According to the British news agency Reuters, the Pentagon said that Lloyd Austin expressed his condolences for the November 13 attack in Istanbul by phone.
They also expressed concern about the escalation of operations in northern Syria and Turkey, including recent airstrikes, the Pentagon said in a statement.
"Some of these (attacks) pose a direct threat to the safety of US personnel working with local partners in Syria to defeat ISIS."
Defense Secretary Lloyd Austin called for a de-escalation and expressed the ministry's strong opposition to Turkey's new military operation in Syria, the statement added.
An explosion in a popular tourist area of Istanbul last November killed six and injured 81 others.
"According to our investigation, the PKK terrorist organization is responsible (for the explosion)," Turkish Interior Minister SĂĽleyman Soylu said in a statement carried by the state-run Anadolu News Agency.
Police officers in Turkey have arrested a Syrian woman who police say has Syrian citizenship.
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