Romanov Imperial Bones Revisited: why
does doubt remain about who is buried in the
“The truth about the death of the Tsar – will offer the truth about the suffering of Russia.”
Margarita Nelipa and Helen Azar
The household clock chimed the hour past midnight and soon the Commandant of the house on Voznesenskii Prospekt, methodically awoke the residents and requested that they dress and follow him downstairs. The sickly young boy racked by the pain of his congenital condition, now permanently disabled and unable to walk, had to be assisted by his father, who cautiously carried him down the dimly lit flight of stairs to the lower level of the residence. The rest of the family, accompanied by the boy’s tired mother and four youthful sisters followed, as did the family’s remaining staff. Escorted by the Chekist guards bearing weapons, none had any idea where this unexpected nocturnal journey would lead. Decades would pass before the world would learn about this Commandant and the role he was about to enact, ensuring that history will never forget the name of Yakov Yurovsky.
Within several minutes the deed was done, and the walls
spattered with fresh blood and stray bullets became silent witnesses, which
would give their testimony within days as to what had transpired whilst the
The world was forced to wait seventy years, when under the umbrella of Mikhail Gorbachev’s Glasnost’ (a policy of openness), the Soviet media announced via an interview with Geli Ryabov, in Moskovskie Novosti (Moscow News)  on 12 April, 1989, that the Imperial remains had been found. The official Soviet blackout for any public discussion of the real destiny of the last Russian Imperial Family was slowly melting away. Two years later, on 11 July 1991, after the collapse of the Soviet regime, Boris Yeltsin’s government, at the request of the Ekaterinburg regional Governor, Edvard Rossel, sanctioned that the discovery of these remains could come to light.
Clearly, years had to pass to ensure that Dr Alexander Avdonin and Geli Ryabov’s mutual secret of the grave’s location, which they originally found in 1978, could be safely revealed. It was time that the bone segments and partial skulls submerged in the murky depths of the mineshaft could receive the customary Orthodox funeral service and be reburied.
In 1998, Dr Avdonin facilitated the safe return of the skeletal remains back to St. Petersburg, to join in death the other Imperial remains traditionally interred in the St. Peter and St. Paul Cathedral.
It was exactly to the day eighty years after their assassination. The official mourners included members of the Romanov House and world dignitaries who witnessed the Orthodox blessing and burial customary for their Imperial status.
No one could foresee at the time that the official forensic uplift in 1991 would signal not a journey towards closure, but instead a long tedious road that would begin to tumble into controversy.
Much of this controversy was led by a group of Russian
descendants living outside of
The following discussion shall address why we, as professional scientists, contend that the remains buried in St Peter and Paul Cathedral are indeed those of the last Russian Imperial Family, and that the objections brought forth to argue to the contrary are not scientifically valid.
Why is there still concern about the authenticity of the Imperial remains?
Almost ninety years have passed, and despite the series of
confirmationary scientific and anthropologic assessments carefully conducted in
various accredited laboratories around the world, doubt continues to be cast by
a group of private individuals who reside outside of
RECA’s reasoning is not based on scientific merit but on
political considerations. In one statement, one of its members, Peter
Koltypin-Wallovsky claimed that “the
bones were part of a fabricated case concocted by the communist regime and
perpetuated by Yeltsin’s government. … they created these remains”, he stated. The group strongly identifies with the
In 1993, this émigré group, who it must be stressed, do
not represent the Russian communities living in exile around the world,
made their presence known in Russia, by sending a letter to the Deputy Premier
of Russia, Yuri Yarov, stating that “We
suppose that the other bones were put there in 1979 so that it was possible to
fake the recovery of the remains in July 1991.”
The logic of this presumption escapes comprehension. The head of the Russian
Prosecutor’s Office, Vladimir Soloviev, formerly head of the criminal juridical
RECA is convinced that the DNA analyses conducted by Dr
Peter Gill of the U. K. Forensic Science Service (FSS) and his team of DNA
experts, erred with their authentication procedure. RECA reached this decision, based on the
assertions that the “Yurovsky note of 1922” was created by another individual,
It was believed that because Yurovsky was poorly educated he was incapable of
writing the document attributed to him. We however contend that it is more than
plausible that Yurovsky dictated what his participation was to
Pokrovsky, to ensure that a permanent archival record would be preserved. Such
a consideration is not an unrealistic scenario, because Pokrovsky who was appointed by Lenin to take charge of
the Central Archives in
First Stage: Authentication of the Romanov Bones – The Macroscopic Examination
Initially, the skulls were examined by Dr Sergei Abramov - a Moscow-based forensic anthropologist. Using complex computer superimposition imagery he compared each skull with enlarged imperial facial photographs, which allowed him to confirm in 1992 that the skulls belonged to Emperor Nicholas II, his wife, the Tsaritsa Alexandra Fedorovna and three of their daughters, Grand Duchesses – Anastasia, Tatyana and Olga. Abramov with his colleagues published their results in English in Anatomical Record in 2001.
