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Open-Source Technique for GM Crops
a_d_white writes "The Biological Innovation for Open Society has developed TransBacter, a new technique for creating genetically modified crops, which is being released as a BioForge project. Their license allows anyone to use and improve the technique as long as improvements are shared with everyone, à la open source. Other techniques for creating genetically modified crops rely on Agrobacterium, but this new method allows using bacteria outside this genus. The New York Times and Wired cover the story. The founding of BIOS was mentioned previously. Although the Nature paper is available from the BIOS website, with their emphasis on the free sharing of ideas it's rather ironic that the technique was not reported in an open-access journal."
Imagine my disappointment when I misread the title and thought it was some kind of hack for GM cars...
This is nice to see. Information, free for all. In this casr especially, since it helps all of us.
I wonder how many other things would benefit the 'end user' if things were opened. Auto safety for instance.
Pretty Pictures!
I understand BioForge is a place for scientists to collaborate but is it also a place for funding? Did the scientists who put together this article do so with funds from a University or (less likely) a corporation?
If more of these papers are to come out, and I hope they do, the proper funding channels should be lined up since those who fund a research project tend to be very possessive about the results.
-Teiresias
More precisely, "à la the GPL". I know everyone here has "GM plants", Monsanto, terminator seeds and the RIAA muddled together into a single ball of confusion but it's not like public domain vectors haven't been available for, what, 20 years?
At any rate, it's a nice piece of work. The submitter can sneer at them for their choice of journal, but I'd take the Nature paper if I were them.
What I'm listening to now on Pandora...
Their license allows anyone to use and improve the technique as long as improvements are shared with everyone, à la open source.
This is foolish. They should have released it under a free license for anyone except those who deny the same right to use their bio-patents. Otherwise certain scums are able to use this technique while not being forced to change their behaviour, hurting the industry, hurting the farmers, hurting the scientific progress, with no consequences. A perfect license should be useful for cross-licensing with proprietary patents portfolios but sadly this one while being certainly great in spirit is just too weak in its current form to achieve this goal. In the real world of patent sharks we need to fight a little bit harder.
Sincerely,
Pan Tarhei Hosé, PhD.
"Homo sum et cogito ergo odi profanum vulgus et libido."
I've already started. You should see my 5 assed garbanzo beans.
I don't know about every one else, but I've been using Open Source and BIOS technologies together for years now.
i just put in
Still can't do it in my kitche^H^H home lab.
It's still easier to soak seeds in a mutagenic formula, plant them and look for interesting traits later, then clone and reproduce.
GM crops have tremendous potential in regions such as Africa, where also, unfortunately, the governments are too afraid to use GM strains because they risk their agricultural exports with the hysterically-anti-GM nations (because of the fear of cross-polination).
These developing countries can't even compete fairly with unmodified crops because of the unfair subsidies Western governments give their own farmers. Imagine that--taxing your highly advanced industrial complex and then using the money to artificially lower the prices of your products in one of the only markets that people of impoverished nations can compete in!
How long is the developing world going to suffer because technological nations remain sentimental over their own agriculture?
When things get complex, multiply by the complex conjugate.
I remember that a farmer was successfully sued for having GM crops on his farm which were patented by a corporation. It turns out he didn't purposefully plant that strain of crop; wind currents allowed the GM strain to migrate to his farm, where it then began establishing a foothold amongst the farmer's normal crops.
Norman Borlaug inroduced a hybrid wheat strain to India in 1963. It doubled the wheat's yield per acre, leading to a net increase 20 million tons per year from 12 million tons. So obviously, crop strains have a great impact upon the population.
Seems like a way to introduce a harmful gm product as a weapon to destroy a nations food supply. By providing this information so readily it may make the job much easier. Especially as improvements to the techniques are made.
Tomacco will finally become reality
Sorry guys. My modified corn crop not only causes cancer in 90% of all people it also kinda crossbred with the native corn in most of the southwest... so... uh... Sorry Guys.
Just a boy doing unproffesional IT work that's way above his head.
When monsanto crops breed with your GPL crops, they have to release the genetic code or they are in violation of the liscense?
After all Cocaine wants to be free too.
No, you're confusing two things.
Selecting individual plants or animals and breeding strains in the hope of exagerrating desirable traits (resistance to disease, early ripening of fruit, etc). is one thing.
