Biomolecular analyses enable new insights into ancient Egyptian embalming.

The ability of the ancient Egyptians to preserve the human body through embalming has not only fascinated people since antiquity, but also has always raised the question of how this outstanding chemical and ritual process was practically achieved. Here we integrate archaeological, philological and organic residue analyses, shedding new light on the practice and economy of embalming in ancient Egypt. We analysed the organic contents of 31 ceramic vessels recovered from a 26th Dynasty embalming workshop at Saqqara1,2. These vessels were labelled according to their content and/or use, enabling us to correlate organic substances with their Egyptian names and specific embalming practices. We identified specific mixtures of fragrant or antiseptic oils, tars and resins that were used to embalm the head and treat the wrappings using gas chromatography-mass spectrometry analyses. Our study of the Saqqara workshop extends interpretations from a micro-level analysis highlighting the socio-economic status of a tomb owner3-7 to macro-level interpretations of the society. The identification of non-local organic substances enables the reconstruction of trade networks that provided ancient Egyptian embalmers with the substances required for mummification. This extensive demand for foreign products promoted trade both within the Mediterranean8-10 (for example, Pistacia and conifer by-products) and with tropical forest regions (for example, dammar and elemi). Additionally, we show that at Saqqara, antiu and sefet-well known from ancient texts and usually translated as 'myrrh' or 'incense'11-13 and 'a sacred oil'13,14-refer to a coniferous oils-or-tars-based mixture and an unguent with plant additives, respectively.

Bacterial species-specific antimicrobial efficacies of three different body embalming solutions for anatomical studies.

While body decompensation is mainly facilitated by bacteria, investigating the antimicrobial properties of body preservation methods is still a neglected research area. We performed microbiological sampling for potentially pathogenic bacteria species of brain, lung, liver, colon, and subcutis samples obtained from bodies perfused with embalming solutions of variable composition with emphasis on variable formaldehyde concentrations. We, thereby, identified spore-forming aerobic and anaerobic bacteria mainly in the samples obtained from the colon of ethanol- and lower-concentrated formaldehyde formulation embalmed bodies. Moreover, we could identify Enterococcus species in bodies preserved with the latter method. Tissue samples of the subcutis remained sterile. Long-term incubation of special mycobacteria growth indicator tubes revealed no growth of mycobacteria in all 60 samples analyzed. Overall, we show survival of bacterial genera known to be especially environmentally resistant but also include potentially pathogenic members. Knowledge of bactericidal capacities of embalming solutions are therefore critical to assess risk and apply appropriate disinfection routines while working with human bodies. Moreover, new formulations to reduce potentially toxic substances for embalming needs to be evaluated regarding their bactericidal capacities.

A technique for decreasing reflection during cadaveric photography.

To create anatomical educational materials that can be viewed in three dimensions using stereo photographs and photogrammetry, multiple photographs must be taken from different directions. In this process, shadows and reflections from different positions in each photograph are undesirable for creating three-dimensional (3D) anatomy educational materials. Although a ring flash eliminates shadows, allowing light to enter from all directions, reflections cannot be eliminated. In particular, Thiel-embalmed cadavers, which are widely used in clinical anatomy, are highly wet and exhibit strong specular highlights. In this study, a straight polarization filter was attached to a handheld camera lens and ring flash, and shooting was performed using cross-polarization photography. Consequently, even in Thiel-embalmed cadavers, the details lost due to the effects of reflections and shadows can be recovered, and good results can be obtained when taking stereo photos or creating a 3D model using photogrammetry.

Development of the McMaster Embalming Scale (MES) to assess embalming solutions for surgical skills training.

