<div dir="ltr"><span class="Apple-style-span" style="border-collapse: collapse; font-family: arial, sans-serif; font-size: 13px; ">Dear KP,<div>Thank you very much for your valuable suggestions and information. I should talk to Neeloo. Regarding the parasites, I can give you the Iist. It is in my thesis. Unfortunately, IICB has lost most of them. Many senior Scientists have retired who were in charge of the job. </div>
<div><br></div><div>Kind regards, </div><div> Madhumita</div></span><br><div class="gmail_quote">On Mon, Jun 6, 2011 at 11:49 PM, Chang, Kwang-Poo <span dir="ltr"><<a href="mailto:KwangPoo.Chang@rosalindfranklin.edu">KwangPoo.Chang@rosalindfranklin.edu</a>></span> wrote:<br>
<blockquote class="gmail_quote" style="margin:0 0 0 .8ex;border-left:1px #ccc solid;padding-left:1ex;">
<div lang="EN-US" link="blue" vlink="blue">
<div>
<ol style="margin-top:0in" start="1" type="1">
<li class="MsoNormal" style="color:navy"><font size="2" color="navy" face="Arial"><span style="font-size:10.0pt;font-family:Arial">Neeloo
singh has been doing genomic sequencing of some Indian isolates. You may
want to coordinate with her.</span></font></li>
<li class="MsoNormal" style="color:navy"><font size="2" color="navy" face="Arial"><span style="font-size:10.0pt;font-family:Arial">Do
you have a list of Dr. Bhaduri’s collection in IICB ? If so, is it
available for checking against what I have ?</span></font></li>
<li class="MsoNormal" style="color:navy"><font size="2" color="navy" face="Arial"><span style="font-size:10.0pt;font-family:Arial">Deep
sequencing of Leishmania genomes is very doable and not terribly
expensive. The bottle-neck is the bioinformatic analyses. There is no user
friendly program for Leish genomics in so far as I know. Neeloo may have
some useful information there.</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"> </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</span></font></p>
<p class="MsoNormal"><font size="2" color="navy" face="Arial"><span style="font-size:10.0pt;font-family:Arial;color:navy"> </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"> Madhumita Manna
[mailto:<a href="mailto:madhumita.manna09@gmail.com" target="_blank">madhumita.manna09@gmail.com</a>] <br>
<b><span style="font-weight:bold">Sent:</span></b> Monday, June 06, 2011 4:45
AM</span></font></p><div><div><font size="2" face="Tahoma"></font></div><div class="h5"><font size="2" face="Tahoma"><br>
<b><span style="font-weight:bold">To:</span></b> Chang, Kwang-Poo<br>
<b><span style="font-weight:bold">Subject:</span></b> Re: [Leish-l] inquiry</font></div></div><p></p>
</div><div><div></div><div class="h5">
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">Dear KP,</span></font></p>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">Thank you very much for your valuable suggestions. I am trying to hit
as many genetic sites are possible and we have a plan to establish a depository
of isolates of KA, PKDL for future works. Prof. Bhaduri tried hard to set up
this kind of bank and at that time we have collected and deposited a good
number of isolates in that Parasite Bank at IICB. </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">For entire genome sequencing, a big group needs to work together and
for that collaboration with other group is solicited.</span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">Kind regards,</span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal" style="margin-bottom:12.0pt"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">Madhumita</span></font></p>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">On Sun, Jun 5, 2011 at 11:05 PM, Chang, Kwang-Poo <<a href="mailto:KwangPoo.Chang@rosalindfranklin.edu" target="_blank">KwangPoo.Chang@rosalindfranklin.edu</a>>
wrote:</span></font></p>
<div>
<div>
<div>
<p class="MsoNormal"><font size="2" color="black" face="Arial"><span style="font-size:10.0pt;font-family:Arial;color:black">Dear Madhumitsa,</span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="2" face="Arial"><span style="font-size:10.0pt;font-family:Arial">You are doing something very important. I re-iterate my
suggestions as follows:</span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="2" face="Arial"><span style="font-size:10.0pt;font-family:Arial">1. Examine Leishmania grown in cultures as well as in the
original samples</span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="2" face="Arial"><span style="font-size:10.0pt;font-family:Arial">2. Evaluate as many genetic sites as possible, preferably
the entire genomes</span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="2" face="Arial"><span style="font-size:10.0pt;font-family:Arial">3. Set up a depository center for Leishmana samples</span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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></p>
</div>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<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>
