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<div class=Section1>
<p class=MsoNormal><font size=2 color=navy face=Arial><span style='font-size:
10.0pt;font-family:Arial;color:navy'>I want to reiterate that L tropica sequence
is also present among the same batch of samples received from Dr. NL Sharma;
and that one sample appears to be a tropica/infantum mixture or hybrid. These
samples are apparently independent of those subsequently sent to you by Dr. Sharma,
since permanent cultures have not been established for investigation by
different groups.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 color=navy face=Arial><span style='font-size:
10.0pt;font-family:Arial;color:navy'><o:p> </o:p></span></font></p>
<p class=MsoNormal><font size=2 color=navy face=Arial><span style='font-size:
10.0pt;font-family:Arial;color:navy'>It is good to know that you have secured
funding to continue your collaborative research of importance with Dr. Sharma. <o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 color=navy face=Arial><span style='font-size:
10.0pt;font-family:Arial;color:navy'><o:p> </o:p></span></font></p>
<p class=MsoNormal><font size=2 color=navy face=Arial><span style='font-size:
10.0pt;font-family:Arial;color:navy'>KP<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 color=navy face=Arial><span style='font-size:
10.0pt;font-family:Arial;color:navy'><o:p> </o:p></span></font></p>
<div>
<div class=MsoNormal align=center style='text-align:center'><font size=3
face="Times New Roman"><span style='font-size:12.0pt'>
<hr size=2 width="100%" align=center tabindex=-1>
</span></font></div>
<p class=MsoNormal><b><font size=2 face=Tahoma><span style='font-size:10.0pt;
font-family:Tahoma;font-weight:bold'>From:</span></font></b><font size=2
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma'> <st1:PersonName
w:st="on">Dr.Sunil Arora</st1:PersonName> [mailto:skarora_in@yahoo.com] <br>
<b><span style='font-weight:bold'>Sent:</span></b> Friday, June 03, 2011 11:36
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><o:p></o:p></p>
</div>
<p class=MsoNormal><font size=3 face="Times New Roman"><span style='font-size:
12.0pt'><o:p> </o:p></span></font></p>
<div>
<div style='right:auto'>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'>Dear
KP</span><o:p></o:p></span></font></p>
</div>
<span style='right:auto'>
<div style='right:auto'>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'>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><o:p></o:p></span></font></p>
</div>
<div style='right:auto'>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'> <o:p></o:p></span></font></p>
</div>
<span style='right:auto'>
<div style='right:auto'>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'>anyways...
very nice to have your exclusive comments and suggestions</span><o:p></o:p></span></font></p>
</div>
<span style='right:auto'>
<div style='right:auto'>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'>regards</span><o:p></o:p></span></font></p>
</div>
<span style='right:auto'>
<div style='right:auto'>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'>sunil</span><o:p></o:p></span></font></p>
</div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'> <o:p></o:p></span></font></p>
</div>
<div style='right:auto'>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'> <o:p></o:p></span></font></p>
</div>
<div>
<div>
<p class=MsoNormal style='margin-bottom:12.0pt;background:white'><b><font
size=2 color=black face=Arial><span style='font-size:10.0pt;font-family:Arial;
color:black;font-weight:bold'>From:</span></font></b><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black'>
"Chang, Kwang-Poo" <KwangPoo.Chang@rosalindfranklin.edu><br>
<b><span style='font-weight:bold'>To:</span></b> <st1:PersonName w:st="on">Dr.Sunil
Arora</st1:PersonName> <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</span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
<div id=yiv639626477>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'>Dear Sunil,</span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'>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><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
<ol start=1 type=1>
<li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:
auto;mso-list:l7 level1 lfo1;background:white'><font size=2 color=navy
face=Arial><span style='font-size:10.0pt;font-family:Arial'>It is
important for you and Dr. Sharma to work together, which is very much in
line with the spirit of WorldLeish.</span></font><o:p></o:p></li>
</ol>
<div style='margin-left:.25in'>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
<ol start=2 type=1>
<li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:
auto;mso-list:l4 level1 lfo2;background:white'><font size=2 color=navy
face=Arial><span style='font-size:10.0pt;font-family:Arial'>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 <st1:place w:st="on">Bihar</st1:place>
. Splenic aspirates from <st1:PersonName w:st="on">Shyam Sundar</st1:PersonName>’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 <st1:country-region w:st="on">China</st1:country-region>
(Xinjiang, <st1:City w:st="on">Beijing</st1:City> , <st1:State w:st="on">Shandong</st1:State>
, <st1:State w:st="on">Sichuan</st1:State> , <st1:State w:st="on">Gansu</st1:State>
), <st1:country-region w:st="on">Pakistan</st1:country-region> , <st1:country-region