Human cadavers used for surgical training are embalmed using various methods to facilitate tissue storage and longevity while preserving the natural characteristics required to achieve high fidelity functional task alignment. However, there are no standardized means to evaluate the suitability of embalming solutions for this purpose. The McMaster Embalming Scale (MES) was developed to assess the extent to which embalming solutions allow tissues to achieve physical and functional correspondence to clinical contexts. The MES follows a five-point Likert scale format and evaluates the effect of embalming solutions on tissue utility in seven domains. This study aims to determine the reliability and validity of the MES by presenting it to users after performing surgical skills on tissues embalmed using various solutions. A pilot study of the MES was conducted using porcine material. Surgical residents of all levels and faculty were recruited via the Surgical Foundations program at McMaster University. Porcine tissue was unembalmed (fresh- frozen) or embalmed using one of seven solutions identified in the literature. Participants were blinded to the embalming method as they completed four surgical skills on the tissue. After each performance, participants evaluated their experience using the MES. Internal consistency was evaluated using Cronbach's alpha. Domain to total correlations and a g-study were also conducted. Formalin-fixed tissue achieved the lowest average scores, while fresh frozen tissue achieved the highest. Tissues preserved using Surgical Reality Fluid (Trinity Fluids, LLC, Harsens Island, MI) achieved the highest scores among embalmed tissues. The Cronbach's alpha scores varied between 0.85 and 0.92, indicating a random sample of new raters would offer similar ratings using the MES. All domains except odor were positively correlated. The g-study indicated that the MES is able to differentiate between embalming solutions, but an individual rater's preference for certain tissue qualities also contributes to the variance in scores captured. This study evaluated the psychometric characteristics of the MES. Future steps to this investigation include validating the MES on human cadavers.

Deep Paleoproteotyping and Microtomography Revealed No Heart Defect nor Traces of Embalming in the Cardiac Relics of Blessed Pauline Jaricot.

Scientific examination of the heart of Blessed Pauline Jaricot-a French missionary figure-was carried out in 2022. As tandem mass spectrometry proteotyping has proven to be valuable to obtain the broad taxonomic repertoire of a given sample without any a priori information, we aimed at exploring the conditions of preservation of the relics and possible conditions of death. Metaproteomics and high-resolution microtomography imaging approaches were combined. A dataset comprising 6731 high-resolution MS/MS spectra was acquired and 968 of these spectra could be assigned to specific peptidic biomolecules. Based on the taxonomical information encompassed by the identified peptide sequences, 5 phyla were identified amongst eukaryota (94% of the biomass): Ascomycota (55%), with the species Aspergillus versicolor, Trichophyton mentagrophytes and Aspergillus glaucus, corresponding to expected cadaverous fungal flora; Chordata (42%), represented by a unique species, Homo sapiens; Streptophyta (3%); and Arthropoda (traces). Bacteria (6% of the biomass) were poorly represented. No trace of embalming substance could be retrieved, nor any pathogens. Imaging evidenced no heart defect nor embalming traces. No evidence that was inconsistent with natural and spontaneous conservation could be retrieved. This study prefigures the power of modern molecular techniques such as paleoproteotyping coupled to microtomography to gain insight into historical relics.

[The possibility of diagnosing the prescription of death in the study of embalmed corpses by the method of spin probes. (Experimental study)]. / Vozmozhnost' diagnostiki davnosti nastupleniya smerti pri issledovanii zabal'zamirovannykh trupov metodom spinovykh zondov. (Eksperimental'noe issledovanie).

THE AIM OF THE STUDY: Is to establish the possibility of studying the vitreous body of the eye from embalmed corpses using the spin probe method to develop diagnostic criteria for the prescription of death coming. The article presents data showing the possibility of diagnosing the prescription of death coming by the method of spin probes with the examination of the vitreous body of the eye from embalmed corpses. A total of 510 spectra obtained during incubation of the 1-oxyl-4-oxo-2.2.6.6-tetramethyl-piperidin spin probe in the vitreous body were studied. Measurements of each of the 6 objects were carried out daily for 10 days after embalming. The prospects of studying the vitreous body by spin probes in embalmed corpses for the development of diagnostic criteria for the prescription of death coming up to 4 days from the moment of embalming have been established.

Application of frozen Thiel-embalmed specimens for radiotherapy delineation guideline development: a method to create accurate MRI-enhanced CT datasets.

