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To find the best ways to screen for inherited diseases. To conduct quality research of an international stsuidard. To meet the needs of those who come to us for advice. To develop the most effective methods of counselling. To move towards treatment for genetic diseases.
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The genetics of brain and nerve diseases: how they occur in some families, what happens to the genes themselves, and developing new ways to treat them. Discovering genes which can cause deafness, blindness and other serious handicaps using Australian family resources. We are a recognised centre of excellence in genetics and have attracted to our staff many world class doctors, scientists, counsellors, nurses, administrators, and scholars.
We are one of Australia's top research centres. a World Health Organisation reference centre for human genetics. the premier training centre for clinical geneticists in Australia. the screening centre for all Victorian newborn babies. an international model for combined clinical and research efforts. the originator of the POSSUM/OSSUM computer system for the diagnosis of birth defect syndromes, which is now used by specialists in over 50 countries. the Australasian screening centre for genetic diseases of cell energy.
Our Research The body's use of copper, and the genetic diseases causing copper deficiency (Menkes disease) and copper toxicity (Wilson disease). The structure and function of the human chromosome; how chromosomes divide, and why this can go wrong (Down syndrome). The understanding, diagnosis and treatment of conditions affecting brain and muscle function in both childhood and adult life, including mitochondrial and metabolic disorders. Understanding embryo development and how it contributes to birth abnormalities of form (facial clefting, spina bifida and great vessel/cardiac defects) and cell differentiation (paediatric cancers like neuroblaistomas and leukaemiats). Somatic gene therapy: how to treat inherited diseases in better and more natural ways by using normal copies of the genes that do not work properly in ataxias, cystic fibrosis and other genetic disorders. Page ii
HOW CAN YOU HELP? \bu can help the Murdoch Institute reach its goal of helping every child to be bom healthy and with normal abilities through: ' bequests ' annual donations ' pledges over several years ' named scholarships or research gifts For further information, please telephone or write to: Professor Bob Williamson Director The Murdoch Institute Royal Children's Hospital Remington Road, Parkville 3052 Telephone: (61) 3 9345 5045 Fax:: (61)3 9348 1391 The Murdoch Institute is a registered charity. All donations over $2.00 are tax-deductible.
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Tine M^ufJoc 1 iinstitiULte
Director Board of Directors
Professor Bob Williamson
Victorian Clinical Genetics Service Board of Directors
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page
Open Day
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Maternal Serum Screening
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Mr L.G. Cox, Chairman Professor J. Angus Dr G.L. Barnes Mrs J. Calvert'Jones Mrs L. Cattermole Mr I. Davies Mr J.A. Fitzgerald
POSSUM
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Victorian Clinical Genetics Services
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Chromosome Research Group
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Trace Element Group
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Mr P Griffin
Gene Identification and Expression Group
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Gene Therapy
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Metabolic Research
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Embryology Group
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Mouse Model Unit
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Tissue Culture
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Epidemiology
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DNA Diagnostic Laboratory
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Metabolic Laboratory
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Newborn Screening Laboratory
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Cytogenetics Laboratory
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Genetic Counselling and Cancer Workshops
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Post Graduate Diploma in Genetic Counselling
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Appreciation to our Supporters
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Murdoch Institute Staff
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Victorian Clinical Genetics Service Staff
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List of Publications
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Financial Statements
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Mr J.S. Guest Mr W.H. Hodgson, Deputy Chairman Mrs A. McFarling Mrs J. Paterson Professor D.G. Penington Professor P.J. Smith Professor G.W. Tregear Professor R. Williamson
Finance Committee
Achievements and New Directions
Mr P.J. Griffin, Chairman Mr C.P. Abbott Mr L.G. Cox Mr D.T Craig Mr G.E. Heeley Mr D.E. Meikeljohn Mr 1. Miller Mr 1. Veal Mr L.G. Cox, Chairman Dr G.L. Barnes Professor H. Ekert Mr J.S. Guest Mr W.H. Hodgson Mr G.E. Heeley Dr L.J. Sheffield Professor R. Williamson
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AdiieTements an
JN ew .Directions
The Murdoch Institute is the msijor Australian Research Institute studying medical genetic problems in childhood. During 1996, the Institute's NH&MRC Block Grant was renewed following an assessment by an international team of experts appointed by the NH&MRC. The renewal of this funding is a true accolade from the Australian biomedical research community, as only four other Institutes share this kind of government support - the Walter and Eliza Hall, Florey, Baker and Garvan Institutes. We are pleased to report that in passing this close scrutiny we also obtained an increase in our level of support. The procedures were hard but fair, and we are delighted to acknowledge the support of all of our colleagues in our bid for renewal. The final report stated that a major component of our success is due to our concept of an integrated Institute which combines research, diagnostic and clinical efforts at the highest international standard. Although renewal of our Block Grant renewal was the most important event of 1996, we have many other things to celebrate. We share a grant from the Commonwealth Government of $13 million over seven years for a Collaborative Research Centre in Gene Discovery. This will be directed to identifying genes for common diseases both of children and adults. We have a special interest in genes causing eczema, blindness and deafness, which cause great difficulties for many thousands of young Australians. If the genes can be found, it will help with the development of new forms of treatment, which we hope will also aid our young Victorian biotechnology industry, including AMRAD, which is one of our partners in this venture. Government granting bodies are conservative and look backwards for a track record. We are therefore grateful for the ongoing support of our friends, the Brockhoff Foundation, the Miller family and the Murdoch family. Without them, we could not make the initiatives which are so necessary in a rapidly moving field. The gene therapy team, now sited in the Olive Miller laboratory, is funded by generous grants from these core supporters. One centrepiece of our work is the service we give to those who are ill with genetic
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disorders through the Victorian Clinical Genetics Services (VCGS). The clinical team has been expanded to six clinical fellows plus a further three doctors carrying out research full time in the Institute. The budget of the Victorian Clinical Genetics Services has been increeised by approximately 30% by the Department of Human Services, a recognition of the increasing importance of genetics in medicine. This is a remarkable tribute to our clinical team at a time when most budgets are (at best) static. We also thank the State government for agreeing to share the cost of refurbishing the clinical wing with the Murdoch family. When completed this will provide comfortable, modern, purpose-built clinical facilities. This is another example of “shared support", which seems to us to be an excellent way to combine government and charitable support for the best in health research and care. We are also proud to acknowledge the support of many patient support and charitable groups. These range from research into gene therapy for Friedreich's ataxia, cystic fibrosis, Menkes and Wilson diseases and Down syndrome through to studies on the effect of copper on young children and the development of human artificial chromosomes. Over $ 150,000 has been donated to support these research activities. We have also used some of this funding to expand our epidemiology group, which is conducting research into what people want from genetic clinics, and how they make use of what is on offer. In the past, most of our studies have concerned severely ill children, and this continues to be our main focus. Any society is best judged on how it treats the most vulnerable of its young. However, we are also spending more time with genetic conditions affecting adults, especially cancer and neurological disorders, in collaboration with our colleagues in major teaching hospitals such as Monash Medical Centre and the Peter MacCallum. The State Department of Human Services has provided $250,000 per annum to provide gene testing for colon cancer, jointly with the Royal Melbourne Hospital and Victorian Anti-Cancer Council. We must maintain an awareness of the ethics of all our activities. We discuss ethics, think ethics and teach ethics continuously, and have used a generous donation to appoint a Senior Research Associate to head a new unit carrying out research into the Ethics of the New Genetics. We are also mindful of the need for education, and have appointed a Senior Lecturer in Medical Genetics (through the
Laurie Cox and Bob Williamson
Open Day
The Murdoch Institute's first Open Day was held on April 24, 1996.
Department of Paediatrics, based in the Murdoch) and an outreach Education Officer. Their task is to ensure that full discussion surrounds all our activities, both in house and in the media, the profession, and, perhaps of most importance, in Victoria's schools. As part of our outreach effort, the Murdoch is also “home" to thirty or so support groups, which meet the needs of those with a particular genetic disability. These provide practical and emotional support and advocacy for our patients and their families.
On this day our new wing was formally opened by our patron and founder. Dame Elisabeth Murdoch. Supporters, donors, families, patients and collab' orators were able to visit the Institute and see its facilities, the laboratories and the clinical services in action.
Our staff continues to excel. Julian Mercer and Andy Choo have been appointed Associate Professors of the University of Melbourne, and many of our younger staff have won prizes at meetings for "best presentation". Our papers appear in the best journals including Cell, Nature Genetics, and Human Molecular Genetics. Our focus is not just to isolate genes, but to find out how they work, as this is of great clinical relevance. During the coming year, we hope to consolidate our research effort as well as our clinical and diagnostic services, to ensure that our contribution to Australia continues to be in the forefront of human genetics research and clinical practice.
Bob Williamson, Director
Laurie Cox, Chairman
Educational displays of the Institute's activities were viewed with interest. The refurbishment of the north west wing has created modern facilities in attractive surrounds for the cytogenetics, newborn screening and diagnostic laboratories. Barry Holt and his team transformed what was previously a jumble of play rooms, storage areas and junk rooms into a first rate laboratory, training facility and research area. We plan to make the Op en Day a regular event, with the next
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M^aternal S eiriinni
s creenmg
POSSUM
Every woman hopes for a healthy baby. Maternal serum screening for Down Syndrome and neural tube defects is one voluntary test that can be offered for this purpose. Commencing 1 st July 1996 the Victorian Clinical Genetics Services introduced a program offering pregnant women this blood test which can deter' mine if they are at increased risk of having a baby with Down Syndrome or a neural tube defect.
After 10 years of success and eight updates POSSUM is now preparing for a new leap forward. Under the leadership of John Marquet, the Computer Power Group (CPG) is preparing a system by which clinical geneticists, genetic counsellors and others members of the user^community can access Pictures of Standard Syndromes and Undiagnosed Malformations and data via the Internet.
Pregnant? A blood test during pregnancy can identify babie.s at risk ofN euralTube D efects or D own Syndroir e.
In V-rjcrii, around 1 ii Uitii-;;'1i,V/PK ;3’ P'-C y-'Sr!. "T ors: co;r. m oi; of che.'so in Sphi B iliid, ‘knri; PH-jv opft-.ru in d-.rt onneiior, Ci-y wh-nh r-.n ddc SO'; H'' d'lS nervcn contvLi'.q chp k'W'i' cf Lhfe r.'0'’iy,
A prototype POSSUM written in the Java language, running on Microsoft Windows using the data and video disc content of the latest version of POSSUM (Version 4.5) met with an enthusiastic response at the 1996 American Society of Human Genetics Annual Meeting in San Francisco.
I-Vrtori.crcuna L r, • onhs.' rinv^; i) ow;. t'yndrouio per yean. Ii :vn n-d.sr iiue'jw.ns! ‘.■iiv.biiiry, heart dpfeciis, and dilrcdlrisis-w in sifn; hn-.ring.
Future versions (under construction at CPG) will allow members of the POSSUM community to exchange data and pictures in an orderly way, and maintenance of POSSUM content should become quicker and simpler. JAVA will allow POSSUM to run in most computers, including the Apple Macintosh.
•A " .neC Tftsr Laboratory c?s6SJi ottr,' bbo-a .%!r; p&. 4 rjabstcmcjs h m obi-u:'. :;bod arc p eanur-sd, "A’ InfLnw aLbn. co..;r;;X'lliiia, ambtan:.'';' tc iiton^ryt cho ruarj.;:, orhiib b deaUio w ih any wouy ot |•.ono?m -iboii". tfio
Leonard Bonacquisto preparing serum samples for analysis
{
Ask your doctor about the M|IA»KIIPLE TEST",
Agnes Bankier (Team Leader) and Catherine Rose maintain and update the POSSUM picture and data bases with help from David Sillence (Sydney) and Moshe (Juan) Chemke (Israel). Dr Chemke has recently updated and merged the data of OSSUM (the spin'off system on skeletal dysplasias) into POSSUM.
The test we are using measures four substances in the mother's blood. After analysis in the laboratory a result is determined which is either "low risk" or "high risk". If there is a high risk, further tests such T as amniocentesis or ultrasound can be done to confirm whether the developing baby has Down Syndrome or a neural tube defect.
Marketing, distribution and customer support is managed by Anne Cronin and Henny Miller at the Murdoch Institute.
We recognise that the decisions regarding these tests can be stressful, and to provide support to those who need help the VCGS offers a comprehensive information and counselling service. Women are welcome to either phone us with their concerns, or they can arrange to see one of our genetic counsellors who can help them understand the implications of the test.
The POSSUM Team from left: Moshe Chemke, Anne Cronin, Gavin Arndt, John Marquet, Agnes Bankier, Kathy McNiff, Cathy Rose. Absent: Henny Miller and David Sillence
For news about POSSUM developments, visit the Web pages at http://vvww.rch.unimelb.edu.au/murdoch/possum.htm
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I
Victorian Clinica. Genetics S CFTices
There is a fresh excitement in the VCGS. The past year has seen the fruits of the tremendous energy and enthusiasm of Professor Bob Williamson, Director of the Murdoch Institute and Executive Director of the VCGS, with the arrival of new staff, new initiatives and the reality at last of redeveloping the clinical wing, which should provide the much needed accommodation for the VCGS. For some time we have wanted to recruit a doctor trained in the care of inborn errors of metabolism to run this service on a long term basis. We are pleased to welcome Dr Avihu Boneh, who takes up his post in March 1997 and will be in charge of the Metabolic Clinical Service. Metabolic physician Maureen Cleary will be returning to Manchester. We thank Maureen for her excellent care of our patients. The anticipated arrival of five new trainee Fellows in Clinical Genetics will revive an academic atmosphere, making it possible to engage not only in clinical service but also in clinical research. We welcome Frangois Bernier (Canada), Helen Heussler, a paediatrician on a 6 months fellowship, Ravi Savarirayan (South Australia), Stephen Robertson (New Zealand) and Simon Hauser who have joined Karen Dunn and Heidi Peters. We can look forward to a productive 1997.
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Counselling clinics for neurogenetic disorders are now being conducted by Mac Gardner and Margaret Olsen in conjunction with Elsdon Storey at the Alfred Hospital and at St Vincent's Hospital. Although a relatively new concept, these clinics are in high demand by adults who are either affected by inherited disease or have a genetic risk. In nearly all cases diagnosis will initially have been made by a neurologist. The clinics provide genetic diagnosis, genetic counselling and advice on the nature of the genetic Mac Gardner and Margaret Olsen studying an MRl inheritance and their future due to the diseaise. scan of a patient with an inherited brain disorder The spiralling link of genetics to cancers has created a high demand for counselling in the familydinked cancers at St Vincent's and the Royal Melbourne, and we foresee an expansion in the need for counselling in cystic fibrosis and haemophilia. Additional inherited cancer counselling clinics have been established by Mac Gardner at the Royal Melbourne Hospital, and Mary^Anne Young will be involved in the establishment of the family cancer clinic at the Peter McCallum Hospital. Further clinics are planned at other hospitals including the Austin Hospital. The VCGS has continued to provide a network of outreach clinics in country centres in Victoria, with Agnes Bankier now conducting counselling services for Tasmania.
The Post'Graduate Diploma Course in Genetic Counselling under the auspices of Melbourne University will enter its second year, with 14 students registered. Leslie Sheffield and Margaret Sahhar continue to coordinate the course. Our contribution to the training of medical students (Monash and Melbourne Universities), and medical and nursing colleagues hais been facilitated by the appointment of two very able people to the Murdoch Institute: Sylvia Metcalfe, Senior Lecturer in Medical Genetics, and Mary-Anne Aitken, Community Education.
Education and close liaison with a large number of patient support groups has continued under the guidance of Margaret Sahhar.
Our laboratory diagnostic services and newborn screening program have had a busy year. The newly established Maternal Serum Screening Program (for pregnant women to identify those at high risk of having a baby with Down syndrome) has more than met the projected targets in its first half year of operation. We are delighted to welcome back Robin Forbes to be coordinator of this program. Robin returns from Western Australia where she had worked for a number of years. The VCGS has been charged to provide counselling services for inherited colon cancer with dedicated funding from the Victorian government. The planned redevelopment of the clinical wing is a most exciting prospect. This will provide Margaret Sahhar and Leslie Sheffield
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Ck Fomosomrie Researc li G roMp
accommodation for the VCGS staff including much needed consulting rooms, patient waiting areas and meeting rooms. It is anticipated that this will be completed in the coming year. As chairman of the Clinical Genetics Steering Committee, Bob Williamson together with Agnes Bankier and Sue Forrest, assisted Barbara Mouy in producing a five year plan for the provision of clinical genetic services for the state of Victoria. This document has now been received by the Minister and has been adopted as part of the service plan for the VCGS.
With great anticipation we await the arrival in the coming year of the new Clinical Director of the VCGS. To this position we appointed Stephen Kahler (presently at Duke University, USA), a clinical geneticist and well known expert in laboratory diagnosis and management of metabolic disorders, with a keen interest in genetic education. This will free Agnes Bankier from her current administrative duties and enable her to lead the redevelopment of the POSSUM Project.
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Group Leader Scientific Officer Research Assistant Research Assistant Postdoctoral Scientist Postdoctoral Scientist Postdoctoral Scientist (till Nov '96) PhD Scholar (till Dec ’96) PhD Scholar PhD Scholar PhD Scholar PhD Scholar BSc Honours Student
Sue Forrest, Bob Williamson and Agnes Bankier
In July 1997, the VCGS will undertake a voluntary external review. The review panel chaired by Eric Haan (South Australia) includes Peter Harper (Cardiff) and Bridget Wilcken (New South Wales). We look forward to their review and comments on our services.
