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Ankhetmaatre
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PostPosted: Thu Jun 27, 2013 5:12 pm    Post subject: Reply with quote

Yeah, the 2011 article by Marchant is interesting, if a bit dated at this point. The last sentence seems to reveal the pith of the divide between the "skeptics" and the "believers"...

Quote:
Willerslev agrees, offering a rare olive branch. "I think we will find that the believers have been too uncritical," he says. "But the sceptics have probably been too conservative."


I was very encouraged by the article Sothis linked to above, also authored by Jo Marchant, about ancient DNA from hot climates being sequenced using next gen techniques. She seems to come across as a reasonably fair handed science writer, imo.
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PostPosted: Thu Jun 27, 2013 7:54 pm    Post subject: Reply with quote

SidneyF wrote:
http://www.nature.com/news/2011/110427/full/472404a.html


And this quote from the above goes to the heart of the difficulty:

"But for now, Zink says, "It's like a religious thing. If our papers are reviewed by one of the other groups, you get revisions like 'I don't believe it's possible'. It's hard to argue with that." "

That's why I say it's a lot easier to be a skeptic than a "doer". All it takes is nay-saying.[/quote]

Yet, you fail to include WHY they nay-say, and it's in the very article you quote:

That's no surprise, say sceptics. DNA breaks up over time, at a rate that increases with temperature. After thousands of years in Egypt's hot climate, they say, mummies are extremely unlikely to contain DNA fragments large enough to be amplified by PCR. "Preservation in most Egyptian mummies is clearly bad," says Pääbo, now at the Max Planck Institute for Evolutionary Anthroplogy in Leipzig and a leader in the field. Ancient-DNA researcher Franco Rollo of the University of Camerino in Italy went so far as to test how long mummy DNA might survive. He checked a series of papyrus fragments of various ages, preserved in the similar conditions to the mummies. He estimated that DNA fragments large enough to be identified by PCR — around 90 base pairs long — would have vanished after only around 600 years.

<...>

The sceptics are unmoved. Without highly stringent controls in place, it's impossible to show that any microbial sequences are from ancient DNA and not from related modern microbes, says Gilbert. "How do you know you've got TB and not some other bacterium with a similar DNA sequence?" He and other critics believe that this entire body of research is based on wishful thinking.


In particular, concerning the Zink and Pusch DNA studies of Tutankhamun and his lineage:

His major concern, shared by others, was the method of DNA analysis used. Rather than extracting and sequencing DNA, the team used a technique called genetic fingerprinting, which involves measuring the size of the DNA products that have been amplified by PCR. It is rarely used in ancient-DNA studies, say critics, because without sequence data it is especially difficult to rule out contamination. And on a well-handled mummy such as Tutankhamun, say sceptics, contamination could be rife.

The Tutankhamun team carried out many controls, including replication of the tests by different teams in the two labs and comparing the mummy DNA fingerprints with those of the research team to cross-check for contamination. Zink and Pusch add that the samples were taken from within the mummies' bones where, they say, contaminating DNA should not have reached.

Zink and Pusch think that the mummification process protected the DNA from degrading in the hot tomb by removing water, which is required for the main mechanism of DNA decay, called depurination. Egyptian embalmers dried bodies with natron, a naturally occurring mixture of salts, immediately after death. "The Egyptians really knew how to preserve a body," says Zink. "They got rid of the water very fast." Tutankhamun was also smothered with embalming and anointing materials, thought to contain ingredients such as bitumen, plant oils and beeswax, and Pusch believes it gave the DNA additional protection from the damaging effects of water. Hawass was not directly involved in the DNA research, but he stands by the team's conclusions, saying that the DNA in Egyptian mummies seems to be well preserved.

<...>

Sceptics, however, doubt that there was sufficient DNA left in Tutankhamun for the result to be real. They say that a mummified body would soon soak up any moisture available in the atmosphere, especially into its porous bones. When British archaeologist Howard Carter first opened Tutankhamun's coffins in 1925, he reported that they had been damaged by humidity. But it is difficult for anyone else to replicate the DNA work without permission to access the samples.


...and that, as far as I can see it, is where we are today. In the case of the Pusch and Zink DNA studies of ancient Egyptian mummies, you haven't had the independent verification of their results, which is the standard in the field, since the Egyptian government and antiquities division have not allowed other testing of the DNA.

