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Concept: Mummy

484

Egypt, located on the isthmus of Africa, is an ideal region to study historical population dynamics due to its geographic location and documented interactions with ancient civilizations in Africa, Asia and Europe. Particularly, in the first millennium BCE Egypt endured foreign domination leading to growing numbers of foreigners living within its borders possibly contributing genetically to the local population. Here we present 90 mitochondrial genomes as well as genome-wide data sets from three individuals obtained from Egyptian mummies. The samples recovered from Middle Egypt span around 1,300 years of ancient Egyptian history from the New Kingdom to the Roman Period. Our analyses reveal that ancient Egyptians shared more ancestry with Near Easterners than present-day Egyptians, who received additional sub-Saharan admixture in more recent times. This analysis establishes ancient Egyptian mummies as a genetic source to study ancient human history and offers the perspective of deciphering Egypt’s past at a genome-wide level.

Concepts: Africa, North Africa, Ancient Egypt, Egypt, Slavery, Ancient history, Mummy, Nubia

34

In an effort to better understand the ancestral state of the human distal gut microbiome, we examine feces retrieved from archaeological contexts (coprolites). To accomplish this, we pyrosequenced the 16S rDNA V3 region from duplicate coprolite samples recovered from three archaeological sites, each representing a different depositional environment: Hinds Cave (∼8000 years B.P.) in the southern United States, Caserones (1600 years B.P.) in northern Chile, and Rio Zape in northern Mexico (1400 years B.P.). Clustering algorithms grouped samples from the same site. Phyletic representation was more similar within sites than between them. A Bayesian approach to source-tracking was used to compare the coprolite data to published data from known sources that include, soil, compost, human gut from rural African children, human gut, oral and skin from US cosmopolitan adults and non-human primate gut. The data from the Hinds Cave samples largely represented unknown sources. The Caserones samples, retrieved directly from natural mummies, matched compost in high proportion. A substantial and robust proportion of Rio Zape data was predicted to match the gut microbiome found in traditional rural communities, with more minor matches to other sources. One of the Rio Zape samples had taxonomic representation consistent with a child. To provide an idealized scenario for sample preservation, we also applied source tracking to previously published data for Ötzi the Iceman and a soldier frozen for 93 years on a glacier. Overall these studies reveal that human microbiome data has been preserved in some coprolites, and these preserved human microbiomes match more closely to those from the rural communities than to those from cosmopolitan communities. These results suggest that the modern cosmopolitan lifestyle resulted in a dramatic change to the human gut microbiome.

Concepts: Human, Gut flora, Primate, Mexico, Match, Archaeology, Mummy, Las Vegas Strip

32

We applied, for the first time, next-generation sequencing (NGS) technology on Egyptian mummies. Seven NGS datasets obtained from five randomly selected Third Intermediate to Graeco-Roman Egyptian mummies (806 BC-124AD) and two unearthed pre-contact Bolivian lowland skeletons were generated and characterised. The datasets were contrasted to three recently published NGS datasets obtained from cold-climate regions, i.e. the Saqqaq, the Denisova hominid and the Alpine Iceman. Analysis was done using one million reads of each newly generated or published dataset. Blastn and megablast results were analysed using MEGAN software. Distinct NGS results were replicated by specific and sensitive polymerase chain reaction (PCR) protocols in ancient DNA dedicated laboratories. Here, we provide unambiguous identification of authentic DNA in Egyptian mummies. The NGS datasets showed variable contents of endogenous DNA harboured in tissues. Three of five mummies displayed a human DNA proportion comparable to the human read count of the Saqqaq permafrost-preserved specimen. Furthermore, a metagenomic signature unique to mummies was displayed. By applying a “bacterial fingerprint”, discrimination among mummies and other remains from warm areas outside Egypt was possible. Due to the absence of an adequate environment monitoring, a bacterial bloom was identified when analysing different biopsies from the same mummies taken after a lapse of time of 1.5 years. Plant kingdom representation in all mummy datasets was unique and could be partially associated with their use in embalming materials. Finally, NGS data showed the presence of Plasmodium falciparum and Toxoplasma gondii DNA sequences, indicating malaria and toxoplasmosis in these mummies. We demonstrate that endogenous ancient DNA can be extracted from mummies and serve as a proper template for the NGS technique, thus, opening new pathways of investigation for future genome sequencing of ancient Egyptian individuals.

