Molecular Exploration of the First-Century Tomb of the Shroud in Akeldama, Jerusalem

May combine two claims.

Textiles from a burial shroud, hair and skeletal remains were found in the tomb.

“More than twenty stone ossuaries for collecting human bones were found, along with textiles from a burial shroud, hair and skeletal remains.”

Probably not a substantial claim: may not pass S3 (load-bearing).

More than twenty stone ossuaries for collecting human bones were found in the tomb.

“More than twenty stone ossuaries for the collection of human bone were found”

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

The tomb had been illegally entered and looted before archaeologists reached the site.

“The tomb had been entered and looted before archaeologists reached the site.”

Probably not a substantial claim: may not pass S4 (based).

Sealing a loculus with white plaster is quite rare in the first-century tombs studied around Jerusalem.

“a practice which is quite rare in the first century tombs studied around Jerusalem.”

May combine two claims.

The calibrated calendar date range of the shroud textile is 50 BCE-16 CE at one sigma and 95 BCE-53 CE at two sigma.

“with calibrated calendar date ranges of one sigma: 50 BCE -16 CE and two sigma: 95 BCE -53 CE.”

May combine two claims.

The male individual suffered from tuberculosis and leprosy.

“that suffered from tuberculosis and leprosy”

May combine two claims.

Neither contamination nor the archaeologist's DNA was found in any of the results.

“however neither contamination nor the archaeologist's DNA was found in any of the results.”

May combine two claims.

The mtDNA analysis identified two mtDNA profiles shared between two individuals and one mtDNA profile shared between three individuals.

“This mtDNA analysis has identified two mtDNA profiles that are shared between two individuals and one mtDNA profile that is shared between three individuals”

May combine two claims.

The female and one infant sample from SC2 had mtDNA type 2, and the second infant had mtDNA type 3.

“The female and one of the infant samples had mtDNA type 2, and the second infant had mtDNA type 3.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

The fourth sample tested from SC7 did not produce a reliable mtDNA sequence but produced an IS6110 sequence and was not included in the study thereafter.

“The fourth sample tested from SC7 did not produce a reliable mtDNA sequence but produced an IS6110 sequence and was not included in the study thereafter.”

May combine two claims.

Four samples tested from SC7 were identified as one male and one female with different mtDNA types (type 5 and type 4 respectively).

“There were four samples tested from the SC7 location (Figure 1). These were identified as one male and one female with different mtDNA types (mtDNA type 5 and type 4 respectively).”

May combine two claims.

The single sample tested from SC9 was identified as a male with mtDNA type 6, which did not match any other member of the tomb that was tested.

“There was only one sample tested from the SC9 location (Figure 1). This sample was identified as a male with mtDNA type 6, which did not match any other member of the tomb that was tested.”

May combine two claims.

The skeletal remains from within the plaster-sealed parts of the tomb (SC1 and SC2) are positive for M. tuberculosis and one of them for both M. tuberculosis and M. leprae.

“It is interesting to note that the skeletal remains from within the plaster-sealed parts of the tomb (SC1 and SC2) are positive for M. tuberculosis and one of them for both M. tuberculosis and M. leprae.”

May combine two claims.

The two SC17 samples had different mtDNA types, the female mtDNA type 5 and the other mtDNA type 7.

“These samples both had different mtDNA types, the female mtDNA type 5 and the other mtDNA type 7.”

May combine two claims.

Two samples analyzed from SC4 produced the same mtDNA type 4 and were both male.

“There were two samples analyzed from the Tomb of the Shroud SC4 location (Figure 1). Both these samples produced the same mtDNA type 4 and were both found to be male.”

May combine two claims.

There were two maternally related pairs of individuals: adult female SC2 and infant SC2, and adult female SC17 and adult male SC7.

“From this analysis there have been identified two maternally unrelated male individuals (both adults from SC1 and SC9), two maternally unrelated sex indeterminate individuals (one adult SC17 and one infant SC2) and two maternally related pairs of individuals (adult female SC2 and infant SC2 and adult female SC17 and adult male SC7).”

May combine two claims.

Four samples were tested from SC2; they were identified as a female and two infants for which sex was not identified.

“There were four samples tested from the Tomb of the Shroud SC2 location (i.e. the ossilegium niche in the north wall of the lower chamber) (Figure 1). These were identified as a female and two infants for which their sex was not identified.”

May combine two claims.

All amplifications obtained from archaeological samples were identical to the NCBI Genbank sequence for IS6110 (X17348, bases 788-863) and the S12 protein (X70995).