Until then, the geneticists and other scientists have reason to be sceptical and I don't blame them, considering the possible problems they have outlined.

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PostPosted: Fri Jun 28, 2013 12:28 pm    Post subject: Reply with quote

The thing with the so-called nay-sayers is that they do not just say nay, they tried very hard to do it themselves, but with little to no success. And as scientists such as Paabo, Gilbert, Willerslev are surely just as capable as those who claim great success they have IMO every right to be sceptic. Even more so for as long as the "doers" do not enter into a real discussion.

As for the many of us who are laypersons, we can only hear the arguments brought forward on both sides and try to work out what sounds more plausible.

And I think personally that the methods used in the study of Tut`s family are not reliable enough to guarantee authenticity.
Among the strict criteria to rule out contamination are the demand to use only workspaces where DNA has not been amplified before and to sequence amplified DNA. Neither of these rules has been adhered to. We all know that no sequencing was carried out whatsoever, and the DNA of all eleven mummies was amplified in the one and only lab under the Egyptian museum.
Remember the second lab was built later to double-check the results.
Given that they claim to have tested each locus up to thirty times and took samples from five different locations of each mummy, this makes 150 test runs per mummy and 1650 for all mummies together. Take this number times eight and you have the approximate number of test runs, but only for the autosomal DNA.
Does anyone think they had so many seperate workspaces available in the vaults of the Egyptian museum? Probably not. Surely they will have cleaned up thoroughly between the goes but that`s not quite enough. The rules state unused workspaces.
And because PCR is so sensitive, it might have picked up not only modern contamination, but perhaps also some left-over molecules from a fellow mummy. Because they only used fingerprinting there would be no way to tell
them apart.

But to me the final death blow for any good faith is the "majority rule" described on pages 206/207. As they did not get the same results for every test run, they had to work out which result appeared more often than others which was then recorded as final result. I have tried to find out if this is common practice when dealing with ancient DNA, but apparently it is only used in cloning, not in fingerprinting or sequencing.
This really gives the impression of throwing dice and working out probability.
It does not inspire confidence in the results thus obtained, and this is also hte main reason why they do not share raw data with fellow scientists, as Zink frankly admits.
But everyone has to judge for himself.
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PostPosted: Fri Jun 28, 2013 1:54 pm    Post subject: Reply with quote

I have found that simply assigning scientific jealousy to nay-sayers, especially those with the accomplishments and credibility of Paabo, Willerslev, ect, is extremely short sighted. As I tried to say upthread, scientific methodology is followed for a good reason. Unfortunately, Zink, Pusch, et al, were not able to follow proper extraction or sequencing methodology, through no fault of their own. And, though this is speculation, I don't doubt they were under pressure from Hawass for results.

As an aside, I recently watched one of those awful History Channel docs where someone named Olly Steed and the former curator of the Ciro Museum were inspecting some the royal mummies in a stairwell. God knows why they were in a stairwell but the mummies were open to the air and being handled without gloves or masks. :/
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PostPosted: Fri Jun 28, 2013 2:26 pm    Post subject: Reply with quote

Ankhetmaatre wrote:
I have found that simply assigning scientific jealousy to nay-sayers, especially those with the accomplishments and credibility of Paabo, Willerslev, ect, is extremely short sighted. As I tried to say upthread, scientific methodology is followed for a good reason. Unfortunately, Zink, Pusch, et al, were not able to follow proper extraction or sequencing methodology, through no fault of their own. And, though this is speculation, I don't doubt they were under pressure from Hawass for results.


For which they were willing to risk their professional reputations? And for this, as well? The groundbreaking work has, after all, continued.

http://www.bmj.com/content/345/bmj.e8268

Quote:
As an aside, I recently watched one of those awful History Channel docs where someone named Olly Steed and the former curator of the Ciro Museum were inspecting some the royal mummies in a stairwell. God knows why they were in a stairwell but the mummies were open to the air and being handled without gloves or masks.


And that contaminated the inside of the bones from which the samples were taken--how? What the nay-sayers here are forgetting to mention is that the family relationships and the very identifications of the mummies as corroborated by the DNA study was hardly shocking. As I already mentioned, the mummies checked and balanced one another--and I can't understand how people can fail to understand this simple concept.