<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"> Madhumita Manna
[mailto:<a href="mailto:madhumita.manna09@gmail.com" target="_blank">madhumita.manna09@gmail.com</a>]</span></font></p>
</div>
<p class="MsoNormal"><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"> Sun 6/5/2011
7:14 AM<br>
<b><span style="font-weight:bold">To:</span></b> Chang, Kwang-Poo</span></font></p>
<div>
<div>
<p class="MsoNormal"><font size="2" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma"><br>
<b><span style="font-weight:bold">Subject:</span></b> Re: [Leish-l] inquiry</span></font></p>
</div>
</div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<div>
<div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">Dear KP </span></font></p>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">I have collected the isolate in the year 2010 from a patient admitted in
Calcutta National Medical
College and done the
RAPD, RFLP and sequencing in recent past. The publication is underway. The work
is in collaboration with Dr. Syamal Roy, IICB and we are interested in Satluj
river valley isolates as well as we are trying to isolate more clinical
isolates from other parts of India. In this particular case, the transformed
promastigotes were cultured at Bethune
College, Kolkata where we
did not maintain any L.tropica strain or UR6 that time. In 2010, we isolated
five isolates collected from Kolkata of which four patients from
Bihar,one from West Bengal. So, there
was no chance of mixing in this particular case which might result occasionally
due to human errors as you told and also Prof Schnur once cautioned about the
identity of some Indian isolates long back in 1981 when he had screened 84
isolates and he apparently got one L.tropica, if I am not wrong .</span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">I have dealt with UR6 during my Ph.D works in 1993 onwards and have
seen it close to L.tropica [about 72% homology in Jaccard's similarity index
calculated by isozyme and RAPD works]. Recently, by ITS1 RFLP, UR6 is again
found out to be matching with L.tropica. </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">kind regards,</span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">Madhumita</span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal" style="margin-bottom:12.0pt"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">On Sat, Jun 4, 2011 at 11:59 PM, Chang, Kwang-Poo <<a href="mailto:KwangPoo.Chang@rosalindfranklin.edu" target="_blank">KwangPoo.Chang@rosalindfranklin.edu</a>>
wrote:</span></font></p>
<div vlink="blue">
<div>
<p class="MsoNormal"><font size="2" color="navy" face="Arial"><span style="font-size:10.0pt;font-family:Arial;color:navy">It is undoubtedly very important to evaluate the genetics of all
currently existing cultured isolates in India. To be prudent , clones of
these isolates need to be included for examination as well. Over the years, we
have gotten some >300 samples through the kindness of our colleagues in
different forms, ranging from purified DNAs to live/killed parasites to
clinical samples. Sample mix-up does occur in my own lab, regardless of how
meticulous we have tried in handling them. In reality, with no disrespect to my
colleagues, samples received occasionally were not what we expected apparently
due to unintentional errors (most likely of our own in mislabeling). We subsequently
check every sample received by nagt PCR for RFLP or sequencing.</span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<p class="MsoNormal"><font size="2" color="navy" face="Arial"><span style="font-size:10.0pt;font-family:Arial;color:navy">To examine all cultured isolates circulating in India is an
undertaking of great importance that can be best accomplished by a centralized
facility, like ATCC which can guarantee the identity and quality of the
cultures (also for distribution). It will be great to set up a center of such
collection in India
? I am working with Dr. Robert Modestina of ATCC to deposit some of my
collection and will be happy to provide them to any center of Leishmania
collection.</span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<p class="MsoNormal"><font size="2" color="navy" face="Arial"><span style="font-size:10.0pt;font-family:Arial;color:navy">Many years ago, Sarman Singh of AIIMS analyzed a small collection
of India Leishmania cultures he gathered from different Indian colleagues and
brought to my lab, e. g. Ag83, RMRI, DD8, UR6 etc, showing their heterogeneity
by kDNA/nDNA RFLP analyses. That particular Ag83 is identical to 3 primary
cultures of L donovani Bihar isolates, whereas the UR6 is I/II allelic
hetergeneous L tropica in <i><span style="font-style:italic">nagt</span></i>
sequence (see Table [1] of L. odnovani/infantum complex No. 90-93 & Table
[2] L tropica No. 49 in the website: <a href="http://rosalindfranklin.edu/dnn/portals/24/documents/microbiology/sampleList.pdf" target="_blank">http://rosalindfranklin.edu/dnn/portals/24/documents/microbiology/sampleList.pdf</a>).
