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DystrophicThus, the EB panel sequencing of genomic DNA, followed by a paternity test and analysis of microsatellite markers, as well as multiplex ligation-dependent probe amplification (MLPA) copy number analysis using patient and parental DNA, were performed.This approach revealed a paternally derived hemizygous deletion spanning from exon 3 to exon 118. Linear amplification-mediated PCR (LAM-PCR) determined the breaking points within intron 2 of the COL7A1 gene, comprising a 40kb segment within intron 1 of the adjacentThis report highlights the relevance of advanced molecular profiling to determine new/exceptional/unusual genotypes and the accurate mode of genetic transmission in DEB.

The Deputy Führer of the Third Reich Rudolf Hess was captured after a controversial flight to Scotland in 1941. Hess was sentenced to life imprisonment during the Nuremberg War Crimes Trials. He was detained in Berlin's Spandau Prison under the official security designation 'Spandau #7.' Early doubts arose about the true identity of prisoner 'Spandau #7.' This evolved to a frequently espoused conspiracy theory that prisoner 'Spandau #7' was an imposter and not Rudolf Hess. After Hess's reputed 1987 suicide, the family grave became a Neo-Nazi pilgrimage site. In 2011, the grave was abandoned and the family remains cremated. Here we report the forensic DNA analysis of the only known extant DNA sample from prisoner 'Spandau #7' and a match to the Hess male line, thereby refuting the Doppelgänger Theory.

Archaeological excavations at Punta di Zambrone (Calabria, Italy) lead to the recovery of severely fragmented human remains in a fortification ditch (Area C). Besides other findings like animal bones and artefacts, the filling of this ditch also contained a large amount of plant ash, indicative of a prolonged period of fire-related processes, dating back to the 2nd millennium BC. Comprehensive anthropological investigations were carried out to determine the sex and age as well as the health status of the discarded individuals. Apart from this, a set of methods from forensic anthropology were applied to reconstruct the circumstances of death and related taphonomic processes. The reassociation of skeletal fragments from the ditch revealed a minimum number of two individuals. Most of the bone fragments were assigned to an early adult individual of questionable sex. The remaining fragments could be tentatively assigned to a slightly younger (adolescent) individual. However, for the adolescent individual, the low number of associated bone fragments rendered macroscopic sexing elusive. Dental enamel hypoplasias were present on one premolar, indicating stress-induced perturbation of crown formation during childhood for one of the individuals. In addition, the same individual also displayed new bone formation around the external auditory pore (porus acusticus externus) in the tympanic portion of the temporal bone, suggestive of an inflammatory process affecting the external ear. Remarkably, none of the human bone fragments exhibited burn marks. This finding rules out the possibility of a direct link with the assumed burning event. Taphonomic reconstruction of the excavation site suggests that the remains of at least one of the excavated individuals have undergone multiple disturbances after deposition.

Summary We subjected human paleofeces dating from the Bronze Age to the Baroque period (18th century AD) to in-depth microscopic, metagenomic, and proteomic analyses. The paleofeces were preserved in the underground salt mines of the UNESCO World Heritage site of Hallstatt in Austria. This allowed us to reconstruct the diet of the former population and gain insights into their ancient gut microbiome composition. Our dietary survey identified bran and glumes of different cereals as some of the most prevalent plant fragments. This highly fibrous, carbohydrate-rich diet was supplemented with proteins from broad beans and occasionally with fruits, nuts, or animal food products. Due to these traditional dietary habits, all ancient miners up to the Baroque period have gut microbiome structures akin to modern non-Westernized individuals whose diets are also mainly composed of unprocessed foods and fresh fruits and vegetables. This may indicate a shift in the gut community composition of modern Westernized populations due to quite recent dietary and lifestyle changes. When we extended our microbial survey to fungi present in the paleofeces, in one of the Iron Age samples, we observed a high abundance of Penicillium roqueforti and Saccharomyces cerevisiae DNA. Genome-wide analysis indicates that both fungi were involved in food fermentation and provides the first molecular evidence for blue cheese and beer consumption in Iron Age Europe., Graphical abstract, Highlights • Gut microbiome and diet of European salt miners determined using paleofeces • Until the Baroque, the microbiome resembled that of modern non-Westernized people • Food-fermenting fungi in Iron Age feces indicates blue cheese and beer consumption, Maixner et al. describe the gut microbiome and diet of European salt miners using paleofeces dating from the Bronze Age to the Baroque period. This analysis provides evidence for recent changes in the gut microbiome due to industrialization and for the consumption of fermented food and beverages in Iron Age Europe.

