<html><body><div style="color:#000; background-color:#fff; font-family:verdana, helvetica, sans-serif;font-size:10pt"><div style="RIGHT: auto"><SPAN style="RIGHT: auto">Dear KP</SPAN></div>
<div style="RIGHT: auto"><SPAN style="RIGHT: auto">of course, myself and Dr Sharma have plans to continue our collaborative program further. Dr sharma may have to show the green signal as soon as we get the funding in place..</SPAN></div>
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<div style="RIGHT: auto"><SPAN style="RIGHT: auto">anyways... very nice to have your exclusive comments and suggestions</SPAN></div>
<div style="RIGHT: auto"><SPAN style="RIGHT: auto">regards</SPAN></div>
<div style="RIGHT: auto"><SPAN style="RIGHT: auto">sunil</SPAN></div>
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<DIV style="BORDER-BOTTOM: #ccc 1px solid; BORDER-LEFT: #ccc 1px solid; PADDING-BOTTOM: 0px; LINE-HEIGHT: 0; MARGIN: 5px 0px; PADDING-LEFT: 0px; PADDING-RIGHT: 0px; FONT-SIZE: 0px; BORDER-TOP: #ccc 1px solid; BORDER-RIGHT: #ccc 1px solid; PADDING-TOP: 0px" class=hr contentEditable=false readonly="true"></DIV><B><SPAN style="FONT-WEIGHT: bold">From:</SPAN></B> "Chang, Kwang-Poo" <KwangPoo.Chang@rosalindfranklin.edu><BR><B><SPAN style="FONT-WEIGHT: bold">To:</SPAN></B> Dr.Sunil Arora <skarora_in@yahoo.com>; mmukhtar@tropmedicine.org; Sharman <nandlals@hotmail.com>; Petr Volf <volf@cesnet.cz>; Hiro Goto <hgoto@usp.br>; elfadil abass <elfadil_abass@yahoo.com>; vishwamohan_katoch@yahoo.co.in; lalit Kant ICMR <lalitkant@icmr.org.in><BR><B><SPAN style="FONT-WEIGHT: bold">Cc:</SPAN></B> leish-l@lineu.icb.usp.br; "Chang, Kwang-Poo" <KwangPoo.Chang@rosalindfranklin.edu><BR><B><SPAN style="FONT-WEIGHT:
bold">Sent:</SPAN></B> Friday, June 3, 2011 11:27 PM<BR><B><SPAN style="FONT-WEIGHT: bold">Subject:</SPAN></B> RE: [Leish-l] inquiry<BR></FONT><BR>
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<div class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: navy; FONT-SIZE: 10pt">Dear Sunil,</SPAN></FONT></div>
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<div class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: navy; FONT-SIZE: 10pt">Glad to hear from you ! It is good for you (and anyone else) to chip in with different views/valuable information on the subject under discussion. I wish to say the following:</SPAN></FONT></div>
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<LI style="COLOR: navy" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; FONT-SIZE: 10pt">It is important for you and Dr. Sharma to work together, which is very much in line with the spirit of WorldLeish.</SPAN></FONT></LI></OL>
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<LI style="COLOR: navy" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; FONT-SIZE: 10pt">Leishmania from some CL patients along the Sutluj river valley belong to the L donovani/infantum species complex. As you pointed out, they are refractory to in vitro cultivation and thus very different from those in Bihar . Splenic aspirates from Shyam Sundar’s VL patients produced exuberant growth as promasigotes in 3N, as I recalled from my visit to his clinic in the 1980’s (although none of these promastigotes produces kala-azar in hamsters). This is well-known (correct me if I should be wrong ?). The genetic sites of these and other Bihar isolates we examined are different from L infantum strains studied, including those from China (Xinjiang, Beijing , Shandong , Sichuan , Gansu ), Pakistan , Iran , Turkey and elsewhere (see Waki et al. 2007 Eukaryot. Cell 6, 198). Interestingly, the Bihar isolates is identical
to one Sri Lanka CL isolate (DNA kindly provided by Y Matsumoto, U Tokyo) in nagt sequence, consistent with published report. Most interestingly, Shyam Sundar, JC Dujardin and their colleagues recently reported the isolation of Leptomonas from ~7% (according to Shyam) of his Bihar kala-alar samples. This, coupled with the difficulty of in vitro cultures from clinical samples at least in some cases, underscores the need to directly examine Leishmania in the original sample sources.</SPAN></FONT></LI></OL>
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<LI style="COLOR: navy" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; FONT-SIZE: 10pt">I have followed the convention of naming the causative agents of Indian and Sudanese kala-azar/PKDL as L donovani. The gene sequences differ between the two (we examined) as well as from all infantum isolates (including some previously typed by others as such). However, this L donovani/infantum species complex is the most sequence-conserved group, consisting of only 5 nagt genetic variants with limited base substitutions (see Fig. 6[D-4] in Waki et al.). The sequence divergence can be larger among the infantum variants than between infantum vs donovani. Analyses of their whole genomic sequencing may shed light on their true evolutionary relationships in genetic terms. </SPAN></FONT></LI></OL>
