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The 2016 WHO Classification of Tumors of the Central Nervous System includes “diffuse midline glioma with histone H3 K27M mutation” as a new diagnostic entity. This study of 33 patients with diffuse midline gliomas found histone H3 K27M mutation was present in 24 patients (72.7%) and absent in 9 (27.3%). The location was the thalamus in 27.3%; the pons in 42.4%; within the vermis/fourth ventricle in 15%; and the spinal cord in 6%. The radiographic features of diffuse midline gliomas with histone H3 K27M mutation were highly variable, ranging from expansile masses without enhancement or necrosis with large areas of surrounding infiltrative growth to peripherally enhancing masses with central necrosis with significant mass effect.