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MURDOCH INSTITUTE

The incidence of birth handicap is 3 in 100 The cost in terms of personal and family [

SUFFERING IS ENORMOUS

The cost to Australia in terms of education and HEALTH PROVISION IS $2 BILLION PER ANNUM

The future lies in advances in genetics, for PREVENTION AND TREATMENT

Acknowledgements The Murdoch Institute for Research into Birth Defects Limited acknowledges the following donations:

mmssem Color scanning by Wilke Color.

Printing by Canberra Press

Canberra

OUR AIM IS TO HELP EVERY CHILD TO BE BORN HEALTHY AND WITH NORMAL ABILITIES

The Murdoch Institute IS AN INDEPENDENT RESEARCH ORGANISATION, COMPRISING I 80 DEDICATED STAFF DEVOTED TO ADVANCING KNOWLEDGE ABOUT GENETIC DISEASES

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Our Research

allenges

The structure and function of the human chromosome - how chromosomes divide in cells (for gene treat ment) and why this can go wrong (Down syndrome).

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■ To find the best ways to screen for inherited diseases.

The body's use of copper, and the genetic diseases causing copper deficiency (Menkes disease) and cop per toxicity (Wilson disease).

The understanding, diagnosis and treatment of con ditions affecting brain and muscle function in both childhood and adult life, including mitochondrial and metabolic disorders (such as phenylketonuria).

■ To conduct quality research of an international standard. ■ 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.

Understanding embryo development and how it con tributes to birth abnormalities (facial clefting, spina bifida and great vessel/cardiac defects) and cell dif ferentiation (paediatric cancers like neuroblastomas and leukaemias).

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. I

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 atax ias, cystic fibrosis, thalassaemia and other genetic disorders.

How Can You Help? You can help the Murdoch Institute reach its goal of helping every child to be born healthy through: ■

bequests

■

annual donations

The genetics of brain and nerve diseases: which mutations cause them, and developing new treat ments.

■

pledges over several years

■

named scholarships or research gifts

Discovering genes which cause deafness, blindness and other serious handicaps using Australian family resources.

For further information, please telephone or write to:

Our Staff We are a recognised centre of excellence in genetics and have attracted to our staff many world class doc tors, scientists, counsellors, nurses, administrators, and scholars.

Professor Bob Williamson Executive Director The Murdoch Institute Royal Children's Hospital Flemington Road, Parkville 3052 Telephone: (61) 3 9345 5045 Fax: (61) 3 9348 1391 Web site: http://murdoch.unimelb.rch.edu.au The Murdoch Institute is a registered charity. All donations over $2.00 are tax-deductible.

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Victorian Clinical Genetics Services

The MURDOCH^b^^TITUTI

Ac^evements and New Djrgctions

' *1®

Director

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Professor Bob t^nlia

Clinical Director

Victorian Clinical Genet^E^^jces . :

sfge 10

Chromosome Researbr^'

DrS.G. Kahler Trace Elements ;,v -

- -

i/ page 20

Gene Identification and Expn

Board of Directors

Mr. L.G. Cox, Chairman Mr M.J. O'D. Armstrong Dr G.L. Barnes Mrs J. Calvert-Jones Mrs L. Cattermole Dr J. De Campo Mr J.A. Fitzgerald

Mr L.G. Cox, Chairman Dr G.L. Barnes Professor H. Ekert Dr S.G. Kahler Mr W.H. Hodgson Mr G.E. Heeley Professor R. Smallwood Professor R. Williamson

Mr P. Griffin Mr W.H. Hodgson, Deputy Chairman Mrs A. McFarling Professor K. O'Dea Mrs J. Paterson Professor D.G. Penington Professor R. Smallwood Professor P.J. Smith Professor G.W. Tregear Professor R. Williamson

Gene Therapy

page 24

Gene Discovery

page 30

Metabolic Research Embryology

page 34 page 38

Disease Model Unit

page 42

Tissue Culture Laboratory Epidemiology POSSUM

page 44

page 46 page 50

Education

page 52

DNA Diagnostic Laboratory

page 56

Metabolic Laboratory

page 60

Cytogenetics Laboratory

page 62

Post-Graduate Diploma in Genetic Counselling

page 64

Maternal Serum Screening

page 66

Appreciation to our Supporters

Finance Committee 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 I. Miller Mr I. Veal

Operation Jigsaw Murdoch Institute Staff

page 70

page 74 page 78

Victorian Clinical Genetics Senrices Staff List of Publications

page 82

Financial Statements

page 92

page 80

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chievements and New

Directions

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Every organisation works to a vision and the Murdoch Institute is no exception. Our vision is to be one of the world's best human genetics centres across our entire range of activities. The result of our work in 1 997 has shown that, in this highly competitive field, we are performing extremely well and we have received wide international recogni tion. This comes in various forms, the high est being that our findings are being acknowledged by and are influencing the work of other scientists around the globe. We are not smug or complacent about our achievements or our reputation and appreci ate that they require enormous energy and innovation to be maintained and advanced. The Murdoch : Institute's strengths are the result of where yve are, what we are and who we are and it this knowledge which is fram ing our view of, what we can beconrip, ,

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Laurie Cox and Bob Williamson

Where we are

Who we are

is in a city and state which cares about medical research. We are in a precinct in Parkville which houses some of the world's best biologists and doc tors. Not far away are excellent universities with whom we collaborate. We share our building with one of the world's great children's hospitals. Our location is an important factor in our continued development and we plan to put considerable effort in the coming year into strengthening our links with this impressive network. One tangible example of that has been our attempt to encourage Government support for research.

is a group of dedicated researchers, who want to be tuned into the needs of the Australian community and economy.

What we are is the envy of many because we are not a stand-alone biomedical institute. We also care for children, adults and their families. We learn from the commu nity and are able to study questions which are highly relevant to the bedside and the clinic. The Victorian Department “of.. reaffirmed its support for the concept of the Victorian Clinical Genetics Service (VCGS) as a model State wide Genetics Service and given us initiative-led bud get ingfeases, which have allowed the VpGSjl) con-

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tinue to grow both in size and scope. We work with twenty hospitals throughout Victoria, are on call 24 hours a day and are expanding to work collaboratively in areas of adult medicine. We have been able to employ new training Fellows, specialist clinicians and genetic counsellors. All of these represent a superb foundation for the new VCGS Clinical Director, Professor Stephen Kahler, who arrived from Duke University in the United States in early 1998.

For example, the Cooperative Research Centre (CRC) for Human Gene Discovery is under way. We are participating in a partnership of industry and researchers which aims to translate theoretical dis coveries into uses and products. Through the CRC, which also involves AMRAD, we have been searching in collaboration with the Royal Children's Hospital Allergy Unit for genes which predispose to eczema. Eczema causes untold suffering to babies and their families and is hard to treat. If we find predisposing genes, we could then attempt to prevent the onset of eczema through attention to breastfeeding and aller gen avoidance. If we succeed, we hope that start-up biotechnology groups in Victoria, using financial incentives established by successive State and Federal governments, will enable us to develop the discover ies in Australia, rather than watch them go offshore. The scientific teams within the Murdoch have done well during 1997. Andy Choo's group has succeed ed in causing a "paradigm shiftTiB our understanding of the human chromosome. In q muph-lauded Nature Genetics paper he described a new way that the body controls how chromosomes pass themselves onto their ddughtbr cells. To'illustrate the importance

of our clinical links, thislincTing is Sase^a bn the enro^ mosomes of a patient seen by our medical team at the Royal Children's Hospital.

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Other research which is steadily bearing fruit includel^- , work on how cells migrate during very early embryr' onic development. Many of the birth handicaps* which we see and treat arise from errors at these early* stages of life. * We are developing new ways to look at genes which predispose to complex diseases such as cleft palate and allergy, and we are searching for genetic causes of deafness. Previous studies on deafness suggested that many different genes are involved but in fact one mistake in one gene seems to be the cause of deaf ness in about a third of the cases which occur in Victoria. The work of our epidemiology unit continues apace and they have started several studies to determine "who, how and why" people use our Victorian genet ic services. We are also making an impact on gene therapy research. Our clinicians produce a steady stream of papers, and two major books (one on the centromere, one on cytogenetics) were published from the Murdoch this year. In education, we now have a committed unit which offers courses to all from primary schoolchildren to senior clinicians, and runs Operation Jigsaw, an edu cation program for Victoria's schools which also rais es funds for our research. Julian Savulescu, a very distinguished medical ethicist, has been appointed to head our unit to study the Ethics of the New Genetics, with close links to the University of Melbourne Centre for the Study of Health and Society. The best research in the world is of little use if the community cannot understand it or suspects it may not be ethical. We intend to devote time and resources to ensuring that both education and ethics receive due attention in Victoria.

Whereiare we going? I?

Government funding of NH&MRC

To succeQcl in the next millennium, we must recruit the very bestptudents, post-docs, clinicians and slipport staff to the Murdoch. This will require imaginat ve use of space! and resources.^? We are grateful to the Department of Human Services and our benefpctors, for providing the'money to rebuild our cliniea wing. This pro\|des a state-of-the-art facility with aroper counsellirig and clinical areas and all the information links and laboratory facilities a genetics service requires. We are finding ways to increase our collaboration with other teams, locally, nationally and internation ally and look forward with great optimism to meeting the challenges ahead for both the Murdoch and the Victorian Clinical Genetics Service. The staff of the Institute is well equipped to provide the muscle to drive advances forward in many areas of human genetic research and to ensure they can be used in the interests of all Australians Laurie Cox and Bob Williamson

In MiD-1997y it became clear that NH&MRC funding COULD BE CUT BY UP TO 25% OVER THE THREE YEARS TO 2000. If this happened, there would be no money for new bio

ANOTHER CAREER, WORKING FOR EXCELLENCE IN BIOMEDICAL RESEARCH IN VICTORIA IN MANY CAPACITIES, BUT PARTICULARLY THROUGH HIS

RESEARCH, AND MANY YOUNG AUSTRALIAN DOCTORS AND SCIENTISTS WOULD BE FORCED ABROAD TO FOLLOW A RESEARCH CAREER. WiTH THIS IN MIND, YOUR BOARD AND SENIOR STAFF HAVE ENCOURAGED THE GOVERNMENT TO LOOK AGAIN AT

STEWARDSHIP OF THE BrOCKHOFF FOUNDATION. He IS A TESTIMONY TO THE FACT THAT AT LEAST

FUNDING FOR MEDICAL RESEARCH. IN A FLOURISHING LANDSCAPE WHICH INCLUDES THE UNIVERSITIES AND GOVERNMENT

SOME 80-year-olds can still run AROUND MOST OF US INTELLECTUALLY.

INTERNATIONAL INDICES, EXTREMELY SUCCESSFUL IN RESEARCH.

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PRACTICE

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medical

Departments, the biomedical Institutes are, as judged by

rings

Ivor Davies retired as CEO of the Women's AND Children's Health Care Network. Ivor

We hope that our efforts to encourage Government sup port FOR RESEARCH, COORDINATED WITH THOSE OF COLLEAGUES

WAS A GREAT HELP IN MEETING THE CHALLENGES OF THE MANY CHANGES IN HEALTH CARE IN Victoria, was a loyal friend to the Institute

IN OTHER Institutes, will succeed at least to the extent that FUNDING FOR NH&MRC WILL BE MAINTAINED DURING 1 998. A

Ministerial Review of Health and Medical Research has NOW commenced under Peter Wills, the Chairman of the Board of the Garvan Institute, which we welcome and we

AND WORKED WITH US TO FORGE CLOSE LINKS WITH THE Network.

OF WISDOM REQUEST.

Julian Mercer Julian Mercer's work on copper has been of During the past year,

' Julian HAS begun to study environmental ASPECTS OF COPPER METABOLISM IN ADDITION TO HIS LONG-ESTABLISHED STUDIES OF MeNKES AND Wilson DISEASES. Julian has decided to move TO Deakin University to take up a prestigious POST AS Director and Professor of Cell and Molecular Biology. During the past twen ty YEARS, Julian has played a key role in the scientific track record of the Murdoch, PARTICULARLY THROUGH HIS SEMINAL WORK WITH

David Danks on Menkes disease.

HOPE WILL LEAD TO NEW GOVERNMENT INITIATIVES WHICH IMPROVE

Jim Angus, a very distinguished pharmacol ogist, HAS represented THE NH&MRC ON THE Board. He has been a constant source

THE HIGHEST QUALITY.

' Three members of the Murdoch Institute Board have retired during 1997, James Guest, Jim Angus and Ivor Davies. James Guest is one of the most distinguished surGEONS OF HIS GENERATION. On his retire-

AND

HELP,

GIVEN

FREELY

While we will miss their presence around the Board table, we wish each of them many MORE PRODUCTIVE YEARS.

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One measure of the excellence of a country - the reason WE BELIEVE WE DESERVE "FIRST WORLD STATUS" - IS THAT WE CARE ABOUT THE HEALTH OF OUR PEOPLE. A PART OF THAT MUST BE CONTRIBUTING TO HEALTH CARE RESEARCH AT THE HIGHEST LEVEL.

First world countries do not live off the backs of oth ers,

BUT PLAY THEIR PART. THUS WE ARGUE THAT THE AUSTRALIAN

GOVERNMENT HAS A RESPONSIBILITY TO ENSURE A HEALTHY BIO MEDICAL RESEARCH COMMUNITY THROUGH THE NH&MRC.

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We will

CONTINUE TO COLLABORATE WITH JULIAN DURING THE COMING YEARS, AND WISH HIM WELL IN HIS NEW POSITION.

STILL FURTHER THE COMPETITIVENESS OF AUSTRALIAN BIOMEDICAL RESEARCH.

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In this context, it is worth noting that President Clinton AND HIS Congress speak of doubling the NIH budget for MEDICAL research OVER FIVE YEARS, WITH AN INCREASE OF $2 BILLION (U.S.) BASED ON A STARTING BUDGET OF ABOUT $15 BILLION IN 1999. Our total NH&MRC budget is less than A hundredth of the current American NIH research spend!

[ We also welcomed Dr Avihu Boneh in 1997, who joined us to provide clinical care to children with metabolic disease. These young people can often be helped dramatically by diet and new therapies, and we have a strong support team including genetic nurses, a social worker and a dietitian providing resources to the families. Many of those who previ ously died early in childhood are now living more or less normally into adult years, which raises new issues for the service, as we have to provide clinical care through adult hospitals and not only at the Royal Children's Hospital and other paediatric centres.

he 1/ictbrian| m Clinical I Genetice^ ■ t si Services' .1/3

The Victorian Clinical Genetics Services (VCGS) offers clinical genetics care, and genetic counselling, and gene analysis, to the whole population of Victoria and Tasmania. It is closely integrated with the research teams, each facilitating the work of the other. Clinical geneticists, genetic coun sellors and diagnostic scientists all receive their training through the VCGS; our staff and laboratories are fully accredited to the highest standards. We see the provision of this service to all Victorians, with continuous consideration of ethical aspects, and always reviewing their need for help and informa tion, support and treatment, as the driving force in all we do.

1997 was a year of growth and develop ment for the VCGS, although the single most important change took place at the start of 1998 - the arrival of Dr Stephen Kahler to become the new Clinical Director. With the full support of the entire VCGS and Murdoch team, we expect that Steve will be able to continue to develop the service, building on his knowledge developed in Duke University, one of the best of the United States medical centres.

During 1997, the VCGS was assessed by a team of internationally recognised experts in clinical genetics Peter Harper (University of Wales), Eric Haan (Adelaide) and Bridget Wiicken (Sydney) joined us for two days and examined every aspect of the work of the Service. Their report, which was to our Board but also welcomed by the Department of Human Services, affirmed that we really live up to the stan dards to which we aspire, and their complements were gratifying to all staff. However, they also point ed to some areas where changes could be made - for instance, they encouraged a transition to greater shared involvement in adult services through Monash Medical Centre, the Royal Melbourne Hospital, the Alfred and Austin, St Vincent's and Peter MacCallum Hospitals.

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They also noted the need for particular emphasis on cancer genetics services, and for outreach clinics and counsellors based in country Victoria. All of this has to be carried out while continuing to provide a first rate service in a paediatric context at the Royal Children's Hospital, where many of our clinics are held, and Monash Medical Centre, as well as inpa tient care for severely ill children with metabolic dis eases. We know that these recommendations will strike a positive note with everyone reading this report, and we hope that the Minister will also see our efforts deserve support, since increased services have to be backed by a budget. Although we have been able to start several new clinics, and offer more clinical care and counselling than before, this has meant that vir tually all staff are stretched to the limit, and there is too little time for audit, research and initiative. Hub and spoke services such as ours require adequate numbers of first rate staff at both hub and spoke, if the service is to work well.

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The educational activities of the VCGS have contin ued to multiply - our Diploma course for Genetic Counsellors was a great success, with all ten students qualifying with excellent grades. We are particularly pleased that some of those who have gone through the program are using their skills to improve their cur rent job performance and satisfaction, working with cancer or Alzheimer disease patients or with handi capped adults. It is interesting that the biggest increase in use of our service, by about 50% in one year, has been for genetic counselling. It is not enough to provide tests for genes that are mutated; the families that are affected have to understand the consequences and work through their own solutions which are consistent with their fundamental beliefs.

Of the senior staff, we are very pleased that Agnes Bankier has been chosen to be President-Elect of the Human Genetics Society of Australasia, the accredit ed professional body to which all clinical geneticists, counsellors and diagnosticians belong. This is a sig nal honour for Agnes, and of course we applaud her success. Les Sheffield is spending a year at Yale and NIH working on analysis of complex human genetic diseases, which is one of the hottest topics in human genetics today, and Mac Gardner is playing a major role in research into genetic causes of deafness, as well as overseeing the publishing of the 2nd edition of his book with Grant Sutherland, "Chromosomal Abnormalities and Genetic Counselling".

Of our five genetic Fellows, all did well, and three will return during 1998 to continue their training and be joined by three further colleagues. Ravi Savarirayan achieved the remarkable honour of being chosen to be the Fulbright Fellow for Australia for 1998, the only one in the professional category. He will study skeletal dysplasias in California at Cedars-Sinai Hospital. Simon Hauser will return to specialty pae diatrics but will continue to work closely with the VCGS, and Karen Dunn has gone to set up paediatric services in Goradze in Bosnia, working for a medical charity for a year. Our visitors have also done well Francois Bernier has been appointed to a senior post in Montreal, Canada and Helen Heussler has obtained a research post in Birmingham, U.K. Of course, our educational activities are also interna tional - the world renowned POSSUM computerised data base system to help clinicians to identify dys morphic syndromes is now in use in over 50 coun tries, and is widely regarded as the first and still the best of its kind.

There have been several new initiatives. The VCGS has been able to provide ongoing support for the introduction of maternal serum screening using four markers. This test gives all pregnant women a choice of receiving information about possible risks of carry ing a fetus with a neural tube defect such as spina bifida, or Down Syndrome. We are particularly pleased that funding has been provided to allow us to offer the screening test to all women, whether public or private patients. We have also been supported to improve the testing of families which have several cases of colorectal cancer, and the mutations in one form of the disease (familial adenomatous polyposis, or FAP) have now been identified in almost every fam ily affected in Victoria.

Not only is this a "world first", it even saves money, since family members who do not inherit the mutation do not need a costly invasive test every six months. During 1 998, we will look forward to the opening of the new Clinical Wing. This development has been part funded by the Department of Human Services, part funded by generous donations in honour of Dame Elisabeth and her role in supporting the clini cal genetic service and genetics research in Victoria. The VCGS will be able to make full use of the new premises, for the future demands on clinical genetics by the community will increase, and will require all of our effort, commitment and the highest standards over the coming years.

