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I agree. How about next week. <br>
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<div id="divRplyFwdMsg" dir="ltr"><font face="Calibri, sans-serif" style="font-size:11pt" color="#000000"><b>From:</b> Myotox <myotox-bounces@biophysics.uleth.ca> on behalf of Steele, Harmen <harmen.steele@umconnect.umt.edu><br>
<b>Sent:</b> Tuesday, November 26, 2019 8:38:30 PM<br>
<b>To:</b> Myotoxin-II discussion <myotox@biophysics.uleth.ca><br>
<b>Subject:</b> Re: [Myotox] Myotoxin discussion</font>
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<p class="x_MsoNormal">It sounds like we should schedule a meeting for next week to discuss this as a group.
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<p class="x_MsoNormal"><span style="font-size:12.0pt">Best regards,</span></p>
<p class="x_MsoNormal"><span style="font-size:12.0pt"> </span></p>
<p class="x_MsoNormal"><span style="font-size:12.0pt">Harmen Steele, Ph.D.</span></p>
<p class="x_MsoNormal">Postdoctoral Researcher</p>
<p class="x_MsoNormal"><span style="font-size:12.0pt"><a href="mailto:harmen.steele@umontana.edu"><span style="color:#0563C1">harmen.steele@umontana.edu</span></a></span></p>
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<p class="x_MsoNormal"><span style="font-size:12.0pt">+406 529 9669 - mobile</span></p>
<p class="x_MsoNormal"><span style="font-size:12.0pt"><a href="callto://harmensteele/"><span style="color:#0563C1">Skype Me: harmensteele</span></a> </span></p>
<p class="x_MsoNormal"><span style="font-size:12.0pt">Amateur Radio Call Sign: K9HBS</span></p>
<p class="x_MsoNormal"><span style="font-size:12.0pt">I am a <a href="http://www.umt.edu/umallies/"><span style="color:#0563C1">UM Ally</span></a></span></p>
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<p class="x_MsoNormal"><b><span style="font-size:12.0pt; color:black">From: </span>
</b><span style="font-size:12.0pt; color:black">Myotox <myotox-bounces@biophysics.uleth.ca> on behalf of "Hazendonk, Paul" <paul.hazendonk@uleth.ca><br>
<b>Reply-To: </b>Myotoxin-II discussion <myotox@biophysics.uleth.ca><br>
<b>Date: </b>Tuesday, November 26, 2019 at 6:18 PM<br>
<b>To: </b>Myotoxin-II discussion <myotox@biophysics.uleth.ca><br>
<b>Subject: </b>Re: [Myotox] Myotoxin discussion</span></p>
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<p class="x_MsoNormal" style="margin-bottom:12.0pt"><span style="font-family:"Arial",sans-serif; color:black">Instability is going to be a problem for sure. What can be done? It is entirely possible that we will need to do experiments for several days at the
time.</span></p>
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<p class="x_MsoNormal"><b><span style="color:black">From:</span></b><span style="color:black"> Myotox <myotox-bounces@biophysics.uleth.ca> on behalf of Steele, Harmen <harmen.steele@umconnect.umt.edu><br>
<b>Sent:</b> Tuesday, November 26, 2019 5:26:18 PM<br>
<b>To:</b> Myotoxin-II discussion <myotox@biophysics.uleth.ca><br>
<b>Subject:</b> Re: [Myotox] Myotoxin discussion</span> </p>
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<p class="x_MsoNormal">Mostly it depends on the lipid composition that we want to make. The individual lipid molecules drive their assembly and the size of liposome formed. Normally I make liposomes of 100 nm and try to use them the same day. Beyond a day
and liposomes without special stabilizers have been shown to start to interact and change shape. Once we have a better idea of the lipid composition that we want I can give a better idea of the liposome size we can make.