IF none of the mummies tested had ever been positively identified, then one might be more justified in questioning the results published in JAMA. However, this is not the case. And all that the team was trying to establish for the purpose of the paper was familial relationships [pathology aside].
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PostPosted: Fri Jun 28, 2013 2:38 pm    Post subject: Reply with quote

This site is starting to make it difficult to type after a couple of paragraphs and, since someone else mentioned encountering the problem in this thread, I have to conclude it isn't just me. Anyway, to continue:

Yuya and Thuya, the oldest generation in the study, do not have any doubt as to their identity. If they passed on their alleles to the Elder Lady, then where is the surprise there? People have long suspected she could be Queen Tiye and even Renate Germer, who questioned this ID in a paper, had to admit that she could be the queen. The mummy of Amenhotep III was identified by men in remote antiquity. It would have been nice to see the DNA of his father, Thutmose IV--but if the alleles of this man and the Elder Lady were passed on to the next generation--again no surprise. The KV55 individual has long been surmised to have been either Akhenaten or a brother, Smenkhkare. The surprise was that the Younger Lady should have proved to be the mother of Tutankhamun, but I don't see how that is unlikely at all. So where are all the errors made by the team that the people here suspect?
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PostPosted: Fri Jun 28, 2013 8:40 pm    Post subject: Reply with quote

Seems to me the quote from neseret's post explains it all pretty succinctly. Did you read it?

As far as contamination via handling goes, yeah, that sort of thing can cause all kinds of contamination depending on HOW the samples are taken. Especially since the sort of situation that tv show evidenced has happened many times before. The devil is in the details.

Enjoy this helpful link: https://en.wikipedia.org/wiki/Scientific_method

Relevant to this discussion...
Quote:

Scientific inquiry is generally intended to be as objective as possible in order to reduce biased interpretations of results. Another basic expectation is to document, archive and share all data and methodology so they are available for careful scrutiny by other scientists, giving them the opportunity to verify results by attempting to reproduce them. This practice, called full disclosure, also allows statistical measures of the reliability of these data to be established (when data is sampled or compared to chance).


SidneyF said
Quote:

Yuya and Thuya, the oldest generation in the study, do not have any doubt as to their identity. If they passed on their alleles to the Elder Lady, then where is the surprise there? People have long suspected she could be Queen Tiye and even Renate Germer, who questioned this ID in a paper, had to admit that she could be the queen. The mummy of Amenhotep III was identified by men in remote antiquity. It would have been nice to see the DNA of his father, Thutmose IV--but if the alleles of this man and the Elder Lady were passed on to the next generation--again no surprise. The KV55 individual has long been surmised to have been either Akhenaten or a brother, Smenkhkare. The surprise was that the Younger Lady should have proved to be the mother of Tutankhamun, but I don't see how that is unlikely at all. So where are all the errors made by the team that the people here suspect?


^ This is classic inductive reasoning https://en.wikipedia.org/wiki/Inductive_reasoning and is the antithesis of scientific method. The fact that all the participants in the DNA testing knew who these individuals were supposed to be is another strike against the findings, unfortunately.


Believe me, I would like it very much if these studies could be more verifiable, but they aren't, yet. Too many steps in proper methodology have been ignored or skimmed over. I admire the work of Zink and Pusch and I'm hopeful because they're still engaged in shoring up the work they started. I think everyone looks forward to seeing it move forward.
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PostPosted: Fri Jun 28, 2013 9:32 pm    Post subject: Reply with quote

no offence to anyone, but with ancient dna being so hard to work with, and being a fairly new science how many of the writers or anyone here are certified to know all the acceptable procedures as well as reading this complex form of data. As for the conditions of the lab that just seems to me as a backup plan if the results came back unacceptable. Makes it to easy to say procedures were not followed so results are no good
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PostPosted: Fri Jun 28, 2013 10:01 pm    Post subject: Reply with quote

[quote="Ankhetmaatre"]

SidneyF said
Quote:
Yuya and Thuya, the oldest generation in the study, do not have any doubt as to their identity. If they passed on their alleles to the Elder Lady, then where is the surprise there? People have long suspected she could be Queen Tiye and even Renate Germer, who questioned this ID in a paper, had to admit that she could be the queen. The mummy of Amenhotep III was identified by men in remote antiquity. It would have been nice to see the DNA of his father, Thutmose IV--but if the alleles of this man and the Elder Lady were passed on to the next generation--again no surprise. The KV55 individual has long been surmised to have been either Akhenaten or a brother, Smenkhkare. The surprise was that the Younger Lady should have proved to be the mother of Tutankhamun, but I don't see how that is unlikely at all. So where are all the errors made by the team that the people here suspect?