This can only happen within a single species, so far as I know. I might be wrong about this. It happens.
If you manage to get a hybrid of two species, the offspring are sterile, so the strain acnnot ontinue beyond a first generation fo offspring (cf. mules).
What is meant by GM, is taking genetic information from one species and inserting it into the genome of another species. This crossing of the species barrier cannot normaly happen, and certainly has not been used by farmers "for centuries".
Now, while it may be laudable to develop a strain of rapeseed that is resistance to a particular disease by inserting a gene from a bacterium, what happens if pollen from a field full of this rapeseed is taken up by bees and some of this is eaten by another bacterium.
This is what the European Commission is wary of. Monsanto et.el. are pushing for short term profits by being first-to-market. Let's face it, the directors are put there to serve shareholders' interests. "Long term" investment for many of those shareholders is maybe ten years.
The commissionars in Brussels are nominated by career politicians and technocrats, whose short term goals are mainly fiscal but whose long term goals are to return to power over again, in alternating periods of government. Now, we're looking at three to five cycles of five to seven years...
The consumer is torn between the desire for ultra-cheap food right now, this instant, and wanting his childrens and maybe unborn grandchildren to be born with the right number of fingers, toes, eyes and ears.
Beef>
Does biodiversity mean a thing to any of you? Having one strain of GM corn dominate all of an an area's crops might be awesome when it comes to raising productivity levels and immunity to pesticides, but when an unforseen disease starts to affect the plants (which can happen a lot) they would be completely wiped out, because they are all the same. Nature does it better, lets not fuck around with it.
Introducing GM plants to an area can be compared to introducing alien species to a place where they do not belong. There is no possible way to forsee all the negative impacts that could arise. Check out all the problems Australia has with feral animals, for instance. here
http://www.thelung.org
Talk about viral marketing.
--
make install -not war
Personally, I would say that most of the government is as open source as Linux. In other words, the information is there if you bother to look for it and are willing to go through the trouble of interpretting it. The only part that really isn't open is the closed-door meetings and the like. Aren't there any black-box modules in Linux where you only know something goes in and something else goes out? Anyhow, I think the issue is less whether the information is available as it is the complexity of the system. The average politician is unlikely to be able to look at the sourcecode to Linux and understand how it works. The average programmer is unlikely to be able to look at the internal workings of the government and understand how it works.
This sig has absolutely no significance and serves only to take up screen space and waste the time of the reader.
I wrote this the other day in response to hearing about a professor who is all bent out of shape because some corn variety may be gone forever.
It also touches on some surrounding GMO issues, but it really sums up my position on the matter.
---------
As far as GM crops go, there is no stopping scientific progress.
Instead, we should be dealing with how we are going to deal with
possible consequences. If that professor had instead made a research
project out of preserving the DNA of "irreplaceable native Mexican corn
varieties" they would no longer be irreplaceable.
It is my understanding that there is somewhat of a question in
the law right now about GM crops appearing in other fields, and then the
patent owners then billing the field owner for its use... but that is a
problem with our retarded patent system and the law. Besides, I don't
expect the GM companies are going to kick little old ladies off their
maize fields when they don't pay their GM licensing fee... however, it
is not unimaginable in my mind, so if I read about that I would be
pissed. All this does now is give the people a more robust/pest
resistant/productive corn plant.
Yes there definitely are unforeseen consequences that are going to bite
us in the ass, but man will, nay, MUST conquer genetic engineering. Yes
they should be more careful, but they are not, and it might save all our
lives someday. The lives of countless others, the elimination of
suffering from the failures of our "evolved over time to be just
slightly better than the last blunder of design that caused the previous
in our line of species to disappear into time. I know I am being
extreme and this is all very far off, but the more people screwing with
super science the better.
I don't think the analogy with open source software is quite right. After all, with OSS I have a choice - I download the software, .configure, make install and I have it. Then if it I choose I can delete it.
With this stuff I can still choose whether I want to plant it or eat it, but I cannot choose whether my neighbor's GM'd tomatoes pollinate my tomatoes. I won't find out until I plant the resulting seeds next summer and WHOA! My tomatoes have deformed frog legs on them, but geez, they grow like the dickens in my cat's litter box!
I'll leave them on the front porch - help yourself.