Agnes Bankier (Clinical Director, 1997) Bob Williamson (Executive Director)

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Understanding how chromosomes divide A milestone of the Chromosome Research Group's research this year has been our breakthrough in understanding the nature of the human centromere. Our discovery received national newspaper coverage when our paper entitled "A functional neo-centromere formed through activation of a latent human centromere and consisting of non-alpha-satellite DNA" was published in "Nature Genetics". Every time cells divide a new copy of each chromosome is made. The cehfrorfiere is the part of ,a chromosome which pffovides the machinery to distribute new chrorrlbsome copies equfally between the two newly formed cellsf In the past, it was alwaW held that the hurnan centromere was madp up of a single type of repetitive DNA, l|ut our research has now shown that this is npt true.

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We discovered that many different types of DNA can become a centromere. This is an exciting new discovery which should acceler ate our understanding of how the chromo some divides. We are currently studying this DNA in detail and using it to construct human artificial chromosomes for use in gene therapy.

Centromere protein mutations We have also made important advances in learning what some of the centromere proteins do. We have mutated a number of genes that make these proteins in laboratory mice. One mutated gene causes a cat astrophic misdistribution of chromosomes during cell division and leads to early fetal cell death during pregnpncW'""Wce''cari7i^.a'secbnd..rn'utate^ gene look |ormal and reproduce well, but are lofver in body weight, testes weight and sperm content. These, and other mice carrying new mutations, will be stud ied in detail.

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Defining a new paradigm on the dynamic nature of centromere DNA

Centromere pi Ptudy^ by gene knockout

The past four to five years of our laboratory work has concentrated on the study of an unusual centromere of a human chromosome 10-derived marker chro mosome that lacks the large amount of repetitive alpha-satellite DNA found in normal centromeres. By employing chromosome walking using fluorescence in situ hybridisation (FISff) and anti-centromere anti body staining, we have now narrowed the functional ly critical region to less than 80 kb and have cloned this DNA. Direct sequencing of this DNA has revealed no resemblance to the normal centromere DNA. Restriction mapping comparison of the mark er centromere DNA with the corresponding region of the normal chromosome 10 where the new cen tromere (or neocentromere) has formed has demon strated no major difference between the two DNAs, suggesting that the neocentromere has originated from the activation of a DNA sequence with the prop erties of a latent centromere.

The study of centromere: proteins complements: our ; ; DNA wofc and is an important componenf of bur centromere programme. One of our approaches T A involves " :r(©cMng oat" o Gentromere-bmding protein ^ in transg inic mice: and: studying the effects of the " mutation. .We fiypothesise that .mutations in most, if „ not all, oF the TncreasmgnTi7mEerO)in<nown~ceh-" tromere-binding proteins are a major cause of chro mosome mis-division and aneuploidy that can lead both to fetal loss during pregnancy and to Down syn drome. We have now produced null mutations (by homozygous gene knockout) for a number of these proteins. Deficiency in one of these proteins, cen tromere protein C, results in early embryo death due to severe mitotic disarray and extensive micronuclei formation, providing support for our hypothesis. Deficiency in a second protein, however, yielded a different outcome in that the mice appeared to be healthy and reproductively normal, although these animals have reduced body and testes weight, and lower sperm numbers. The underlying reason for these reductions, as well as the effects of knocking out other centromere proteins, are currently being inves tigated.

Our study provides the first molecular evidence for a new genetic paradigm. This paradigm stipulates the existence of latent centromere sequences that can be activated to from neocentromeres in the human genome. It also brings to light the dynamic nature of the centromere DNA, in that seemingly unrelated DNA sequences can become centromeres. This par adigm appears to be widespread throughout evolu tion since it has now also been demonstrated in Drosophila, a fruit fly often used for genetic studies due to their ease and rapidity of breeding. Our cur rent work is aimed at understanding the nature of our neocentromere DNA, its functional elements and mechanisms of silencing and activation, and its appli cation in the synthesis of human artificial chromo somes as novel vectors for carrying corrective genes for patient treatment in human gene therapy.

Andy Choo Liz Earle Linda Hii Dani Irvine Kellie Tainton Ben Kile Ainsley Newson Alyssa Barry Emily ffowman Damien Hudson Paul Kalitsis Anthony Lo Andrew MacDonald Michael Concilia Suzi Cutts Richard Saffery

Group Leader Scientific Officer Research Assistant Research Assistant Research Assistant BSc Honours Student BSc Honours Student PhD Scholar PhD Scholar PhD Scholar PhD Scholar PhD Scholar PhD Scholar Postdoctoral Scientist Postdoctoral Scientist Postdoctoral Scientist

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race Elements Group

The Trace Element Group studies how the essential trace element, copper, is carried around the body and within cells. Although copper is essential for life, since it is needed for many important enzymes, it is also poten tially toxic so its transport must be carefully controlled to avoid damage to cells. Our particular interest is to determine the molec ular basis of the genetic diseases which dis rupt this process of copper transport. The two main diseases we study are Menkes and Wilson diseases. Menkes disease is a genet ic copper deficiency, which leads to death in early childhood. Wilson disease is a copper toxicosis condition resulting in liver failure or brain damage and death in the teenage years unless treated.

Michael Petris, Sharon La Fontaine and Julian Mercer

Over the last few years we have achieved a major breakthrough with the isolation of the gene affected in Menkes disease, which 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 causing Menkes disease in the affected families, 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, which not only allows us to study the normal function of the gene, but also should lead ultimately to gene therapy of Menkes and Wilson diseases.

ma We continue our work towards understanding how cells regulate copper. In collaboration with Dr. Jim Camakaris's laboratory (University of Melbourne), we have been investigating the intracellular location of the Menkes protein (MNK) in Chinese hamster ovary cells. We have found 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, which 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 was published in EMBO J in 1996 A major focus of our research is to determine the mol ecular basis of this copper-regulated MNK move ment. It is important to investigate this process because we believe it to be the basis of copper home ostasis at a cellular level. It is a novel mechanism in cell biology, and it will help to explain the clinical fea tures of Menkes disease. Significant progress has been made in obtaining sta ble plasmid constructs containing the protein coding region of the Menkes gene. When introduced into CHO-Kl cells, these plasmids produced Menkes pro tein (MNK) which was functional in both copperinduced trafficking between the transGolgi network (TGN) and plasma membrane, and in conferring a copper resistance phenotype upon the cells.

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These studies represent the first demonstration of the successful expression of this protein from a cDNA construct in mammalian cells. In addition, using the CHO cell lines that we have generated, ultrastructural studies have shown the association of the Menkes protein with the TGN, plasma membrane and vesicles within the cell. N-terminal mutagenesis of the Menkes protein is in progress by Daniel Strausak and Sharon La Fontaine to determine the role of the N-terminal metal binding region of the protein in the copper-induced trafficking of the protein. Preliminary results show that mutation of the critical cysteine residues that are part of each metal binding region in 3 or 6 metal binding sites abolishes the copper-induced redistribution of the protein. Other studies in progress include the introduction of MNK plasmid constructs into cells from Menkes patients to determine if the copper transport defect in these cells can be corrected. Clones that have taken up the DNA and express the protein are currently being isolated and purified. The Mottled mouse mutants represent convenient models for Menkes disease. Mutations in the blotchy and brindled mice, which are analogous to mild and more severe forms of the human condition, respec tively, have been characterized.

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We have been able to detect the Menkes protein in cells from the brindled and blotchy mouse by immunofluorescence analysis, but it appears that in these cells the protein does not traffick in response to copper. It is possible that inactive Menkes protein in these cells is unable to participate in the protein inter actions that lead to the copper-induced movement of MNK. Plasmid constructs containing the protein coding region of the Wilson gene, and that of the mutated form of the gene from the toxic milk mouse (tx), a model of Wilson disease, have been generated. These constructs have been introduced into CHO-Kl cells to generate stable cell lines expressing the nor mal and mutated form of the Wilson protein (WND). These cell lines will be characterised to determine where the WND; protein is located within the cell, whether it trafficks in response to copper in a similar manner to the Menkes protein, whether it cdn confer a copper resistance phenotype upon the cells, and the affect of the tx mutation on the function of the WND protein.

We are continuing to analyse genes from some of our Menkes patients to identify mutations. Initially work is focussed on one patient who has been treated suc cessfully with copper and another patient with mild Menkes disease. We have also succeeded in detect ing the Menkes protein in fibroblast extracts using Western blots. In most Menkes patients, no MNK protein was observed, but in the mild Menkes and the copper treated patient MNK was detected 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. Work is continuing on animal models of Wilson dis ease including sheep and mice. The normal sheep can accumulate dangerous levels of copper in the liver if the feed supplements contain too much copper and in this respect resembles patients with Wilson dis ease. To investigate this possibility we have isolated and sequenced cDNA clones of the sheep Wilson homologue. No obvious mutations have been found which would indicate the protein was inactive. However, we did find a novel form of the WD protein with an extended N-terminal sequence which may play d'TOleln fhe’uhusudT copper horfiiosfoilf in the sheep. We are developing functional assays tc|determine whether the normal or extended form of t|e pro tein has copper transport activity, and whether each is localised correctly in the cell.

The mouse model for Wilson disease (toxic milk mouse of Wilson disease tx ) also accumulates cop per to very high levels in the liver, like 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 methio nine in the copper channel through the membrane (Theophilos et al, 1996). This proves that the mouse is a true model of Wilson disease, and opens the way to more extensive use of this mouse for studies on treatment of this disease. We are investigating the effect of copper loading the tx mouse to see if this precipitates a more acute form of the disease. A model of the acute form of the disease will allow us to investigate treatment strategies. Dr Katie Allen, a paediatric gastroenterologist carry ing out research for her PhD with the trace elements group in collaboration with the gene therapy team, has also made progress towards investigating gene correction of Wilson disease. We have established procedures of isolating mouse hepatocytes and re introducing them into the mutant mouse. We have also derived a temperature-sensitive conditionally immortalised liver cell line for in vitro gene correction with the Wilson disease cDNA construct. Work is in progress to correct the tx mouse phenotype (the mouse model for Wilson disease) using hepato cellular transplantation of non-mutant haplotypes. These experiments will pave the way for future ex vivo gene therapy for the tx mouse using gene-corrected haplotypes.

Tfm Kumaran Narayanan is also collaborating with the gene therapy group to use the mouse model for Menkes disease as a target for neuronal gene thera py. Although copper deficit in Menkes patients can sometimes be treated in most major body systems, the brain still becomes damaged because copper cannot transverse the blood-brain barrier. We are attempt ing to prepare chromosomal constructs which can target neurons and introduce the Menkes gene, with the aim of introducing a functional copy of the gene- j *3 into the choroid plexus to allow transport of copper into the CSF. Although Julian Mercer's move to Deakin early in 1998 as Professor of Molecular and Cell Biology will see the relocation of the Trace Element Group at the end of 1998, ongoing collaboration, in particular with Katie Allen and Kumaran Narayanan in the gene therapy group will provide an ongoing link with the trace metal team.

Julian F.B. Mercer Sharon La Fontaine Daniel Strausak

Andrew Grimes Stephen Firth Jenny Paynter Katie Allen Loreta Ambrosini Michelle Howie Paul Lockhart Michael Petris Michael Theophilos Rosario Reyes Jim Camakaris

Group Leader Postdoctoral Fellow Postdoctoral Fellow (1 997 fellowship funding from Hoffman La Roche) Research Officer Research Assistant Research Assistant (to March 1997) Clinical PhD Student PhD Student PhD Student PhD Student PhD Student Helen M. Schutt, PhD Student Technical Assistant Senior Associate of University of Melbourne

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Proper mitochondrial function is essential to generate energy in forms that can be used to drive cellular reactions. Mitochondria are also central in maintain ing a correct intracellular environment. The functions of these complex organelles are dependent on the interaction of a large number of gene products from both the mitochondrial and nuclear genomes. Defects in mitochondrial function are relatively com mon, but the underlying causes are often complex or unknown. As a consequence, both treatment and counselling are, as yet, unsatisfactory.

Both the understanding of a genetic disease, and helping those affected by the disorder, often requires detailed knowledge about which genes are affected and the way they are expressed. A typical exannple of this is in inherited deafness. A nnajor focus of our work in the past year has been on genes causing hearing loss. We have found a genetic mutation which seems to be a very common cause of childhood deafness, and this work has opened up new and very excit ing avenues which will benefit affected families. Henrik Dahl

Inherited Childhood Deafness

We have continued to study mitochondrial disorders, with special focus on identifying nuclear genes causing respiratory chain defects and on analysing how mitochondrial DNA is inherited in mice. Another project aimed at identifying a gene causing atten tion deficit hyperactivity disorder is also pro gressing well. Finally, we have found muta tions in families with chondrodysplasia punc tata and Bethlem myopathy, results that have provided new and, in some cases, unexpect ed information on these disorders.

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Finally, in collaboration with Dr. Jeff Mann (U.S.A.) and the Mouse Model Unit, we have made mice with two different mitochondrial DNAs. These mice are proving useful in studying the special way in which mitochondrial DNA is inherited and distributed in var ious tissues.

The incidence of prelingual deafness (at birth or very early in childhood) is approximately 1/1000 births, of which more than half are due to genetic causes. The development and proper function of the ear is com plex and requires the correct and co-ordinated expression of many genes. It is therefore not surpris ing that deafness can be caused by changes in a wide range of genes. One such gene is DFNBl. Defects in this gene are the most common cause of recessive, non-syndromic deafness - mutations in this gene may be responsible for deafness in 1/4000 children. Recently a group from England reported the identifi cation of the DFNBl gene. Our team at the Murdoch Institute has been involved in the characterisation of mutations of the DFNBl gene. This gene codes for a protein (connexin 26) involved in transporting small regulatory molecules and salts in and out of cells in the inner ear. Mutations in the DFNBl protein affect the function of the cochlea. We discovered that most people with recessive hearing loss have a common mutation in the DFNBl gene. With the new knowledge of the molecular and func tional details of the DFNBl gene it is now possible to begin a genetic approach to diagnosis and treatment of deafness. For example, there might be implica tions with respect to the success of cochlear implants in children.

We have continued the molecular investigation of res piratory chain disorders in close collaboration with David Thorburn and the Metabolic Disease Group. Using linkage and chromosome analysis we are hom ing in on new nuclear genes responsible for complex I and IV deficiencies. It is especially difficult to predict the severity of disease caused by mutations in the mitochondrial DNA. We are completing the first study in which the mutant load in the mother is corre lated with the severity of the disease in the child. These data are particularly important in genetic coun selling and predicting the recurrence risks.

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symptoms of the disease can transmit the mutations. Characterisation of these mutations allows further analysis of the phenotype, prognosis and molecular pathology, as well as providing essential information for counselling of affected families. Patients with Binder syndrome have facial hypoplasia, including a short nose with a flat bridge. Plastic sur geons are very familiar with this syndrome as many patients seek medical help during adolescence. Dr. Sheffield has suggested that Binder syndrome is a mild form of chondrodysplasia punctata. We have therefore initiated a search for mutations in the arylsulfatase genes in patients with Binder syndrome.

Chondrodysplasia Punctata and Binder Syndrome

Bethlem Myopathy

In collaboration with Les Sheffield (VCGS) we hae continued the study of patients with chondrodysplasia punctata. This is a genetic syndrome characterised by skeletal abnormalities, growth disturbance and facial dysmorphism. This disorder can be caused by mutations in arylsulfatase genes on the X chromo some. We have identified individual mutations in the arylsulfatase E gene in 4 families, have shown segre gation of ARS-E mutations with the disease, and pre sented evidence that females and males without

Muscular dystrophies are a heterogeneous group of diseases characterised by progressive muscle weak ness and wasting. We have identified a new patho genic mechanism for an inherited muscular dystrophy in which having only half of the normal amount of the protein collagen VI causes Bethleffi myopathy.!These exciting data are the result of a collaboration with Dr Shireen Lamande (Orthopaedic Research) Unit, R.C.H.), Mac Gardner (VCGS) and Prof. Ed Byrne's group!(Melbourne Neuromuscular Researchjjentre.

Attention Deficit Hyperactivity Disorder In collaboration with Sue Forrest and Martin Delatycki we have initiated a study aimed at identifying a gene for attention deficit hyperactivity disorder (ADHD). As many as 5-10% of school children are showing symp toms of ADHD, affecting their education and social development. It is clear that genetic factors play a significant role in this disorder. However, so far no genes been shown to cause ADHD.

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St. Vincent's Hospital). Data were obtained by study ing a family with Bethlem myopathy. The defect in this family was mapped to the collagen 6A1 and 6A2 genes on chromosome 21q22.3 and shown to be a single base deletion in an intron of the collagen 6A1 gene. The mutation moves the splice junction one base, resulting in a frameshift in the mRNA. Theoretically, this should result in a truncated 37 kDa collagen 6A1 -like product. However, the mutant mRNA is unstable and decays rapidly, and is almost completely absent from patient fibroblasts (skin cells), resulting in lack of the collagen 6A1 subunit and reduced production of structurally normal collagen VI. This is the first example of a muscular dystrophy caused by haploinsuffidency (having only half the normal amount) of a structural protein, and identifies collagen VI as a critical contributor to cell-matrix adhesion in skeletal muscle.

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A family has been identified in which some members have behavioural problems with features of attention deficit hyperactivity disorder. Co-segregating with this phenotype is a chromosome abnormality. So far 11 members with the chromosome change have been found, and we are assessing family members with and without the chromosome rearrangement by formal neuropsychological testing. Fibroblast and lymphoblast cultures have been set up from two carriers of the rearrangement aiming to iso late clones from the breakpoints and to identify a gene responsible for the observed phenotype. Fluorescent in situ hybridisation (FISH) technology using YAC (yeast artificial chromosomes), PAC (PI artificial chromosomes) and cosmid clones is being used to identify the chromosome breakpoints. Once identified we will look for mutations in this ADHD gene to study genotype/phenotype correlations and also determine whether mutations in this gene are a common cause of ADHD.

Henrik Dahl Stephen Wilcox Wendy Hutchison Amelia Osborn Laraine Peters Sarah White Carly Selan Marietta Veldman

Group Leader Postdoctoral Fellow Scientific Officer Research Assistant PhD Student PhD Student Honours Student Visiting Medical Student

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Mark Twain, the American novelist, is reput ed to have said "I have just found out that I have been speaking prose all my life, and I never knowed it". I feel a bit the same about "gene therapy". Gene therapy is the study of how to introduce a gene into the cells in the body of a person who is not well, and express it to treat disease. For geneticists, it is only a small extension to the studies we have carried out for the whole of our profes sional careers, ever since the mid-1970s when human genes became available through cloning. It has always been our ambition to use normal genes in this way, particularly | i^Mdhood diseases sul ) and Friedreich at

Before I came to Australia, I had carried out one of the few gene therapy therapeutic trials for treatment of cystic fibrosis, using lipids as carrier molecules. The trial was a scientific success, although gene expression lasted for only a limited time. It was the beginning of a voyage of discovery, and will still take several years before it is a fully effective therapy. When I succeeded David Danks as the Director of the Murdoch Institute in 1995, it was logical to use the excellent clinical and scientific resources of the Institute to introduce a new team to study gene therapy. Gene therapy is one of the most exciting areas in bio medical research today. When it was first discussed, it was assumed that gene therapy would be directed to cancer and severe disorders that result from single gene defects. While these remain important targets (over two-thirds of the approximately 500 or so gene therapy trials worldwide are for cancer, with rpany of the rest for CF or metabolic diseases), it is also clear that treatment for adult onset multifactorial diseases such as high cholesterol, or prevention of Alzheimer disease, rnay be possible,

Even though the decision to start gene therapy research at the Murdoch Institute was taken soon after I arrived, and endorsed by the NH&MRC review in 1996, it has taken time to decide where best to tar get our efforts. Should we continue research on cys tic fibrosis? We have a lot of experience in this field, and patient care is excellent, but given the intense international competition we decided to help CF research and care by screening for carriers in Victoria, and by helping Brandon Wainwright in Brisbane in his trials making accurate animal models for CF gene therapy. We decided that our gene ther apy efforts here in Melbourne would target two dis eases, Friedreich ataxia and thalassaemia. We have been fortunate in recruiting Panos loannou, since he has great experience studying the molecular genetics of thalassaemia and ataxia, as well as being a pio neer in generating libraries of large human genomic fragments in bacterial vectors (BACs and PACs). I am delighted that Panos has agreed to stay on at the Murdoch as Group Leader of the Gene Therapy Group and make his expertise available for our gene therapy effort, as well as for making these key BAC/PAC library resources available to Australian researchers.