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<LI style="COLOR: navy" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; FONT-SIZE: 10pt">What is the best genetic site for typing Leishmania isolates ? This is the question I have been asked repeatedly. My answer to that is try to analyze sequence divergence in as many sites as possible, preferably the entire genomes. Analyses of most sites can be informative down to the species complex level. Interpretation beyond that (to the subspecies complex level, for example) is often difficult (and thus must remain hypothetical) without a complete database of all existing Leishmania variants. The database is incomplete because we are unable to grow parasites from many places and there are too many endemic sites which have not been explored. </SPAN></FONT></LI></OL>
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<LI style="COLOR: navy" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; FONT-SIZE: 10pt">With regard to the ease of isolating L infantum for continuous cultivation from clinical, sand fly and reservoir samples, different investigators differ in their experience. Dr. Bastien said he has no problem. Carlos Costa also told me that he has no difficulty of growing L chagasi from his patients in Teresina . He does have a sizable collection of hundreds (or thousands ?) of isolates, although I was not given the actual rate of successful isolation.</SPAN></FONT></LI></OL>
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<div class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: navy; FONT-SIZE: 10pt">KP</SPAN></FONT></div>
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<div class=yiv639626477MsoNormal><B><FONT size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; FONT-SIZE: 10pt; FONT-WEIGHT: bold">From:</SPAN></FONT></B><FONT size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; FONT-SIZE: 10pt"> Dr.Sunil Arora [mailto:skarora_in@yahoo.com] <BR><B><SPAN style="FONT-WEIGHT: bold">Sent:</SPAN></B> Thursday, June 02, 2011 11:55 PM<BR><B><SPAN style="FONT-WEIGHT: bold">To:</SPAN></B> Chang, Kwang-Poo; mmukhtar@tropmedicine.org; Sharman; Petr Volf; Hiro Goto; elfadil abass; vishwamohan_katoch@yahoo.co.in; lalit Kant ICMR<BR><B><SPAN style="FONT-WEIGHT: bold">Cc:</SPAN></B> leish-l@lineu.icb.usp.br<BR><B><SPAN style="FONT-WEIGHT: bold">Subject:</SPAN></B> Re: [Leish-l] inquiry</SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Verdana><SPAN style="FONT-FAMILY: Verdana; COLOR: black; FONT-SIZE: 10pt">Dear KP and all</SPAN></FONT></div></DIV><SPAN>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Verdana><SPAN style="FONT-FAMILY: Verdana; COLOR: black; FONT-SIZE: 10pt">I have been reading the exchange of emails regarding characterisation of leishmania isolates from Himachal in India by Dr Sharma and KP. Actually I have also been associated with that though very briefly and had a chance to try-n-culture some of isolates sent by Dr Sharma. The cultures became positive but we failed to establish good passagable cultures from any of aspirates/biopsy specimen sent by Dr Sharma, although my lab has been able to establish some good number of primaray cultures from spleen/BM aspirates of Ld strains in Chandigarh, although almost all the source patients belonged to Bihar originally for these. So there is definetely some thing very different about the leishmania strains that are circulating in that geographical area of India . we also had a
chance to use a specific PCR (that was developed in my lab and can distinguish between an Ld and any cutaneous strain). we found about 40-50% of isolates got Ld specific amplification while others didnot show Ld genome. So I do believe, there is a mix of Ld and some cutaneous strain in that area which KP is also refering too. It needs to be investigated further... I know Dr Sharma is on to such effort soon...