Towards multi-phase postmortem CT angiography in children: a study on a porcine model.

PURPOSE: Multi-phase postmortem computed tomography angiography (MPMCTA) is a growing technique, which is standardized for adults. Application of this protocol for a children population is not so well defined. Our study aims to adapt the adult's protocol to children, using a porcine model. MATERIAL AND METHODS: Three groups of 18 pigs were studied, with a weight distribution between 4 and 48 kg. Different pump devices were used. Pigs of group I were studied using the Virtangio® machine, whereas pigs of groups II and III were studied using used the Medrad® machine. Study of vascular opacification was possible using a semi-quantitative method based on 26 arterial and 26 venous segments that were distributed over the entire body from the cephalic extremity to the posterior pawns. RESULTS: While thoracic, abdominal, and pelvic vascular opacification were complete for each individual pig in a group, group III showed better vascular opacification for the cephalic extremity. This was also true for anterior and posterior pawns vascular opacification. Spearman correlation tests showed a significant relationship between anthropometric characteristics of pigs, injection parameters, and percentage of opacified segments. A higher percentage of opacification was obtained for individuals of lower weights, with comparatively lower quantities of contrast agent injected. CONCLUSION: Postmortem computed tomography angiography (PMCTA) was possible for all the individuals, particularly for small weights (4 kg) using the Medrad® machine. However, further studies are needed to better understand the procedure.

Old hearts for modern investigations: CT and MR for archaeological human hearts remains.

INTRODUCTION: Among 800 burials dated between the 15th and 18th centuries and found in the center of Rennes (Brittany, France), a collection of five heart-shaped lead urns was discovered. This material was studied using classical methods (external study, autopsy and histology), and also modern imaging like computed tomography (CT), magnetic resonance (MR) before and after coronary opacification. The aim of this manuscript is to describe different steps of ancient soft tissues study, especially using imaging techniques. METHODS: The study gathered various specialists: anthropologists, archeologists, forensic pathologists, radiologists, pathologic physicians, and physicists. Imaging techniques were performed, before and after coronary opacification. Finally, hearts were autopsied and different histological samples were analyzed. RESULTS: Only heart n°2 was too damaged to be studied. Heart n°3 was considered as normal using all investigation techniques. The study of Hearts n°s 4 and 5 revealed dilated cardiomyopathy while Heart n°1 showed important signs of diffuse hypertrophic cardiomyopathy. Different fibro lipid plaques were identified using imaging techniques, and were confirmed by histology. CONCLUSIONS: The study of archeological soft tissues using modern imaging is possible if the material is well-preserved. This type of research can uncover principal findings, allowing scientists to establish diseases of ancient times.

Diagnostic value of high-resolution peripheral quantitative computed tomography (HR-pQCT) in the qualitative assessment of cribra orbitalia: a preliminary study.

Cribra orbitalia are a porotic or sieve-like lesions in the bony orbital roof. This characteristic has frequently been detected in palaeopathological skulls from many parts of the world and has been the object of extensive research. Our objective was to determine if high-resolution peripheral quantitative computed tomography (HR-pQCT) could produce reliable information in the study of cribra orbitalia. Seven skulls displaying cribra orbitalia were investigated by HR-pQCT. The two-dimensional slices were compared with histological sections. The HR-pQCT images and histological sections showed similar results, i.e. two groups of lesions with different characteristics. HR-pQCT can be of great value in palaeopathological research. It is a nondestructive, fast and precise technique that allows an easy evaluation of the bone architecture without destruction of the sample.

Novel contribution on the diagenetic physicochemical features of bone and teeth minerals, as substrates for ancient DNA typing.