Friedreich ataxia (FA) affects approximately 50 peo ple in Victoria. Since the gene was isolated about two years ago, Martin Delatycki and Sue Forrest have studied every one of these individuals as well as many sufferers from other States. The disease is recessive, which means that carrying one copy of the mutant gene presents no problems; FA develops only when a person inherits a mutation from each parent. When this happens nerves begin to die, leading to unsteadi ness, difficulties with fine muscle control and speech, and eventually heart muscle failure. Therefore, FA is a good candidate for gene therapy - it is possible that a copy of the normal gene may correct the problems - but HOW can we get genes to the brain and ner vous system? Tracy Evans-Whipp, Louise Wangerek and Kumaran Narayanan are transferring genes to nerve cells in culture, each using different approaches. Tracy is using vectors based on Herpes simplex virus (HSV) to deliver the FA gene to patient neuronal cells. HSV naturally infects neurones where it establishes a life long latent infection without disrupting the normal cell function. The approach adopted is to use HSV amplicon vectors in which DNA plasmids containing the minimal viral sequence elements necessary for repli cation and packaging are incorporated into viral capsids. These vectors retain the ability of the virus to readily infect neuronal cells but do not contain any of the potentially harmful viral genes.

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By using genomic constructs spliced to reporter genes such os firefly luciferose and bacterial lacZ, Kathy Williamson and Sherry Cook have been trying to find out which genomic sequences control the expression of the FA gene.

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Louise has used liposomes, the approach I used for CF in London, while Kumaran has tried both mRNA as well as large genomic constructs such as yeast and bacterial artificial chromosomes. Liposomes have been used as a means of delivering nucleic acids into cells in culture. Cationic liposomes associate with nucleic acid through an ionic interaction, allowing the DNA or RNA to cross the cell membrane into the cell. Commercially available liposome formulations designed for the delivery of DNA or RNA into cells have been used on both immortalised cell lines and primary cell cultures. The conditions for delivery of DNA have been optimised for several cell lines as well as mouse primary cortical cells. Optimal deliv ery was determined by altering the amount of DNA added to the cells and varying the ratio of DNA to liposome. Four related but distinct liposomes were used during the optimisation. The optimal conditions determined can then be used in experiments to further increase the efficiency of delivery.

In addition to his clinical duties, Martin Delatycki is looking at delivering genes using the harmless por tion of tetanus toxin, a feasible theory which we are finding difficult to put into practice. He has also stud ied a most interesting finding - the gene that is mutat ed in Friedreich ataxia may cause accumulation of iron in mitochondria, poisoning them in a way which reminds Murdoch staff of problems with other heavy metal disorders such as Menkes disease. Martin is therefore testing the amount of iron in fibroblasts (cells grown from skin biopsies) from peo ple with FA compared to unaffected controls. In addition he is looking at the amount of iron in mito chondria isolated from these fibroblasts, and whether these cells are more susceptible to damage from iron overload. This is of importance not only in looking at the underlying mechanism of the disease process, but also in thinking about approaches to treatment. For example, treatment aimed at leaching iron from mito chondria (iron chelation) may be possible, if it can be shown that iron accumulation is a major problem in humans with FA. It is not enough to get a normal copy of a human gene in to a cell - it has to be controlled biologically in an p propriate way.' We b“elTeve that Tra nsfe rri ng the erftire gene, with all its native surrounding pro moter) sequences and introns, may well function bet ter thdn transferring a cDNA clone driven by a foreign promoter. I

As the tissues that are most affected in Friedreich ataxia are the nervous system and heart, it is not sur prising that clinical trials will be difficultl The brain is isolated from the rest of the body by the blood brain barrier, and few experiments can be carried out on the heart of a living person, particularly one who is already unwell. Until recently, it was extremely diffi cult to find animal models for human disease, until transgenic techniques for mice were developed. We are currently generating a mouse with Friedreich ataxia with which we can investigate the efficacy of FA gene therapy; Kate Elliott and Lachlan McDonald are attempting to clone a naturally-occurring human mutation into the mouse FA gene. Panos loannou has always believed that one strategic approach to gene therapy is direct correction of the mutant DNA sequence using endogenous repair enzymes, but attempts to reproduce data from anoth er group proved frustrating and impossible, here as in many other labs. In light of this, he has resumed approaches using BACs directly for gene expression. Dianne Beck has been working with Panos to make the BAC library a truly useful resource, not only to the Murdoch, but Australia-wide. Stuart Beattie has begun studies on the b-globin gene, which does not function correctly in one major form of thalassaemia. In the long I lent the work q rter _________ __ ...ilia f^-and wifh^ genes may Bad to possil^ itments whieft'^l use human get ome material exclusively, ratherlthan expressing ( onstwcts d on viral promoterl and other elements l4ik«l | rman cDNAs. I ; A’’

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Gene therapy efforts up to now have concentrated on the construction of synthetic vectors carrying the essential coding sequences and little if any of the endogenous regulatory sequences, thus failing to achieve tissue specific regulated expression of the gene of interest. While this approach may work for some types of cancer where cell killing is the aim, through overexpression of the delivered gene, it is unlikely to succeed for most genetic diseases. The development of the BAG and PAC cloning sys tems in the last few years and the construction of high quality genomic libraries is having a major impact on gene mapping and identification as well as human genome sequencing. The large average insert size of the clones, their stability and the ease of handling and purification make BACs more convenient than YACs for many purposes. The rapidly increasing availability of intact functional genetic units and large regulatory regions in fully characterised and sequenced PACs and BACs may be expected to have a major impact on approaches to gene therapy by gene replacement over the next few years. The tools and the techniques are becom ing available for delivering intact functional genes from such clones into cells as episomes, while tech niques for the creation and efficient delivery of human artificial chromosomes or for targeted inte gration could transform the whole area of gene therapy.

Yca» <3ENES 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

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Kumaran Narayanan and Katie Allen are working on gene therapy for the copper disorders Menkes dis ease and Wilson disease. These diseases have been studied at the Murdoch for many years and the research spans the gene therapy team headed by Panos and myself, and Julian Mercer's group on cop per metabolism. Andy Choo is also exchanging tech niques with Panos and myself, since we share a com mon objective, using human genomic constructs for gene therapy. If we have a chance to expand our interests during the coming year, one area to which we share a com mitment with our colleagues in the Royal Children's Hospital is gene therapy for leukaemia. Cancers are a major cause of paediatric death. While therapies for childhood cancers have improved over the past twenty years, especially for leukaemias, there are still many children who do not survive, or who have to go through extremely difficult and unpleasant courses of treatment. I have always thought that leukaemias such as acute lymphoblastic leukemia would be an excellent target for gene therapy, as there is a long tradition of incremental therapies for cancer, and gene therapy would be seen as adding to existing methods of care.

OF FAMILY LIFE. INSURANCE COMPANIES ARE BEGINNING TO PROPOSE THAT RESULTS OF GENETIC TESTS SHOULD BE MADE AVAILABLE TO THEM.

The one thing on which most scientists and doctors agree is that there should be no attempts to carry out gene therapy which could potentially alter inheri tance. We don't want to engineer egg or sperm cells - it could have unexpected consequences, and would move too easily into eugenics rather than treatment. Indeed, we are acutely aware that safety has to be considered above all else when talking about gene therapy, and 1 am pleased that Julian Savulescu, who will be joining the Institute shortly as our Professor of Ethics of the New Genetics, will be studying the ethics of gene therapy in addition to other key areas in this critical field.

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.

■rp Bob Williamson Panos loannou Kate Elliot Tracy Evans-Whipp Kathy Williamson Katie Allen Martin Delatycki Kumaran Narayanan Louise Wangerek Stuart Beattie Dianne Be Sherry^Gook Lachlan McDondlST

Group Leader Group Leader Postdoctoral Fellow Postdoctoral Fellow Postdoctoral Fellow Clinical PhD Scholar Clinical PhD Scholar PhD Scholar PhD Scholar Research Assistant Research Assistant Research /^sistant Research Asslstoftf J:;

FEAR 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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The gene discovery group was formed in 1996 to use current genome techniques to map both rare childhood genetic disorders and the gene mutations which interact with the environment to cause more common dis eases of later life. This offered the opportu nity to use the excellent, but underutilised, clinical resources held by the VCGS to iden tify mutations causing a number of single gene disorders occurring in Victorian fami lies. In addition, in 1997, a successful application was made to the Federal Government to fund a Collaborative Research Centre (CRC) for the Discovery of Genes for Common Human Diseases. It aims to identify and commercialise genes that are important in determining susceptibil ity to common human diseases which are under-served by current treatments. There are five major research programs in the CRC - the key program area for the Murdoch Institute is autoimmune disorders with a par ticular emphasis on atopy.

The gene discovery group therefore has two major aims: ■ to study neurological/neurodegenerative disorders with the aim of determining the underlying genetic defects, investigating possible new methods for treatment and helping to develop these

Gene discovery began in an embryonic way in 1996 with an enthusiastic clinician, Karen Dunn, starting her laboratory career working on defining the gene(s) involved in early-onset childhood eczema. Eczema affects many children, perhaps up to 5% of the pop ulation, is itchy and distressing, and is difficult to treat effectively. Families with two affected sibs and a well defined clinical phenotype were collected in collabo ration with David Hill, head of the Allergy Centre at the Royal Children's Hospital. Markers were initially tested by Karen Dunn and Libby Fitzpatrick in defined candidate regions that had been identified for famil ial asthma, especially the loci at 5q31 (including a major cluster of interleukin genes) and llql3 (the high affinity FCe receptor). Linkage was identified with the chromosome 5 region but not with chromo some 11. Refinement of the region of interest on chromosome 5 will allow identification of candidate genes for mutation sequencing and further analysis. A full genome scan will also be undertaken to deter mine additional loci contributing to the phenotype of childhood eczema, in the hope of identifying those at high risk. These parents could use a mixture of aller gen avoidance and early treatment to prevent severe eczema starting, in line with our general commitment to prevention of diseases of children through our genetic knowledge.

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Trinucleotide repeat identification We have a particular interest in disorders that may be caused by trinucleotide repeat expansion with a focus on traditional single gene disorders. Many disorders with some level of neurodegeneration, such as Huntington disease, show the phenomenon of antici pation where the age of onset is reduced and the severity of the disorder increased in successive gener ations. This is due to instability of trinucleotide repeat sequences. Ivan Biros is optimising methodology to identify and clone trinucleotide repeats such that we can apply the technology to a number of candidate disorders. One disorder we wish to study in detail is Meniere's disease, which is characterised by vertigo, tinnitus and deafness. In a study of a series of patients in England studied by Mr Andrew Morrison in collabo ration with Bob Williamson, anticipation was clearly evident. Therefore we believe this is an excellent can didate for applying trinucleotide repeat detection methodology.

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Two new PhD students joined the group this year. Jan Fullerton is taking a novel approach to define the gene for the eye disease keratoconus by collecting samples from a founder population in Burnie in Tasmania. In collaboration with Simon Foote's labo ratory at WEffI, Jan has completed a full genome scan using 360 markers. She has found five regions where all affected individuals share an allele at a genetic marker. A tempting candidate gene is present in one of these regions and Jan is currently refining her mapping data in addition to studying the candi date genes in more detail. Tom van Agtmael, a PhD student from Belgium, has the challenge of defining the genetic basis of lefthandedness. Mijch of Tom's time initially hcf been spent evaluating;' genetic models for left-handedness from the many described in the literature and defining the best family structure to work with. In addition, a number of genes are now being identified that alter the position of particular body organs in situliin the body when mutafed. These are interesting candidates for involvement ih the genetics of left-handedness and Tom will study each one systematically to inc&de or exclude their involvement,

Trimethylaminuria A long-standing collaboration with Dr Eileen Treacy (Montreal) and Dr John Cashman (Seattle) finally came to fruition in 1997 with the identification of mutations in the candidate gene, flavin mono-oxyge nase 3 (FM03), as the cause of trimethylaminuria, or fish odour syndrome. A group of Australian patients first identified by Professor David Danks were studied initially by Eileen Treacy when she worked at the Institute three years ago and this collaboration con tinued on her return to Montreal. We defined the exon/intron structure of the gene and identified two mutations that are common and have an English ancestry. Expression studies were performed to con firm that the mutations were indeed responsible for causing the disease. Not only are mutations in this gene causative of TMA, but it appears that polymor phisms may result in altered ability to metabolise many important drugs. What started as a study of a very rare and "offensive" disorder may have defined an important new drug polymorphism which accounts for adverse reactions to some pharmaceuticals.

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With the identification of the FRDA gene in 1996, Martin Delatycki and Damien Paris began to charac terise the mutations in all FA patients in Victoria. Most cases of the disease are due to a dramatic expansion of the triplet GAA in intron 1 of the FRDA gene, from a few copies of the repeat to several hundred. We were able to show that the shorter the repeat expan sion (of the smaller allele), the more mild the disease. In general, the repeat size decreases when the muta tion is transmitted from father to child, which is unusu al for trinucleotide repeat disorders. The one excep tion was a male carrier with an unusually small expansion, which was larger in sperm and then larg er still in the affected offspring. This gives clues as to when the triplet expansion occurs. In addition, affect ed individuals in four different families have been found to carry only one expanded allele, the second allele containing a point mutation in the FRDA gene rather than an expansion. Three of these mutations in the Australian population are completely novel, and will help our understanding of frataxin function.

Sue Forrest Karen Dunn Elizabeth Fitzpatrick Melanie Knight Damien Paris

Jan Fullerton Tom Van Agtmael Fanne Galjaard

Group Leader Clinical Fellow (to October 1997) Research Assistant Research Assistant (from October 1997) Research Assistant (to April 1 997) PhD Student PhD Student Visiting Medical Student

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Severe disorders of the mitochondrial respi ratory chain result in neurological or multi organ disease in approximately 1 in 5000 children. These can include developmental delay, movement disorders, liver and heart failure and muscle weakness. There is also increasing evidence that mtDNA mutations or respiratory chain enzyme defects con tribute to common adult-onset diseases such as diabetes, heart disease, movement disor ders and degenerative conditions such as Parkinson and Alzheimer disease.

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These disorders of energy generation can be caused by nuclear or mitochondrial (mtDNA) gene defects and the inheritance can be autosomal dominant, autosomal recessive, X-linked or maternal. Given the complexity of over 100 candidate nuclear genes, it is not surprising that to date only one family in the world has had a nuclear respiratory chain gene mutation identified. Our main research efforts are directed at identifying the nuclear gene defects that cause respiratory chain dysfunction, clarify ing the genetics of mtDNA inheritance, and determining the role of mitochondrial dys function in "common" diseases.

Much of our work focuses on cell lines from patients with respiratory chain enzyme defects. Patients are first diagnosed by the VCGS Metabolic Laboratory. The exact defect is then identified by studying the amount of each enzyme and whether it functions nor mally. Further studies involve somatic cell genetics, in conjunction with Kerry Fowler from the Mouse Model Unit and Marjorie Crawford from the Tissue Culture Laboratory. We have constructed "cybrids" (cytoplasmic hybrids) from patient cell lines with respiratory chain complex IV (cytochrome c oxidase) deficiency. These are cells that contain mtDNA from the patient but a nuclear genome from a healthy person. Results of cybrid studies and mutation analysis (done in conjunction with Henrik Dahl's group) of the 10 nuclear gene sub units of complex IV indicate that the patients have a nuclear-encoded defect but apparently not in the sub unit genes. We have therefore focused on a sub group of patients from 10 Lebanese families with complex IV deficiency, in which the parents are relat ed (i.e. consanguineous). When two Lebanese patient cell lines are fused together, the fused cell (or heterokaryon) is still defi cient, but fusion of a Lebanese patient cell line with one from an Anglo-Australian patient corrects the defect. Such studies are known as complementation analysis, and they imply that most of the Lebanese families have mutations in the same gene.

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Henrik Dahl's group is attempting to identify this gene by homozygosity mapping of these families (in collab oration with Dr Simon Foote at the Walter & Eliza Hall Institute) to identify a genomic locus for complex IV deficiency. Several possible loci with increased homozygosity have been identified and are being studied in more detail to confirm if one contains the causative gene in these families. Similar studies are being done on cell lines from patients with complex I deficiency, which is the most common type of respiratory chain defect in children. Of 47 families with complex I defects, we have iden tified mtDNA mutations in seven families to date. The lower incidence of consanguinity in these families (—10%) compared with complex IV deficiency (—40%) suggests autosomal recessive inheritance is less common in complex I deficiency. A slight male excess among our group of complex I patients sug gests that some patients may have an X-linked defect. A complex I subunit gene on the X chromosome was identified last year, but collaborative studies with Dr Manfred Wehnert (Greifswald, Germany) have failed to identify any patients with mutations in this gene.

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The HIV-1 viral protein Vpr is involved in viral patho genesis by influencing nuclear-localisation of the virus as well as cell division and survival. Vpr expression in mammalian or yeast cells blocks cell division. In con junction with Dr Ian Macreadie (Biomolecular Research Institute), we have shown that yeast express ing low levels of Vpr exhibit gross mitochondrial dys function. Both the growth arrest (induced at higher Vpr concentrations) and mitochondrial dysfunction require a carboxyl terminal motif of Vpr, and we sug gest that the growth arrest is secondary to mitochon drial dysfunction and apoptosis. Mitochondrial dys function has been reported in lymphocytes and other cells during HIV-1 infection, so further characterisa tion of the pathogenic mechanism may aid the devel opment of new strategies to overcome AIDS. Interestingly, toxicity of the anti-HIV drug AZT is due to inhibition of the mtDNA gamma-polymerase, and excessive doses of AZT result in symptoms of mito chondrial myopathy.

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—HI Mitochondrial dysfunction in "common" diseases The mitochondrial respiratory chain is essential for energy generation in nearly all cell types, so it is not surprising that mitochondrial dysfunction could be a risk factor or primary cause of some forms of degen erative disease including diabetes, heart disease and neurological disorders such as ataxia, dystonia, and Parkinson disease. We have studied respiratory chain function in four collaborative studies of diseases in which mitochondrial dysfunction had not been thought to play a major role.

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Insulin secretion defects are common in patients with mtDNA mutations. It is also known that in other tis sues such as liver, kidney and brain, the mitochondr ial energy generating capacity remains very low until late pregnancy or the postnatal period. Our prelimi nary data shows that pig and rat fetal beta cells have an unusua I profile of respiratory chain enzymes, with one enzyme complex being particularly low. We expect that further enzyme and functional studies will show that inadequate respiratory chain capacity is responsible for the fetal beta cell being unable to secrete insulin appropriately in response to glucose challenge.