Agnes Bankier Clinical Director
Andy Choo Elizabeth Earle Kellie Tainton Danielle Irvine Michael Cancilla Richard Saffery Helen Trowell Desiree duSart Paul Kalitsis Damien Hudson Alyssa Barry Emily Howman Andrew MacDonald
Leslie Sheffield Director of Education 6- Training
Towards a Human Artificial Chromosome. Our research continues towards the construction of a human artificial chromosome. If we can make our own chromosomes we are well on the way to successful gene therapy. Genetic information is carried on genes within chromosomes. Sometimes this genetic information is wrong and leads to illness. Each normal cell has 46 chromosomes and our artificial chromosome would be introduced into cells as the 47th chromosome. While chromosome number 47 may be one thousand times smaller than any normal human chromosome, it will have the ability to carry corrected genes which could cure inherited gene defects. A number of different prototype artificial chromosomes have already been stitched together in the laboratory by our research group and we are presently testing these in cell culture. A basis of our research is studying the centromere of the chromosome at the time of cell division when each chromosome makes a copy of itself The chromosomes then utilise a special machinery to distribute the new chromosome copies equally between two newly formed cells. Our Page 10
studies have identified a snia.II segment of the chromosome that is responsible for the proper functioning of this machinery (the centromere), giving us the unique opportunity to undertake a research program to construct a human artificial chromosome.
found to bind a nuclear protein, which we have named pHC-1. This protein is a potentially important structural component of the human centromere and work is in progress to study and clone the gene encoding this protein using the "oneTybrid" system. The "one-hybrid" system is also being established in the laboratory for the analysis of another novel centromere-binding protein, pda, which we have previously reported to bind the a-satellite DNA of normal centromeres.
Centromere Protein Mutations. In another research area, we postulate that mutations (or damages) in the genes that control the chromosome distribution machinery can cause a catastrophic misdivision of chromosomes during cell division and result in fetal cell death during embryo development in pregnancy. By mutating two such genes in laboratory mice our results have so far demonstrated that mutations in one of these genes have indeed caused early fetal loss, but surprisingly, mutations in the second gene have so far not produced any ill effect. These observations are currently being fur ther investigated.
DNA of the cloned marker centromere has formed the basis for the construction of human artificial chro mosomes (HACs). An artificial chromosome is a defined structure that carries all the necessary fonctional elements for its long-term survival, replication, and segregation in a cell. These HACs are important since they not only provide direct proof for and permit the further dissection of the functional components of the marker centromere, they will provide a gene-delivery vehicle that has a number of distinct advantages over those that are presently used for gene therapy. A number of test constructs containing the marker centromere DNA have already been prepared and are being tested in mammalian cell cultures for their ability to remain stably as extrachromosomal entities. "Knocking Out' the Centronrtere Proteins. We are also studying the effect of "knocking out" a number of the known centromere-binding proteins in tissue culture and in transgenic mice. Since the centromere proteins are essential for proper chromosomal segregation, we hypothesise that mutation in these centromere-binding proteins is a major cause of chromosomal aneuploidy and/or cell cycle perturbation that will result in, amongst other manifestations, fetal loss during pregnancy. We have now produced null mutations (i.e. homozygous gene knockout) for two of these proteins. Interestingly, one of the proteins has demonstrated a lethal phenotype while the second protein appears to show no overt phenotypic abnormality in tissue culture.
Centromere Studies. A major effort of the laboratory has been in the study of a mitotically active centromere of a human chromosome 10-derived marker chromosome that is devoid of the usual large amount of repetitive a-satellite DNA. Chromosome walking {using FISH (fluorescence in situ hybridisation) and anti-centromere antibody staining} has now identified a series of YACs (yeast artificial chromosomes), PACs (PI-derived bacterial artificial chromosomes) and cosmid clones that span the centromere and has narrowed the functionally critical region from 200kb in 1995 to less than 80 kb. Extensive restriction mapping and partial sequence comparison of the marker centromere DNA with the corresponding region of the normal chromosome 10 where the neocentromere has appeared has demonstrated no significant difference between the two DNA, suggesting that the neocentromere has originated through the activation of a latent centromere DNA. The cloned marker centromere DNA region is currently being characterised in a number of different ways. DNA sequence confirms the lack of a-satellite DNA and reveals short stretches of some other unrelated tandem repeats. Using gel electrophoretic mobility shift assay, at leaist one of the sequenced repeats has been Page 11
t
We have recently received notification from Nature Genetics of the acceptance for publication of a paper on our work entitled “A functional neo-centromere formed through activation of a latent human centromere and consisting of non-alpha-satellite DNA"
Liz Earle analysing cells for centromere proteins
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TLeM uiFciocli Insttifiitte Iraco
Julian F.B. Mercer Sharon La Fontaine Daniel Strausak Andrew Grimes Stephen Firth Jenny Paynter Sharon Horton Loreta Ambrosini Michael Petris Michael Theophilos Paul Lockhart Karl Brand Michelle Howie Jim Camakaris
Group
Group Leader Postdoctoral Fellow Postdoctoral Fellow (from Nov. 1996) Research Officer Research Assistant Research Assistant Research Officer Ph.D. Scholar Ph.D. Scholar Helen M. Schutt Ph.D. Scholar Ph.D. Scholar Honours student Honours student (jointly with LaTrobe University) Senior Associate, University of Melbourne
the isolation of the gene affected in Menkes disease and this has led to further exciting studies which are beginning to unravel the complex processes which regulate the amount of copper in the body. We are studying the mutations which cause the disease in the families affected by Menkes disease, and we hope that this study may lead to better diagnosis and treatment. We are also investigating ways of correcting the genetic defect in Menkes disease by putting a normal gene into cells from Menkes patients. Over the next few years we intend to use the mouse models of Menkes and Wilson diseases to study the possibility of gene correction in whole animals. This process will enable us to study the normal function of the gene and may lead ultimately to gene therapy of Menkes and Wilson diseases.
Menkes Disease. The element copper is vital for life. Copper is required for the production of about seven enzymes, one of which is involved in producing energy. The amount that exists in each cell, although small, is critical. Julian Mercer and the Trace Element Group are studying how this essential trace element is carried around the body and within cells. Their particular interest is to determine the molecular basis of the genetic diseases such as Menkes disease and Wilson disease which disrupt this process of copper transport. Too much copper in the system can lead to cell damage. Wilson disease is a copper toxicosis condition which leads to liver failure or brain damage and death in the teenage years unless treated. Menkes disease is a genetic copper deficiency, resulting in babies who have severe brain damage, low body temperature, coarse hair and loose skin and joints. They die in early childhood. Over the last few years we have achieved a major breakthrough with Page 13
In collaboration with Jim Camakaris's laboratory (University of Melbourne), we have been investigating the intracellular location of the Menkes protein (MNK) in Chinese hamster ovary cells. This model cell system has led to a very important discovery which may show how cells regulate their copper status. Some of the key findings are that MNK is located primarily in the transGolgi network (TGN) of the cell, but is continuously recycling between the TGN and the plasma membrane. The amount of MNK found on the plasma membrane is increased when the cell is exposed to high copper and this presumably allows the cell to efflux excess copper. When copper levels in the cell fall, MNK returns to the TGN. A major paper describing this work has been published in EMBO Journal. Further investigation of the molecular basis of this copper^regulated MNK movement is important not only because it might be the basis of copper homeostasis at a cellular level, but also because it is a novel mechanism in cell biology. It is important to establish whether this process occurs in different cell types; i.e. Is it a general mechanism or is copper trafficking of MNK a phenomenon only found in copper resistant cells? Does this mechanism help us to understand the clinical defects in Menkes disease? We are also interested in determining where and when MNK is expressed during development, and to under stand the role of the closely related copper transporter, WND which is affected in Wilson disease. To understand the molecular basis of the copper induced movement of MNK we have constructed a cDNA encoding the complete protein sequence of the protein. This has proved to be a difficult task because this DNA sequence was unstable when we tried to propagate the plasmids in E. coli. By Page 14
construction of some novel vectors we have overcome the instability problems and this has allowed us to prepare plasmids for expression in mammalian cells. Initial experiments showed that the gene could be transfected into COS cells and expression of the Menkes gene from the construct was detected, however, immunocytochemistry of the cells was not very informative since there was a high background and the expressing cells in the population made so much MNK that the localisation could not be determined. In subsequent experiments we have established CHO cell lines which express differing levels of MNK from the construct. Preliminary data indicates that the protein is localised to the transGolgi, as in the copper resistant CHO cells. Most importantly, copper also induces a redistribution of the MNK, thus we have a system in place to determine which amino acid residues are responsible for the copper response. The MNK expressing cells are also copper resistant. In the next 12 months we hope to exploit this system to study the effect of mutations in recognised functional domains as well as to test the effect of mutations found in Menkes patients on the activity of MNK. We have identified the mutation responsible for the brindled mouse. This mouse mutant is the closest murine homologue of Menkes disease, and has been used for studies on copper therapy, so the nature of the mutation is of considerable interest. We found that MNK from the mutant has a deletion of two amino acids in a well conserved region of the molecule, but a region which has not been well charac' terised for its role in the activity of the protein. By using a MNK-specific antiserum on western blots, we showed that the brindled mutant has normal amounts of pro' tein in its tissues, but presumably this protein can have only residual copper transport activity. In collaboration with Don Newgreen's embryology group we used the antibody to determine the location of MNK in the kidney of the normal and mutant mouse. MNK was found in the proximal tubules of the kidney, but not in the glomerulus. The distal tubules were also stained and the protein appeared to be localised to the basolateral surface of these cells. These are the first observations of the location of MNK in tissues, and suggests that the intracellular location may differ in various cell types. MNK is thought to be involved in the uptake of copper from the urine, but the significance of the specific distribution in the distal tubules is unclear.
I
bling those found in the normal mammary gland and so can be used as a model system to study copper transport into milk. The requirement for copper is particularly high in young animals, but nothing is known about the process of secretion of copper into milk. Using the MNK antiserum. Dr Ackland has demonstrated MNK in the Golgi of the PMC 42 cells, and the intracellular distribution of MNK is also altered by copper treatment of the cells. These results support the CHO cell studies and suggest the Cu'induced movement is not restricted to copper resistant cells. It is likely that MNK is required for copper secretion into milk. Sharon Horton operating the atomic absorption spectrophotometer
We continue to analyse some of our Menkes patients to identify mutations. Initially work is focussed on one patient who has been treated successfully with copper and another patient with mild Menkes disease. Both these patients have normal amounts of MNK mRNA, which suggests that some MNK protein may be formed, which could explain the good response to treatment and the mild disease in the second patient. The sequence of the MNK cDNA from these patients has almost been completed and the results will be of clinical interest, perhaps leading to DNA tests for carriers.
Wilson Disease and Toxic Milk Mice. Animal models of Wilson disease are another area of ongoing investigation. The mouse models (toxic milk) accumulate copper to very high levels in the liver, in a similar manner to human patients with Wilson disease. We have found a point mutation in the Wilson gene homologue in the mutant mouse which causes a substitution of a valine for a methionine in the copper channel through the membrane. This result proves that the mouse is a true model of Wilson disease and therefore opens the way to more extensive use of this mouse for studies on treatment of this disease. We are also investigating the protective role of the small metahbinding proteins, metallothioneins (MTs) in Wilson disease. The excess copper in the mutant liver is bound to MTs and it has been suggested that variation in the ability to produce MTs in response to copper may explain some of the clinical variation in Wilson disease. We are investigating the role of MTs in the toxic milk mouse, by breeding the tx mutation onto a mouse line which cannot make MTs .This breeding is quite complex, but we are hopeful that some double mutants will soon be available for analysis.
We are also collaborating with Leigh Ackland of Deakin University to determine the localisation of MNK in a breast cancer cell line PMC42. This cell line is derived from a stem cell which can differentiate in culture to form ductal structures resem' Page 15
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lyiiuLFdlocli InstitTULte
G ene
Jeintifica^ion amd Expression Group
ii Henrik Dahl Steven Wilcox Wendy Hutchison Amelia Csborn Laraine Peters Sarah White
Group Leader Postdoctoral Fellow Scientific Cfficer Research Assistant PhD Scholar PhD Scholar
Finding the right genes, making mouse models and correcting mutations is the basis of our studies on disorders of mitochondrial energy deficiency. These are highly variable diseases which often affect brain and muscle function. Mitochondrial disorders are relatively common and are caused by mutations in a variety of genes. They can also contribute
Mitochondrial Energy Production. Energy generation is an essential and very complex cellular metabolic process involving many enzymes. It is therefore not surprising that mitochondrial diseases are relatively common, with a combined frequency of approximately 1 in 5000 births. They are caused by mutations in a number of genes, some of which are encoded on one of the 23 pairs of chromosomes present in the cell nucleus. Other genes are present on the abundant and maternally inherited mitochondrial DNA. The clinical presentation in affected patients can vary significantly, making mitochondrial diseases difficult to diagnose. Brain and muscle function are often affected, especially as is the case with the severe paediatric cases seen at the Royal Children's Hospital. Mitochondrial energy production decreases with age and this hais been shown to be a contributing factor in some late-onset diseases, including diabetes, Alzheimer disease and general dementia. Further understanding of diseases caused by mitochondrial DNA mutations requires an animal model. Collaborative studies on disorders of mitochondrial energy deficiency continue with David Thorburn's metabolic research group.
to late-onset disorders such as diabetes, Alzheimer and Parkinson disease and other dementia. Our research studies have focussed on: - the identification of nuclear genes causing mitochondrial dysfunction, '
generating a mouse model for diseases caused by defects in the
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mitochondrial DNA, correcting mutations in the mitochondrial DNA by introducing a normal gene into affected cells.
New projects have begun aimed at characterising the molecular defects and their cellular consequences in families with chondrodysplasia
Mitochondrial DNA and the ATPased Gene. While it has not yet been possible to engineer mutations into mitochondrial DNA, in collaboration with Jeff Mann, Beckman Institute, California, we have generated mice that should carry mitochondrial DNA from two different mouse strains. These mitochondrial DNAs can easily be distinguished and will initially be used to study the inheritance of mitochondrial DNA and especially the so-called "bottleneck”, a mechanism during oogenesis (egg development), where the proportion of a mutation, at least in humans, can change dramatically. We are also trying to correct the effect of defects in the mitochondrial DNA by synthesising and expressing a nuclear homologue of the mutated gene. Cur focus has been on the ATPased gene. A problem is that expression of the nuclear version of the gene appears to be lethal to cells. This has necessitated the use of special cloning vectors that allow highly controlled expression of the gene. It is also leading to the isolation and characterisation of control elements responsible for correct expression of nuclear encoded mitochondrial proteins.
punctata and Bethlem myopathy. We have also begun the search for a gene causing a form of inherited deafness which affects approximately 1 in 8000 children.
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Complex 1 Deficiency. As part of the mapping efforts of the group, we have been investigating a patient with respiratory chain Complex 1 deficiency. The patient has a chromosome 11:18 translocation and using FISH (fluorescent in situ hybridisation) analysis the chromosome breakpoints have been mapped to specific yeast artificial chromosomes (YACs) containing human DNA. Further research should provide us with the exact location of the breakpoint and allow us to investigate genes affected by the chromosome breakage. Page 18
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G ene Arylsulfatase E Gene. Mutations in the newly discovered arylsulfatase E gene on the X chromosome have been found to be present in individual patients with chondrodysplasia punctata, a group of bone dysplasias characterised by punctata changes around developing epiphyses seen radiologically in infants. Les Sheffield has a longstanding interest in this disorder, and we have collaborated on characterising molecular defects in Australian patients. Using single-stranded conformation polymorphism analysis and DNA sequencing we have screened the arylsulfatase C and E genes and identified the mutations in three Australian families. Although the families have different mutations they all cause amino acid substitutions in the arylsulfatase E gene.
Bethlem myopathy. Bethlem myopathy is a muscle disease characterised by slow progressive limb-girdle muscular atrophy and weakness, and contractures of multiple joints. Genetically it is a dominant disease which has been associated with mutations in the collagen 6A1 and A2 genes on chromosome 21q22.3 and the collagen 6A3 gene on chromosome 2q37. In collaboration with Mac Gardner and John Bateman's group in the Royal Children's Hospital we have been investigating the molecular defect and cellular consequences in a Victorian family with Bethlem myopathy. Using linkage analysis we mapped the defect in our family to the same region on chromosome 21 that contains the collagen 6 genes, and are now searching for the causative mutations in these genes.
Inherited Childhood Deafness. Inherited childhood deafness has an incidence of about 1/2000 births and autosomal recessive and dominant, X-linked and mitochondrial inheritance has been observed. Congenital isolated (nonsyndromic) autosomal recessive deafness (NSRD) accounts for approximately 50% of childhood prelingual deafness, and is a serious clinical and social problem. Several NSRD gene loci have been suggested. The first to be mapped was DFNBl, a locus on chromosome 13ql 1. We now know that this locus contributes to 50% of NSRD in the Caucasian population. In collaboration with Mac Gardner and Marion Maw we now believe that the DFNB1 gene is located on a YAC clone. This has been identified by FISH analysis. By island rescue PGR we have isolated several genes from this YAC and are presently analysing these candidate genes. Included in this analysis is a search for mutations by comparison of DNA gene sequences in unaffected people and people with deafness caused by a DFNB 1 gene defect. We also plan to use other techniques, such as exon trapping and expressed sequence tag analysis, to search for candidate genes in the relevant chromosome region. Henrik Dahl preparing amplified DNA
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Bob Williamson Kathy Williamson Tracy Evans-Whipp Kumaran Narayanan Louise Wangerek Damien Paris Stuart Beattie
Group leader Postdoctoral Fellow Postdoctoral Fellow PhD Scholar PhD Scholar Research Assistant Research Assistant
The Drea.m of Gene Therapy. For generations, those of us who help to look after the medical needs of young people with birth handicaps caused by a genetic mistake in the DNA code have had a dream. The dream is that one day we would be able to take a normal copy of a human gene and use it as a medicine, to provide the cellular function that cannot be performed because the child's DNA is mutated.