During the morphologic phase of their study, they compared 60 unrelated skulls (controls) to verify that the mathematical models employed would differentiate the different cranioscopic characteristics that define the skull contours and lines. Four of the skulls showed “remarkable stable similarity.” By the use of photographic superimposition as a second identification process; Nicholas II and Alexandra Fedorovna were identified by both methods.
To ensure global acceptance of these remains, a team lead by the American forensic anthropologist, Dr William Maples arrived in Ekaterinburg by invitation of the Russians, and examined each of the nine skeletons laid out in the town morgue. Dr Maples and his team quickly confirmed that the skeletal remains bearing gunshot wounds, tagged by number ID only, were authentic.
The physical human determinants: gender, age, weight, height and facial bone structures matched exactly. In addition, the teeth embedded in the skulls conformed to the type of dental treatment that was afforded to persons of aristocratic status. Some of the teeth contained gold and platinum dental work, – a luxury that would have been impossible for random murdered victims in a distant Siberian town. Another telling aspect that helped to confirm that the scientists had the correct set of remains was that one of the skulls had no teeth. Sokolov’s Report revealed that Dr Evgenii Botkin’s dentures were found in the Four Brothers field by the first investigator back in 1919, which was confirmed at the time of the find by Pierre Gillard, one of the tutors to the Imperial children. The absence of dentition on one of the adult skulls immediately signaled to Dr Maples that it belonged to Lieb-Medik Botkin. The skeletal remains of one of the younger grand duchesses and the Tsesarevich Alexei were not among the remains exhumed in 1991.
Both the Russians and
Clearly, the nine skeletons correlate exactly with the profile of the individual Imperial family members and their staff found in a single grave proximate to the place of execution, and logic tells us that it would be impossible to duplicate the same physical criteria artificially. RECA’s later public statement that the remains buried in the Fortress belong to “unknown victims of the Russian Civil War,” was clearly misguided.
But the most important aspect of the identification process at this stage was to conduct DNA profiles and compare them with living relatives of the assassinated Imperial family.
Second Stage: Authentication of the Romanov Remains – The First DNA Analysis
Briefly mtDNA analyses examine DNA extracted from cellular
organelles called mitochondria. Since older biological forensic samples may
often lack nucleated cellular material, bones, hair and teeth can be analyzed
using mtDNA profiles.
DNA analysis is a powerful diagnostic tool that is used to identify individuals
as well as familial relationships. Equally it is perfect for forensic
investigations and is the standard procedure to identify victims of disasters
The human mitochondrial genome contains 16,569 nucleotide pairs, which was been completely mapped and sequenced in 2003. The sequences of nucleotide bases are classed as amino acids: adenine, cytosine, guanine and thymine, and are commonly abbreviated: A, C, G and T. A few mutations that include single base changes (or point mutations) have been described in scientific literature. Obviously, if the mtDNA mutation is rare it will enhance the authentication inquiry. It was fortuitous that the Romanov case did demonstrate a rare mutation and that the same mutation was observed in three living relatives.
The Molecular Investigation Begins
The Russian forensic medical geneticist and Director of the
Russian DNA Finger Printing Center,
Professor Pavel Ivanov, was given
consent by Russia’s Chief Medical Examiner, Dr Vladislav Plaskin to take the
Romanov tissue samples to England where the technology to perform sophisticated
mtDNA tests was available. This was in September, 1992. Jointly with Dr
Peter Gill, and Dr Erica Hagelberg, a Norwegian specializing in
molecular genetics of ancient bones; they spent over 10 months analyzing the
bone tissue at molecular level.
During the course of their extensive studies on 29 September 1993 it was
announced in Russkaya Meditsinskaya
Gazeta (Russian Medical Newspaper) that the bone fragments which Professor
Pavel Ivanov took to
In 1994 the international team were in the position
to formally publish their laboratory results in Nature Genetics,
one of the most prestigious independently peer reviewed journals in the
scientific world. These results conclusively confirmed that the DNA profiles
extracted from the femurs matched exactly the DNA profiles assessed from blood
samples donated from living descendants that included the Duke of Edinburgh
from Alexandra Fedorovna’s side; and Countess Ksenia Sheremetyeva-Sfiris
(Nicholas’ sister Ksenia’s great-granddaughter), from Nicholas II’s side of the
Curiously at the time, the Emperor’s nephew, Tihon Kulikovskii (the son of
Nikolai’s other sister Olga), categorically refused to donate his blood or hair
samples to either Dr Gill or Professor Ivanov on “political grounds”,
which related to his belief that a “Russian
man …working in England, which was so cruel to the Tsar”, was not compatible
with his own philosophy. Later his widow added that only a Russian
Orthodox Commission would be acceptable to her.
