disorders intestinal trefoil proteins |
WO2003050500A1 (en) * | 2000-05-10 | 2003-06-19 | Lankenau Institute For Medical Research | Early diagnosis of cancer or precancerous conditions by leakage of signature peptides and carbohydrates into the bloodstream |
US7538082B2 (en) | 2001-04-24 | 2009-05-26 | The General Hospital Corporation | Methods and compositions for treating oral and esophageal lesions |
WO2012170478A2 (en) * | 2011-06-06 | 2012-12-13 | The University Of North Carolina At Chapel Hill | Methods and kits for detecting adenomas, colorectal cancer, and uses thereof |
WO2012170478A3 (en) * | 2011-06-06 | 2013-04-25 | The University Of North Carolina At Chapel Hill | Methods and kits for detecting adenomas, colorectal cancer, and uses thereof |
Also Published As
Publication number | Publication date |
---|---|
EP1121597A1 (en) | 2001-08-08 |
JP2002526776A (en) | 2002-08-20 |
AU9681098A (en) | 2000-04-26 |
AU765082B2 (en) | 2003-09-11 |
CA2345663A1 (en) | 2000-04-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US6372439B2 (en) | Screen for gastric adenocarcinoma | |
JP5959579B2 (en) | Gastric cancer detection marker | |
Aikou et al. | Tests for serum levels of trefoil factor family proteins can improve gastric cancer screening | |
JP3774196B2 (en) | Abnormal cell growth | |
US8470544B2 (en) | Detection of dysplastic or neoplastic cells using anti-MCM3 antibodies | |
WO2010047448A1 (en) | Diagnostic kit of colon cancer using colon cancer related marker,and diagnostic method thereof | |
US20110053156A1 (en) | Small cell lung carcinoma biomarker panel | |
DK2678446T3 (en) | Presence of erg gene rearrangements and protein overexpression in low-malignant pin (lg-pin) in prostate biopsies | |
Sato et al. | Gastric carcinoid tumors without autoimmune gastritis in Japan: a relationship with Helicobacter pylori infection | |
JP2004532390A (en) | PIN1 as a marker for prostate cancer | |
KR20180011725A (en) | Biomarker for diagnosis of extrahepatic bile duct carcinoma intrahepatic bile duct carcinoma, or gallbladder carcinoma | |
AU765082B2 (en) | Screen for risk for gastric adenocarcinoma | |
EP1552311B1 (en) | Method for detecting a risk of acid related disease in an individual | |
Kusamura et al. | Expression of p53, c-erbB-2, Ki-67, and CD34 in granulosa cell tumor of the ovary | |
KR101849699B1 (en) | Composition for diagnosing or prognosising cancer comprising fibronectin protein positive exosome | |
Basso et al. | Serum antibodies anti‐H. pylori and anti‐CagA: A comparison between four different assays | |
US8252542B2 (en) | Serpin B 13 as a marker for squamous cell carcinoma of the lung | |
Maeda et al. | Blood tests for asbestos-related mesothelioma | |
Wada et al. | Relationship between Helicobacter pylori tyrosine-phosphorylated CagA-related markers and the development of diffuse-type gastric cancers: a case-control study | |
JP2019158613A (en) | Detection of placental site trophoblastic tumor (pstt) using laeverin | |
JP2023177246A (en) | Method of predicting prognoses of breast cancer patient | |
Qin et al. | Antibody array-based proteomic screening of novel biomarkers in malignant biliary stricture | |
WO2003029826A1 (en) | Method for detecting a cancer or precancerous condition | |
WO2018048258A1 (en) | Use of ard1 as marker for measuring prognosis of or diagnosing liver cancer | |
EP3124974A1 (en) | Method for the normalization of immunological tests and kits for performing such tests |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AU CA JP |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
WWE | Wipo information: entry into national phase |
Ref document number: 96810/98 Country of ref document: AU |
|
ENP | Entry into the national phase |
Ref document number: 2345663 Country of ref document: CA Ref country code: CA Ref document number: 2345663 Kind code of ref document: A Format of ref document f/p: F |
|
ENP | Entry into the national phase |
Ref country code: JP Ref document number: 2000 574935 Kind code of ref document: A Format of ref document f/p: F |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1998950883 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 1998950883 Country of ref document: EP |
|
WWW | Wipo information: withdrawn in national office |
Ref document number: 1998950883 Country of ref document: EP |
|
WWG | Wipo information: grant in national office |
Ref document number: 96810/98 Country of ref document: AU |