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</o:shapelayout></xml><![endif]--></head><body lang=NL-BE link=blue vlink=purple><div class=WordSection1><p class=MsoNormal><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Dear K-P, dear all, just to inform you that together with Sanger, we sequenced and analysed the full genome of more than 50 Ldonovani strains (results shortly available) and that we dispose of the needed bio-informatic platform for these diversity analyses. We are extremely interested in increasing the representativity of our L.donovani sample (currently Bihar and Terai).<o:p></o:p></span></p><p class=MsoNormal><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Best regards<o:p></o:p></span></p><p class=MsoNormal><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>JC<o:p></o:p></span></p><p class=MsoNormal><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'><o:p> </o:p></span></p><div><p class=MsoNormal><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Prof. Dr Jean-Claude Dujardin<o:p></o:p></span></p><p class=MsoNormal><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Head of Molecular Parasitology<o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Instituut voor Tropische Geneeskunde<o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Nationalestraat, 155<o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>B-2000 Antwerpen<o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Belgium<o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Tel. 32.3.2476358 or 355<o:p></o:p></span></p><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Fax. 32.3.2476359<o:p></o:p></span></p></div><p class=MsoNormal><span style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'><o:p> </o:p></span></p><div><div style='border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0cm 0cm 0cm'><p class=MsoNormal><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'>From:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'> leish-l-bounces@lineu.icb.usp.br [mailto:leish-l-bounces@lineu.icb.usp.br] <b>On Behalf Of </b>Chang, Kwang-Poo<br><b>Sent:</b> dinsdag 7 juni 2011 14:03<br><b>To:</b> Madhumita Manna; leish-l@lineu.icb.usp.br<br><b>Subject:</b> Re: [Leish-l] inquiry<o:p></o:p></span></p></div></div><p class=MsoNormal><o:p> </o:p></p><div id=idOWAReplyText50965><div><p class=MsoNormal><span style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>I'll be happy to make my collection of Leishmania availalbe to anyone who is interested in setting up a 'permanent' depository of such samples.</span><o:p></o:p></p></div><div><p class=MsoNormal> <o:p></o:p></p></div><div><p class=MsoNormal><span style='font-size:10.0pt;font-family:"Arial","sans-serif"'>KP</span><o:p></o:p></p></div></div><div><p class=MsoNormal><o:p> </o:p></p><div class=MsoNormal align=center style='text-align:center'><hr size=2 width="100%" align=center></div><p class=MsoNormal style='margin-bottom:12.0pt'><b><span style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'>From:</span></b><span style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'> Madhumita Manna [mailto:madhumita.manna09@gmail.com]<br><b>Sent:</b> Tue 6/7/2011 1:29 AM<br><b>To:</b> Chang, Kwang-Poo; leish-l@lineu.icb.usp.br<br><b>Subject:</b> Re: [Leish-l] inquiry</span><o:p></o:p></p></div><div><div><p class=MsoNormal><span class=apple-style-span><span style='font-size:10.0pt;font-family:"Arial","sans-serif"'>Dear KP, </span></span><span class=apple-style-span><span style='font-size:10.0pt;font-family:"Arial","sans-serif"'><o:p></o:p></span></span></p><div><p class=MsoNormal><span style='font-size:10.0pt;font-family:"Arial","sans-serif"'>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. </span><o:p></o:p></p></div><div><p class=MsoNormal><span style='font-size:10.0pt;font-family:"Arial","sans-serif"'><o:p> </o:p></span></p></div><div><p class=MsoNormal><span style='font-size:10.0pt;font-family:"Arial","sans-serif"'>Kind regards, <o:p></o:p></span></p></div><div><p class=MsoNormal><span style='font-size:10.0pt;font-family:"Arial","sans-serif"'> Madhumita<o:p></o:p></span></p></div><p class=MsoNormal><o:p> </o:p></p><div><p class=MsoNormal>On Mon, Jun 6, 2011 at 11:49 PM, Chang, Kwang-Poo <<a href="mailto:KwangPoo.Chang@rosalindfranklin.edu">KwangPoo.Chang@rosalindfranklin.edu</a>> wrote:<o:p></o:p></p><div><div><ol start=1 type=1><li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;mso-list:l0 level1 lfo1'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>Neeloo singh has been doing genomic sequencing of some Indian isolates. You may want to coordinate with her.</span><span lang=EN-US> <o:p></o:p></span></li><li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;mso-list:l0 level1 lfo1'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>Do you have a list of Dr. Bhaduri’s collection in IICB ? If so, is it available for checking against what I have ?</span><span lang=EN-US> <o:p></o:p></span></li><li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;mso-list:l0 level1 lfo1'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>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><span lang=EN-US> <o:p></o:p></span></li></ol><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>KP</span><span lang=EN-US><o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><div><div class=MsoNormal align=center style='text-align:center'><span lang=EN-US><hr size=2 width="100%" align=center></span></div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'>From:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'> Madhumita Manna [mailto:<a href="mailto:madhumita.manna09@gmail.com" target="_blank">madhumita.manna09@gmail.com</a>] <br><b>Sent:</b> Monday, June 06, 2011 4:45 AM</span><span lang=EN-US><o:p></o:p></span></p><div><div><p class=MsoNormal><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'><br><b>To:</b> Chang, Kwang-Poo<br><b>Subject:</b> Re: [Leish-l] inquiry</span><span lang=EN-US><o:p></o:p></span></p></div></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>Dear KP,<o:p></o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>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. <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>For entire genome sequencing, a big group needs to work together and for that collaboration with other group is solicited.