Diabetes

The insulin-producing beta cell from the fetal pan creas is unable to secrete insulin appropriately when blood glucose levels increase. This immaturity can result in clinical problems such as transient neonatal hyperglycaemia or diabetes. Dr Bernie Tuch's labo ratory (Prince of Woles Hospital, Sydney) has worked on the glucose|nsulin signalling pathway ill fetal pancreas for some years and excluded most bf the potential causes of this immaturity apart from mito chondrial energy generation

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Movement disorders

We have identified respiratory chain defects in a num ber of patients with ataxia or dystonia. The Gene Therapy Group is studying Friedreich ataxia as a model disease for neurological gene therapy. It now appears that the gene product (frataxin) is involved in mitochondrial iron transport, and so it is important to characterise its function in order to develop the most effective therapeutic gene constructs. We dfe thus assisting in characterising mitochondrial function in Friedreich ataxia patient cell lines.

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■ Anti-cancer drug resistance. Ovarian cancer has a high death rate, largely due to the development of acquired resistance to the main chemotherapy agent, cisplatin. Mitochondrial dys function has been reported to be associated with cis platin resistance and decreased cellular drug uptake. In conjunction with Dr Peter Parsons (Queensland Institute of Medical Research), we showed that a cisplatin-resistant human ovarian cancer cell line had a global respiratory chain defect, particularly affecting complexes IV and II. The cell line was extremely sen sitive to killing by various mitochondrial inhibitors and gold-phosphine compounds. This suggests a potential therapeutic strategy of co-treatment of patients with cisplatin and a low dose of a mitochon drial inhibitor (such as AZT). The rationale is that the mitochondrial inhibitor may act preferentially against the rare mutant cells that develop cisplatin resistance, and block their proliferation.

David Thorburn Denise Kirby

Phillip Marie-Jeanne

Unit Head Research Officer/PhD Student Honours Student

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if Early embryonic development - the first twelve weeks of human gestation-is a time of unparalleled change. These changes can never be repeated, and small errors at this stage magnify as tissues develop. Embryos literally remodel themselves while they grow, a process called morphogenesis. The immensely complex structure of the fullterm fetus must be created from an almost featureless collection of cells. To do this, em bryos use techniques akin to origami and clay modelling.

Organs are remodelled by folding, and sculpted by adding tissues, and even by transferring parts^ groups of cells - from one site to another. But embryos also use the stonemason's technique of removing tissue by cell death to throw into relief new shapes in the remaining tissues. While these elaborate problems of shaping are being carried out, the embryo's individual cells are also changing their types, becoming specialised for their future roles, a process termed cell differentiation. And of course, morphogenesis and differentiation must be coordinated so that the cells which assemble to form an organ shaped, for example, like the spinal cord are cells which can become the nerve cells appropri ate for a spinal cord. The orchestration of early development is so complex that it is not surprising that errors are common . Over 3% of live births bear mal formations due to errors in morphogenesis and dif ferentiation, but many more exist as miscarriages.

The nervous system and the face are laid down in the fourth and fifth week in human embryos, and are the most frequent sites of morphogenetic abnormalities. Two important morphogenetic processes are involved in generating these structures: cell migration, which transfers cells from one region to another, and pro grammed cell death, which kills cells in regions where they are no longer needed. Neural crest cells, which form most of the facial tissue and peripheral nervous system, show both these processes. Neural crest cells arise in the neural tube, the struc ture that later forms the central nervous system. Crest cells originating in the future brain mostly migrate to form facial structures. However, at particular sites in the brain, neural crest cells suicide before migration, and are instructed to do this by BMP-4, a short-range hormone. This cell death is thought to divide the craniofacial region into modules which later form separate parts of the face. In contrast, neural crest cells from the future spinal cord are instructed to detach from their origin and migrate away. This instruction is also supplied by BMP-4.

Our starting hypothesis was that the two fates of neur al crest cells, migration or death, were mutually exclu sive, so that increase in one would be at the expense of the other. We therefore tested the effect of BMP-4 on neural tube/crest precursor cells in our tissue cul ture assay. The results are intriguing because they did not sup port the starting hypothesis and instead they showed that the number of migrating neural crest cells and the number of suiciding neural crest cells were not inversely related; BMP-4 increased both in parallel . This increase was in fact at the expense of a third cell population, that of the neural tube. In addition, at least some of the cells fated to suicide could be res cued by allowing them to stick to connective tissue. From this we propose a novel scheme to account for the dual responses of crest cell migration and crest cell death, and the role of BMP-4. We suggest that BMP-4 specifies neither migration nor death, but stim ulates the detachment of cells from the neural tube. If the cell can replace the lost attachment to the neural tube with an attachment to connective tissue, it will survive and migrate. If no attachment can be made, the cell suicides. These findings represent a substan tial change in the view of how the balance between cell death and cell migration is achieved, a balance which is crucial to the proper development of the facial region and the peripheral nervous system.

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Iiii

Cell migration and differentiation in the formation of nerves in the bowel: roles in Hirschsprung disease We have now shown that GDNF transiently induces migrating neural precursor cells in the intestine to divide, but then stimulates their differentiation into nerve cells which can neither divide nor migrate.

In ffirschsprung disease, nerve cells are absent from the anal end of the colon, thus causing gross accu mulation of waste in the bowel. Neural crest cells migrate into the oral end of the gastro-intestinal tract and then spread as a wave to the anal end during early development (weeks 5-12 in humans). These cells then differentiate into the nerve cells which con trol intestinal function.

ET-3 has a subtle effect: while not a growth factor itself, it potentiates cell division and reduces the nerve cell differentiation induced by GDNF. ET-3 also modifies the action of ofher factors like the differenti ation factor CNTF.

Loss of the GDNF and ET-3 growth factor systems give the ffirschsprung disease phenotype, but how this occurs has not been clear.

The ffirschsprung disease phenotype in experimental animals can be achieved by reducing the number of dividing and migrating cells. We propose that loss of GDNF function causes ffirschsprung disease by directly reducing nerve cell differentiation. Loss of ET3 function paradoxically causes ffirschsprung disease by prematurely increasing the differentiation of the dividing and migrating precursor cells into non-divid ing and non-migrating nerve cells. This increase is in a rapid and unmodulated fashion, leaving foo few cells to complete the population of the anal levels of the intestine.

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Chondroitin sulphate proteoglycans have been known for decades; they comprise a large molecular family and are widely distributed in adult and embry onic tissues. They have long been known to play a role in mechanical properties such as compressibility of adult cartilage. Chondroitin sulphate proteoglycans are expressed at sites of loss of cell-cell adhesion both during verte brate development and carcinoma metastasis. We have tested five different chondroitin sulphate proteo glycans for their effect on cell-cell adhesion using assays we designed for this purpose. We found fhat these molecules rapidly destabilised pre-existing cell cell adhesions, the ability varying with the proteogly can type and the cell type.

This suggests that these large molecules have a new function, that of signalling cells fo de-adhere from their neighbours. The specificity of fhis novel function may explain why there are so many large proteogly cans, because the long-known simple mechanical functions of large proteoglycans do not require such diversity. This study has revealed how a vital, but pre viously almost completely unexplored, function during morphogenesis can be controlled at the molecular level. We believe these findings will have importance in the understanding not only of morphogenesis, but also of how cells detach from carcinomas during the spread of invasive cancers.

Don Newgreen Joseph Minichiello Peter Farlie Richard Kerr Catherine ffearn Lyn Rowley Ami Ben-Yaacov

Group Leader Research Assistant Postdoctoral Scholar PhD Scholar PhD Scholar Student Visiting Research Scientist from 1997

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Transgenesis, the introduction of new genet ic material into mice, is of great significance at the Murdoch Institute. Recent advances in this technology allows the generation of pre cise mouse models of human disease. Besides providing great insight into the dis ease process, these models also allow the development and testing of suitable therapy strategies in animal models. It is this criteria which must be met before these candidate therapies can be trialed in human beings.

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Over the past year the increase in research projects has greatly increased the demand tor transgenic mice, where the genome has been changed to pro vide an accurate model of a human genetic disorder or dysfunction. The mouse breeding facilities at the Royal Children's Hospital have become overcrowded and it has been necessary to house a number of mice off-site. During the year the Murdoch Institute, in conjunction with the Royal Children's Hospital Research Institute (RCHRI), were fortunate to secure additional mouse room space in The Institute for Food and Land Resource, Department of Agriculture at the University of Melbourne. This, and the appoint ment of Rachael O'Dowd and Anick Sylvain to help generate and care for researcher's mice, has enabled the transgenic model laboratory to continue to func tion well whilst planning is undertaken for an exten sion to the mouse breeding facility at the RCH site. During 1 997 the transgenic unit was very involved in generating and studying mice which are unable to produce centromere-binding proteins A, B and C as well as Incenp. So far the CenpB and CenpC mice have been carefully studied with Andy Choo's group to look for chromosomal abnormalities associated with meiosis as well as mitosis. Interestingly, adult mice that lack CehpB p^^^^ are only mildly affected, being! smaller in size than normal mice winereas CenpC null mice are unable to survive the first few days of pregnancy. Similar approaches with CenpA and Itpcenp mouse embryo stem cell lines ar^, being

In some cases it is not necessary to generate a mouse mutant in the laboratory because nature has already done the job in providing over 1000 mouse mutants whose causative genetic mutation is unknown. We have excluded the possibility that some of these mouse strains may have mutations in CenpA or Incenp genes by mapping the mouse CenpA gene to chromosome 5 and Incenp to chromosome 19. This work was warmly received at the Mouse Genome conference in Florida this year. Andy Choo, Julian Mercer, Bob Williamson and Panos loannou have utilised the facility to begin gene therapy studies in mice. While Andy's group are test ing candidate artificial chromosomes in mice, Julian, Bob and Panos are introducing genes into mouse breeding lines that have Menkes disease (brindled mouse mutant) and Wilson disease (toxic milk mouse mutant) with a view to correct the phenotype in these mice. During the year Joanne Hill transferred from the transgenic laboratory to assist Julian's group with these important studies. Work continues with Henrik Dahl's group, in collabo ration with Dr Jeff Mann at the Beckman Institute in California, to characterise "transgenic" mouse mod els for mitochondrial disease. The mice generated thus far are abnormally oversized and are being care fully studied to explore any possible role that mito-

chondria Pay play in obesity. Experiments are also under way with Bob Williamson's group to generate mouse models for neurogenetic disorders such as: Friedreich ataxia to study gene therapy strategies. . . In addition the transgenic unit has provided expertise and education for several groups of university stu dents and researchers within the RCHRI to develop models for human inherited diseases and to draw up a Standard Operating Procedures document to assist researchers and members of the RCH Ethic's Committee in matters relating to mice and research.

External collaborations ■ Kennedy disease mouse model (with Gary Warne's Endocrinology and Diabetes group at Centre for Hormone Research, RCH and with Jeffrey Zajac's group at the Department of Medicine, Royal Melbourne Hospital) ■ Candidate genes associated with Wilm's tumour and leukaemia (with Peter Smith's Clinical Haematology and Oncology group, the RCHRI and Department of Paediatrics; and with Marie Dziadek's Anatomy and Cell Biology group at the University of Melbourne) ■ Genes involved with male sterility (with John Hutson's Surgical Research Laboratory at the RCHRI and Department of Paediatrics)

Kerry Fowler Sophie Gazeas Joanne Hill Rachael O'Dowd Robyn Breslin Anick Sylvain

Kerry Fowler

Senior Research Officer Senior Technical Officer Research Assistant (to April 1 997) Research Assistant (from April 1997) Technical Assistant (part time) Technical Assistant (part time; from December 1997)

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ims The Tissue Culture Laboratory staff also train other staff and students in basic sterile techniques, an essential skill if they are to prepare their own cultures as a component of their project. The congestion problems of 1996 have been partly alleviated since a second sterile laboratory was set up. This has been used constantly, as have the extra incubators used for growing cell cultures.

issue Culture Laboratory

Three small rooms tucked away at one end of the South wing grow cell cultures, essen tial tools for research projects throughout the Institute. Maintaining a steady 37 degrees Celsius, the human body temperature, the "cell room" houses the cultures which have been prepared in the Tissue Culture Laboratory. The laboratory is responsible for establishing cell lines from skin or blood samples.

Once established these cell lines are used EP.

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for diagnosis and research by various groups within the Institute and the VCGS - to look for mutations, chromosome abnormal ities and to analyse enzyme levels which may indicate a metabolic disease.

The number of blood samples increased from 50 in 1996 to 106 in 1997. This increase can be attrib uted in the main to samples for the hereditary colon cancer gene analysis being carried out by the Diagnostic DNA laboratory. The number of skin fibroblast lines received remained similar to 1996 (207 and 232), with about half of these received as cell lines established elsewhere. The huge variety and number of tests for various diseases which need to be carried out means that no one facility can offer them all, which is why we receive established cell lines from other laboratories while we, in turn, send many of our cells lines both interstate and overseas. Many of the cell lines we establish or receive from other laborato ries need to be stored long term at minus 196 degrees Centigrade in liquid nitrogen. We currently have about 16000 ampoules stored from almost 7000 different cell lines dating back to 1972. This allows us to store samples from patients with genetic disorders which cannot be studied at present, to await future research breakthroughs.

Marjorie Crawford Tiffany Symes Alison Blake Karyne Lord Zoe Anderson

Scientific Officer Research Assistant (part-time) Technical Assistant (part-time) Trainee Research Assistant (February - August 1997) Trainee Research Assistant (from September 1997)

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Carole Webley and Jane Halliday collated and analysed the data sent from four cytogenetic labora tories on all amniocentesis (AM) and chorion villus samples (CVS) analysed in Victoria in 1996. The lat est prenatal diagnosis report includes tabulated data on 5,718 tests. It was distributed widely to service providers, and to others where requested.

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The main reason pregnant women have an AM or CVS is because of the risk of fetal chromosome abnormality as they become older. Nearly 80% of pregnant women 40 years and over and 60% of women 37-39 years in Victoria have one of these tests. In addition, there were over 300 tests done because of an abnormal ultrasound finding in the fetus, with the follow-up AM or CVS showing that about 20% of these are associated with a major chromosome abnormality. Amongst all women test ed in 1996, 59 fetuses were detected with Down syn drome (trisomy 21), and another 42 with trisomy 18.

Epidemiology is the study of the distribution and determinants of disease and health in populations. The Epidemiology Unit has grown in 1997 with new staff and new pro jects under way. The projects undertaken by the Unit describe health outcomes relevant to genetics in a number of subgroups of the Victorian population. For instance, we have continued to monitor and report on all pre natal diagnostic testing by amniocentesis or chorion villus sampling (CVS) in Victoria and have undertaken to follow-up the first 900 pregnant women screened by the VCGS maternal serum screening quadruple test. Data have als5,beeri collected on children who were didgnbsed by amniocentesis or CVS during pregnarvpy as having en^nusual chromosome complement, V7e have the benefit of expert advice fronr] our dinicdl col leagues, particularly :,Dr “IJs Sheffiel4 who has a long commitment to epidemiolpgical approaches within the VCGS.

One exciting development was the receipt of a VicHealth grant to study women's attitudes to, and knowledge of, prenatal testing for birth defects. This project began in November 1997 and will continue through 1999 and will answer important questions relating to informed choice and access to these tests. The Epidemiology Unit has designed ques tionnaire-based studies to evaluate specific aspects of genetic service provision, particu larly in the field of familial cancer. These studies aim to provide information that will help with planning and provision of health services in the specialist clinic setting, as well as in the broader community.

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The target population are women who are more than 26 weeks pregnant, 37 years and over, and who have not had a prenatal diagnostic test. Answers to the questions posed will help to determine if women are making informed choices with regard to testing or whether there are problems with access in our com munity for women in particular subgroups, such as those of non-English speaking background or those living in rural areas.

Women's knowledge of and attitudes to prenatal diagnosis of birth defects We have recently begun a Vicffealth-funded project on women's knowledge and attitudes to prenatal diagnosis of birth defects. Two new staff members have been employed, Geraldine McDonald as Project Officer, and Rosemary Warren as Research Assistant. So far, ethics approval has been sought and granted from the Royal Women's Hospital and Monash Medical Centre, where a questionnaire is being piloted. We plan to approach eighteen hospi tals altogether throughout the State,

Maternal serum screening The initial phase of the maternal serum screening (MSS) program of the VCGS has been monitored by Susan Nisbet who distributed a short questionnaire to the doctors who referred the first 900 women for screening. Where the doctor could not provide preg nancy outcome information, this was sought from the Perinatal Data Collection Unit in the Department of Human Services. An enormous amount of effort by Susan, staff of the MSS program, and the doctors involved, has resulted in information being gathered on 97% of these tests, so that results will be available in early 1998.

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rms been completed and returned. We will use these data to determine whether people attending the clinic have been given appropriate and understandable informa tion and are satisfied with their attendance.

ii SPINE - spina bifida information network Vicffealth has provided funding so that an informa tion register can be established which contains infor mation on families at high risk of having a child with a neural tube defect. It is now known that 0.5 mg of folate (a B-group vitamin available in tablet form) taken before and during pregnancy can reduce the risk of a neural tube defect in more than two-thirds of cases. This SPINE register will facilitate the dissemina tion of health promotion material, particularly infor mation on periconceptional folate to families who need more folate than the general population because they are in the high-risk category. Inclusion on the register will be through a variety of sources. Robin ffayles, the proiect officer, has been working in the Epidemiology Unit and collaborating with the Spina Bifida Foundation and the Centre for Community Child Health and Ambulatory Paediatrics at the RCH, to determine the best methods to contact individuals and families for permission to be included on the register.

Follow-up of mosaic fetuses A long-term case/control study of children who were diagnosed in utero with a mosaic karyotype has involved design and administration of about 80 postal questionnaires so far. A mosaic karyotype is one where some cells have a normal number of chro mosomes, while others are aneuploid (have either gained or lost a whole chromosome), in a sample from a fetus. The long term health effects of this is usually unknown. Collaboration between Honey Heussler and Elizabeth Waters from the Centre for Community Child Health and Ambulatory Paediatrics was important in the design phase of this project. Honey left for the UK in August and the follow-up is now being continued by our group. This is revealing useful information on the experience of these families and the health of the children. We plan to obtain as many responses as possible and the initial colfeborators will assist with the final analysis.

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Birth defects and use of genetic' | counselling services Veronica Collins is undertaking a part-tirrle PhD which is expanding an area of interest of thd Unit the evaluation of use of genetic services in the com munity. She is studying factors which influence fami lies to seek genetjc counselling services after the birth

of a baby with a congenital malformation. Veronica is comparing families who have, and have not used our statewide genetic service. The preliminary phase of the project has involved interviewing parents of children born in 1994 with Down syndrome or cystic fibrosis. This experience has provided invaluable information which is being used in the design of a structured questionnaire for the main study.

Cancer genetic services We have continued to collaborate in an international study of the psychological impact of predictive genet ic testing for a form of inherited colon cancer, famil ial adenomatous polyposis (FAR). The numbers being recruited into the questionnaire-based study decreased in 1 997 because the backlog of people seeking testing had decreased. We have therefore had time to extend our interest in cancer genetic ser vices by developing and piloting a questionnaire to evaluate aspects of the new Familial Bowel Cancer Clinic (which covers other forms of inherited colon cancer as well as FAR) which has been established at the Roi^r^Meibourne Hospital. 'p

Information derived from these epidemiological stud ies will be published and distributed widely, hopefully resulting in a greater awareness of the health needs of the public, specifically for prenatal diagnosis and the familial cancer genetic service.