<br>
<br>
I have not had time today to sit down and research a lipid composition that would make sense to study muscle cells. I will try and work on that tomorrow.<br>
<br>
<br>
Best regards,<br>
<br>
Harmen Steele, Ph.D.<br>
Postdoctoral Researcher<br>
harmen.steele@umontana.edu<br>
<br>
+406 529 9669 - mobile<br>
Skype Me: harmensteele <callto://harmensteele/> <br>
Amateur Radio Call Sign: K9HBS<br>
I am a UM Ally <<a href="http://www.umt.edu/umallies/">http://www.umt.edu/umallies/</a>><br>
<br>
This email was sent using 95% recycled binary bits and 99.9% recycled electrons.<br>
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<br>
On 11/26/19, 3:59 PM, "Myotox on behalf of Borries Demeler" <myotox-bounces@biophysics.uleth.ca on behalf of demeler@gmail.com> wrote:<br>
<br>
<br>
Harmen,<br>
what is the smallest phospholipid liposome (diameter) that you can<br>
produce using your methods, and how homogeneous are they? We can test<br>
for homogeneity with AUC.<br>
<br>
-b.<br>
On Tue, Nov 26, 2019 at 03:53:54PM -0600, Bruno Lomonte wrote:<br>
> Hi Harmen and all,<br>
> <br>
> Dr Carlos Muñoz and his postdoc (at Dept of Physics of UNAM-Cuernavaca,<br>
> Mexico) got interested in the antimicrobial action of the myotoxin, and ran<br>
> these screenings with liposomes using protein that we provided from Costa<br>
> Rica<br>
> <br>
> maybe they would like to work together (?), but on the other hand, it may<br>
> become more complicated<br>
> <br>
> if there is anything else that you would like me to clarify, please do not<br>
> hesitate to let me know<br>
> <br>
> best regards, and thanks for your interest<br>
> <br>
> Bruno<br>
> <br>
> <br>
> ++++<br>
> <br>
> <br>
> <br>
> <br>
> On 26/11/2019 10:09, Steele, Harmen wrote:<br>
> > I can make liposomes. What size do we want to use for the experiments? 100nm? 200nm?<br>
> > <br>
> > I have done some similar experiments to the calcein release experiments discussed. The experiments that I discussed would be some very nice sister experiments to the calcein release. What is your relationship with that lab? It could be a collaborative
experiment?<br>
> > <br>
> > Best regards,<br>
> > Harmen Steele, Ph.D.<br>
> > Postdoctoral Researcher<br>
> > harmen.steele@umontana.edu<br>
> > +406 529 9669 - mobile<br>
> > Skype Me: harmensteele <callto://harmensteele/><br>
> > Amateur Radio Call Sign: K9HBS<br>
> > I am a UM Ally <<a href="http://www.umt.edu/umallies/">http://www.umt.edu/umallies/</a>><br>
> > This email was sent using 95% recycled binary bits and 99.9% recycled electrons.<br>
> > <br>
> > On 11/26/19, 8:49 AM, "Myotox on behalf of Borries Demeler" <myotox-bounces@biophysics.uleth.ca on behalf of demeler@gmail.com> wrote:<br>
> > <br>
> > Thanks Harmen for the detailed explanation on the membranes. Paul/Tony, would<br>
> > you suggest we try liposomes first, or try some other membrane system.<br>
> > Harmen: Do you know how to prepare liposomes of homogenous size? We<br>
> > could verify size homogeneity of liposomes on the AUC.<br>
> > Julian: With your permission, I will add you to our mailing list, OK?<br>
> > -Borries<br>
> > On Mon, Nov 25, 2019 at 11:28:21PM -0600, Bruno Lomonte wrote:<br>
> > > Thank you Harmen and greetings to all,<br>
> > ><br>
> > > just a short note to let you know that regarding *liposomes *there is a<br>
> > > group in Mexico already exploring permeabilization (calcein release) caused<br>
> > > by this myotoxin - they are mostly focusing on ergosterol-containing<br>
> > > liposomes (trying to approach fungal-like bilayers), but I though it is<br>
> > > important that we are all aware of it, just to avoid any potential overlaps<br>
> > ><br>
> > > I hope that this is OK with you, and that there may be different models<br>
> > > which do not overlap with those liposome permeabilization studies<br>