When bodies are highly decomposed and identification based on morphologic features is no longer possible, short tandem repeat (STR) genotyping has been proved as a convenient and reliable alternative. However, at very advanced decomposition stages postmortal tissue putrefaction processes can affect microsatellite instability and therefore not all types of post mortem tissues are still equally suitable for STR genotyping and subsequent forensic identification. In many cases bone or dental material is used but here reprocessing is very elaborate and expensive. Thus, multiple studies have focused on the eligibility of soft tissue and organs where DNA extraction is a lot easier and faster. In this study different cases were evaluated in which a successful forensic identification was only possible with DNA material extracted from specific tissues. By doing so we aim to compare differential suitability of soft tissues for DNA analysis of highly decomposed bodies. This was achieved by a systematic classification of decomposed bodies and the analysis of multiple organs to evaluate the qualification of soft tissue for DNA profiling in respect to the degree of decomposition. In summary, we detected differences in the suitability of various soft tissues for DNA-based identification. Cerebral matter turned out to be one of the most useful targets in highly decomposed bodies.

In forensic genotyping the gender specific marker Amelogenin is a valuable tool for gender determination. This marker is therefore standard in all commercial forensic genetic kits. However, gonosomale aberrations, deletions or primer binding site mutations can cause problems in the correct interpretation of the marker Amelogenin and consequently a valid sex determination. Misleading results can be obtained based on incorrect interpretation of the phenotype or genetic intersexuality even though the genotyping based on Amelogenin itself was performed and evaluated correctly. In our routine casework we observed six cases with discrepancies regarding the biological and the “legal” gender. Further investigation of these cases revealed varying reasons for these discrepancies. But not all cases could be completely resolved and clarified.

With the new 6-dye AmpFISTR(®) Yfiler(®) Plus amplification kit (Thermo Fisher Scientific, Waltham, MA, USA) a set of 25 Y-chromosomal short tandem repeat loci (Y-STRs), including seven rapidly mutating Y-STRs (RM Y-STRs), is now available for forensic DNA typing. In this study we present our validation data for the AmpFISTR(®) Yfiler(®) Plus amplification kit and show the results of Y-chromosomal typing of 425 unrelated male individuals from two Austrian populations (Salzburg and Upper Austria) with the AmpFISTR(®) Yfiler(®) Plus amplification kit. Forensic parameters were calculated and compared for four Y-STR marker sets. We also typed five brother pairs to evaluate the power of discrimination for related individuals. The AmpFISTR(®) Yfiler(®) Plus (Yfiler Plus) kit appeared to be unimpaired by typical inhibitors such as hematin and humic acid or by large amounts of female components. An upgrade of analyzed markers resulted in increased discrimination capacity that is crucial for forensic trace analysis.

Forensic DNA analyses have become more and more sensitive in the past years. With the ability to generate DNA profiles even from minute amounts of cellular material also the possibility to detect DNA on trace material that originates from persons not linked to the crime event, such as crime scene investigators, increases. The contamination of crime scene samples can lead to false positive results and misinterpretation that can cause deceptive investigations. In this work we continue a study of 2010 that compared the number of detected contamination incidents that were caused in the pre-analytical phase of forensic DNA analysis with the number of crime scene samples analyzed by our laboratory. Within the past 17 years we were able to detect a total of 347 contamination incidents caused by police officers in approximately 46,000 trace samples to their origin (0.75%). Additionally we demonstrate the usefulness of reference profile databases that contain DNA profiles of police officers to detect contamination incidents of trace material.

Aim A collaborative exercise with several institutes was organized by the Forensic DNA Service (FDNAS) and the Institute of the Legal Medicine, 2nd Faculty of Medicine, Charles University in Prague, Czech Republic, with the aim to test performance of different laboratories carrying out DNA analysis of relatively old bone samples. Methods Eighteen laboratories participating in the collaborative exercise were asked to perform DNA typing of two samples of bone powder. Two bone samples provided by the National Museum and the Institute of Archaelogy in Prague, Czech Republic, came from archeological excavations and were estimated to be approximately 150 and 400 years old. The methods of genetic characterization including autosomal, gonosomal, and mitochondrial markers was selected solely at the discretion of the participating laboratory. Results Although the participating laboratories used different extraction and amplification strategies, concordant results were obtained from the relatively intact 150 years old bone sample. Typing was more problematic with the analysis of the 400 years old bone sample due to poorer quality. Conclusion The laboratories performing identification DNA analysis of bone and teeth samples should regularly test their ability to correctly perform DNA-based identification on bone samples containing degraded DNA and potential inhibitors and demonstrate that risk of contamination is minimized.