Jane Halliday Veronica Collins Carole Webley Honey Heussler Susan Nisbet Geraldine McDonald Rosemary Warren Robin Hayles

We have found that people are very willing dnd able to participate in the study since 89% of 70 pr|-clinic and over 70% of the post-clinic questionnairdi have

Les Sheffield

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Group Leader Research Officer/RhD Scholar Research Assistant Trainee Clinical Fellow (until Aug 1 997) Master of Rublic Health Student Rroject Officer (from Nov 1997) Rroject Assistant (from Nov 1997) Project Officer (jointly with Spina Bifida Foundation) Consultant Medical Geneticist

nn Because of the major advances in information tech nology over this decade, a major project develop ment was initiated in 1996-97. This development was made possible by a joint venture programme undertaken by Telemedia Software Laboratory at CP. This project is researching technology for distribution of multimedia over the internet, applying the Java language in a Windows 95 environment to produce a programme called SILKWORM (System for Illuminating Knowledge of Relevance to Medicine). This programme can be used on the World Wide Web providing limited access to a nominated network of users. An amalgamated POSSUM and OSSUM has been ported into this programme.

POSSUM - Pictures of Standard Syndromes and Undiagnosed Malformations in a com puter database, was developed by the VCGS and the Murdoch Institute, and is now used by clinicians throughout the world to help diagnose children with different patterns of birth defects. It has been ten years since the commence ment of the POSSUM Project, a long and strong collaboration between Dr Agnes Bankier of the VCGS and Computer Power Group (CP), in particular Mr John Marquet. In July 1996 we issued version 4.5, the eighth update. POSSUM sales around the world have continued.

The web residence provides links to other on-line databases (like OMIM) and to the community of Possum users, making net conferencing a possibility for the first time with the option of updating via the internet. The image database of the programme originally on laser video disk, has now been trans ferred to CD-Rom technology have also made it pos sible to transfer Possum's extensive pictorial database to CD-Rom, making it transportable and more easily accessible. For confidentiality reasons, the pictorial database will not be resident on the internet.

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POSSUM Staff Agnes Bankier Anne Cronin Cathy Rose Henny Miller

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The new version of POSSUM 97 will be accessible to a wider community including Macintosh users

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Senior Lecturer in Medical Genetics Sylvia Metcalfe commenced appointed as Senior Lecturer in Medical Genetics at the beginning of 1997, based at the Murdoch Institute but appointed through the Department of Paediatrics of the Faculty of Medicine, in the University of Melbourne. Through Sylvia, the Murdoch Institute has expanded its involve ment in teaching of medical undergraduates. Other members of the Murdoch Institute and the VCGS who are also involved in teaching are Bob Williamson, John Rogers, Jane Halliday and Mac Gardner.

The Education Team was expanded into a formal Unit in February 1997 with the addi tion of Sylvia Metcalfe and MaryAnne Aitken. Key roles included the relief of other staff from the growing number of requests from students and the public for information about inherited diseases and an expansion of our undergraduate educational role in the medical course at Melbourne University.

New Medical Curriculum

Communify Education and Outreach, and Operation Jigsaw

The EDUCATION UNIT HAS FOUR MAJOR FOCUS AREAS:

COMMUNITY EDUCATION OUTREACH PROFESSIONAL EDUCATION PRIMARY AND SECONDARY EDUCATION TERTIARY EDUCATION

MaryAnne Aitken, a fresh graduate of the Graduate Diploma of Genetic Counselling, began with the title of Outreach Officer, Community Genetics. MaryAnne's task is to take genetics education out to the community and promote the work of the Murdoch Institute. Getting Operation Jigsaw underway was number one on the list: a schools education project which included fundraising for the Institute. Promotional material was produced, and a mailoutto all schools was undertaken in liaison with the Ministry for Education. Badges, fliers, posters and a genetics awareness quiz were prepared with generous spon sorship for these expenses from the St George Bank. Operation Jigsaw raised $33,000 for the Institute in its first year and we expect this to grow in 1998, but in many ways the most valuable element has been the increased knowledge and interest in human genetics in many Victorian schools. -

1999 will see a completely restructured medical cur riculum at the University of Melbourne, with major emphasis on an integrated course employing prob lem-based-learning. Sylvia is involved in the devel opment and integration of genetics teaching within the new curriculum with input for all years of the course, but in particular for the first four years. She has been successful in obtaining funding ($43,000 from January 1998) from the University of Melbourne's Teaching and Learning (Multimedia and Educational Technologies) Committee for the devel opment of an interactive multimedia teaching pro gram in medical genetics and will be focussing on this project during 1 998.

Genetics for the People Working together, Sylvia and Mary Anne have also organised workshops in genetics for the lay commu nity and health professionals. The Unit handles the numerous telephone inquiries from students, espe cially for the Genetics component of Year 12 VCE Biology! As Outreach Officer Mary Anne regularly addresses school, professional and community groups. The Unit was fortunate to receive funding for an Administrative Assistant to which Mark Fisher was appointed. Mark's skills with computers are called upon for the preparation of publications, educational material, and maintaining the Murdoch Home Page on the web (http://murdoch.rch.unimelb.edu.au). Mark also handles many telephone inquiries, and coordinates the updating of many of the information booklets on inherited diseases.

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Sylvia Metcalfe, Mark Fisher and MaryAnne Aitken

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Whe Murdoch Web Site HTTP ://MURD0CH .UNIMELB.RCH.EDU.au

A WEB SITE IS OFTEN A POINT OF DISCOVERY FOR WEB BROWSERS. ThE MURDOCH'S OWN WEB SITE IS EXPANDING AND BEING UPDATED ON A REGULAR BASIS BY MARK FiSHER IN THE

Murdoch Institute Commercials Production of three innovative thirty-second television commercials helped to raise community awareness of the Institute and the services which it provides. These were aired as community service announcements and we thank the three network stations which screened them.

Education Unit. The site provides an outline of the Murdoch Institute and the Victorian Clinical Genetics Service. It also offers information on regular seminars held at

CRC Education Officer Medical Research Week Medical Research Week was held in June and an information booth and display was set up in Melbourne Central. This was an overwhelming suc cess, so much so that the programme will be extend ed in 1998.

1998 promises even more growth in the field of Community Genetics with the assistance of CRC input. Ivan Maccoccia, who was a 1997 Graduate Diploma in Genetic Counselling student, has accept ed the position of Education Officer, Cooperative Research Centre for the Discovery of Genes for Common Human Diseases.

MaryAnne Aitken Sylvia Metcalfe Mark Fisher

Education Outreach Officer Senior Lecturer in Medical Genetics Administrative Assistant

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THE Institute; staff vacancies; courses and workshops on offer; educational informa tion ON inherited diseases designed for VCE STUDENTS; LINKS TO SUPPORT GROUPS AND OTHER WORLDWIDE ASSOCIATIONS; AND DETAILS OF THE POSSUM/OSSUM DATABASE. A LIST OF ALL STAFF CAN ALSO BE FOUND ALONG WITH A LIST OF STAFF EMAIL ADDRESSES.

The repertoire fortesting for adult neurodegenerative disorders performed by Ivon Biros and Janet Shaw has been expanded. Addition of each new test means reanalysing a large series of stored specimens where the genetic basis of an individual's disease is not yet known. This is an ongoing process involving families being kept up-to-date with changes in information.

» Diagnostic Laboratories

The requests for carrier testing for cystic fibrosis still represent the major portion of the workload for this disorder (Karina Forshaw) with much interest from families identified early from the newborn screening program.

The DNA Diagnostic Laboratories at the Murdoch Institute and Monash Medical Centre offer genetic testing for prenatal diagnosis, presymptomatic diagnosis and carrier testing for diseases and syndromes such as cystic fibrosis. We provide a com prehensive testing service, using up-to-date methodologies that are constantly under review. Although paediatric disorders were the initial mainstay, the service has expand ed dramatically over the last few years to include many disorders with onset in adult life, particularly neurological disorders such as Huntington disease and spino-cerebellar ataxias, and hereditary colorectal cancer.

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We are well on target to completion of the identifica tion of the family-specific alterations in the adenoma tous polyposis coli (or ARC) gene which causes famil ial polyposis (an inherited bowel cancer) in the 70 families currently known in Victoria, thanks largely to the efforts of Steve Nasioulas assisted by Jenny Douglas and Karina Forshaw. This work has resulted in two publications with a number of other interesting clinical findings awaiting submission. In addition, we collaborated with the Orthopaedic Research Group headed by John Bateman at the Royal Children's ftospital to evaluate a protocol that would improve the sensitivity of detection of mutations that cause premature termination of protein synthesis. During the course of the year, Jenny has turned her attention towards the more challenging task of mutation iden tification in hereditary non-polyposis colorectal can cer where one of at least four different genes could be responsible for the disease.

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One of the major events of the year was preparation for NATA (National Association of Testing Authorities) accreditation. This accreditation ensures that all tests conducted by our laboratories meet national stan dard guidelines. Jan Brasch (Murdoch) and Andrea Twomey (Monash) are to be credited for their excel lent co-ordination of the process. We are pleased to announce that the accreditation was successful with only minor alterations to operational procedures sug gested. One area in which our laboratories needed upgrading was our patient information system, and we have now purchased a data system from Canada to automate this area.

Desiree ■ du Sort

Desiree du Sail joined the Murdoch laboratory in October following the successful completion of her PhD in Andy Choo's laboratory working on the human centromere project. Desiree is also a trained cytogeneticist. She has assumed the post of scientistin-charge since Sue Forrest has moved on to lead the Gene Discovery group, and we are sure the labora tory will go from strength to strength under Desiree's enthusiastic leadership. Iswari Setianingsih has made excellent progress in her PhD towards identification of the spectrum of muta tions causing beta thalassaemia in the Indonesian population. She has learnt a battery of new tech niques which will be beneficial when she returns to her home country, Indonesia, to expand their genetic services for thalassaemia. We have had three sum mer students, Fanne Galjaard, a 5th year medical student from Holland; Lucinda Johnson, a third year science student; and Sarah Foster, a second year medical student. All have been able to learn molec ular techniques quickly and have enjoyed seeing the operations of a fully-functioning laboratory, and con tributing to the research.

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The DNA laboratory at Monash Medical Centre has been heavily involved in thalassaemia gene testing, both because of the increasing heterogeneity and load in this field, and because it fits well with the inter ests of Dr Don Bowden, who is also responsible for the care of thalassaemic patients. Because of deci sions made by the Department of ffuman Services, the role of the Monash laboratory is likely to continue to involve expansion in this area, while other tests may be relocated to the Murdoch Institute site.

On a more personal level, as I "retire" from the Scientist-In-Charge position, I would like to take this opportunity to thank the excellent laboratory staff thaf I have had the pleasure of working with over the past seven years. They are a committed and responsible team and are the essential operational core which enable us to offer such an excellent service. In addi tion, I would like to thank all the clinicians and coun sellors who have made us feel such an integral part of the patient management process.

Sue Forrest

Caroline Bowdich IRS

M Murdoch Institute / VCGS Staff Sue Forrest Ivan Biros Jan Brasch Desiree du Sart Jenny Douglas Karina Forshaw Steven Nasioulas Janet Shaw Tom Milovac Iswari Setianingsih Les Sheffield Fanne Galjaard

Scientist in Charge Scientific Officer Medical Scientist ScientificOfficer (from Oct, 1 997) Medical Scientist Medical Scientist Medical Scientist Research Assistant Technical Assistant PhD Scholar Liaison Clinician Visiting Medical Student

Monash Medical Centre Staff Jean ffendy Andrea Twomey Kathy Garafolo Don Bowden Agnes Bankier

Medical Scientist Medical Scientist Trainee Scientist Clinical Adviser Liaison Clinician

Our bright and cheerful 1997 Genetic Counselling Diploma graduate Caroline Bowdich has been awarded a Queen Elizabeth Trust Grant. This will enable her TO TRAVEL TO LoS ANGELES IN 1998 TO UNDERTAKE TWO MONTHS' WORK EXPERIENCE IN GENETIC COUNSELLING, WITH AN EMPHASIS ON OSTEOGENESIS IMPERFECTA.

Caroline

was born with osteogenesis IMPERFECTA (Ol), A CONDITION WHERE THE BONES ARE FRAGILE DUE TO A LACK OF COLLA GEN IN THE BONE TISSUE, AND NEEDS A WHEEL CHAIR FOR MOBILITY.

Since graduating Caroline has been coor dinating THE "Activism, advocacy and dis ability RIGHTS conference", HELD IN FEBRUARY

1998. Four disability-based groups banded TOGETHER TO ADDRESS EDUCATION, EMPLOY MENT AND THE LAW FOR PERSONS WITH DISABILI TIES. Caroline is also President of Osteogenesis Imperfecta Association.

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Disorders of mitochondrial energy generation are among the most common types of inborn errors of metabolism, with an incidence estimated at 1 in 5000 births. The most severe disorders present at birth with multi-organ failure, and are usually lethal. If appropriate tissue samples are obtained, such patients can usually be diagnosed accurately, because the residual enzyme activity tends to be very low. There is a wide spectrum of severity of these dis orders, however, with milder defects presenting in mid to late childhood, or even during adulthood. Later-onset cases are more difficult to diagnose accu rately as only one or a few organs may be involved, and the residual enzyme activity tends to be higher.

etabolic Laboratory f

Metabolic diseases result from enzyme defects affecting the breakdown and re-utili sation of sugars, fats or protein in the body. Many of these disorders are severely dis abling, but in some cases affected children can lead a normal life if their condition is recognised early. Several hundred metabol ic disorders are known and most are diag nosed either by testing for accumulation of compounds before the defective enzyme or by measuring the suspected enzyme directly.

Children suspected of having an amino-acid metabolism disorder are diagnosed and monitored by the VCGS Metabolic Laboratory. Plasma, urine and cerebrospinal fluid are analysed by a method called quan titative amino acid analysis. This method is also used to monitor over one hundred such children from around Victoria to ensure that diet and drug therapy are maintaining their amino acids at an appropriate level. Our other major diagnostic test is the mea surement of enzymes in patients suspected of disorders of energy generation, for which we act as the Australasian referral centre.

Mitochondrial enzyme activities vary quite widely in healthy people, particularly in muscle, where they are very high in athletes, and quite low in any form of muscle disuse. It can thus be difficult to distinguish real from secondary enzyme defects, and it becomes confusing when centres around the world have differ ent (somewhat arbitrary) criteria for what they regard as diagnostic results. Recently Ed Byrne at St Vincent's Hospital proposed the first objective model for diagnosing adult patients as definite, probable or possible based on assessing the results of clinical, enzyme, histological, DNA and metabolic investiga tions. Francois Bernier, a visiting Canadian Clinical Fellow, spent six months in our laboratory this year reviewing the charts and results of all the 139 Victorian children we had studied in the last 5 years. Based on this review, we have suggested a modified diagnostic scheme for adults and children, which we hope may provide the starting point for consensus international diagnostic criteria. Analysis of metabolites and enzymes is current!/split between the Royal Children's Hospital Biocherpistry Department and the VCGS Metabolic laboratory. Two factors:,;have prompted us to review the current arrangemenf so that over the next year we can devel op a single joint structure that will be d more eff|cient service provider. The first of these is the likely) pur chase of a tandem mass-spectrometer^: which vfould

be used by both the VCGS and RCH laboratories as well as the VCGS Newborn Screening laboratory. The recent appointment of a new Clinical Director of the VCGS and a new Professor of Pathology at the RCH Biochemistry Department has also prompted us to re-assess the current division of services which has evolved over time. Inborn errors of metabolism diagnosed this year include 15 definite and two probable respiratory chain defects, eight (homozygous) cystinuria, four phenylketonuria, five hyperphenylalaninaemia, three pyruvate dehydrogenase defects plus isolated cases of several other inborn errors, including citrullinaemia and the second Australian patient diagnosed with fumarase deficiency.

David Thorburn Ivan Francis Denise Kirby Erin Oldaker Avihu Boneh Francois Bernier

Scientist-in-charge, Enzymology Scientist-in-charge, Metabolism Research Officer Research Assistant Clinical Liaison Visiting Clinical Fellow (to June 1997)

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We have considerable experience in the use of this technique for monitoring the progress of bone mar row transplants and have now introduced its use for testing two neuropathies, namely Charcot Marie Tooth Type lA and its mutational counterpart ffNPP (ffereditary Neuropathy with Pressure Palsies) and for overnight detection of chromosome aneuploidies in prenatal diagnosis, where there is a high fetal risk.

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Lucille Voullaire and Howard Slater

The Cytogenetics Laboratory provides a comprehensive service in routine chromo some testing to hospital and private clini cians throughout Victoria and Tasmania. All types of routine testing for constitutional and acquired chromosome abnormalities are available using banding, fluorescent in-situ hybridisation (FISH) and DNA analysis. Specialised testing using molecular cytoge netic analysis is provided throughout Australasia for non-routine, complex or urgent cases. The Cytogenetics Service works closely with the VCGS and Murdoch Institute, and many research issues that arise can be dealt with quiqkly because of this seamless interface. Advances in scientific testing for various inherited diseases and increased awareness of genetic disease, both by the medical pro fession and the public has led to an increased demand for services of the Victorian Clinical Genetics Sen/ice (VCGS) Cytogenetics Laboratory.

Despite proving to be a very tightly-held market, our prenatal chromosome testing work has increased in a carefully controlled manner consistent with the requirement to guarantee fast turnaround times, train additional staff and investigate new approaches. Many of our urgent cases derive from the VCGS Maternal Serum Screening Programme and ultra sound scanning of nuchal translucency. In the former referral group, gestational age is almost always advanced and in both, the patient is anxious to receive a rapid test result. All revenue received from this testing is invested in new service development.

Our workload has increased steadily over the past few years by approximately 1 0% per annum, but this year we were hit by an increase of almost a quarter and some 5,800 tests were performed. We were fortu nate to acquire the services of two trained cytogeneticists from overseas who have helped us cope with the extra workload.

The Future The gradual move towards using molecular tech’ niques over the past few years would leave few to doubt that the future of clinical cytogenetics lies in this direction. However, it is only within the last twelve months that we have gained a clearer view of what lies ahead. Molecular cytogenetics promises much 1 higher resolution analysis, flexibility to match situa- ' tions with a specific risk with a more appropriate ana- -*>* lytical approach, faster turnaround times and lower costs. The VCGS laboratory is in a strong position to ensure rapid but timely application of these new developments. Cytogeneticist in Charge Howard Slater

Management Group

Vida Petrovic Lucille Voullaire Anne Robertson Lorna Webber

Scientists

Selena Bourke Trent Burgess Melissa Curtis Sue Dale (to July 1997) Julie Davies (to May 1997) Tracy Fleming David Francis Lee Harrison (from November 1997) Louise Hills Sarah Nouri Ralph Oertel Vladimir Pupko (from August 1 997) Marie Thorpe Cathryn Vaux

Technicians

Ian Brooks; Simon Benedetti; Wali Drummond Bo Jezierski;

Research and Development Work The laboratory has focused its research efforts on two ambitious projects. One is the detection of fetal cells in maternal blood as a source of diagnostic material to replace invasive testing by amniocentesis or chori onic villus sampling; the other is the application of Comparative Genomic Hybridisation (CGH) to preimplantation diagnosis in vitro fertilisation (IVF). Both projects use techniques which are within our repertoire but are challenging to apply because they require single cell manipulations.