w:st="on">Iran</st1:country-region> , <st1:country-region w:st="on"><st1:place
w:st="on">Turkey</st1:place></st1:country-region> 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, <st1:PersonName w:st="on">Shyam Sundar</st1:PersonName>, JC
Dujardin and their colleagues recently reported the isolation of
Leptomonas from ~7% (according to Shyam) of his <st1:place w:st="on">Bihar</st1:place>
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><o:p></o:p></li>
</ol>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
<ol start=3 type=1>
<li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:
auto;mso-list:l6 level1 lfo3;background:white'><font size=2 color=navy
face=Arial><span style='font-size:10.0pt;font-family:Arial'>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><o:p></o:p></li>
</ol>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
<ol start=4 type=1>
<li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:
auto;mso-list:l5 level1 lfo4;background:white'><font size=2 color=navy
face=Arial><span style='font-size:10.0pt;font-family:Arial'>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><o:p></o:p></li>
</ol>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
<ol start=5 type=1>
<li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:
auto;mso-list:l2 level1 lfo5;background:white'><font size=2 color=navy
face=Arial><span style='font-size:10.0pt;font-family:Arial'>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. <st1:PersonName w:st="on">Carlos
Costa</st1:PersonName> also told me that he has no difficulty of growing
L chagasi from his patients in <st1:City w:st="on"><st1:place w:st="on">Teresina</st1:place></st1:City>
. 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><o:p></o:p></li>
</ol>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'>KP</span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><b><font size=2 color=black
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma;color:black;
font-weight:bold'>From:</span></font></b><font size=2 color=black face=Tahoma><span
style='font-size:10.0pt;font-family:Tahoma;color:black'> <st1:PersonName w:st="on">Dr.Sunil
Arora</st1:PersonName> [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><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<p class=MsoNormal style='background:white'><font size=3 color=black
face="Times New Roman"><span style='font-size:12.0pt;color:black'> <o:p></o:p></span></font></p>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'>Dear
KP and all</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'>I
have been reading the exchange of emails regarding characterisation of
leishmania isolates from Himachal in <st1:country-region w:st="on"><st1:place
w:st="on">India</st1:place></st1:country-region> 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 <st1:country-region w:st="on"><st1:place w:st="on">India</st1:place></st1:country-region>
. 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></font><font color=black><span
style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'>regards</span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'>sunil</span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Verdana><span style='font-size:10.0pt;font-family:Verdana;color:black'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><span class=msid8447><i><font
size=2 color="#00007f" face=Verdana><span style='font-size:10.0pt;font-family:
Verdana;color:#00007F;font-style:italic'>Dr Sunil K.Arora</span></font></i></span><i><font
size=2 color="#00007f" face=Verdana><span style='font-size:10.0pt;font-family:
Verdana;color:#00007F;font-style:italic'><br>
<span class=msid8451>Professor </span><br>
<span class=msid8453>In Charge HIV diagnostic and Disease Monitoring Laboratory</span><br>
<span class=msid8455>Department of Immunopathology</span><br>
<span class=msid8457>PGIMER, Chandigarh-160 012</span><br>
<span class=msid8459>Ph.: 0091-172-2755192(Off)</span><br>
<span class=msid8461>FAX: 0091-172-2744401, 2745078</span><br>
<span class=msid8463>Ph:+91-172-2732863 (Res); 9872866087 (cell)</span><br>
<span class=msid8465>email: skarora_in@yahoo.com; skarorain@gmail.com</span></span></font></i><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<div style='margin-bottom:12.0pt'>
<p class=MsoNormal style='background:white'><b><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black;
font-weight:bold'>From:</span></font></b><font size=2 color=black face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:black'> "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'><o:p></o:p></span></font></p>
</div>
<div id=yiv639626477>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'>Dear Maowia,</span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'>Thanks again for the
info.</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'>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'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'>KP</span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><b><font size=2 color=black
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma;color:black;
font-weight:bold'>From:</span></font></b><font size=2 color=black face=Tahoma><span
style='font-size:10.0pt;font-family:Tahoma;color:black'> 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'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=3 color=black