A badly decomposed body required identification by means of DNA analysis. A brother and sister of the deceased were available as reference subjects. Although investigation of Y-chromosomal markers established an exclusion condition, autosomal markers suggested a positive identification. In order to increase the reliability of the tests, X-chromosomal markers were also investigated. This analysis showed the body to have an XXY genotype (Klinefelter's syndrome). A number of hypotheses were assessed using biostatistical methods, ultimately resulting in a definite identification. The special aspect of Klinefelter's syndrome proved highly useful for biostatistical analysis.

Genetic information discovered, characterized for and used in forensic case-works and anthropology has shown to be also highly useful and relevant in investigating human remains from archaeological findings. By technical means, forensic and aDNA (ancient Deoxyribonucleic acid) analyses are well suited to be done using the same laboratory infrastructures and scientific expertise referring to sampling, sample protection, sample processing, contamination control as well as requiring analogous technical know how and knowledge on reading and interpreting DNA encoded information. Forensic genetics has significantly profited from aDNA-related developments (and vice versa, of course!), especially, when it comes to the identification of unknown human remains referring to the detection limit. Additionally the tremendous developments of analyzing chemistry and kits as well as instruments in forensics opened the whole panel of reading human and nonhuman DNA for historians and archaeologists but also for anthropologists. Ancient DNA / molecular archaeology, however, is not limited to the comparatively restrictive set of information as usually employed in forensic case work analyses but can also be applied to phenotypical markers, ethno-related genotypes or pathological features.In this review the authors give a general overview on the field of ancient DNA analysis focussing of the potentials and limits, fields of application, requirements for samples, laboratory setup, reaction design and equipment as well as a brief outlook on current developments, future perspectives and potential cross links with associated scientific disciplines.Key words: Human DNA, Ancient DNA, Forensic DNA typing, Molecular archaeology, Application.

Ancient DNA analyses have shown to be a powerful tool in the joint transdisciplinary assessment of archaeological records involving human remains. In this study we set out to identify single inhumations by synoptically evaluating the historical, archaeological, anthropological and molecular records on human remains from the crypt of the aristocratic family of Hoheneck (or: Hohenegg) dating to the 18(th) and 19(th) century AD. A total of 11 individuals were under investigation, yielding complete autosomal and Y-chromosomal STR profiles for 5 persons clearly showing a family group. DNA results, anthropological data and archaeological records taken together resulted in (almost) unambiguous correlation to historical records on the persons entombed in the crypt.

With the development of more sensitive DNA amplification and STR typing kits, the chance of detecting minute contamination of trace material increases. At the same time, possible ways of detecting unwanted contamination of crime scene samples were also enhanced. To estimate the contamination rate of biological traces nationwide, we continued a study from 2010 that compares the number of analyzed crime scene samples with the number of detected contamination incidents within our laboratory. The results of this study illustrate the importance of reference profiles, especially of crime scene investigators, and the advantage of database assisted profile alignment in cases of mixtures.

Due to its high reliability, DNA-typing is the method of preference in the field of osseous human remains identification. Nevertheless, contaminations from various sources have been shown to be inherent to the system, especially if the DNA-yield of samples under investigation is expected to be at a low level. For this reason a special focus has to be put on sampling procedures and contamination control in order to prevent from false results. In this study we present an illustrative case report followed by particular recommendations for taking samples from osseous human remains.

Y-STR markers are a valuable tool in the analysis of biological traces in which a mixture of male and female trace material is to be expected. It is possible to generate a Y-chromosome DNA profile, even if all the prior sperm tests are negative and no sign of any male component is found in amelogenin. In 38 of a total of 239 sexual offences a perpetrator trace was identified solely using Y-STR analysis. Based on these findings, the Austrian National DNA Database was expanded to include Y-STRs in 2012 with the primary objective to identify serial sexual offences.

The introduction of five "new European standard loci" (D1S1656, D2S441, D10S1248, D12S391 and D22S1045) in forensic DNA analysis called for the development of new Multiplex PCR kits. One of the aims was to combine the widespread SGM+ marker combination with the new markers to be amplified in a single kit. In addition, SE33 was also to be included in the new kits. We present practical experiences that have been made since the introduction of PowerPlex ESI 17 and AmpflSTR NGM SElect in our laboratory.