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</o:shapelayout></xml><![endif]--></head><body lang=EN-US link=blue vlink=purple><div class=WordSection1><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Sunil:<o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'><o:p> </o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Axenic amastigotes (Ax) are much more than “stressed-out promastigotes”. We (and others) have shown they are very similar (although not identical) to macrophage-derived amastigotes (Am) in terms of global mRNA and protein expression levels. While it would obviously be better to work with Am, it is often technically difficult due to low numbers and contamination with macrophage RNA and protein. In addition, it is my contention that the time taken to isolate Am from the macrophages makes them “stressed-out amastigotes”, which are less physiological relevant than Ax. We need to approach this discussion from a position of knowledge and data (i.e. understanding exactly which genes are expressed differently in Am and Ax) than belief. Hopefully, we will be able to publish the results of our RNA-seq studies comparing promastigotes, axenic amastigotes and macrophage-derived amastigotes (without purification) in the not too distant future.<o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'><o:p> </o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Peter <o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'><o:p> </o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>Peter J. Myler, Ph.D.</span><span style='color:navy'><o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>Professor</span><span style='color:navy'><br></span><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>Seattle BioMed<br>307 Westlake Ave N., Suite 500<br>Seattle, WA 98109-5219<br>USA</span><span style='color:navy'> <br></span><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>(206) 256-7332 (office)</span><span style='color:navy'><br></span><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>(206) 256-7229 (FAX)</span><span style='color:navy'> <br></span><span style='color:#1F497D'><a href="mailto:peter.myler@sbri.org">peter.myler@sbri.org</a><o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span style='color:navy'>Affiliate Professor <br>Departments of Global Health &<br>Biomedical Informatics and Medical Education <br>University of Washington <br>Seattle, WA 98195 <br>Box 357238 <br></span><span style='color:#1F497D'><a href="mailto:mylerpj@u.washington.edu">mylerpj@u.washington.edu</a></span><span style='color:navy'> <o:p></o:p></span></p></div><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'><o:p> </o:p></span></p><div><div style='border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0in 0in 0in'><p class=MsoNormal><b><span style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'>From:</span></b><span style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'> leish-l-bounces@lineu.icb.usp.br [mailto:leish-l-bounces@lineu.icb.usp.br] <b>On Behalf Of </b>Sunil Arora<br><b>Sent:</b> Monday, March 11, 2013 8:42 AM<br><b>To:</b> Clos Joachim; Leish-L email post<br><b>Subject:</b> Re: [Leish-l] Fwd: hot spots<o:p></o:p></span></p></div></div><p class=MsoNormal><o:p> </o:p></p><div><div><p class=MsoNormal style='background:white'><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'>I agree...axenic amastigotes are more like stressed out promastigotes... I always prefer working with something inside the macrophages as a model for testing drugs or immunomodulators which I believe are closer to actual intracellular amastigotes in the infected host<o:p></o:p></span></p></div><div><p class=MsoNormal><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'><o:p> </o:p></span></p></div><div><p class=MsoNormal><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'>best<o:p></o:p></span></p></div><div><p class=MsoNormal><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'>sunil<o:p></o:p></span></p></div><div><p class=MsoNormal style='background:white'><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'> <o:p></o:p></span></p></div><div><p class=MsoNormal style='background:white'><i><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:#0000BF'>---------------------------------------------------------------</span></i><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'><o:p></o:p></span></p></div><div><p class=MsoNormal style='background:white'><i><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:#0000BF'>Dr Sunil K.Arora<br>Professor <br>In Charge- HIV Diagnostic and Disease Monitoring Laboratory &<br>NACO State Reference Laboratory,</span></i><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'><o:p></o:p></span></p></div><div><p class=MsoNormal style='background:white'><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'> <o:p></o:p></span></p></div><div><p class=MsoNormal style='background:white'><i><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:#0000BF'>Councillor- Federation of Immunological Societies of Asia-oceania (FIMSA)</span></i><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'><o:p></o:p></span></p></div><div><p class=MsoNormal><i><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:#0000BF'>Vice President- Indian Immunology Society (IIS)</span></i><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:#0000BF'><o:p></o:p></span></p></div><div><p