PURPOSE: Thiel embalming followed by freezing in the desired position and acquiring CT + MRI scans is expected to be the ideal approach to obtain accurate, enhanced CT data for delineation guideline development. The effect of Thiel embalming and freezing on MRI image quality is not known. This study evaluates the above-described process to obtain enhanced CT datasets, focusing on the integration of MRI data obtained from frozen, Thiel-embalmed specimens. METHODS: Three Thiel-embalmed specimens were frozen in prone crawl position and MRI scanning protocols were evaluated based on contrast detail and structural conformity between 3D renderings from corresponding structures, segmented on corresponding MRI and CT scans. The measurement error of the dataset registration procedure was also assessed. RESULTS: Scanning protocol T1 VIBE FS enabled swift differentiation of soft tissues based on contrast detail, even allowing a fully detailed segmentation of the brachial plexus. Structural conformity between the reconstructed structures on CT and MRI was excellent, with nerves and blood vessels imported into the CT scan never intersecting with the bones. The mean measurement error for the image registration procedure was consistently in the submillimeter range (range 0.77-0.94 mm). CONCLUSION: Based on the excellent MRI image quality and the submillimeter error margin, the procedure of scanning frozen Thiel-embalmed specimens in the treatment position to obtain enhanced CT scans is recommended. The procedure can be used to support the postulation of delineation guidelines, or for training deep learning algorithms, considering automated segmentations.

Thirty years of Thiel embalming-A systematic review on its utility in medical research.

In 1992, Walter Thiel described and embalming method that rendered "lifelike" tissues. Over the last 30 years, the Thiel method has been introduced worldwide for medical training and scientific purposes. This review examines research which can be linked to the use of Thiel embalming. A systematic review was performed to identify articles published in the following categories: research content, disciplines involved, sources and quantities of tissues deployed, and changes in research scope related to changes in the chemical composition of Thiel embalming. Four-hundred twenty-four publications were included. A number of adaptations to the original Thiel protocol were found, aiming to provide suitable tissue-substitutes in the development of emerging medical technologies or procedures. Musculoskeletal surgery, anesthesia and intensive care were the most common disciplines that used Thiel embalmed tissues for research. Anatomy and biomechanics played a lesser role. An increase over time was observed in research outputs related to the Thiel method, while the number of specimens used per study decreased. The main centers using Thiel embalming were in Graz, Dundee, Sapporo, Bern, Zurich and Ghent, which jointly accounted for more than 54% of all research conducted using this method. Following three decades of use, the Thiel method has evolved into being a well-established embalming technique for research purposes. Its future is challenged by the demanding requirements on both technical facilities and personnel, limitations of certain chemicals for use as fixatives, costs, and questions as to how "lifelike" the embalmed-tissues are from an objective standpoint, all of which warrants future investigations.

Needle insertion forces and fluid injection pressures during targeting of nerves in a soft embalmed cadaver model.

PURPOSE: Forceful needle-nerve contact and high subepineural pressures and are recognised causes of nerve damage. Pressure and force measurements are necessary to inform the mechanisms of nerve injury, build virtual simulator environments and provide operator feedback during simulation training. However, the range of pressures and forces encountered at tissue layers during targeted needle insertion and fluid injection are not known. METHODS: We built a needle that recorded in-line pressure during fluid injection and continuously measured force at the needle tip. Two anaesthetists were randomised to insert a 21 g block needle at 48 nerve sites on both sides of 3 soft embalmed Thiel cadavers. Our objective was to measure pressure and force during the course of targeted nerve injection at epimysium, in perineural tissue, on epineurium and during subepineural injection. At each interface, we infused a 0.5 ml bolus of embalming solution at a rate of 12 ml min-1 and recorded the pressure response. Force was measured continuously in the background throughout the procedure. RESULTS: Pressure was greater at epineurium and within subepineurium than perineural tissue, geometric ratio (95% CI) 4.7 (3.0-7.3) kPa and 3.8 (2.5-5.7) kPa, respectively, both P < 0.0001. Force on nerve contact and on nerve penetration was greater than force in perineural tissue, geometric ratios (95% CI) 3.0 (1.9-4.7) N and 3.6 (2.2-7.5) N, respectively, both P < 0.0001. On nerve contact, 1 in 6 insertions were ≥ 5 N CONCLUSIONS: Despite valid infusion pressures, anaesthetists exerted excessive force on nerves.

Non-Invasive Paleo-Metabolomics and Paleo-Proteomics Analyses Reveal the Complex Funerary Treatment of the Early 18th Dynasty Dignitary NEBIRI (QV30).