> > ><br>
> > > please allow me to copy this message also to my colleague Julian Fernandez,<br>
> > > with whom we work together in the characterization of myotoxins in Costa<br>
> > > Rica<br>
> > ><br>
> > > kind regards,<br>
> > ><br>
> > > Bruno<br>
> > ><br>
> > ><br>
> > > ++++<br>
> > ><br>
> > ><br>
> > ><br>
> > > On 25/11/2019 22:14, Steele, Harmen wrote:<br>
> > > > The SMA synthetic belts are highly negative. In the electrostatic interactions with cytc we could never tell the difference between lipid and belt interactions. To tell the differences in these interactions we had to us the biological belt,
which are neutral in charge. The SMA nanodiscs are uniform in size and shape when there is a membrane bound protein embedded. Without a membrane bound protein (pure lipid NDs), the nanodiscs are more homogeneous in size and shape.<br>
> > > ><br>
> > > > I have done some work using fluorescence anisotropy and various probes that embed in the lipid bilayers to examine the effect that various metal nanoparticles have on bilayer order. The anisotropy gives us a way to study the degrees of freedom
that a probe has in the lipid bilayer. These degrees of freedom can give us an idea of the order or disorder of a lipid bilayer. By altering the lipid composition of a bilayer we could get an idea of what types of lipids are interacting with myotoxin resulting
in leaky membranes. I was thinking we start with a composition that resembles muscle and then alter the bilayer as we hone in on the specific lipid interactions.<br>
> > > ><br>
> > > > I don't think that nanodiscs would be the best model system to study this interaction. I believe that the belt protein provided a lateral pressure on the lipid bilayer that would inhibit and/or negate any changes a particle might have on the
lipid bilayers packing order. I think that liposomes would be a more appropriate system. Additionally, doing some studies with muscle cells would be appropriate too.<br>
> > > ><br>
> > > ><br>
> > > > Best regards,<br>
> > > > Harmen Steele, Ph.D.<br>
> > > > Postdoctoral Researcher<br>
> > > > harmen.steele@umontana.edu<br>
> > > > +406 529 9669 - mobile<br>
> > > > Skype Me: harmensteele <callto://harmensteele/><br>
> > > > Amateur Radio Call Sign: K9HBS<br>
> > > > I am a UM Ally <<a href="http://www.umt.edu/umallies/">http://www.umt.edu/umallies/</a>><br>
> > > > This email was sent using 95% recycled binary bits and 99.9% recycled electrons.<br>
> > > ><br>
> > > > On 11/22/19, 4:55 PM, "Myotox on behalf of Borries Demeler" <myotox-bounces@biophysics.uleth.ca on behalf of demeler@gmail.com> wrote:<br>
> > > ><br>
> > > > Hello all,<br>
> > > > I am forwarding the information from Bruno to the mailing list so we<br>
> > > > have a record there, including the attached papers so you all have<br>
> > > > access to them.<br>
> > > > In addition: Tony/Mike and Paul: Amy has ~ 100 ul of pure myotoxin-II at<br>
> > > > 21 mg/ml (high concentration) available. It is in phosphate buffer as<br>
> > > > far as I know. We could get that to you. If you need more, I am sure<br>
> > > > Bruno could arrange another shipment.<br>
> > > > I also discussed the project with Harmen. He had a lot of interesting<br>
> > > > ideas with respect on how to prepare nanodisks and membranes, and how<br>
> > > > to check the interaction with Mytoxin-II with the membrane. He has been<br>
> > > > working on a very similar project testing destruction of mitochondrial<br>
> > > > membranes by cytochrome-C, Paul, this is the project we wanted to pursue<br>
> > > > with you. Harmen sent some samples to you guys earlier, not sure if they<br>
> > > > were ever analyzed, but perhaps we could pick this up again as well?<br>
> > > > (and Paul, feel free to add this into the discussion for the CFI, since<br>
