Number 2 2002
Dr Chris Corless
will deliver a lecture entitled
Dr Corless is a Staff Pathologist and Associate Professor of Pathology at Oregon Health and Science University and its associated hospital. He has a long-standing interest in gastrointestinal, urological and transplantation pathology.
He has recently been closely involved with developments in the understanding of gastrointestinal stromal tumours (GISTs) and C-KIT, and the relevance of this to treatment with Glivec (STI-571).
His lecture will be delivered on Saturday June 1, at 4.45pm His visit is being sponsored by Novartis.
Other invited speakers are Dr Robert Petras on Saturday and Dr David Owen on Sunday
29th Annual Scientific Meeting Friday 31 May to Sunday 2nd June
Companion Meetings Programme
Friday 31 May, 2002.
INTERNATIONAL SOCIETY OF GYNAECOLOGICAL PATHOLOGISTS
Convenor: Dr Richard Jaworski
Muscle Tumours of the Uterus: A Selected Review and Recent Advances"
This will be followed
by 5 interesting gynaecological pathology cases.
Convenor: Dr Victor J Ojeda
Meeting Room 2,
Lecture: Frozen Sections
and Neurosurgical Biopsies
2. Slide Seminar
Convenor: Dominic Wood
Chairperson: Geoffrey Strutton
8:55 - 9:20
9:20 - 9:40
9:40 - 10:40
A nodule on the ear
A nodule on the scalp
– Superficial Angiomyxoma
infection masquerading as a vasculitis
Are monoclonal and mixed types distinct histologically?
Unusual nodules on
the lower limbs – Phlebitic tuberculid
FACULTY OF ORAL PATHOLOGY
Convenor: Dr Michael Aldred
8:55 - 10:40 am
SOFT TISSUE TUMOURS
Convenor: Dr Richard
This will be followed by 5 interesting Soft Tissue Tumour cases
Convenor: Dr David Cohn
11:00 – 11:20 am
11:25 – 12:05pm
12:10 – 12:40 pm
Convenor: Dr Jenny
11:00 – 12:45
Convenor: Dr Gelareh Farshid Main Auditorium
2:00 – 3:34 pm
Convenors: Dr Judy Bligh and Dr Ann Finney
2.00 - 3.45pm,
1. Lecture: FNA
of the Liver
2. Lecture: FNA
of the Pancreas
3. Lecture: Bile
Duct Brushings, an overview
4. Lecture: A
Practical approach to Bile Duct Brushings
5. Case presentations:
Images from the three case presentations can be viewed prior to the meeting in the member’s section of the Australasian IAP web site www.iap-aus.org.au (user name: iapmember and password: rokitansky)
2:00 – 3.45 pm
Lecture: The Laboratory
Investigation of Genetic Metabolic Disorders Presenting with Liver Disease
Liver Diseases and the Anatomical Pathologist
6 cases for diagnosis
Carditis - Professor P Bhathal
Gastritic Observations - Dr John Pedersen
Intra-epithelial lymphocytosis in architecturally normal small intestinal biopsy specimens - Ian Brown
Inflammatory Bowel Disease:Uncommon Patterns and diagnostic pitfalls - Andew Clouston
Unfortunately there will be no Lymphoma Club Meeting this year.
Convenor: Dr Geoff
There will be a short presentation from Dr Hema Samaratunga discussing work she did with Jonathon Epstein on low-grade urothelial neoplasms and which she then presented at the Chicago IAP. This will be followed by a series of 7 slides.
VISIT OF I.A.P. PRESIDENT ELECT
Professor Shinichiro Ushigome, Emeritus Professor, Jikei University, Tokyo, and Past President of the Japanese Division will be attending our ASM.
Professor Ushigome will be President of the I.A.P. for our Congress in 2004. This visit will allow him to meet members of the Division and to inspect the Congress facilities in Brisbane.
Our Division's website can be accessed at: http://www.iap-aus.org.au or http://iap-aus.org.au
Within the site you will find information about the Division, this year's Annual Scientific Meeting and information about registering on-line, newsletters, information about other important pathology meetings, links to other sites of pathological interest, information about contacting the secretariat and information about becoming a member.
I am developing a Member's section on the website. In this area you will find interesting educational cases. To access the Member’s site section you will need to know the username and password.