Shortly before Tihon’s death from a cardiac condition he did offer a blood
sample to the
The Romanov study was conducted in two stages: first from 15 September 1992 to 1993 with Gill in
Englandat the Forensic Science Services Laboratory (FSS), while the second stage was conducted in by Dr Thomas Parsons at the U. S. Armed Forces DNA Identification Laboratory (AFDIL) jointly with Professor Ivanov, from 4 June to 9 September 1995. [Table 1] Between August 1993 and January 1998 all analyses were under the control of the Soloviev Commission (see below). United States
The Second Stage of the DNA Analyses
Additional tests were performed at this second stage to compare mtDNA derived from the putative remains of Emperor Nicholas II, to that of his brother, Grand Duke George Alexandrovich, who died in 1899 at the age of 28. It took almost two years to receive permission from the St. Petersburg Orthodox Church to exhume George’s remains in order to gain “further insight into the occurrence of the mutational variance of the Emperor’s maternal lineage”. His remains were finally exhumed, in July 1994, under the authority of the late St. Petersburg Mayor, Anatoli Sobchak, against Mrs. Kulikovskii’s wishes. Fundamentally the latter was annoyed that no Romanov family member was asked to attend the exhumation. She expressed distrust of Dr Ivanov “Did he take the bones from Ekaterinburg to compare to the Grand Duke’s, or did he take another bone of George’s and claim it was the Ekaterinburg remains?
With this second strand of investigations, Ivanov was able to advance his DNA assessment, and in 1996 he co-authored a second article in Nature Genetics. This publication discussed the comparative DNA profile analyses between the exhumed remains of Grand Duke George and Nicholas II. In this laboratory study, the mtDNA from the same left femur and tibia bone samples that were used previously in England were re-assayed, in America, independently, using this second laboratory’s own set of protocols and resources. The advance here was that the American team was now able to correlate the mtDNA profiles between George’s left femur and tibia to the second set of bones putatively belonging to his brother, Nicholas II. This careful process also excluded all possibility of using bones from the same individual. [See Table 1]
Table 1: Rare heteroplasmy mutations in the Romanov Family and their relatives
Donor tissue samples
independently by FSS in
Heteroplasmy or fixation at 16169 position on mtDNA sequence
Grand Duke George
Ksenya Sheremetyeva-Sfiris [repeat]
Nicholas II [repeat]
Ivanov identified and confirmed that both the males had “absolute positional identity of mtDNA” that demonstrated a rare genetic mutation referred to as heteroplasmy, this time by an independent laboratory that confirmed the same exact sequences which the Gill laboratory obtained in 1994.
According to Gill, an expert in heteroplasmy, this condition is “a transient event” in genetic terms. Gill expected that a fixation (i.e. with the loss of one or the other of the fluctuating bases it will become either a permanent “C”, or a permanent “T” at that genomic position) would occur in a couple of generations. If the mutation did not skip a generation then all of human population would demonstrate heteroplasmy, which in reality is not the case.
Professor Ivanov’s finding was important for a second reason, in that the heteroplasmy was not simply a random event; but was shown to run through the maternal line on Nicholas’ side. In other words, the brothers demonstrated the same heteroplasmic mutation in exactly the same position of their individual DNA sequences. From an analytical point of view - this type of identification application was demonstrated for the very first time to the world. [See Table 2]
Table 2: Romanov Familial Relationships
Combining the DNA profiles and the rare heteroplasmy
chromatographic sequencing with all the previous scientific endeavors conducted
It is noteworthy to mention that both strands of the DNA investigations of the Imperial remains were assayed and independently identified by two of the most internationally acclaimed DNA facilities in the world. Their collaboration added strength to the veracity of the final result.
Ivanov also brought a fragment of the bloodstained handkerchief, (not a bandage as Mrs. Kulikovskaya described) that originated from Nicholas’ visit to Otsu (see below), but because they found traces of foreign DNA they concluded that this fragment was compromised when too many unknown persons came into contact with the handkerchief during the period of its handling and later preservation as a museum exhibit. On this basis the sample was excluded from further research. RECA was quick to proclaim that the size of the sample must have been inadequate in that: “If he was a professional scientist used to doing DNA testing, why would they have taken too small a sample?” This handkerchief would cause enormous controversy in a few years, when it came into the hands of RECA (see below).
One would have expected that this would have been the end of
the investigative journey, but as it turned out there was more to come to
impede the Imperial Family from being buried in
The Russian Government Commission Assumes Control
from the Public Prosecutor’s Office assumed control of the Romanov
investigation in August 1993 as if it was a criminal investigation. His
task was to co-ordinate activities from
The Commission also authorized the exhumation of Grand Duke George in St. Petersburg in the previous year, in July 1994,  and also gave permission for an outside organization (RECA), which were not affiliated directly with the Russian Commission, to facilitate an independent analysis to be conducted by Professor Rogaev in Canada (see below) in 1995.
Essentially the Commission’s brief was to collate and weigh
all the investigative evidence before them, and then present its Report to the
President of Russia.
