Pathogenic microbial ancient DNA: a problem or an opportunity?

Copyright © Royal Society 2006Eske Willerslev, Alan Coope

Confirmation of the presence of Mycobacterium-tuberculosis complex-specific DNA in three archaeological specimens

This journal published the first reported identification of Mycobacterium tuberculosis complex (MTE) DNA in ancient human remains but CONCERNS were raised about the article two years after publication. These were based on methodology which, in the field of ancient DNA, was still developing. Here we present a re-examination of the 1993 research conducted on three specimens which exhibited palaeopathologies indicative of tuberculosis. The specimens were: an ulna from pre-European-contact Borneo, a spine from Byzantine Turkey, and a lumbar-sacral spine from 17th century Scotland. There was insufficient material to permit re-examination of all of the original samples. The earlier results were confirmed in two independent laboratories using different methodologies. MTB DNA complex-specific Dna amplicons were obtained, and sequenced in both laboratories, in a re-analysis of samples which supported the earlier findings

Widespread occurrence of Mycobacterium tuberculosis DNA from 18th-19th century Hungarians

A large number (265) of burials from 1731-1838 were discovered in sealed crypts of the Dominican Church, Vac, Hungary in 1994. Many bodies were naturally mummified, so that both soft tissues and bones were available. Contemporary archives enabled the determination of age at death, and the identification of family groups. In some cases, symptoms before death were described and, occasionally, occupation. Initial radiological examination of a small number of individuals had indicated calcified lung lesions and demonstrable acid-fast bacteria suggestive of tuberculosis infection. Tuberculosis was endemic in 18th-19th century Europe, so human remains should contain detectable Mycobacterium tuberculosis complex (MTB) DNA, enabling comparisons with modern isolates. Therefore, a comprehensive examination of 168 individuals for the presence of MTB DNA was undertaken. Specific DNA amplification methods for MTB showed that 55% of individuals were positive and that the incidence varied according to age at death and sampling site in the body. Radiographs were obtained from 27 individuals and revealed an association between gross pathology and the presence of MTB DNA. There was an inverse relationship between PCR positivity and MTB target sequence size. In some cases, the preservation of MTB DNA was excellent, and several target gene sequences could be detected from the same sample. This information, combined with MTB DNA sequencing data and molecular typing techniques, will enable us to study the past epidemiology of TB infection, and extends the timeframe for studying changes in molecular fingerprints. Am J Phys Anthropol 120:144-152, 2003. (C) 2003 Wiley-Liss, Inc

Molecular analysis of Mycobacterium tuberculosis DNA from a family of 18th century Hungarians

The naturally mummified remains of a mother and two daughters found in an 18th century Hungarian crypt were analysed, using multiple molecular genetic techniques to examine the epidemiology and evolution of tuberculosis. DNA was amplified from a number of targets on the Mycobacterium tuberculosis genome, including DNA from IS6110, gyrA, katG codon 463, oxyR, dnaA–dnaN, mtp40, plcD and the direct repeat (DR) region. The strains present in the mummified remains were identified as M. tuberculosis and not Mycobacterium bovis, from katG and gyrA genotyping, PCR from the oxyR and mtp40 loci, and spoligotyping. Spoligotyping divided the samples into two strain types, and screening for a deletion in the MT1801–plcD region initially divided the strains into three types. Further investigation showed, however, that an apparent deletion was due to poor DNA preservation. By comparing the effect of PCR target size on the yield of amplicon, a clear difference was shown between 18th century and modern M. tuberculosis DNA. A two-centre system was used to confirm the findings of this study, which clearly demonstrate the value of using molecular genetic techniques to study historical cases of tuberculosis and the care required in drawing conclusions. The genotyping and spoligotyping results are consistent with the most recent theory of the evolution and spread of the modern tuberculosis epidemic

On Location: Theory and Practice in Classroom-Based Writing Tutoring

Classroom-based writing tutoring is a distinct form of writing support, a hybrid instructional method that engages multiple voices and texts within the college classroom. Tutors work on location in the thick of writing instruction and writing activity. On Location is the first volume to discuss this emerging practice in a methodical way. The essays in this collection integrate theory and practice to highlight the alliances and connections on-location tutoring offers while suggesting strategies for resolving its conflicts. Contributors examine classroom-based tutoring programs located in composition courses as well as in writing intensive courses across the disciplines. While earlier scholarship has focused on logistical and administrative matters, emphasizing, the worthiness of such programs and how to set them up, this volume asks further questions--questions that challenge and even critique classroom-based writing tutoring practices and principles. It poses new theories and offers alternative vantage points from which to reconsider long-standing theoretical controversies. At the same time, the contributors here are cognizant of newcomers\u27 questions regarding logistical/administrative issues, especially as configurations of classroom-based writing tutoring multiply. And in a concluding chapter, the editors suggest strategies for successfully implementing this important instructional practice, and propose future sites of theoretical and practical inquiry.https://digitalcommons.usu.edu/usupress_pubs/1150/thumbnail.jp

Long-Lasting Alterations in Membrane Properties, K+ Currents, and Glutamatergic Synaptic Currents of Nucleus Accumbens Medium Spiny Neurons in a Rat Model of Alcohol Dependence

Chronic alcohol exposure causes marked changes in reinforcement mechanisms and motivational state that are thought to contribute to the development of cravings and relapse during protracted withdrawal. The nucleus accumbens (NAcc) is a key structure of the mesolimbic dopaminergic reward system. Although the NAcc plays an important role in mediating alcohol-seeking behaviors, little is known about the molecular mechanisms underlying alcohol-induced neuroadaptive changes in NAcc function. The aim of this study was to investigate the effects of chronic intermittent ethanol (CIE) treatment, a rat model of alcohol withdrawal and dependence, on intrinsic electrical membrane properties and glutamatergic synaptic transmission of medium spiny neurons (MSNs) in the NAcc core during protracted withdrawal. We show that CIE treatment followed by prolonged withdrawal increased the inward rectification of MSNs observed at hyperpolarized potentials. In addition, MSNs from CIE-treated animals displayed a lower input resistance, faster action potentials (APs), and larger fast afterhyperpolarizations (fAHPs) than MSNs from vehicle-treated animals, all suggestive of increases in K+-channel conductances. Significant increases in the Cs+-sensitive inwardly rectifying K+-current accounted for the increased input resistance, while increases in the A-type K+-current accounted for the faster APs and increased fAHPs in MSNs from CIE rats. We also show that the amplitude and the conductance of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR)-mediated mEPSCs were enhanced in CIE-treated animals due to an increase in a small fraction of functional postsynaptic GluA2-lacking AMPARs. These long-lasting modifications of excitability and excitatory synaptic receptor function of MSNs in the NAcc core could play a critical role in the neuroadaptive changes underlying alcohol withdrawal and dependence