Changes and Advances 1997 has been the year of Interphase FISH testing, what I call 'cytogenetics without chromosomes'l This new technique allows us to detect specific abnormal ities without having to prepare cells showing visible mitotic chromosomes. This has several advantages in special situations where dividing cells are very rare, a result is needed quickly or other techniques have inadequate sensitivity.

Two papers were published in collaboration with Professor Jenny Graves' group at Latrobe University where the chromosome microdissection facility devel oped by Lucille Voullaire was applied in studies of sex chromosome evolution in marsupials.

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Lynda Phillips.

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Graduate Diploma in Genetic rcounselling The Victorian Graduate Diploma in Genetic Counselling was a VCGS initiative with Melbourne University, starting in 1996. Twenty students have now completed the course, and we have many more applicants for the course than places. The majority of applicants have scientific qualifications, many with higher academic degrees, but others have come from the social sciences, nursing and teaching. We welcome eleven new students, six of whom are studying parttime, to the course in 1 998.

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We interview most of those who apply for the course basing selection not only on acade mic skills, but also on community interest and experience. Apart from this the students are from diverse academic and professional backgrounds, creating a complimentary mix of skills and enabling a strong student group focus to develop. The skills learned in the core subjects of Human Genetics and Counselling are reinforced by a four week supervised practicum. This is arranged in a clinical genetics setting, usually within the Victorian Clinical Genetics Service, but stu dents have also been offered placements at the Anti-Cancer Council of Victoria, genetic units interstate, and the Department of Human Genetics of the University of Witwdtersrdnd, Sotifh Africa. These oppor tunities not only provide valuable training for our students, but also spread the word about our Course to future applicants

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We are delighted that all of those who took the course in 1997 passed well, and the feedback from students was very positive. In response to tutor and student suggestions, we have changed aspects of the course content and assessment techniques in ffuman Genetics in order that students and tutors can moni tor progress in a more objective way. We know that there will be a great need for genetic counsellors during the coming decade, as molecular genetics impacts on adult medical specialities. This need has, however, not yet been translated into posts, and there is still concern at the employment prospects for those we train. New openings are slow ly being created in line with more demand for genet ic counselling as a result of greater awareness of the role of genetics in predisposing to diseases such as breast and colorectal cancers, cardiovascular dis ease and neurological disorders such as epilepsy and Alzheimer disease. The VCGS believes that coun selling is a essential extension of a family's genetic awareness, and we are pleased that the Victorian Discussion Paper on Genetic Services gave strong support to this view.

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Five of our students are now employed in new genet ic counselling positions, but it is equally relevant that several students have returned to their past employ ment with increased relevant counselling skills, and there is potential for more placements as new posi tions are created around the State

Margaret Sahhar Leslie Sheffield I I ■

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Course Convenor Human Genetics Coordinator

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aternal Serum

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Quadruple Test Demand for the Victorian Quadruple Maternal Serum Screening test has escalated in the past year. The quadruple test can be used to give risks for Down syndrome, neur al tube defects, trisomy 18 and some other birth defects. A sample of mother's blood taken between 15 and 20 weeks of preg nancy can help identify fetuses with an increased risk of certain birth defects. Four chemicals from the mother's blood are analysed to determine a "risk figure" for the pregnancy. Because it is a screening test it is not perfect - we expect to detect seven out of ten cases of Down syndrome and four out five neural tube defects. If an increased risk is detected, further diagnostic tests are avail

able.

The Newborn Screening Laboratory (NSL) tests every baby born in Victoria for three rare but important disorders 1. phenylketonuria (PKU)

In our first 18 months of operation we have tested just over 5,000 pregnant women. We were extremely pleased to receive funding in September from the Department of Human Services to offer public patients the same opportunity to undertake this form of screening as those coming privately. We recognise that this type of screening can be stress ful for women whose pregnancies are identified as being at an increased risk of a birth defect, and much of the work of the team is to provide support, infor mation and a counselling service to many women who telephone or visit worried or with questions about the meaning of the test.

Robin Forbes Ivan ■ Len B|nacquisto Mana|lbrahinr)' 'VTiT Les Sheffield ' ' / W' Anne fronin "

CH is a predominantly 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 (cre tinism). In Victoria, CH occurs in 1/3100 births (20 per year). 3. cystic fibrosis (CF) 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 avail able before debilitating malnourishment and respira tory disease has taken hold. The Victorian Newborn Screening Program is one of the integrated services provided by the VCGS. Its emphasis is on early diagnosis and prevention, and takes its part one of the many health services which maximise the opportunities for infants affected with one of these disorders.

Co-ordinator

'S’SftfdrSCfentist Medical Scientist \ Medical Scientist Medical Adviser : Program Convenor;

Len Bonacquisto and Mono! Ibrahim

PKU is an inherited disorder of amino acid metabo lism and occurs in 1/13000 births. When both par ents 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.

2. congenital hypothyroidism (CH)

Ivan Francis Nick Tzanakos Len Bonacquisto Nella Napolitano Mona El-Masri

Scientist-in-charge Medical Scientist Medical Scientist Medical Scientist Trainee Medical Scientist

Huntingtons disease Huntington's Disease (formerly known as Huntington's chorea) is a progressive inherited neurological disorder THAT NORMALLY MANIFESTS BETWEEN THE AGES OF 35-50, ALTHOUGH IT CAN OCCUR (RARELY) IN CHILDHOOD OR AS LATE AS THE EIGHTH DECADE. It CAN AFFECT EITHER SEX AND SYMPTOMS MAY INCLUDE INVOLUNTARY JERKY MUSCLE MOVEMENTS, EMOTION AL DISTURBANCES AND COGNITIVE DIFFICULTIES. ThE DURATION OF

SCSCIAL WORK AT THE VCGS

THE ILLNESS CAN VARY FROM 10-25 YEARS.

Support and advice from social workers has been available

Adult predictive, and prenatal, testing for Huntington's Disease has been offered by the Victorian Clinical Genetics Service since 1989. Initially our predictive tests were by

FROM THE VCGS FOR OVER 20 YEARS. THIS SERVICE IS MADE AVAILABLE TO THOSE FAMILIES WHO SEEK GENETIC COUNSELLING, OR HAVE A CHILD WITH A GENETIC METABOLIC DISORDER, AS WELL AS COMMUNITY CONSULTATION WITH PROFESSIONALS.

LINKAGE ANALYSIS, BUT SINCE 1 993 ACCURATE DIRECT TESTING HAS BEEN AVAILABLE. To DATE OVER 340 CLIENTS HAVE AVAILED THEM

When a genetic disease has been diagnosed families need

SELVES FOR TESTING.

INFORMATION, PRACTICAL AND EMOTIONAL SUPPORT AND ADVICE ABOUT THEIR CHANGED CIRCUMSTANCES AND LIFE PLANS. FAMILY AND INDIVIDUAL COUNSELLING, REFERRAL TO COMMUNITY

The Huntington's Disease Predictive Testing Programme was THE FIRST PROGRAMME THAT OFFERED "WELL" ADULTS THE KNOWL EDGE OF WHETHER OR NOT THEY WOULD DEVELOP HuNTINGTON'S Disease in future years. Because of the possible emotional

RESOURCES, SETTING UP SUPPORT GROUPS, COMMUNICATION, POST-GRADUATE AND STAFF EDUCATION, AND SUPERVISION OF GENETIC COUNSELLORS SEEKING HGSA ACCREDITATION ARE PART OF THE SOCIAL WORK ROLE.

RAMIFICATIONS OF SUCH PERSONAL KNOWLEDGE, THE AIM OF THE PRE- AND POST-TEST COUNSELLING PROGRAMME IS TO BE FLEXIBLE TO THE INDIVIDUAL NEEDS OF THE CLIENTS AND TO PROVIDE SUP PORT AND INFORMATION. THEN, WHATEVER THE OUTCOME OF THE

Currently there are three social workers at the VCGS Margaret Sahhar (counselling, support groups. Convenor Graduate Diploma in Genetic Counselling, STAFF supervision), Sue Mansie (Co-ordinator Huntington Disease Program, post-graduate teaching, staff supervi sion), AND Maureen Crawford (Metabolic Unit, family

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TEST, THE PEOPLE MAY RETURN TO THEIR HOME AND COMMUNITY WITH A RENEWED SENSE OF LIVING THEIR LIVES TO THE FULLEST. THIS COUNSELLING AT THE VICTORIAN CLINICAL GENETICS SERVICE IS MANAGED BY SUE MaNSIE WHO IS A SOCIAL WORKER, FAMILY THERAPIST AND REGISTERED NURSE.

COUNSELLING, STAFF SUPERVISION, SUPPORT GROUPS WITHIN THE

VCGS).

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Although the programme involves a team approach as rec ommended BY THE International Huntington's Disease Association and the World Federation of Neurology, Sue Mansie has adopted an "intimate" approach which emphasis es A CLOSE COUNSELLING RELATIONSHIP BETWEEN THE COUNSELLOR AND THE CONSULTAND. A RECENT STUDY HAS SHOWN THAT THE

CONSULTANDS FIND THIS MODEL OF COUNSELLING TO BE VERY SUCCESSFUL.

Generous support from Dame Elisabeth Murdoch enabled PRODUCTION OF A VIDEO IN 1 997 THROUGH THE AUSTRALIAN Huntington's Disease Association. Entitled "Freedom of Choice A Human Response to Genetic Testing for

Huntington's Disease", the video is available for borrow ing THROUGH THE ■T-

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INSTITUTE, AND HAS BEEN WIDELY PRAISED BOTH INTERSTATE AND INTERNATIONALLY BY SUPPORT GROUPS.

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Supporters

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The Institute thanks all those listed below for their generous financial support during 1 997

JF Gleeson Family Trust Mr & Mrs P Duncan Elmore Apex Club Hughes Fincher Trust Mr Ian Edney Mr & Mrs JR Little JB Were Foundation Mr & Mrs D Heinecke ANZ Trustees Uncle Bobs Club Mr G Heeley Mr & Ms S Skoulis Ms K Behrend Mrs Neilma Gantner Mrs SF Kimpton DrJ M Gooch Mr & Mrs SR Stephenson Mrs R N Cunningham Mrs E M Tallis Mr B R Redpath Mr & Mrs I Bryant Mr R J Edwards The Rousch Family Pacific Publications Mr&Mrs RAGillard Mr DSC Arthur Mrs GA Grimwade Dame Patricia Mackinnon Mr&Mrs LR Mills William Angliss Victoria Charitable Foundation

Mrs J Calvert-Jones Mrs H Handbury Mrs A Kantor Dame Elisabeth Murdoch Calvert-Jones Foundation Mrs J Paterson Miss J Kantor Mr M Kantor The Jack Brockhoff Foundation The Miller Foundation Friends of the Murdoch Institute Mrs J Roxburgh Qld Friedreich's Ataxia Group

Dome Elisabeth Murdoch

Short Statured People Association Professor B Williamson Mrs GE Hale Ms J Fuller Ms K Tabain Klinefelter Support Group of Victoria Professor D Danks Beta Sigma Phi Ballarat Methodist Ladies College (Home Group 9H) ANZ Banking Group Limited

In memory of Keith Troon Mr & Mrs J Bowyer R Aldridge

In memory of David Treseder Mr J Grinter Ms M Wiffen Ms L Spencer Ms A Morell Mr A Barron Ms J Treseder The Giles Family Mr C Gilmer Mr C Stafford Ms C Lamond Ms J Collins Mr & Mrs B Graham Ms G Poultpn «I#:C

Bob Williamson and Lachlan Murdoch

The Kleman Family Mr B Stafford & Ms G Keats The Robins Family Mr & Mrs J Hazen Mr L Rawling Ms K Jackman Mrs T Wright & family Mrs D Armstrong & family Mr & Mrs H Porter Ms W Poyner Mr & Mrs M Robinson The Sinclair Family Mr B Polley MrJ Reid G Chadwick Mr & Mrs M Davis Mr & Mrs K Jones Mr & Mrs 0 Chafey Mr & Mrs R Hood The Beechey Family The Stones Family The McDonald Family Mr & Mrs M Lyons Ms C Gray Ms B Faulmann Mr & Mrs A Page Mrs R M Park Mr & Mrs Hase Mr & Mrs K Spiers Mrs LE Jackson Ms R Desnoy Mrs D I Crabtree

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In Memory of Paul Frueh D E Dyer Ms M Poor Mr & Mrs S Austin V L Thomas Marine Diving Group Mr & Mrs SM Eldredge Ms C Fleet Mr & Mm JR MacLeod Ms MA Lee Mr & Mrs C Braovac

Mr & Mrs T Treseder Mr & Mrs N Gray Mr LA Brown Mr & Mrs Crabtree Mrs J Thorburn Mr F Kostandapoulas Oberon High School Mr & Mrs PJ Cockayne P Thomson Mr & Mrs J McNally Mrs M Ely Mr & Mrs B Treseder

Mrs M Pyman Mr M Brown Mr & Mrs S Clifford Mr & Mrs P Granter Mr AHA Pyman

In Memory of Emma Ritchie J Owens-Brownbill Mr Bob Jenkins Mr & Mrs Fisher Mrs Fraser Ms J Ikin Mr & Mrs L Cupit Mr & Mrs D Haslem Mrs L Louisho & Family Mrs N Beckman Mrs E Brown Mr & Mrs Sutton Mr C Ritchie The Flavell Family Robyna Calisthenics College Police Association Cooperative Credit Mr & Mrs W Ritchie

In Memory of Mitchell Raymond Ross Mr & Mrs G King

In memory of Christopher Arthur Spence The Choy Family Mr & Mrs P Cawthorn Ife iP* Mr B E Griffith Mr & Mrs L Hart Mr & Mrs B Woodward Mr & Mrs A Dickinson Mr JL Swann Mr & Mrs P Lithgow Mr P Shattock Mr & Mrs G Bowden Ms E Deighton Mr & Mrs B Crowe Mr & Mrs C Villeneuve-Smith Mr & Mrs J Ball Mr & Mrs D Evans

In Memory of Shaun Martin

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Ms VA Walsh Mr & Mrs Redman

1 1 In Memory of Matthew Edney

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Mrs BM Barry Mrs K Vinen M M Wightman Mr & Mrs A Drummond Mr & Mrs S Hodgkinson

Mr & Mrs JA Smith Mr & Mrs D Wood The Wood Family Ms R Datsers M Hanton Mr & Mrs Joel S Edelmann Mr & Mrs Eric Edelmann Mr & Mrs SJ Blarney D M Timmins J L Timmins Mr & Mrs D Eakins Mr & Mrs S Keable Mr & Mrs M Burke Mr & Mrs J Caldwell Mr JA Vine PA Robinson D B Kilroy & FM Brown Mr & Mrs C Edney JA Edney Mr & Mrs MP Kulic MrS Giles Mr & Mrs F J Han Ms L Wong Ellis Mastromonaco Jay Jacobsen Mr H Donahue Mr M Renwick Arthur Robinson & Heddiwicks

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Umbrella support groups

Dome Elisabeth and Athol Guy

Support groups for genetic disorders, as the name indi

Operation Jigsaw Report 1997 Operation Jigsaw 1997 was deemed a great success in its first year. This annual education and fundraising effort aims to help research and understanding of those who have inherited disorders such as cystic fibrosis, haemophilia, muscular dystrophy or ataxia. Fundraising projects are closely linked with school curricula giving teachers the opportunity to enrich their science and health programs. All funds raised will benefit the Murdoch Institute, the Melbourne-based Australian Human Genetics Institute, dedicated to improving the lives of children and families with inherited disorders.

Goals

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The impetus for Operation Jigsaw was generated by members of the Friedreich's Ataxia Avssoclafiorj; who, in liaison with the'Murdoch_ |nstitijt&;ctnd’wifi help from St George;'Bartkj began:,Worllhg tovrard two major goals: to’Hncrease community awar%rtess of genetics and genefc 'dis.L^rders; and to raise funds for research into genetic disoVders.

Department of Education

The Department of Education apprbvSf^Wj^d that

Athol Guy, Julie Anthony and Bob Williamson

Media Launch Operation Jigsaw gained media attention when it was officially launched at Parliament House on 30 June 1997. Mr Stephen Elder, Parliamentary Secretary for Education, and Dame Elisabeth Murdoch, patron of the Murdoch Institute spoke at the launch as did Mr Athol Guy, St George Bank, and Professor Bob Williamson. Channel 9 Health Reporter, Belinda Byrne, featured the launch on the Channel 9 News.

cates,

PROVIDE SUPPORT AND INFORMATION FOR OTHERS WITH A SIMILAR CONDITION. ThE GROUPS ARE A VITAL LINK BETWEEN THE SUPPORT AND FAMILY MEMBERS AND MEDICAL PROFESSIONALS INFORMATION ARE PROVIDED BY THE GROUPS, AND IDEAS ARE EXCHANGED. SUPPORT GROUPS HAVE BEEN A RECOGNISED AND VALUED PART OF THE CLINICAL GENETICS SERVICE AT THE VCGS FOR OVER TEN YEARS.

The Department of Human Services' 1997 discussion paper, "Genetic Services in Victoria", endorsed the formation of AN Umbrella Support Group. This group would provide community

participation

its

representation

Services

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Sponsor and Prize Operation Jigsaw is indeed grateful to St George Foundation for their generous contribution of $30,000 to cover set-up expenses and prizes for 1997. The student prize of $2,000 was awarded to Jonathan Zimet, a Grade 5 student at Bialik College, Havsrthorn. Jonathan's school also received $2,000.

Plans culminated in December 1997 when the Umbrella Group concept was launched with over 20 groups repre sented. A working group has now been formed with the Julie Hoy and the Department of Human Services, Margaret Sahhar and Maureen Crawford (VCGS), and Nola Horne (a Genetic Counselling student and also a

Human

Victorian Department Advisory Committee.

Genetics

CONSULTANT WITH THE DEPARTMENT OF HUMAN SERVICES).

1998 Operation Jigsaw 1997 has provided a solid founda tion for a more substantial effort, both in schools and in fundraising in 1998. Many thanks to those who gave their time, effort and money in 1997. For information on how to be a part of Operation Jigsaw 1998, telephone the Murdoch Institute on (03) 9345 5045 or write to:

Aims of the working group are: TO FORM AND NAME THE UMBRELLA GROUP TO PROVIDE RESOURCES TO ALL THE MEMBER GROUPS TO SELECT A REPRESENTATIVE TO ATTEND THE GENETICS

Advisory Committee meetings TO PROVIDE CONSUMER FEEDBACK TO THE VCGS AND THE

Murdoch Institute Operation Jigsaw, Murdoch Institute, 1 0th Floor, Royal Children's FIospital, Flemington Road, Parkville VIC 3052

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Friends of the Murdoch

M.l. 7 group and Bob Williamson

After an extremely busy Year in 1996 with our Christmas Trees festival, we are indebted to our President Ann McFarling and also Elizabeth and Michael Richards for opening their gardens for our 1997 fundraisers. As part of the Open Garden scheme, the beautiful gardens in Malvern and Armadale were opened to the public and proceeds were donated to the Institute. Funds were boosted by the sale of homemade preserves, handmade Christmas gifts, iris plants and Murdoch Institute Christmas cards.

M.l. 7 and the First Shelley Beach Classic The M.l. 7 is a newly formed group with a sporty theme for its fundraising agenda. 1997 was spent organising our first fun day, a lunch and tennis round robin played on courts in the Portsea area in February,!998. In magnificent weather our tennis was played and afterwards we converged on the magnifent lawns of the Calvert-Jones' beach house, "Flarrodene". It was time to relax with friends and participants, draw the raffle and present the prizes. Proceeds from the day will go towards the communi ty education program of the Murdoch Institute and the VCGS. Plans for the 1999 Shelley Beach Classic Tennis day are underway.