The Commission comprised all interested parties, including Dr Avdonin, the
playwright Edvard Radzinsky, Prince A. Golitsyn (President of the Moscow
Sergei Mironenko (the chief Archivist of the
The goal of the Commission was to authenticate the Ekaterinburg remains for the specific purpose that the Imperial Family and their staff could be re-buried.
At this point, controversy arose as to who should test the
Grand Duke George’s remains. Soloviev desired that Dr Gill carry out the
analysis, but in the end Ivanov had to go to
RECA aggressively challenged the investigative efforts of
Soloviev’s Commission. All their arguments were centered not on the results
obtained, but on their steadfast political ideology, or as Massie eluded, –
based on personality clashes and their hatred of the previous Soviet
Kulikovskaya complained that no Romanovs were represented
and thus RECA established themselves as the global monitor in connection to all
decisions that emanated from
Thus, to appease RECA, Soloviev asked Mrs. Kulikovskaya
which DNA specialist she wished to nominate so that she could be convinced that
the remains were authentic. She chose Dr Evgenii Rogaev,
who was at the time undertaking research into Alzheimer’s disease as a visiting
professor at the
The Third DNA Analysis: on behalf of RECA for the Soloviev Commission
Thus the Commission appointed Professor Evgenii Rogaev, a molecular geneticist at the Psychiatric Health Center (RAMN) to conduct an independent analysis on Tihon Kulikovskii’s blood sample.
Evgenii Rogaev and his team analyzed polymorphic regions of mtDNA obtained from the blood sample of Tihon Kulikovskii. He also looked at the mtDNA sequences of other matrilineal descendants of Louise Hesse-Cassel (great-great-grandson and great-great-granddaughter).
Previously, Dr Peter Gill observed a C/T polymorphism at position 16169 in the mtDNA sequence of Nicholas that proved to be an identical condition later seen in the mtDNA of his brother George. This sequence was compared to the mtDNA from two modern day descendants of the same maternal line: the Duke of Fife and Countess Ksenia. The mtDNA of both of these individuals exhibited fixation (“C”) at position 16169. Tihon Kulikovskii - the son of the Nicholas’ sister, had an identical mtDNA sequence except that the fixation at 16169 was with the base “T”. This would be expected with the C/T heteroplasmy present in Nicholas and George. The obvious conclusion is that independent mutations at 16169, initially exhibited as heteroplasmy, occurred in the lineage of Louise Hesse-Cassel.
The mtDNA sequence from Kulikovskii showed an “almost complete match” with the mtDNA sequences obtained by Gill from the putative remains of Nicholas II. The difference was a single nucleotide at the position 16169. At this position, a “C” was found in mtDNA of Kulikovskii, where a C/T was in the mtDNA from the putative remains of Nicholas II. In the mtDNA sequence from two other descendants of Louise Hesse-Cassel (the great-great-grandson and the great-great-granddaughter) a “T” was found in the same position (16169). These data confirmed that independent mutations took place in the maternal lineage of Louise Hesse-Cassel's descendants, and/or a mutation leading to heteroplasmy in the said lineage.
Since all the Ekaterinburg remains were secured in
The Fourth DNA Analysis: on behalf of the Russian Government Commission
On 29 January 1998, Rogaev
submitted a second report to the Commission,
except this time he was given the opportunity to re-assess Kulikovskii’s blood
sample back in
Rogaev’s analysis once again
confirmed the results of Dr Gill and Professor Ivanov, in that the remains were
those of the Romanov Imperial Family.
Thus, in a short space of time, Rogaev’s definitive findings became part of the Commission’s final report to Yeltsin; just months before the re-burial was take place. The Soloviev Commission wanted to ensure that every possible avenue was examined and verified before submitting their final unanimous recommendation to the government.
The Commission’s work is complete
The Commission confirmed that five of the skeletal remains found in Ekaterinburg in a common grave belonged to the same family – a father, mother and three daughters. Those remains were established without reservation to be those of the Imperial Family, who’s DNA was analyzed by numerous accredited laboratories around the world. Their only goal was the provision of all results of their five-year analyses without favor or prejudice.
According to “Protocol # 9”, dated 30 January, 1998, each member of that Commission (22 persons) endorsed the re-burial, of which 16 members gave their preference for the re-burial to be the traditional St. Petersburg Fortress Cathedral, rather than Ekaterinburg
(4 votes) or the Novospassky Monastery (1 vote).
Boris Nemtsov presiding over the Government Commission for the Identification and Re-burial of the remains of the Imperial Family affirmed the Commission’s findings on 30 January, 1998, and directed its recommendation to the Russian President Boris Yeltsin to facilitate their re-burial in the Fortress Cathedral. On 27 February, 1998, the Russian Government unanimously ratified the Commission’s decision to conduct a re-burial.