class=MsoNormal><i><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:#0000BF'>Secretary (Research)- The Cytometry Society of India (TCS)</span></i><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:#0000BF'><o:p></o:p></span></p></div><div><p class=MsoNormal><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:#0000BF'> <o:p></o:p></span></p></div><div><p class=MsoNormal><i><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:#0000BF'>Department of Immunopathology<br>PGIMER, Chandigarh-160 012<br>Ph.: 0091-172-2755192 (Off)<br>FAX: 0091-172-2744401, 2745078<br>Ph:+91-172-4666087 (Res); 9872866087 (cell)<br>email: <a href="mailto:skarora_in@yahoo.com" target="_blank">skarora_in@yahoo.com</a> ; <a href="mailto:skarorain@gmail.com" target="_blank">skarorain@gmail.com</a> </span></i><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:#0000BF'><o:p></o:p></span></p></div><div><p class=MsoNormal style='background:white'><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'><o:p> </o:p></span></p></div><div><div><div><div class=MsoNormal align=center style='text-align:center;background:white'><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'><hr size=1 width="100%" align=center></span></div><p class=MsoNormal style='background:white'><b><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>From:</span></b><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'> Clos Joachim <clos@bni-hamburg.de><br><b>To:</b> Leish-L email post <Leish-l@lineu.icb.usp.br> <br><b>Sent:</b> Monday, 11 March 2013 8:16 PM<br><b>Subject:</b> [Leish-l] Fwd: hot spots</span><span style='color:black'><o:p></o:p></span></p></div><p class=MsoNormal style='background:white'><span style='color:black'><o:p> </o:p></span></p><div id=yiv1658093374><div><div><blockquote style='margin-top:5.0pt;margin-bottom:5.0pt'><div><p class=MsoNormal style='background:white'><span style='font-size:10.0pt;font-family:"Baghdad","serif";color:#010101'>Are axenic amastigotes a valid model (and for what?), the first developmental step towards real amastigotes, or just stressed-out promastigotes? Many of us use axenics and swear by them, others are skeptical, others will accept what's inside a macrophage, still others only want amastigotes isolated from something with a fur. I believe we should come to some sort of common language and interpretation. I am somewhere in the middle myself, preferring to do things in macrophages whenever possible.</span><span style='color:black'><o:p></o:p></span></p><div><p class=MsoNormal style='background:white'><span style='font-size:10.0pt;font-family:"Baghdad","serif";color:#010101'>Cheers!</span><span style='color:black'><o:p></o:p></span></p></div></div></blockquote></div><div><p class=MsoNormal style='background:white'><span style='color:black'><br><br><o:p></o:p></span></p><div><div><p class=MsoNormal style='background:white'><span style='color:black'><o:p> </o:p></span></p></div><div><p class=MsoNormal style='background:white'><span style='color:black'><o:p> </o:p></span></p><div><div><p class=MsoNormal style='background:white'><span style='color:black'>Am 07.02.2013 um 20:38 schrieb Carlos Costa:<o:p></o:p></span></p></div><p class=MsoNormal style='background:white'><span style='color:black'><br><br><o:p></o:p></span></p><div><p class=MsoNormal style='background:white'><span style='font-family:"Verdana","sans-serif";color:black'>What are the most polemic issues in leishmaniasis?<br clear=all></span><span style='color:black'><o:p></o:p></span></p><div><p class=MsoNormal style='background:white'><span style='color:black'><o:p> </o:p></span></p></div><p class=MsoNormal style='background:white'><span style='color:black'>-- <o:p></o:p></span></p><p class=MsoNormal style='background:white'><span style='font-family:"Verdana","sans-serif";color:black'>Carlos H. N. Costa, MD, DSc.</span><span style='color:black'><o:p></o:p></span></p><p class=MsoNormal style='background:white'><span style='font-family:"Verdana","sans-serif";color:black'>Sociedade Brasileira de Medicina Tropical </span><span style='color:black'><o:p></o:p></span></p><p class=MsoNormal style='background:white'><span style='font-family:"Verdana","sans-serif";color:black'>(Brazilian Society of Tropical Medicine)</span><span style='color:black'><o:p></o:p></span></p><p class=MsoNormal style='background:white'><span style='font-family:"Verdana","sans-serif";color:black'>President</span><span style='color:black'><o:p></o:p></span></p><p class=MsoNormal style='background:white'><span style='color:black'><o:p> </o:p></span></p><p class=MsoNormal style='background:white'><span style='font-family:"Verdana","sans-serif";color:black'>Universidade Federal do Piauí</span><span style='color:black'><o:p></o:p></span></p><div><p class=MsoNormal style='background:white'><span style='font-family:"Verdana","sans-serif";color:black'>Instituto de Doenças Tropicais Natan Portella</span><span style='color:black'><o:p></o:p></span></p><p class=MsoNormal style='background:white'><span style='font-family:"Verdana","sans-serif";color:black'>Rua Artur de Vasconcelos 151-Sul</span><span style='color:black'><o:p></o:p></span></p><div><p class=MsoNormal style='background:white'><span style='font-family:"Verdana","sans-serif";color:black'>64001-450 Teresina-PI</span><span style='color:black'><o:p></o:p></span></p><p class=MsoNormal style='background:white'><span