Ann Woolcock graduated in medicine from the University of Adelaide and pursued postgraduate studies in respiratory medicine with Professor John Read at the University of Sydney. Her MD thesis, awarded in 1967, was on the mechanical behaviour of the lungs in asthma. From 1966 to 1968 she worked with Professor Peter Macklem at McGill University in Canada, then returned to the University of Sydney to continue researching asthma. Her work in asthma and epidemiology showed that asthma was caused by allergens but that there is a genetic component. Her clinical research was a major contribution to better outcomes in asthma, in particular, the demonstration and practical measurement of airway hyperresponsiveness and her subsequent research that examined its contribution to asthma severity and the ways in which treatments were able to reduce it. In 1989 she wrote, with others, the world's first national guidelines for asthma management, the Australian Asthma Management Plan. In 1984, she was appointed to a personal chair of Respiratory Medicine at the University of Sydney. She founded the Institute of Respiratory Medicine in 1985, based at Sydney's Royal Prince Alfred Hospital. After her death, the Institute was renamed the Woolcock Institute of Medical Research in her honour.

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