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[Music]

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[Applause]

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thank you very much the second part of

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my talk will be about the experimental

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side of my model the question we want to

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answer here is can we experimentally

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reproduce something like an evolution

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process on a system which does not self

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replicate and as always as mentioned

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before whenever we want something to

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evolve we need to follow a very

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important principle which has to be

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followed otherwise nothing is happening

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we need to apply a constant non

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equilibrium equilibrium we need to have

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a situation where we have a constant non

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equilibrium phase where the complexity

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can rise over time how can we do that

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well the simplest way to do that is to

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take any reaction condition for example

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moisture switch that back and forth

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change that periodically and hereby

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create something like a moving target

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for the process and this moving target

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forces the process to go on in the non

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equilibrium State

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examples are well known dry weight

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cycling and dry wet cycling like I said

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the moisture is changed or the chemical

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potential of water is changed and it's a

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very powerful situation for

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polymerization or even of very efficient

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polymerization doing that I've mentioned

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two papers here of people who are

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present which deal with this efficiency

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of that process now we are looking

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something more exotic we're looking at a

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periodic change of the face state of

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carbon dioxide in a water carbon dioxide

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two-phase system so we have water carbon

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dioxide in the we change the face state

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of carbon dioxide from supercritical

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where it's a very good solvent to

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subcritical where it's a very bad salt

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and so it's actually switching back and

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forth of the solvent quality of carbon

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dioxide okay now where do we expect

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things like that to happen of course

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that's very obvious in case of dry

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weight cycling anywhere on the solids or

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surface of Earth

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however the second one's more difficult

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or the second one we have to look at a

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very specific position within the earth

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crust at about one kilometer of depth

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where the conditions are such

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but little variations of pressure will

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induce this face cycling of the carbon

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dioxide okay and that's exactly the

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environment we want to simulate in the

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laboratory so this is our laboratory

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device it's a high pressure device which

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is computer-controlled we can go up to

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1,000 bars we can vary the temperature

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according to a program very flexible on

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that we have this is the pressure cell

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itself from the outside that's how it

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looks in the inside it's filled with

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water and carbon dioxide so these are

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the two bulk solvents that we have to

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bulk solvents up here then we add a

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mixture of two very primitive

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amphiphiles which is a long-chain fatty

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acid and a long-chain a mine following

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Ashley recommendation by David Deamer

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and we add a set of amino acids which

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are known or which actually expected to

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form under hydrothermal conditions where

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six of them happen to be polar and six

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of them happen to be on the more

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nonpolar side otherwise there's nothing

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else no activation no other chemistry

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involved okay these are the conditions

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120 degrees mainly due to speed up

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things a little bit a relatively high

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temperature then we switch back and

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forth the pressure which allows us to

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undergo this phase transition here the

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phase transition occurs at 72 bars we

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switch between 67 and 77 and the whole

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experiment the source standard with the

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latest experiments is being run for five

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hundred hours which is approximately

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three weeks which means 1500 cycles with

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a period of 20 minutes okay what does

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this phase cycling do well first it does

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very something very similar to what dry

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cycling it also involves condensation

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whenever we bring the carbon dioxide in

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the supercritical phase it suddenly

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takes up water so we decrease the

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chemical potential of water and we

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reinforce the condensation process to

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occur so if we have amino acids here we

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will get more of the condensated

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products when the

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carbon dioxide becomes subcritical again

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then the water is coming back and this

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reinforces hydrolysis so each cycling of

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the pressure means one cycle of

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condensation and hydrolysis okay based

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we have done some experiments with some

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model amino acids to determine the

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kinetics of these processes and based on

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these kinetics we have done some

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calculations just to show how the

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concentration is developed so this is

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the concentration of the amino acids so

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these are the monomers here and on the

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logarithmic scale they hardly change of

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course there is some change but it's

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hardly visible however we have the

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development of the concentration of the

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dimer of the trimer

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of the tetra mer and so on and after a

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day or so the whole thing runs into an

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equilibrium and we end up with having

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decreasing concentrations by an order of

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magnitude for each additional amino acid

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that comes to the chain that's the

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equilibrium situation without any

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activation of course the yield of for

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example the hexapeptide is terribly low

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extremely low yields in this case okay

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but there is something else when we do

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this cycling of the pressure we also

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induce the generation of vesicles reduce

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basically generations one thousand five

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hundred and in the case of our latest

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experiment so we have many generations

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of vesicles which are formed by the

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following mechanism when we are in the

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supercritical state the carbon dioxide

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is saturated with water contains the

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amplifier contains some of the amino

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acids then the supercritical the carbon

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dioxide becomes subcritical so it's a

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bad solvent first water droplets will

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condensate in the mid air in the gas

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phase and the other constituents have

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nothing else to go for nowhere else to

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go then to go in the inside of these

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droplets or on the surface of these

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droplets which the emphathize will do

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when these droplets settle down the

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surface the mono layer on the droplets

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and the mono layer on the surface will

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interact and this will lead to the

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formation

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bicycles and of course these vesicles

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are thermodynamically unstable so after

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a short period of time especially when

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the carbon dioxide become so character

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again they fall apart that means that

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every cycling of the pressure means one

