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Victor Horsley was born in Kensington, London, and educated at Cranbrook School in Kent and at University College London, where he studied medicine under John Burdon Sanderson and G D Thane. In 1880 he was appointed House Surgeon at University College Hospital where he experimented with anaesthetics. Horsley studied at postgraduate level in Berlin in 1881 and in 1882 was appointed Surgical Registrar at University College Hospital. From 1884 to 1890 Horsley was Professor-Superintendent of the Brown Institute, where he did experiments on localization of brain function (with Charles Beevor), on the pituitary gland, on the relation of the larynx to the nervous system (with Felix Semon), and on the thyroid gland, myxoedema and cachexia strumipriva. In 1885 he was promoted to assistant surgeon. In 1886 he took the position of Assistant Professor of Surgery at the National Hospital for Paralysis and Epilepsy, Queen Square, where he performed operations on the brain and spinal cord. In 1886 he was appointed secretary of the Local Government Board Commission on Hydrophobia, and also studied Pasteur's anti-rabies vaccine. In the same year he was elected Fellow of the Royal Society.
From 1887 to 1896 Horsley was Professor of Pathology at University College London. He married Eldred, daughter of Sir Frederick Bramwell, in 1887, and the couple had two sons and one daughter. Horsley was elected President of the Medical Defence Union in 1893 and the British Medical Temperance Association in 1896. In 1897 he was appointed to the Senate of the University of London and elected to the General Medical Council. From 1899 to 1902 he was Professor of Clinical Surgery at University College London. In 1902 he was knighted for his work in medicine. In 1907 he published Alcohol and the Human Body with Dr Mary Sturge. Towards the end of his life he stood as a Liberal candidate in London but later resigned; he was also rejected by Leicester. In 1915 and 1916 he travelled extensively in a medical capacity, performing surgery on the war field. He died at Amara from heatstroke and pyrexia in July 1916.