</SPAN></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Verdana><SPAN style="FONT-FAMILY: Verdana; COLOR: black; FONT-SIZE: 10pt">regards</SPAN></SPAN></FONT></div></DIV><SPAN>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Verdana><SPAN style="FONT-FAMILY: Verdana; COLOR: black; FONT-SIZE: 10pt">sunil</SPAN></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Verdana><SPAN style="FONT-FAMILY: Verdana; COLOR: black; FONT-SIZE: 10pt"> </SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><I><I><FONT color=#00007f size=2 face=Verdana><SPAN style="FONT-FAMILY: Verdana; COLOR: #00007f; FONT-SIZE: 10pt" class=ms__id8447>Dr Sunil K.Arora</SPAN></FONT></I></I><I><FONT color=#00007f size=2 face=Verdana><SPAN style="FONT-STYLE: italic; FONT-FAMILY: Verdana; COLOR: #00007f; FONT-SIZE: 10pt"><BR><I><I><FONT face=Verdana><SPAN style="FONT-FAMILY: Verdana" class=ms__id8451>Professor </SPAN></FONT></I></I><BR><I><I><FONT face=Verdana><SPAN style="FONT-FAMILY: Verdana" class=ms__id8453>In Charge HIV diagnostic and Disease Monitoring Laboratory</SPAN></FONT></I></I><BR><I><I><FONT face=Verdana><SPAN style="FONT-FAMILY: Verdana" class=ms__id8455>Department of Immunopathology</SPAN></FONT></I></I><BR><I><I><FONT face=Verdana><SPAN style="FONT-FAMILY: Verdana" class=ms__id8457>PGIMER, Chandigarh-160 012</SPAN></FONT></I></I><BR><I><I><FONT face=Verdana><SPAN style="FONT-FAMILY: Verdana"
class=ms__id8459>Ph.: 0091-172-2755192(Off)</SPAN></FONT></I></I><BR><I><I><FONT face=Verdana><SPAN style="FONT-FAMILY: Verdana" class=ms__id8461>FAX: 0091-172-2744401, 2745078</SPAN></FONT></I></I><BR><I><I><FONT face=Verdana><SPAN style="FONT-FAMILY: Verdana" class=ms__id8463>Ph:+91-172-2732863 (Res); 9872866087 (cell)</SPAN></FONT></I></I><BR><I><I><FONT face=Verdana><SPAN style="FONT-FAMILY: Verdana" class=ms__id8465>email: skarora_in@yahoo.com; skarorain@gmail.com</SPAN></FONT></I></I></SPAN></FONT></I><FONT color=black size=2 face=Verdana><SPAN style="FONT-FAMILY: Verdana; COLOR: black; FONT-SIZE: 10pt"></SPAN></FONT></div></DIV>
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<div style="MARGIN-BOTTOM: 12pt; BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">From:</SPAN></FONT></B><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt"> "Chang, Kwang-Poo" <KwangPoo.Chang@rosalindfranklin.edu><BR><B><SPAN style="FONT-WEIGHT: bold">To:</SPAN></B> mmukhtar@tropmedicine.org; Sharman <nandlals@hotmail.com>; Petr Volf <volf@cesnet.cz>; Hiro Goto <hgoto@usp.br>; elfadil abass <elfadil_abass@yahoo.com>; vishwamohan_katoch@yahoo.co.in; lalit Kant ICMR <lalitkant@icmr.org.in><BR><B><SPAN style="FONT-WEIGHT: bold">Cc:</SPAN></B> leish-l@lineu.icb.usp.br<BR><B><SPAN style="FONT-WEIGHT: bold">Sent:</SPAN></B> Friday, June 3, 2011 12:03 AM<BR><B><SPAN style="FONT-WEIGHT: bold">Subject:</SPAN></B> Re: [Leish-l] inquiry</SPAN></FONT><FONT
color=black><SPAN style="COLOR: black"></SPAN></FONT></div>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: navy; FONT-SIZE: 10pt">Dear Maowia,</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: navy; FONT-SIZE: 10pt">Thanks again for the info.</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: navy; FONT-SIZE: 10pt">Are there any record available for the number of successful isolations versus the total number attempted for a given endemic site ?</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: navy; FONT-SIZE: 10pt">KP</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: navy; FONT-SIZE: 10pt"> </SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">From:</SPAN></FONT></B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt"> Maowia Mukhtar [mailto: mmukhtar@tropmedicine.org ] <BR><B><SPAN style="FONT-WEIGHT: bold">Sent:</SPAN></B> Wednesday, June 01, 2011 10:41 PM<BR><B><SPAN style="FONT-WEIGHT: bold">To:</SPAN></B> Chang, Kwang-Poo; 'Sharman'; 'Petr Volf'; 'Hiro Goto'; 'elfadil abass'; vishwamohan_katoch@yahoo.co.in; 'lalit Kant ICMR'<BR><B><SPAN style="FONT-WEIGHT: bold">Cc:</SPAN></B> leish-l@lineu.icb.usp.br<BR><B><SPAN style="FONT-WEIGHT: bold">Subject:</SPAN></B> RE: [Leish-l] inquiry</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=3 face="Times New Roman"><SPAN style="COLOR: black; FONT-SIZE: 12pt"> </SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt">Dear KP,</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt">Thanks for your comments and valuable notes. We have a high success rate inc culturing Leishmania from clinical aspirates. We have even succeeded in culturing Leishmania directly from PKDL lesions without using hamsters although the parasite is known to be scarce in these lesions. We have noticed that the rate of success of cultures varies between endemic regions in Sudan as well as between the clinical forms. Sudanese Leishmania donovani is known to be different from the Indian donovani even in their response to antileishmania drugs and in