“All amplifications obtained from archaeological samples were identical to the NCBI Genbank sequence for IS6110 (sequence X17348, bases 788–863) and the S12 protein (sequence X70995 in the NCBI database) (Table 6 and 7).”

Classified as probably not a substantial claim. May combine two claims.

Researchers adhered to standard ancient DNA and forensic protocols including laboratory coats, sleeves, double gloving, face mask, and frequent glove changes.

“The researchers adhered to all standard ancient DNA and forensic protocols including the use of laboratory coats, sleeves, double gloving and face mask with frequent changes of gloves.”

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

The Tomb of the Shroud is one of very few examples of a preserved shrouded human burial.

“The Tomb of the Shroud is one of very few examples of a preserved shrouded human burial”

May combine two claims.

Mitochondria are maternally inherited organelles suitable for characterization of maternal relationships.

“Mitochondria are maternally inherited organelles, suitable for characterization of maternal relationships [11], [12]”

• cites [11] Bini C Pappalardo G 2005 mtDNA HVI length heteroplasmic profile in different tissues of maternally related members. Forensic Science International 152 35 38 15939174 sourceNot checked — no full text of the source
• cites [12] Schultes T Hummel S Herrmann B 2000 Ancient DNA-typing approaches for the determination of kinship in a disturbed collective burial site. Anthropol Anz 58 37 44 10816784 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

The first report of M. leprae DNA in human remains was from the seventh century C.E.

“This was quickly followed by the first report of M. leprae DNA in human remains from the seventh century C.E. [17], [18].”

• cites [17] Rafi A Spigelman M Stanford J Lemma E Donoghue H 1994 Mycobacterium leprae DNA from ancient bone detected by PCR. Lancet 343 1360 1361 7910345 sourceNot checked — no full text of the source
• cites [18] Rafi A Spigelman M Stanford J Lemma E Donoghue H 1994 DNA of Mycobacterium leprae Detected by PCR in Ancient Bone. International journal of Osteoarchaeology 4 287 290Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

Probably not a substantial claim: may not pass S3 (load-bearing).

A separate room with a different set of pipettes was used for handling PCR product.

“A separate room with a different set of pipettes was used for handling PCR product.”

Classified as probably not a substantial claim.

Bone samples were UV irradiated at 254 nm for 24-48 hours to crosslink surface contaminating DNA from soil bacteria or handling.

“These were first placed under UV irradiation with 254 nm wavelength for 24–48 hours, depending on the size of the sample, to crosslink any surface contaminating DNA from soil bacteria or handling.”

The quote only partly states this claim.

The Tomb of the Shroud is the only example of a plaster sealed loculus with remains genetically confirmed to have belonged to a shrouded male individual.

“the only example of a plaster sealed loculus with remains genetically confirmed to have belonged to a shrouded male individual that suffered from tuberculosis and leprosy dating to the first-century C.E.”

May combine two claims.

The target mtDNA region is small and thus errors could be introduced in the identification of relationship amongst a random sample population.

“However the target region is small and thus errors could be introduced in the identification of relationship amongst a random sample population.”

Probably not a substantial claim: may not pass S3 (load-bearing).

The Paleo-DNA Laboratory follows strict standard operating procedures (SOPs).

“The Paleo-DNA Laboratory also carries out forensic case work and follows strict standard operating procedures (SOPs).”

May combine two claims.

Identification of the causative agent for the lesion is hindered by the disturbed nature of the skeletal remains and limited preservation of skeletal elements.

“The identification of the causative agent for the lesion is hindered by the disturbed nature of the skeletal remains (caused by the illegal looters of the tomb), and limited preservation of skeletal elements, especially those in association with the affected region.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Positive control DNA was diluted to simulate ancient DNA concentrations and added in a separate room from PCR preparation.

“The positive control DNA was diluted to simulate ancient DNA concentrations and was added to the positive control reaction tubes in a separate room from where the PCR reactions were prepared, before being loaded onto the thermal cycler.”

Classified as probably not a substantial claim.

Extraction negatives were used to ensure no contamination entered the samples.

“The whole procedure was performed with extraction negatives to ensure that no contamination had entered the samples during this process.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Extraction negative control tubes were always processed in parallel with tubes containing samples.

“Extraction negative control tubes were always processed in parallel with tubes containing samples.”

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based). May combine two claims.

The discovery of preserved shroud textile in this tomb is rare because high humidity in the Jerusalem area does not usually allow preservation of organic material.