PLoS Biology and the other PLoS journals are good, but even though they're open access, they're not repsected the way Nature is. In addition, Nature is a general subject journal, and has a huge readership - more akin to a magazine than a journal. Putting their article in Nature, while reserving the power to distribute it freely on their site, was probably the best way to have impact. And generating news is something that Robert Jefferson is pretty good at.
"In order to make an apple pie from scratch, you must first create the universe." -- Carl Sagan, Cosmos
-- If you manage to get a hybrid of two species, the offspring are sterile, so the strain acnnot ontinue beyond a first generation fo offspring (cf. mules).
Well, not totally... lateral gene transfer (transfer of genes from one speicies to another) has been hapening for millenia - bacteria do it, yeasts do it, and viruses allow higher organisms to do it.
Therefore, anyone who has been making cheese, alcohol, or any fermented food has been engaging in GM for a long time.
"In order to make an apple pie from scratch, you must first create the universe." -- Carl Sagan, Cosmos
Until then I mix my own drinks and use organic celery.
If you are ok with GM stuff then you should also be fine with the intro of non-native species. Why would one be ok and the other not ok? The same fundamental issue exists with both: the unintended consequences of introducing a species alien to this environment are unknown and predictions of "it'll be fine" are often wrong. Go talk to the Australians about introduced species and see if they agree that the human mind is up to the task, last I heard the problems were NOT solved. And yes, if it is genetically modified surely it is a new species!?
90% of the wealth is in 2% of the pockets. Bummer to be in the majority.
This is incorrect. Genes hop across species lines all the time. Microorganisms routinely swap, inject and steal genes on an on going basis even across such divisions as eukaryotes vs prokaryotes. Viruses move genes between multicellular species routinely.
It has always amused me that people fear GM when for the last 100 years the standard breeding method for food crops has been to force mutate them with radiation and mutagenic chemicals. Such practices mutate thousands of unknown mutated genes for every beneficial gene they produce. Nobody ever checked if if 1/10 or 1 percent of the general population was allergic to a protein in a mutated food plant.
At least with GM, we know what we changed and where and when we changed it. With forced mutation and natural gene swapping we have no idea.
There is a whole world of difference between introducing an organism that is genetically modified (=same species + new feature) and a completely new species into an environment.
Of course, it all depends on what this new feature is, but in my opinion, 99% of the modifications we wish to make to a specific crop are beneficial only to us and not to the crop itself (read: its survival in the wild).
For instance, consider a tomato plant that has been modified to grow tomatoes that are twice as big and that can be preserved twice as long as regular tomatoes. While this is obviously beneficial to the farmer and the consumer, it will seriously hinder the survival of this tomato variety in the willd: regular tomato plants will spend less energy on producing fruit and will be able to release their seeds much sooner (because the fruit spoils faster) than the fancy GM variety.
What about liability issues? When it was discovered that asbestos caused cancer, the company who manufactured it was liable. Who's liable if something goes wrong with open-source GM crops?
www.empiresofsteel.com
I don't have the link on me, but there was a recent report that the "Interim" Iraqi government made it illegal for Iraqi farmers to keep seeds. Not only are they going to be forced to only get GM crops from U.S. agribusinesses (the ones that donated to Bush, of course), but they are not even going to be allowed to replant.
I'm expecting this to happen to Afghanistan as well. They are thinking ahead to control of food and water in the third world, rather than just money. Iraq is important for its water supply as well as its oil.
>>"ad space available -- low rates!!!"
While this is obviously beneficial to the farmer and the consumer, it will seriously hinder the survival of this tomato variety in the willd: regular tomato plants will spend less energy on producing fruit and will be able to release their seeds much sooner (because the fruit spoils faster) than the fancy GM variety.
The tomato plants don't have to be able to reproduce themselves in order to spread their genetic code -- all they need to do is release pollen.
And studies have shown that cross-pollination with non-GMO crops can happen, but that often the changes placed in the DNA are dominant traits, meaning that they will be passed down via the pollen.
This has caused a nightmare in the organic farming community, because it's now possible that GMO strains can contaminate their crops, removing their organic status. And once they're contaminated, it's not easy to decontaminate them.
Karma: Chevy Kavalierma.
Isn't science supposed to be about open exhange of information?