Finally, over the objections of the Moscow Orthodox Church,
the decision was announced that the Imperial family and their loyal staff could
be re-buried in
The present head of the Romanov family (en exile), Prince Nicholas, insisted that all remains, not just the Romanovs had to be buried together in the Fortress Cathedral, which although against past imperial tradition, the circumstances were extraordinary. It was a gesture to demonstrate a measure of respect towards the faithful staff who not only voluntarily chose to follow the Imperial family into exile, but also died with them. This wish was granted. It was to be the end of a long journey home, but the political rumblings were only beginning.
Tombs of the last Russian
Imperial Family and their retainers inside the St Peter and Paul Cathedral,
Plaques for Nicholas II and Alexandra Fedorovna
(Photo by Margarita Nelipa)
Certainty Begins to Crumble
Any reasonable observer might believe that Rogaev’s analysis would have allayed RECA’s concerns, but this was no so. Before the Russian Government Commission, Mrs. Kulikovskaya announced that Rogaev’s results did not correlate with the Ekaterinburg remains! This declaration was made despite Rogaev’s publication in the Russian genetics journal Genetika, in 1996, which agreed with the original results obtained by Dr Gill and Professor Ivanov. Mrs. Kulikovskaya expanded why she considered that there was incompatibility of results in her interview to a Russian Orthodox magazine. She stated that “Ivanov’s word “mutation … was simply his personal theory” and that “Tihon’s blood was “one figure off, and lacking one figure means that it is not a match.” It must be emphasized that Mrs. Kulikovskaya is not a scientist. Rogaev actually said that “one nucleotide”, which is a single amino acid (an organic compound), that “differed from the profile of Nicholas II.” [Table 3]
Skeptical RECA challenged the Commission head on by
responding that there were scientists who questioned the manner of the DNA
tests. They questioned why either the Dowager Empress, who was buried in
Part of the answer lies in two conflicting assumptions:
1. When Nicholas was Tsesarevich,
during his visit to
According to Alexander Bokhanov’s recent biography of
Nicholas II – the event at
Now let us examine the Medical Report of the incident that was received by Alexander III on 29 April (Old Style) 1891:
“The lesions sustained are as follows: The first, or occipital-parietal, wound is linear in form, measuring nine centimeters, with torn edges, and has penetrated the whole thickness of the skin down to the bone; it is situated in the area of the right parietal bone six centimeters from the upper edge of the ear, extending slightly downwards. Furthermore, vessels of the nape and temporal arteries have been cut. At the rear of the wound, the parietal bone has lost about a centimeter of periosteum, consistent with a blow from a sharp saber. The second, or front parietal, wound is situated some six centimeters higher than the first and runs almost parallel, being ten centimeters in length; it has penetrated right through the skin down to the bone, and occupies the area of the parietal and part of the frontal bone.
… While cleaning the second would, I removed a wedge-shaped fragment, about two and a half centimeters long, which was in the clots of blood. The fragment was of the thickness of ordinary writing paper.”
The incident did not necessitate hospitalization as would have been expected had the sword penetrated the cranium – an injury that would have exposed Nicholas’ brain. Such a scenario would have proved fatal in the absence of ongoing medical attention in the days when antibiotics were unavailable. In fact, the Tsesarevich unperturbed went on to catch the train soon after. Clearly the injury was nothing more than minor and was confirmed by the Medical Report of 1891, which specifically stated that both wounds went “to the bone”.
Soloviev made a very prudent observation, in that the resultant trauma to the skull only affected the periosteum – which was the “fragment” described when the wound was cleaned. The peritoneum is a fibrous protective membrane, which is highly vascularized and forms the external layer covering the cranium. It certainly is not bone! The periosteum, because it is soft tissue, would have not been preserved for eighty years. It would have degraded – initially because of the sulfuric acid that was poured over all the faces and secondly, as a consequence of natural postmortem decomposition of all soft tissue. The fact that it was a paper thin “fragment” excludes it to be anything other than a membrane, because the normal cranium has an average thickness of 4.0 mm. What this all clearly means is that Nicholas’ skull would not have any evidence of past bone repair caused by the Otsu incident.
2. RECA firmly believes that Nicholas was beheaded at some
point during the disposal of Nicholas’ corpse, and that the head was taken to
Yakov Sverdlov in
Another part of the answer was provided by the criminologist Vladimir Soloviev, who believed that RECA was misguided in believing Sokolov’s 1919 forensic investigations that stipulated that all the corpses were “totally burned and destroyed.” Since Sokolov never found any bodies, and in view of the fact that the only human remains he managed to find was a “severed middle-aged female manicured finger”, he was not in the investigative position to contemplate the real circumstances of their death.
It is obvious that RECA is very confused; maintaining their
misguided presumption that any decisions emanating out of
What are the Scientific Contradictions?
Meanwhile, to obtain results that accorded with their own philosophy, RECA privately sought and employed selected laboratory personnel to provide contradictory evidence, alleging it to be more scientifically valid than the results obtained by Gill and his team of experts, and the more recent results provided by Pavel Ivanov.