Before coming to Australia I had the privilege of heading the first liposome gene therapy trial for cystic fibrosis (CF). It was quite successful and proved that the treatment corrected the ion transport defect for a short time, and did not cause harm to our patient volunteers. This was enough to encourage people to continue to the next step - repeated gene transfers to the lungs of patients - and those trails are now under way in London and Edinburgh. Since I did not want to repeat the same experiments here in Melbourne, we decided to focus on other approaches - using lipids together with specific targeting molecules, with both DNA and RNA copies of genes for the central and peripheral nervous systems. Our immediate targets are Friedreich's ataxia (for which we have generous support from the Friedreich's ataxia groups in Victoria and Queensland, from APEX Elmore, and from private benefactors in New South Wales), dementia in Down syndrome, and epilepsy. The complexity of the central and peripheral nervous systems makes work in this Page 20
area particularly challenging. However, these studies are worthy of our attention since degeneration of the nervous system is a feature of many of the most severe inherited diseases. Friedreich's ataixia is a particularly relevant model for this class of disease. Patients suffering from this form of neurodegeneration lose control of limb coordination (ataxia) and are commonly wheelchair bound by the third decade of life. The gene for Friedreich's ataxia was identified in 1996, and although the role the gene plays within the cell is still not known, mutations present in patients have been characterised. The predominant mutation is an intronic expansion of a GAA trinucleotide repeat, which is present in both gene copies in affected individuals and is predicted to cause lack of the associated protein. Damien Paris and Martin Delatycki have studied the pattern of mutations present in Friedreich's ataxia patients from Victoria, Queensland and New South Wales. Their findings demonstrate concordance between clinical and molecular diagnoses. For example, the length of the trinucleotide repeat expansion appears to correlate with severity of disease; patients with shorter expansion lengths tend to have a milder disease presentation. Furthermore, we have shown that expansion length can increase or decrease during parent to offspring transmission, as well as within the cells of an individual. Indeed, Friedreich's ataixia is the only known disease to display frequent shortening of a trinucleotide repeat expansion (this feature is observed occasionally in other trinucleotide repeat diseases such as myotonic dystrophy and X^linked mental retardation). Kathy Williamson's interests focus on the structure of the Friedreich's ataxia gene, particularly the promoter region. Analysis of this region will aid identification of the sequence required for correct expression of the gene, and hence this sequence can then be incorporated in gene therapy constructs. Luciferase reporter assays, involving transient transfection of appropriate cell lines, are being employed in this area of research. The expression pattern of the Friedreich's ataxia gene, in humans and mice, is an additional area of interest. The best mode of transfer of material to neuronal cells is an integral part of the gene therapy programme. We are developing several different strategies for introducing genes into cells of the central and peripheral nervous systems. Liposomes have been relatively successful in mediating gene transfer to the cells Louise Wangerek examining transfected
of the lung airways of patients with CF but their ability to transfer genes to nerve cells has not been fully aissessed. Louise Wangerek in the group has demonstrated that liposomes can indeed shuttle genes into cells both in culture and in the mouse brain. We are also investigating the use of viruses which naturally infect neuronal cells to deliver therapeutic genes. Tracy Evans-Whipp, who joined us from London in November, is developing vectors based on the Herpes Simplex virus which will carry genes into neuronal cells but do not encode any of the potentially harmful functions of the virus. In parallel, Kumaran Narayanan is studying whether RNA can be used to transfect neurons in a model for Menkes disease, since this alternative method may be particularly suitable for a non-dividing tissue such as the brain or spinal cord which has very low ribonuclease activity. In Menkes disease, copper does not pass properly across the blood-brain barrier, leading to severe neurological disease and mental retardation. Even though this disease is relatively rare, gene delivery via the cerebrospinal fluid may enable correction of the copper transport failure. This research is being carried out in conjunction with the Trace Element group headed by Julian Mercer. We are not, however, confining our research to the nervous system. We are continuing work on cystic fibrosis with Brandon Wainwright, visiting from Brisbane for a year during 1997. We are also fortunate to have Panos loannou with us. Panos heads the Molecular Genetics unit in Cyprus where the prevalence of (3-thalassaemia is a major burden on the health system. P thalassaemia may pose fewer problems for gene therapy than some other diseases. By taking a blood sample it is possible to obtain cells easily from patients for studies of gene transfer into human “stem cells", which can divide many times in the body. Our experiences with thalassaemia will not only help people in Australia who have this disorder but also those in other countries, particularly those in the Mediterranean area. These studies may also provide clues on how to treat leukaemia (which similarly affects the precursors of blood cells) using genes more effectively. What, you may ask, is the time scale for turning these exciting ideas into therapy? In the case of ataxia there are three major difficulties. The first is the sheer complexity of the brain itself and the need to understand which neurons are involved in any particular disorder. The second is the need to find new ways to introduce genes into neurons without causing inflammatory or immune reactions. The third will be to ensure complete safety for the patients. We are pleased that we have started on the road to gene therapy, but the road may be a long one!
cells
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IIFC oc. 1
M^efatolic R eseaFcn
David Thorburn Denise Kirby Kirsty Reed,
Unit Head; Research Officer/PhD Scholar; Honours student.
enzyme complexes of the respiratory chain, complex IV and complex 1. This involves both somatic cell genetics (in conjunction with Kerry Fowler and Marjorie Crawford) and molecular biology (in conjunction with Henrik Dahfs group).
Complex IV (cytochrome c oxidase). Approximately 1 in 5000 children will suffer from disorders of energy production which can result in developmental delay, movement disorders, liver and heart failure and muscle weakness. This can be a result of severe respiratory chain dysfunction, and the inheritance can be autosomal dominant, autosomal recessive, Xdinked or maternal. Our main research efforts are directed at identifying the nuclear gene defects that cause respiratory chain dysfunction and clarifying the genetics of mtDNA inheritance. The mitochondrial respiratory chain is responsible for the bulk of energy production from all cellular fuels. It is composed of approximately 80 different proteins organised into five enzyme complexes in the inner membrane of mitochondria. We now know that over 150 genes are involved in encoding the different proteins of the respiratory chain and in coordinating the import and assembly of these proteins within mitochondria. Some of these genes are encoded by mitochondrial DNA (mtDNA), present in thousands of copies per cell, but most are nuclear genes.
We have constructed "cybrids” from patient cell lines with complex IV (cytochrome c oxidase) deficiency. Cybrids are cytoplasmic hybrids, containing mtDNA from the patient but a nuclear genome from a normal person. Results of cybrid studies and mutation analysis of the 10 nuclear gene subunits of complex IV indicate that the patients have a nuclear-encoded defect but apparently not in the subunit genes. Now we are trying to identify other candidate genes for complex IV deficiency. We are studying a subgroup of patients from consanguineous Lebanese families, suggesting that they may have inherited the same mutation from a common ancestor (i.e. a founder effect). Complementation analysis (see Figure) of patient cell lines shows that cell lines from four Lebanese families do not complement each other, but can be complemented by a cell line from an Anglo-Australian complex IV patient. This implies that the Lebanese families have mutations in the same gene, and that the Anglo-Australian patient has a mutation in a different gene. We are currently performing homozygosity mapping of these families (in collaboration with Simon Foote, Walter &- Eliza Hall Institute) to identify a genomic locus for complex,IV deficiency.
COX'deficient cells from patient A
selection
COX'deficient cells from patient B
Heterokaryon, with nuclear genomes from both patients
Somatic cell complementation analysis of fibroblasts from patients with respiratory chain complex IV (COX) deficiency. If COX activity is restored in heterokaryons, then the patients are in different complementation groups (ie. different gene loci). If not, the patients are in the same complementation group and pre sumed to have mutations in the same gene.
Complex I. Cell Lines and "Cybrids". Over the past two years our metabolic research group has successfully characterised cell lines from patients with respiratory chain defects. We have done this by determining the precise enzyme defect and then assessing the functional effects of such defects on parameters such as ATP synthesis and growth on poor substrates. We are now able to focus on two groups of patients with defects in two of the Page 23
Our more recent research shows that complex I deficiency is the most common type of respiratory chain defect in children, but its extreme complexity (more than 40 subunits, including seven encoded by mtDNA) has meant we know little of its genetic basis. We have commenced cybrid and complementation analyses of our patient cell lines to determine how many have nuclear defects and how many different genes cause complex I deficiency. Preliminary results suggest most are likely to have nuclear defects and we have begun studying a candidate gene for complex I deficiency.
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TkeM nirdocli I ns tilt mite Hi/nn DFyology
Cj'Foiip
(I Don Newgreen Susan Bevan Joseph Minichiello Richard Kerr Catherine Hearn Sonja Jackson
Group Leader Postdoctoral Fellow (till Oct. 1996) Research Assistant PhD Scholar PhD Scholar Honours Student
!J
mtDNA Inheritance.
The Embryology Group studies the very early development of an
Genetic counselling of families with mtDNA mutations poses major problems as it has not been possible to predict accurate recurrence risks, and there has been dis' agreement about whether prenatal diagnosis should be attempted. We have quanti' fied the mutant load in tissues from 13 families we identified with mutations at nt.8993, and surveyed the data on 32 other published families. This has allowed us to generate empirical recurrence risk data and convinced us that prenatal diagnosis can be offered to some families provided they are carefully counselled about the limi' tations of interpreting mutant loads measured in chorionic villus.
embryo at the time when cells are differentiating and organs are beginning to form. Fertilised quail eggs provide convenient research material since, at these early stages, basic developmental processes are similar in birds and mammals (including humans). Already in development, the location and differentiation of the different body tissues has begun, and tissues, organs (eg liver, heart) and systems (eg reproductive, nervous, circulatory) are taking form. Any developmental errors which occur at this time will subsequently lead to major physical malformations such as spina bifida, cleft palate and heart malformation. The cells of the neural crest seem to be key players in early development. The neural crest cells come from the neural tube, which will go on to form the brain and the spinal cord. These cells perform their role by physically moving (or migrating) from one site to another to construct new tissues of many different types. For example, they form and sculpt the shape of the face. They are also responsible for normal development of the heart. When they divide the single artery leaving the early heart into two separate arteries, one will end up taking blood through the lungs and the other to the rest of the body. Abnormalities in the migration of neural crest cells is all that is required
Denise Kirby assaying respiratory chain enzymes at the spectrophotometer
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for such common developmental problems as facial clefting (cleft lip and Page 26
begin to move. Cell biological and immunological studies in our laboratory suggest that for cell movement to occur, this protein acts as a link between adhesion (traction) molecules at the cell surface and cytoskele^ tal (motor) molecules within the cell.
palate) and cardiac malformations to arise. This migration is influenced by the molecules present in the developing body. If due to a genetic defect, the correct molecules required are not available to the cells, malformations can occur. By looking at molecular control of neural crest cell development, as well as using what we learn from the birth defects which arise when this is abnormal, we can learn much more about the basic developmental process. The Onset of Cell Movement. A dramatic event in early development is the sudden transformation of neural crest cells from a group of stationary cells, held together by intercellular "glue”, to moving individual cells which invade surrounding tissues. Events like this are the basis of many changes in development, and also occur in uncontrolled form in the malignant transformation of carcinomais. We have made cell movement a focus of research. Our spatiotemporally specific cDNA libraries pointed to changes in growth factors of the TGF'b superfamily and their receptors at the time of neural crest cell movement, suggesting that they could switch on this process. We devised a tissue culture model which mimics the onset of cell migration and applied one of these factors which is normally present and a related factor which is not found at the time. Both factors stimulated the onset of cell migration with the appropriate factor being vastly more effective. All growth factors operate via cascades of intracellular signals beginning at cell surface receptors and ending at effector molecules in the cytoplasm and cell nucleus. Using drugs which modulate signal molecule function we have shown that downstream of the receptor lies a signal transduction enzyme of protein kinase^C type, probably an iota or zeta isoform. Protein kinases alter the function of downstream protein molecules by reversibly adding a phosphate group. We have used radioactive phosphate and biochemical analysis to show that the amount of phosphate on many proteins alters, Richard Kerr microdissecting a but one molecule alters before all the others as cells
Control of Cell Movement. The migration of neural crest cells depends on reversible adhesion to extracellular matrix molecules and to cells. Recent experiments with certain embryonic matrix proteoglycans isolated in our laboratory show that these inhibit adhesion and migration. The type of proteoglycan is not important when it is attached to a surface of adhesive matrix molecules on which cells move: this is probably due simply to the very large proteoglycan covering the smaller adhesive molecules. Surprisingly, proteoglycans are also inhibitory in solution, but this is specific for the type of proteoglycan. Even more surprisingly, specific proteoglycans also destabilise specific celftO'cell adhesions. This suggests in particular that these molecules could play a role in the "ungluing” of cells at the start of cell movement. Of more general importance, however, this discovery places the molecular control of loss of cell-tO'Cell adhesion on an equal footing to the much bet' ter investigated making and maintaining of adhesion. Hirschsprung Disease: a Cel! Movement Disorder.
!
Neural crest cells colonise the intestine with nerve cell precursors by an orahto^anal migration. A defect in neural crest cell migration has long been proposed to underlie Hirschsprung disease, where the nervous system fails to form in the colon. Molecular genetic studies have identified defects in two growth factor/receptor systems (GDNF/ret and ET3/ednrB) as causing some forms of this condition. However, the link between genotype and phenotype is still obscure. We have succeeded in the difficult taisk of isolating the neural crest cells that produce the nervous system in the intestine and tested them in tissue culture with GONE and ET3. Both these factors encourage initial neural crest cell division but later GONE promotes differentiation of nerve cells which can no longer divide, while ET3 opposes this. In our tissue culture model system, reduced migration of the cell population is caused by lowering the number of cells. We also showed that there is a tight spatiotemporal relationship between the migration of neural crest cells in the intestine and the growth of that organ. This led us to propose that a slight delay in migration consequent to a subnormal number of migrating cells can lead to the migration end-point growing away from the migrating cells, with consequent failure to completely colonise the colon, typical of Hirschsprung disease.
Future Directions. These projects will be extended by further molecular genetic studies using the cDNA libraries on the growth factor requirements of the onset of migration, which are clearly more complex than the few TGF-
quail neural tube
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instifiite
M OMse WloJel Unit b family members and their receptors which we have identified so far. Likewise the molecular genetics of Hirschsprung disease is clearly incomplete since mutations in genes for the identified growth factors and receptors account for only a minority of cases. This will be pursued using spatiotemporal cDNA libraries, in collaboration with colleagues overseas, and in our laboratory will examine the roles of other neur^ al growth factors. The chain of events leading to the onset of cell migration will be followed up by exploiting an immortalised neural precursor cell line to overcome the difficulties of small sample size which are imposed by normal embryonic tissue sources. This resource will aid the isolation and identification of the phosphorylated protein which we have shown is central to the start of cell migration. The controlling role of certain proteoglycans is now clear, but it is not clear how this control is exert' ed. Our evidence points to signal transduction steps set in motion on contact of the proteoglycan with the cell surface, and these will be investigated using techniques already used in studying the onset of cell migration. These studies are illuminating the web of interactions at the genetic and epigenetic levels that enable the daunting complexities of development to proceed.
Kerry Fowler Sophie Gazeas Robyn Breslin Joanna Hill
Senior Research Officer Senior Technical Officer Technical Assistant (Part Time) Research Assistant
The usually humble mouse is of great significance at the Murdoch Institute. Under the leadership of Kerry Fowler, the Mouse Model Unit provides mice which are accurate models for human disease. By using the process of transgenesis, new genetic material is introduced into the mice creating precise models. This assists us in our research and enables the development and testing of therapy strategies which, with adherence to strict criteria, can then be trialed in humans. Over the past two years there has been a rapid increase in demand for transgenic technology from within the Murdoch Institute as well by the Royal Children's HospitsJ Research Foundation (RCHRF). Our new laboratory is now located at the Murdoch Institute, and an extension of the mouse breeding facility at the Royal Children's Flospital is planned in liaison with the RCFIRF and Melbourne University's Department of Paediatrics.
Helping Chromosome, Copper and Mitochondria Research. One of our 1996 successes has been to produce mice which are unable to produce the centromere'binding proteins B and C. Normal epithelial cells like these skin cells in panel A are held together by adhesion molecules (labelled in green) and the cell structure is maintained by a cellular skeleton (labelled in red). In panel B, the cytoskeleton has been disassembled using drugs to investigate how adhesion molecules are anchored to it. The nuclei of these cells are labelled blue.
These mice are being carefully studied by Andy Choo’s chromosome research group to look for chromosomal abnormalities associated with cell division as well as sperm and egg production.
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1 issue Culfuire
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Julian Mercer's trace element group also uses the unit to generate mice which have mutations in more than one gene. Breeding lines which have Wilson disease, the result of a copper overload, as well as mutations in metallothionein 1 and II genes, are also being studied. With Henrik Dahl's mitochondrial energy group and, in collaboration with Jeff Mann, at California's Beckman Institute we continue to develop transgenic mouse models for mitochondrial disease. Mitochondria inheritance is through the mother and, given the difficulty of introducing mutations into the mitochondria genome, this project has proven to be particularly challenging.