face="Times New Roman"><span style='font-size:12.0pt;color:black'> <o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'>Dear
KP,</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'>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 <st1:country-region
w:st="on"><st1:place w:st="on">Sudan</st1:place></st1:country-region> 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'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'>Best
regards</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'>Maowia</span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><b><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D;
font-weight:bold'>Professor Maowia M. Mukhtar</span></font></b><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><st1:place w:st="on"><st1:PlaceType
w:st="on"><b><font size=2 color="#1f497d" face=Calibri><span style='font-size:
11.0pt;font-family:Calibri;color:#1F497D;font-weight:bold'>Institute</span></font></b></st1:PlaceType><b><font
size=2 color="#1f497d" face=Calibri><span style='font-size:11.0pt;font-family:
Calibri;color:#1F497D;font-weight:bold'> of <st1:PlaceName w:st="on">Endemic</st1:PlaceName></span></font></b></st1:place><b><font
size=2 color="#1f497d" face=Calibri><span style='font-size:11.0pt;font-family:
Calibri;color:#1F497D;font-weight:bold'> Diseases</span></font></b><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><st1:place w:st="on"><st1:PlaceType
w:st="on"><b><font size=2 color="#1f497d" face=Calibri><span style='font-size:
11.0pt;font-family:Calibri;color:#1F497D;font-weight:bold'>University</span></font></b></st1:PlaceType><b><font
size=2 color="#1f497d" face=Calibri><span style='font-size:11.0pt;font-family:
Calibri;color:#1F497D;font-weight:bold'> of <st1:PlaceName w:st="on">Khartoum</st1:PlaceName></span></font></b></st1:place><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><b><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D;
font-weight:bold'>P.O. Box 11463, khartoum, Sudan</span></font></b><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><st1:City w:st="on"><st1:place
w:st="on"><b><font size=2 color="#1f497d" face=Calibri><span style='font-size:
11.0pt;font-family:Calibri;color:#1F497D;font-weight:bold'>Mobile</span></font></b></st1:place></st1:City><b><font
size=2 color="#1f497d" face=Calibri><span style='font-size:11.0pt;font-family:
Calibri;color:#1F497D;font-weight:bold'>; +249912234268</span></font></b><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><b><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D;
font-weight:bold'>Fax: +249183779712</span></font></b><font color=black><span
style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div style='border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0in 0in 0in'>
<div>
<div>
<p class=MsoNormal style='background:white'><b><font size=2 color=black
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma;color:black;
font-weight:bold'>From:</span></font></b><font size=2 color=black face=Tahoma><span
style='font-size:10.0pt;font-family:Tahoma;color:black'> 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'><o:p></o:p></span></font></p>
</div>
</div>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=3 color=black
face="Times New Roman"><span style='font-size:12.0pt;color:black'> <o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'>Dear Maowia,</span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'>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'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<ol style='margin-top:0in' start=1 type=1>
<li class=MsoNormal style='color:navy;mso-list:l8 level1 lfo6;background:white'><font
size=2 color=navy face=Arial><span style='font-size:10.0pt;font-family:
Arial'>“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><o:p></o:p></li>
</ol>
<div style='margin-left:.25in'>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<ol style='margin-top:0in' start=2 type=1>
<li class=MsoNormal style='color:navy;mso-list:l3 level1 lfo7;background:white'><font
size=2 color=navy face=Arial><span style='font-size:10.0pt;font-family:
Arial'>“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><o:p></o:p></li>
</ol>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<ol style='margin-top:0in' start=3 type=1>
<li class=MsoNormal style='color:navy;mso-list:l0 level1 lfo8;background:white'><font
size=2 color=navy face=Arial><span style='font-size:10.0pt;font-family:
Arial'>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><o:p></o:p></li>
</ol>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<ol style='margin-top:0in' start=4 type=1>
<li class=MsoNormal style='color:navy;mso-list:l1 level1 lfo9;background:white'><font
size=2 color=navy face=Arial><span style='font-size:10.0pt;font-family:
Arial'>The Sudanese strain used by many in the <st1:country-region w:st="on"><st1:place
w:st="on">USA</st1:place></st1:country-region> 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 <st1:place w:st="on">Bihar</st1:place> 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><o:p></o:p></li>
</ol>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'>KP</span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=navy face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:navy'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><b><font size=2 color=black
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma;color:black;
font-weight:bold'>From:</span></font></b><font size=2 color=black face=Tahoma><span