<o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>Kind regards,<o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;margin-bottom:12.0pt'><span lang=EN-US>Madhumita<o:p></o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>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:<o:p></o:p></span></p><div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>Dear Madhumitsa,</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>You are doing something very important. I re-iterate my suggestions as follows:</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>1. Examine Leishmania grown in cultures as well as in the original samples</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>2. Evaluate as many genetic sites as possible, preferably the entire genomes</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>3. Set up a depository center for Leishmana samples</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>KP</span><span lang=EN-US><o:p></o:p></span></p></div></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><div class=MsoNormal align=center style='text-align:center'><span lang=EN-US><hr size=2 width="100%" align=center></span></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'>From:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'> Madhumita Manna [mailto:<a href="mailto:madhumita.manna09@gmail.com" target="_blank">madhumita.manna09@gmail.com</a>]</span><span lang=EN-US><o:p></o:p></span></p></div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'>Sent:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'> Sun 6/5/2011 7:14 AM<br><b>To:</b> Chang, Kwang-Poo</span><span lang=EN-US><o:p></o:p></span></p><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'><br><b>Subject:</b> Re: [Leish-l] inquiry</span><span lang=EN-US><o:p></o:p></span></p></div></div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>Dear KP <o:p></o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>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 .<o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>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. <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>kind regards,<o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>Madhumita<o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;margin-bottom:12.0pt'><span lang=EN-US> <o:p></o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>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:<o:p></o:p></span></p><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";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><span lang=EN-US><o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";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><span lang=EN-US><o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";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>nagt</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><span lang=EN-US><o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";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><span lang=EN-US><o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>KP </span><span lang=EN-US><o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><div><div class=MsoNormal align=center style='text-align:center'><span lang=EN-US><hr size=2 width="100%" align=center></span></div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'>From:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'> Madhumita Manna [mailto:<a href="mailto:madhumita.manna09@gmail.com" target="_blank">madhumita.manna09@gmail.com</a>] <br><b>Sent:</b> Friday, June 03, 2011 11:47 PM<br><b>To:</b> Chang, Kwang-Poo</span><span lang=EN-US><o:p></o:p></span></p><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'><br><b>Cc:</b> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a><br><b>Subject:</b> Re: [Leish-l] inquiry</span><span lang=EN-US><o:p></o:p></span></p></div></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>Dear Prof. Chang and all,<o:p></o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>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. <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>Kind regards,<o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>Madhumita <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>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:<o:p></o:p></span></p><div><blockquote style='border:none;border-left:solid #CCCCCC 1.0pt;padding:0cm 0cm 0cm 6.0pt;margin-left:4.8pt;margin-top:5.0pt;margin-right:0cm;margin-bottom:5.0pt'><div><p class=MsoNormal style='mso-margin-top-alt:auto;margin-bottom:12.0pt'><span lang=EN-US> <o:p></o:p></span></p><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>---------- Forwarded message ----------<br>From: <b>Chang, Kwang-Poo</b> <<a href="mailto:KwangPoo.Chang@rosalindfranklin.edu" target="_blank">KwangPoo.Chang@rosalindfranklin.edu</a>><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US>Date: Fri, Jun 3, 2011 at 11:27 PM<br>Subject: Re: [Leish-l] inquiry<o:p></o:p></span></p></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;margin-bottom:12.0pt'><span lang=EN-US>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><o:p></o:p></span></p><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>Dear Sunil,</span><span lang=EN-US><o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";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><span lang=EN-US><o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><ol start=1 type=1><li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;mso-list:l6 level1 lfo2'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>It is important for you and Dr. Sharma to work together, which is very much in line with the spirit of WorldLeish.