Gene Therapy. Plans are also under way with Bob Williamson's gene therapy group to generate mouse models for studying neurogenetic disorders such as ataxia.
External Collaborations. Expert transgenic facilities are scarce and we are collaborating with other group to develop models for human disease. Kennedy disease mouse model (with Garry Warne's Endocrinology group at the Royal Children's Hospital Research Foundation). Connective tissue disease mouse models (with John Bateman's Orthopaedic Molecular Biology Research Unit, Department of Paediatrics and the Royal Children's Hospital Research Foundation and Marie Dziadek's Anatomy and Cell Biology group at the University of Melbourne). Genes involved with male sterility (with John Hutson's Surgical Research Unit at the Royal Children's Hospital Research Foundation).
Kerry Fowler making glass micro-injection pipettes
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Marjorie Crawford Tiffany Symes Alison Blake
Scientific Officer Research Assistant Technical Assistant
The Tissue Culture Laboratory provides cell cultures which are vital ingredients for the ongoing research throughout the Institute. These cultures are used by the various scientific groups to look for mutations, chromosome abnormalities or to analyse enzyme levels which might indicate a metabolic disease. The laboratory also main tains a bank of cells in liquid nitrogen for future use and reference. New projects over the past year requiring a large component of cell culture have vastly increased our workload and consequently the demand for cell culture media (see graph). This has put a great strain on our facilities and we have appreciated the conversion of the nearby instrument room into a second laboratory. To keep up with the demand we have also been training laboratory staff in the procedures of tissue culture.
. fr '?Wi
mm
w
swii
Litres of media made per year
litres 1400.00 ^
1200.00 1000.00 -
800.00 -
600.00 400.00 200.00 -
0.00 1991
1992
I
I
1993
1994
year
1 1995
1996
Statistics of our activities also show a " . „ -I significant increase in the number of cell . aS lines received from other laboratories. 3 This increased from 68 in 1995 to 90 in 3 1996. Tiffany has recently completed i her Bachelor of Science at RMIT S graduating with distinction.
Tiffany Symes with cell samples in the warm room
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M'ii]r(iloc]li Institmte
HL^pidemniology
If
Jane Halliday Les Sheffield Carole Webley Ron Batagol Liz Taylor
Epidemiologist Medical Geneticist Research Assistant Research Assistant 5th year medical student
Maternal Serum Screening Program. The Epidemiology Unit has been a part of the steering committee working on the establishment of this program by the VCGS, and will be monitoring the outcome of pregnancies to determine the effectiveness of the screening program in terms of detection rate of Down syndrome, and the use of genetic counselling services.
Pregnancy outcome of cases where prenatal diagnosis determined an abnormality in the fetus.
Epidemiology is the branch of medical research which involves the study of the distribution and determinants of disease and health in populations. Epidemiological information is very useful for health planning, and forms the basis ofprogrammes designed to control or reduce health problems in the community. The Epidemiology Unit at the Murdoch is involved in many aspects of research into birth defects which are described below. The Unit has strong links with other health agencies, research groups and the Perinatal Data Collection Unit, Victorian Department of Human Services (headed by Jane Halliday), providing unique opportunities for collaborative research.
Carole Webley has been investigating how often parents choose to continue pregnancies where the fetus has been diagnosed with an abnormality and results will be available in 1997. Pregnancy outcome information has been obtained for over 300 fetuses with chromosome abnormalities ranging from major disorders such as Down syndrome to more minor conditions.
Mosaic karyotype. Another project instigated this year is a long-term follow-up of children diagnosed in utero with a mosaic karyotype, that is, some cells with normal chromosomes and other cells with abnormal chromosomes. The effects of these abnormalities are largely unknown. Liz Taylor, a 5th year medical student from the UK, worked with us to see how many liveborn babies there were in this group in the three year period 1990^1992, who would now range in age from 4-6 years. Ninety seven cases were identified in these years and we now know of another group of approximately 100 babies born between 1986 - 1989, who may also be available for follow-up. We are designing a case/control study and will be collaborating with experts to assess aspects of child health and development.
Prenatal Diagnosis With the increasing frequency of prenatal diagnosis, it is of vital importance to closely monitor amd atssess the services which are available and their value to the community. The Epidemiology Unit collates and analyses all the data on amniocentesis and chorion villus sampling for the state of Victoria. Our 1994-1995 combined prenatal diagnosis report was completed and published in October, 1996. In 1994-95 about 7% (4,500) of all pregnant women in Victoria had either amniocenteses or chorion villus sampling. Testing is recommended for women over the age of 37 years, and the majority of older women had one of the tests. We found that nearly 50% of all fetuses with Down syndrome were detected by the prenatal diagnostic service in 1994 but it should be noted that due to subsequent natural miscarriage and fetal death, not all of these pregnancies would have resulted in a live birth. Page 33
}
Predictive Genetic Testing. The Epidemiology Unit is involved in evaluating clinical services in other areas such as predictive genetic testing for cancer - an area which is changing very rapidly as more cancer-predisposing genes are identified. We are participating in an important international collaborative study of the psychological impact of predictive genetic testing for familial adenomatous polyposis (FAP), a rare bowel cancer. Presently it is unknown how adults and children will react to the results of DNA tests revealing a genetic risk of cancer. To ascertain the psychological effects of this programme, and to assist in future planning of services, participants are given a series of questionnaires to complete before and after testing. Forty three questionnaires were completed in Victoria in 1996 and data collection will continue in 1997. Analysis is being done on both a local and international basis.
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if'l
VicfoFian Climcai Genetics S ervices
-DP^A Diagnostic Latoratory
Drugs in Pregnancy.
MURDOCH INSTITUTE
Les Sheffield has continued to lead this study. Women have been recruited into the study from two obstetric drug information centres, and interviewed by genetic counsellors (Susan Clement and Clara Gaff) about their medication history. Carole Webley has analysed our data from a previous study and has designed a method to construct a profile of medication use for each pregnancy. The analysis has allowed for a check on whether babies with birth defects born at the Royal Women's Hospital were notified to the Victorian Perinatal Data Collection unit. All 23 babies found with a major birth defect in the study had been notified to the Perinatal Data Collection Unit. Ron Batagol has been a member of this research team for many years but left us at the end of the year to return to pharmacy practice. We thank him for his participation and enthusiasm.
Sue Forrest Ivan Biros Jan Brasch Jenny Douglais Karina Forshaw Steven Nasioulas Janet Shaw Tom Milovac Iswari Setianingsih Jan Fullerton
Scientist'in'charge Scientific Officer Medical Scientist Medical Scientist Medical Scientist Medical Scientist Medical Scientist Technical Assistant PhD Scholar Honours Student
MONASH MEDICAL CENTRE
Jean Hendy Andrea Twomey Kathy Garafolo
Medical Scientist Medical Scientist Trainee Scientist
Jane Halliday and Carole Webley
Genetic testing using DNA technology is welhestablished for a number of diseases including hereditary colon cancers, haemophilia, muscular dystrophy and Huntington disease. The DNA Diagnostic Laboratories of the Victorian Clinical Genetics Service provides quality DNA diagnostic testing for Victoria and Tasmania, and new advances and tests are com tinually being developed e.g. for hereditary nompolyposis colon cancer (HNPCC). Prenatal diagnosis for prevention of disease requires accurate DNA testing. Identification of diseasC'Causing mutations is necessary before gene therapy can be offered. For all these reasons we strive to provide a quality genetic testing service, with a progressive research com ponent. During the past year our staff numbers have increased to cope with the growing number of test requests and our laboratories have been involved in a number of research and development projects. In addition to routine testing the diagnostic staff have worked to increase the number of different techniques available in the laboratory to identify genetic changes. This
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will enable us to offer direct testing for a wider range of inherited disorders. To cope with the ever-increasing number of gene tests requested for very rare conditions we have instigated a service for clinicians whereby we liaise with overseas laboratories to send prepared DNA away for testing (Tom Milovac). We have now identified the family-specific DNA change in 33 Victorian familial adenomatous polyposis colon cancer (FAP) families. Our first B.Sc. Honours candidate (Jan Fullerton) contributed to the search for these mutations. The demand for presymptomatic testing from FAP families is increasing. We have an additional member of staff (Jenny Douglas) to help with this testing and to extend our work on colon cancer to another group of patients with hereditary non-polyposis coli cancer (HNPCC). We are investigating different methodologies for HNPCC testing because the mutations found most frequently in these families are different from those in FAP families.
At the Murdoch Institute laboratory we also provided valuable experience for three students during the summer. During the year the thalassaemia DNA service (Jean Hendy) from the Royal Women’s Hospital moved to our Monash Medical Centre site bringing with it a large patient population. The challenge for the Monash laboratory this year has been to rationalise protocols for thalassaemia DNA testing and to provide testing for thalassaemia families from the different ethnic groups each of which carries its own range of mutations.
We continue to look to new developments in DNA diagnostics. Currently we have a clinical genetic fel low (Karen Dunn) investigating "Genetic Predisposition to Atopic Eczema”. This project reflects the thrust in genetic research to understand inherited tendencies of individuals to respond adversely to particular environmental factors. Prediction of such predispositions by genetic testing may enable prevention of some chronic and debilitating illnesses in the future.
Our involvement in testing for neuromuscular and neurological degenerative diseases has increased. We now test routinely for spinal muscular atrophy using a direct test as well as linkage. Trinucleotide repeat expansion was recently described as the causative mutation of Friedreich’s ataxia. This is a degenerative disease in which symptoms begin to appear around puberty. Based on research done in the diagnostic laboratory (Damien Paris) we should soon be able to offer a simple and accurate test to assist diagnosis and to detect carriers of this disease. This year, together with routine carrier testing and prenatal diagnosis for cystic fibrosis, we have been characterising the mutations in the Victorian population of affected cystic fibrosis patients. This project is designed to give a better understand ing of the range of CF mutations in our community and also to investigate variations in disease presentation and outcome which may be related to particular mutations. We need a thorough understanding of these aspects of a disease before we can offer gene therapy. Our PhD student, Iswari Setianingsih (Ning) from Indonesia has been characterising the mutations in p thalassaemia major and intermedia patients from her country. She is using the knowledge gained to set up DNA diagnostic services in the Eijkman Institute for Molecular Biology in Jakarta. Page 37
i"
Steven Nasioulas loading an acrylamide gel
Page 38 ■
Victorian Clinical Genetics S ervices
I M^etaloli1C laboratory
IV David Thorburn Ivan Francis Denise Kirby Erin Oldaker
Scientist-in-charge, Enzymology; Scientist-in-charge, Metabolism; Research Officer; Research Assistant.
The Metabolic Laboratory provides a diagnostic service for children suspected of having a metabolic disorder and also monitors affected children. These Inborn Errors of Metabolism are inherited disorders which affect the breakdown and refuse of fat, protein and sugars. Many of these disorders are severely disabling, but in some cases affected children can lead a normal life if their condition is recognised early. Erin Oldaker operating the amino acid analyser
Quantitative amino acid analysis is performed on plasma, urine or cerebrospinal fluid samples from any Victorian child suspected of a disorder of amino acid metabolism. Diagnosed children are usually put on special diets or given medication, for example to prevent the accumulation of natural compounds that may otherwise build up to very high (toxic) levels. We currently monitor over 100 such children from around Victoria to ensure their "metabolic control" is adequate to prevent long-term disability.
We act as the Australasian referral centre for diagnostic testing to measure enzymes in patients suspected of disorders of energy generation. All our organs, but particularly the brain, heart, muscle and liver, require energy to perform normally, and so any or all of these organs can be affected when our cellular power plants (mitochondria) do not work properly. The energetic capacity of patient muscle, liver or heart biopsies is measured by assay of the enzyme complexes of the mitochondrial respiratory chain. Interpretation of patient results is complicated by the secondary effects of "sick” tissues on the enzymes and it can be difficult to distinguish real genetic defects from secondary loss of enzyme activity. We have therefore studied Page 39
tissues available from patients with sick tissues due to other known causes, and used this data to establish more realistic reference ranges. The respiratory chain enzymes can also be assessed in patient cell lines (usually skin fibroblasts) together with other enzymes involved in energy generation such as pyruvate dehydrogenase, carboxylases and enzymes of the tricarboxylic acid cycle. Patient cell lines are also assessed by functional tests such as their ability to grow on poor substrates (galactose instead of glucose) or their maximal rate of ATP synthesis. As well as aiding diagnosis of these children, the cell lines form an important resource for research into the basic genetic mechanisms causing these disorders (see Metabolic Research section). New patients with inborn errors of metabolism diagnosed this year include 29 respi ratory chain defects, 10 (homozygous) cystinuria, 8 phenylketonuria, 2 pyruvate dehydrogenase defects plus isolated cases of several other inborn errors, including the first Australian patient diagnosed with fumarase deficiency.
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VicttoFian Clmicai Genetics S ervices
NewtoFii S creenmg
Ivan Francis Nick Tzanakos Len Bonacquisto Nella Napolitano Mona El'Masri
^(SL .50]P'
Scientist'in-charge; Medical Scientist; Medical Scientist; Trainee Medical Scientist; Trainee Medical Scientist;
The Newborn Screening Laboratory (NSL) has been in operation since 1966. It began by offering tests for phenylketonuria (PKU) using dried blood spots collected by heehprick from newborn babies before discharge from hospital. By 1972, every newborn in Victoria was covered by the PKU test or "Guthrie Test". In 1977, a test for congenital hypothyroidism (CH) was added; and in 1989 the NSL started screening for cystic fibrosis (CF). Victoria currently has about 65000 births per year; all are tested for these three serious diseases to ensure an early diagnosis, effective treatment and best possible outcome.
Phenylketonuria (PKU) is an inherited disorder of amino acid metabolism and occurs in 1/13000 births. When both parents are carriers of a PKU mutation, there is a 1 in 4 chance that their baby will have PKU. About 5 babies with PKU are born each year in Victoria. The baby must be placed on a diet low in the 'essential' amino acid phenylalanine, in order to avoid severe mental handicap. Before screening and dietary treatment was available, many of these infants spent their lives in institutional care; now they can look forward to full, productive lives. Congenital hypothyroidism (CH) is a predominately non-inherited disorder of the thyroid gland. It is caused by a deficiency of thyroid hormone, which is essential for normal skeletal and neurological development during the early stages of growth. Treatment by thyroid hormone replacement (one tablet per day for life) must be started at birth to prevent stunted growth and mental impairment (cretinism). In Victoria, CH occurs in 1/3100 births (20 per year). Page 41
Nella Napolitano sorting the daily delivery of Guthrie blood-spot cards.
Cystic Fibrosis (CF) is the most common life-limiting inherited disorder among Caucasians, occurring in 1/3200 births (20 per year) in Victoria. Newborn screening provides early diagnosis, ensuring appropriate therapy is available before debilitating malnourishment and respiratory disease has taken hold. The NSL is a part of the integrated services provided by the VCGS, and provides an important resource for ongoing genetic research within the Murdoch Institute. It is anticipated that advances in genetic knowledge will provide therapies for genetic disorders which are currently considered unbeatable. We will ensure that new developments are put into practice as they become available, thereby allowing early intervention in these disorders, thus contributing to the health and well-being of future generations.
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Victorian Clinical Genetics S ervices
Cytogenetics
^aoor.
Cytogeneticist in Charge Senior Cytogeneticist
Howard Slater Vida Petrovic
Scientists
Selena Bourke Trent Burgess Melissa Curtis Sue Dale Julie Davies Wali Drummond David Francis Julie Garratt Louise Hills Sarah Nouri Ralph Oertel Anne Robertson Marie Thorpe Tracy Vankuyk Cathryn Vaux Lucille Voullaire Lorna Webber (from Nov.'96)
e .
,^
Technicians
Ian Brooks Fernando Garcia Bo Jezierski
Lynda Phillips
The VCGS Cytogenetics Laboratory offers a comprehensive service in routine chromosome testing to hospital and private clinicians throughout Victoria and Tasmania. This includes detection of congenital and acquired (cancer) chromosome abnormalities. The laboratory also performs DNA testing for some syndromes formerly diagnosed using chromosome analysis such as the Fragile X, Prader-Willi and Angelman syndromes. The laboratory has special expertise in molecular Page 43
cytogenetics which combines tmditiona.1 cytogenetics with molecular biology to aid in the diagnosis of several microdeletion syndromes such as the Vehcardiofacial syndrome and Williams syndrome and also in the assessment ofpaediatric leukaemia. Some 4,700 tests in total were performed in 1996, a slight increase over the previous year. Changes and Advances. After one year in our new laboratories we are now very much at home beside our VCGS and Murdoch Institute colleagues. The benefits of working alongside other VCGS genetic pathology scientists, clinicians and the Murdoch Institute researchers is very obvious in terms of access to facilities and support and sharing in the academic environment of the Institute. This year, the laboratory made a major capital outlay in the purchaise of a Cytovision image analysis sys' tern. We threw caution aside and decided that the time had come for our laboratory to benefit from the latest computerised imaging technology. The particularly demanding and time-consuming routine paedi atric bone marrow work would be facilitated by "Cytovision" and we also needed an improved fluores cence system for our multicolour FISH tests. We are also investigating the potential of a new technique called Comparative Genomic Hybridisation(CGH) which may be very useful in situations where limited numbers of cells are available for analysis. A major development this year has been the introduction of prenatal chromosome testing. Two years ago the laboratory added chromosome testing of leukaemia patients to • our specialisation in paediatric bloods, and now the introduction of prenatal testing completes our involvement in what is recognised as the whole range of cytogenetic testing procedures. Prenatal is/-''"'’ karyotyping complements the in-house maternal serum screening ■ ff' programme for Down syndrome and along with the VCGS counselling services, offers a comprehensive screening service in this area.