Quote:
^ This is classic inductive reasoning https://en.wikipedia.org/wiki/Inductive_reasoning and is the antithesis of scientific method. The fact that all the participants in the DNA testing knew who these individuals were supposed to be is another strike against the findings, unfortunately.


You've got to be kidding. Well, actually, I don't think you are--so that now proves you know nothing about how autosomal DNA works. If you don't have any idea who the people involved are--why test them at all? That's right. That's why, when some female wants to prove paternity, she doesn't have every male in her town tested to see if he could possibly have been the father of her child. She already knows who the candidate or candidates are. Is this easy enough for you? I hope so. Therefore, by the same token, certain mummies [out of a much larger group at Cairo] were selected for testing to augment the other criteria for the identifications of the mummies--or disprove the IDs. "Who were the parents of Tutankhamun" was doubtless a question that was high on the agenda. So, by your reasoning, for which I doubt a name has even been invented to this very day, nobody in any DNA lab should ever know the identity of any subject tested for any reason. Because nobody can count on their honesty, right? POOOF!! There goes Family Tree DNA and like companies with all of their surname projects where people submit their information along with their little cotton swabs. OMG! If they know my surname, they might want to put me in with people who have the same surname! Except the DNA isn't grouped by surnames but by DNA results. That's why there have been many surprises and that's why I'm done with you.
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PostPosted: Fri Jun 28, 2013 11:09 pm    Post subject: Reply with quote

Wow, you really, really don't get it. Oh course SOMEONE knows who the mummies are supposed to be. But the lab scientists are not supposed to know. Yes.

You know what? Never mind. Enjoy being laughed at by the rest of the scientific world.
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PostPosted: Fri Jun 28, 2013 11:24 pm    Post subject: Reply with quote

herper wrote:
no offence to anyone, but with ancient dna being so hard to work with, and being a fairly new science how many of the writers or anyone here are certified to know all the acceptable procedures as well as reading this complex form of data.


I would say among the naysayers--zero. Here is the background of the most vociferous, Ankhetmaatre. And I quote:

Quote:
Though I am an artist by trade a particular interest of mine is ancient art (obviously), flora and clothing. I've attempted to recreate a number of Ancient Egyptian garments over the years with good success and have a number of ideas about how many of their artists' stylizations were actually realized in cloth. I'm very interested in any and all links to images of original garments, btw. The stories of individual artists and craftsmen, what little we have of them, are endlessly fascinating to me as well, from Imhotep and Akhenaton to the artisans of Dier el Madina.


That's why they say people are best off sticking to their own area of study.
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PostPosted: Sat Jun 29, 2013 2:33 am    Post subject: Reply with quote

It isn't too difficult to make you defensive, is it, Sidney? Too bad for that.

I am an artist, who happens to be a conservationist. That means I conserve art for museums. I have even sent a number of things to various labs to be analyzed so I know something about the methods used in those processes. It's one of the reasons I talk to people at Harvard on a regular basis.

I'm sorry your insecurities make it so difficult for you to enter onto a discussion without becoming angry. The fact that you seem to want to be insulting and petty is annoying but, ultimately, not my problem. Good luck in the real world with that attitude.
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PostPosted: Sat Jun 29, 2013 2:36 am    Post subject: Reply with quote

Oh, and I'm actually not a nay-sayer. I'm just honest. Wink
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PostPosted: Sat Jun 29, 2013 6:16 pm    Post subject: Reply with quote

SidneyF wrote:
Ankhetmaatre wrote:
I have found that simply assigning scientific jealousy to nay-sayers, especially those with the accomplishments and credibility of Paabo, Willerslev, ect, is extremely short sighted. As I tried to say upthread, scientific methodology is followed for a good reason. Unfortunately, Zink, Pusch, et al, were not able to follow proper extraction or sequencing methodology, through no fault of their own. And, though this is speculation, I don't doubt they were under pressure from Hawass for results.


For which they were willing to risk their professional reputations? And for this, as well? The groundbreaking work has, after all, continued.

http://www.bmj.com/content/345/bmj.e8268


Have you read this article?