“This discovery is rare because high levels of humidity in the Jerusalem area does not usually allow for the preservation of organic material.”

• AssumptionRests on an assumption: High levels of humidity in the Jerusalem area do not usually allow for the preservation of organic material.

Probably not a substantial claim: may not pass S3 (load-bearing).

The first pathogenic DNA recovered from archaeological remains was M. tuberculosis.

“The first pathogenic DNA recovered from archaeological remains was M. tuberculosis [16].”

• cites [16] Spigelman M Lemma E 1993 The use of the polymerase chain reaction (PCR) to detect Mycobacterium tuberculosis in ancient skeletons. International Journal of Osteoarchaeology 3 137 143Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Laboratory controls are taken every month, with surfaces swabbed and analyzed for contamination.

“Laboratory controls are taken every month, where all the surfaces are swabbed and analyzed for contamination.”

Classified as probably not a substantial claim. May combine two claims. The quote only partly states this claim.

The working area and equipment were bleached, washed, sterilized, and UV-irradiated before and after each sample preparation.

“The working area was bleached and washed with 3.0% (w/v) sodium hypochlorite solution followed by 70% ethanol before and after each sample preparation and all equipment was sterilized before use. The equipment and the work area were UV-irradiated prior to each preparation.”

Classified as probably not a substantial claim. May combine two claims.

Stringent precautions were taken to avoid contamination in London, including sterile reagents, tubes, plugged tips, clean protective clothing, and frequent glove changes.

“Stringent precautions were taken to avoid contamination. Sterile reagents, tubes and plugged tips were used and clean protective clothing worn, with frequent glove changes.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Co-infection of two mycobacterial diseases is not uncommon and has been previously identified.

“however, the co-infection of two mycobacterial diseases is not uncommon and has been previously identified [27].”

• cites [27] Donoghue HD Marcsik A Matheson C Vernon K Nuorala E 2005 Co-infection of Mycobacterium tuberculosis and Mycobacterium leprae in human archaeological samples: a possible explanation for the historical decline of leprosy. Proceedings of the Royal Society of London Series B Biological Sciences 272 389 394 15734693 PMC1634979 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

The Paleo-DNA Laboratory has separated pre-PCR, post-PCR and PCR areas all with separate air handling systems.

“The facility has separated pre-PCR, post-PCR and PCR areas all with separate air handling systems.”

Probably not a substantial claim: may not pass S3 (load-bearing).

The preservation and persistence of ancient bacterial DNA in the archaeological record is controversial.

“The preservation and persistence of ancient bacterial DNA in the archaeological record is controversial [25]”

• cites [25] Willerslev E Hansen AJ Ronn R Brand TB Barnes I 2004 Long-term persistence of bacterial DNA. Curr Biol 14 R9 10 14711425 sourceNot checked — no full text of the source

Classified as probably not a substantial claim.

Standard molecular archaeology protocols were applied for analysis of the Tomb of the Shroud samples at the Hebrew University and replicated at Lakehead University and University College London.

“Standard molecular archaeology protocols were applied for analysis of the Tomb of the Shroud samples at the ancient DNA laboratory in the Kuvin Centre for Tropical and Infectious Diseases, Hebrew University (Israel) and replicated at the Paleo-DNA Laboratory, Lakehead University (Canada) and at the Department of Infection, University College London (UK).”

Probably not a substantial claim: may not pass S3 (load-bearing). The quote only partly states this claim.

Both negative and positive controls were always included in PCR.

“The DNA extract (10.0 µl) was added to each tube and both negative and positive controls were always included.”

Classified as probably not a substantial claim.

Bone material from inside the bone was removed by a dental pick while the surface was left intact to ensure no contamination.

“To ensure there was no contamination, bone material from inside the bone was removed by a dental pick while the surface of the bone was left intact.”

Classified as probably not a substantial claim.

The archaeologist's DNA was characterised for reference to eliminate possible contamination.

“The archaeologist's DNA was characterised for reference to eliminate possible contamination”

Probably not a substantial claim: may not pass S3 (load-bearing).

Permission for this research was provided by the Israeli Antiquities Authority and the archaeologists who excavated the material.

“Permission for this research was provided by the Israeli Antiquities Authority and the archaeologists who excavated the material.”

Classified as probably not a substantial claim.

Multiple primer sets were included to address fragmentation.

“These multiple primer sets were included to address fragmentation.”

Classified as probably not a substantial claim.

PCR reagent preparations were physically separated from sample preparation and PCR in separate rooms with separate air supply.