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generation of vesicles okay this is how

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they look like under the microscope when

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in studies some of them are mono lamella

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some of them multilamellar multilamellar

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ones derived from merging droplets in

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the gas phase sizes are around several

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tens of micrometers this would be the

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average size some of them are smaller

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too okay so now we have a pool of

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peptides on one side and we have

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vesicles on the other side now what we

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expect of course is interaction not that

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much of the hydrophilic ones and other

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hand completely hydrophobic ones they

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will have their own things to do but the

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amphiphilic ones some of these peptides

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are amphiphilic and they are capable of

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migrating into the bilayer and when they

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do that they will get protected against

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hydrolysis and therefore they will be

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will get pertained they will accumulate

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over time this accumulation can be

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calculated as well this is the same

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graph as before now I assume that we

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have something like 10% of hexamer which

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is amphiphilic so we start with 10% and

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a few now and if in the calculation now

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it's entered that this amphiphilic

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hexamer is somewhat protected against

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hydrolysis we see a development of the

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concentration and if we look at longer

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periods of time those 500 hours for

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example we see that the concentration of

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this hexamer will keep rising it will

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even get close to the concentration of

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the tetramer and that's exactly what we

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see in the analysis there is an analysis

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done on two experiments actually one is

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without any amphiphilic material so it's

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without vesicles and ones with vesicles

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these are the peptides that we observe

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over times different samples which have

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been taken in case of the absence of the

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amphiphilic material we see that we form

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a relatively large portion of trimer

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smaller quantities but larger varieties

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of tetra mares and hardly any panda

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mares or hexamers in case of the

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experiment with the vesicles it looks

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completely different

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the tetra mares are mostly disappeared

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except for at the end of the experiment

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looks a bit different we don't know why

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but what's really striking is that we

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get a lot of pain tamir's hexa mares and

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also much longer chains which have not

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included in this graph and when looking

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at the longer chains at Paynter mares

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and hexamers we very often find that

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they have allies in a positively charged

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lysine at the head group and more or

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less hydrophobic ones at the other ends

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of the chain and if I label these

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peptides here or the red ones now fall

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into this category then we see that they

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may occur exit occasionally in the first

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experiment but in the second experiment

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they really accumulate so we believe we

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have this accumulation effect we also

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have varied the reaction conditions for

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example in the evolution experiment we

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have after one and a half weeks removed

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the alpha files or thereby vesicles then

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we see a slightly different picture here

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but still we see the accumulation of

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amphiphilic peptides so believe we have

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this integration and this accumulation

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step which is visualized here but we can

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have more when the peptides get longer

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they actually add a they get the

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property of stabilizing or they may get

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the property of stabilizing the vesicles

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especially when they are longer than six

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hundred seven units eight units they may

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stabilize this mythical and when they do

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that and there's a mutual advantage for

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the peptide as well as for the vesicle

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and that means those peptides will get a

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much stronger selectional advantage as

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opposed to the others and it can go even

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further can go to this state which are

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called functional because there we

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developed or the vesicle may develop

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survival strategies may survival

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functions for example when the vesicles

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been formed

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there's osmotic pressure due to the

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concentration gradient between the

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inside and the out

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this osmotic pressure shortens the

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lifetime now if there are channels in

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this concentration gradient can release

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then of course there's an additional

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rate of survival due to that so these

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mechanisms get selected they get

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continuously selected during this 1,500

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generations that we create so that means

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we get additional effects here of course

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we have looked at these peptides here

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especially the longest ones which seem

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to accumulate relatively fast this one

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we were most interested in because it

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accumulated at a relatively high rate

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and we were having this peptide

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synthesized commercially and then we

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added it to a larger portions of these

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vesicles

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and try to find out what the effect

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would be of that peptide on the

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properties of the vesicles we found

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three things we found first that the

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peptide reduces the vesicle size by

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approximately 50% the vesicle sorry the

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peptide increases the vesicle

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permeability by 90% and the peptide

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increases the vesicle stability the

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half-life time is increased by a factor

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of 6 and we interpret that now as

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survival strategies for the vesicle with

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the last issue that's obvious after

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vesicles thermodynamic stable it has

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more chances to survive second one could

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be the effect that an increase of the

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vesicle membrane permeability would

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reduce the osmotic pressure effect and

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would hereby relax the the force which

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could destroy the vesicles and even the

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first one may be a survival strategy

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because smaller vesicles have better

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chances to survive shear or to survive

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bubbling okay so we believe at this

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point we are we actually have functional

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vesicles which actually have developed

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by themselves and at this point of

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course we are looking for more the

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experiments are ongoing especially the

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analyses are ongoing

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they are quite demanding okay this is a

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summary at the end let me briefly

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mention the other persons involved this

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00:13:21,130 --> 00:13:18,110
is Oliver Schmitz who is doing the

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00:13:24,790 --> 00:13:21,140
analyses with his coworkers this is Ivor

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00:13:26,829 --> 00:13:24,800
who is giving us input on the geological

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00:13:29,230 --> 00:13:26,839
side and this is mafia Davila with just

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on maternal leave since half a year

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which gives me myself the chance to work

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on the high-pressure apparatus which is