RESUMO
BACKGROUND: Recent advances in the next-generation sequencing (NGS) allowed the metagenomic analyses of DNA from many different environments and sources, including thousands of years old skeletal remains. It has been shown that most of the DNA extracted from ancient samples is microbial. There are several reports demonstrating that the considerable fraction of extracted DNA belonged to the bacteria accompanying the studied individuals before their death. RESULTS: In this study we scanned 344 microbiomes from 1000- and 2000- year-old human teeth. The datasets originated from our previous studies on human ancient DNA (aDNA) and on microbial DNA accompanying human remains. We previously noticed that in many samples infection-related species have been identified, among them Tannerella forsythia, one of the most prevalent oral human pathogens. Samples containing sufficient amount of T. forsythia aDNA for a complete genome assembly were selected for thorough analyses. We confirmed that the T. forsythia-containing samples have higher amounts of the periodontitis-associated species than the control samples. Despites, other pathogens-derived aDNA was found in the tested samples it was too fragmented and damaged to allow any reasonable reconstruction of these bacteria genomes. The anthropological examination of ancient skulls from which the T. forsythia-containing samples were obtained revealed the pathogenic alveolar bone loss in tooth areas characteristic for advanced periodontitis. Finally, we analyzed the genetic material of ancient T. forsythia strains. As a result, we assembled four ancient T. forsythia genomes - one 2000- and three 1000- year-old. Their comparison with contemporary T. forsythia genomes revealed a lower genetic diversity within the four ancient strains than within contemporary strains. We also investigated the genes of T. forsythia virulence factors and found that several of them (KLIKK protease and bspA genes) differ significantly between ancient and modern bacteria. CONCLUSIONS: In summary, we showed that NGS screening of the ancient human microbiome is a valid approach for the identification of disease-associated microbes. Following this protocol, we provided a new set of information on the emergence, evolution and virulence factors of T. forsythia, the member of the oral dysbiotic microbiome.
Assuntos
Restos Mortais/microbiologia , Fósseis/microbiologia , Microbioma Gastrointestinal , Boca/microbiologia , Tannerella forsythia/genética , Tannerella forsythia/patogenicidade , Fatores de Virulência/genética , Genoma Bacteriano , Genômica , Humanos , Metagenoma , Periodontite/microbiologia , Periodonto/microbiologia , Dente/microbiologiaRESUMO
The paper presents a comparative analysis of the circular and flat cross-section belts using measurements of a set of thermomechanical parameters, contributing to research about hot plate welding of drive belts. On the basis of thermogravimetric and spectrophotometric tests, information about the same chemical composition of the two belts was obtained. Dynamic thermomechanical analysis and scanning differential calorimetry provided information about a small difference between belts, which disappeared when the material was placed in a state of increased temperature and mechanical stress. On the basis of the analysis of the specific heat, thermal diffusion, density, and hardness, the values of the selected thermal properties of the belt were obtained, and a large similarity between the belts was identified. On the basis of the novel performed test cycle, it has been hypothesized that circular and flat belts made from thermoplastic polyurethane elastomer could be used interchangeably for butt-welding testing. It has also been proven that cyclic thermomechanical loads unify the properties of both materials so that multiple mechanical and thermal loads do not result in any change in the material properties of the two belts. As a consequence, changes in the weld properties after welding, compared to a solid belt, are not expected.
RESUMO
deltaF508 is the most common (70%) among over 1000 mutations of the gene encoding ATP-regulated chloride channel, namely CFTR--cystic fibrosis transmembrane regulator. The time which passed from the calculated mutation event was anticipated on the basis of the frequency of contemporary haplotypes, but not on its direct identification. The presence of three base pairs deletion in the ancient DNA (aDNA) isolated from skeletal remains of the Middle Ages origin was investigated. Teeth excavated in the area of three sites located in Central Poland were processed for a DNA. 6 out of 82 samples did not produce amplificable fragments of DNA. Although the number of specimens analyzed was sufficient to confirm the presence of the rare mutation, the deltaF508 CFTR sequence was not found in the remains of individuals living back 35 - 45 generations. The absence of the mutated allele in the particular geographic region cannot state for the status of mutated allele throughout the country, especially at times when migrations were limited and movements of people were more area restricted than at present days.