3
Rare and Interesting Cases Research and publications continue to capitalise on interesting cases or groups of cases which were recognised within the diagnostic work of the laboratory. Howard Slater with the Cytovision image analysis equipment
Page 44
if: f.'
VicfoFiam Clinica Geneltics S CFTices !
ill! I iii' '
Generic C oanselling and. CanceF A patient with a very rare, biphenotypic lymphoma and a tumour associated with chromosomes 8 and 13 was investigated at some length in collaboration with Deon Venter and colleagues in the Department of Pathology at the Royal Children’s Hospital.. Initially, an involvement of the retinoblastoma tumour suppressor gene was suspected but this was shown not to be the case. Further collaborative studies are now in progress to identify the DNA sequences disrupted by this translocation. We have published one of the first examples of prenatal diagnosis of Prader Willi syndrome in a collaboration with the Royal Women’s Hospital cytogenetics laboratory. This work threw up two unexpected findings of scientific interest. Firstly, we showed there was unimaternal disomy for chromosome 15 and that “rescue ” had taken place from a trisomy 15 conception. What was surprising was that residual trisomy 15 cells were present which raises questions as to the timing of rescue and the clinical significance of “residual trisomy’’. Secondly, we discovered that the DNA locus for Prader-'Willi syndrome in one of our controls was not methylated which indicated that the widely held notion of complete methylation in amniocytes is wrong. We have published a series of Fragile X reverse mutations in an ongoing collaboration with Danuta Loesch of Latrobe University. Normally large Fragile X mutations are methylated, a process which inactivates the gene. We discovered a rare example of a large mutation in a high functioning male which had remained unmethylated.
The Future. There is a constant pressure for our laboratories to find clinical applications for our research. We are determined to put our resources where there is the broadest benefit. Over the next few years the laboratory's development work is likely to become more focussed. Our efforts are directed towards the detection of chromosome abnormalities in very small numbers of cells. Such analysis is important in such diverse situations as the assessment of residual disease in leukaemia, preimplantation chromosome analysis using one or two cells biopsied from an early embryo or detection of fetal chromosome abnormalities nondnvasively using 10-20 fetal erythroblasts isolated from a maternal blood sample.
Work
Keeping up-to-date with the latest is especially important For anyone in the medical field. New treatments, new drugs, new knowledge. Genetic knowledge and its link with disease is growing especially rapidly and our Genetic Counselling and Cancer Workshops have received an enthusi astic response.
In 1995 the VCGS initiated weekend workshops to provide medical and other healthcare professionals with the very latest information on scientific developments in the field of inherited cancers. Information covered the latest developments in cancer genetics including gene mapping and mutation detection, especially in breast and colon cancers. The workshops also highlighted the importance of a detailed family pedigree, communication techniques, and ethical and counselling issues affecting families by inherited cancers. Up to 30 people took part in each workshop, participating in interactive learning sessions and small group sessions. Videotaped examples of genetic counselling sessions were also viewed. These informative videos are available from the VCGS. Each participant also received comprehensive reading material to support the new information under discussion. A booklet of papers presented at the workshops is planned and workshops are continuing in 1997. The VCGS is grateful for the support of the Anti-Cancer Council of Victoria and their contribution towards funding of these workshops.
Louise Hills looking for metaphase chromosomes
Page 45
Page 46
Post GFaJuiLate Diipioma in Genetic C ©nnse mg
Another initiative of the VCGS has been the introduction of the Victorian Graduate Diploma in Genetic Counselling. Our first fulhtime students have now completed their course which includes lectures in core subjects, meetings and activities at the Murdoch Institute; and a four week practicum. This practicum reinforces the theoretical counselling and scientific skills learned during the course
Convened by Leslie Sheffield and Margaret Sahhar from the VCGS, the course is offered by the University of Melbourne (Faculty of Medicine) through the Murdoch Institute. Our present students are graduates in the field of science, social work and nursing. Skills and knowledge are shared by convenors of similar courses in other Australian states and we also offer supervised practical sessions for interstate students. At the start of 1997 we have eleven new students, and three continuing part'timers. We value the part-time students who bring practical skills and knowledge to the course from their employment.-especially in our interactive sessions. Course Outline. The diploma fulfils Part One of a two part process of qualification and certification as a genetic counsellor in Australia. Teaching staff are drawn from the scientific, medical genetics, social work and genetic counselling staff of the Institute. The teaching model is small group tutorial-based, using self-directed learning, and is based on the medical course developed at McMaster University, Hamilton, Ontario, Canada. The core subjects are genetics, counselling, and a four week supervised placement with a recognised genetics unit. Counselling subjects are offered in four modules (two per semester), and include training in counselling skills, interviewing in genetic counselling and genetic counselling in the community. Page 47
YOUR GENES AND YOUR INSURANCE The Murdoch Institute believes that everyone has a right to privacy with regard to their genetics, as with any other personal feature of family life. Insurance companies are beginning to propose that results of genetic tests should be made available to them. We promise our patients and their families that no genetic test information will be provided by us to insurance companies or employers. When people come for a test, it is to improve their chances of health. Foar of having test results divulged could reduce the likelihood of preventing serious diseases such as cancer. None of us control our genes ' we inherit them at random from our parents and we do not want to see the creation of a "genetic underclass". We take great precautions to make sure that families and their doctors are the only ones who can see genetic data from the Murdoch Institute, and we expect to keep it that way without the need for legislation.
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OUIF
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The Institute thanks all those listed below for their generous financial support during 1996 Mr dr Mrs G. Handbury Mrs J. Cal vert'Jones Mrs A. Kantor Dame Elisabeth Murdoch Mr dr Mrs M. Calvert'Jones Miss J. Kantor The Jack Brockhoff Foundation Mrs J. Roxburgh The Miller Foundation International Ladies Group AW Tyree Foundation Tattersalls ' The Estate of the late George Adams Uncle Bobs Club A very successful "Christmas Friends of the Murdoch Institute Tree Festival" was held in JB Were dr Sons Charitable Fund December by the Friends of Mr dr Mrs J.K. Little the Murdoch Institute. Mrs ANZ Banking Group Anne McFarling holds one of Dr J.M. Gooch the trees offered for sale. Mr G.R. Stephenson Mrs N. Gantner Johnson dr Johnson Mr G.E. Heeley Professor D. Penington William Angliss Charitable Fund Australian Recreation and Education Tours Mr S.F. dr Mrs M.G.Cooley MrW.H. Hodgson Professor Bob Williamson Mr B.E. Badman Mrs S.F. Kimpton Elmore Apex Club ' supporters of the Ms C. Craig Friedreich’s ataxia work of the gene Beta Sigma Phi (Ballarat) therapy team
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Scientific Hospital Supplies Australia Greyhound Promotions Pty Ltd Mr 1. Mrs T. Bryant Dr J. MacGill Mrs M. Stewart Mr W.C. &■ Mrs M.J. Wills Mrs B.M. Baillieu Mr E. Baillieu Mrs H.J. Daw Mr B.M. ^ Mrs K.M. Fraser Mrs R. Cunningham Ms S. O'Donnell J. Hawkes Mr &- Mrs L.R. Mills Dr T Nakura Mrs G.A. Grimwade D &- M Jewellery Mr A.C. gr Mrs V.A. Walsh
In memory of Emma Ritchie:
Mr R. Aldridge Mr K. Troon Mrs S. Hayden (K^mart) Mrs C. Maskiell Mrs B. Villinger Mr dr Mrs Louisotto Mrs J. Bowyer Mr dr Mrs Beckman Mr L. dr Mrs A.J. Cupit Mrs K. Fenwick Mrs E. Brown Mr J. dr Mrs T Sutton Mr A. dr Mrs H. Gunn Mr W. Troon Mrs R. Troon Mrs J. Fraser Monday Card Ladies Miss J. Ikin Mrs D. Haslem
In memory of Christopher Arthur Spence: Mr F.G. Dyett The Victorian Bar In memory of Shaun Nicholas Martin: Dr dr Mrs C.N. De Garis Mr dr Mrs Redman Mr D. dr Mrs J. Anderson Mr dr Mrs Martin Mr L. dr Mrs M. Buzzard Mr dr Mrs G.H. Fewster In memory of Brian Lange: Mrs R. Lazurus Staff of St. Margaret's School' Berwick Mr K. dr Mrs H. Nunn Mrs E. Luxton MrA. drMrsH. Dart Mr J.R. Payne Mr R. Rampling Mr C. dr Mrs G. Wilks Professor Williamson receives a cheque Ron Davies dr Son from Mrs Joan Roxburgh and members of the executive of the Uncle Bobs Club.
5:
mMoc li Insttittiife
111'
St an
Stephen Wilcox, B.Sc. (Hons.), Ph.D. Kathy Williamson, B.Sc. (Hons.), Ph.D. Tracy EvanS'Whipp, B.Sc. (Hons.) Ph.D. Daniel Strausak, Lie. phil. nat. (Switz.) Ph.D.
Clinical Fellows
Scientific Director Robert Williamson, Ph.D.,FRCPath., Hon. MRCP, Hon.M.D. (Turku)
Business Manager Anne Cronin, B.Sc., B.Bus.(Acc.), A.S.A., C.P.A.
Laboratory Manager Barry Holt, B.App.Sci.(M.T), A.A.I.M.L.S.
Scientists (Senior) Jim Camakaris, B.Sc.(Hons.), Ph.D. K.H. Andy Choo, B.Sc.(Hons.), Ph.D. HanS'Henrik Dahl, Ph.D. Julian Mercer, B.Sc.(Hons.), Ph.D. Donald Newgreen, B.Sc.(Hons.), Ph.D.
Senior Research Fellows
Ilf
Susan Forrest, B.Sc.(Hons.), D.Phil.(Oxon.), B.Bus.(Admin.). David Thorburn, B.Sc.(Hons.), Ph.D. Jane Halliday, B.Sc.(Hons.), Ph.D.
Clinical Scientists Agnes Bankier, M.B., B.S., F.R.A.C.P. Maureen Cleary, M.B. ChB. M.R.C.P. John Rogers, M.B., B.S., D.C.H., F.R.A.C.P. Mac Gardner, M.B., Ch.B., M.Sc. F.C.C.M.G. Les Sheffield, B.Med.Sci., M.B., B.S., M.Sc., D.C.H., F.R.A.C.P.
Postdoctoral Fellows
i:
Susan G. Bevan, Ph.D. Michael Cancilla. B.Sc.(Hons.), Ph.D. Sharon La Fontaine. B.Sc. (Hons.), Ph.D. Richard Saffery, B.Sc. (Hons.), Ph.D. Helen Trowell, B.Sc.(Hons.), Ph.D. Page 51
Martin Delatycki, M.B., B.S., FR.A.C.P. (Part 1) Catherine Rose, M.B., B.S. Karen Dunn, B.Med.Sci. M.B., B.S. Heidi Peters, M.B., B.S.
Michael Petris B.Sc.(Hons.) Michael Theophilos B.Sc.(Hons.) Louise Wangerek, B.Ag.Sci. Sarah White, B.Sc. (Hons.) Kumaran Narayanan, B.Sc. (Hons) Swari Setianingsih, M.D.
Technical Assistants
Moria Graham Roseanna Bhagwandas Alison Blake Scientific Officers and Research Assistants Evelyn Boyer Robyn Breslin Hilary Brooks, B.Sc. (Hons.) Sophie Gazeas Marjorie Crawford, A.R.M.I.T Michelle Guneratne Elizabeth Earle, A.A.I.M.L.S. Tiffany Symes Stephen Firth, M.App. Sci. Rosario Reyes Kerry Fowler, M.App.Sci., M.Sc. Matthew Newman Andrew Grimes, B.App.Sci. Blanche Dekker Joanne Hill, B.App.Sci. Tal Rapke Sharon Horton, B.Sc., Grad.Dip.Diet. Wendy Hutchison, B.App.Sci.(App.Biol.) Danielle Irvine, B.Sc. (Hons.) Denise Kirby, B.Sc.(Hons.) Henny Miller Joseph Minichiello, M.Sc. Erin Oldaker B.App.Sci. Amelia Osborn, B.App.Sci. Damien Paris, B.Sc. (Hons.) Jenny Paynter, B.Sc.(Hons.) Janet Shaw, B.Sc. (Hons.) Kellie Tainton, B.Sc. (Hons.) Carole Webley Stewart Beattie, B.Sc. (Hons)
Ph.D. Scholars Loreta Ambrosini, B.App. Sci. Alyssa Barry, B.Sc. (Hons.) Desiree Du Sart, B.App.Sci. Catherine Hearn, B.Sc. (Hons.) Emily Howman, B.Med. Lab. Sci. Damien Hudson, B.Sc.(Hons.) Paul Kalitsis, B.Sc. Richard Kerr, B.Sc. (Hons.) Paul Lockhart, B.Sc. (Hons.) Laraine Peters, B.Sc.(Hons.)
Dietitian Dorothy Francis, SRD Administration Accountant: Hilary Lloyd, B.Ec., A.S.A. (to 9-5^96) Viren Abeysinghe, M.B.A., F.C.M.A. Personnel Assistant: Debbie Zombolas Personal Assistant to the Director: Margot Latham, B.Sc. (to 20'9'96) Julie Foletta, B. Ed.
Receptionists: Fiona Keltic Olga Wilson Vicki Hurt
Secretaries: Debbie Davis Kristine Yeomans
Photography/Graphic Design Michele Winsor Kati Hidegh Page 52
Victorian Clinica Genetics S ervices
Staff
Executive Director: Robert Williamson, Ph.D.,FRCPath., Hon. MRCP, Hon.M.D. (Turku)
ClinicaJ Geneticists Agnes Bankier, M.B., B.S., F.R.A.C.P. John Rogers, M.B., B.S., D.C.H., F.R.A.C.P. Les Sheffield, B.Med.Sci., M.B., B.S., M.Sc., D.C.H., FR.A.C.P Mac Gardner, M.B., Ch.B., M.Sc., F.C.C.M.G.
Anne Robertson, B.Sc., H.G.S.A.C.C. Marie Thorpe, B.Sc.(Hons.) Tacy Vankuyk, B.Sc. (Hons.) Cathryn Vaux, B.Sc., Grad. Dip. Genetic Counselling Lucille Voullaire, M.Sc., H.S.S.A.C.C. Sarah Nouri, B.Sc. (Hons.) Wall Drummond, Dip. Basic Med. Sci. Lorna Webber, TP.T.C., B.Sc. (Hons.), Ph.D.
Jo Wells Mary'Anne Young, S.R.N. Susan Clement, M.Sc., B.A.
Technical Assistants Lynda Phillips Ian Brooks Thomas Milovac, Ass. Dip. App. Sci. (Lab. Tech.) Fernando Garcia
Anne Cronin, B.Sc., B.Bus.(Acc.), A.S.A., C.P.A.
Metabolic Physician il:
DNA Diagnosis ^ Scientists Susan Forrest, B.Sc.(Hons.), D.Phil.(0 xon .), B.Bus.(Admin.) ^ Scientist in charge Ivan Biros, B.Sc., Ph.D. Janice Brasch, B.Sc.(Hons.), M.Sc. Karina Forshaw, B.App.Sci. Kathy Garofalo (Trainee) Steven Nasioulas, B.Sc. (Hons.) Andrea Twomey, B.Sc.(Hons.) Jenny Douglas, B.Sc. (Hons.) Jean Hendy, B.Sc., FA.l.M.L.S.
Laboratory Assistant Bozena Jezierski
Neonatal Screening Laboratory ' Scientists
Cytogenetics - Scientists
Louise Hills, B.Sc.
Ralph Oertel, B.Sc., H.G.S.A.A.C. Vida Petrovic, B.Sc., H.G.S.A.A.C. Page 53
Susan Mansie, S.R.N., B.S.W. Margaret Sahhar, B.A., Dip.Soc. Studies
Business Manager
Maureen Cleary, M.B., Ch.B., M.R.C.P. (UK)
Howard Slater, B.Sc., Ph.D., Dip.R.C.Path., H.G.S.A.C.C. ^ Scientist in charge Selena Bourke, B.Sc. Trent Burgess, B.Sc.(Hons.) Melissa Curtis, B.Sc. Sue Dale, B.Sc.(Hons.) Julie Davies, B.Sc., H.G.S.A.A.C. David Francis, B.Sc.(Hons.) Julie Garratt, B.Sc.(Hons.)
Social Workers
1
Laboratory Manager Barry Holt, B.App.Sci.(M.T), A.A.I.M.L.S.
Project Officer ^ Maternal Serum Screening Kelley Gardner, R.N. Assoc. Sci. (Hons.) Robyn Forbes
Ivan Francis, B.Sc., Dip.Comp.Sci. ' Scientist'inCharge Leonard Bonacquisto, B.Sc.(Hons.), Dip.Comp.Sci. Mona ELMasri Nella Napolitano Nick Tzanakos, B.App.Chem. Kate Milkins, B.Sc.
Administrative Assistant
Genetic Clinic Co-ordinators / Genetic Counsellors
Cathy McMillan (to 6'3'96) Christine Keenan
Sue Casanelia, S.R.N. Clara Gaff, B.Sc.(Hons.), Ph.D. Margaret Olsen, Dip .App. Biol., Dip. Ed. Ann Robertson, S.R.N. Linda Warwick, S.R.N.