> > > > it would be another excellent example for the collaboration between our<br>
> > > > groups and justify the CMP-MAS probe).<br>
> > > > Harmen, I'm going to let you chime in and share your ideas on synthetic<br>
> > > > belts, and various types of lipids that could be used for making<br>
> > > > appropriate nanodiscs. Perhaps you could also find the source for the<br>
> > > > muscle tissue membrane lipids you talked about, as well as some other<br>
> > > > more readily available membrane systems we could try.<br>
> > > > Bruno indicated that most membranes would be affected by the toxicity<br>
> > > > to some degree (what's different about the membranes of the snake's<br>
> > > > toxin-producing glands/toxin-conducting tissues??)<br>
> > > > Thanks, -b.<br>
> > > > ----- Forwarded message from Bruno Lomonte <bruno.lomonte@ucr.ac.cr> -----<br>
> > > > Date: Fri, 22 Nov 2019 12:01:25 -0600<br>
> > > > From: Bruno Lomonte <bruno.lomonte@ucr.ac.cr><br>
> > > > To: "Montina, Tony" <tony.montina@uleth.ca>, "Hazendonk, Paul" <paul.hazendonk@uleth.ca>, "Opyr, Michael"<br>
> > > > <michael.opyr@uleth.ca>, Borries Demeler <demeler@gmail.com><br>
> > > > Subject: Re: Research Collab Meeting<br>
> > > > User-Agent: Mozilla/5.0 (Windows NT 10.0; WOW64; rv:60.0) Gecko/20100101 Thunderbird/60.9.1<br>
> > > > Thanks so much to all for your interest in exploring possibilities to<br>
> > > > study the interaction of the myotoxin II with membranes.<br>
> > > > here attached is the paper in which some phospholipid compositions<br>
> > > > were studied using liposomes, back in 1992<br>
> > > > there was also a follow up with an interesting observation about<br>
> > > > potential auto-acylation of the myotoxin (also attached, 1995)<br>
> > > > codes for the crystal structe of the protein are: 1CLP and 1Y4L<br>
> > > > some info about the active region of the toxins here attached too (2001)<br>
> > > > happy to provide any further info, and of course, protein, if you<br>
> > > > find this project is viable and interesting<br>
> > > > special thanks to Borries for bridging our interests<br>
> > > > all the best,<br>
> > > > Bruno<br>
> > > ><br>
> > > > _______________________________________________<br>
> > > > Myotox mailing list<br>
> > > > Myotox@biophysics.uleth.ca<br>
> > > > <a href="http://demeler7.uleth.ca/mailman/listinfo/myotox">http://demeler7.uleth.ca/mailman/listinfo/myotox</a><br>
> > ><br>
> > > --<br>
> > > Bruno Lomonte, Ph.D.<br>
> > > Instituto Clodomiro Picado<br>
> > > Universidad de Costa Rica<br>
> > > San José, 11501<br>
> > > COSTA RICA<br>
> > ><br>
> > > bruno.lomonte@ucr.ac.cr<br>
> > > tel. +506 2511 7888<br>
> > > cel. +506 8392 0012<br>
> > ><br>
> > > _______________________________________________<br>
> > > Myotox mailing list<br>
> > > Myotox@biophysics.uleth.ca<br>
> > > <a href="http://demeler7.uleth.ca/mailman/listinfo/myotox">http://demeler7.uleth.ca/mailman/listinfo/myotox</a><br>
> > _______________________________________________<br>
> > Myotox mailing list<br>
> > Myotox@biophysics.uleth.ca<br>
> > <a href="http://demeler7.uleth.ca/mailman/listinfo/myotox">http://demeler7.uleth.ca/mailman/listinfo/myotox</a><br>
> > <br>
> > _______________________________________________<br>
> > Myotox mailing list<br>
> > Myotox@biophysics.uleth.ca<br>
> > <a href="http://demeler7.uleth.ca/mailman/listinfo/myotox">http://demeler7.uleth.ca/mailman/listinfo/myotox</a><br>
> <br>
> -- <br>
> Bruno Lomonte, Ph.D.<br>
> Instituto Clodomiro Picado<br>
> Universidad de Costa Rica<br>
> San José, 11501<br>
> COSTA RICA<br>
> <br>
> bruno.lomonte@ucr.ac.cr<br>
> tel. +506 2511 7888<br>
> cel. +506 8392 0012<br>
> <br>
> _______________________________________________<br>
> Myotox mailing list<br>
> Myotox@biophysics.uleth.ca<br>
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