Aren't most of the open source licenses based on the open exchange of information that is a major component of research? The BSD license is a good example of it, you can use the code as long as you attribute it correctly. GPL is just placing safe guards so that the information cannot be 'locked up'.
It looks like the wheel has come full circle.
putting the 'B' in LGBTQ+
I don't really think it's ironic - scientific journals are about prestige, and the impact factor of Nature is very high - over 30. I don't think any of the open access journals even have impact factors yet.
If the purpose of the announcement is to reach as many as the biological sciences community as possible, you want to put the announcement in the journal that most of them read.
Yes, there are women on Slashdot. Deal with it.
How about:
3) Move cautiously in a new technology, and always for the right reasons.
From what I've learned so far in my life, preventing a problem is generally cheaper than solving it once it's happened.
We already have indicators that GM is problematic: some species of insect have died eating GM crops; there has been cross-pollination with other crops, sometimes with unexpected results; GM crops have in some cases shown to be of lower quality than normal crops.
With the prolific use of Roundup, there are now Roundup resistant weeds. What will the next step be?
Keep in mind, right now there is no agency regulating or overseeing GM crops. None. Roundup-Ready soybeans are classified, not as a food, but as a pesticide.
Until there is a regulatory agency that can adequately monitor the effects of GM crops and their safety, and until there is labelling of GM crops (which a vast majority of the public supports), I think opposing GM is a good idea in general because it's largely controlled by large corporations.
I believe that free markets work for certain things, but for others they are less than ideal. For example, most of the public schools that are/were run by private companies really suck -- they're using old textbooks and lousy teachers, all to keep up their bottom line.
There are some things in life that shouldn't be profitable enterprises. Monitoring food safety is one. I think that we shouldn't be putting our faith in Monsanto that they're looking out for our food safety.
I think the idea of open-source GM is very appealing. As long as agrobusiness controls GM science, I will be in the opposition group, because I don't believe that these corporations are looking out for our safety interests. Unless they're basically a fiscally irresponsible company, then they're spending the absolute minimum required (legally and proactively to avoid lawsuits) in order to ensure that people won't get really sick from, say, eating a burger made out of GM soy.
As long as agribusiness controls seed, it controls farmers and, ultimately, the food supply. If seed is non-GM or open source GM, then the farmers control the seed and the supply. But if GM remains closed, and there are enough mergers so that there are only two giants left, then for at least a year or two, they could completely control our food supply (until, hopefully, another company was able to rise up to compete against them).
So, although I'm nervous about the potential health and ecological risks of GM food, I'm terrified about Monstanto or an equivalent taking control of the food supply. Imagine if we had a Microsoft of food. That would suck.
That's why, although I'm nervous about GMOs in general, this article is a good sign.
Karma: Chevy Kavalierma.
In free software, we have a long tradition of actors who are half-in, half-out of our community. We benefit from their involvement in some projects, where they are equal participants, even as we may disapprove of their other activities. It gives them the possibility to contribute without making an all-or-nothing committment. In practice, it works out well. IBM, like Monsanto, has been traditionally evil, but they're improving thanks in large part to our community's willingness to work with them. Who knows, maybe we can bring Monsanto around as well. :-)
The evaluation of an action as 'practical' . . . depends on what it is that one wishes to practice.
This is not the "open source" way of doing things.
Open source just requires you release your source code. It does NOT require anyone else release their modifications.
This is the GNU model, forcing others to release their changes.
Pay more attention!
Not so true for plants. Often the diploid offspring are infertile, but conversion to tetraploid form can restore fertility. (This is true for lilium species, at least. For mammals(at least), getting converted to tetraploid form is a bad idea.)
In addition, plant tissue culture makes the issue of fertility somewhat less of an issue, again depending on the plant. Much of the tree-borne fruit that you see in any store (apples, oranges, peaches, I think bananas), was propagated asexually (grafted onto root stock).
The scale of "conventional" techniques for improving species (e.g., plant 10 acres of pink lilies, keep the 100 best stems, crossbreed, repeat for 10 generations) is sufficiently large that I would not bet too much money that accidental gene transfer/modification (by viruses/bacteria/background radiation) isn't occurring anyway. I don't think anyone ever did any formal safety tests on the first navel orange; they saw that it was seedless and tasty enough, and propagated it all over the place.
