<html><head></head><body style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; ">Dear All,<div>1- Try taking a few Balb/c mice to the field and inject your specimens, depending on the suspected organism, to different sites i.e. ear, i.v., id or sc. Harvest a week or several weeks later from the site of injection, draining lymph node or spleen. Usually, contaminant will not survive.</div><div>2- Patrick Bastien mentioned an important point and said he does not want to start a discussion - what a pity! The point is that neither isoenzyme nor the genetic markers that have been used for identification of <i>Leishmania</i> are biological markers. They have been very useful tools to study evolution and inter-relations of different species, but not defining necessarily the disease they produce (their biology). All the variants of an isoenzyme have the same enzymatic activity only somewhere else in the molecule a change occurred that makes it migrate differently, but does not grossly change its enzymatic activity. Molecular markers used so far are not biological markers either. Of course if parasites are closely related they would tend to share many biological markers as well. This fact has caused a confusion and many take non-biological markers to define the biology of a parasite: <i>L. donovani</i> that causes CL, or <i>L. tropic</i> that grows and produces a lesion in mice! The biological markers to differentiate between species are few and ancient and no one is too keen to use them, rightly because they are somewhat inaccurate and cumbersome. They were based on epidemiology (anthroponotic vs zoonotic), the general form of disease they produced, predominant vector and reservoir, etc. <i>L. tropic</i> is defined by being anthoponotic, transmitted primarily by <i>P.</i> <i>sergenti</i>, would not produce CL in mice, and the CL it produces in humans tend to be the dry type (not in all cases), and takes a longer time to self cure (over a year in general). As opposed to <i>L. major</i>, that would produce CL (and or VL) in mice i.e. Balb/c, is zoonotic, transmitted generally by <i>P. papatasi</i> or a few others but not <i>sergenti</i> lesions are generally the wet type (not all), and tend to heal in a few months. </div><div>I hope we would have some efforts towards using genomics and other modern tools for identifying biological markers.</div><div>Thanks,</div><div>Farrokh Modabber </div><div><br></div><div><br></div><div> <br><div><div>On Jun 2, 2011, at 8:33 PM, Chang, Kwang-Poo wrote:</div><br class="Apple-interchange-newline"><blockquote type="cite"><o:smarttagtype namespaceuri="urn:schemas-microsoft-com:office:smarttags" name="City">
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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">Dear Maowia,<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">Thanks again for the info.<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">Are there any record available for the
number of successful isolations versus the total number attempted for a given
endemic site ?<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"> Maowia Mukhtar
[mailto:<st1:personname w:st="on"><a href="mailto:mmukhtar@tropmedicine.org">mmukhtar@tropmedicine.org</a></st1:personname>] <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'; <a href="mailto:vishwamohan_katoch@yahoo.co.in">vishwamohan_katoch@yahoo.co.in</a>;
'lalit Kant ICMR'<br>
<b><span style="font-weight:bold">Cc:</span></b> <a href="mailto:leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a><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><p class="MsoNormal"><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D">Dear KP,<o:p></o:p></span></font></p><p class="MsoNormal"><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. <o:p></o:p></span></font></p><p class="MsoNormal"><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D">Best regards<o:p></o:p></span></font></p><p class="MsoNormal"><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D">Maowia<o:p></o:p></span></font></p><p class="MsoNormal"><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D"><o:p> </o:p></span></font></p>
<div><p class="MsoNormal"><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D"><o:p> </o:p></span></font></p><p class="MsoNormal"><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D"><o:p> </o:p></span></font></p><p class="MsoNormal"><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<o:p></o:p></span></font></b></p><p class="MsoNormal"><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<o:p></o:p></span></font></b></p><p class="MsoNormal"><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><b><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:
Calibri;color:#1F497D;font-weight:bold"><o:p></o:p></span></font></b></p><p class="MsoNormal"><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<o:p></o:p></span></font></b></p><p class="MsoNormal"><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<o:p></o:p></span></font></b></p><p class="MsoNormal"><b><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D;font-weight:bold">Fax:
