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Original Article

Miltefosine-Induced Apoptotic Cell Death on Leishmania major and L. tropica Strains

Shahram Khademvatan1,2, Mohammad Javad Gharavi3, Fakher Rahim2,4, Jasem Saki1,2
The Korean Journal of Parasitology 2011;49(1):17-23.
Published online: March 18, 2011

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Corresponding author (jasem.saki@gmail.com)
• Received: December 28, 2010   • Revised: January 23, 2011   • Accepted: January 25, 2011

© 2011, Korean Society for Parasitology

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Miltefosine-Induced Apoptotic Cell Death on Leishmania major and L. tropica Strains
Korean J Parasitol. 2011;49(1):17-23.   Published online March 18, 2011
DOI: https://doi.org/10.3347/kjp.2011.49.1.17
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Miltefosine-Induced Apoptotic Cell Death on Leishmania major and L. tropica Strains
Korean J Parasitol. 2011;49(1):17-23.   Published online March 18, 2011
DOI: https://doi.org/10.3347/kjp.2011.49.1.17
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Miltefosine-Induced Apoptotic Cell Death on Leishmania major and L. tropica Strains
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Fig. 1 The viability of L. major and L. tropica promastigotes, in the presence of various concentrations of HePC, assessed by MTT. Each point represents the means of 3 independent tests. Dotted line: L. tropica promastigotes, solid line: L. major.
Fig. 2 Number of promastigotes treated with or without miltefosine at different time points after 48 hr. Dotted line: L. major (MRHO/IR/75/ER) promastigotes and L. tropica (MHOM/IR/02/Mash10) control group, solid line: L. tropica and L. major-treated cells.
Fig. 3 Analysis of morphology in light microscopy (magnification, ×100) and flow cytometry analysis of L. major and L. tropica promastigotes following treatment with 22 µM and 11 µM HePC, respectively, at different time points after miltefosine treatment. (A) L. major 24 hr after treatment. (B) L. major 48 hr after treatment. (C) L. tropica 24 hr after treatment. (D) L. tropica 48 hr after treatment. Lower right region (LR) belongs to apoptotic cells (annexin positive) and upper left region (UL) belongs to necrotic cells (PI positive).
Fig. 4 DNA fragmentation detected with agarose gel electrophoresis of L. major (right) and L. tropica (left). C, Not treated; Lanes 12, 24, 36, and 48 hr incubation periods after treatment with miltefosine. Lane M, size marker.

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Miltefosine-Induced Apoptotic Cell Death on Leishmania major and L. tropica Strains
Species
L. tropica (%)
L. major (%)
Group sub-G0/ G1 (M1) G0/G1 (M2) G2/M (M3) sub-G0/ G1 (M1) G0/G1 (M2) G2/M (M3) 24 hr  Control 3.2 57.7 39.1 4.1 47.3 48.6  Treated 11.5 62.7 25.8 6.5 73.3 20.2 48 hr  Control 3.0 64.9 32.1 3.9 67.0 29.1  Treated 57.1 32.2 10.7 61.2 25.1 13.7
Table 1. The cell cyclea of L. tropica and L. major after treatment with 24 μM and 32 μM miltefosine for 24 hr and 48 hr, respectively

Methods for cell-cycle analysis are described in Materials and Methods.

Table 1.

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