their reaction using different diagnostic serology tests. </SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt">Best regards</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt">Maowia</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt; FONT-WEIGHT: bold">Professor Maowia M. Mukhtar</SPAN></FONT></B><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt; FONT-WEIGHT: bold">Institute</SPAN></FONT></B><B><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt; FONT-WEIGHT: bold"> of Endemic</SPAN></FONT></B><B><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt; FONT-WEIGHT: bold"> Diseases</SPAN></FONT></B><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt; FONT-WEIGHT: bold">University</SPAN></FONT></B><B><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt; FONT-WEIGHT: bold"> of Khartoum</SPAN></FONT></B><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt; FONT-WEIGHT: bold">P.O. Box 11463, khartoum, Sudan</SPAN></FONT></B><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt; FONT-WEIGHT: bold">Mobile</SPAN></FONT></B><B><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt; FONT-WEIGHT: bold">; +249912234268</SPAN></FONT></B><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">From:</SPAN></FONT></B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt"> Chang, Kwang-Poo [mailto:KwangPoo.Chang@rosalindfranklin.edu] <BR><B><SPAN style="FONT-WEIGHT: bold">Sent:</SPAN></B> 02 June 2011 01:42<BR><B><SPAN style="FONT-WEIGHT: bold">To:</SPAN></B> mmukhtar; Sharman; Petr Volf; Hiro Goto; elfadil abass; vishwamohan_katoch@yahoo.co.in; lalit Kant ICMR<BR><B><SPAN style="FONT-WEIGHT: bold">Cc:</SPAN></B> leish-l@lineu.icb.usp.br; Chang, Kwang-Poo<BR><B><SPAN style="FONT-WEIGHT: bold">Subject:</SPAN></B> RE: [Leish-l] inquiry</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: navy; FONT-SIZE: 10pt">Dear Maowia,</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: navy; FONT-SIZE: 10pt">Gald to hear from you with your work-in-progress. May I ask you some questions and share with you some info from my very limited experience with Sudanese L donovani ? Please correct my statements, if inccorrect.</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<LI style="BACKGROUND: white; COLOR: navy" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; FONT-SIZE: 10pt">“difficult to maintain” means that the promastigotes emerge after incubation of lesion aspirates/punctures in appropriate media and replicate during successive passages, but are low in cell density and easily lost during repetitive passages for continuous cultivation ?</SPAN></FONT></LI></OL>
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<LI style="BACKGROUND: white; COLOR: navy" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; FONT-SIZE: 10pt">“difficult to grow” means that the promastigotes never emerge or emerge after incubation of lesion aspirates/punctures in appropriate media, but subsequently fail to replicate or replicate to a limited extent on subculture ?</SPAN></FONT></LI></OL>
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<LI style="BACKGROUND: white; COLOR: navy" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; FONT-SIZE: 10pt">What is the rate of success in your attempts to isolate parasites in culture from clinical samples in each endemic site over the years ? </SPAN></FONT></LI></OL>
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<LI style="BACKGROUND: white; COLOR: navy" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; FONT-SIZE: 10pt">The Sudanese strain used by many in the USA was probably brought over decades ago by Donald Hyneman and established in Syrian Golden hamsters by Leslie Stauber and his students. In the 70’s, I had the good fortune of working with Dennis Dwyer on his S1 or S2 strains of this origin. As widely reported in the literature, this is an excellent model of visceral leishmaniasis, producing typical clinical outcome consistently, including invariable death and splenic LD loads of up to 10<SUP>10</SUP> per heavily infected spleen. I recall I had difficult time to differentiate these splenic amastigotes into promastigotes. I did see some motile promastigotes. While a small number remains viable, many died and appeared as ghosts during the subsequent incubation. This has led me to suspect selection of cultivable