“Standard precautions to minimize contamination were employed, sterile tubes and plugged tips under UV hoods where PCR reagent preparations were physically separated, from both sample preparation and PCR, in separate rooms with separate air supply.”

Classified as probably not a substantial claim.

The Paleo-DNA Laboratory is a dedicated ancient DNA facility with stringent precautions and protocols for degraded DNA.

“The Paleo-DNA Laboratory is a dedicated ancient DNA facility with stringent precautions and protocols employed for the analysis of degraded DNA.”

Probably not a substantial claim: may not pass S3 (load-bearing).

The recovery of ancient or degraded DNA is challenging and hindered by chemical modifications and fragmentation.

“The recovery of ancient or degraded DNA is challenging and hindered by forms of damage, including chemical modifications and fragmentation of the DNA [3], [4], [5].”

• cites [3] O'Rourke DH Hayes MG Carlyle SW 2000 Ancient DNA Studies in Physical Anthropology. Annual Review of Anthropology 29 217 242Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

• cites [4] Willerslev E Cooper A 2005 Ancient DNA. Proc Biol Sci 272 3 16 15875564 PMC1634942 sourceNot checked — no full text of the source
• cites [5] Hofreiter M Serre D Poinar HN Kuch M Pääbo S 2001 Ancient DNA. Nature Reviews Genetics 2 353 359 11331901 sourceNot checked — no full text of the source

Classified as probably not a substantial claim.

Sediment samples from around the tomb and scrapings from the rock walls and loculus roof were analyzed for pathogenic mycobacterial DNA or environmental DNA causing non-specific results.

“Sediment samples from around the tomb and scrapings from the rock walls and loculus roof were analyzed for the presence of any pathogenic mycobacterial DNA or environmental DNA causing non-specific results.”

Probably not a substantial claim: may not pass S3 (load-bearing).

The M. tuberculosis and M. leprae procedure was replicated in London on samples of the SC1 and SC2 individuals.

“The amplification of the M. tuberculosis and M. leprae procedure was replicated in London on samples of the SC1 and SC2 individuals.”

May combine two claims.

Amplification of mitochondrial DNA from these same samples and both positive and negative results for pathogenic DNA from these tombs, in addition to logical and expected results for the haplogroups identified, supports authentic results.

“Amplification of the mitochondrial DNA from these same samples and both positive and negative results for pathogenic DNA from these tombs in addition to logical and expected results for the haplogroups identified supports authentic results.”

May combine two claims.

A co-infection of tuberculosis and leprosy was an unexpected discovery in the remains of the shrouded individual and presents a significant and unique anthropological example of contagious disease in the past.

“A co-infection of tuberculosis and leprosy was an unexpected discovery in the remains of the shrouded individual and present a significant and unique anthropological example of contagious disease in the past.”

May combine two claims.

The skeletal remains in the wall niche were those of the two infants with tuberculosis and a maternally related adult female, possibly the mother.

“The skeletal remains in the wall niche were those of the two infants with tuberculosis and a maternally related adult female, possibly the mother.”

May combine two claims.

Identification of M. tuberculosis from the tooth pulp of these individuals suggests that this pathogen was in the bloodstream and therefore throughout the body as miliary tuberculosis.

“Identification of M. tuberculosis from the tooth pulp of these individuals suggests that this pathogen was in the bloodstream and therefore throughout the body as miliary tuberculosis and all three individuals would have almost certainly have died from a disseminated bacterial infection.”

May combine two claims.

The co-infection of leprosy with tuberculosis is often the cause of death as M. leprae infection generates a compromised immune system, vulnerable to a subsequent M. tuberculosis infection that is still fatal in some parts of the world today.

“The co-infection of leprosy with tuberculosis is often the cause of death as M. leprae infection generates a compromised immune system, vulnerable to a subsequent M. tuberculosis infection that is still fatal in some parts of the world today.”

• AssumptionRests on an assumption: The presence of both M. tuberculosis and M. leprae DNA is associated with immuno-suppressed individuals, which is a feature of the multi-bacillary or lepromatous form of leprosy.
• AssumptionRests on an assumption: On infection the individual becomes immuno-compromised and the chance of co-infection by M. tuberculosis and other pathogens occurs.

May combine two claims.

The molecular identification of tuberculosis in the Jerusalem tomb is a significant contribution to the archaeological interpretation of this site and suggests a reason for the plaster sealing of the loculus containing the shrouded man and for the additional individuals sealed in the wall niche.