First, the article is co-authored with Hawass and mainly members of the Egyptian Museum DNA research group, with 12 of he 14 authors being the same authors of the 2010 JAMA study.

Secondly, and this is in the article itself, "Z[ahi]H[awass], ... YZG[ad], C[arsten] P[usch],, and ARZ[ink) designed the study, analysed the data, and drafted the manuscript." These are also the same authors who designed, analysed and drafted the 2010 JAMA study.

The only new author in the 2012 study is Somaia Ismail, professor of molecular biology, who happens to work for BOTH Cairo University and the DNA Research Lab inside the Egyptian Museum.

Additional information shows other interests involved: "Funding: This study was supported by Discovery Channel and Brando Quilici productions and the Landesgraduierten-Förderung Tübingen."

They go one to note that "...Competing interests: All authors have completed the ICMJE uniform disclosure form at www.icmje.org/coi_disclosure.pdf (available on request from the corresponding author) and declare: support from Discovery Channel and Brando Quilici productions and the Landesgraduierten-Förderung Tübingen for the submitted work; no financial relationships with any companies that might have an interest in the submitted work in the previous three years; no other relationships or activities that could appear to have influenced the submitted work."

I see this as rather odd, as these are the same funding sources as the 2010 Hawass, Gad, et al. JAMA genetic study. So, how can they possibly say that there has been "... no financial relationships with any companies that might have an interest in the submitted work in the previous three years; no other relationships or activities that could appear to have influenced the submitted work." Of course this relationship is with companies that do have a vested interest in the outcome: how can they say otherwise since the very same companies also funded the 2010 study?

In the Methods area, they note the very same problems that have been discussed in this thread, to wit:

Bone sampling was done under sterile conditions in a dedicated room of the Egyptian Museum in Cairo. All staff involved in the sampling wore protective clothing, sterile gloves, and facemasks to prevent exogenous contamination. DNA extraction and purification were performed according to our previously published protocols in a dedicated laboratory in the Egyptian Museum, and replicated in a second laboratory at the Faculty of medicine in Cairo University. (citing the 2010 Hawass, Gad, et al study)

So, there has been no change or variation of methods, the funding sources, who have vested interests, remain the same and the authors, who have reason to claim their work is valid, are using the very same methods to derive the DNA as in the 2010 study, notably without independent verification in a non-Egyptian museum/university genetics lab.

So, as I said before, we are in something of a stand-off to claim the Pusch/Zink DNA-typing method is reliable and/or has "continued" in its work when the 2012 study still suffers from the same criticism as the 2010 AMA study.

Until these results can be replicated in a lab separate from the Egyptian Museum/University of Cairo labs, which is standard in the field of genetics - and this usually means by a lab outside of Egypt - then geneticists and other scientists have reason to be sceptical.
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PostPosted: Sat Jun 29, 2013 7:31 pm    Post subject: Reply with quote

[quote="neseret"]

Quote:
Have you read this article?


You know I have.

Quote:
First, the article is co-authored with Hawass and mainly members of the Egyptian Museum DNA research group, with 12 of he 14 authors being the same authors of the 2010 JAMA study.


Quote:
Secondly, and this is in the article itself, "Z[ahi]H[awass], ... YZG[ad], C[arsten] P[usch],, and ARZ[ink) designed the study, analysed the data, and drafted the manuscript." These are also the same authors who designed, analysed and drafted the 2010 JAMA study
.

And your point is? Are you saying they're not capable of doing the work?

Quote:
The only new author in the 2012 study is Somaia Ismail, professor of molecular biology, who happens to work for BOTH Cairo University and the DNA Research Lab inside the Egyptian Museum.


And?


Quote:
Until these results can be replicated in a lab separate from the Egyptian Museum/University of Cairo labs, which is standard in the field of genetics - and this usually means by a lab outside of Egypt - then geneticists and other scientists have reason to be sceptical.


Well, the "field" would certainly be surprised to hear of it. It's not standard at all. While the teams working on ancient remains have often been international, they did not submit their samples to other teams to be confirmed. For example, what separate lab replicated the results for Otzi the Iceman? However, with all the brouhaha nowadays that might not be such a bad idea. On the other hand, what if the second team isn't as capable as the first and can't replicate? Does that necessarily mean the first team was making false claims?
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