The extraction of DNA from skeletal remains is a major step in archeological or forensic contexts. However, diagenesis of mineralized tissues often compromises this task although bones and teeth may represent preservation niches allowing DNA to persist over a wide timescale. This exceptional persistence is not only explained on the basis of complex organo-mineral interactions through DNA adsorption on apatite crystals composing the mineral part of bones and teeth but is also linked to environmental factors such as low temperatures and/or a dry environment. The preservation of the apatite phase itself, as an adsorption substrate, is another crucial factor susceptible to significantly impact the retrieval of DNA. With the view to bring physicochemical evidence of the preservation or alteration of diagenetic biominerals, we developed here an analytical approach on various skeletal specimens (ranging from ancient archeological samples to recent forensic specimens), allowing us to highlight several diagenetic indices so as to better apprehend the complexity of bone diagenesis. Based on complementary techniques (X-ray diffraction (XRD), Fourier transform infrared (FTIR), calcium and phosphate titrations, SEM-EDX, and gravimetry), we have identified specific indices that allow differentiating 11 biological samples, primarily according to the crystallinity and maturation state of the apatite phase. A good correlation was found between FTIR results from the analysis of the v3(PO4) and v4(PO4) vibrational domains and XRD-based crystallinity features. A maximal amount of information has been sought from this analytical approach, by way of optimized posttreatment of the data (spectral subtraction and enhancement of curve-fitting parameters). The good overall agreement found between all techniques leads to a rather complete picture of the diagenetic changes undergone by these 11 skeletal specimens. Although the heterogeneity and scarcity of the studied samples did not allow us to seek direct correlations with DNA persistence, the physicochemical parameters described in this work permit a fine differentiation of key properties of apatite crystals among post mortem samples. As a perspective, this analytical approach could be extended to more numerous sets of specimens so as to draw statistical relationships between mineral and molecular conservation.

Shotgun microbial profiling of fossil remains.

Millions to billions of DNA sequences can now be generated from ancient skeletal remains thanks to the massive throughput of next-generation sequencing platforms. Except in cases of exceptional endogenous DNA preservation, most of the sequences isolated from fossil material do not originate from the specimen of interest, but instead reflect environmental organisms that colonized the specimen after death. Here, we characterize the microbial diversity recovered from seven c. 200- to 13 000-year-old horse bones collected from northern Siberia. We use a robust, taxonomy-based assignment approach to identify the microorganisms present in ancient DNA extracts and quantify their relative abundance. Our results suggest that molecular preservation niches exist within ancient samples that can potentially be used to characterize the environments from which the remains are recovered. In addition, microbial community profiling of the seven specimens revealed site-specific environmental signatures. These microbial communities appear to comprise mainly organisms that colonized the fossils recently. Our approach significantly extends the amount of useful data that can be recovered from ancient specimens using a shotgun sequencing approach. In future, it may be possible to correlate, for example, the accumulation of postmortem DNA damage with the presence and/or abundance of particular microbes.

The rediscovery of smallpox.

Smallpox is an infectious disease that is unique to humans, caused by a poxvirus. It is one of the most lethal of diseases; the virus variant Variola major has a mortality rate of 30%. People surviving this disease have life-long consequences, but also assured immunity. Historically, smallpox was recognized early in human populations. This led to prevention attempts--variolation, quarantine, and the isolation of infected subjects--until Jenner's discovery of the first steps of vaccination in the 18th century. After vaccination campaigns throughout the 19th and 20th centuries, the WHO declared the eradication of smallpox in 1980. With the development of microscopy techniques, the structural characterization of the virus began in the early 20th century. In 1990, the genomes of different smallpox viruses were determined; viruses could be classified in order to investigate their origin, diffusion, and evolution. To study the evolution and possible re-emergence of this viral pathogen, however, researchers can only use viral genomes collected during the 20th century. Cases of smallpox in ancient periods are sometimes well documented, so palaeomicrobiology and, more precisely, the study of ancient smallpox viral strains could be an exceptional opportunity. The analysis of poxvirus fragmented genomes could give new insights into the genetic evolution of the poxvirus. Recently, small fragments of the poxvirus genome were detected. With the genetic information obtained, a new phylogeny of smallpox virus was described. The interest in conducting studies on ancient strains is discussed, in order to explore the natural history of this disease.

Virtual anthropology and forensic identification using multidetector CT.

Virtual anthropology is made possible by modern cross-sectional imaging. Multislice CT (MSCT) can be used for comparative bone and dental identification, reconstructive identification and lesion identification. Comparative identification, the comparison of ante- and post-mortem imaging data, can be performed on both teeth and bones. Reconstructive identification, a considerable challenge for the radiologist, identifies the deceased by determining sex, geographical origin, stature and age at death. Lesion identification combines virtual autopsy and virtual anthropology. MSCT can be useful in palaeopathology, seeking arthropathy, infection, oral pathology, trauma, tumours, haematological disorders, stress indicators or occupational stress in bones and teeth. We examine some of the possibilities offered by this new radiological subspeciality that adds a new dimension to the work of the forensic radiologist. A multidisciplinary approach is crucial and involves communication and data exchange between radiologists, forensic pathologists, anthropologists and radiographers.