Concepts: DNA, Polymerase chain reaction, Molecular biology, Apicomplexa, DNA sequencing, Toxoplasmosis, Ancient Egypt, Mummy

18

The mummies of Kha and his wife Merit were found intact in an undisturbed tomb in western Thebes near the ancient workers' village of Deir el-Medina. Previous MDCT (this abbreviation needs spelling out) investigations showed that the bodies of Kha and Merit did not undergo classical royal 18th Dynasty artificial mummification, which included removal of the internal organs. It was, therefore, concluded that the retention of the viscera in the body, combined with an absence of canopic jars in the burial chamber, meant the couple underwent a short and shoddy funerary procedure, despite their relative wealth at death. Nevertheless, all internal organs - brain, ocular bulbs/ocular nerves, thoracic and abdominal organs - showed a very good state of preservation, which contradicts the previous interpretation above. In order to better understand the type of mummification used to embalm these bodies, both wrapped mummies were reinvestigated using new generation X-ray imaging and chemical microanalyses Here we provide evidence that both individuals underwent a relatively high quality of mummification, fundamentally contradicting previous understanding. Elucidated “recipes”, whose components had anti-bacterial and anti-insecticidal properties, were used to treat their bodies. The time and effort undoubtedly employed to embalm both Kha and Merit and the use of imported costly resins, notably Pistacia, do not support the previously held view that the two individuals were poorly mummified. Despite a lack of evisceration, the approach clearly allowed their in situ preservation as well as affording a fairly successful mummification.

Concepts: X-ray, Heart, Ancient Egypt, Organs, Embalming, Wealth, Burial, Mummy

18

What originally appeared to be only an external cast of an anuran ‘mummy’ from the Quercy Phosphorites (southwestern France) was described as Rana plicata during the 19th century. Its geographical provenance is only vaguely known; therefore its precise age within the Paleogene was uncertain. The taxon was erected on the basis of the external morphology of the specimen, which includes few diagnostic characters. As a further complication, the name Rana plicata was recently shown to be unavailable at the time of the description, and the name Rana cadurcorum was proposed as a replacement. In order to see whether internal features were fossilized, the fossil was CT scanned. This showed that a large part of the skeleton is preserved. Unexpectedly, the scans revealed that the skull of the mummy is almost identical to that of Thaumastosaurus gezei, another anuran from the late middle or late Eocene of the Quercy Phosphorites. The few observed differences are attributable to intraspecific and ontogenetic variation, and R. cadurcorum is a junior subjective synonym of T. gezei. The mummy is therefore probably from the same time interval as T. gezei. The latter was previously known only by its skull, but the mummy provides important information on the postcranial skeleton. Earlier assessments, based only on the skull, placed Thaumastosaurus close to South American hyloid anurans, but a new phylogenetic analysis including postcranial characters reveals ranoid affinities. This study exemplifies the usefulness of modern imaging technologies that allow non-destructive study of previously inaccessible internal anatomical features.

Concepts: Biology, Tetrapod, Fossil, 19th century, Oligocene, Mummy, The Mummy, Imhotep

12

The paleopathological, paleoradiological, histological, molecular and forensic investigation of a female mummy (radiocarbon dated 1451-1642 AD) provides circumstantial evidence for massive skull trauma affecting a young adult female individual shortly before death along with chronic infection by Trypanosoma cruzi (Chagas disease). The mummy (initially assumed to be a German bog body) was localized by stable isotope analysis to South America at/near the Peruvian/Northern Chilean coast line. This is further supported by New World camelid fibers attached to her plaits, typical Inca-type skull deformation and the type of Wormian bone at her occiput. Despite an only small transverse wound of the supraorbital region computed tomography scans show an almost complete destruction of face and frontal skull bones with terrace-like margins, but without evidence for tissue reaction. The type of destruction indicates massive blunt force applied to the center of the face. Stable isotope analysis indicates South American origin: Nitrogen and hydrogen isotope patterns indicate an extraordinarily high marine diet along with C4-plant alimentation which fits best to the coastal area of Pacific South America. A hair strand over the last ten months of her life indicates a shift to a more “terrestric” nutrition pattern suggesting either a move from the coast or a change in her nutrition. Paleoradiology further shows extensive hypertrophy of the heart muscle and a distended large bowel/rectum. Histologically, in the rectum wall massive fibrosis alternates with residual smooth muscle. The latter contains multiple inclusions of small intracellular parasites as confirmed by immunohistochemical and molecular ancient DNA analysis to represent a chronic Trypanosoma cruzi infection. This case shows a unique paleopathological setting with massive blunt force trauma to the skull nurturing the hypothesis of a ritual homicide as previously described in South American mummies in an individual that suffered from severe chronic Chagas disease.