DNAs were all isolated from uncloned, cultured promastigotes. It is difficult
to trace the origin of the old cultures, which must have been passed around in
different labs over the years. Consequently, I can’t say for sure that the UR6
we examined is identical to yours, for example. This goes with the information
for each isolate provided as well.</span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<p class="MsoNormal"><font size="2" color="navy" face="Arial"><span style="font-size:10.0pt;font-family:Arial;color:navy">Leishmania DNAs in the original sources of clinical, fly and
reservoir samples are more difficult to study, but doable. The potential
problems of using cultured promastigotes can be eliminated by that approach.</span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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 </span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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"> Madhumita Manna [mailto:<a href="mailto:madhumita.manna09@gmail.com" target="_blank">madhumita.manna09@gmail.com</a>]
<br>
<b><span style="font-weight:bold">Sent:</span></b> Friday, June 03, 2011 11:47
PM<br>
<b><span style="font-weight:bold">To:</span></b> Chang, Kwang-Poo</span></font></p>
<div>
<div>
<p class="MsoNormal"><font size="2" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma"><br>
<b><span style="font-weight:bold">Cc:</span></b> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a><br>
<b><span style="font-weight:bold">Subject:</span></b> Re: [Leish-l] inquiry</span></font></p>
</div>
</div>
</div>
<div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">Dear Prof. Chang
and all,</span></font></p>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">Since my Ph.D time
at IICB under Prof. Bhaduri, I am trying to characterize clinical isolates of
KA, PKDL collected mainly from Bihar and very few from West Bengal, by isozyme,
RAPD methods and recently I have come across one isolate having ITS1 sequence
matching with K27. By hsp70 also, the isolate was identified as L.tropica. The
isolate was collected from a patient from Bihar
and he was suffering from KA and sensitive to SAG. This is matching with the
observations made by Sacks in 1995 taking isolates from Bihar.
I have contacted Dr. Sharma for requesting clinical isolates from his lab to
characterize them by isozymes. In order to divulge the population structure of
the causal agents for Indian KA, isolates from different regions of India should be
collected and rigorous typing involving many parameters together, should be
carried forward. </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">Kind regards,</span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">Madhumita </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">on Sat, Jun 4,
2011 at 8:09 AM, Madhumita Manna <<a href="mailto:madhumita.manna09@gmail.com" target="_blank">madhumita.manna09@gmail.com</a>>
wrote:</span></font></p>
<div>
<blockquote style="border:none;border-left:solid #CCCCCC 1.0pt;padding:0in 0in 0in 6.0pt;margin-left:4.8pt;margin-top:5.0pt;margin-right:0in;margin-bottom:5.0pt">
<div>
<p class="MsoNormal" style="margin-bottom:12.0pt"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">----------
Forwarded message ----------<br>
From: <b><span style="font-weight:bold">Chang, Kwang-Poo</span></b> <<a href="mailto:KwangPoo.Chang@rosalindfranklin.edu" target="_blank">KwangPoo.Chang@rosalindfranklin.edu</a>></span></font></p>
</div>
<div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">Date: Fri, Jun 3,
2011 at 11:27 PM<br>
Subject: Re: [Leish-l] inquiry</span></font></p>
</div>
<div>
<div>
<p class="MsoNormal" style="margin-bottom:12.0pt"><font size="3" face="Times New Roman"><span style="font-size:12.0pt">To: "Dr.Sunil
Arora" <<a href="mailto:skarora_in@yahoo.com" target="_blank">skarora_in@yahoo.com</a>>,
<a href="mailto:mmukhtar@tropmedicine.org" target="_blank">mmukhtar@tropmedicine.org</a>,
Sharman <<a href="mailto:nandlals@hotmail.com" target="_blank">nandlals@hotmail.com</a>>,
Petr Volf <<a href="mailto:volf@cesnet.cz" target="_blank">volf@cesnet.cz</a>>,
Hiro Goto <<a href="mailto:hgoto@usp.br" target="_blank">hgoto@usp.br</a>>,
elfadil abass <<a href="mailto:elfadil_abass@yahoo.com" target="_blank">elfadil_abass@yahoo.com</a>>,
<a href="mailto:vishwamohan_katoch@yahoo.co.in" target="_blank">vishwamohan_katoch@yahoo.co.in</a>,
lalit Kant ICMR <<a href="mailto:lalitkant@icmr.org.in" target="_blank">lalitkant@icmr.org.in</a>><br>
Cc: <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a></span></font></p>
<div vlink="blue">
<div>
<p class="MsoNormal"><font size="2" color="navy" face="Arial"><span style="font-size:10.0pt;font-family:Arial;color:navy">Dear Sunil,</span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<p class="MsoNormal"><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></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<ol start="1" type="1">
<li class="MsoNormal" style="color:navy"><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> </li>
</ol>
<p class="MsoNormal" style="margin-left:.25in"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<ol start="2" type="1">
<li class="MsoNormal" style="color:navy"><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 Bihar.