Agrobacterium is compress. It's basically the only game in town for gene transfer into plants, which (regardless of your views on GM crops) is critical to agroscience. But it's patented. These guys found an alternative, and are bringing a fundamental technique to the free world. It opens up whole new possibilities, and I bet a lot of scientists are going to choose to be part of the free world in coming years. Let's hope the effect turns out to be as revolutionary as GNU.
The evaluation of an action as 'practical' . . . depends on what it is that one wishes to practice.
Such practices mutate thousands of unknown mutated genes for every beneficial gene they produce. Nobody ever checked if if 1/10 or 1 percent of the general population was allergic to a protein in a mutated food plant.
You're missing the point on the concern about allergies.
It's not that a new allergen might be created. Nearly ANYTHING can be allergenic - either directly or in complex with something else in the body. New compounds are being "invented" by mutation all the time. Forcing some mutations into a few plants in a lab is not even a drop in the bucket.
The concern is that there are a lot of compounds that are allergenic AND PERVASIVE in certain food plants. When someone becomes allergic to them, essentially the only effective treatment is to avoid them. (As of the last time I checked, desensitization does NOT work on the branch of the immune system that mediates food allergies.)
But these protiens are often useful: Some of them are the biological warfare agents the crop plants use against the parisites they are resistant to. Copying them into other species of crop plants can confer the resistance. Very useful.
But if you copy, say, a corn allergen to wheat, and sell the modified wheat AS wheat, how is someone who is allergic to it supposed to avoid it. Look at the label: It says "wheat". No corn ingredients (not even the dozen or so that don't sound like corn but are). Ought to be safe, right?
And with engineered wheat containing this corn allergen (and probably several others) immune to a range of pests (so no expensive pesticides are needed to keep its yeilds up), it will soon displace the non-engineered corn from commercial farming. So people with corn allergies won't be able to eat wheat, either.
Heck, how can they even FIGURE OUT that it's a CORN allergy? Scratch tests don't work. You need to do an elimination diet. Where do they start? Just meat? (What if the cattle were corn-fed? Many plant protiens appear in the meat in enough concentration to affect the taste and smell, which is no more sensitive than the immune system.)
Repeat with transferring wheat allergens into corn. Then play three-way musical genes with potatoes. Then add more plants to the mix.
Eventually, if you're allergic to ANY crop plant you're allergic to ALL crop plants. Then what do you eat?
Or are you trying to breed out people who get food allergies? (I suppose that's one way to reduce the population to the handfull that could be fed without the use of pesticides, fertilizers, OR gene-engineered crops.)
Bantam Dominique roosters crow a four-note song. Once you've heard it as "Happy BIRTHday" you can't NOT hear it that way
GM products DO have a few down sides, some of which you mentioned.
They do tend to incourage monoculture farming. However this is nothing new. Farmers have been trying to monoculture for the last 10,ooo years. In the last 50-100 years they have gotten to the point of true monocultures. But just like any other plant they will only thrive in the enviroment they were designed for (by nature or man). If farmers see that they are not doing as well with GM crops THEY WILL STOP USING THEM.
GM foods can be health hazards. Especially if you mix organisms that tend to trigger alergic reactions like peanuts. It could be dangerous for people with sever allergies to buy carrots (or any other food) because it has peanut products in it. While this danger is real it is also avoidable. Just like so many prepared foods already have warning labels for food alergies, GM food will probably need labels.
But here are a few errors in you assumptions:
GM products do not necessarily need to "escape." It is very VERY easy to make organisms unable to breed with wild strains. The easiest is to only distribute sterile seeds when they are not being planted in a controlled environment (ie an open field rather than a greenhouse). However the downside to this is that farmers are then FORCED to buy new seeds every year. Now while farmers ALREADY tend to buy most or all of their seeds every year, this forced addiction may be unacceptable.
Probably the best way is to make the GM food unable to mate with wild strains. For insance, plant sperm needs to recognize and attach to plant ova. With out this attachment no fertilization can occcur. Fo the mdification is simple. Replace the two genes necessary for this matching with simlar genes from say a grasshopper....Now the GM plants absolutly cannnot coss with the wild strains and mating with grasshoppers won't be viable (:-))
But your BIGGEST oversight is the assumption that nature does it best.
It has been proven that nature is random. Genes are randomly distributed in each generation. Ever now and then a new gene or combination of genes are created that fundamentally changes the way the organism interacts with its environment...it forms a new species.
