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Subcutaneous tissue from the Iceman “Ötzi.” (a) The well-preserved fat tissue is separated by a fresh bleeding zone. (b) More on detail, the bleeding zone shows on special staining (Prussian blue) the remnants of haemosiderin-positive macrophages (blue) indicating a survival period of several days ante mortem (original magnification: (a) ×150; (b) ×300; (a): H&E; (b): Prussian blue stain).
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The application of histology to soft tissue remains offers an important technique to obtain diagnostically important information on various physiological and pathological conditions in paleopathology. In a series of 29 cases with mummified tissue ranging between 16 months and c. 5.200 years of postmortem time interval, we systematically investigate...
Citations
... The sample was gently shaken for 12-16 hours to ensure optimal penetration of the solution into the material. After this period the solution was replaced by 4-6% formaldehyde, pH 7.4 without any further addition for another 12-16 hours (postfixation) (Grove et al., 2015). Then the sample was decalcified in Osteodec solution (Bio-Optica) and it was subsequently processed for paraffin embedding and sectioned to a thickness of 3-5 µm. ...
Although historically widespread in most of the shallow and warm waters of the world, the presence of sawfishes in the Mediterranean Sea is still a debated question. For some authors, they never inhabited this basin except as vagrants, while for other authors both Pristis pristis and Pristis pectinata were present in the Mediterranean Sea but were extirpated in the 1960s–1970s. The sawfish rostrum kept in the Basilica Santuario del Carmine Maggiore (Naples, Italy), and considered by some authors the first record of sawfish in the Mediterranean was studied using a combination of morphological, genetic, radiocarbon dating and histochemical staining methods to validate the taxonomic identification, estimate its age and assess its geographical origin. Results indicate that the rostrum does not belong to P. pristis as reported by previous authors, but instead possesses morphological and genetic characters typical of P. pectinata. In addition, the radiocarbon age shows that the rostrum is more recent than previously believed, dating it back to the mid-nineteenth century, and genetic and meristic results cast doubt on its presumed Mediterranean origin. This study demonstrates that historical records should always be critically evaluated before using them to draw any far-reaching conclusion about species’ past ecology and/or biogeography, and that future studies using historical information and specimens should adopt an integrative taxonomy approach similar to the one used here.
... The histological analysis was prepared with a rehydration procedure, followed by the embedding and cutting as previously described in detail (14). The prepared sample was serially . ...
... /fmed. . sectioned (2-3 µm thickness) and then stained with H&E, connective tissue stains, PAS, Grockotts' silver stain, and Prussian blue (14). ...
We describe here the results of a multidisciplinary study on an infant mummy from 16th century Upper Austria buried in the crypt of the family of the Counts of Starhemberg. The macroscopic-anthropological, radiological (whole-body CT scan), histological (skin tissue), and radiocarbon isotope investigations suggested a male infant of 10–18 months' age, most likely dying between 1550 and 1635 CE (probably Reichard Wilhelm, 1625–1626 CE), that presented with evidence of metabolic bone disease with significant bilateral flaring of costochondral joints resembling “rachitic rosary” of the ribs, along with straight long bones and lack of fractures or subperiosteal bleeding residues. Although incompletely developed, the osteopathology points toward rickets, without upper or lower extremities long bone deformation. The differential diagnosis is vitamin C deficiency (scurvy) (also with an incomplete presentation, although overlap between both disorders may be present). As additional pathology, there was significantly enlarged subcutaneous fat tissue (thickness more than 1 cm at the navel and thighs and longitudinal creases of the skin) along with a histologically enlarged subcutaneous fat layer consistent with infantile adipositas as a coincident disorder. Finally, remnants of lung tissue with pleural adhesion of the right lung indicate possibly lethal pneumonia, a disease with an increased prevalence in vitamin D deficient infants. Ultimately, the skull presented with extensive destruction of the bones of the base and dislocation of the bones of the skull squama. These changes, however, are most likely post-mortal pseudopathology, the result of a burial in a flat, narrow coffin because there were no bone fractures or residues of bleeding/tissue reaction that would have occurred whilst the patient was alive.
... Specimens obtained from several organs and tissues were processed for histological analysis. The preparation of the samples, the rehydration procedure, and the embedding and cutting techniques, have been previously described in detail [16]. Particularly, samples from both lungs (especially from areas with obvious calcification, but also from all different lobes), the heart, and the abdominal structures, were prepared. ...
... All histological samples were stained with H&E, connective tissue stains, PAS, Grockotts silver stain, Prussian blue and May-Grünwald-Giemsa stain, Ziehl-Neelsen acid fast bacilli stain, and Kongo red staining for amyloid deposition [16]. ...