Following the ratification of the Russian Commission’s report, Olga Kulikovskaya decided that only a judicial court had the legal authority to determine the authenticity of the bones, and not the Russian Procurator.
Outside the Russian Government Commission – Privacy is assured
In 1997 Professor Tatsuo Nagai apparently
The Imperial family was authenticated and buried but, Popov, after meeting Nagai at a conference, in direct contravention of the Procurator’s authority; suggested that Nagai collaborate with him to reassess the Imperial remains. This mutual agreement transpired soon after all the Soloviev investigations were completed.
The paper that Nagai and Popov published jointly in 1999 in a Japanese journal called “Igaku to Seibutsugaku” was never translated into either English or Russian, which would have been prudent, since their results affected Russian and English speaking investigators. This data remains obscure since it was only published in Japanese hence is not readily searchable in scientific databases (which can only sense Latin characters). Therefore it also continues to be unavailable to a wider audience. We had this paper translated into English (with thanks to Mr. Junichi Hayashi) and will summarize it here.
The first thing that was pointed out to us by our Japanese translator was that there was a discrepancy between the Japanese title and the English translation. The Japanese title reads: “DNA identification of Georgij Romanov, a direct [genetic] elder brother of Nicholas II, the last Russian Tsar: Sequence of mitochondrial DNA” [See Figure 1]. The English title, significantly, does not use the term “elder brother”: “DNA identification of Georgij Romanovs, a direct brother of the Russian Tsar Nicolas II: Sequence of mitochondrial DNA.” In the Japanese language, unlike in English and most other languages, the terms for “younger brother” and “older brother” are completely different from each other. The Japanese title in this case specifically refers to the “elder brother”. In fact, the very first sentence states that: “Georgij [sic] Romanov was the real elder brother of the Russian Empire's last Tsar Nicholas II, but he had tuberculosis and was sick and weak, so his real younger brother Nicholas II became the Russian Tsar”. This historic inaccuracy is a minor point indeed but it did set the tone of their investigations with such an unfortunate start.
The first paragraph introduces the reader to the belief that although George was the elder brother, he didn’t become Tsar [Emperor] because “he had tuberculosis and was feeble”. The historic truth was that Nicholas was the elder brother (born in 1867), and George was the younger brother (born in 1871). Nicholas as the eldest son was always heir to the Russian throne. Russian history in fact has proven in the past that no matter how “feeble” a Tsar was he still reigned until his death.
Figure 1: The Japanese title of the Nagai/Popov 1999 paper. The title clearly includes the Japanese words “real elder brother Georgij [sic] Romanov”. Popov’s name appears in English while Nagai’s is in Japanese characters (rendering it unsearchable under the latter’s name).
For the scientific aspects of this study, the Nagai/Popov team analyzed the mtDNA sequence from a sample of hair, allegedly extracted from “Georgij [sic] Romanov”. They compared their results to those published by Ivanov in 1995, and observed “significant differences” at 13 positions: 320, 16093, 16126, 16169, 16223, 16278, 16294, 16296, 16298, 16325, 16327, 16356, and 16362. At all other positions their data matched Ivanov's profiles. They did not find C/T heteroplasmy at the position 16169, but did find it instead at 7 other positions: 16093, 16278, 16298, 16325, 16327, 16356, and 16362. They were unable to propose the cause for this discrepancy.
Overall, Nagai and Popov’s results make very little scientific sense, which may explain why they were never publicized and/or submitted for peer review for possible publication and translation in any international scientific journals that are accessible to the global scientific community. Such a standard expectation is the normal outcome for all legitimate scientific endeavors. It not only facilitates exposure of laboratory research conducted around the world, but promotes the evaluation of those results, especially if they may appear controversial. Essentially peer review is a technique that allows critical scrutiny of any investigator’s sampling, methodology and the conclusions reached, by a panel of experts in the field. Once the article is accepted for publication it then permits others to repeat or question the published results. We contend that such an evaluation would not have been in the best interest of Drs Nagai and Popov, for reasons, which will become more apparent in our discussions below.
What Nagai and Popov’s work actually revealed to us was that the samples they used were either contaminated with foreign DNA, (introduced by inappropriate handling of the tissue prior to any evaluation) - or that the samples belonged to an unknown individual. Furthermore, since there was no “chain of custody” described in their discussion, the question about the authenticity of their sample is open to doubt.
