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I think it does play with how all those tough guy characters were always rejecting women.
Reason is a pursuit, not a conclusion.

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Actually, it was Mae West that originated that line. Just can't remember the movie it was from.

No, it comes from "To Have and Have Not" which was the first pairing of Bogie and Bacall.

But he was in disguise in disguise in disguise

"I never did nothin' to a man that I wouldn't do to a woman!"
I love that line! So gay!
I asked the doctor to take your picture so I can look at you from inside as well.

The implied suggestion that Sam Diamond is a closet homosexual is a spin on and a comic reaction to the macho attitudes displayed by the Philip Marlowe private detective in Raymond Chandler's brilliant series of novels, as well as a play on the iconic, white dinner-jacketed image of Bogie from Casablanca. What makes this character extremely funny is how Peter Falk plays him
straight

• that is, he plays the character as if he's appearing in a serious crime film noir. The muscle magazines give the game away, as well as the sexist remarks he makes about his female sidekick, over-egging his heterosexual persona. In my opinion he delivers the funniest line in the whole movie, when he refutes a remark questioning his sexuality - "
  I never did nothin' to a man that I wouldn't do to a woman!
  " - delivered so deadpan it had me in pain with laughter.

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In the final scene where Sam & assistant are driving away it seems he is really straight and is letting her know that he's available. "All you got to do is whistle. You know how to whistle, don't you?" Which is actually a quote from a Bogart/Bacall movie: "Key Largo" I think. But, yes, his sexuality is a little ambiguous during the movie: the tough guy who won't kiss a "broad."

But he has does have all those muscleman magazines in his office..
Suspects my foot
"Someone has been tampering with Hank's memories."

suspects, always looking for suspects

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It's supposed to be a parody of Bogie's screen image as well- he was such a tough guy, and unquestionably attracted scores of "dames". But if he were 'in the closet'. . .
"Love's not about what you expect to get, only what you expect to give- which is everything."

I don't think Sam is really gay, though.
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My friend, he took his final breath
Now I know the perfect kiss is the kiss of death.

No, I think the final joke about Sam Diamond is he's a closet homosexual, perhaps thats why he overdoes the tough guy bit.
Nobody expects the Spanish Inquisition!!
Michael Palin (Monty Python's Flying Circus)