Splenic aspirates from Shyam Sundar’s
VL patients produced exuberant growth as promasigotes in 3N, as I recalled
from my visit to his clinic in the 1980’s (although none of these
promastigotes produces kala-azar in hamsters). This is well-known (correct
me if I should be wrong ?). The genetic sites of these and other Bihar
isolates we examined are different from L infantum strains studied,
including those from China
(Xinjiang, Beijing, Shandong,
Sichuan, Gansu),
Pakistan, Iran, Turkey and elsewhere (see Waki
et al. 2007 Eukaryot. Cell 6, 198). Interestingly, the Bihar isolates is
identical to one Sri Lanka CL isolate (DNA kindly provided by Y Matsumoto,
U Tokyo) in nagt sequence, consistent with published report. Most
interestingly, Shyam Sundar, JC
Dujardin and their colleagues recently reported the isolation of
Leptomonas from ~7% (according to Shyam) of his Bihar
kala-alar samples. This, coupled with the difficulty of in vitro cultures
from clinical samples at least in some cases, underscores the need to
directly examine Leishmania in the original sample sources.</span></font> </li>
</ol>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<ol start="3" type="1">
<li class="MsoNormal" style="color:navy"><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></li>
</ol>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<ol start="4" type="1">
<li class="MsoNormal" style="color:navy"><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></li>
</ol>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<ol start="5" type="1">
<li class="MsoNormal" style="color:navy"><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. Carlos Costa
also told me that he has no difficulty of growing L chagasi from his
patients in Teresina.
He does have a sizable collection of hundreds (or thousands ?) of
isolates, although I was not given the actual rate of successful
isolation.</span></font> </li>
</ol>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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</span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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"> Dr.Sunil Arora [mailto:<a href="mailto:skarora_in@yahoo.com" target="_blank">skarora_in@yahoo.com</a>] <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; <a href="mailto:mmukhtar@tropmedicine.org" target="_blank">mmukhtar@tropmedicine.org</a>;
Sharman; Petr Volf; Hiro Goto; elfadil abass; <a href="mailto:vishwamohan_katoch@yahoo.co.in" target="_blank">vishwamohan_katoch@yahoo.co.in</a>;
lalit Kant ICMR </span></font></p>
<div>
<p class="MsoNormal"><font size="2" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma"><br>
<b><span style="font-weight:bold">Cc:</span></b> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a><br>
<b><span style="font-weight:bold">Subject:</span></b> Re: [Leish-l] inquiry</span></font></p>
</div>
</div>
<p class="MsoNormal"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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">Dear KP and all</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">I have been reading the exchange of
emails regarding characterisation of leishmania isolates from Himachal in India by Dr
Sharma and KP. Actually I have also been associated with that though very
briefly and had a chance to try-n-culture some of isolates sent by Dr
Sharma. The cultures became positive but we failed to establish good passagable
cultures from any of aspirates/biopsy specimen sent by Dr Sharma, although
my lab has been able to establish some good number of primaray cultures from spleen/BM
aspirates of Ld strains in Chandigarh, although almost all the source
patients belonged to Bihar originally for these. So there is definetely
some thing very different about the leishmania strains that are circulating in
that geographical area of India.
we also had a chance to use a specific PCR (that was developed in my lab and
can distinguish between an Ld and any cutaneous strain). we found about 40-50%
of isolates got Ld specific amplification while others didnot show Ld genome.