User name: iapmember
You need to enter these in lower case.
The Division's website is constantly being updated so remember to REFRESH the page in your browser. I would like to hear from members as to what they would like to see put on the website and how I can improve it. You can contact me at :
Phone: (02) 9845 6222
Kindly check you records as to whether your subscriptions for Membership of the IAP are up to date.As a financial member you are entitled to discount attending the Annual Scientific Meeting.
Payments should be sent direct to the IAP office at 207 Albion Street, Surry Hills NSW 2010. (Not enclosed with payment for the ASM to Hoteliers International).
Memories of the 2001 Meeting
Noel Gordon-Glassford (Auckland), Myfanwy Plunkett (Perth), Kerry Knapp (Perth)
Dianne Payton (Brisbane) and Suzanne Arbuckle (Sydney)
Frances Petrey (Melbourne), Alison Skene (Melbourne), Leonard Wu (Melbourne)
Hoteliers International have generously offered to process registrations for the ASM this year along with taking hotel bookings. Please utilise Hoteliers service and forward booking form as included in the registration brochure
Sponsorship Donations for Congress 2004
The Third I.A.P. Asia Pacific Meeting
The Third I.A.P. Asia Pacific Meeting is to be held in Bangkok January 21 - 23, 2003.
Delegates and speakers are required.
This is a combined meeting of all the Divisions in the Asia Pacific Region.
These include Thailand, India, Japan, Korea, Hong Kong, Indonesia and Australia and New Zealand.
For more information
of an Australian
Morey, A.S. Field Anatomical Pathology,
Since December 2001, Australian women with metastatic breast carcinoma have had access to government-funded Herceptin therapy if their tumour has been demonstrated to score 3+ on HER-2 immunohistochemistry (IHC) (regardless of antibody used) or is positive for HER-2 gene amplification by FISH. A centralised laboratory for HER-2 FISH testing funded by Roche on a per-test basis has been established in our University teaching hospital Anatomical Pathology department for assessment of cases with equivocal IHC. We have previously correlated tumour type and grade with IHC and FISH results on 30 cases from our laboratory (1), and here extend this analysis to the first 263 cases (the majority with 2+ IHC) referred from elsewhere in Australia.
Considerable variation in hybridisation efficacy has been noted between paraffin-embedded tissue blocks from different laboratories, most likely due to different fixation and processing protocols. This has resulted in an 18.6% repeat rate and a 14.4% non-diagnostic rate. As many of the blocks are archival, and a retrospective analysis of processing variables difficult, we took surplus fresh material from two breast tumours (Grade 3 ductal carcinomas, subsequently shown to be IHC 3+ and FISH positive), and assessed a variety of fixation strategies with the aim of detecting procedures deleterious to FISH analysis.
i) Analysis of referred specimens
The 263 referred cases came from over 40 different laboratories. Tumour type and grade was independently reviewed (A.S.F) on H&E stained sections then compared with IHC and FISH results. 219 cases were primary tumours and 44 were nodal deposits or distant metastases. Outside IHC staining had been performed using several different primary antibodies and various protocols including the HercepTest (Dako). If IHC had not been performed prior to referral, in 64 cases this was performed using an in-house assay employing Dako polyclonal antibody A0485 (diluted 1:1000) on a Dako Autostainer after antigen retrieval in citrate buffer (pH6) for 40 mins at 95C. In preliminary experiments using antibody dilutions from 1:200 to 1:1600 on 19 tumours we found that this 1:1000 dilution most closely mimicked the result of the more expensive HercepTest (Dako). Detection was via the Vecta ABC Elite system. Staining was scored from 0 to 3+ according to HercepTest criteria.