Biochemical investigations were carried out on the embalmed head of Nebiri (Museo Egizio, Turin; S-5109)-an 18th Dynasty Ancient Egyptian dignitary-and on the canopic jar containing his lungs (Museo Egizio, Turin; S. 5111/02) with the aim of characterizing the organ's (lung) specific paleo-proteins and of identifying the compounds used in his embalming "recipe". The application of a functionalized film method allowed us to perform a non-invasive sampling. Paleo-proteomics confirmed the presence of lung tissue-specific proteins (organ specific) as well as the presence of proteins linked to severe inflammation. Paleoproteomics and paleometabolomics further allowed the identification of the main components of Nebiri's embalming recipe: animal fats and glue, balms, essential oils, aromatic plants, heated Pistacia, and coniferous resins. Both the use of Pistacia and coniferous resins in an early 18th Dynasty individual confirm Nebiri's high social status. The technique applied offers a targeted approach to the chemical characterization of human tissues, embalming compounds, and organic materials layering in pottery. The ability of the functionalized film method to harvest all types of compounds, from macromolecules (i.e., proteins) to small molecules (i.e., organic acids) opens a new path in the study of ancient material culture; furthermore, it allows to perform untargeted analysis, which is necessary when no a priori information is available.

Rethinking the Process of Animal Mummification in Ancient Egypt: Molecular Characterization of Embalming Material and the Use of Brassicaceae Seed Oil in the Mummification of Gazelle Mummies from Kom Mereh, Egypt.

The study of animal mummification in ancient Egypt has recently received increasing attention from a number of modern scholars given the fact that this part of ancient Egyptian funerary and religious history is a practice yet to be fully understood. In this study, nine samples of embalming matter were extracted from six gazelle mummies from the archaeological site of Kom Mereh (modern village of Komir), dated to the Roman period of dominance in ancient Egypt. All samples were analyzed for the presence of inorganic and organic matter applying a multi-analytical approach based on Fourier transform infrared spectroscopy (FT-IR) and gas chromatography-mass spectrometry (GC-MS). Furthermore, in order to identify more specific compounds such as bitumen and beeswax in studied balms, each sample was subjected to a solid phase extraction (SPE) and saponification separation process, respectively. The results of this study revealed that the majority of the analyzed embalming substances sampled from six gazelle mummies from Kom Mereh were complex mixtures of plant oils, animal fats, conifer resin, and beeswax. In this regard, this study was able to report a practice until now unmentioned in the scientific literature, namely, the use of cruciferous oil, derived from seeds of Brassicaceae plants, in animal mummification.

The advantages of modified Thiel technique in head and neck surgical anatomy teaching.

PURPOSE: Thiel's body preservation method allows the donor body to retain color and soft tissue that are similar to those of a living individual. Since its initial description, the technique has not been developed much. Here, we propose a simpler protocol applied to the head and neck, making the technique easier, cheaper, and more accessible to a greater number of medical schools. METHODS: The modified Thiel technique was applied to three heads separated from the body, followed by 6 h of perfusion and 6 weeks of fixation. This technique was compared with formalin (three heads) and freeze (three heads) preservations during academic training in head and neck surgery. Anatomical dissections included a parotid gland dissection, a submandibular gland dissection, an otologic and an endonasal dissection. Twelve surgeons blindly assessed the three types of preservations using a standardized questionnaire. RESULTS: The modified Thiel technique made possible better quality of dissection and tissue identification, with the exception of endonasal dissection. Concerning the endonasal dissection, the modified Thiel technique ranked best for smell, but all other criteria ranked lower than the freezing method. For the submandibular and parotid gland dissections, the modified Thiel technique was ranked best, with statistically significant differences (p < 0.002) for all items. The modified Thiel technique also ranked significantly better during otologic surgery regarding the quality of the skin/eardrum, bone, and muscle, and the smell. CONCLUSION: This study demonstrated that the modified Thiel technique is an embalming technique which improves the quality of head and neck surgical anatomy education.

Contributions of the Thiel's Method in teaching and researching anatomy.