Study of dental caries and periapical lesions in a mediaeval population of the southwest France: differences in visual and radiographic inspections.

Dental caries and periapical lesions have often been studied in archaeological samples. The majority of these studies concern lesions detected clinically and, in some cases, radiography is used to improve the scoring results. The purpose of the present study was to compare and combine the two methods for recording caries and cysts. The studied dental material derived from a mediaeval sample from the south of France. The study included sixty mandibles (788 teeth). Each tooth was observed clinically and radiographically. Absence of teeth was noted and unerupted teeth, which could be viewed on radiographs were also noted. The location and degree of development of each lesion were recorded. Occlusal lesions of degree 1 were mostly detected clinically (5.1% vs.1.4% detected radiographically). Radiographic detection increased the score of approximal lesions of degree 2 (1.4% vs. 0.7%) and the detection of intraosseous lesions. This research has shown that the combination of radiographic and clinical examinations improves the caries detection and thus, increases the caries score in a studied dental sample.

mtDNA variation in the Buryat population of the Barguzin Valley: New insights into the micro-evolutionary history of the Baikal area.

BACKGROUND: Southern Siberian populations, including the Buryat, have been of great interest in investigating the exchanges between Eastern and Western Eurasia and understanding the peopling of Siberia and the New World. AIM: Previous studies mainly employed a phylogenetic approach, and thus used pooled samples to detect a maximum of variability. As different sampling strategies may result in different pictures of a population's evolutionary history, we proposed in this study to focus on a local Buryat population selected on the basis of geographical, archaeological and ethno-historical data. SUBJECTS AND METHODS: This study investigated a local population from the Barguzin Valley, on the north-western shores of Lake Baikal identified as the most likely place of Buryat origin. We analysed mitochondrial DNA (mtDNA) RFLPs markers, HVS-I and HVS-II sequences to discuss the genetic variability of this population, and to compare our local sample with pooled Buryat samples and neighbouring Siberian populations. RESULTS: The Barguzin Buryat sample shows depressed neutrality scores compared to the pooled Buryat sample, and different genetic affinities with the Mongol and Turco-Evenk populations. CONCLUSION: These results underline the need to use local samples, in addition to pooled samples, to investigate the history of human populations at the micro-evolutionary level.

Dental caries, tooth wear and diet in an adult medieval (12th-14th century) population from mediterranean France.

OBJECTIVES: The aims of the present work were to determine the frequency and distribution of caries and tooth wear on paired maxillae of a mediaeval sample from southwest France in which the sex of the remains had been established, and to make a relation with the diet of this population. MATERIALS AND METHODS: The sample analysed consisted of the dental remains of 58 adult individuals (29 men and 29 women) excavated from the mediaeval cemetery of the archaeological site of Vilarnau d'Amont (southwest France). A total of 1395 teeth were examined. RESULTS: The frequency of ante-mortem tooth loss for the sample was 8.7% and the frequency of caries was 17.5%. The frequencies of carious lesions in adult men and women's dentition were 21.9% and 14.0%. The most frequent were occlusal (49.7% and 34.3%) and approximal caries (26.5% and 37.4%). Concerning tooth wear, all 58 individuals were affected by attrition (100%) and more than 90% of the teeth were concerned. Most of them showed the presence of dentin clusters. There was no significant difference between men and women for caries and tooth wear. DISCUSSION: These findings are similar to those of other studies on European populations of the same socio-economic status and confirm the predominance of tooth wear over carious lesions during this period. Both caries and tooth wear may be related to the regional diet of this rural population.

Uniparental (mtDNA, Y-chromosome) polymorphisms in French Guiana and two related populations--implications for the region's colonization.

Blood samples collected in four Amerindian French Guiana populations (Palikur, Emerillon, Wayampi and Kali'na) in the early 1980s were screened for selected mtDNA and Y-chromosome length polymorphisms, and sequenced for the mtDNA hypervariable segment I (HVS-I). In addition, two other Amerindian populations (Apalaí and Matsiguenga) were examined for the same markers to establish the genetic relationships in the area. Strong dissimilarities were observed in the distribution of the founding Amerindian haplogroups, and significant p-values were obtained from F(ST) genetic distances. Interpopulation similarities occurred mainly due to geography. The Palikur did not show obvious genetic similarity to the Matsiguenga, who speak the same language and live in a region from where they could have migrated to French Guiana. The African-origin admixture observed in the Kali'na probably derives from historical contacts they had with the Bushinengue (Noir Marron), a group of escaped slaves who now lead independent lives in a nearby region. This analysis has identified significant clues about the Amerindian peopling of the North-East Amazonian region.

