Extraction Protocol for Parallel Analysis of Proteins and DNA from Ancient Teeth and Dental Calculus.

Dental calculus is becoming a crucial material in the study of past populations with increasing interest in its proteomic and genomic content. Here, we suggest further development of a protocol for analysis of ancient proteins and a combined approach for subsequent ancient DNA extraction. We tested the protocol on recent teeth, and the optimized protocol was applied to ancient tooth to limit the destruction of calculus as it is a precious and irreplaceable source of dietary, microbiological, and ecological information in the archeological context. Finally, the applicability of the protocol was demonstrated on samples of the ancient calculus.

The Application of Scanning Electron Microscopy with Energy-Dispersive X-Ray Spectroscopy (SEM-EDX) in Ancient Dental Calculus for the Reconstruction of Human Habits.

The great potential of scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM-EDX) is in detection of unusual chemical elements included in ancient human dental calculus to verify hypotheses about life and burial habits of historic populations and individuals. Elemental spectra were performed from archeological samples of three chosen individuals from different time periods. The unusual presence of magnesium, aluminum, and silicon in the first sample could confirm the hypothesis of high degree of dental abrasion caused by particles from grinding stones in flour. In the second sample, presence of copper could confirm that bronze jewelery could lie near the buried body. The elemental composition of the third sample with the presence of lead and copper confirms the origin of individual to Napoleonic Wars because the damage to his teeth could be explained by the systematic utilization of the teeth for the opening of paper cartridges (a charge with a dose of gunpowder and a bullet), which were used during the 18th and the 19th century AD. All these results contribute to the reconstruction of life (first and third individual) and burial (second individual) habits of historic populations and individuals.

Cost-effective straightforward method for captured whole mitogenome sequencing of ancient DNA.

Working with mitochondrial DNA from highly degraded samples is challenging. We present a whole mitogenome Illumina-based sequencing method suitable for highly degraded samples. The method makes use of double-stranded library preparation with hybridization-based target enrichment. The aim of the study was to implement a new user-friendly method for analysing many ancient DNA samples at low cost. The method combines the Swift 2S™ Turbo library preparation kit and xGen® panel for mitogenome enrichment. Swift allows to use low input of aDNA and own adapters and primers, handles inhibitors well, and has only two purification steps. xGen is straightforward to use and is able to leverage already pooled libraries. Given the ancient DNA is more challenging to work with, the protocol was developed with several improvements, especially multiplying DNA input in case of low concentration DNA extractions followed by AMPure® beads size selection and real-time pre-capture PCR monitoring in order to avoid cycle-optimization step. Nine out of eleven analysed samples successfully retrieved mitogenomes. Hence, our method provides an effective analysis of whole mtDNA, and has proven to be fast, cost-effective, straightforward, with utilisation in population-wide research of burial sites.

A very rare case of possible actinomycosis of the mandible from the Middle Ages.

OBJECTIVE: Documented cases of actinomycosis in archaeological skeletons are very rare, especially from Central Europe. Our contribution will help facilitate the differential diagnosis of this disease for other paleopathologists. MATERIAL: This paper describes a pathological finding of the skeleton of a 40-year-old male from a burial ground in Sady-Spitálky (Czech Republic) dated to the 10th-12th century. METHODS: The affected skeleton was evaluated as a probable case of actinomycosis on the basis of a detailed macroscopic, X-ray and histological examination. The osteolytic foci examined were compared with similar changes caused by tuberculosis, syphilis and mycoses. RESULTS: The character and location of the defect on the mandible is indicative of organ actinomycosis and is also reflected by the lytic lesion observed on a lumbar vertebra. CONCLUSIONS: The described case can be considered one of the very rare paleopathological findings of possible actinomycosis in humans in Central Europe. SIGNIFICANCE: Good evidence of bone actinomycosis findings may be beneficial for further paleopathological and epidemiological studies, especially for research focused on the diachronic development of actinomycosis in Europe. In doing so, all available factors, such as hygiene habits, nutrition, social structure and overall health of the population that could be causally related to its origin, course and treatment, can be taken into account. LIMITATIONS: The mandible of the studied individual was damaged, especially in the area affected by the lesion, so the paleopathological analysis was difficult to perform. SUGGESTION FOR THE FUTURE RESEARCH: In future, actinomycosis in this skeleton may be confirmed by bio-molecular analysis.

Influence of Magnetic Microparticles Isolation on Adenine Homonucleotides Structure.

The electroactivity of purine and pyrimidine bases is the most important property of nucleic acids that is very useful for determining oligonucleotides using square wave voltammetry. This study was focused on the electrochemical behavior of adenine-containing oligonucleotides before and after their isolation using paramagnetic particles. Two peaks were detected-peak A related to the reduction of adenine base and another peak B involved in the interactions between individual adenine strands and contributes to the formation of various spatial structures. The influence of the number of adenine bases in the strand in the isolation process using paramagnetic particles was investigated too.

Interaction of Heavy Metal Ions with Carbon and Iron Based Particles.

Due to the rapid development of industry and associated production of toxic waste, especially heavy metals, there is a great interest in creating and upgrading new sorption materials to remove these pollutants from the environment. This study aims to determine the effectiveness of different carbon forms (graphene, expanded carbon, multi-wall nanotubes) and paramagnetic particles (Fe2O3) for adsorption of cadmium(II), lead(II), and copper(II) on its surface, with different interaction time from 1 min to 24 h. The main attention is paid to the detection of these metals using differential pulse voltammetry. Based on the obtained results, graphene and Fe2O3 are found to be good candidates for removal of heavy metals from the environment.