So I do believe, there is a mix of Ld and some cutaneous strain in that
area which KP is also refering too. It needs to be investigated further... I
know Dr Sharma is on to such effort soon...</span></font></p>
</div>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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">regards</span></font></p>
</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></p>
</div>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal" style="background:white"><em><i><font size="2" color="#00007f" face="Verdana"><span style="font-size:10.0pt;font-family:Verdana;color:#00007F">Dr Sunil K.Arora</span></font></i></em><i><font size="2" color="#00007f" face="Verdana"><span style="font-size:10.0pt;font-family:Verdana;color:#00007F;font-style:italic"><br>
<em><i><font face="Verdana"><span style="font-family:Verdana">Professor </span></font></i></em><br>
<em><i><font face="Verdana"><span style="font-family:Verdana">In Charge HIV
diagnostic and Disease Monitoring Laboratory</span></font></i></em><br>
<em><i><font face="Verdana"><span style="font-family:Verdana">Department of
Immunopathology</span></font></i></em><br>
<em><i><font face="Verdana"><span style="font-family:Verdana">PGIMER,
Chandigarh-160 012</span></font></i></em><br>
<em><i><font face="Verdana"><span style="font-family:Verdana">Ph.:
0091-172-2755192(Off)</span></font></i></em><br>
<em><i><font face="Verdana"><span style="font-family:Verdana">FAX:
0091-172-2744401, 2745078</span></font></i></em><br>
<em><i><font face="Verdana"><span style="font-family:Verdana">Ph:+91-172-2732863
(Res); 9872866087 (cell)</span></font></i></em><br>
<em><i><font face="Verdana"><span style="font-family:Verdana">email: <a href="mailto:skarora_in@yahoo.com" target="_blank">skarora_in@yahoo.com</a>; <a href="mailto:skarorain@gmail.com" target="_blank">skarorain@gmail.com</a></span></font></i></em></span></font></i></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" <<a href="mailto:KwangPoo.Chang@rosalindfranklin.edu" target="_blank">KwangPoo.Chang@rosalindfranklin.edu</a>><br>
<b><span style="font-weight:bold">To:</span></b> <a href="mailto:mmukhtar@tropmedicine.org" target="_blank">mmukhtar@tropmedicine.org</a>;
Sharman <<a href="mailto:nandlals@hotmail.com" target="_blank">nandlals@hotmail.com</a>>;
Petr Volf <<a href="mailto:volf@cesnet.cz" target="_blank">volf@cesnet.cz</a>>;
Hiro Goto <<a href="mailto:hgoto@usp.br" target="_blank">hgoto@usp.br</a>>;
elfadil abass <<a href="mailto:elfadil_abass@yahoo.com" target="_blank">elfadil_abass@yahoo.com</a>>;
<a href="mailto:vishwamohan_katoch@yahoo.co.in" target="_blank">vishwamohan_katoch@yahoo.co.in</a>;
lalit Kant ICMR <<a href="mailto:lalitkant@icmr.org.in" target="_blank">lalitkant@icmr.org.in</a>>
</span></font></p>
<div>
<p class="MsoNormal"><font size="2" color="black" face="Arial"><span style="font-size:10.0pt;font-family:Arial;color:black"><br>
<b><span style="font-weight:bold">Cc:</span></b> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a></span></font></p>
</div>
<p class="MsoNormal"><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">
Friday, June 3, 2011 12:03 AM </span></font></p>
<div>
<p class="MsoNormal"><font size="2" color="black" face="Arial"><span style="font-size:10.0pt;font-family:Arial;color:black"><br>
<b><span style="font-weight:bold">Subject:</span></b> Re: [Leish-l] inquiry</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></p>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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">Thanks again for the info.</span></font></p>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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">Are there any record available for the
number of successful isolations versus the total number attempted for a given
endemic site ?</span></font></p>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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></p>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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"> Maowia Mukhtar [mailto: <a href="mailto:mmukhtar@tropmedicine.org" target="_blank">mmukhtar@tropmedicine.org</a> ] <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" target="_blank">vishwamohan_katoch@yahoo.co.in</a>;
'lalit Kant ICMR' </span></font></p>
<div>
<p class="MsoNormal"><font size="2" color="black" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma;color:black"><br>
<b><span style="font-weight:bold">Cc:</span></b> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a></span></font></p>
</div>
<div>
<p class="MsoNormal"><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></p>
</div>
</div>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</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></p>
</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 Sudan as well
as between the clinical forms. Sudanese Leishmania donovani is known to
be different from the Indian donovani even in their response to antileishmania
drugs and in their reaction using different diagnostic serology tests. </span></font></p>
</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></p>
</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></p>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</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></p>
</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">Institute</span></font></b><b><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D;font-weight:bold"> of Endemic</span></font></b><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></p>