This second comment embedded in their introduction we found to be rather puzzling, and we shall allow you to judge the quality of their presentation:
“… Many still doubt the authenticity of the results. Because of this, Tsar
Nicholas II, and his family, and the doctor’s remains were interred in Peter
and Paul Cathedra (
RECA steps in to contradict the Soloviev Commission and Russian Government
According to Mrs. Kulikovskaya,
RECA appointed Tatsuo Nagai to conduct additional DNA tests after the Soloviev Commission
released their Final Report in1998 to the Russian Government
because of his previous collaboration with Popov in
Kulikovskaya claimed that in February 2001, Nagai expressed
interest in coming to
According to their abstract (#
Nagai and Popov used “the usual
methods” to examine mtDNA sequence from a sample believed to be Nicholas
II’s perspiration stain (from his uniform), and from hair/nail/bone samples
which belonged to Grand Duke George (Nicholas’s brother), as well as a blood
sample from Tihon Kulikovskii (Nicholas’s and George’s nephew). Important note:
scientists do not accept the vague expression “usual methods”. A complete description
of the “Methodology” is normally mandatory. The reason for this information is
very clear – it enables investigators critical information as to how a sample
was processed in that laboratory. If there is a fundamental query, then the
same methodology can be utilized by another laboratory to ensure repeatability
of results. The results Nagai obtained were reported by
him to be “similar to those reported by
Gill and Ivanov, except that heteroplasmy (C/T) was not found at the base
position 16169”. Nagai seems to have attached great importance to the
latter and goes on to state in the same abstract: “thus arises [sic] the question of whose bone was examined by Pater
[sic] Gill et al. and Pavel L. Ivanov et al. Who is buried in the grave of the
Peter and Paul Cathedral at [sic]
One of the more disturbing
aspects of this particular project was that nowhere in the abstract there was
there any mention of the “7 heteroplasmies” or 13 other discrepancies in the mtDNA sequence previously
“discovered” by Nagai and Popov in 1999!
Effectively, Nagai and Popov were not able to match their own previous results using the same hair sample, nor were they able to match their previous results to Tihon Kulikovskii’s mtDNA. This should have alerted them that they had a problem with their analysis: either with the first (1999) or the second (2001) set of estimations. The hair sample allegedly removed from Grand Duke George offered contradictory results, and did not correlate with the mtDNA profiles of the Grand Duke, analyzed in the United States Armed Forces Institute of Pathology by Professor Ivanov. According to their 2001 results, the only discrepancy was that Nagai and Popov were unable to detect heteroplasmy at position 16169 during the course of their investigations.
Regardless of these intrinsic problems, the two sets of projects had enabled this panel of researchers to achieve their ultimate goal: to presume that the remains found in Ekaterinburg did not belong to the last Russian Imperial Family. Indeed, Mrs. Kulikovskaya, who funded the Nagai/Popov project, expressed that their result “attaches great importance to their identification test.”  However, the broad scientific community remains unaware of these “identification tests.”
Table 3: Summary of mtDNA results obtained by various laboratory investigators
Matrilineal descendants of Louise Hesse-Cassel
Heteroplasmy or fixation at 16169 position on mtDNA sequence
Principle Investigators and year of assessments
Countess Ksenya Sfiris
Gill and Ivanov } 1993
Grand Duke George
Nicholas II [repeat assay]
Ksenya Sfiris [repeat assay]
Ivanov } 1995
Skeleton # 4
[Nicholas II: Repeat assay]
Grand Duke George
Grand Duke George
7 heteroplasmies - none at position 16169
Nagai/Popov } 2001
In 2004, Dr Kevin Sullivan, a member of Dr Gill’s team which analyzed the Imperial remains at the British Forensic Laboratories, stated:
“We published [in Nature Genetics] that the remains were authentic, in that they correlated with the samples of DNA extracted from living relatives of the Imperial Family on the maternal side.”
Regrettably, the lack of
understanding and “reading” DNA profiles triggered confusion among
non-scientific circles. The declaration by Nagai before the local media in
Romanov Family dismisses the Japanese findings.
In a recent interview to Izvestiya in 2005, the Head of the Romanov Family, Prince Nicholas made a number of remarks about the course of recent events. “The final investigations which were headed by the investigator Soloviev were exhausted” and that all the questions, which the Moscow Patriarchy posed, were answered. They continue to accept the forensic conclusions contained in the 1998 Soloviev Commission Report “and all the technical information, which was printed and published. The conclusions were based on proof that was beyond absolute doubt …”  Thus after a few years, only serious concerns must be raised as to why the Japanese expert (whose broad profile included silkworms, hepatitis and epilepsy studies) became involved. Furthermore, Prince Nicholas contended that no one knew of Nagai’s “expertise” - the very same person who used material wwith suspect preservation, with no guarantee that that it is genuine.
The Most Disturbing Controversy of All
During the course of
our investigations we located an interview, which Soloviev gave to Moscovskii Komsomolets in February 2005,
which featured a number of very disturbing pieces of information. This senior
Soloviev was how Dr Nagai received hair samples from Grand Duke George and a
fragment of skeleton # 4 (Nicholas II). There was only one way, he asserted: “ …if they were stolen from the
Even more disturbing
was that there was no formal authorization to have any Russian samples to be
Professor Popov was given the opportunity for his right of reply to Soloviev’s interview on 22 March, 2005. The radio interviewer asked Popov “By what method did the remains come to the Japanese geneticist Tatsuo Nagai…and Soloviev asserts that it was you who handed of the remains?” Popov replied “…why should he want to know about this? Now that the investigation is over…” Popov stated that in spite of the fact that all excess samples were to be returned for re-burial some remnants remained in the “Nicholas II Memorial Foundation.” Popov divulged at the same interview that Soloviev refused to part with any portion of the Ekaterinburg remains, outside the nominated experts. Then he just happened to meet Nagai and they mutually arranged to proceed to test those excess remains.  Not only did Popov indirectly admit that there was a problem of legitimate possession and transfer to another interested party, but that the customary protocol - the “chain of custody” issue observed by accredited laboratories was compromised.