Concepts: Bone, Infectious disease, Chagas disease, Skull, Trypanosoma, South America, Trypanosoma cruzi, Mummy

10

The funeral preparations for ancient Egyptian dead were extensive. Tomb walls were often elaborately painted and inscribed with scenes and objects deemed desirable for the afterlife. Votive objects, furniture, clothing, jewelry, and importantly, food including bread, cereals, fruit, jars of wine, beer, oil, meat, and poultry were included in the burial goods. An intriguing feature of the meat and poultry produced for the deceased from the highest levels of Egyptian society was that they were mummified to ensure their preservation. However, little is known about the way they were prepared, such as whether balms were used, and if they were used, how they compared with those applied to human and animal mummies? We present herein the results of lipid biomarker and stable carbon isotope investigations of tissues, bandaging, and organic balms associated with a variety of meat mummies that reveal that treatments ranged from simple desiccation and wrapping in bandages to, in the case of the tomb of Yuya and Tjuia (18th Dynasty, 1386-1349 BC), a balm associated with a beef rib mummy containing a high abundance of Pistacia resin and, thus, more sophisticated than the balms found on many contemporaneous human mummies.

Concepts: Death, Carbon, Ancient Egypt, Embalming, Mummy, British Museum

7

Many studies have been concerned with the ancient Egyptian mummification method; nevertheless, little effort has been made to explore it experimentally. The goal of this study is to apply evidence-based diagnostic criteria and state-of-the art methodology in order to improve knowledge on soft tissues preservation and postmortem alterations. Two human lower limbs (LL) from a female donor were (1) “naturally” mummified by dry heat and (2) artificially in natron. At specific time intervals a macroscopic and radiological examination of the LL was performed and skin and muscle samples were taken for histological and biomolecular analysis. Temperature, humidity, pH, and weight of the LL were systematically measured. The mummification by dry heat was stopped after 7 days due to unexpected lack of mummification progress. The mummification in natron was completed successfully after 208 days. The humidity, the external temperature, and the pH were proven with Pearson correlation and principal component analysis as important factors for the mummification process. The steady removal of water from the tissues through the natron has prevented the putrefaction. This is also evident in the absence of bacteria or fungi through the microbiological analysis. The histological analysis revealed very good preservation of the skin and the muscle tissues. In the muscular sample certain degree of structural disintegration can be seen, particularly affecting the epimysium whilst in the skin samples the epidermis, especially the stratum corneum, is mostly affected. The samples show better preservation compared with ancient Egyptian sections and other mummified tissues from historic or forensic context. Anat Rec, 298:974-987, 2015. © 2015 Wiley Periodicals, Inc.

Concepts: Energy, Muscle, Tissues, Skin, Tissue, Karl Pearson, Ancient Egypt, Mummy

5

To correlate the radiologic findings detected with computed tomography scan with anthropological data in 13 naturally mummified bodies discovered during works of recovery of an ancient church in a crypt in Roccapelago, in the Italian Apennines.

Concepts: Medical imaging, Tomographic reconstruction, Mummy

4

The analysis of nucleic acids in ancient samples is largely limited to DNA. Small non-coding RNAs (microRNAs) are known to be evolutionary conserved and stable. To gain knowledge on miRNAs measured from ancient samples we profiled microRNAs in cryoconserved mummies. First, we established the approach on a World War One warrior, the “Kaiserjäger”, which has been preserved for almost one century. Then, we profiled seven ancient tissue specimens including skeletal muscle, stomach mucosa, stomach content and two corpus organ tissues of the 5,300 year-old copper age mummy Iceman and compared these profiles to the presence of organ-specific miRNAs in modern tissues. Our analyses suggest the presence of specific miRNAs in the different Iceman’s tissues. Of 1,066 analyzed human miRNAs 31 were discovered across all biopsies and 87 miRNAs were detected only in a single sample. To check for potential microbiological contaminations, all miRNAs detected in Iceman samples and not present in ancient samples were mapped to 14,582 bacterial and viral genomes. We detected few hits (3.9% of miRNAs compared to 3.6% of miRNAs). Interestingly, the miRNAs with higher abundance across all ancient tissues were significantly enriched for Guanine (p-value of 10-13) and Cytosine (p-value of 10-7). The same pattern was observed for modern tissues. Comparing miRNAs measured from ancient organs to modern tissue patterns highlighted significant similarities, e.g. for miRNAs present in the muscle. Our first comprehensive analysis of microRNAs in ancient human tissues indicates that these stable molecules can be detected in tissue specimens after 5,300 years. .

Concepts: DNA, Gene, Virus, RNA, Organ, Nucleic acid, Guanine, Mummy