clones. In some, but not all of these cultures, promastigotes eventually became ‘adapted’ to the culture conditions and grew, producing consistently up to ~40 X 10<SUP>6</SUP> cells/ml in 4-5 days in successive passages (starting from ~10<SUP>6</SUP>/ml = ~ 4-5 generation or replication cycles). The stationary phase promastigotes of these cultures no longer produce good infection of hamsters, irrespective of the passage numbers (Isolation of ‘metacyclics” to produce good infection has since been reported in the literature, if I remember correctly). These promastigotes are more similar to the Mediterranean L infantum than to the Indian L donovani of the Bihar region in the sequence of their single-copy protein-coding genes, as probably already shown by many based on sequencing other genetic sites. </SPAN></FONT></LI></OL>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=navy size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: navy; FONT-SIZE: 10pt"> </SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">From:</SPAN></FONT></B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt"> Maowia Mukhtar [mailto: mmukhtar@tropmedicine.org ] <BR><B><SPAN style="FONT-WEIGHT: bold">Sent:</SPAN></B> Tuesday, May 31, 2011 2:30 PM<BR><B><SPAN style="FONT-WEIGHT: bold">To:</SPAN></B> 'Sharman'; 'Petr Volf'; Chang, Kwang-Poo; 'Hiro Goto'; 'elfadil abass'; vishwamohan_katoch@yahoo.co.in; 'lalit Kant ICMR'<BR><B><SPAN style="FONT-WEIGHT: bold">Cc:</SPAN></B> leish-l@lineu.icb.usp.br<BR><B><SPAN style="FONT-WEIGHT: bold">Subject:</SPAN></B> RE: [Leish-l] inquiry</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=3 face="Times New Roman"><SPAN style="COLOR: black; FONT-SIZE: 12pt"> </SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt">Dear Colleagues,</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt">We have very interesting experience with Sudanese L.donovani. This group of parasite although are genetically homogenous they still cause diverse clinical forms in Sudan ranging from VL, PKDL, ML to CL. We have isolated L. donovani MON82 from classic cutaneous ulcers and typed them using both isoenzyme and molecular typing techniques. Please check our manuscript (Elamin et al. 2008. </SPAN></FONT><SPAN class=yiv639626477jrnl><FONT color=black size=1 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 9pt">Trans R Soc </SPAN></FONT></SPAN><SPAN class=yiv639626477jrnl><FONT color=#1f497d size=1 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: #1f497d; FONT-SIZE: 9pt">Trop Med Hyg</SPAN></FONT></SPAN><FONT color=#1f497d size=1 face=Arial><SPAN style="FONT-FAMILY:
Arial; COLOR: #1f497d; FONT-SIZE: 9pt">. 2008 Jan;102(1):54-7.). We have analyzed several virulence genes comparing the isolates from the diverse clinical forms and they turned to be are highly homologous. Immunologically they induce different immune responses (TH1/TH2). We know the vector of L.donovani VL isolates which is P. orientalis, but we still don’t know the vector of cutaneous L. donovani parasite. We believe the vector plays an important role in the pathogenesis and the outcome of L. donovani infection together with the host immune response and may be some of the parasite virulence factors. We will soon published our data on parasite typing and host immune responses of the diverse clinical forms. </SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=#1f497d size=1 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: #1f497d; FONT-SIZE: 9pt">Best regards</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=#1f497d size=1 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: #1f497d; FONT-SIZE: 9pt">Maowia</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt; FONT-WEIGHT: bold">Professor Maowia M. Mukhtar</SPAN></FONT></B><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt; FONT-WEIGHT: bold">University</SPAN></FONT></B><B><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt; FONT-WEIGHT: bold"> of Khartoum</SPAN></FONT></B><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=#1f497d size=2 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: #1f497d; FONT-SIZE: 11pt; FONT-WEIGHT: bold">P.O. Box 11463, khartoum, Sudan</SPAN></FONT></B><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">From:</SPAN></FONT></B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt"> leish-l-bounces@lineu.icb.usp.br [mailto:leish-l-bounces@lineu.icb.usp.br] <B><SPAN style="FONT-WEIGHT: bold">On Behalf Of </SPAN></B>Sharman<BR><B><SPAN style="FONT-WEIGHT: bold">Sent:</SPAN></B> 31 May 2011 18:13<BR><B><SPAN style="FONT-WEIGHT: bold">To:</SPAN></B> Petr Volf; Chang, Kwang-Poo; Hiro Goto; elfadil abass; vishwamohan_katoch@yahoo.co.in; lalit Kant ICMR<BR><B><SPAN style="FONT-WEIGHT: bold">Cc:</SPAN></B> leish-l@lineu.icb.usp.br<BR><B><SPAN style="FONT-WEIGHT: bold">Subject:</SPAN></B> Re: [Leish-l] inquiry</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=3 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: black; FONT-SIZE: 12pt">Hi Petr,</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=3 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: black; FONT-SIZE: 12pt">Thanks for enclosing the pdf. In fact the strain which we are dealing with is not very difficult to grow but difficult to maintain, It is rK 39 positive and as I mentioned in the conversation below that even the serous exudate from the ulcer is also positive for rK 39 dipstick. In between because of my relocation to a different institute I was little away from this work but I intend to resume this work in couple of months. Although in past we tried to isolate the parasite from sand flies but did not succeed. We will surely attempt again with the techniques described by you and others. Contamination is the problem.</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=3 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: black; FONT-SIZE: 12pt">This particular focus seems to be a complex one.</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=3 face=Calibri><SPAN style="FONT-FAMILY: Calibri; COLOR: black; FONT-SIZE: 12pt">NL Sharma</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"> </SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: whitesmoke" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">From:</SPAN></FONT></B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt"> <A title=volf@cesnet.cz href="mailto:volf@cesnet.cz" rel=nofollow target=_blank ymailto="mailto:volf@cesnet.cz">Petr Volf</A> </SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: whitesmoke" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">Sent:</SPAN></FONT></B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt"> Tuesday, May 31, 2011 12:36 PM</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: whitesmoke" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">To:</SPAN></FONT></B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt"> <A title=KwangPoo.Chang@rosalindfranklin.edu href="mailto:KwangPoo.Chang@rosalindfranklin.edu" rel=nofollow target=_blank ymailto="mailto:KwangPoo.Chang@rosalindfranklin.edu">Chang, Kwang-Poo</A> ; <A title="mailto:nandlals@hotmail.com CTRL + Click to follow link" href="mailto:nandlals@hotmail.com" rel=nofollow target=_blank ymailto="mailto:nandlals@hotmail.com">Sharman</A> ; <A title="mailto:hgoto@usp.br CTRL + Click to follow link" href="mailto:hgoto@usp.br" rel=nofollow target=_blank ymailto="mailto:hgoto@usp.br">Hiro Goto</A> ; <A title="mailto:elfadil_abass@yahoo.com CTRL + Click to follow link"
href="mailto:elfadil_abass@yahoo.com" rel=nofollow target=_blank ymailto="mailto:elfadil_abass@yahoo.com">elfadil abass</A> </SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: whitesmoke" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">Cc:</SPAN></FONT></B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt"> <A title=leish-l@lineu.icb.usp.br href="mailto:leish-l@lineu.icb.usp.br" rel=nofollow target=_blank ymailto="mailto:leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</A> </SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: whitesmoke" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">Subject:</SPAN></FONT></B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt"> Re: [Leish-l] inquiry</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt">Hi K.P.,</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt">in Cukurova region, Turkey , cutaneous L. infantum (now it seems that we are dealing with L.donovani/L. infantum hybrid) grew very poorely if we isolated them from patients. Only 1 of 25 isolations was succesfull. However, the same strain (confirmed by molecular methods) grew repeatedly and very well if we isolated them from sandflies. It might be useful for Dr. Sharma try to get isolates from sand flies. It is very laborious but very useful, in Cukurova we got about dozen of isolates by this method (all identical).</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt">Patients are rK39 negative, see attached paper.