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BIOSCIENCE REPORTS
ACCEPTED MANUSCRIPT

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ͳ…Šƒ‡Žƒ…–‹˜ƒ–‹‘Ǥ‘ŽŽ‡…–‹˜‡Ž›ǡ–Š‡•‡ˆ‹†‹‰••—‰‰‡•–ƒ†›ƒ‹…”‘Ž‡’Žƒ›‡†
„›–Š‡Ǧ‰Ž›…‘•›Žƒ–‹‘‘ˆŠ—ƒͳǤŠ‡›ƒŽ•‘’”‘˜‹†‡ˆ—”–Š‡”‡˜‹†‡…‡‘ˆ–Š‡
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”‡‰—Žƒ–‹‘‘ˆͳƒ…–‹˜‹–›Ǥ

Cite as Bioscience Reports (2016) DOI: 10.1042/BSR20160149

Copyright 2016 The Author(s).


This is an Accepted Manuscript; not the final Version of Record. You are encouraged to use the final Version of Record that, when
published, will replace this manuscript and be freely available under a Creative Commons licence.
All other rights reserved.
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