</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">University</span></font></b><b><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D;font-weight:bold"> of Khartoum</span></font></b></p>
</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></p>
</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">Mobile</span></font></b><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></p>
</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></p>
</div>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
</div>
<div>
<div style="border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0in 0in 0in">
<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:<a href="mailto:KwangPoo.Chang@rosalindfranklin.edu" target="_blank">KwangPoo.Chang@rosalindfranklin.edu</a>]
<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" target="_blank">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" target="_blank">leish-l@lineu.icb.usp.br</a>;
Chang, Kwang-Poo </span></font></p>
<div>
<p class="MsoNormal"><font size="2" color="black" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma;color:black"><br>
<b><span style="font-weight:bold">Subject:</span></b> RE: [Leish-l] inquiry</span></font></p>
</div>
</div>
</div>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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">Dear Maowia,</span></font></p>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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">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></p>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<ol start="1" type="1">
<li class="MsoNormal" style="color:navy;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> </li>
</ol>
<div style="margin-left:.25in">
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<ol start="2" type="1">
<li class="MsoNormal" style="color:navy;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>
</li>
</ol>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<ol start="3" type="1">
<li class="MsoNormal" style="color:navy;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></li>
</ol>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<ol start="4" type="1">
<li class="MsoNormal" style="color:navy;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 USA
was probably brought over decades ago by Donald Hyneman and established in
Syrian Golden hamsters by Leslie Stauber and his students. In the 70’s, I
had the good fortune of working with Dennis Dwyer on his S1 or S2 strains
of this origin. As widely reported in the literature, this is an excellent
model of visceral leishmaniasis, producing typical clinical outcome
consistently, including invariable death and splenic LD loads of up to 10<sup>10</sup>
per heavily infected spleen. I recall I had difficult time to
differentiate these splenic amastigotes into promastigotes. I did see some
motile promastigotes. While a small number remains viable, many died and
appeared as ghosts during the subsequent incubation. This has led me to
suspect selection of cultivable clones. In some, but not all of these
cultures, promastigotes eventually became ‘adapted’ to the culture conditions
and grew, producing consistently up to ~40 X 10<sup>6</sup> cells/ml in
4-5 days in successive passages (starting from ~10<sup>6</sup>/ml = ~ 4-5
generation or replication cycles). The stationary phase promastigotes of
these cultures no longer produce good infection of hamsters, irrespective
of the passage numbers (Isolation of ‘metacyclics” to produce good
infection has since been reported in the literature, if I remember
correctly). These promastigotes are more similar to the Mediterranean L
infantum than to the Indian L donovani of the Bihar
region in the sequence of their single-copy protein-coding genes, as
probably already shown by many based on sequencing other genetic sites. </span></font></li>
</ol>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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></p>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </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"> Maowia Mukhtar [mailto: <a href="mailto:mmukhtar@tropmedicine.org" target="_blank">mmukhtar@tropmedicine.org</a> ] <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" target="_blank">vishwamohan_katoch@yahoo.co.in</a>;
'lalit Kant ICMR' </span></font></p>
<div>
<p class="MsoNormal"><font size="2" color="black" face="Tahoma"><span style="font-size:10.0pt;font-family:Tahoma;color:black"><br>
<b><span style="font-weight:bold">Cc:</span></b> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a></span></font></p>
</div>
<p class="MsoNormal"><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> </p>
</div>
</div>
<div>
<div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</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></p>
</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 Sudan ranging
from VL, PKDL, ML to CL. We have isolated L. donovani MON82 from classic
cutaneous ulcers and typed them using both isoenzyme and molecular typing
techniques. Please check our manuscript (Elamin et al. 2008. </span></font><font size="1" color="black" face="Arial"><span style="font-size:9.0pt;font-family:Arial;color:black">Trans R Soc </span></font><font size="1" color="#1f497d" face="Arial"><span style="font-size:9.0pt;font-family:Arial;color:#1F497D">Trop Med Hyg. 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></p>