Sharon Vandersluis
Secretary Lorraine White
Accounts Clerk
Computer Support Shilpa Shah
Page 54
'i 'i
Si ='1
of PuilLlicafions
In press previous report now published
Rudzki, Z, SE Rogers, LJ Sheffield, and JV Lloyd. Detection of carriers of haemophilia A: use of bioas' says and restriction fragment length polymorphisms (RFLP). Aust. N.Z. J of Med. 26: 195'205, 1996.
Berkovic, SF, A McIntosh, RA Howell, A Mitchell, LJ Sheffield, and JL Hopper. Familial temporal lobe epilepsy: a common disorder identified in twins Ann Neurol
Youil, R, B Kemper, and RGH Cotton. Detection of 81 of 81 known mouse beta'globin promoter muta^ tions with T4 endonuclease VII ' the EMC method. Genomics 32: 431 '435, 1996.
40: 227^235, 1996. Brown, NL. SR Barrett, J Camakaris, BTO Lee, and DA Rouch. Molecular genetics and transport analysis of the coppet'resistance determinant (pco) from Escherichia coli plasmid PRJ1004. Mol Microbiol 17: 1 153'1 166, 1995.
Published and accepted for publication since 1995 report Allotey, R, R Twells, C Cemal, B Schleich, J Weissenbach, M Pook, R Williamson, and S Chamberlain. The spinocerebellar ataxia 2 locus is located within a 3'cM interval on chromosome 12q23'24.1. Am J Hum Genet 57: 185'188, 1995.
Distante, S, S Nasioulas, Somers. GR, DJS Cameron, MA Young, SM Forrest, and RJM Gardner. Familial adenomatous polyposis in a 5 year old child: a clinical, patho' logical and molecular genetic study. J Med Genet 33: 157' 160, 1996.
Apostolova, MD, KHA Choo, AE Michalska, and C Toyama. Analysis of possible protective role of metah lothionein in streptozotocin ' induced diabetes using metallothionein'null mice. Trace Elements in Med &Bio. (in press).
Duband, J'L, F Monier, M Delannet, and D Newgreen. Epithelium^mesenchyme transitions during the development of the neural crest. Acta Anat 154: 63'78, 1995. Gupta, SD, BTO Lee, J Camakaris, and HC Wu. Identification of cutC and cutF (nIpE) genes involved in copper transport in Escherichia coli. J Bacteriol 177- 4207' 4215, 1995.
Bankier, A. “Genetic counselling." In Clinical Paediatric Surgery., Blackwells, (in press). Bankier, A, "Syndrome identification.” In Atlas of Pediatric Oral Medicine and Oral Pathology., Chapman' Hall, (in press).
Jones, LN, DJ Peet, DM Danks, AP Negri, and DE Rivett. Hairs from patients with maple syrup urine disease show a structural defect in fiber cuticle. J Invest Dermatol
Berg, K, B Brambati, N Fujiki, EK Ginter, BM Knoppers, B Modell, 1C Verma, and R Williamson. The control of hereditary diseases ' report of a WHO scientific group. World Health Organisation, Geneva,
106: 461'464, 1996.
1996.
Newgreen, DF, and J Minichiello. Control of epitheliomesenchymal transformation. II. Cross'modulation of cell adhesion and cytoskeletal systems in embryonic neural
Bevan, SG, M Southey, JE Armes, DJ Venter, and DF Newgreen. Spatio'temporally exact cDNA libraries from quail embryos: a resource for studying neural crest development and neurocristopathies. Genomics
cells. Dev Biol 176: 300'312, 1996.
38: 206'214, 1996.
Rahman, S, RB Blok, H'HM Dahl, DM Danks, DM. Kirby, CW Chow, J Christodoulou, and DR Thorburn. Leigh syndrome: clinical features and biochemical and DNA abnormalities. Annals of Neurol 39: 343'351, 1996.
l! !i'
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Ramus, SJ, and RGH Cotton. Polymorphism in the 3' untranslated region of the phenylalanine hydroxlase gene detected by enzyme mismatch cleavage: evolution of haplotypes. Hum Genet 96: 741'743, 1995.
Boulton, M and R Williamson. General practice and new genetics: what do general practitioners know about community carrier screening for cystic fibrosis? Public Understand Sci. 4: 255'267, 1995. I
Boulton, M, C Cummings, and R Williamson. The views of general practitioners on community carrier screening for cystic fibrosis. Brit. J. Gen. Practice 46: 299'301, 1996. Page 56
Boulton, M, C Cummings, E Mayall, and R Williamson. The value of a video as a means of informing and reassuring autonomous CF carriers identified by a communis ty screening programme. Health Education J 55: 203-214, 1996.
Cotton, RGH, MA Knight, M Southey, C Anderson, A Tesoriero, S Brown, K Jennings, D Venter, J Hopper, and RYsuil. A comparative study between enzyme mismatch cleavage (EMC) and direct sequencing to detect for mutations within the BRCAl gene. Am Hum Genet 59: abs 1466, 1996.
Caplan, NJ, EW Alton, PG Middleton, JR Dorin, BJ Stevenson, X Gao SR Durham, PK Jeffery, ME Hodson, C Coutelle, L Huang, DJ Porteous, R Williamson, and DM Geddes. Liposome-mediated CFTR gene transfer to the nasal epithelium of patients with cystic fibrosis. Nature Med 1: 39-46, 1995.
Cotton, RGH. Locus specific mutation detection. In: laboratory protocols, ed. U. Landegren. Oxford: OUP, 5-7, 1996.
Caplen, NJ, E Kinrade, F Sorgi, X Gao, D Grunert, D Geddes, C Coutelle, L Luang, EWFW Alton, and R Williamson. In vitro liposome-mediated DNA trans fection of epithelial cell lines using the cationic liposome DC-Chol/DOPE. Gene Therapy 2: 603-613, 1995. Carvajal, JJ, MA Pook, K Doudney, R Hillerman, D Wilkes, S Al-Mahdawi, R Williamson, and S Chamberlain. Friedreich's ataxia: a defect in signal transduction? Hum. Mol. Genet. 9: 1411-1419, 1995. Carvajal, JJ, MA Pook, M dos Santos, K Doudney, R Hillermann, S Minogue R Williamson, JJ Hsuan, and S Chamberlain. The Friedreich's ataxia gene encodes a novel phosphatidylinositol-4-phsphate 5-kinase. Nature Genet. 14: 157-162, 1996. Clemens, M, JT Martsolf, JG Rogers, Mowery-Rushton, U Surd, and E McPherson. Pitt-Rogers-Danks syndrome: The result of a 4p microdeletion. J Med Genet 66: 95-100, 1996. Cole, WG, D Chan, CW Chow, JG Rogers, and JF Bateman. Disrupted growth plates and progressive deformities in osteogenesis imperfecta due to the substitution of glycine 586 by valine in the alpha2(I) chain of type 1 collagen. J Med Genet 33: 968-971, 1996. Cotton, RGH and U Landegren. Interest rising in mutation detection. Genome Digest 2: 1-4, 1995. Cotton, RGH, C Scriver, and VA McKusick. HUGO's second meeting on locus specific mutation databases. Genome Digest 3: 14-15, 1996. Cotton, RGH, C Scriver, and VA McKusick. Locus specific databases a resource. Genome Digest 3: 6-10, 1996. Page 57
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Cotton, RGH. Detection of unknown mutations in DNA - a catch 22. Am J Hum Genet 59: 289-291, 1996. Cotton, RGH. Mutation databaise initiative underway. Human Genome News 8: 13-14, 1996. Coutelle, C, and R Williamson. Liposomes and viruses for gene therapy of cystic fibrosis. J. Aerosol Med. 9: 79-88, 1996. Dahl, H-HM, J Fitzgerald, and R lannello. “The sperm-specific pyruvate dehydrogenase Ela genes.” In Alpha-keto acid dehydrogenase complexes., ed. TE. Roche and R.A. Harris M.S. patel. Switzerland: Birkauser Verlag Basel, 213-226, 1996. Davies, GE, CM Howard, MJ Farrar, MM Coleman, LB Bennett, LM Cullen, RKH Wyse, J Burn, R Williamson, and AM Kessling. Congential heart defects in trisomy 21: aissociation with genetic variation in the COL6A1 region. Ann. Hum. Genet. 59: 253-269, 1995. Delatycki, M, and LJ Sheffield. Familial heterotaxia: What is the inheritance in this family? Am J Med Genet, (in press). Delatycki, M, MA Cleary, A Bankier, PN McDougall, JS Ahluwalia, CW Chow, and CM CookeYarborough. A maternally transmitted neonatal progeroid syndrome with prominent genitourinary and gas trointestinal features: X linked or mitochondrial aetiology? J Med Genet, (in press). Delatycki, M, S Nasioulas, K Forshaw, and S Forrest. 2 novel mutations in exons 5 and 15 of the adeno matous polyposis coli (ape) gene. Hum Mut, (in press). du Sart, D and KHA Choo. Simultaneous fluorescence in situ hybridisation and anti-centromere antibody staining of normal and "stretched" metaphase chromosomes. In Molecular Biotechniques, ed. R. Rapley and J.M. Walker. Tbtowa, NJ, USA: Humana Press, (in press). Fitzgerald, J, H-HM Dahl, IB Jakobsen, and S Easteal. Evolution of mammalian x-linked and autosomal Pgk and Pdh Ela-subunit genes. Mol Biol Evol 13: 1023-1031, 1996. Page 58
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Franks, S, D White, C Gilling^Smith, A Carey, D Waterworth, and R Williamson. "Hypersecretion of androgens by polycystic ovaries: the role of genetic factors in the regulation of cytochrome P450cl7a.” In Bailliere's clinical endocrinology and metab^ olism ^ polycystic ovary syndrome., ed. HS Jacobs Guest Editor, 1996.
Hillerman, R, CG See, M Pook, D Wilkes, J Carvajal, K Doudney, R Williamson, and S Chamberlain. Physical evidence for the position of the Friedreich's ataxia locus FRDA proximal to D9S5. Cytogenet Cell Genet 71: 214-216, 1995.
Freckmann, M^L, DR Thorburn, DM Kirby, KR Kamath, J Hammond, X Dennett, and J. Christodoulou. Mitochondrial electron transport chain defect presenting as hypoglycemia. J Pediatr, (in press).
Johansen, PA, M Zhu, K San Karan, 1 Jennings, RGH Cotton, and DM Kuhn. Tryptophan hydroxylase protein kinase interactions - dissociation of phosphorylation and activation by protein kinase A J Neurochem 65: 882-885, 1995.
Gardner, RJM and GR Sutherland. Chromosome abnormalities and genetic counsel ing (2nd edition). New York, Oxford University Press, 1996. Gharani, N, DM Waterworth, S Batty, D White, D Gilling-Smith, GS Conway, M McCarthy, S Franks, and R Williamson. Association of the steroid synthesis gene CYPl la with polycystic ovary syndrome and hyperandrogenism. Hum Mol Genet, (in press). Giunta, C, R Youil, D Venter, CW Chow, G Somers, A Lafferty, B Kemper, and RGH Cotton. Rapid diagnosis of germline p53 mutation using the enzyme mismatch cleavage method. Diagn Mol Path, (in press). Halhday, JL and C Webley. Report on prenatal diagnostic testing in Victoria Murdoch Institute. ISSN 1327-7618, 1996. Hargest, R and R Williamson. Expression of the APC gene after transfection into a colonic cancer cell line. Gut 37: 826-829, 1995. Hargest, R and R Williamson. Prophylactic gene therapy for cancer. Gene Therapy 3:97-102,1996. Hart, S, E Mayall, M Stern, E Munkonge, A Frost, L Huang, M Vasilliou R Williamson, EWFW Alton, and C Coutelle. The introduction of two silent muta tions into a CFTR cDNA construct allows improved detection of transgeni c message in gene transfer experiments. Hum Mol Genet 9: 1597-1602, 1995. Hart, SP RP Harbottle, R Cooper, A Miller, R Williamson, and C Coutelle. Gene delivery and expression mediated by an integrin-binding peptide. Gene Therapy 2: 552-554, 1995.
Kedinger, M, and D Newgreen. The gut and enteric nervous system. In: Birth Defects: Perspectives from Contemporary Development Biology, ed. P Thorogood (ed). UK: John Wiley 6- Sons, (in press). Lam, WK, MA Clary, JH Walter, JE Wraith, and IB Sardharwalla. Histidinaema: a benign metabolic disorder. Arch Dis Child 74: 343-346, 1996. Liu, J, Y Liu, AE Michalska, KHA Choo, and CD Klaassen. Distribution and retention of cadmium in metallothionein 1 and 11 null mice. Toxicol Appl Pharmacol 136: 260-268, 1996. Liu, J, Y Liu, AE Michalska, KHA Choo, and CD Klaassen. Metallothionein plays less of a protective role in cadmium-metallothionein-induced nephrotoxicity than in cadmium chloride-induced hepatotoxicity. J Pharmacol Expt Therapeutics 276: 1216-1223, 1996. Loesch, DZ, V Petrovic, D Francis, R Oertel, and H Slater. Reduction of CGG trinucleotide expansion from mother to offspring in seven fragile X families. Clin Genet (in press). Mansie, S, LJS Sheffield, and S Forrest. Selection for presymptomatic testing for Huntington's disease: who decides? A reply from the Victorian Clinical Genetics Service, Murdoch Institute, Melbourne, Australia. J Med Genet 33: 1051-1054, 1996. Matthews, D, L Fry, A Powles, J Weber, M McCarthy, E Fisher, KE Davies, and R Williamson. Evidence that a locus for familial psoriasis maps to chromosome 4q. Nature Genet 14: 231-233, 1996. Matthews, D, L Fry, A Powles, J Weissenbach, and R Williamson. Confirmation of genetic heterogeneity in familial psoriasis. J Med Genet 32: 546-548, 1995. Mercer, JFB and J Camakaris. Inherited disorders of copper transport in mammalian systems. In: Metal Ions in Gene Regulation, ed. Chapman and Hall (in press). Mercer, JFB and DM Danks. Disorders of copper transport. In: Metabolic and molecular basis of inherited disease., ed. A1 Baudet CR Scriver, WM Sly and D. Valle. New York: McGraw-Hill (in press).
Page 59 Page 60
ll Mercer, JFB. Gene regulation by copper and the basis for copper homeostasis. Nutrition (in press)
Page, SL, J Shin, J Han, KHA Choo, and LG Shaffer. Breakpoint diversity illustrates distinct mechanisms for Robertsonian translocation formation. Hum Mol Genet 5: 1279-'1288, 1996.
Mercer, JFB. Genetic disorders of copper metabolism and the dual nature of copper in biology. In; Proceedings of international wo rkshop on copper. Copper: Essentiality and Toxicity (in press).
Petris, M, JFB Mercer, JG Culvenor, P Lockhart, PA Gleeson, and J Camakaris. Ligand-regulated trans port of the Menkes P-type ATTPase efflux pump from the transGolgi to the plasma membrane; a novel mechanism of regulated trafflking. EMBO J 15: 6084-6095, 1996.
Mercer, JFB. Menkes disease and animal models. Am J Clin Nutrition (in press). Moores, C, JG Rogers, IM McKenzie, and TCK Brown. Anaesthesia for children with mucopolysaccharidoses. Anaesth Intens Care 24: (in press). Nasioulas, S, L Sheffield, S Mansie, and S Forrest. Modified protocol for the detec tion of the GAG repeat expansion in Huntington's disease and application to a predictive testing protocol. Mol Diag (in press). Newgreen, DF and J Minichiello. Control of epithdio-mesenchymal transformation 1. Events in the onset of neural crest cell migration are separable and inducible by protein kinase inhibitors. Dev Biol 170: 91-101, 1995. Newyeen, DF and L Hartley. Extracellular matrix and adhesive molecules in the early development of the gut and its innervation in normal and spotting lethal rat embryos. Acta Anat 154: 243-260, 1995. Newgreen, DF and RSE Kerr. Adhesion inhibiting molecules in neural crest morpho genesis. In; Interplay of genetics and physical processes in the development of biologPte Ud"''l5^2f 19°9T“ Publishing Co.