style='font-size:10.0pt;font-family:Tahoma;color:black'> 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'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=3 color=black
face="Times New Roman"><span style='font-size:12.0pt;color:black'> <o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'>Dear
Colleagues,</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'>We
have very interesting experience with Sudanese L.donovani. This group of
parasite although are genetically homogenous they still cause diverse clinical
forms in <st1:country-region w:st="on"><st1:place w:st="on">Sudan</st1:place></st1:country-region>
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 size=1 color=black face=Arial><span
style='font-size:9.0pt;font-family:Arial;color:black'>Trans R Soc </span></font></span><span
class=yiv639626477jrnl><font size=1 color="#1f497d" face=Arial><span
style='font-size:9.0pt;font-family:Arial;color:#1F497D'>Trop Med Hyg</span></font></span><font
size=1 color="#1f497d" face=Arial><span style='font-size:9.0pt;font-family:
Arial;color:#1F497D'>. 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'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=1 color="#1f497d"
face=Arial><span style='font-size:9.0pt;font-family:Arial;color:#1F497D'>Best
regards</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=1 color="#1f497d"
face=Arial><span style='font-size:9.0pt;font-family:Arial;color:#1F497D'>Maowia</span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><b><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D;
font-weight:bold'>Professor Maowia M. Mukhtar</span></font></b><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><st1:place w:st="on"><st1:PlaceType
w:st="on"><b><font size=2 color="#1f497d" face=Calibri><span style='font-size:
11.0pt;font-family:Calibri;color:#1F497D;font-weight:bold'>Institute</span></font></b></st1:PlaceType><b><font
size=2 color="#1f497d" face=Calibri><span style='font-size:11.0pt;font-family:
Calibri;color:#1F497D;font-weight:bold'> of <st1:PlaceName w:st="on">Endemic</st1:PlaceName></span></font></b></st1:place><b><font
size=2 color="#1f497d" face=Calibri><span style='font-size:11.0pt;font-family:
Calibri;color:#1F497D;font-weight:bold'> Diseases</span></font></b><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><st1:place w:st="on"><st1:PlaceType
w:st="on"><b><font size=2 color="#1f497d" face=Calibri><span style='font-size:
11.0pt;font-family:Calibri;color:#1F497D;font-weight:bold'>University</span></font></b></st1:PlaceType><b><font
size=2 color="#1f497d" face=Calibri><span style='font-size:11.0pt;font-family:
Calibri;color:#1F497D;font-weight:bold'> of <st1:PlaceName w:st="on">Khartoum</st1:PlaceName></span></font></b></st1:place><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><b><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D;
font-weight:bold'>P.O. Box 11463, khartoum, Sudan</span></font></b><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><st1:City w:st="on"><st1:place
w:st="on"><b><font size=2 color="#1f497d" face=Calibri><span style='font-size:
11.0pt;font-family:Calibri;color:#1F497D;font-weight:bold'>Mobile</span></font></b></st1:place></st1:City><b><font
size=2 color="#1f497d" face=Calibri><span style='font-size:11.0pt;font-family:
Calibri;color:#1F497D;font-weight:bold'>; +249912234268</span></font></b><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><b><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D;
font-weight:bold'>Fax: +249183779712</span></font></b><font color=black><span
style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color="#1f497d"
face=Calibri><span style='font-size:11.0pt;font-family:Calibri;color:#1F497D'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
<div>
<div style='border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0in 0in 0in'>
<div>
<div>
<p class=MsoNormal style='background:white'><b><font size=2 color=black
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma;color:black;
font-weight:bold'>From:</span></font></b><font size=2 color=black face=Tahoma><span
style='font-size:10.0pt;font-family:Tahoma;color:black'> 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'><o:p></o:p></span></font></p>
</div>
</div>
</div>
</div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=3 color=black
face="Times New Roman"><span style='font-size:12.0pt;color:black'> <o:p></o:p></span></font></p>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=3 color=black
face=Calibri><span style='font-size:12.0pt;font-family:Calibri;color:black'>Hi
Petr,</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=3 color=black
face=Calibri><span style='font-size:12.0pt;font-family:Calibri;color:black'>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'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=3 color=black
face=Calibri><span style='font-size:12.0pt;font-family:Calibri;color:black'>This
particular focus seems to be a complex one.</span></font><font color=black><span
style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=3 color=black
face=Calibri><span style='font-size:12.0pt;font-family:Calibri;color:black'>NL
Sharma</span></font><font color=black><span style='color:black'> <o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma;color:black'> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<div>
<p class=MsoNormal style='background:whitesmoke'><b><font size=2 color=black