The “chain of
custody” is a standard that becomes important when any sample is assessed to
determine its authenticity and provides key information for other experts in
the field when they read the published results. Popov also openly explained how
he, in Nagai’s presence removed a portion of scotch tape appended to Nicholas
II’s coat that was on exhibit at the
We were initially puzzled about
one fundamental point. Why would Popov make such arrangements with Nagai? It
seems he was aware of Nagai’s results allegedly obtained from testing that
Soloviev’s well-informed public statement that the Imperial samples were “stolen” only reached Russian audiences. During his interview for Strana, Professor Ivanov stated that Popov “just simply handed over this national property” and generously predicted that “…it shall remain on his conscience”.
Clearly, under these circumstances no formal publication in any accredited journal such as Nature in English, or even any Russian publication, could ever be contemplated. It may equally explain why we encountered reluctance to offer any response when one of the authors (Helen Azar) contacted Dr Nagai’s associate to discuss their results.
It does seem surprising that no
foreign press picked up on this very important underlying story, assuming
instead that everything was legitimate. The international media, including
Russian émigré newspapers published in
Controversy within the
these circumstances it is very difficult to comprehend why Nagai was invited,
in December 2004, to announce his results directly to the Moscow Patriarchate
representative, Bishop Alexander Dmitrovskii, to whom he erroneously declared
that those remains belonged not to Nicholas II and his family and
retainers, but to other unknown individuals.
Following that encounter, the
Despite their admission, the
Church preferred to believe this single unknown DNA “expert” from
The Procurator and Commissioner, Soloviev disputed this point, asserting that the Church did indeed receive replies to those 10 questions in the form of a consolidated publication of the Commission’s Final Report titled Pokayanie (Repentance), which specifically addressed all the Church’s concerns, including all letters, which we, the authors, also have in our possession. In fact it was Boris Nemtsov; the President of the Sokolov Commission who personally handed over the Final Report to Patriarch Alexei II.  Furthermore Nemtsov added that the Church never spoke about the Commission’s work – not at their meetings nor at any government meetings.
RECA praised Professor Nagai for his work that disputed the authenticity of Imperial remains. To honor his efforts, they presented him with an icon from the Moscow Patriarch (the first foreigner to receive such a gift) and presented him with an honorable title of some kind. In response Nagai stated: “I am so proud of this as a scientist who solved this mystery.” (Translation courtesy of Mr. Junichi Hayashi).
During our investigation, the authors attempted to clarify
Dr Nagai’s data interpretation directly with him, but learned that he recently
retired from the university and is no longer reachable by email. His associate,
Dr Toshio Okazaki, with whom we
communicated briefly (via email), declined to answer any questions about the
Romanov study, even though he actively participated in it along with Dr Nagai.
He insisted that we speak directly with Dr Nagai, who unfortunately was
The President of the Russian Federation, Sergei Mironov took a peculiar middle road regarding the problems relating to the authenticity of the remains during his visit to the Fortress in 2005 in which he expressed the following opinion: “There are different points of view …It is best to appear that no problems exist.” We believe that this attitude is far from acceptable, and only evades some very serious issues of national importance that need to be addressed.
Dr Alexander Avdonin, one of the principles who facilitated that the Imperial remains be raised to the surface some fifteen years ago and now heads Obretenye Foundation believes that:
“We are dealing with single-minded company (RECA) who under the guise of establishing historic truth … discredit the names of well known Russian politicians, entire government institutions of Russia - the President … the General Procurator, and lead the people of a great nation astray.” 
He asks: “Interestingly, what kind of a reaction would those Japanese express if their museums were plundered of relics, and from their sacred graves the remains of their great compatriots were pilfered?”
Obretenye is waiting for the Russian
people to “actively react” against
those who want to damage not just
Our investigation has unexpectedly identified an aspect of scientific research at its most distasteful level. We can only hope that these revelations will at least help you, the reader, understand that not all scientific endeavors advance medical science, nor in this case confirm historic truth.
To conclude our discussion on a more positive note, it is rather poignant that Tihon Kulikovsky did in fact help authenticate the Ekaterinburg remains in the very end, despite his earlier misgivings in doing so.
(Photo by Margarita Nelipa)
The authors wish to extend their gratitude to Peter Sarandinaki, President and founder of the S.E.A.R.C.H. Foundation http://www.searchfoundationinc.org/index.html for his generous support and warm friendship.
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