Just like introducing bull frogs to australia, this new species will wreak havok on the environment until a new balance is found. This is NO different that what would happen if GM plants got out.
Finally, there is a great deal of evidence that genes ARE transfered between completly unrelated species. This is done through bacteria or cross species virus's. A bacteria or virus picks up DNA from it's first host and the 2nd host incorproates it. Now fish DNA IS in soybean DNA. There is a great deal of evidence that suggests that a similar process triggered our evolution from other primates.
While GM food could be dangerous, it is no more dangerous than any other product we create. While removing a gene can cause changes to an organism, they will have to be very minor changes for a the reserachers not to see them. So long as proper percautions are taken...testing the food produced, checking the plants with varius others in a simulated wild situation etc...GM food can be very safe and a boon to society in general.
This post is modded "Score:2, Insightful" !?!?!? If it doesnt change very fast, I don't like Slashdot anymore :(
Do you blame the Global Warming on Ford? Nobody does. Did he play a key role in it? Well, noone can deny that (thought some might argue about it for long if they bothered).
What is dangerous is greed and foolishness. Like Monsanto rushing out to use a largely uncontrolled new technology on a cornerstone of life on Earth to make money for their share holders. And the people who keep whining about the 'ignorant Europeans' and 'eco-freaks' who are just 'paranoid' and claims that such a brilliant invention should just right away replace what put Homo sapiens here in the first place. (It gets disgusting when some claim that they are merely making the money to save the starving African children though.)
We can also argue for a long time if Microsoft and/or Monsanto are 'evil' in the real meaning of the word. But most will agree that developing Open Source software and now GPL GM crops is a very interesting idea at least.
But thank you for reminding me there are people like you out there - if I knew the word for what you are and used it, I'd propably get modded down to -10 for rudeness.
GM crops have tremendous potential in regions such as Africa
Let's not fall for the myth that there is a world food shortage. Crisis, yes; shortage, no. There's actually plenty of food to feed everyone and more, the problem is in distribution and the international markets (note for example, that even at the height of the famine in 1984, Ethiopia was still a net agricultural exporter; 'course you can't feed your population on coffee beans). You're absolutely right that Western governments have rigged the market in favour of their own agribusinesses, but why would you think that the introduction of GM would do anything but make that situation worse? GM is all about locking in control of the food chain, from the field upwards.
It's bad enough that third world countries have to concentrate on growing cash crops for foreigners rather than food for their own people, in order to service their foreign debts. Now you expect them to be additionally burdened with the licence restrictions ("taxes" might be another word) associated with GM. How exactly does it benefit an African farmer if she now has to pay an annual licence fee in order to be allowed to plant the seed she saved from the crop she grew last year?
(Not that I disagree that the EU's agricultural policy is anything other than scandalous. The Common Agricultural Policy is an anachronism from the post-war period that should have been fixed thirty years ago. I would hardly call it "sentimental" ).
My next sig will be ready soon, but subscribers can beat the rush
is nowhere nearly as efficient as helping none of us.
Laws are for people with no friends.
The farmer I mentioned went bankrupt because he had no way of identifying, or removing, the GM strain of crop from his field.
Note that there is a very simple way for Monsanto to identify whether (in this case) there are Roundup-Ready (in this case) crops present - they simply fly over and spray the field with roundup. If the crops die, then the farmer is innocent. If the crops don't die, then the farmer gets sued. Much like the medieval method for identifying witches.
My next sig will be ready soon, but subscribers can beat the rush
"No, you're confusing two things."
Nope, you have it confused.
"The consumer is torn between the desire for ultra-cheap food right now, this instant, and wanting his childrens and maybe unborn grandchildren to be born with the right number of fingers, toes, eyes and ears."
This shows it. While there may be consumers like this out there, they're idiots. It's based upon the nonsense that the food you eat will do those things to you.
I agree completely, those responsible for creating these "terminator seeds" are propogating the most horrible kind of evil. The whole premise is one of ensuring that farmers are unable to harvest their own seeds, and must rely on the "producer" to continue to farm their crops. If these things get out into the wild, it will mean humanity's end, as nations will slowly lose the capability to feed themselves without a multi-national corporation's blessing. Those responsible should die a slow and horrible death. Spreading this type of technology is an equal crime to genocide in my view.