First successful assay of Y-SNP typing by SNaPshot minisequencing on ancient DNA.

In the present study, a set of 13 Y-chromosomal single nucleotide polymorphisms (Y-SNPs) selected for the identification of the most frequent Asian Y-haplogroups was included in an allele-specific primer extension assay. Single nucleotide polymorphism (SNP) genotyping was accomplished by co-amplification of these 13 DNA fragments within 2 multiplex PCRs followed by detection with 1 minisequencing reaction using the SNaPshottrade mark Multiplex kit and analysis of extension products by capillary electrophoresis. First developed on modern samples, the assay was optimized for the analysis of 11 ancient DNA (aDNA) samples from the Krasnoyarsk region (southern Siberia) that were dated from 5,500-1,800 years before present (YBP). SNP typing was successful for most of them, which were all assigned to Y-haplogroup R1a1 except one. These results show that SNPs are well-suited for the analysis of aged and degraded DNA samples. Moreover, we found that the SNaPshot minisequencing methodology is a convenient, robust, and efficient method for SNP typing. To our knowledge, this study reports the first successful investigation of Y-SNPs on aDNA samples. The potential use of Y-SNPs in both evolutionary and forensic fields is also discussed.

STR typing of ancient DNA extracted from hair shafts of Siberian mummies.

The aim of this study was to determine if ancient hair shafts could be suitable for nuclear DNA analysis and to develop an efficient and straightforward protocol for DNA extraction and STR typing of ancient specimens. The developed method was validated on modern and forensic samples and then successfully applied on ancient hairs collected from Siberian mummies dating from the 16th to the early 19th centuries. In parallel extractions including or excluding a washing step were performed at least two times for each sample in order to evaluate the influence on the quantity of nuclear DNA yielded and on the typing efficiency. Twelve ancient individuals were analyzed through our approach and full and reliable profiles were obtained for four of them. These profiles were validated by comparison with those obtained from bone and teeth DNA extracted from the same ancient specimens. The present study demonstrates that the washing step cannot be considered as deleterious for DNA retrieval since the same results were obtained by the two approaches. This finding challenges the hypothesis that recoverable nuclear DNA is only found on the outer surface of hair shafts and provides evidence that nuclear DNA can be successfully extracted from ancient hair shafts. The method described here constitutes a promising way for non-invasive investigations in ancient DNA analysis for precious or historical samples as well as forensic casework analyses.

Early influence of the steppe tribes in the peopling of Siberia.

The Yakuts, Middle Age Turkic speakers (15th-16th centuries), are widely accepted as the first settlers of the Altai-Baikal area in eastern Siberia. They are supposed to have introduced horses and developed metallurgy in this geographic area during the 15th or 16th century A.D. The analysis of the Siberian grave of Pokrovsk, recently discovered near the Lena River (61 degrees 29' N) and dated by accelerator mass spectrometry from 2,400 to 2,200 years B.P., may provide new elements to test this hypothesis. The exceptional combination of various artifacts and the mitochondrial DNA data extracted from the bone remains of the Pokrovsk man might prove the existence of previous contacts between autochthonous hunters of Oriental Siberia and the nomadic horse breeders from the Altai-Baikal area (Mongolia and Buryatia). Indeed, the stone arrowhead and the harpoons relate this Pokrovsk man to the traditional hunters of the Taiga. Some artifacts made of horse bone and the pieces of armor, however, are related to the tribes of Mongolia and Buryatia of the Xiongnu period (3rd century B.C.). This affinity has been confirmed by the match of the mitochondrial haplotype of this subject with a woman of the Egyin Gol necropolis (Mongolia, 2nd/3rd century A.D.) as well as with two modern Buryats. This result allows us to postulate that contacts between southern steppe populations and Siberian tribes occurred before the 15th century.

Mitochondrial DNA analysis of horses recovered from a frozen tomb (Berel site, Kazakhstan, 3rd Century BC).

Sequence polymorphism of the mitochondrial DNA D-loop was used to determine the genetic diversity of horses recovered from a Scythian princely tomb dating from the beginning of the 3rd century BC. Eight haplotypes were found among the 13 ancient horse samples tested. Phylogenetical analysis showed that these ancient horse's sequences, along with two Yakut ones, were distributed throughout the tree defined by modern horses' sequences and are closely related to them. No clear geographical affiliation of the specimens studied was thus determined. Our work, among others, supports the very ancient origin of the matrilines in horses.

