NEW YORK • Mr Donald Trump's suggestion that a Fox News journalist had grilled him at the Republican presidential debate because she was menstruating cost him a speaking slot at an influential gathering of conservatives in Atlanta.
It also raised new questions about how much longer Republican Party leaders would have to contend with Mr Trump's disruptive presence.
Continuing his complaints in an interview on CNN last Friday night about Ms Megyn Kelly, one of the moderators of the debate, Mr Trump said: "You could see there was blood coming out of her eyes, blood coming out of her wherever."
Mr Trump had already called out Fox News' moderators for asking "unfair" questions during last Thursday's debate after Ms Kelly brought up his use of derogatory language towards women in Twitter posts. Ms Kelly asked him if this was befitting a man vying to be the United States president.
Mr Trump's remark prompted Mr Erick Erickson, the leader of RedState, the conservative group, to cancel his invitation. "If your standard-bearer has to resort to that... " Mr Erickson told hundreds of conservative activists in a packed Atlanta hotel ballroom last Saturday, "we need a new standard-bearer."
With Mr Trump at centre stage last Thursday in Cleveland, Fox News shattered television viewership records for a primary debate: Nearly 24 million people watched.
But any hopes that Mr Trump, the real estate developer and television celebrity, would try to reinvent himself as a sober-minded statesman, or that he would collapse under scrutiny and tough questions, vaporised in the opening minutes when he refused to rule out running as an independent candidate for president.
Mr Trump denied last Saturday that he had been implying Ms Kelly was menstruating.
"I think only a degenerate would think that I would have meant that," he said in an interview, insisting that he had been referring to Ms Kelly's nose and ears.
But as his latest eruption rippled through Republican circles, the conversation turned to whether the party and his rival presidential contenders should continue to accommodate his candidacy, quietly hoping that this would be the moment he burned out - or whether they should try to run him out on a rail.
If party leaders saw danger in provoking a break-up - and no small advantage to be seized from the ratings bonanza Mr Trump showed himself capable of delivering - there were signs that other influential Republicans would tolerate only so much of Mr Trump's behaviour. Ms Carly Fiorina, former Hewlett-Packard chief executive, who delivered perhaps the most assertive turn in last Thursday's debate among the candidates trailing in the polls, posted on Twitter: "Mr Trump: There. Is. No. Excuse."
Meanwhile, Mr Trump's campaign said on Saturday he had sacked his top political adviser Roger Stone who, instead, said he had resigned because the tenor of the campaign was distracting from Mr Trump's core message.
In an interview last Friday afternoon, before he went on CNN, Mr Trump said he was irritated by the debate moderators' questions about a third-party candidacy, saying he wanted to run as a Republican, but he reiterated his threat to mount one if he is unhappy with his treatment by party leaders.
An independent candidacy would be complicated and costly, he said, but "if you're rich, it's doable". It also appeared unlikely that any network could be persuaded to exclude him. As Mr Trump crowed last Friday in a telephone interview, "I'm a ratings machine."
NEW YORK TIMES, AGENCE FRANCE-PRESSE