</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></p>
</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></p>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</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></p>
</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">Institute</span></font></b><b><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D;font-weight:bold"> of Endemic</span></font></b><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></p>
</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">University</span></font></b><b><font size="2" color="#1f497d" face="Calibri"><span style="font-size:11.0pt;font-family:Calibri;color:#1F497D;font-weight:bold"> of Khartoum</span></font></b></p>
</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></p>
</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">Mobile</span></font></b><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></p>
</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></p>
</div>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
<div>
<div style="border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0in 0in 0in">
<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"> <a href="mailto:leish-l-bounces@lineu.icb.usp.br" target="_blank">leish-l-bounces@lineu.icb.usp.br</a>
[mailto:<a href="mailto:leish-l-bounces@lineu.icb.usp.br" target="_blank">leish-l-bounces@lineu.icb.usp.br</a>]
<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" target="_blank">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" target="_blank">leish-l@lineu.icb.usp.br</a><br>
<b><span style="font-weight:bold">Subject:</span></b> Re: [Leish-l] inquiry</span></font></p>
</div>
</div>
</div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</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></p>
</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></p>
</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></p>
</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"> </span></font></p>
</div>
</div>
<div>
<div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</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" title="volf@cesnet.cz" target="_blank">Petr Volf</a> </span></font></p>
</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></p>
</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" title="KwangPoo.Chang@rosalindfranklin.edu" target="_blank">Chang, Kwang-Poo</a>
; <a href="mailto:nandlals@hotmail.com" title="mailto:nandlals@hotmail.com
CTRL + Click to follow link" target="_blank">Sharman</a>
; <a href="mailto:hgoto@usp.br" title="mailto:hgoto@usp.br
CTRL + Click to follow link" target="_blank">Hiro Goto</a>
; <a href="mailto:elfadil_abass@yahoo.com" title="mailto:elfadil_abass@yahoo.com
CTRL + Click to follow link" target="_blank">elfadil
abass</a> </span></font></p>
</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" title="leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a> </span></font></p>
</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></p>
</div>
</div>
</div>
</div>
<div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</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></p>
</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, Turkey ,
cutaneous L. infantum (now it seems that we are dealing with L.donovani/L.
infantum hybrid) grew very poorely if we isolated them from patients. Only 1 of
25 isolations was succesfull. However, the same strain (confirmed by molecular
methods) grew repeatedly and very well if we isolated them from sandflies. It
might be useful for Dr. Sharma try to get isolates from sand flies. It is very
laborious but very useful, in Cukurova we got about dozen of isolates by this
method (all identical).</span></font></p>
</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></p>
</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></p>
</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></p>
</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>
<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></p>
</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" title="KwangPoo.Chang@rosalindfranklin.edu" target="_blank">Chang, Kwang-Poo</a>
</span></font></p>
</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" title="nandlals@hotmail.com" target="_blank">Sharman</a> ; <a href="mailto:hgoto@usp.br" title="hgoto@usp.br" target="_blank">Hiro Goto</a> ; <a href="mailto:elfadil_abass@yahoo.com" title="elfadil_abass@yahoo.com" target="_blank">elfadil abass</a> </span></font></p>
</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">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" title="leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a> </span></font></p>
</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">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></p>
</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">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></p>
</div>
</div>
<div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</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">Dr. NL Sharma has been working on an
important CL endemic area along the Satluj
River valley to the south of
Himalaya in India
(Please correct me should I be wrong for anything I said here and below). I had
the good fortune of visiting the site several years back courtesy of Dr.