+249183779712<o:p></o:p></span></font></b></p>
</div><p class="MsoNormal"><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D"><o:p> </o:p></span></font></p>
<div>
<div style="border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0in 0in 0in"><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"> 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; <a href="mailto:vishwamohan_katoch@yahoo.co.in">vishwamohan_katoch@yahoo.co.in</a>; lalit Kant ICMR<br>
<b><span style="font-weight:bold">Cc:</span></b> <a href="mailto:leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a>;
Chang, Kwang-Poo<br>
<b><span style="font-weight:bold">Subject:</span></b> RE: [Leish-l] inquiry<o:p></o:p></span></font></p>
</div>
</div><p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:
12.0pt"><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">Dear Maowia,<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">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.<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>
<ol style="margin-top:0in" start="1" type="1">
<li class="MsoNormal" style="color:navy;mso-list:l0 level1 lfo3"><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 ?<o:p></o:p></span></font></li>
</ol><p class="MsoNormal" style="margin-left:.25in"><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>
<ol style="margin-top:0in" start="2" type="1">
<li class="MsoNormal" style="color:navy;mso-list:l0 level1 lfo3"><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 ?<o:p></o:p></span></font></li>
</ol><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>
<ol style="margin-top:0in" start="3" type="1">
<li class="MsoNormal" style="color:navy;mso-list:l0 level1 lfo3"><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 ? <o:p></o:p></span></font></li>
</ol><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>
<ol style="margin-top:0in" start="4" type="1">
<li class="MsoNormal" style="color:navy;mso-list:l0 level1 lfo3"><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. <o:p></o:p></span></font></li>
</ol><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><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">
</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"> Maowia Mukhtar
[mailto:<st1:personname w:st="on"><a href="mailto:mmukhtar@tropmedicine.org">mmukhtar@tropmedicine.org</a></st1:personname>] <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'; <a href="mailto:vishwamohan_katoch@yahoo.co.in">vishwamohan_katoch@yahoo.co.in</a>; 'lalit
Kant ICMR'<br>
<b><span style="font-weight:bold">Cc:</span></b> <a href="mailto:leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a><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><p class="MsoNormal"><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D">Dear Colleagues,<o:p></o:p></span></font></p><p class="MsoNormal"><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="jrnl"><font size="1" face="Arial"><span style="font-size:9.0pt;font-family:
Arial">Trans R Soc <font color="#1f497d"><span style="color:#1F497D">Trop Med
Hyg</span></font></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. <o:p></o:p></span></font></p><p class="MsoNormal"><font size="1" color="#1f497d" face="Arial"><span style="font-size:9.0pt;font-family:Arial;color:#1F497D">Best regards<o:p></o:p></span></font></p><p class="MsoNormal"><font size="1" color="#1f497d" face="Arial"><span style="font-size:9.0pt;font-family:Arial;color:#1F497D">Maowia</span></font><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:
Calibri;color:#1F497D"><o:p></o:p></span></font></p><p class="MsoNormal"><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D"><o:p> </o:p></span></font></p>
<div><p class="MsoNormal"><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D"><o:p> </o:p></span></font></p><p class="MsoNormal"><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D"><o:p> </o:p></span></font></p><p class="MsoNormal"><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<o:p></o:p></span></font></b></p><p class="MsoNormal"><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<o:p></o:p></span></font></b></p><p class="MsoNormal"><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><b><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:
Calibri;color:#1F497D;font-weight:bold"><o:p></o:p></span></font></b></p><p class="MsoNormal"><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<o:p></o:p></span></font></b></p><p class="MsoNormal"><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<o:p></o:p></span></font></b></p><p class="MsoNormal"><b><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D;font-weight:bold">Fax:
+249183779712<o:p></o:p></span></font></b></p>
</div><p class="MsoNormal"><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D"><o:p> </o:p></span></font></p>
<div>
<div style="border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0in 0in 0in"><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">