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma;color:black;
font-weight:bold'>From:</span></font></b><font size=2 color=black face=Tahoma><span
style='font-size:10.0pt;font-family:Tahoma;color:black'> <a
href="mailto:volf@cesnet.cz" target="_blank" title="volf@cesnet.cz"
ymailto="mailto:volf@cesnet.cz">Petr Volf</a> </span></font><font color=black><span
style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:whitesmoke'><b><font size=2 color=black
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma;color:black;
font-weight:bold'>Sent:</span></font></b><font size=2 color=black face=Tahoma><span
style='font-size:10.0pt;font-family:Tahoma;color:black'> Tuesday, May 31, 2011
12:36 PM</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:whitesmoke'><b><font size=2 color=black
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma;color:black;
font-weight:bold'>To:</span></font></b><font size=2 color=black face=Tahoma><span
style='font-size:10.0pt;font-family:Tahoma;color:black'> <a
href="mailto:KwangPoo.Chang@rosalindfranklin.edu" target="_blank"
title="KwangPoo.Chang@rosalindfranklin.edu"
ymailto="mailto:KwangPoo.Chang@rosalindfranklin.edu">Chang, Kwang-Poo</a> ; <a
href="mailto:nandlals@hotmail.com" target="_blank"
title="mailto:nandlals@hotmail.com CTRL + Click to follow link"
ymailto="mailto:nandlals@hotmail.com">Sharman</a> ; <a
href="mailto:hgoto@usp.br" target="_blank"
title="mailto:hgoto@usp.br CTRL + Click to follow link"
ymailto="mailto:hgoto@usp.br">Hiro Goto</a> ; <a
href="mailto:elfadil_abass@yahoo.com" target="_blank"
title="mailto:elfadil_abass@yahoo.com CTRL + Click to follow link"
ymailto="mailto:elfadil_abass@yahoo.com">elfadil abass</a> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:whitesmoke'><b><font size=2 color=black
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma;color:black;
font-weight:bold'>Cc:</span></font></b><font size=2 color=black face=Tahoma><span
style='font-size:10.0pt;font-family:Tahoma;color:black'> <a
href="mailto:leish-l@lineu.icb.usp.br" target="_blank"
title="leish-l@lineu.icb.usp.br" ymailto="mailto:leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a>
</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:whitesmoke'><b><font size=2 color=black
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma;color:black;
font-weight:bold'>Subject:</span></font></b><font size=2 color=black
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma;color:black'> Re:
[Leish-l] inquiry</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=3 color=black
face="Times New Roman"><span style='font-size:12.0pt;color:black'> <o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black'>Hi
K.P.,</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black'>in
Cukurova region, <st1:country-region w:st="on"><st1:place w:st="on">Turkey</st1:place></st1:country-region>
, 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'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black'>Patients
are rK39 negative, see attached paper.</span></font><font color=black><span
style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black'>Best
wishes</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black'>Petr</span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<blockquote style='border:none;border-left:solid black 1.5pt;padding:0in 0in 0in 4.0pt;
margin-left:3.75pt;margin-top:5.0pt;margin-right:0in;margin-bottom:5.0pt'>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black'>-----
Original Message ----- </span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:#E4E4E4'><b><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black;
font-weight:bold'>From:</span></font></b><font size=2 color=black face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:black'> <a
href="mailto:KwangPoo.Chang@rosalindfranklin.edu" target="_blank"
title="KwangPoo.Chang@rosalindfranklin.edu"
ymailto="mailto:KwangPoo.Chang@rosalindfranklin.edu">Chang, Kwang-Poo</a> </span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
</div>
</div>
</div>
<div>
<div>
<div>
<p class=MsoNormal style='background:white'><b><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black;
font-weight:bold'>To:</span></font></b><font size=2 color=black face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:black'> <a
href="mailto:nandlals@hotmail.com" target="_blank" title="nandlals@hotmail.com"
ymailto="mailto:nandlals@hotmail.com">Sharman</a> ; <a
href="mailto:hgoto@usp.br" target="_blank" title="hgoto@usp.br"
ymailto="mailto:hgoto@usp.br">Hiro Goto</a> ; <a
href="mailto:elfadil_abass@yahoo.com" target="_blank"
title="elfadil_abass@yahoo.com" ymailto="mailto:elfadil_abass@yahoo.com">elfadil
abass</a> </span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
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<p class=MsoNormal style='background:white'><b><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black;
font-weight:bold'>Cc:</span></font></b><font size=2 color=black face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:black'> <a
href="mailto:leish-l@lineu.icb.usp.br" target="_blank"
title="leish-l@lineu.icb.usp.br" ymailto="mailto:leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a>
</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
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<p class=MsoNormal style='background:white'><b><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black;