INTRODUCTION: Knowledge of the human body is based on teaching and research. Anatomy remains an essential prerequisite for medical personnel to know the human body. The idea of Thiel's technique is to preserve the natural texture, volume, color and shape of the body. The objective of this study was to analyze a literature review of this technique from the perspective of anatomy teaching and biomechanical research. METHOD: We have taken up the process of Thiel's method from the original publication, stating the known applications of this technique in teaching and research. We have integrated into our study the specific advantages of using bodies preserved by this method in intertropical countries with a warm climate: the example of Gabon. DISCUSSION: Biomechanical research and teaching of medical and surgical sciences are regularly performed on human cadavers. Anatomical dissection therefore represents one of the main activities of anatomy laboratories. We have limited our analysis to aspects of anatomy teaching, research, and clinical and surgical practice. CONCLUSION: We have sought to popularize the Thiel body preservation technique. It offers many advantages. Teaching and research on human cadavers preserved by this method is an educational alternative. The simulation centre creates the conditions of a surgical block for the learners. We recommend this technique to anatomists and clinicians. This technique seems to be very interesting for structures with a limited number of bodies.

Neutralisation of SARS-CoV-2 by anatomical embalming solutions.

Teaching and learning anatomy by using human cadaveric specimens has been a foundation of medical and biomedical teaching for hundreds of years. Therefore, the majority of institutions that teach topographical anatomy rely on body donation programmes to provide specimens for both undergraduate and postgraduate teaching of gross anatomy. The COVID-19 pandemic has posed an unprecedented challenge to anatomy teaching because of the suspension of donor acceptance at most institutions. This was largely due to concerns about the potential transmissibility of the SARS-CoV-2 virus and the absence of data about the ability of embalming solutions to neutralise the virus. Twenty embalming solutions commonly used in institutions in the United Kingdom and Ireland were tested for their ability to neutralise SARS-CoV-2, using an established cytotoxicity assay. All embalming solutions tested neutralised SARS-CoV-2, with the majority of solutions being effective at high-working dilutions. These results suggest that successful embalming with the tested solutions can neutralise the SARS-CoV-2 virus, thereby facilitating the safe resumption of body donation programmes and cadaveric anatomy teaching.

Phonosurgery Training in Human Larynx Preserved with Thiel's Embalming Method.

BACKGROUND: To describe the suitability of larynges preserved with Thiel's embalming method for phonosurgery training. METHODS: A training model for phonosurgery techniques simulating vocal pathology and glottal insufficiency is developed to compare and evaluate the perception of embalmed vocal cords through a voluntary and anonymous survey rated on a scale of 1-5. A total of 10 residents and young otolaryngologists participated in the surgical training in phonosurgery. RESULTS: Ten larynges preserved in formalin and 10 Thiel's embalmed larynges were used for the investigation. Phonosurgery procedures were performed following microflap and injection laryngoplasty techniques. The larynges preserved with Thiel's method demonstrated vocal cords that maintain their pliability and good tissue quality allowing a sensation of realism compared to the living body and providing suitable conditions for realistic laryngeal training. Participants held a positive experience, believed them to be useful and that these models of embalmed larynges were similar to the clinical setting and improved skills and confidence in performing phonosurgery. CONCLUSIONS: The human larynges embalmed with Thiel's method maintain the pliability of the vocal cords, thus representing a unique model to practice and reproduce training for endolaryngeal procedures without the risks of contamination, anatomical variation, or rigidity of other models.

Thiel-embalmed cadavers as a novel training model for ultrasound-guided supine endoscopic combined intrarenal surgery.

OBJECTIVE: To evaluate Thiel-embalmed cadavers (TEC) as a training model for percutaneous nephrolithotomy (PCNL), more specifically for ultrasound (US)-guided supine endoscopic combined intrarenal surgery (ECIRS). SUBJECTS AND METHODS: Thirteen urologists (nine experienced endourologists, four fellows/residents) performed an US-guided supine ECIRS procedure on a TEC. The model was evaluated by way of a questionnaire (5-point Likert scale). Descriptive statistical analysis was performed and results were graphically presented using divergent bar graphs. RESULTS: US images were appreciated as lifelike in all aspects. Although distention of the collecting system was not ideal in one out of three TEC, US visualisation of the distended calyces during puncture was good. Skin penetration was more difficult and less realistic in TEC, while kidney puncture and dilatation were deemed very realistic. Ureteric and collecting system anatomy and consistency were similar to real life, although the mucosa appeared paler. US needle guidance was perceived as excellent. Overall, needle puncture assessment was realistic and useful as a training tool. Overall quality and satisfaction of TEC in US-guided supine ECIRS was good to excellent and comparable to a real-life procedure. Overall appropriateness of the TEC model was considered excellent for both initial and advanced supine PCNL training. CONCLUSION: Despite the minor drawbacks of tough skin and non-ideal collecting system dilatation during ureteroscopy, the TEC model was considered good to excellent as a training model for US-guided PCNL, in particular, US-guided needle puncture of the kidney during supine ECIRS.