Newgreen, DF, B Southwell, L Hartley, and IJ Allan. Migration of enteric neural crest cells in relation to the growth of the embryonic avian gut. Acta Anat, (in press). Newport, M, C Huxley, S Huston, C Hawrylowicz, BA Oostra, R Williamson, and M Levin. A mutation in the interferon-gamma receptor gene causes susceptibility to mycobacterial infection in man. New Eng J Med. 335: 1941-1949, 1996. Newport, M, M Levin, J Blackwell, MA Shaw, R Williamson, and C Huxley Evidence for exclusion of a mutation in NRAMP as the cause of familial disseminatmycobacterial infection in a Maltese kindred. J Med Genet 32; 904-906, Page 61
Pitt, JJ, R Hawkins, L Warwick, M Cleary, and DR Thorburn. Low excretion of metabolites in succinic semi-aldehyde deficiency. J Inher Metab Dis, (in press). Ramus, SJ and RGH Cotton. Single tube chemical cleavage of mismatch: successive treatment with hydroxlamine and osmium tetroxide. BioTechniques 21: 216-220, 1996. Riley, B, M Mogudi-Carter, T Jenkins, and R Williamson. No evidence for linkage of chromosome 22 markers to schizophrenia in Southern African Bantu-speaking families. Am J Med Genet 67: 515-522, 1996. Riley, BP and R Williamson. Non-parametric analysis of chromosome 6p24-22 marker data and schizo phrenia in Southern African Bantu-speaking families. Psychiatric Genet 67; 580-594, 1996. Riley, BP, E Tahir, S Rajagopalan, M Mogudi-Carter, S Faure, J Weissenbach, T Jenkins, and R Williamson. A linkage study of the N-methyl-D-aspartate receptor subunit gene loci and schizophrenia in Southern African Bantu-speaking families. Psych Genet, (in press). Riley, BP, S Rajagopalan, M Mogudi-Carter, T Jenkins, and R Williamson. No evidence for linkage of chromosome 6p markers to schizophrenia in Southern African Bantu-speaking families. Psychiatric Genet 6; 41-50, 1996. Riley, M and JL Halliday. Congenital malformations in Victoria, 2983-1994. Report for Victorian Department of Human Services June; 1996. Sato, M, MD Apostolova, N Kodama, J Yamaki, M Hamaya, AE Michalska, KHA Choo, and C Tohyama. Susceptibility of metallothionein-deficient mice to paraquat. Environ Toxicol Pharmacol (in press). Slater, H, C Vaux, M Pertile, and V. Petrovic. Prenatal diagnosis of Prader-Willi syndrome - uniparental disomy and the significance of residual trisomy 15. Pren Diag (in press). Smooker, PM and RGH Cotton. The molecular basis of dihydropterine reductase deficiency. Hum Mut 5; 279-284, 1995.
Page 62
i Somers, GR, H Slater, D MacDonald, H Ekert, MC Southey, CW Chow, and DJ Venter. Tcell lymphoblastic lymphoma and an atypical myeloproliferative disorder associated with t(8; 13)(p21;ql4). Paed Path Lab Med (in press).
mammalian niitochondrial and universal translation systems: a novel approach towards correction of a genetic defect. Gene 169: 251'255, 1995. Williamson, R. Medical ethics, teaching and the new genetics. J Med Ethics 22: 325'326, 1996.
Takayama, K, DM Danks, EP Salazar, JE Cleaver, and CA Weber. DNA repair char^ acteristics and mutations in the ERCC2 DNA repair and transcription gene in a tri^ chothiodystrophy patient. Hum Mut, (in press). Theophilos, MB, DW Cox, and JFB Mercer. A mutation in the 8th transmembrane domain of the Wilson disease gene homologue (Atp7b) in the toxic milk mouse: a murine model of Wilson disease. Hum Mol Genet 5: 1619^1624, 1996. Thomas, MR, B Tutschek, A Frost, CH Rodeck, N Yazdani, 1 Craft, and R Williamson. The time of appearance and disappearance of fetal DNA form the maternal circulation. Prenat Diagn 15: 641 •'646, 1995. Toder, R, RW O'Neill, J Wieberg, L Voullaire, and JAM Graves. Comp arative chromosome painting between two marsupials: origins of an XX/XY Y2 sex chro' mosome system. Mammalian Genome (in press).
Williamson, R. Towards non'invasive antenatal diagnosis. Nature Genet 14: 239'240, 1996. Yano, S., L. Sweetman, D.R. Thorburn, S. Modifi, and J.C. Willilams. A new case of malonyl coenzyme A decarboxylase deficiency presenting with cardiomyopathy. Eur J Pediatr (in press). Young, H., D. Ciampoli, B. Southwell, and D.F. Newgreen. Origin of the interstitial cells of Cajal in the mouse intestine. Dev Biol (in press) Zheng, H, J Liu, KHA Choo, AE Michalska, and CD Klaassen. Methallothionein'l and '11 knock'out mice are sensitive to cadmium'induced liver mRNA expression of cjun and p53. Toxicol Appl Pharmacol 136: 229'235, 1996.
Tohyama, C, M Satoh, N Kodama, H Nishimura, KHA Choo, AE Michalska, Y Kanayama, and A Naganuma. Reduced retention of cadmium in the liver of metah lothioneinmull mice. Environ Toxico Pharmacol 1: 213'216, 1996. Treacy, E, JJ Pitt, K Sellar, G Thompson, S Ramus, and RGH Cotton. In vivo dis' posal of phenylalanine in mild phenylketonuria: a case study of two siblings J Inher Metab Dis 19: 575'602, 1996. Treacy, EP, R YduH, S Forrest, M Knight, RGH Cotton, and J Cashman. A^G substi^ tution in the FM03 gene, molecular basis for altered pharmacokinetics, association with trimethylaminuria(?). Am Hum Genet 59: abs 1 189, 1996. Twells, R, X Weiming, D Ball, RAllotey, R Williamson, and S Chamberlain. Exclusion of the neuronal nitric oxide synthase gene and the human achaete-scute homologue 1 gene as candidate loci for spinal cerebellar ataxia 2. Am. J. Hum. Genet. 56: 336'337, 1995. Wheeler, VC, C Prodromou, LH Pearl, R Williamson, and C Coutelle. Synthesis of a modified gene coding for human ornithine transcarbamylase for expression in Page 63
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^TT TT
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MURDOCH INSTF
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FOR RESEARCH INTO BIRTH DEFECTS LIMITED
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A.C.N. 006 566 972 and its controlled entity Financial Statements and Reports 31st December 1996
Page 65
Page 66
I .Directors Report
The Directors present their report together with the SLCcounts of the Murdoch Institute for Research into Birth Defects Limited "the Company" and the consolidated accounts of the economic entity, being the Company and its controlled entity, for the year ended 31 December, 1996 and the auditors' report thereon
Directors The Directors of the Company in office at the date of this report are: Mr. L.G. Cox, B.Com., A.S.A., F.S.I.A. Chairman (Non-Executive Director)
Dr. G.L. Barnes, M.D., Ch.B., F.R.A.C.P. Non-Executive Director Age 55 Scientific Director, Royal Children's Hospital Research Foundation. Senior Gastroenterologist, Royal Children's Hospital. Dr. Barnes represents the Hospital on the Institute's Board. Director since 1986. Mrs. J. Calvert-Jones Non-Executive Director Age 58 Chairman of the Herald and Weekly Times Limited. Chairman of the Advisory Council for Children with Impaired Hearing (Vic). Mrs Calvert-Jones represents Cruden Investments Pty Ltd on the Institute's Board. Director since 1986.
I,:;
Age 58 Mr Cox is an investment banker and a director of various public and private nies and community groups. Director since 1986 - appointed Chairman 1993. Mr. W.H. Hodgson Deputy Chairman (Non-Executive Director) Age 67 Formerly Deputy Managing Director of the National Australia Bank Limited, an appointment which completed a long career with the Bank. Mr Hodgson is a Director of various public and private companies. Director since 1986. Professor J.A. Angus, B.Sc.(Hons), Ph.D. Non-Executive Director
Age 48 Professor Angus holds the Chair of Pharmacology at the University of Melbourne. He represents the National Health 6- Medical Research Council (NHMRC) on the Institute's Board. Director since 1991. Page 67
Mrs. L. Cattermole, B.Sc. Non-Executive Director Age 48 Chairperson of the Women's and Children's Healthcare Network. Director since 1995. Mr. 1. Davies, B.H.A., F.A.I.M., F.C.H.S.E. Non-Executive Director
Age 56 Chief Executive Officer, Women's and Children's Healthcare Network. Mr. Davies was formerly the Chief Executive Officer of the Royal Children s Hospital. Director since 1991. li!
Mr. J.A. Fitzgerald Non-Executive Director Age 65 Mr. Fitzgerald is a public relations consultant. He is Corporate Affairs adviser to some of Australia's largest corporations. Director since 1986. Page 68
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i Mr. P. Griffin, B.Comm.(Melb) Non'Executive Director Age 57 Chairman of the Institute's Finance Committee. Mr. Griffin is an investment banker and a director of N.M.Rothschild &- Sons (Aust) Pty Ltd Group companies, various public and private companies, industry and community groups. Director since 1993.
Professor R. Williamson, PhD., F.R.C. Path., Hon. M.R.C.P., Hon.M.D. (Turku) Scientific Director Age 58 Executive Director of the Victorian Clinical Genetics Services. Professor of Medical Genetics, University of Melbourne. Director since 1995.
Directors' Meetings Mr. J.S. Guest, A.M., O.B.E., V.R.D., B.Sc., M.B., B.S., F.R.C.S., F.R.A.C.S. Non-Executive Director Age 80 Mr. Guest is a distinguished Melbourne surgeon and the Chairman of the Jack Brockhoff Foundation. Director since 1986. Mrs. I. McFarling Non-Executive Director Age 59 Mrs. McFarling is a successful public relations advisor. She is the President of the Friends of the Murdoch Institute, the Fundraising Auxiliary of the Institute. Director since 1988. Mrs. J. Paterson Non-Executive Director Age 39 Appointed during 1996. Professor D.G. Penington, A.C., M.A., D.M., B.Ch., F.R.C.P., F.R.A.C.P., F.R.C.P.A. Non-Executive Director Age 67 Professor Penington was formerly the Vice-Chancellor, the University of Melbourne. He is a director of various public and private companies. Appointed during 1996.
Page 69
Professor G.W. Tregear, B.Sc., PhD., F.R.A.C.I. Non-Executive Director Age 56 Associate Director, Howard Florey Institute of Experimental Physiology and Medicine. Professor Tregear represents the NH&-MRC on the Institute's Board. Director since 1994.
The number of directors' meetings and number of meetings attended by each of the directors of the Company during the financial year are: Director Mr. L.G. Cox Mr. W.H. Hodgson Professor J.A. Angus Dr. G.L. Barnes Mrs. J. Calvert-Jones Mrs. L. Cattermole Mr. I. Davies Mr. J.A. Fitzgerald Mr. P. Griffin Mr. J.S. Guest Mrs. 1. McFarling Mrs. J. Paterson Professor D.G. Penington Professor P.D. Phelan Professor G.W. Tregear Professor R. Williamson
No of Meetings Attended 5 4 4 5
3 4 4 5 4 4 4 5 5 5 4 6
No of Meetings Held 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6
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ii,
Principal Activities The principal activities of the economic entity during the course of the financial year were to promote and undertake medical research into the understanding, prevention and treatment of birth defects, and to provide services for the diagnosis and treatment of genetic diseases and other birth defects. No significant change in the nature of these activities occurred during the year.
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Dividends The Company is a company limited by guarantee. As such it has no share capital, and no dividends are paid.
shown in the consolidated accounts) because of a contract made by the Company, its controlled entity, or a related body corporate with the Director or with a firm of which the Director is a member, or with an entity in which the Director has a sub' stantial interest.
Consolidated Result The consolidated operating profit of the economic entity for the financial year ending 31 December 1996 was $6,004,533. (1995 loss $546,520). No provision is required for taxation as the Company and its controlled entity are exempt from Income Tax.
Review of operations A review of the operations of the Company and its controlled entity during 1996 has been included in the Report of the Board. Dated at Melbourne this 23rd day of April 1997.
Significant changes in state of affairs
Signed in accordance with a resolution of the Directors:
In the opinion of the Directors there were no significant changes in the state of the economic entity's affairs during the financial year under review.
Events Subsequent to Balance Date There has not arisen in the interval between the end of the financial year and the date of this report any item, transaction or event of a material and unusual nature likely, in the opinion of the Directors of the Company, to affect significantly the oper' ations of the economic entity, the results of those operations, or the state of affairs of the economic entity, in subsequent financial years.
LAURENCE G. COX : Directors
ROBERT WILLIAMSON
Directors' Interests and Benefits Since the end of the previous financial year, no Director of the Company has received or become entitled to receive any benefit (other than a benefit included in the aggregate amount of remuneration received or due and receivable by Directors Page 71
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THE MURDOCH INSTITUTE FOR RESEARCH INTO BIRTH DEFECTS LIMITED AND ITS CONTROLLED ENTITY 'i \
Profit and Loss Accounts for the year ended 31 December 1996
Balance Sheets as at 31 December 1996 Note
Note
Consolidated
The Company
1996
1995
1996
1995
$
$
$
$
6,004,533
(546,520)
5,914,594
(556,439)
Operating Profit/(Loss) Income Tax
6,004,533
(546,520)
5,914,594
(556,439)
Accumulated Funds at beginning of the financial year
8,132,490
8,606,547
8,119,231
8,512,511
Adjustment to accumulated funds at the beginning of the financial year due to initial adoption of an Accounting Standard AASB1028, Accounting for Employee Entitlements.
14,137,023
8,060,027
14,033,825
7,956,072
2
Income tax attributable to Operating Profit
Aggregate of amounts transferred from reserves
3
(185,087)
(94,391)
257,550
257,550
The Company 1996 1995
$
$
$
397,450 1,125,685 2,205,028 3,728,163
331,571 715,917 826,808 1,874,296
375,749 371,547 2,036,439
234,684 328,210 562,535 2,783,7351,125,429
942,500 10,347,524 1,269,739 12,559,763
1,015,000 5,903,686 1,297,463 8,216,149
942,500 10,244,820 1,216,080 12,403,400
1,015,000 5,903,686 1,287,750 8,206,436
16,287,926 10,090,445
15,187,135
9,331,865
976,942 955,215 1,932,157
622,661 472,396 1,095,057
756,220 410,177 1,166,397
$
CURRENT ASSETS
Cash Operating Profit/(Loss) before Income Tax
Consolidated 1996 1995
Receivables Investments TOTAL CURRENT ASSETS NON-CURRENT ASSETS Receivables Investments Property, Plant &- Equipment TOTAL NON-CURRENT ASSETS
4 5
4 5 6
TOTAL ASSETS CURRENT LIABILITIES Creditors and Borrowings Provisions TOTAL CURRENT LIABILITIES
7 8
996,640 743,629 1,740,269
!
Total available for appropriation Aggregate of amounts transferred to reserves
14,137,023 8,132,490 3
83,319
Accumulated funds at the end of the financial year
14,053,704
14,033,825 8,119,231 83,319
8,132,490
13,950,506
NON-CURRENT LIABILITIES Creditors and Borrowings Provisions TOTAL NON-CURRENT LIABILITIES
53,859 162,874 216,733
49,600 165,444 215,044
56,240 56,240
43,595 43,595
TOTAL LIABILITIES
2,148,890
1,955,313
1,151,297
1,209,992
NET ASSETS
14,139,036
8,-135,132
14,035,838
8,121,873
8,1 19,231
MEMBERS’ FUNDS Accumulated Funds Reserves TOTAL MEMBERS' FUNDS The profit and loss accounts are to be read in conjunction with the attached notes to and forming part of the financial statements set out on pages 76 to 85. Page 73
7 8
3
14,053,704 8,132,490 2,642 85,332 14,139,036 8,135,132
13,950,506 8,119,231 85,332
2,642
14,035,838 8,121,873
The balance sheets are to be read in conjunction with the attached notes to and forming part of the financial statements set out on pages 76 to 85. Page 74
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THE MURDOCH INSTITUTE FOR RESEARCH INTO BIRTH DEFECTS LIMITED AND ITS CONTROLLED ENTITY
Notes to and Forming Part of the Financial Statements for the year ended 31 December 1996
Statements of Cash Flows for the year ended 31 December 1996 Consolidated 1996 1995 Inflows Inflows (Outflows) (Outflows) $
$
CASHFLOWS FROM OPERATING ACTIVITIES Payments to suppliers and employees (7,656,003) (6,400,451) Government Grants received 3,784,634 3,111,705 Donations received 7,037,446 1,149,756 Other receipts 1,079,844 629,712 Interest received 23,949 12,840 Patient fees received 765,311 662,326 NET CASH PROVIDED/(USED) BY OPERATING ACTIVITIES (Note 9(ii))
5,035,181
CASH FLOWS FROM INVESTING ACTIVITIES Interest received 722,83 1 Dividends received 85,504 Proceeds on sale of investments 3,289,742 Payment for investments (7,401,122) Payment for property, plant and equipment (192,353) NET CASH PROVIDED BY INVESTING ACTIVITIES
(3,495,398)
(834,112)
The Company
1996
1995
Inflows (Outflows)
Inflows (Outflows)
$
$
(4,306,857) (4,098,747) 1,374,434 1,431,905 7,037,446 1,149,756 955,145 648,680 10,978 2,886
5,071,146
(865,520)
263,404 705,611 249,615 84,109 85,504 84,109 7,315,112 3,025,468 6,906,112 (5,358,304) (7,129,829) (4,884,304) (1,218,705) (142,931) (1,218,705) 1,085,616 (3,456,177)
1,136,827
NET INCREASE IN CASH HELD
1,539,783
251,504
1,614,969
271,307
CASH AT THE BEGINNING OF THE REPORTING PERIOD
894,106
642,602
797,219
525,912
CASH AT THE END OF THE REPORTING PERIOD (Note 9(i))
2,433,889
894,106
2,412,188
797,219
The statements of cash flows are to be read in conjunction with the attached notes to and forming part of the financial statements set out on pages 76 to 85. Page 75
1.
STATEMENT OF SIGNIFICANT ACCOUNTING POLICIES
The significant policies which have been adopted in the preparation of these financial statements are;
(a)
Basis of Preparation
The financial statements have been drawn up in accordance with applicable Accounting Standards, the Corporations Law and Schedule 5 to the Corporations Regulations. They have been prepared on the basis of historical costs and do not take into account changing money values or, except where stated, current valuations of non'Current assets. The accounting policies have been consistently applied by the entities in the economic entity and are consistent with those of the previous year.