</span><span lang=EN-US> <o:p></o:p></span></li></ol><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;margin-left:18.0pt'><span lang=EN-US> <o:p></o:p></span></p><ol start=2 type=1><li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;mso-list:l7 level1 lfo3'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>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><span lang=EN-US> <o:p></o:p></span></li></ol><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><ol start=3 type=1><li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;mso-list:l9 level1 lfo4'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>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><span lang=EN-US><o:p></o:p></span></li></ol><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><ol start=4 type=1><li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;mso-list:l8 level1 lfo5'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>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><span lang=EN-US><o:p></o:p></span></li></ol><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><ol start=5 type=1><li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;mso-list:l5 level1 lfo6'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>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><span lang=EN-US> <o:p></o:p></span></li></ol><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>KP</span><span lang=EN-US><o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><div><div class=MsoNormal align=center style='text-align:center'><span lang=EN-US><hr size=2 width="100%" align=center></span></div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'>From:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'> Dr.Sunil Arora [mailto:<a href="mailto:skarora_in@yahoo.com" target="_blank">skarora_in@yahoo.com</a>] <br><b>Sent:</b> Thursday, June 02, 2011 11:55 PM<br><b>To:</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><span lang=EN-US><o:p></o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif"'><br><b>Cc:</b> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a><br><b>Subject:</b> Re: [Leish-l] inquiry</span><span lang=EN-US><o:p></o:p></span></p></div></div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'>Dear KP and all</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Verdana","sans-serif";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><span lang=EN-US><o:p></o:p></span></p></div></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'>regards</span><span lang=EN-US><o:p></o:p></span></p></div></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:black'>sunil</span><span lang=EN-US><o:p></o:p></span></p></div></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><em><span lang=EN-US style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:#00007F'>Dr Sunil K.Arora</span></em><i><span lang=EN-US style='font-size:10.0pt;font-family:"Verdana","sans-serif";color:#00007F'><br><em><span style='font-family:"Verdana","sans-serif"'>Professor </span></em><br><em><span style='font-family:"Verdana","sans-serif"'>In Charge HIV diagnostic and Disease Monitoring Laboratory</span></em><br><em><span style='font-family:"Verdana","sans-serif"'>Department of Immunopathology</span></em><br><em><span style='font-family:"Verdana","sans-serif"'>PGIMER, Chandigarh-160 012</span></em><br><em><span style='font-family:"Verdana","sans-serif"'>Ph.: 0091-172-2755192(Off)</span></em><br><em><span style='font-family:"Verdana","sans-serif"'>FAX: 0091-172-2744401, 2745078</span></em><br><em><span style='font-family:"Verdana","sans-serif"'>Ph:+91-172-2732863 (Res); 9872866087 (cell)</span></em><br><em><span style='font-family:"Verdana","sans-serif"'>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></em></span></i><span lang=EN-US><o:p></o:p></span></p></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;margin-bottom:12.0pt;background:white'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>From:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'> "Chang, Kwang-Poo" <<a href="mailto:KwangPoo.Chang@rosalindfranklin.edu" target="_blank">KwangPoo.Chang@rosalindfranklin.edu</a>><br><b>To:</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><span lang=EN-US><o:p></o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'><br><b>Cc:</b> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a></span><span lang=EN-US><o:p></o:p></span></p></div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>Sent:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'> Friday, June 3, 2011 12:03 AM </span><span lang=EN-US><o:p></o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'><br><b>Subject:</b> Re: [Leish-l] inquiry</span><span lang=EN-US><o:p></o:p></span></p></div><div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>Dear Maowia,</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>Thanks again for the info.</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>Are there any record available for the number of successful isolations versus the total number attempted for a given endemic site ?</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>KP</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'>From:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'> Maowia Mukhtar [mailto: <a href="mailto:mmukhtar@tropmedicine.org" target="_blank">mmukhtar@tropmedicine.org</a> ] <br><b>Sent:</b> Wednesday, June 01, 2011 10:41 PM<br><b>To:</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><span lang=EN-US><o:p></o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'><br><b>Cc:</b> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a></span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'>Subject:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'> RE: [Leish-l] inquiry</span><span lang=EN-US><o:p></o:p></span></p></div></div></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Dear KP,</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";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><span lang=EN-US><o:p></o:p></span></p></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Best regards</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Maowia</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Professor Maowia M. Mukhtar</span></b><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Institute of Endemic Diseases</span></b><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>University of Khartoum</span></b><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>P.O. Box 11463, khartoum, Sudan</span></b><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Mobile; +249912234268</span></b><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Fax: +249183779712</span></b><span