[Shamanic healing in the age of modern medicine]. / Medecine chamanique et modernite medicale.

This study on the therapeutic aspects of shamanic medicine differentiates the anthropologic approaches used in the 19th and early 20th century from contemporary approaches developed since the n Central to Eastern Asia. In addition to reviewing the major works on shamanic practice, this study takes into account lesser-known data on contemporary shamanic practices in Nepal. Shamanism embraces special cosmogenic beliefs. From a biological standpoint the shamanic trance like various healing techniques raises interesting questions about the involvement of certain brain regions in the development of mental factors that may generate somatic symptoms. From the therapeutic standpoint it shows the importance of managing social and psychoaffective factors. The current resurgence of shamanism could signal the revival of the disrupted ethnic identities. The shaman and medical doctor occupy symmetric, inversed positions. The shaman is a master in the management of social and psychoaffective health determinants. Until now the physician may have tended to overlook the human side of disease.

Genetic analysis of a Scytho-Siberian skeleton and its implications for ancient Central Asian migrations.

The excavation of a frozen grave on the Kizil site (dated to be 2500 years old) in the Altai Republic (Central Asia) revealed a skeleton belonging to the Scytho-Siberian population. DNA was extracted from a bone sample and analyzed by autosomal STRs (short tandem repeats) and by sequencing the hypervariable region I (HV1) of the mitochondrial DNA. The resulting STR profile, mitochondrial haplotype, and haplogroup were compared with data from modern Eurasian and northern native American populations and were found only in European populations historically influenced by ancient nomadic tribes of Central Asia.

Genetic analysis of human remains from a double inhumation in a frozen kurgan in Kazakhstan (Berel site, Early 3rd Century BC).

The discovery of a big barrow of the Saka period in eastern Kazakhstan between the Russian and the Chinese borders provided the opportunity to excavate a frozen burial site. In the burial chamber, there was a wooden sarcophagus with two human bodies. The skeletons of these two individuals, a man and a woman, were well preserved. A genetic study based on STRs and mitochondrial DNA analyses was undertaken in order to determine whether these human remains belonged to close relatives. Results were obtained for all the markers. Nevertheless, nuclear STRs did not allow a clear conclusion concerning the relationship, but analysis of mitochondrial DNA showed that these skeletons were not close relatives.

Molecular identification by "suicide PCR" of Yersinia pestis as the agent of medieval black death.

Medieval Black Death is believed to have killed up to one-third of the Western European population during the 14th century. It was identified as plague at this time, but recently the causative organism was debated because no definitive evidence has been obtained to confirm the role of Yersinia pestis as the agent of plague. We obtained the teeth of a child and two adults from a 14th century grave in France, disrupted them to obtain the pulp, and applied the new "suicide PCR" protocol in which the primers are used only once. There were no positive controls: Neither Yersinia nor Yersinia DNA were introduced in the laboratory. A negative result is followed by a new test using other primers; a positive result is followed by sequencing. The second and third primer pair used, coding for a part of the pla gene, generated amplicons whose sequence confirmed that it was Y. pestis in 1 tooth from the child and 19/19 teeth from the adults. Negative controls were negative. Attempts to detect the putative alternative etiologic agents Bacillus anthracis and Rickettsia prowazekii failed. Suicide PCR avoids any risk of contamination as it uses a single-shot primer-its specificity is absolute. We believe that we can end the controversy: Medieval Black Death was plague.

[Stomatologic and maxillofacial pathology in a medieval population (10th-12th centuries) of southwestern France]. / Etude de la pathologie stomatologique et maxillo-faciale dans une population médiévale (Xe-XIIe siècles) du sud-ouest de la France.

We present an assessment of the dental and maxillofacial pathology in a medieval population in southwestern France. One hundred and ninety eight mandibles and 29 craniofacial complexes were analysed. Dental and periodontal infectious pathology predominated. Third molar agenesia was quite frequent, concerning 25% of the mandibles. Third molar eruption was almost constant and in a normal position. Condylar process degeneration concerned 6% of the population. Three cases of traumatic pathology were observed, one case of long mandible was noted, and two cases of hypertrophic inferior alveolar process. Dento-mandibular maladjustment was uncommon. No unwedging of the maxillo-mandibular bone basis was observed.