FISH was performed on 4 micron paraffin sections using the Vysis Paraffin Pretreatment Reagent Kit and dual probe PathVysion HER-2 DNA Probe Kit, according to the manufacturer’s instructions. A positive control section was included in each batch. Signal was analysed using a Zeiss Axioscope II microscope with attached Axiocam digital camera and Axiovision software. Gene amplification was defined as a HER-2/chromosome 17 ratio greater than 2.
ii) Mock fixation experiments
Small samples of fresh breast carcinoma tissue were exposed to a range of fixation procedures; the first experiment involved comparison of HER-2 IHC and FISH results on tissue fixed in 10% neutral buffered formalin for 2 hr, 18 hr or 6 days. The second experiment involved comparison of IHC and FISH results on tissue samples exposed to: 4hr formalin fixation; freezing followed by 4hr formalin; microwave-aided (30mins at 60C) formalin fixation; delayed (by 2hr) formalin fixation; warm formalin fixation (60C, 18hr); and 2hr Carnoy's fixation followed by 2hr formalin. Similar variables have been represented among the cases referred to us, sometimes in combination. All results were digitally photographed at the same exposure under identical illumination. A subset of results is shown in Figure 1.
FISH RESULTS on 146 cases with “Scored” Outside IHC vs 64 CASES with In-House IHC (Dako A0485, 1:1000)
* includes borderline
FISH RESULTS BY TUMOUR GRADE
* includes 2 tubular carcinomas
# the amplified cases were an invasive papillary tumour, grade 2, and one of 3 pleomorphic lobular carcinomas, grade 2.
@ due to limited material or crush artefact.
Figure I: Subset of Results of Mock Fixation Experiments
DISCUSSION OF RESULTS
These data reinforce our previous suggestion:
(1) that triaging of cancers by type and grade could be used to focus HER-2 testing on tumours that are potentially amplified. Of the 141 assessable grade 3 ductal carcinomas referred with equivocal IHC, 44% were FISH+, whereas only 29% of the 52 assessable grade 2 ductal carcinomas and none of the 9 assessable grade 1 ductal/tubular carcinomas were FISH+. None of the 6 classical lobular carcinomas was positive, although one of three “pleiomorphic” lobular carcinomas showed low-level amplification. There was excellent correlation between in-house IHC 3+ and FISH+ (100% of assessable cases), although “external” 3+ IHC results showed slightly lower specificity (79%). All but two of the 37 IHC 1+ were FISH-negative (IHC slides were not reviewed); one of these showed borderline low-level amplification. The overall proportion of IHC 2+ cases found to be amplified (40%) is similar to recent reports
(2). Mock fixation studies demonstrated a surprising resistance of 3+ IHC and amplified HER-2 hybridisation signal to a range of "abusive" fixation regimens. Despite the fact that Vysis only guarantee successful FISH results for tissues fixed in 10% neutral buffered formalin for between 24 and 48 hours, in reality we have to deal with archival tissues fixed and processed in a variety of ways, often aimed at speeding up turn-around times. There is little available literature on the effects of such procedures on FISH efficacy. In general, FISH was more resilient to fixation variables than IHC, with most of the variables examined having minimal effect on FISH signal strength, however we noted a small diminution in hybridisation signal with prolonged fixation (Fig. 1b), and a marked loss of signal and increase in background autofluorescence in tissues exposed to Carnoy's fixative (containing chloroform and acetic acid) (Fig. 1c). Further questioning of referring laboratories supports the thesis that this "defatting" fixative may be responsible for a substantial proportion of our non-diagnostic cases. It is suggested that consideration be given to setting aside a small portion of each breast carcinoma at the time of receipt for "optimal" 24 hr formalin fixation and potential subsequent HER-2 IHC/FISH, regardless of the fixation and processing regimen pursued for the remainder of the specimen.
1). Field AS, Chamberlain N, Tran D, Morey A (2001) Suggestions for HER2/neu testing in breast carcinoma based on a comparison of immunohistochemistry and fluorescence in situ hybridization. Pathology, 33: 278-282.
2). Mass RD, Sanders C, Kasian C, et al. The concordance between the clinical trials assay and fluorescence in situ hybridization in the Herceptin pivotal trials. ASCO 36th Annual Meeting, May 2000, New Orleans. Abstract 291.
OF MATERIAL FOR HER-2 FISH TESTING
Please arrange for a paraffin block containing tumour tissue and a copy of the pathology report to be forwarded to:
FISH Laboratory Anatomical Pathology, L16 O'Brien Bldg St.Vincent's Hospital Victoria St., Darlinghurst NSW 2010
If a patient does not fulfill the above criteria, FISH will not normally be funded by Roche, but may still be arranged through the laboratory. Enquiries about this, or other aspects of the test should be directed to Dr Adrienne Morey or Dr Andrew Field on (02) 8382 2319.