</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt">Best wishes</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt">Petr</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt">----- Original Message ----- </SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: #e4e4e4" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">From:</SPAN></FONT></B><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt"> <A title=KwangPoo.Chang@rosalindfranklin.edu href="mailto:KwangPoo.Chang@rosalindfranklin.edu" rel=nofollow target=_blank ymailto="mailto:KwangPoo.Chang@rosalindfranklin.edu">Chang, Kwang-Poo</A> </SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">To:</SPAN></FONT></B><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt"> <A title=nandlals@hotmail.com href="mailto:nandlals@hotmail.com" rel=nofollow target=_blank ymailto="mailto:nandlals@hotmail.com">Sharman</A> ; <A title=hgoto@usp.br href="mailto:hgoto@usp.br" rel=nofollow target=_blank ymailto="mailto:hgoto@usp.br">Hiro Goto</A> ; <A title=elfadil_abass@yahoo.com href="mailto:elfadil_abass@yahoo.com" rel=nofollow target=_blank ymailto="mailto:elfadil_abass@yahoo.com">elfadil abass</A> </SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">Cc:</SPAN></FONT></B><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt"> <A title=leish-l@lineu.icb.usp.br href="mailto:leish-l@lineu.icb.usp.br" rel=nofollow target=_blank ymailto="mailto:leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</A> </SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">Sent:</SPAN></FONT></B><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt"> Sunday, May 22, 2011 1:39 AM</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt">Dr. NL Sharma has been working on an important CL endemic area along the Satluj River valley to the south of Himalaya in India (Please correct me should I be wrong for anything I said here and below). I had the good fortune of visiting the site several years back courtesy of Dr. Sharma. </SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt">I believe Dr. Sharma's finding is important, since the parasites there are different from the familiar Indian L donovani in Bihar . The parasites are refratory to in vitro cultivation. They do differentiate into promastigotes and may grow a little, but can't really be subcultured to establish passageable lines. This is very much reminescent of L infantum in the Mediterranean area. I recall Dr. Sharma has also found rK39+ dogs, if I remember correctly. If so, Satluj river valley is endemic to the infantile CL.</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 10pt">Analyses of several batches of basically clinical CL samples from Dr. Sharma showed evidence of L infantum, but also L tropica as well as a mixture of the two in one sample. This is the same picture we have noted for samples from Hatay , Turkey .</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=3 face=Arial><SPAN style="FONT-FAMILY: Arial; COLOR: black; FONT-SIZE: 12pt">These observations make me wonder a lot about our current knowledge on the clinico-epidemiology based on data collected previously by analyses of cultured promastigotes from one or few 'representative samples'. Nowaday, technology makes it very doable to work with biological samples for Leish DNAs directly from sand flies, patients and reservoir animals.</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt; FONT-WEIGHT: bold">From:</SPAN></FONT></B><FONT color=black size=2 face=Tahoma><SPAN style="FONT-FAMILY: Tahoma; COLOR: black; FONT-SIZE: 10pt"> leish-l-bounces@lineu.icb.usp.br on behalf of Sharman<BR><B><SPAN style="FONT-WEIGHT: bold">Sent:</SPAN></B> Fri 5/20/2011 11:31 PM<BR><B><SPAN style="FONT-WEIGHT: bold">To:</SPAN></B> Hiro Goto; elfadil abass<BR><B><SPAN style="FONT-WEIGHT: bold">Cc:</SPAN></B> leish-l@lineu.icb.usp.br<BR><B><SPAN style="FONT-WEIGHT: bold">Subject:</SPAN></B> Re: [Leish-l] inquiry</SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV></DIV>
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<div style="BACKGROUND: white" class=yiv639626477MsoNormal><FONT color=black size=2 face="Times New Roman"><SPAN style="COLOR: black; FONT-SIZE: 10pt">Dear all<BR>I agree with Hiro gito, We are working on a focus where the CL is<BR>predominantly caused by L. donovani, and the rK 39 STRIPS GIVE POSITIVE<BR>RESULTS WITH SERA as well as serous exudate from the lesion. The results are<BR>dependent upon species.<BR>NL Sharma<BR><BR>--------------------------------------------------<BR>From: "Hiro Goto" <hgoto@usp.br><BR>Sent: Wednesday, May 18, 2011 12:09 AM<BR>To: "elfadil abass" <elfadil_abass@yahoo.com><BR>Cc: <leish-l@lineu.icb.usp.br><BR>Subject: Re: [Leish-l] inquiry<BR><BR>> Dear all,<BR>> In our oppinion, DAT and rK39 for those samples are not indicated since<BR>> these tests are produced for the diagnosis of visceral leishmaniasis.<BR>> In case of tegumentary leishmaniasis, it is very appropriate the<BR>>
