A 16 bit application is any software written for MS-DOS or early versions of Microsoft Windows which originally ran on the 16-bit Intel 8088 and Intel 80286 microprocessors. So generally, it is assumed that 16-bit software is DOS based.
You can run SFC (System File Checker) which scans the drive for altered and missing system files.
Go to Start>Run, type in "SFC /scannow" (without the quotes), click OK. It may ask for your Windows CD, so have it handy.
More details about running SFC are on
this page.