“The molecular identification of tuberculosis in the Jerusalem tomb is a significant contribution to the archaeological interpretation of this site and suggests a reason for the plaster sealing of the loculus containing the shrouded man and for the additional individuals sealed in the wall niche.”

Probably not a substantial claim: may not pass S3 (load-bearing).

A secondary co-infection of tuberculosis in Hansen's patients is found in 25% of modern cases.

“A secondary co-infection of tuberculosis, in Hansen's patients is found in 25% of modern cases [50], and up to one-third of the world's population is currently infected with tuberculosis (WHO 2007: World Health Organisation: http://www.who.int/tb).”

• cites [50] Jayalakshmi P Looi LM Lim KJ Rajogopalan K 1987 Autopsy findings in 35 cases of leprosy in Malaysia. Int J Lepr Other Mycobact Dis 55 510 514 3655465Not checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

Up to one-third of the world's population is currently infected with tuberculosis.

“A secondary co-infection of tuberculosis, in Hansen's patients is found in 25% of modern cases [50], and up to one-third of the world's population is currently infected with tuberculosis (WHO 2007: World Health Organisation: http://www.who.int/tb).”

• cites WHO 2007: World Health Organisation: http://www.who.int/tbNot matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

Probably not a substantial claim: may not pass S3 (load-bearing).

Molecular pathology adds a new dimension to the archaeological exploration of disease in ancient times, especially in regard to the remains of infants where there is an absence of bone pathologies.

“Molecular pathology adds a new dimension to the archaeological exploration of disease in ancient times, especially in regard to the remains of infants where there is an absence of bone pathologies.”

May combine two claims.

The maternal relatedness of these individuals supports inferences that this Akeldama site was a family tomb which was in use in the first-century C.E.

“The maternal relatedness of these individuals supports inferences that this Akeldama site was a family tomb which was in use in the first-century C.E.”

May combine two claims.

The presence of the lesion supports the genetic identification of M. tuberculosis bacilli in all the skeletal elements analyzed and the severe nature of the infection.

“The presence of this lesion supports the genetic identification of M. tuberculosis bacilli in all the skeletal elements analyzed and the severe nature of the infection.”

May combine two claims.

The mtDNA analysis confirmed the textile was associated with the hair found in loculus SC1 and that the textile was draped or shrouded over the hair and head region.

“The mtDNA analysis also confirmed the textile was associated with the hair found in loculus SC1 and that the textile was draped or shrouded over the hair and head region.”

May combine two claims.

The fourth sample from SC2 had the same mtDNA type and morphological size as one of the three individuals and was concluded to be a replicate sample.

“The fourth sample was found to have the same mtDNA type and morphological size as one of the three individuals concluded to be a replicate sample.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Co-infection presents a plausible explanation for the unusual and extreme lengths people went to in order to contain what was recognized as a seriously debilitating affliction.

“Co-infection present a plausible explanation for the unusual and extreme lengths people went to in order to contain what was recognized as a seriously debilitating affliction.”

May combine two claims.

Genetic analysis confirmed the archaeologist's interpretation of this tomb as a family tomb containing a number of generations and provided important information on the bioarchaeology not available through skeletal analysis.

“Genetic analysis confirmed the archaeologist's interpretation of this tomb as a family tomb containing a number of generations and provided important information on the bioarchaeology not available through skeletal analysis.”

May combine two claims.

It confirms the presence of tuberculosis using molecular methods and is the oldest molecular confirmation of both M. tuberculosis and M. leprae in Jerusalem.

“It confirms the presence of tuberculosis using molecular methods and is the oldest molecular confirmation of both M. tuberculosis and M. leprae in Jerusalem.”

Probably not a substantial claim: may not pass S4 (based).

The individuals from the wall niche undoubtedly underwent secondary burial, meaning that their bones were extracted from a loculus and placed into the niche.

“However, as previously mentioned, the individuals from the wall niche undoubtedly underwent secondary burial, meaning that their bones were extracted from a loculus and placed into the niche.”

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based). May combine two claims.

The prevalence of such a highly contagious disease, particularly for immuno-compromised individuals with leprosy, is not unexpected with inadequate sanitation and demonstrates the significant impact social diseases such as tuberculosis had on society from the low socioeconomic groups up to the more affluent families, such as Tomb the Shroud in first-century Jerusalem.

“The prevalence of such a highly contagious disease, particularly for immuno-compromised individuals with leprosy is not unexpected with inadequate sanitation and demonstrates the significant impact social diseases such as tuberculosis had on society from the low socioeconomic groups up to the more affluent families, such as Tomb the Shroud in first-century Jerusalem.”