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Morphological characteristics of apoptosis-like death including DNA fragmentation, increased vacuolization, nuclear condensation and cell rounding were observed for H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub>-exposed trophozoites with preservation of membrane integrity. Biochemical alteration in ion fluxes is also a key feature in PCD, and H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub>-exposed trophozoites showed overproduction of reactive oxygen species, increased cytosolic Ca<jats:sup>2+<\/jats:sup>and decreased intracellular pH. Phosphatidylserine was also found to be expressed in the outer leaflet of the plasma membrane of the H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub>-treated trophozoites. Pretreatment with the cysteine protease inhibitor E-64d, the extracellular and intracellular Ca<jats:sup>2+<\/jats:sup>chelators EGTA and BAPTA\/AM, and the Ca<jats:sup>2+<\/jats:sup>influx inhibitor verapamil prior to H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub>exposure abolished DNA fragmentation. The oxidatively stressed trophozoites also showed an increased calpain activity, indicating involvement of Ca<jats:sup>2+<\/jats:sup>-dependent calpain-like cysteine proteases in PCD of<jats:italic>E. histolytica<\/jats:italic>. A homogeneous caspase assay showed no significant caspase activity, and administration of caspase 1 inhibitor also did not prevent the death phenotype for the oxidatively stressed trophozoites, indicating a caspase-independent apoptosis-like death. Our observations clearly demonstrate that there is a distinct calpain-dependent but caspase-independent pathway for apoptosis-like death in oxidatively stressed<jats:italic>E. histolytica<\/jats:italic>trophozoites.<\/jats:p>","DOI":"10.1099\/mic.0.034066-0","type":"journal-article","created":{"date-parts":[[2010,3,19]],"date-time":"2010-03-19T00:34:49Z","timestamp":1268958889000},"page":"1926-1941","source":"Crossref","is-referenced-by-count":22,"title":["Hydrogen peroxide induces apoptosis-like death in Entamoeba histolytica trophozoites"],"prefix":"10.1099","volume":"156","author":[{"given":"Nilay","family":"Nandi","sequence":"first","affiliation":[{"name":"Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna-800007, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abhik","family":"Sen","sequence":"additional","affiliation":[{"name":"Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna-800007, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rajdeep","family":"Banerjee","sequence":"additional","affiliation":[{"name":"Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna-800007, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sudeep","family":"Kumar","sequence":"additional","affiliation":[{"name":"Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna-800007, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vikash","family":"Kumar","sequence":"additional","affiliation":[{"name":"Division of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna-800007, Bihar, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amar Nath","family":"Ghosh","sequence":"additional","affiliation":[{"name":"Division of Electron Microscopy, National Institute of Cholera and Enteric Diseases, P-33, C.I.T. 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