<a href="mailto:leish-l-bounces@lineu.icb.usp.br">leish-l-bounces@lineu.icb.usp.br</a> [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; <a href="mailto:vishwamohan_katoch@yahoo.co.in">vishwamohan_katoch@yahoo.co.in</a>; lalit Kant ICMR<br>
<b><span style="font-weight:bold">Cc:</span></b> <a href="mailto:leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a><br>
<b><span style="font-weight:bold">Subject:</span></b> Re: [Leish-l] inquiry<o:p></o:p></span></font></p>
</div>
</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><p class="MsoNormal"><font size="3" face="Calibri"><span style="font-size:12.0pt;
font-family:Calibri">Hi Petr,</span></font><o:p></o:p></p>
</div>
<div><p class="MsoNormal"><font size="3" face="Calibri"><span style="font-size:12.0pt;
font-family:Calibri">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><o:p></o:p></p>
</div>
<div><p class="MsoNormal"><font size="3" face="Calibri"><span style="font-size:12.0pt;
font-family:Calibri">This particular focus seems to be a complex one.</span></font><o:p></o:p></p>
</div>
<div><p class="MsoNormal"><font size="3" face="Calibri"><span style="font-size:12.0pt;
font-family:Calibri">NL Sharma</span></font> <o:p></o:p></p>
</div>
<div>
<div><p class="MsoNormal"><font size="2" face="Tahoma"><span style="font-size:10.0pt;
font-family:Tahoma"><o:p> </o:p></span></font></p>
</div>
<div>
<div><p class="MsoNormal" style="background:whitesmoke"><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"> <a href="mailto:volf@cesnet.cz" title="volf@cesnet.cz">Petr Volf</a> <o:p></o:p></span></font></p>
</div>
<div><p class="MsoNormal" style="background:whitesmoke"><b><font size="2" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma;font-weight:bold">Sent:</span></font></b><font size="2" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma"> Tuesday,
May 31, 2011 12:36 PM<o:p></o:p></span></font></p>
</div>
<div><p class="MsoNormal" style="background:whitesmoke"><b><font size="2" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma;font-weight:bold">To:</span></font></b><font size="2" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma"> <a href="mailto:KwangPoo.Chang@rosalindfranklin.edu" title="KwangPoo.Chang@rosalindfranklin.edu">Chang, Kwang-Poo</a> ; <a href="mailto:nandlals@hotmail.com" title="mailto:nandlals@hotmail.com
CTRL + Click to follow link">Sharman</a>
; <a href="mailto:hgoto@usp.br" title="mailto:hgoto@usp.br
CTRL + Click to follow link">Hiro Goto</a> ; <a href="mailto:elfadil_abass@yahoo.com" title="mailto:elfadil_abass@yahoo.com
CTRL + Click to follow link">elfadil
abass</a> <o:p></o:p></span></font></p>
</div>
<div><p class="MsoNormal" style="background:whitesmoke"><b><font size="2" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma;font-weight:bold">Cc:</span></font></b><font size="2" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma"> <a href="mailto:leish-l@lineu.icb.usp.br" title="leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a>
<o:p></o:p></span></font></p>
</div>
<div><p class="MsoNormal" style="background:whitesmoke"><b><font size="2" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma;font-weight:bold">Subject:</span></font></b><font size="2" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma"> Re:
[Leish-l] inquiry<o:p></o:p></span></font></p>
</div>
</div>
</div>
<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><p class="MsoNormal"><font size="2" face="Arial"><span style="font-size:10.0pt;
font-family:Arial">Hi K.P.,</span></font><o:p></o:p></p>
</div>
<div><p class="MsoNormal"><font size="2" face="Arial"><span style="font-size:10.0pt;
font-family:Arial">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><o:p></o:p></p>
</div>
<div><p class="MsoNormal"><font size="2" face="Arial"><span style="font-size:10.0pt;
font-family:Arial">Patients are rK39 negative, see attached paper.</span></font><o:p></o:p></p>
</div>
<div><p class="MsoNormal"><font size="2" face="Arial"><span style="font-size:10.0pt;
font-family:Arial">Best wishes</span></font><o:p></o:p></p>
</div>
<div><p class="MsoNormal"><font size="2" face="Arial"><span style="font-size:10.0pt;
font-family:Arial">Petr</span></font><o:p></o:p></p>
</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><p class="MsoNormal"><font size="2" face="Arial"><span style="font-size:10.0pt;
font-family:Arial">----- Original Message ----- <o:p></o:p></span></font></p>
</div>
<div><p class="MsoNormal" style="background:#E4E4E4"><b><font size="2" face="Arial"><span style="font-size:10.0pt;font-family:Arial;font-weight:bold">From:</span></font></b><font size="2" face="Arial"><span style="font-size:10.0pt;font-family:Arial"> <a href="mailto:KwangPoo.Chang@rosalindfranklin.edu" title="KwangPoo.Chang@rosalindfranklin.edu">Chang, Kwang-Poo</a> <o:p></o:p></span></font></p>
</div>
<div><p class="MsoNormal"><b><font size="2" face="Arial"><span style="font-size:10.0pt;