/
The carrying amounts of all non-current assets are reviewed at least annually to determine whether they are in excess of their recoverable amount at balance date. If the carrying amount of a non-current asset exceeds the recoverable amount, the asset is written down to the lower value. In assessing recoverable amounts the relevant cash flows have not been discounted to their present value. The financial statements of the Company and its controlled entity reflect all entries affecting the Company and its controlled entity and include transactions that were specifically recorded in the bank accounts of the Company and its controlled entity.
(b)
Principles of Consolidation
The consolidated financial statements combine the financial statements of the Murdoch Institute for Research into Birth Defects Limited, being the chief entity, and its controlled entity Victorian Clinical Genetics Services Limited, for the twelve months ended 31 December 1996. All balances and transactions between the chief entity and the controlled entity have been eliminated. Page 76
!■
(c)
twelve months are discounted using the rates attaching to national government securities at balance date, which most closely match the terms of maturity of the related liabilities.
Investments
Investments are stated at the lower of cost or recoverable amount. Dividends are brought to account as received.
(d)
In determining the liability for employee entitlements, consideration has been given to future increases in wage and salary rates, and the economic entity's experience with staff departures. Related on-costs have also been included in the liability.
Property, Plant and Equipment
Items of property, plant and equipment are recorded at cost. The depreciable amounts of all items of plant and equipment are depreciated over their estimated useful lives commencing from the time the asset is held ready for use. The straight line method of depreciation is used. Items of plant and equipment costing less than $5,000 are not capitalised. Leasehold improvements are amortised over 15 years - that being the same period of time as rent prepaid to the Royal Children's Hospital from the time that the laboratories and offices are ready for use.
(e)
(g)
Income Tax
The Company and its controlled entity are exempt from income tax under section 23(e) of the Income Tax Assessment Act 1936.
Jo
f
Prepayments
(h)
Revenue Recognition
Fee revenue comprises revenue earned from the provision of services to entities outside the economic entity. Fee revenue is recognised when the fee in respect of services provided is receivable. /
Items of expenditure having a benefit or relationship to more than one accounting period are amortised over the periods to which they relate.
(i)
(f)
Where necessary, comparative information has been reclassified to achieve consistency in disclosure with current financial amounts and other disclosures.
Employee Entitlements
Comparatives
Annual Leave
(j)
The provisions for employee entitlements to annual leave represents the amount which the economic entity has a present obligation to pay resulting from employees' services provided up to balance date. The provisions have been calculated at nominal amounts based on current wage and salary rates and includes related on-costs.
Research &- Development Expenditure
Research
Development Expenditure is expensed as incurred.
Long Service Leave
The liability for employee entitlements to long service leave represents the present value of the estimated future cash outflows to be made by the employer resulting from employees' services provided up to the balance date.
7
Liabilities for employee entitlements which are not expected to be settled within Page 77
Page 78
IV
Consolidated 1996 1995 $
The Company 1996 1995
$
$
Consolidated 1996 1995
$
2. OPERATING PROFIT
$
$
The Company 1995 1996 $
$
3. RESERVES
Operating Profit/(Loss) before Income tax has been determined after: (a) CREDITING AS REVENUE Grants ' NHMRC Grants ' H&-CS Grants ' Other Donations Interest' other persons Dividends ' other persons Net Gain on sale of investments Patient Fees
1,212,602 1,256,156 2,815,003 2,047,686 690,721 182,643 7,037,446 1,024,756 746,779 275,351 85,504 84,109 236,775 219,890 881,394 777,570
1,212,602 179,132 690,721 7,037,446 716,588 85,504 236,775
1,256,156 171,386 182,643 1,024,756 252,501 84,109 219,890
POSSUM Sales Income ' Other
119,653 380,056 14,205,933
129,431 240,564 6,238,156
119,653 283,809 10,562,230
129,431 294,331 3,615,203
Proceeds on sale of investments
3,289,742
7,315,112
3,025,468
6,906,112
(b) CHARGING AS EXPENSE SalariesAVages, incl. superannuation Employee entitlements Laboratory Consumables Refurbishment Costs Repairs &- Maintenance Travel Patient Care Services and Clinical Research Central Services &- Administration Depreciation POSSUM Costs Amortisation
5,153,655 4,461,622 400,165 280,185 822,037 576,714 214,862 1,403 148,203 37,098 123,827 79,283 400,229 322,523 589,538 752,628 148,195 141,806 128,807 131,414 71,882______ ^ 8,201,400 6,784,676
2,677,700 2,484,894 136,487 170,103 822,037 576,714 214,862 1,403 148,203 37,098 72,225 70,853 2,304
232,714 557,828 142,719 139,031 128,807 131,414 71,882 4,647,636 4,171,642
Total expenses incurred by the Murdoch Institute are for Research and Development.
Page 79
OI ASSOCIATION FUND SOCIAL WORK FUND Balance at end of year MOVEMENTS IN RESERVES SOCIAL WORK FUND Balance at beginning of year Transfer to profit &■ loss Balance at end of year BUILDING DEVELOPMENT FUND Balance at beginning of year Tansfer (to)/from profit &- loss Balance at end of year
83,319 2,013 85,332
2,642 (629) 2,013
2,642 2,642
83,319 2,013 85,332
2,642 2,642
2,692 (50) 2,642
2,642 (629) 2,013
2,692 (50) 2,642 257,500 (257,500)
257,500 (257,500)
OI ASSOCIATION FUND Balance at beginning of year 83,319 Transfer from profit &- loss 83,319 Balance at end of year The fund includes capital of $80,000 and interest there on.
83,319 83,319
4. RECEIVABLES CURRENT Patient fees due Less provision for doubtful debts
188,355 188,355
Amount Owing by Controlled Entity Debtors ' Sundry Prepayments Sundry Receivable NON-CURRENT Prepayments
864,830 72,500
130,844 (15,600) 115,244 513,511 82,162 5,000
114,647 167,776 184,400 73,272 72,500 82,162 -________5,000
1,125,685
715,917
371,547
328,210
942,500
1,015,000
942,500
1,015,000 Page 80
Consolidated 1996 1995 $ $
The Company 1996 1995 $ $
5. INVESTMENTS AT COST
1995 $
107,459 596,830 272,653 976,942
161,188 534,558 300,894 996,640
107,459 318,540 196,662 622,661
161,188 327,589 267,443 756,220
112,938 297,239 410,177 43,595
7. CREDITORS €r BORROWINGS
CURRENT Bank Bills and Deposits Commercial Bond Short Term Deposit TOTAL CURRENT INVESTMENTS NON-CURRENT Shares ' Listed on a prescribed stock exchange Debenture Stock Government Bonds - Unlisted Interest in Trusts TOTAL NON-CURRENT INVESTMENTS
1996 $
The Company 1996 1995 $ $
Consolidated
68,589 100,000 2,036,439 2,205,028
64,273 200,000 562,535 826,808
2,036,439 2,036,439
CURRENT Unearned Income Royal Children's Hospital Sundry Creditors
562,535 562,535
2,128,800 102,704
1,435,052
2,128,800
1,435,052
3,270,075 4,845,945 10,347,524
1,437,489 3,031,145 5,903,686
3,270,075 4,845,945 10,244,820
1,437,489 3,031,145 5,903,686
TOTAL INVESTMENTS
12,552,552
6,730,494
12,281,259
6,466,221
MARKET VALUE OF LISTED INVESTMENTS
2,677,135
1,795,965
2,677,135
1,795,965
TOTAL
NOTE: Royal Children's Hospital This is a suspense account which is used for payments to creditors. The Hospital pays the creditors on behalf of the Institute, and the Institute subsequently reimburses the Hospital. NON-CURRENT Special Purpose Funds
53,859
49,600
348,451 606,764
252,305 491,324
955,215
743,629
153,383 319,013 472,396
162,874
165,444
56,240
8. PROVISIONS
6. PROPERTY, PLANT
EQUIPMENT
Leasehold Improvements Accumulated amortisation Unamortised balance Plant &- Equipment - at cost Less: Accumulated Depreciation Written Down Value TOTAL
Page 81
CURRENT Annual Leave Long Service Leave
NON-CURRENT Long Service Leave
1,078,233 (71,882) 1,006,351 968,752 (705,364) 263,388 1,269,739
1,078,233 1,078,233 776,399 (557,169) 219,230 1,297,463
1,078,233 (71,882) 1,006,351 891,579 (681,850) 209,729 1,216,080
1,078,233 1,078,233 748,648 (539,131) 209,517 1,287,750
9. NOTES TO THE STATEMENTS OF CASH FLOWS (i)
RECONCILIATION OF CASH. For the purposes of the Statements of Cash Flows, the entity considers cash to include cash on hand and at bank and short term deposits at call. Cash as at the end of the reporting period as shown in the Statements of Cash Flows is reconciled to the related items in the Balance Sheets as follows: Page 82
Note
Consolidated 1996 1995 $
$
The Company 1996 1995 $
CASH
5
397,450 2,036,439 2,433,889
331,571 562,535 894,106
Add/(Less) Changes in Assets &- Liabilities (lncrease)/Decrease in Debtors ' Sundry (lncrease)/Decrease in Patient Fees Due (lncrease)/Decrease in Accrued Income (lncrease)/Decrease in Prepayments (lncrease)/Decrease in Amount owing and controlled entity (lncrease)/Decrease in other debtors lncrease/(Decrease) in Creditors lncrease/(Decrease) in Grants in Advance lncrease/(DecreaLse) in Accrued Expenses lncrease/(Decrease) in Unearned Income NET CASH PROVIDED/(USED) BY OPERATING ACTIVITIES
(236,775) (85,503) (722,830)
(219,890) (84,109) (263,404)
375,749 2,036,439 2,412,188
(236,775) (85,503) (705,612)
234,684 562.535 797,219
14,000
6,000
6,000
239,357
194,615
239,357
No.
No.
No.
No.
15
17
15
17
1
0
11. DIRECTORS’ REMUNERATION (556,439)
211,531 (69,771) (219,890) (84,109) (249,615)
(373,916) (73,111)
(97,539) (115,244) 125,000
(133,719)
64,162
82,162
2,174
82,162 53,129
125,000 5,696 (167,776)
5,035,181
$
Amounts received or due and receivable by the Auditors: 14,000 For auditing the accounts.
Directors' Income Total income received or receivable by the Directors of the Company from the Company or related bodies corporate.
33,341 4,363 66,570 6,529
31,739 (9,679) (17,300) (74,928) (36,429)
4,363 63,931 6,529
(834,112)
5,071,146
(865,520)
868
194,615
The number of Directors of the Company whose total income from the Company or related bodies corporate falls within the following bands: $0
31,739 65,901 (17,300) (32,388) (36,429)
$
10. REMUNERATION OF AUDITORS
(ii) RECONCILIATION OF NET CASH PROVIDED/(USED) BY OPERATING ACTIVITIES TO OPERATING PROFIT AFTER INCOME TAX. Operating Profit/(Loss) after Income Tax 6,004,533 (546,520) 5,914,594 Add/(Less) Non Cash Items Depreciation 6- Amortisation 220,077 214,306 214,601 Amounts set aside to Provisions 209,015 40,311 74,866 Add/(Less) Items classified ais Investing Activities Gain on sales of investments Dividends received Interest received
$
$
$
NOTES TO THE STATEMENT OF CASHFLOW CONTINUED
SHORT TERM DEPOSIT
The Company 1996 1995
Consolidated 1995 1996
$100,000 $120,000 $130,000 $190,000
^ ^ ^ ^
$9,999 $109,999 $129,999 $139,999 $199,999
0 0
The individual remuneration received the Directors in the band $0 ^ $9,999 was nil. Superannuation contributions paid in respect of directors are included in total income received The above amounts (including the comparatives) are disclosed in accordance with a ASC Class Order 96/1 171 dated 25/7/96.
12. LIABILITY OF MEMBERS In accordance with the Articles of Association, in the event of the company being wound up, members may be called upon to mahe a subscription not exceeding ten dollars. As at the 31 December 1996 the number of members of the company is 75.
Page 83
Page 84
r
THE MURDOCH INSTITUTE FOR RESEARCH INTO BIRTH DEFECTS LIMITED AND ITS CONTROLLED ENTITY
Stafemnent oy
13. RELATED PARTY DISCLOSURES
J^iFecfoFS
Transactions with Directors: Some directors made donations to the Company during the year. Transactions with the controlled entity: Financial assistance is provided by the Company to the controlled entity by way of infrastructure and overheads. At 31 December 1996 the company was owed by the VCGS $ 1 14,647. The names of each person holding the position of director of The Murdoch Institute for Research into Birth Defects Limited during the financial year: Professor J.A. Angus Mr. P. Griffin Professor G. Tregear Mr. L.G. Cox
Mr. J. Fitzgerald Professor P.D. Phelan Mrs. L. Cattermole Mrs. I. McFarling
Professor D.G. Penington Mrs. J. Calvert-Jones Mr. W.H. Hodgson Mr. I. Davies
Dr. G.L. Barnes Mr. J.S. Guest Professor R. Williamson Mrs. J. Paterson
Professor P.D. Phelan retired as a director on 27th March 1997.
/.
In the opinion of the Directors of The Murdoch Institute for Research into Birth Defects Limited:
(a)
the financial statements set out on pages 73 to 85 are drawn up so as to give a true and fair view of the results and cash flows for the financial year ended 31 December 1996, and the state of affairs at 31 December 1996, of the Company and the economic entity; (b) the consolidated accounts have been made out in accordance with Divisions 4A (c)
and 4B of Part 3.6 of the Corporations Law; and at the date of this statement, there are reasonable grounds to believe that the company will be able to pay its debts as and when they fall due.
2.
The financial statements have been made out in accordance with applicable Accounting Standards.
14. SEGMENT INFORMATION The Company and its controlled entity operates predominantly in a single industry, being research into causes, diagnosis and treatment of genetic diseases and other birth defects, within a single geographical segment, being Australia.
Dated at Melbourne this 23rd day ofApril 1997.
Signed in accordance with a resolution of the Directors:
15. PARTICULARS IN RELATION TO CONTROLLED ENTITY The Company is the only member of its controlled entity, Victorian Clinical Genetics Services Limited. No consideration was paid for this ownership, but in the event of the controlled entity being wound up, the Company may be called upon to pay an amount not exceeding ten dollars. The directors of the controlled entity are appointed by the Company.
LAURENCE G. COX Contribution to Consolidated Profit/(Loss)
The Murdoch Institute for Research into Birth Defects Ltd Controlled entity: Victorian Clinical Genetics Services Limited Page 85
1996
1995
$
$
5,914,594 89,939
(556,439) 9,919
6,004,533
(546,520)
; Directors
ROBERT WILLIAMSON
Page 86
INDEPENDENT AUDITORS’ REPORT TO THE MEMBERS OF THE MURDOCH INSTITUTE FOR RESEARCH INTO BIRTH DEFECTS LIMITED
Scope We have audited the financial statements of The Murdoch Institute for Research into Birth Defects Limited for the financial year ended 31 December 1996, consisting of the profit and loss accounts, balance sheets, statements of cash flows, accompanying notes, and statement by directors set out on pages 73 to 86. The financial statements comprise the accounts of the Company and the consolidated accounts of the economic entity, being the Company and its controlled entity. The Company's directors are responsible for the financial statements. We have conducted an inde pendent audit of these financial statements in order to express an opinion on them to the members of the Company. Our audit has been conducted in accordance with Australian Auditing Standards to provide reasonable assurance as to whether the financial statements are free of material misstatement. Our procedures included examination, on a test basis, of evidence supporting the amounts and other disclosures in the financial statements, and the evaluation of accounting policies and significant accounting estimates. These procedures have been undertaken to form an opinion as to whether, in all material respects, the financial statements are presented fairly in accordance with Accounting Standards and other mandatory professional reporting requirements (Urgent Issues Group Consensus Views) and statutory requirements so as to present a view which is consistent with our understanding of the Company's and the economic entity's financial position and the results of their operations, and their cashflows.
Corporations Law to be dealt with in the financial statements; (b) in accordance with the provisions of the Corporations Law; and (c)
in accordance with applicable Accounting Standards and other mandatory professional reporting requirements.
Dated at Melbourne this 23rd day of April 1997.
KPMG Chartered Accountants
R. Douglas - Partner
The audit opinion expressed in this report has been formed on the above basis.
Audit Opinion In our opinion, the financial statements of The Murdoch Institute for Research into Birth Defects Limited are properly drawn up: (a) so as to give a true and fair view of:
i)
ii) Page 87
the state of affairs of the Company and the economic entity at 31 December 1996 and the results and cash flows of the Company and the economic entity for the financial year ended on that date; and the other matters required by Divisions 4, 4A and 4B of Part 3.6 of the Page 88
MURDOCH ON THE WEB The Institute now has its own home page. at http://www.rch.unimelb. edu.au/murdoch/ A former Murdoch staff-member, Yvonne Harney, designed the page as part ofher computer studies. The page includes information about the Institute, staff vacancies, courses on offer, staffEmail addresses and details of the POSSUM/OSSUM database.
The Murdoch Institute for Research into Birth Defects Limited Royal Children's Hospital Remington Road PARKVILLE VICTORIA 3052 ACN 006 566 972 Postal Address: Post Office Box 1100 PARKVILLE 3052 Telephone: Facsimile:
(03) 9345 5045 (03) 9348 1391
Produced by Anne Cronin, Julie Foletta, Kati Hidegh, Michele Winsor Page 89
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Acknowledgements The Murdoch Institute for Research into Birth Defects Limited acknowledges the following donations:
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