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I get 'Error code 65' when trying to use MTU Assistant.

TweakMASTER Knowledge Base article KB0122

If you are receiving the Error code 65, this is EHOSTUNREACH, no route to host. Typically, this message occurs when an ICMP Host Unreachable message is received from a gateway in response to a packet sent to a destination address. This is sometimes caused by transient network connectivity problems, and is sometimes deliberate on the part of organizations filtering certain kinds of network traffic.

In short, if your ISP filters ICMP, MTU Assistant won't work. To finally decide whether it's TM's fault or not, you should try to ping these hosts with TweakMASTER's ping utility and see if PING works or not.