font-family:Arial;font-weight:bold">To:</span></font></b><font size="2" face="Arial"><span style="font-size:10.0pt;font-family:Arial"> <a href="mailto:nandlals@hotmail.com" title="nandlals@hotmail.com">Sharman</a> ; <a href="mailto:hgoto@usp.br" title="hgoto@usp.br">Hiro Goto</a> ; <a href="mailto:elfadil_abass@yahoo.com" title="elfadil_abass@yahoo.com">elfadil
abass</a> <o:p></o:p></span></font></p>
</div>
<div><p class="MsoNormal"><b><font size="2" face="Arial"><span style="font-size:10.0pt;
font-family:Arial;font-weight:bold">Cc:</span></font></b><font size="2" face="Arial"><span style="font-size:10.0pt;font-family:Arial"> <a href="mailto:leish-l@lineu.icb.usp.br" title="leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a>
<o:p></o:p></span></font></p>
</div>
<div><p class="MsoNormal"><b><font size="2" face="Arial"><span style="font-size:10.0pt;
font-family:Arial;font-weight:bold">Sent:</span></font></b><font size="2" face="Arial"><span style="font-size:10.0pt;font-family:Arial"> Sunday, May 22,
2011 1:39 AM<o:p></o:p></span></font></p>
</div>
<div><p class="MsoNormal"><b><font size="2" face="Arial"><span style="font-size:10.0pt;
font-family:Arial;font-weight:bold">Subject:</span></font></b><font size="2" face="Arial"><span style="font-size:10.0pt;font-family:Arial"> Re: [Leish-l]
inquiry<o:p></o:p></span></font></p>
</div>
<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 id="idOWAReplyText41530">
<div><p class="MsoNormal"><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><o:p></o:p></p>
</div>
<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><p class="MsoNormal"><font size="2" face="Arial"><span style="font-size:10.0pt;
font-family:Arial">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><o:p></o:p></p>
</div>
<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><p class="MsoNormal"><font size="2" face="Arial"><span style="font-size:10.0pt;
font-family:Arial">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><o:p></o:p></p>
</div>
<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><p class="MsoNormal"><font size="3" face="Arial"><span style="font-size:12.0pt;
font-family:Arial">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><o:p></o:p></p>
</div>
<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><p class="MsoNormal"><font size="2" face="Arial"><span style="font-size:10.0pt;
font-family:Arial">KP</span></font><o:p></o:p></p>
</div>
<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>
<div id="idSignature79278">
<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>
<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">
</span></font></div><p class="MsoNormal" style="margin-bottom:12.0pt"><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">
<a href="mailto:leish-l-bounces@lineu.icb.usp.br">leish-l-bounces@lineu.icb.usp.br</a> 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> <a href="mailto:leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a><br>
<b><span style="font-weight:bold">Subject:</span></b> Re: [Leish-l] inquiry</span></font><o:p></o:p></p>
</div>
</div>
<div><p><font size="2" face="Times New Roman"><span style="font-size:10.0pt">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" <<a href="mailto:hgoto@usp.br">hgoto@usp.br</a>><br>
Sent: Wednesday, May 18, 2011 12:09 AM<br>
To: "elfadil abass" <<a href="mailto:elfadil_abass@yahoo.com">elfadil_abass@yahoo.com</a>><br>
Cc: <<a href="mailto:leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a>><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 <<a href="mailto:elfadil_abass@yahoo.com">elfadil_abass@yahoo.com</a>>:<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 <<a href="mailto:racha@aub.edu.lb">racha@aub.edu.lb</a>><br>
>> To: saad saad <<a href="mailto:saad1426@gmail.com">saad1426@gmail.com</a>>;
"<a href="mailto:leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a>"<br>
>> <<a href="mailto:leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a>><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: <a href="mailto:leish-l-bounces@lineu.icb.usp.br">leish-l-bounces@lineu.icb.usp.br</a><br>
>> [<a href="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: <a href="mailto:leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a><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>
>> <a href="mailto:Leish-l@lineu.icb.usp.br">Leish-l@lineu.icb.usp.br</a><br>
>> <a href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l">http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l</a><br>
>> _______________________________________________<br>
>> Leish-l mailing list<br>
>> <a href="mailto:Leish-l@lineu.icb.usp.br">Leish-l@lineu.icb.usp.br</a><br>
>> <a href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l">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>
> <a href="mailto:Leish-l@lineu.icb.usp.br">Leish-l@lineu.icb.usp.br</a><br>
> <a href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l">http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l</a><br>
><br>
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