lang=EN-US><o:p></o:p></span></p></div></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div></div><div><div style='border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0cm 0cm 0cm'><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'>From:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'> Chang, Kwang-Poo [mailto:<a href="mailto:KwangPoo.Chang@rosalindfranklin.edu" target="_blank">KwangPoo.Chang@rosalindfranklin.edu</a>] <br><b>Sent:</b> 02 June 2011 01:42<br><b>To:</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>Cc:</b> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a>; Chang, Kwang-Poo </span><span lang=EN-US><o:p></o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'><br><b>Subject:</b> RE: [Leish-l] inquiry</span><span lang=EN-US><o:p></o:p></span></p></div></div></div></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>Dear Maowia,</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";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><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><ol start=1 type=1><li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;mso-list:l3 level1 lfo7;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>“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><span lang=EN-US> <o:p></o:p></span></li></ol><div style='margin-left:18.0pt'><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><ol start=2 type=1><li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;mso-list:l1 level1 lfo8;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>“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><span lang=EN-US> <o:p></o:p></span></li></ol><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><ol start=3 type=1><li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;mso-list:l2 level1 lfo9;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>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><span lang=EN-US><o:p></o:p></span></li></ol><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><ol start=4 type=1><li class=MsoNormal style='color:navy;mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;mso-list:l4 level1 lfo10;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif"'>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><span lang=EN-US><o:p></o:p></span></li></ol><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:navy'>KP</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'>From:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'> Maowia Mukhtar [mailto: <a href="mailto:mmukhtar@tropmedicine.org" target="_blank">mmukhtar@tropmedicine.org</a> ] <br><b>Sent:</b> Tuesday, May 31, 2011 2:30 PM<br><b>To:</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><span lang=EN-US><o:p></o:p></span></p><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'><br><b>Cc:</b> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a></span><span lang=EN-US><o:p></o:p></span></p></div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'>Subject:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'> RE: [Leish-l] inquiry</span><span lang=EN-US> <o:p></o:p></span></p></div></div><div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Dear Colleagues,</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";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><span lang=EN-US style='font-size:9.0pt;font-family:"Arial","sans-serif";color:black'>Trans R Soc </span><span lang=EN-US style='font-size:9.0pt;font-family:"Arial","sans-serif";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><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:9.0pt;font-family:"Arial","sans-serif";color:#1F497D'>Best regards</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:9.0pt;font-family:"Arial","sans-serif";color:#1F497D'>Maowia</span><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Professor Maowia M. Mukhtar</span></b><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Institute of Endemic Diseases</span></b><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>University of Khartoum</span></b><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>P.O. Box 11463, khartoum, Sudan</span></b><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Mobile; +249912234268</span></b><span lang=EN-US><o:p></o:p></span></p></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D'>Fax: +249183779712</span></b><span lang=EN-US><o:p></o:p></span></p></div></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><div style='border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0cm 0cm 0cm'><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'>From:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";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>On Behalf Of </b>Sharman<br><b>Sent:</b> 31 May 2011 18:13<br><b>To:</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>Cc:</b> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a><br><b>Subject:</b> Re: [Leish-l] inquiry</span><span lang=EN-US><o:p></o:p></span></p></div></div></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-family:"Calibri","sans-serif";color:black'>Hi Petr,</span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-family:"Calibri","sans-serif";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><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-family:"Calibri","sans-serif";color:black'>This particular focus seems to be a complex one.