Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
2000-03-07
2001-12-11
Arthur, Lisa B. (Department: 1655)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C433S102000, C433S224000, C433S165000
Reexamination Certificate
active
06329149
ABSTRACT:
BACKGROUND OF THE INVENTION
Teeth, as the most durable tissue in the human body, are often all that remain of direct evidence for human occupation of an archaeological site. Dental remains are therefore prized by investigators from numerous disciplines, including physical anthropologists. Dental anthropologists assess teeth for morphological variants that characterize extinct as well as those by which extant populations can be identified (Scott and Turner, 1997).
Through such studies, connections between early hominids and extant primates with current human populations have also been made (Irish, 1998). Through analyses of dental use wear patterns, paleoecologists are able to reconstruct ancient environments. In so doing, new perspectives regarding human physiological as well as cultural adaptations in space and time can be gleaned (Walker, 1976; Grine and Kay, 1988). Depositional differences among skeletal and dental remains enable taphonomists to recreate early hominid paleo-environments (Behrensmeyer, 1975).
Through the assessment of dental stigmata, paleopathologists are able to identify diseases like congenital syphilis, and the existence of nutritional stressors among and between members of mortuary populations (Jacobi, et al., 1992; Katzenberg, 1993; Hillson, 1996; Scott and Turner, 1997; Langsjoen, 1998). Culturally motivated dental alterations (ogsley and Bellande, 1982; Scott and Turner, 1997; Langsjoen, 1998) in addition to environmentally associated occlusal and interproximal wear (Brace, 1975; Blakely and Beck, 1984; Bullington, 1991; Ungar and Spencer, 1999), are also discernable through assessment of dental remains.
This current study was undertaken ancillary to a multi-disciplinary project under the auspices of the Chinese Institute of Archaeology, Chinese Academy of Social Sciences, and the Archaeometry Laboratory at the University of Minnesota, Duluth. The protocol described herein was devised to provide a means by which molecular investigations—e.g. mtDNA haplotypes from the Shang Dynasty Heiheru Site at Anyang, China—could proceed without compromising the integrity of morphologically informative dentition.
Molecular Investigations of Genetic Composition in China
To the extent that advances in the burgeoning field of molecular archaeology have engendered study of DNA from human remains, ancient DNA provides an added dimension to these investigations (Hagelberg, 1994). To this end, mitochondrial DNA (mtDNA) investigations (Cavalli-Sforza et al., 1967, 1988, 1994, 1998) have bolstered the impact of small migration events, such as may occur during trade on otherwise stable gene pools. Mitochondrial DNA analyses of Bronze-Age remains in northwestern China, Xinjiang (Zhao, 1998) and studies involving blood group antigens (Francalacci, 1998) support this hypothesis.
Even under the most adverse conditions, tissue derived from skeletal and dental remains generally contain fewer polymerase chain reaction (PCR) inhibitors than do soft tissue remains from the same specimen (Lassen et al., 1994). Comparative assays of both skeletal and soft tissue taken from Pre-Columbian South America mummies by Lassen et al., (1994), suggest that ancient DNA should preferentially be extracted from hard rather than soft tissues.
In dental remains, hydroxyapatite, the inorganic component of osseous tissue to which DNA preferentially binds, is present in higher concentrations than in skeletal remains. Furthermore, as teeth are considerably less susceptible to co-extracted contamination than skeletal remains (Zierdt, Hummel, Herrman, 1996), use of dental remains for ancient DNA analyses obviates one of ancient DNA study's most problematic concerns (Hagelberg, 1994).
BRIEF SUMMARY OF THE INVENTION
Since as little as 0.01 of a gram of dentin is required to yield aDNA of sufficient molecular height for study (DeGusta, Cook, Sensaubaugh, 1994), the Reverse-Root-Canal technique presented herein offers a non-destructive alternative to the methods currently practiced. Moreover, in addition to its ability to conserve much of the roots and the entire crown, the acquisition of aDNA by this method greatly curtails the amount of co-extracted contaminants (Smith et al., 1993).
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Merriwether et al. “Genetic variation in the New World: ancient teeth, bone, and tissue as sources of DNA” Birkhauser Verlag Basel Experientia, vol. 50, No. 6, p. 592-601, Jun. 1994.*
Woodward et al. “Amplification of Ancient Nuclear DNA from teeth and soft tissues” PCR Methods and Applications, vol. 3, p. 244-247, 1994.*
Mornstad et al. “Demonstration and semi-quantification of mtDNA from human dentine and its relation to age” Int. J. Legal Med. vol. 112, p. 98-100, 1999.*
Zierdt et al. “ Amplification of human STR from medieval teeth and bone samples” Human Biology, vol. 68, No. 2, p. 185-199, Apr. 1996.
Arthur Lisa B.
Goldberg Jeanine
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