style='font-family:"Verdana","sans-serif";color:black'>Brazil<o:p></o:p></span></p><p class=MsoNormal style='background:white'><span style='font-family:"Verdana","sans-serif";color:black'>Telephones: +55 86 3222-4377 (W), <o:p></o:p></span></p><p class=MsoNormal align=center style='text-align:center;background:white'><span style='font-family:"Verdana","sans-serif";color:black'>+55 86 3221-3062 (W), <o:p></o:p></span></p><p class=MsoNormal align=center style='text-align:center;background:white'><span style='font-family:"Verdana","sans-serif";color:black'>+55 86 3237-1075 (R). <o:p></o:p></span></p><p class=MsoNormal style='background:white'><span style='font-family:"Verdana","sans-serif";color:black'><o:p> </o:p></span></p><p class=MsoNormal style='background:white'><span style='font-family:"Verdana","sans-serif";color:black'><o:p> </o:p></span></p><p class=MsoNormal style='background:white'><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'>Aviso: As informações contidas nesta mensagem são CONFIDENCIAIS, protegidas pelo sigilo legal, por direitos autorais e destinadas exclusivamente à pessoa ou organização para a qual a mensagem foi destinada.<o:p></o:p></span></p><p class=MsoNormal style='background:white'><span style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'>Warning: This message is meant only for the intended recipient of the transmission. 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If you are not the correct recipient, please notify us immediately by return e-mail and delete this message from your system.<o:p></o:p></span></p></div></div></div><p class=MsoNormal style='background:white'><span style='color:black'>_______________________________________________<br>Leish-l mailing list<br><a href="mailto:Leish-l@lineu.icb.usp.br" target="_blank">Leish-l@lineu.icb.usp.br</a><br><a href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l">http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l</a><o:p></o:p></span></p></div><p class=MsoNormal style='background:white'><span style='color:black'><o:p> </o:p></span></p></div><p class=MsoNormal style='margin-bottom:12.0pt;background:white'><span style='color:black'><o:p> </o:p></span></p><div><div><p class=MsoNormal style='background:white'><span class=yiv1658093374apple-style-span><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif";color:black'>Joachim Clos, Dr. rer. nat.</span></span><span style='font-size:9.0pt;font-family:"Arial","sans-serif";color:black'><o:p></o:p></span></p></div><div><p class=MsoNormal style='background:white'><span class=yiv1658093374apple-style-span><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif";color:black'>Senior Staff Scientist</span></span><span style='font-size:9.0pt;font-family:"Arial","sans-serif";color:black'><o:p></o:p></span></p></div><div><p class=MsoNormal style='background:white'><span class=yiv1658093374apple-style-span><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif";color:black'>Group Leader</span></span><span style='font-size:9.0pt;font-family:"Arial","sans-serif";color:black'><o:p></o:p></span></p></div><div><p class=MsoNormal style='background:white'><span class=yiv1658093374apple-style-span><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif";color:black'>Bernhard Nocht Institute for Tropical Medicine</span></span><span style='font-size:9.0pt;font-family:"Arial","sans-serif";color:black'><o:p></o:p></span></p></div><div><p class=MsoNormal style='background:white'><span class=yiv1658093374apple-style-span><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif";color:black'>Bernhard Nocht St. 74, D-20359 Hamburg, FRG</span></span><span style='font-size:9.0pt;font-family:"Arial","sans-serif";color:black'><o:p></o:p></span></p></div><div><p class=MsoNormal style='background:white'><span class=yiv1658093374apple-style-span><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif";color:black'>Phone +49 40 42818-481; Fax -512</span></span><span style='font-size:9.0pt;font-family:"Arial","sans-serif";color:black'><o:p></o:p></span></p></div><div><p class=MsoNormal style='background:white'><span class=yiv1658093374apple-style-span><span style='font-size:9.0pt;font-family:"Helvetica","sans-serif";color:black'>Skype: joachim.clos</span></span><span style='font-size:9.0pt;font-family:"Arial","sans-serif";color:black'><o:p></o:p></span></p></div><p class=MsoNormal style='background:white'><span style='font-size:9.0pt;font-family:"Arial","sans-serif";color:black'><br><br></span><span style='color:black'><o:p></o:p></span></p></div><p class=MsoNormal style='background:white'><span style='color:black'><o:p> </o:p></span></p></div></div><p class=MsoNormal style='background:white'><span style='color:black'><br><br><br>--<br><i>This email was sent by icb.usp.br </i> <o:p></o:p></span></p></div></div><p class=MsoNormal style='margin-bottom:12.0pt;background:white'><span style='color:black'><br>_______________________________________________<br>Leish-l mailing list<br><a href="mailto:Leish-l@lineu.icb.usp.br">Leish-l@lineu.icb.usp.br</a><br><a href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l" target="_blank">http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l</a><br><br>--<br>This email was sent by icb.usp.br<br><br><o:p></o:p></span></p></div></div></div><p class=MsoNormal><br><br>--<br><i>This email was sent by icb.usp.br </i> <o:p></o:p></p></div></body></html>