An ideal solution? Optimising pretreatment methods for artificially mummified ancient Egyptian tissues.

RATIONALE: Although the analysis of skeletal remains dominates the study of ancient dietary stable isotopes, mummified bodies also allow short-term diet to be studied through the analysis of soft tissues. The application of resins, waxes and oils during mummification can affect the results obtained. This study assesses a range of methods for removing such substances from mummified tissue. METHODS: An experimental mummification model following ancient Egyptian methods was created using a modern pig leg. Sub-samples of skin, muscle and bone were removed and coated with a range of substances used in Egyptian mummification. Four methods were used to clean these samples before the measurement of the carbon and nitrogen stable isotope ratios of their gelatinised collagen content using a ThermoFinnigan Flash Elemental analyser coupled to a DeltaPlus XL isotope ratio mass spectrometer via a ConFlo III interface. RESULTS: The results showed that embalming materials can significantly affect dietary stable isotope ratios, and that these substances are most effectively removed using a mixture of polar and non-polar solvents. Results indicate that bone samples demineralised with HCl and skin samples produce more accurate results than bone samples demineralised with EDTA or muscle samples. CONCLUSIONS: The choice of tissue and the preparation methods used can have a significant effect on the accuracy of stable isotope data obtained from mummified tissue, particularly when embalming materials are also present. A mixture of solvents appears to be a more effective cleaning agent than a single solvent. Demineralisation with HCl is preferable for well-preserved bone, as used in this study, but whether this is the case for more fragile, less well-preserved bone requires further study. Skin samples produce more consistent data than muscle, but visually distinguishing between these tissues is not simple on ancient mummies.

UV-C Irradiation Reduces the Experimentally Induced Bacterial Load on the Surface of a Human Cadaver: An Additional Option for the Preservation of Cadavers in Anatomy.

Safety is a major issue in the embalming procedures of human cadavers. Reduced application of formaldehyde is often recommended. The aim of this study was to investigate the potency of ultraviolet light (UV-C irradiation) on the bacterial load on the surface of a conserved human cadaver. To test UV-C irradiation, the cadaver was laid out in the dissection hall and, after preparation of the muscles, was covered with linen sheets moistened with water. Swabs of the surface and microbiological analysis revealed sporadic bacterial colonies. The surface area was then spiked with bacteria and irradiated by a UV lamp for 15 or 60 min. Half of the area was covered by aluminum foil to serve as a control. After exposition, swabs were taken and analyzed. The exposition had reduced the number of colonies to one third (15 min exposition) and to one tenth (60 min exposition) of the control area. Thus, UV-C irradiation could be used in the preservation of cadavers without chemical pollution of the environment and without any risk for the employees. Clin. Anat. 32:113-116, 2019. © 2019 Wiley Periodicals, Inc.

Flammability and combustion hazard of preserved human tissues.

Preservation techniques have evolved over the years to respond to the need of longer dissection periods, with formaldehyde being widely used for this purpose. In recent years, efforts have been focusing on reducing the health hazards of this fixative and the rigidity of the tissues embalmed with it. With every embalming technique that they are either developing or resurrecting from various protocols, institutions need to assess the fire and health hazards of all the chemicals being used. Compliance with the storage and handling safeguards listed in the Material Safety Data Sheet of each chemical, needs to be accompanied by infrastructure changes. To reduce the health hazards of formaldehyde, institutions are taking appropriate countermeasures directed at the source itself, by using substitutes or injecting formaldehyde chelating agents, and are adopting high performance air extraction systems to protect the users. However, little is known about the flammability risk of embalmed human bodies. During a recent visit to McGill University morgue by architects and safety inspectors to expand it, we were asked about the flammability risk of the embalmed specimens, so we carried out a flashpoint test to evaluate the flammability potential of specimens embalmed with each of our three embalming techniques: formaldehyde-based, Thiel, and phenol-based. With the phenol-based embalming showing the most risks, attention should be drawn to all institutions who are experimenting with various embalming techniques to evaluate the flammability potential of their embalmed specimens.