I prefer to open a new file without creating an audio folder. The documentation states that an audio folder will not be created with a new file by unchecking the box "Use Per-Project Audio Folders" located in Preferences-File-Audio Data. Since installing the Gloucester update, this option no longer appears. Am I missing some vital step or is this a bug?

I noticed "use per project audio folder" check box is moved to the "save as" menu dialog, just do a "save as" and uncheck the box there... (at the bottom of the save as menu)

Thanks for the response; however, this does not prevent the audio folder dialogue from appearing when creating a new file. In previous versions, unchecking the "use per-project audio folder" box in preferences would stop the audio folder request from being displayed. If I'm reading the updated documentation correctly, it appears that when creating a new project you will always be prompted to name the file and create an audio folder. By not entering a project name, an audio folder is not created.
(I still prefer the old method of disabling per-project folders altogether, but I guess the bakers have their reasons.)

Use the new Start Screen and read the eZine documentation. You can create new projects that default to not creating a folder.

Thanks! That answers my question and resolves the issue.