font-weight:bold'>Sent:</span></font></b><font size=2 color=black face=Arial><span
style='font-size:10.0pt;font-family:Arial;color:black'> Sunday, May 22, 2011
1:39 AM</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
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<p class=MsoNormal style='background:white'><b><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black;
font-weight:bold'>Subject:</span></font></b><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black'> Re:
[Leish-l] inquiry</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
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<p class=MsoNormal style='background:white'><font size=3 color=black
face="Times New Roman"><span style='font-size:12.0pt;color:black'> <o:p></o:p></span></font></p>
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<div id=yiv639626477idOWAReplyText41530>
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<p class=MsoNormal style='background:white'><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black'>Dr. NL
Sharma has been working on an important CL endemic area along the <st1:PlaceName
w:st="on">Satluj</st1:PlaceName> <st1:PlaceType w:st="on">River</st1:PlaceType>
valley to the south of Himalaya in <st1:country-region w:st="on"><st1:place
w:st="on">India</st1:place></st1:country-region> (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'><o:p></o:p></span></font></p>
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<p class=MsoNormal style='background:white'><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black'>I
believe Dr. Sharma's finding is important, since the parasites there are
different from the familiar Indian L donovani in <st1:place w:st="on">Bihar</st1:place>
. 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'><o:p></o:p></span></font></p>
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face="Times New Roman"><span style='font-size:12.0pt;color:black'> <o:p></o:p></span></font></p>
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<p class=MsoNormal style='background:white'><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black'>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 <st1:place
w:st="on"><st1:City w:st="on">Hatay</st1:City> , <st1:country-region w:st="on">Turkey</st1:country-region></st1:place>
.</span></font><font color=black><span style='color:black'><o:p></o:p></span></font></p>
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<p class=MsoNormal style='background:white'><font size=3 color=black
face=Arial><span style='font-size:12.0pt;font-family:Arial;color:black'>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'><o:p></o:p></span></font></p>
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<p class=MsoNormal style='background:white'><font size=2 color=black
face=Arial><span style='font-size:10.0pt;font-family:Arial;color:black'>KP</span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
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<p class=MsoNormal style='background:white'><b><font size=2 color=black
face=Tahoma><span style='font-size:10.0pt;font-family:Tahoma;color:black;
font-weight:bold'>From:</span></font></b><font size=2 color=black face=Tahoma><span
style='font-size:10.0pt;font-family:Tahoma;color:black'>
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'><o:p></o:p></span></font></p>
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<p class=MsoNormal style='background:white'><font size=2 color=black
face="Times New Roman"><span style='font-size:10.0pt;color:black'>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>
>> <st1:place w:st="on"><st1:PlaceType w:st="on">Institute</st1:PlaceType>
of <st1:PlaceName w:st="on">Medical</st1:PlaceName></st1:place> Microbiology
and Hospital Hygiene<br>
>> <st1:PlaceName w:st="on">Philipps</st1:PlaceName> <st1:PlaceName
w:st="on">University</st1:PlaceName> <st1:City w:st="on"><st1:place w:st="on">Marburg</st1:place></st1:City><br>
>> BMFZ / Hans-Meerwein Straße 2<br>
>> D-35033 <st1:place w:st="on"><st1:City w:st="on">Marburg</st1:City> , <st1:country-region
w:st="on">Germany</st1:country-region></st1:place><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" 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 <st1:country-region w:st="on"><st1:place w:st="on">Saudi
Arabia</st1:place></st1:country-region> . 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"
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 <st1:City w:st="on"><st1:place w:st="on">São
Paulo</st1:place></st1:City> , USP<br>
> Av. Dr. Enéas de Carvalho Aguiar, 470, prédio II, quarto andar<br>
> 05403-000 - <st1:City w:st="on"><st1:place w:st="on">São Paulo</st1:place></st1:City>
, 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"
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"
target="_blank">http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l</a></span></font><font
color=black><span style='color:black'><o:p></o:p></span></font></p>
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<p class=MsoNormal style='background:white'><font size=3 color=black
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_______________________________________________<br>
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<a href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l"
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