The development comes on sidelines of April 25 meeting of the Joint Commission overseeing implementation of the Joint Comprehensive Plan of Action, in Vienna, where the US delegation raised serious concerns with the Iranian side regarding the cases of US citizens "detained and missing in Iran", said State Department Acting Spokesman Mark Toner.
"The US delegation had called on Iran to immediately release these US citizens so they can be reunited with their families," he said.
Iranian-American businessman Siamak Namazi and his father Baquer Namazi are serving 10-year prison sentences in Iran.
Iran does not recognise dual nationality. Also, the Middle Eastern country rarely allows consular visits to dual nationals in its jails.
"The United States is deeply concerned about reports of their declining health and well-being in detention. The United States urges Iran to immediately release Siamak and Baquer on humanitarian grounds," Toner said in a statement.
"It has also been more than a decade since Robert Levinson disappeared from Iran's Kish Island. Iran is committed to cooperating with the United States in bringing Bob home and we call on Iran to fulfil this commitment.
"The United States remains unwavering in its efforts to return Bob to his family," he said.
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