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Scientific Officers and Research Assistants

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Murdoch Institute Staff 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., M.B.A.

Senior Scientists Jim Camakaris, B.Sc.(Hons.), Ph.D. K.H. Andy Choo, B.Sc.(Hons.), Ph.D. Hans-Henrik Dahl, Lie. scient. (Denmark), Ph.D. Susan Forrest, B.Sc.(Hons.), D.Phil.(Oxon.), B. Bus. (Bus. Admin.) i Jane Halliday, B.Sc.(Hons,)/ Bh.t Panos loannou, B; Sc.(Hons.), Ph Julian Mercer, Bf|ic.(Hons.), ThT Donald Newgreen, B.Sc.(Hons.), David Thorburn, B.Sc.(Hon^,), Ph.D.*^,V '

Institute Visitors ^ Brandon Wainwright (Queensland^ B.Sc.(Hons.), Bendicht Wermuth!|$witzerland),

Clinical Scientists Agnes Bankier, M.B., B.S., F.R.A.C.P. Avihu Boneh, M.D., Ph.D. Maureen Cleary, M.B. ChB. M.R.C.P. (UK) Mac Gardner, M.B., Ch.B., M.Sc., F.C.C.M.G., 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., F.R.A.C.P. Johan Van Hove, M.D., Ph.D.

Postdoctoral Fellows Michael Cancilla. B.Sc.(Hons.), Ph.D. Suzi Cutts, B.Sc.(Hons.), Ph.D. Kate Elliott, B.Sc.(Hons.), Ph.D. Tracy Evans-Whipp, B.Sc. (Hons.), Ph.D. Peter Farlie, B.Sc.(Hons), Ph.D. Sharon La Fontaine, B.Sc. (Hons.), Ph.D. Richard Saffery, B.Sc. (Hons.), Ph.D. Daniel Strausak, Lie. phil. nat. (Switz.), Ph.D. Stephen Wilcox, B.Sc. (Hons.), Ph.D. Kathy Williamson, B.Sc. (Hons.), Ph.D.

Education Unit MaryAnne Aitken, B.Sc.(Hons.), Ph.D., Grad.Dip.Genetic Counselling, S.R.N. Mark Fisher, B. Ed. Sylvia;Metcalfe, B.Sc.(Hons.), Ph.D.

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Stuart Beattie, B.Sc. (Hons) Dianne Beck, B.Appl.Sci.(App.Biol.) Ami Ben-Yaacov, Visiting Scientist (Israel) Hilary Brooks, B.Sc. (Hons.) Veronica Collins, B.Sc., Grad.Dip.Ed., M.Sc. Sherry Cook, B.Sc.(Hons.) Marjorie Crawford, A.R.M.l.T. Elizabeth Earle, A.A.I.M.L.S. Stephen Firth, B.Sc., M.App.Sci. Elizabeth Fitzpatrick, B.Sc.(Hons.) Kerry Fowler, M.App.Sci., M.Sc., Grad.Dip.Ed. Fanne Galjaard, Visiting Medical Student (Holland) Andrew Grimes, B.App.Sci. Linda Hii, B.Sc., M.Sc. Joanne Hill, B.App.Sci. Wendy Hutchison, B.App.Sci.(App.Biol.) Danielle In/ine, B.Sc. (Hons.) Denise Kirby, B.Sc.(Hons.) Melanie Knight, B.Sc.(Hons.) Geraldine McDonald, S.R.N., Grad.Dip.Wom.Health, M.P.H. Lachlan McDonald, B.Sc.(Hons.) Henny Miller Joseph Minichiello, B.Sc.(Hons.), M.Sc. Rachel O'Dowd, B.Ag.Sci. Erin Oldaker, B.App.Sci. Amelia Osborn, B.App.Sci. Damien Paris, B.Sc. (Hons.) Janet Shaw, B.Sc. (Hons.) Tiffany Symes, B.Sc.(Hons.) Kellie Tainton, B.Sc. (Hons.) Marietta Veldman, Visiting Medical Student (Holland) Rosemary Warren, S.R.N., R.M., G.A., Grad.Dip.Soc. Carole Webley, B.Sc., Grad.Dip.Gomputing;; M P-H.

Ph.D. Scfiolars

Damien Hudson, B.Scj ^^^^litsis, B.Sc. 7 Ken;, B.Sc;-(Hons.) Anthony Lo, B.Tyled.S'c., M.B., Ch.B Paul Lockhart, B.Sc. (Hons.) Andrew M(igDonald/ BvSiifr(H6ns.) . Kumaran Nprayanan, B.Sc. (Ho Laraine Peters, B.Sc.(Hons.) Michael f|etris, B.Sc.(Hons.) Swari Setfeningsih, (y).D. Michael l|heophilos) BiSc.(Hons.) Tom Van lAgtrnael, M.Sc. ■, Louise Wdngerek, B.Ag.Sci. r ' Sarah White, B.Sc. (Hons.^

Technical Assistants Zoe Anderson, Assoc.Dip.App.Sci. Roseanna Bhagwandas Alison Blake, Cert.App.Sc. Robyn Breslin Blanche Dekker Sophie Gazeas, Assoc.Dip.App.Sci.(Animal Technology) Michelle Guneratne Karyne Lord Matthew Newman, Assoc.Dip.Lab.Techniques Amanda Notini Rosario Reyes, Assoc.Dip.App.Sci.(Lab.Technology) Anick Sylvain

Dietitian Dorothy Francis, SRD Administration Accountant: Viren Abeyasinghe, M.B.A., F.C.M.A. Personnel Assistant: Debbie Zombolas Personal Assistant to the Director: Julie Foletta, B.

Ed. Administrative Assistants

Katie Allen, 6.Med.Sc., M.B., B.S. Loreta Ambrosini, B.App. Sci.(Hons.) Alyssa Barry, B.Sc. (Hons.) Martin Delatycki, M.B., B.S., F.R.A.C.P. Jan Fullerton, B.Sc.(Hons.) Catherine Hearn, B.Sc. (Hons.) Michelle Howie B.Sc.(Hons.) Emily Howman, B.Med. Lab. Sci

Debbie Davis Vicki Hirt Fiona Keltie Colleen King Kristine Yeomans

Photography/Graphic Design Michele Winsor Voula Pashalidis

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David Francis, B.Sc.(Hons.), M.Sc. Lee Harrison, B.Sc.(Hons.) Louise Hills, B.Sc. Sara Nouri, B.Sc. (Hons.) Ralph Oertel, B.Sc., H.G.S.A.A.C. Vida Petrovic, B.Sc., H.G.S.A.C.C. Vladimir Pupko, B.Sc.Agr. Anne Robertson, B.Sc., H.G.S.A.C.C. Howard Slater, B.Sc., Ph.D., Dip.R.C.Path., H.G.S.A.C.C. - Scientist in charge Marie Thorpe, B.Sc.(Hons.), Dip.Ed. Cathryn Vaux, B.Sc., Grad. Dip. Genetic Counselling, H.G.S.A.A.C. Lucille Voullaire, M.Sc., H.S.S.A.C.C. Lorna Webber, T.P.T.C., B.Sc. (Hons.), Ph.D.

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Victorian Clinical Genetics Services Executive Director Robert Williamson, Ph.D.,FRCPath., Hon. MRCP, Hon.M.D. (Turku)

Metabolic Physicians Avihu Boneh, M.D., Ph.D., Maureen Cleary) M-B., Ch.B^j

Clinical Fellows Karen Dunn, B.Med.Sci. M)B., B.tSimon Hauser, M.B., B.S., F.R.A.C. Helen Heussler, M.B., B.S., F.R.A.C

Heidi Peters, M.B., B.S., FVfciC.P. Stephen Robertson* M.B., Ch.B., F.R.A. Catherine Rose, M.B., B.S. Ravi Savarirayan, M.B., B S , F RvA, C.P.

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DNA Diagnosis - Scientists

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Ivan Biros, B.Sc., Ph.D. Janice Brasch, B.Sc.(Hons.), M.Sc. Jenny Douglas, B.Sc. (Hons.) Susan Forrest, B.Sc.(Hons.), D.Phil.(Oxon.), B.Bus.(Bus.Admin.) - Scientist in charge Karina Forshaw, B.App.Sci. Kathy Garofalo, (Trainee) Jean Hendy, B.Sc., F.A.I.M.L.S. Steven Nasioulas, B.Sc. (Hons.) Andrea Twomey, B.Sc.(Hons.)

Sue Casanelia, S.R.N. Susan Clement, M.Sc., B.A. Susan Fawcett, B.Sc., Grad.Dip.Genetic Counselling Clara Gaff, B.Sc.(Hons.), Ph.D. Margaret Olsen, Dip.App.BioL, Dip. Ed. Ann Robertson, S.R.N. Linda Warwick, S.R.N. Jo Wells Mary-AnneYoung, S.R.N.

Cytogenetics - Scientists

Robin Forbqs

Selena Bourke, B.Sc. (Hons.) Trent Burgess, B.Sc.(Hons.) Melissa Curtis, B.Sc. Sue Dale, B.Sc.(Hons.) Julie Davies, B.Sc., H.G.S.AiA.C. Desiree du Sart, B.Sc., Ph.D. Tracy Fleming, B.Sc. (Hons.)

f Social Workers

Business Manager Anne Cronin, B.Sc., B.Bus.(Acc.), A.S.A., C.P.A.

Laboratory Manager

Esilio Benedetti Ian Brooks Wall Drummond, Dip. Basic Med. Sci. Fernando Garcia Bozena Jezierski, Dip. Ed. Thomas Milovac, Assoc. Dip. App. Sci. (Lab. Tech.)

Lynda Phillips I. Project Officer Maternal Serum Screening

Genetic Counsellors / Genetic Clinic Co-ordinators

Leonard Bonacquisto, B.Sc.(Hons.), Dip.Comp.Sci. Mona El-Masri Ivan Francis, B.Sc., Dip.Comp.Sci. - Scientist-inCharge Manal Ibrahim, B.Sc. Nella Napolitano, B.App.Sci.(Med.Lab.Sci.) Nick Tzanakos, B.App.Chem.

Technical Assistants

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Screening Laboratory - Scientists

Clinical Geneticists Agnes Bankier, M.B., B.S., F.R.A.C.P. Martin Delatycki, M.B., B.S., F.R.A.C.P. Mac Gardner, M.B., Ch.B., M.Sc., F.C.C.M.G., 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., F.R.A.C.P.

Barry Holt, B.App.Sci.(M.T.), A.A.I.M.L.S., M.B.A.

Administrative Assistants Christine Keenan Michele Mourik Sharon Vandersluis Lorraine White

Computer Support Shilpa Shah

Susan Mansie, S.R.N., B.S.W. Margaret Sahhar, B.A., Dip.Soc. Studies Maureen Crawford, B.A., Dip.Soc.Studies, Dip.Ed.

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Published and accepted for publication since 1996 report Ackland, LM, J Cornish, JA Paynter, A Grimes, A Michaiczyk, and JFB Mercer. Expression of Menkes (ATP7a) and Wilson (ATP7a) disease genes in mam mary carcinoma cells. Biochem. J. 328: 237-243, 1997. Allen, K and PF Whitington. "Evaluation of liver func tion." In Fetal and Neonatal Physiology, ed. R Polin and W Fox. 2. Philadelphia: W.B. Saunders, 153452, 1997.

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Bines, J, D Francis, and D Hill. Reducing parenteral requirement in children with short bowel syndrome; Impact of an amino acid-based complete infant for mula. Journal of Pediatric Gastroenterology and Nutrition 26: 123-128, (in press).

Biros, I, K Forshaw, U Sheffield, AJ Kornberg, and S Forrest. Spinal muscular atrophies - an ongoing diag nostic dilema? Molecular Genetics (in press). Blok, R.B., D.A. Gook, D.R. Thorburn, and H-H.M. Dahl. Skewed segregation of the mtDNA nt8993 (T>G) mutation in human oocytes. Am J Hum Genet 60: 1495-1501, 1997.

Allen, KJ, EB Rand, J Hart, and PF Whitington. Prognostic implications of centrilobular necrosis in pediatric hepatic allograft recipients. Transplantation 65: 692-8, (in press).

Concilia, MR, J Graves, LE Matesic, RH Reeves, KM Tainton, KHA Choo, MA Resnick, VL Larionov, and NY Kouprina. Rapid cloning of mouse DNA as yeast artificial chromosomes by transformation-associated recombination (TAR). Mammalian Genome 9: 157159, (in press).

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).

Concilia, MR, KM Tainton, AE Barry, V Larionov, N Kouprina, MA Resnick, D du Sort, and KH Andy Choo. Direct cloning of human 10q25 neocen tromere DNA using transformation-associated recombination (TAR) in yeast. Genomics 47: 399404, (in press).

Bankier, A. "Practical Paediatrics." In 4th Edition, Chapter 6, Approach to the Dysmorphic Child, ed. M Robinson and D Roberton. Churchill Livingstone, 1997.

Cashman, JR, YA Bi, J Lin, R Youil, M Knight, S Forrest, and E Treacy. Human flavin-containing monooxygenase form 8; cDNA expression of the enzymes containing amino acid substitutions observed in individuals with trimethylaminuria. Chemical Research in Toxicology 10: 837-841, 1997.

Beattie, JH, AM Wood, AM Newman, I Bremner, KHA Choo, AE Michalska, JS Duncan, and P Trayhurn. Characterisation of metallothionein (-1 and -II) null mice. MT-97 Proceedings (in press). Beattie, JH, AM Wood, AM Newman, I Bremner, KHA Choo, AE Michalska, JS Duncan, and P Trayhurn. Obesity and hyperleptinaemia in metallothionein (-1 and -II) null mice. Proc. Natl. Acad. Sci. 95: 358363, (in press).

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EE Grqen, B Horsthemke, R Williamson, C Huxley, cfid C Coutelle. Isolation of DNA from the centromere of human chromosome 7 by microdissiction. Chromosome Research (in press). |

Choo, KHA. The centromere. Oxford University Press: 1-304, 1997.

Choo, KHA. Centromere DNA dynamics: Latent cen tromeres and neocentromere formation. American Journal Human Genetics 61: 1225-1233, 1997. Choo, KHA. Human artificial chromosomes and gene therapy. Today's Life Science 9: 14-18, 1997. Choo, KHA. Turning on the centromere (news and views). Nature Genetics 18: 3-4, (in press). Choo, KHA. Why is the centromere so cold? (Insight/Outlook). Genome Res. 8: 81-82, (in press). Cleary, MA, DEM Francis, and NM Kilpatrick. Oral health implications in children with inborn errors of intermediary metabolism: a review. International Journal of Paediatric Dentistry 7: 133-141, 1997. Collins, VR, C Webley, U Sheffield, and JL Halliday. Fetal outcome and maternal morbidity after early amniocentesis. Prenat. Diagn. (in press). Cotton, RGH, E Edkins, and S Forrest. Mutation detection: a practical approach. IRL Press (in press).

Dahl, HHM and W Hutchison. Analysis of in vivo DNA methylation. Methods in molecular biology, (in press).

ICQ Dong, Y, SJ Berners-Price, DR Thorburn, T Antalis, J Dickinson, T Hurst, L Qiu, SK Khoo, and PG Parsons. Serine protease inhibition and mitochondrial dysfunc tion associated with cisplatin resistance in human tumor cell lines: targets for therapy. Biochem. Pharmacol. 53: 1673-1682, 1997. Dowsing, B, A Puche, C Hearn, and Key B. The pres ence of novel N-CAM glycoforms in the rat olfactory system. J. Neurobiol. 32: 659-670, 1997.

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 gastrointestinal features: X linked or mitochondr ial aetiology? J. Med. Genet. 34: 520-524, 1997. Delatycki, M and RJM Gardner. Three cases of tri somy 13 mosaicism and a review of the literature. Clin. Genet. 51: 403-407, (in press). Delatycki, MB, D Paris, RJ McKinlay Gardner, K Forshaw, GA Nicholson, N Nassis, R Williamson, and SM Forrest. Sperm DNA analysis in a Friedreich atax ia premutation carrier suggests both meiotic and mitotic expansion in the FRDA gene. J. Med. Genet, (in press). •5^^ —. Pe^el and R.J.M. Gardner. Delatycki, M., Trisomy 13 mospicism at prenatal diagnosis: dilemmas in in' ' " ' ’ ' s... - -49, (in press).

Delatycki, M and JG Rogers. "The genetics of fibrodysplasia ossificans progressiva." In Fibrodysplasia ossificans progressiva. Clinical orthopaedics and related research., ed. FS Kaplan. 346. 15-18, 1997. 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 SuppI 1: 5314-S316, (in press). Delatycki, M and LJ Sheffield. Familial heterotaxia: What is the inheritance in this family? Am. J. Med. Genet. 69: 429-430, 1997. Denoyelle, F, D Weil, MA Maw, SA Wilcox, NJ Lench, DR Allen-Powell, AH Osborn, HHM Dahl, A Middleton, MJ Houseman, C Dode, S Marlin, A Boulila-EIGaied, M Grati, H Ayadi, 5 BenArab, P Bitoun, G Lina-Granade, J Godet, M Mustapha, J Loiselet, E El-Zir, A Aubois, A Joannard, J Levilliers, EN Garabedian, RF Mueller, RJM Gardner, and C Petit. Prelingual deafness: high prevalence of a 30delG mutation in the connexin 26 gene. Human Molecular Genetics 6: 2173-21 77, 1 997, Dierick, H, JFB Mercer, and T Glover. A phpsphoglycerate mutase brain isoform (PGAM 1) pseudo gene is localized within the Menkes diseasei gene (ATP7A). Gene 198: 37-41, 1997. *

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du Sort, D, MR Concilia, E Earle, J-i Mao, R Saffery, KM Tainton, P Kalitsis, J Martyn, AE Barry, and KH Andy Choo. A functional neo-centromere formed through activation of a latent human centromere and consisting of non-alpha-satellite DMA. Nature Genet. 16: 144-153, 1997. du Sort, D. and K.H.A. Choo. "Simultaneous fluores cence in situ hybridisation and anti-centromere anti body staining of normal and "stretched" metaphase chromosomes." In Molecular Biotechniques, ed. R. Rapley and J.M. Walker. Totowa, NJ, USA: Humana Press, (in press).

Gates, PC, D Paris, SM Forrest, R Williamson, and RJM Gardner. Friedreich's ataxia presenting as adult onset spastic paraparesis. Neurogenetics (in press), 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 CYPlla with polycystic ovary syndrome and hyperandrogenism. Hum. Mol. Genet. 6: 397-402, 1997.

Fowler, KJ, AJ Newson, AC MacDonald, P Kalitsis, MS Lyu, CA Kozak, and KHA Choo. Chromosomal localisation of mouse CenpAgene. Cyto. & Cell Genet, (in press).

Giunta, C., R. Youil, D. Venter, C.W. Chow, G. Somers, A. Lafferty, B. Kemper, and R.G.H. Cotton. Rapid diagnosis of germline p53 mutation using the enzyme mismatch cleavage method. Diagn. Mol. Path, (in press).

Franks, S, N Gharani, D Waterworth, S Batty, D White, R Williamson, and M McCarthy. Current developments in the molecular genetics of polycystic ovary syndrome. Trends in Endocrinology and Metabolism (in press).