Sharma. </span></font></p>
</div>
</div>
<div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</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">I believe Dr. Sharma's finding is
important, since the parasites there are different from the familiar Indian L
donovani in Bihar . The parasites are
refratory to in vitro cultivation. They do differentiate into promastigotes and
may grow a little, but can't really be subcultured to establish
passageable lines. This is very much reminescent of L infantum in the
Mediterranean area. I recall Dr. Sharma has also found rK39+ dogs, if I
remember correctly. If so, Satluj river valley is endemic to
the infantile CL.</span></font></p>
</div>
</div>
<div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</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">Analyses of several batches of basically
clinical CL samples from Dr. Sharma showed evidence of L infantum, but also L
tropica as well as a mixture of the two in one sample. This is the same
picture we have noted for samples from Hatay
, Turkey .</span></font></p>
</div>
</div>
<div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</div>
</div>
<div>
<div>
<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></p>
</div>
</div>
<div>
<div>
<p class="MsoNormal" style="background:white"><font size="3" face="Times New Roman"><span style="font-size:12.0pt"> </span></font></p>
</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">KP</span></font></p>
</div>
</div>
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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"> <a href="mailto:leish-l-bounces@lineu.icb.usp.br" target="_blank">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" target="_blank">leish-l@lineu.icb.usp.br</a><br>
<b><span style="font-weight:bold">Subject:</span></b> Re: [Leish-l] inquiry</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" <<a href="mailto:hgoto@usp.br" target="_blank">hgoto@usp.br</a>><br>
Sent: Wednesday, May 18, 2011 12:09 AM<br>
To: "elfadil abass" <<a href="mailto:elfadil_abass@yahoo.com" target="_blank">elfadil_abass@yahoo.com</a>><br>
Cc: <<a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">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" target="_blank">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>
>> Institute
of Medical Microbiology
and Hospital Hygiene<br>
>> Philipps University Marburg<br>
>> BMFZ / Hans-Meerwein Straße 2<br>
>> D-35033 Marburg , Germany<br>
>> ________________________________________________________________________________<br>
>><br>
>><br>
>><br>
>><br>
>> ________________________________<br>
>> From: Nuha Nuwayri-Salti <<a href="mailto:racha@aub.edu.lb" target="_blank">racha@aub.edu.lb</a>><br>
>> To: saad saad <<a href="mailto:saad1426@gmail.com" target="_blank">saad1426@gmail.com</a>>;
"<a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a>"<br>
>> <<a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">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" target="_blank">leish-l-bounces@lineu.icb.usp.br</a><br>
>> [<a href="mailto:leish-l-bounces@lineu.icb.usp.br" target="_blank">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" target="_blank">leish-l@lineu.icb.usp.br</a><br>
>> Subject: [Leish-l] inquiry<br>
>><br>
>> Dear all<br>
>> Hi. i am Saad from Saudi
Arabia . i have 60 sera from positive case
of<br>
>> CL from south west of the country. I would like to have your<br>
>> recommendations to start a good research line in CL using these sera.<br>
>> Thanks in advance for your help<br>
>> _______________________________________________<br>
>> Leish-l mailing list<br>
>> <a href="mailto:Leish-l@lineu.icb.usp.br" target="_blank">Leish-l@lineu.icb.usp.br</a><br>
>> <a href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l" target="_blank">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" target="_blank">Leish-l@lineu.icb.usp.br</a><br>
>> <a href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l" target="_blank">http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l</a><br>
>><br>
><br>
><br>
><br>
> Profa. Dra. Hiro Goto<br>
> Laboratório de Soroepidemiologia e Imunobiologia<br>
> Instituto de Medicina Tropical de São
Paulo , USP<br>
> Av. Dr. Enéas de Carvalho Aguiar, 470, prédio II, quarto andar<br>
> 05403-000 - São Paulo
, SP<br>
> Tel. +55-11-3061 7023, 3061 7056 ou 3061 7027<br>
> Fax. +55-11-3061-8270<br>
><br>
> _______________________________________________<br>
> Leish-l mailing list<br>
> <a href="mailto:Leish-l@lineu.icb.usp.br" target="_blank">Leish-l@lineu.icb.usp.br</a><br>
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><br>
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