observation of J.J. Shaw appointing species specificity of antibody<BR>> response in these cases. Titers of antibodies are in general low in these<BR>> cases therefore depending on the species, it may result negative. We have<BR>> published a review recently in Expert Rev. Anti Infect. Ther. 8(4),<BR>> 419?433 (2010), Current diagnosis and treatment of cutaneous and<BR>> mucocutaneous leishmaniasis, where we raise this point.<BR>> Hiro Goto<BR>><BR>> Citando elfadil abass <elfadil_abass@yahoo.com>:<BR>><BR>>> Dear all I would recommend using DAT and rK39 strip test to measure<BR>>> antibody<BR>>> responses and to evaluate the diagnostic efficiency for both tests in<BR>>> such group<BR>>> of patients.<BR>>><BR>>> Elfadil Abass<BR>>> ________________________________________________________________________________<BR>>> Institute of
Medical Microbiology and Hospital Hygiene<BR>>> Philipps University Marburg<BR>>> BMFZ / Hans-Meerwein Straße 2<BR>>> D-35033 Marburg , Germany<BR>>> ________________________________________________________________________________<BR>>><BR>>><BR>>><BR>>><BR>>> ________________________________<BR>>> From: Nuha Nuwayri-Salti <racha@aub.edu.lb><BR>>> To: saad saad <saad1426@gmail.com>; "leish-l@lineu.icb.usp.br"<BR>>> <leish-l@lineu.icb.usp.br><BR>>> Sent: Fri, May 13, 2011 9:39:33 AM<BR>>> Subject: Re: [Leish-l] inquiry<BR>>><BR>>> Dear Saad first precaution to take is to separate your samples into<BR>>> several<BR>>> portions each (at least 5 each being no more than a few hundred(200-400)<BR>>> microliters). This is a necessary precaution to avoid freezing and<BR>>> thawing<BR>>> several times the same sample
which will be the case should you do<BR>>> different<BR>>> studies at different times which is unavoidable.<BR>>><BR>>> What you can do is correlate the type(ulcerated, abscess, furuncle<BR>>> etc.. ) the<BR>>> locale, the number and age of lesions with the levels of antibody in the<BR>>> sera of<BR>>> these patgients and also monitor cell mediated immunity with leishmanin<BR>>> skin<BR>>> test!<BR>>> I have just published (in print) an article about having circulating<BR>>> parasites<BR>>> in some of these patients with apparently pure cutaneous disease. It is<BR>>> the<BR>>> first paper that revealed this fact. It would be interesting to confirm<BR>>> or<BR>>> de-confirm this fact repeating what we did.<BR>>> Best wishes<BR>>><BR>>> Nuha Nuwayri-Salti MD<BR>>><BR>>><BR>>><BR>>> AOA<BR>>>
Medical Honor Society<BR>>><BR>>><BR>>><BR>>><BR>>> -----Original Message-----<BR>>> From: leish-l-bounces@lineu.icb.usp.br<BR>>> [<A href="mailto:leish-l-bounces@lineu.icb.usp.br" rel=nofollow target=_blank ymailto="mailto:leish-l-bounces@lineu.icb.usp.br">mailto:leish-l-bounces@lineu.icb.usp.br</A>]<BR>>> On Behalf Of saad saad<BR>>> Sent: Friday, April 29, 2011 11:23 PM<BR>>> To: leish-l@lineu.icb.usp.br<BR>>> Subject: [Leish-l] inquiry<BR>>><BR>>> Dear all<BR>>> Hi. i am Saad from Saudi Arabia . i have 60 sera from positive case of<BR>>> CL from south west of the country. I would like to have your<BR>>> recommendations to start a good research line in CL using these sera.<BR>>> Thanks in advance for your help<BR>>> _______________________________________________<BR>>> Leish-l mailing list<BR>>> Leish-l@lineu.icb.usp.br<BR>>>
http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l<BR>>> _______________________________________________<BR>>> Leish-l mailing list<BR>>> Leish-l@lineu.icb.usp.br<BR>>> <A href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l" rel=nofollow target=_blank>http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l</A><BR>>><BR>><BR>><BR>><BR>> Profa. Dra. Hiro Goto<BR>> Laboratório de Soroepidemiologia e Imunobiologia<BR>> Instituto de Medicina Tropical de São Paulo , USP<BR>> Av. Dr. Enéas de Carvalho Aguiar, 470, prédio II, quarto andar<BR>> 05403-000 - São Paulo , SP<BR>> Tel. +55-11-3061 7023, 3061 7056 ou 3061 7027<BR>> Fax. +55-11-3061-8270<BR>><BR>> _______________________________________________<BR>> Leish-l mailing list<BR>> Leish-l@lineu.icb.usp.br<BR>> <A href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l" rel=nofollow
target=_blank>http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l</A><BR>><BR>_______________________________________________<BR>Leish-l mailing list<BR>Leish-l@lineu.icb.usp.br<BR><A href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l" rel=nofollow target=_blank>http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l</A></SPAN></FONT><FONT color=black><SPAN style="COLOR: black"></SPAN></FONT></div></DIV></DIV>
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