</span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-family:"Calibri","sans-serif";color:black'>NL Sharma</span><span lang=EN-US style='color:black'> </span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div></div><div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:whitesmoke'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'>From:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'> <a href="mailto:volf@cesnet.cz" target="_blank" title="volf@cesnet.cz">Petr Volf</a> </span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:whitesmoke'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'>Sent:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'> Tuesday, May 31, 2011 12:36 PM</span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:whitesmoke'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'>To:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'> <a href="mailto:KwangPoo.Chang@rosalindfranklin.edu" target="_blank" title="KwangPoo.Chang@rosalindfranklin.edu">Chang, Kwang-Poo</a> ; <a href="mailto:nandlals@hotmail.com" target="_blank" title="mailto:nandlals@hotmail.com CTRL + Click to follow link">Sharman</a> ; <a href="mailto:hgoto@usp.br" target="_blank" title="mailto:hgoto@usp.br CTRL + Click to follow link">Hiro Goto</a> ; <a href="mailto:elfadil_abass@yahoo.com" target="_blank" title="mailto:elfadil_abass@yahoo.com CTRL + Click to follow link">elfadil abass</a> </span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:whitesmoke'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'>Cc:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank" title="leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a> </span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:whitesmoke'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'>Subject:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'> Re: [Leish-l] inquiry</span><span lang=EN-US><o:p></o:p></span></p></div></div></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>Hi K.P.,</span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";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><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>Patients are rK39 negative, see attached paper.</span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>Best wishes</span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>Petr</span><span lang=EN-US><o:p></o:p></span></p></div></div><blockquote style='border:none;border-left:solid black 1.5pt;padding:0cm 0cm 0cm 4.0pt;margin-left:3.75pt;margin-top:5.0pt;margin-right:0cm;margin-bottom:5.0pt'><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>----- Original Message ----- </span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:#E4E4E4'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>From:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'> <a href="mailto:KwangPoo.Chang@rosalindfranklin.edu" target="_blank" title="KwangPoo.Chang@rosalindfranklin.edu">Chang, Kwang-Poo</a> </span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>To:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'> <a href="mailto:nandlals@hotmail.com" target="_blank" title="nandlals@hotmail.com">Sharman</a> ; <a href="mailto:hgoto@usp.br" target="_blank" title="hgoto@usp.br">Hiro Goto</a> ; <a href="mailto:elfadil_abass@yahoo.com" target="_blank" title="elfadil_abass@yahoo.com">elfadil abass</a> </span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>Cc:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank" title="leish-l@lineu.icb.usp.br">leish-l@lineu.icb.usp.br</a> </span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>Sent:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'> Sunday, May 22, 2011 1:39 AM</span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>Subject:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'> Re: [Leish-l] inquiry</span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div></div><div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";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><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";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><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";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><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-family:"Arial","sans-serif";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><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='font-size:10.0pt;font-family:"Arial","sans-serif";color:black'>KP</span><span lang=EN-US><o:p></o:p></span></p></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div></div></div><div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US> <o:p></o:p></span></p></div></div><div><div style='margin-bottom:12.0pt'><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black'>From:</span></b><span lang=EN-US style='font-size:10.0pt;font-family:"Tahoma","sans-serif";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>Sent:</b> Fri 5/20/2011 11:31 PM<br><b>To:</b> Hiro Goto; elfadil abass<br><b>Cc:</b> <a href="mailto:leish-l@lineu.icb.usp.br" target="_blank">leish-l@lineu.icb.usp.br</a><br><b>Subject:</b> Re: [Leish-l] inquiry</span><span lang=EN-US><o:p></o:p></span></p></div></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US 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>> <a href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l" target="_blank">http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l</a><br>><br>_______________________________________________<br>Leish-l mailing list<br><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></span><span lang=EN-US><o:p></o:p></span></p></div></div><div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto;background:white'><span lang=EN-US style='color:black'>_______________________________________________<br>Leish-l mailing list<br><a href="mailto:Leish-l@lineu.icb.usp.br" target="_blank">Leish-l@lineu.icb.usp.br</a><br><a href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l" target="_blank">http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l</a></span><span lang=EN-US><o:p></o:p></span></p></div></blockquote></div></div></div></div><div><div><p class=MsoNormal style='mso-margin-top-alt:auto;margin-bottom:12.0pt;background:white'><span lang=EN-US style='color:black'><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></span><span lang=EN-US><o:p></o:p></span></p></div></div></div></div><p class=MsoNormal style='mso-margin-top-alt:auto;margin-bottom:12.0pt'><span lang=EN-US><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><o:p></o:p></span></p></div></div></div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div></blockquote></div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div></div></div></div></div></div></div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div></div></div></div></div></div></div><p class=MsoNormal style='mso-margin-top-alt:auto;mso-margin-bottom-alt:auto'><span lang=EN-US> <o:p></o:p></span></p></div></div></div></div></div></div></div><p class=MsoNormal><o:p> </o:p></p></div></div></div>
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