Grimes, A., J. Paynter, I.D. Walker, M. Bhave, and J.F.B. Mercer. Reduction of carbonic anhydrase III in the liver of the mouse mutant "toxic milk" due to cop per accumulation. Biochem J 321: 341-346, 1997.

Gardner, RJM and JJ Tjandra. "Familial cancer due to predisposing genes." In Textbook of Surgery, ed. G.J.A. Clunie, J.J. Tjandra, and D.M.A. Francis. Blackwell, 1997.

Grimes, A, CJ Hearn, P Lockhart, DF Newgreen, and JFB Mercer. Molecular basis of the brindled mouse mutant (mobr): a murine model of menkes disease. Human Molecular Genetics 6: 103-1042, 1997. Grimes, A, C Hearn, P Lockhart, D Newgreen, and JFB Mercer. Molecular basis of the brindled mouse mutant (Mobr): a murine model of Menkes disease. Human Molecular Genetics 6: 1032-1042, 1997.

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Kedinger, M. and D. Newgreen. "The gut and enteric nervous system." In In: Birth Defects: Perspectives from Contemporary Development Biology, ed. P Thorogood (ed). UK: John Wiley & Sons, 153-196, 1997. Kerr, RSE and DF Newgreen. Isolation and character ization of chondroitin sulfate proteoglycans from embryonic quail that influence neural crest cell behaviour. Developmental Biology 192: 108-124, 1997.

m Hall, RK, A Bankier, MJ Aldred , K Kan, JO Lucas, and AGB Perks. Solitary median maxillary central incisor, short stature, choanal atresia/midnasal steno sis (SMMCI) syndrome. Oral Surg. Oral Med. 84: 651-662, 1997. Halliday, JL, O Griffin, A Bankier, C Rose, and M Riley. Use of record-linkage between a statewide genetics service and a birth defects/congenital mal formations register to determine use of genetic coun selling services. Am. J. Med. Genet. 72: 3-10, 1997. Hauser, S. Autoimmune (idiopathic) thrombocy topenic purpura. The Lancet 350: 368, 1997. Hauser, SF, JM Chemke, and A Bankier. Pelvis-shoul der dysplasia. Ped. Radiology (in press). Hauser, S and H Peters. Glutaric aciduria type 1: An underdiagnosed cause of encephalopathy and dysto nia-dyskinesia syndrome in children, (in press) n Hudson, D, KL Fowler, F Fade, R Saffery, P Kalitsis, H Trowell, J Hill, ND Wreford, DM de Kretser, MR Concilia, F Howman, L Hii, SM Cutts, DV Irvine, and KHA Choo. Centromere protein B null mice are mitotically and meiotically normal but have lower body and testis weights. J. Cell Biol, (in press).

lannello, R.C., J. Young, S. Sumarsono, M. Tymms, H.-H.M. Dahl, and I. Kola. Regulation of PDHa-2 expression is mediated by proximal promoter sequences and CpG methylation. Mol Cell Biol 17: 612-619, 1997. Ikeda, H, Y Matsubara, H Mikami, S Kure, M Owada, T Gough, PM Smooker, M Dobbs, HHM Dahl, RGH Cotton, and K Narisawa. Molecular analysis of dihydropteridine reductase deficiency: identification of two novel mutations in Japanese patients. Human Genetics 100: 637-642, 1997. Kalitsis, P, KJ Fowler, E Earle, J Hill, and KHA Choo. Targeted disruption of mouse centromere protein C gene leads to mitotic disarray and early embryo death. Proc. Natl. Acad. Sci. 95: 1136-1141, (in press). Kalitsis, P, AC MacDonald, AJ Newson, DF Hudson, and KHA Choo. Gene structure and sequence analy sis of mouse centromere proteins A and C. Genomics 47: 108-114, (in press). Kaplan, FS, M Delatycki, FH Gannon, JG Rogers, and R Smith. "Fibrodysplasia ossificans progressiva." In Inherited neuromuscular disorders: Recent advances & future prospects, ed. A.E.H. Emery, (in press). J

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Kouprina, N, J Graves, MR Cancilla, MA Resnick, and V Larionov. Specific isolation of human rDNA genes by TAR cloning. Gene 197: 269-276, 1997. La Fontaine, S, SD Firth, P Lockhart, JA Paynter, and JFB Mercer. Low copy number plasmid vectors for eukaryotic gene expression: transient expression of the Menkes protein. Plasmid (in press). Lagerstrom-Fermer, M, M Sundvall, F Johnsen, GL Warne, SM Forrest, JD Zajac, A Rickards, D Ravine, U Landegren, and U Pettersson. X-Linked recessive panhypopituitarism associated with a regional dupli cation in Xq25-q26. American Journal of Fluman Genetics 60: 910-916, 1997. Lamande, SR, JF Bateman, W Hutchison, RJM Gardner, SP Bower, F Byrne, and H-HM Dahl. Reduced collagen VI causes bethlem myopathy: a heterozygous COL6A1 nonsense mutation results in mRNA decay and functional haploinsufficiency. Hum. Mol. Genet, (in press). Lennox, N, J Cohen, H Slater, and A Cook. The frag ile X syndrome. Australian Family Physcian 27: 163166, (in press).

Macreadie, IG, DR Thorburn, DM Kirby, LA Castelli, NL de Rozario, and AA Azad. HIV-1 protein Vpr caus es gross mitochondrial dysfunction in the yeast Saccharomyces cerevisiae. FFBS Letters 410: 145149, 1997. Mercer, JFB. Gene Regulation by copper and the basis for copper homeostasis. Nutrition 13: 48-49, 1997. Mercer, JFB. Genetic disorders of copper metabolism and the dual nature of copper in biology in "Copper" National Environmental Health Forum Monographs, Metal Series No.3. Edited by M.R. Moore, P. Imray, C. Dameron, P. Callan, A. Langley and S. Mangas 16-30, 1997. Mercer, JFB. Menkes disease and animal models. Am. J. Clin. Nutrition (in press). Mercer, JFB and DM Danks. "Disorders of copper transport." In update to chapter 68 CD ROM version Metabolic and molecular basis of inherited disease., ed. Al Baudet CR Scriver, WM Sly and D. Valle. New York: McGraw-Hill, Version 1, (1997). Mercer, J.F.B. and J. Camakaris. "Inherited disorders of copper transport in mammalian systems." In Metal Ions in Gene Regulation, ed. Chapman and Hall S. Silver and W. Walden. New York: 250-276, 1997.

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Robertson, S, K O'Doy, and A Bankier. The 4q- syn drome; further delineation of the minimal critical region. Clin Genet (in press. Robertson, S, H Lipp, and A Bankier. Zimmerman Laband syndrome in an adult. Long term followup of a patient with vascular and cardiac complications. Am J Med Genet (in press). Robertson, S, T Gunn, B Allen, C Chapman, and D Becroft. Are Melnick-Needles syndrome and otopalato-digital syndrome type II allelic? American Journal of Medical Genetics 71: 341-347, 1997.

Moores, C., J.G. Rogers, I.M. McKenzie, and T.C.K. Brown. Anaesthesia for children with mucopolysac charidoses. Anaesth Intens Care 24: 459-463, 1996. Nasioulas, S., L. Sheffield, S. Mansie, and S. Forrest. Modified protocol for the detection of the CAG repeat expansion in Huntington's disease and appli cation to a predictive testing protocol. Mol. Diag. (in press). Newgreen, D.F., B. Southwell, L. Hartley, and I.J. Allan. Migration of enteric neural crest cells in rela tion to the growth of the embryonic avian gut. Acta Anat 157: 105-115, 1997. Newgreen, DF, RS Kerr, J Minichiello, and N Warren. Changes in cell adhesion and extracellular matrix molecules in spontaneous spinal neural tube defects in avian embryovTeratology 55: 195-207, .1997.

. Ogle, R.F., J. Christodoulou, E. Fagan, R.^ Blok, D.M. Kirby, K.L. Seller, H.-H.M. Dahl, and D.R. Thorburn. Mitochondrial myopathy with tRhjA Leu (UUR) mutation and complex I deficiency responsive to riboflavin. J. Paed. 130; 138-145, 1997. |

Rogers, JG. "Practical paediatrics." In 4th Edition: Chapter 7: Genetic counselling, ed. D. Roberton, M. Robinson, (in press). O'Neill, M, W Brewer, C Thornley, D Copolov, G Warne, A Sinclair, S Forrest, and R Williamson. The kallmann syndrome gene (KAL-X) is not mutated in schizophrenia. American Journal of Medical Genetics (in press). Peters, HL and A Bankier. Lipomatous myelomeningocoele, athyrotic hypothyroidism, sensorineural deafness: a new form of syndromal deaf ness? (in press). Riley, BP and R Williamson. Non-parametric analysis of chromosome 6p24-22 marker data and schizo phrenia in southern African Bantu-speaking families. Psychiatric Genetics 7: 131-132, 1997.

Robertson, SP, GL Klug, and JG Rogers. Cerebirospinal fluid shunts in San Filippo syn|Jrome (MPS jll). Eur. J. Pediatrics (in press).

-V.-

. ill * '-'tAIji I

r,r

Robertson, S, B Tsang, and S Aftimos. Cerebral infarction in Noonan syndrome. American Journal of Medical Genetics 71: 111-114, 1 997.

Robertson, S and D Simmons. Determinants of infant adiposity in ethnic groups with a high prevalence of NIDD^a prospective study,.Diqb Res Clin Pract. (in press)' AS,

-w,

Rogers, JG and J Halliday. "Genetic counselling, antenatal diagnosis, serum and folate supplementa tions." In Orthopaedic Management of Spina Bifida (3rd Edition), ed. M.B. Menelaus. (in press). 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).

Savarirayan, R and A Bankier. Acampomelic cam pomelic dysplasia with de novo 5q;17q reciprocal translocation and sever phenotype. Journal of Medical Genetics (in press). Savarirayan, R, R Couper, J Nance, L Morris, and E Haan. Osteopathia striata with cranial sclerosis: Highly variable phenotypic expression in five family members spanning four generations. American Journal Human Genetics 61: A111, 1997. Savarirayan, R, P Tomlinson, and EM Thompson. Baller-gerold syndrome associated with congenital portal venous malformation. Journal Medical Genetics (in Press). Setianingsih, I, R Williamson, S Marzuk, A Harahap, M Tamam, and S Forrest. Molecular basis of betathalassemia in Indonesia: Application to prenatal diagnosis. Molecular Diagnosis (in press). Sheffield, U, AH Osborn, WM Hutchison, DO Sillence, Sm Forrest , SJ White, and HHM Dahl. Segregation of mutations in arylsulfatase E and cor relation with the clinical presentation of chondrodys plasia punctata. Medical Genetics (in press).

Waterworth, DM, ST Bennett, N Gharani, Ml McCarthy, S Hague, S Batty, GS Conway, D White, JA Todd, S Franks, and R Williamson. Linkage and association of insulin gene VNTR regulatory polymor phism with polycystic ovary syndrome. The Lancet 349: 986-990, 1997. Webley, C and J Halliday. Report on prenatal diag nostic testing in Victoria, 1996. Murdoch Institute ISSN 1327-7618, 1997. Shore, EM, JG Rogers, R Smith, FH Gannon, M Delatycki, JM Connor, JA Urtizberea, J Triffitt, M Le Merrer, and FS Kaplan. "Fibrodysplasia ossificans progressiva." In The genetic aspects of osteoporosis and metabolic bone disease, ed. M.J. Econs. Simmons, D and S Robertson. Influence of material insulin treatment on the infants of women with gesta tional diabetes. Diabetic Medicine 14: 762-765, 1997. Takayama, K., D.M. Danks, E.P. Salazar, J.E. Cleaver, and C.A. Weber. DNA repair characteristics and mutations in the ERCC2 DNA repair and tran scription gene in a trichothiodystrophy patient. Hum Mut (in press).

Thompson, GN, BYL Hsu, JJ Pitt, E Treacy, and CA Stanley. Fasting hypoketotic coma in a child with defi ciency of mitochondrial 3-hydroxy-3-methylglutarylCoA synthase. The New England Journal of Medicine 337: 1203-1207, 1997. Thompson, G.N. "In vivo stable isotope methods in inborn errors of metabolism." In Advances in chemi cal diagnosis and treatment of metabolic diseases, Vol 2, 2. New York: Wiley, (in press). Toder, R, J Wienberg, L Voullaire, PCM O'Brien, P Maccarone, and JAM Graves. Shared DNA sequences between the X and Y chromosome in the tammar wallaby - evidence for independent additions to eutherian and marsupial sex chromosomes. Chromosoma 106: 94-98, 1997. Treacy, EP, BR Akerman, R Youil, C Bibeau, J Lin, AG Bruce, M Knight, DM Danks, JR Cashman, and SM Forrest. Mutations of the flavin-containing monooxy genase gene (FM03) cause trimethylaminuria, a defect in detoxification. Human Molecular Genetics (in press). Trent, RJ, LJ Sheffield, Z-M Deng, WS Kim, NT flassif, C RyeCG Woods, RC Michaelis, J Tarleton, and A Smith The elusive Angelman syndrome .critical regiori. J. Med. Genet. 34: 714-718, 1997.

Wexler, ID, SG Hemalatha, J McConnell, NRM Buist, HHM Dahl, SA Berry, SD Cederbaum, MD Patel, and DS Kerr. Outcome of pyruvate dehydrogenase defi ciency treated with ketogenic diets: studies in patients with identical mutations. Neurol. 49: 1655-1661, 1997. White, SL, DR Thornburn, J Christodoulou, and Dahl HHM. A novel mtDNA variant that may give a false positive diagnosis for the T8993C mutation. Molecular Diagnosis (in press). Wildenauer, DB, R Williamson, and and others in the consortium. Additional support for schizophrenia link age on chromosomes 6 and 8: A multicenter study. American Journal of Medical Genetics 67: 580-594, 1996.

Williamson, R. Detecting fetal abnormalities. Australian Family Physician 26: 245-253, 1997. Williamson, R. Healthy genes in a healthy body. WHO (in press). Williamson, R. Gene therapy. Australian Prescriber 20: 72-74, 1997. Yang, Z, R Youil, D Thorburn, CW Chow, RGH Cotton, and GS Baldwin. Absence of common tri functional protein mutation in patients with Alpers dis ease. J InherMetab Dis 20: 828-830, 1997. Youil, R., S.J. Ramus, and R.G.H. Cotton. "Comparison of mutation detection of chemical cleavage of mismafch (CCM) and enzyme mismatch cleavage." In Laboratory methods for the detection of mutations and polymorphisms in DNA., ed. GR Taylor. Baton Rouge: CRC Press, 239-253, 1997. Young, H., D. Ciampoli, B. Southwell, and D.F. Newgreen. Origin of the interstitial cells of Cajal in the mouse intestine. Dev Biol 180: 97-107, 1996.

Balance Sheets

■Inancial Reportj

Consolidated 1997 1996 $ $

i01st December I 1997

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The Company 1997 1996 $ $

CURRENT ASSETS Cash Receivables Investments TOTAL CURRENT ASSETS

1,202,917 1,197,580 2,048,955 4,449,452

397,450 1,125,685 2,205,028 3,728,163

713,612 306,711 1,876,535 2,896,858

375,749 371,547 2,036,439 2,783,735

NON-CURRENT ASSETS Receivables Investments Property, Plant & Equipment TOTAL NON-CURRENT ASSETS

870,000 13,292,470 1,609,277 15,771,747

942,500 10,347,524 1,269,739 12,559,763

870,000 13,189,766 1,512,002 15,571,768

942,500 10,244,820 1,216,080 12,403,400

TOTAL ASSETS

20,221,199

16,287,926

18,468,626

15,187,135

CURRENT LIABILITIES Accounts Payable Provisions TOTAL CURRENT LIABILITIES

1,636,922 1,085,613 2,722,535

976,942 955,215 1,932,157

735,348 521,868 1,257,216

622,661 472,396 1,095,057

98,931 232,285 331,216

139,191 162,874 302,065

71,854 83,367 155,221

85,332 56,240 141,572

The Murdoch Institute For Research Into Birth Defects Limited ACN 006 566 972 and its controlled entity.

NON-CURRENT LIABILITIES Accounts Payable Provisions TOTAL NON-CURRENT LIABILITIES

This is presented in abridged form: the complete financial report is available on

TOTAL LIABILITIES

3,053,751

2,234,222

1,412,437

1,236,629

NET ASSETS

17,167,448

14,053,704

17,056,189

13,950,506

MEMBERS' FUNDS Accumulated Funds Reserves TOTAL MEMBERS' FUNDS

16,767,448 400,000 17,167,448

14,053,704

16,656,189 400,000 17,056,189

13,950,506

request. telephone +61 3 9345 5045.

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INSTITUTE

Profit and Loss Accounts

Statements of Cash Flows The Company 1997 1996

Consolidated 1997 1996

Operating Profit/(Loss) before Income Tax

2,713,744

5,921,214

2,705,683

5,831,275

Consolidated 1997 1996 Inflows Inflows (Outflows) (Outflows)

The Company 1997 1996 Inflows Inflows (Outflows) (Outflows)

(7,656,003) 3,784,634 6,957,446 1,163,163 20,630 765,311

(5,280,157) 1,465,987 4,025,919 1,543,505 8,778

(4,306,857) 1,374,434 6,957,446 1,038,464 7,659

Patient fees received

(9,009,762) 3,872,592 4,025,919 2,332,730 27,267 1,023,737

NET CASH PROVIDED/(USED) BY OPERATING

2,272,483

5,035,181

1,764,032

5,071,146

Interest received

1,106,159

722,831

1,088,211

705,611

Dividends received

103,120

85,504

103,120

85,504

Proceeds on sale of investments

3,622,833

3,289,742

3,351,540

3,025,468

Payment for investments

(5,892,382)

(7,401,122)

(5,617,294)

(7,129,829)

(566,650)

(192,353)

(511,650)

(142,931)

(1,626,920)

(3,495,398)

(1,586,073)

(3,456,177)

645,563

1,539,783

177,959

1,614,969

2,433,889

894,106

2,412,188

797,219

3,079,452

2,433,889

2,590,147

2,412,188

CASHFLOWS FROM OPERATING ACTIVITIES

Income tax attributable to Operating Loss

Payments to suppliers and employees Government grants received Operating Profit/(Loss) after Income Tax

2,713,744

5,921,214

2,705,683

5,831,275

Accumulated Funds at beginning of the

14,053,704

8,132,490

13,950,506

8,119,231

Donations received Other receipts

financial year

Accumulated Funds at financial year end

14,053,704

16,767,448

16,656,189

13,950,506

Interest received

ACTIVITIES CASH FLOWS FROM INVESTING ACTIVITIES

EXPENDITURE DISTRIBUTION 2%1%

Payment for property, plant and equipment NET CASH PROVIDED BY INVESTING ACTIVITIES ■CUNICMSKWCtS DOWSE INCOME ■ RESEAJKW GRANTS ■ 00NATO4 ■ iNVSSTMCNT income

NET INCREASE IN CASH HELD CASH AT THE BEGINNING OF THE REPORTING

Irevenue receipts u

OPERATING EXPENDITURE

PERIOD

CASH AT THE END OF THE REPORTING PERIOD

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The Murdoch Institute for Research into Birth Defects Limited Royal Children's Hospital Flemington Road PARKVILLE VICTORIA 3052 ACN 006 566 972 Postal Address Post Office Box 1100 PARKVILLE VICTORIA 3052 Telephone: Facsimilie:

(03) 9345 5045 (03) 9348 1391

Produced by Anne Cronin Julie Foletta Voula Pashalidis Michele Winsor

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