... DNA was extracted and amplified by PCR as done in established protocols [21]. The resulting aDNA was then used to identify the specific sequence of the MTC, IS6110, as performed previously [16] by gel electrophoresis and subsequent Sanger sequencing. Finally, further aliquots were used to identify substrains of the MTC by using the spoligotyping technique as previously described [21]. ...
In paleopathology, morphological and molecular evidence for infection by mycobacteria of the M. tuberculosis complex (MTC) is frequently associated with early death. In the present report, we describe a multidisciplinary study of a well-preserved mummy from Napoleonic times with a long-standing tuberculous infection by M. tuberculosis senso stricto who died at the age of 88 years of focal and non-MTB related bronchopneumonia. The well-preserved natural mummy of the Royal Bavarian General, Count Heinrich LII Reuss-Köstritz (1763-1851 CE), was extensively investigated by macro- and histomorphology, whole body CT scans and organ radiography, various molecular tissue analyses, including stable isotope analysis and molecular genetic tests. We identified signs for a long-standing, but terminally inactive pulmonary tuberculosis, tuberculous destruction of the second lumbar vertebral body, and a large tuberculous abscess in the right (retroperitoneal) psoas region (a cold abscess). This cold abscess harboured an active tuberculous infection as evidenced by histological and molecular tests. Radiological and histological analysis further revealed extensive arteriosclerosis with (non-obliterating) coronary and significant carotid arteriosclerosis, healthy bone tissue without evidence of age-related osteopenia, evidence for diffuse idiopathic skeletal hyperostosis and mild osteoarthrosis of few joints. This suggests excellent living conditions correlating well with his diet indicated by stable isotope results and literary evidence. Despite the clear evidence of a tuberculous cold abscess with bacterioscopic and molecular proof for a persisting MTC infection of a human-type M. tuberculosis strain, we can exclude the chronic MTC infection as cause of death. The detection of MTC in historic individuals should therefore be interpreted with great caution and include further data, such as their nutritional status.
... The comparison between male and female was found to be non-significant (p<0.05) when season of recovery were compared. The time between death and postmortem varies from [6][7][8][9][10][11][12][13] , and 15 were 2 (8%), while on each days 9 and 11 the bodies recovered were 3 (12%). The chi square did not show any significant difference within the days (p>0.05). ...
... Our study showed that most of the cases recovered in the month of April to July had adipocere formation in short time when the weather was hot and humid, the average room temperature ranged between 27-39°C with humidity between 20-90%. Our results are in consistent with Sikary et al 9 who showed adipocere formation in hot humid climatein short period and declared that in hot and humid climate external water or moisture is not needed and the water present in the decomposing tissue is sufficient for adipocere formation 11,12 . Same results were seen by Stuart et al 7 who showed more than half of the bodies in each vault had extensive adipocere formation in very hot humid climate within days and agreed that only internal body water is sufficient for adipocere formation and even moisture was not necessary. ...
... Adipocere is favored by moisture, warm, anaerobic environment and personal characteristics, while inhibited in cold, and aerobic environment. It is more frequently seen in obese, females and well-nourished newborn infants 9,11 . Adipocere helps in determining injury pattern, identification, cause of death and time since death 2-3 . ...
... The comparison between male and female was found to be non-significant (p<0.05) when season of recovery were compared. The time between death and postmortem varies from [6][7][8][9][10][11][12][13] , and 15 were 2 (8%), while on each days 9 and 11 the bodies recovered were 3 (12%). The chi square did not show any significant difference within the days (p>0.05). ...
... Our study showed that most of the cases recovered in the month of April to July had adipocere formation in short time when the weather was hot and humid, the average room temperature ranged between 27-39°C with humidity between 20-90%. Our results are in consistent with Sikary et al 9 who showed adipocere formation in hot humid climatein short period and declared that in hot and humid climate external water or moisture is not needed and the water present in the decomposing tissue is sufficient for adipocere formation 11,12 . Same results were seen by Stuart et al 7 who showed more than half of the bodies in each vault had extensive adipocere formation in very hot humid climate within days and agreed that only internal body water is sufficient for adipocere formation and even moisture was not necessary. ...
... Adipocere is favored by moisture, warm, anaerobic environment and personal characteristics, while inhibited in cold, and aerobic environment. It is more frequently seen in obese, females and well-nourished newborn infants 9,11 . Adipocere helps in determining injury pattern, identification, cause of death and time since death 2-3 . ...
Objective: To find out duration of adipocere formation in hot and humid area. Study Design: Cross sectional study. Place and Duration of Study: Mohi-Ud-Din Islamic Medical College Mirpur Azad Kashmir and HITEC-IMS Taxila, from Jan 2017 to Jan 2019. Methodology: Total 25 cases with adipocere formation were included in this study. The cause of death, time since death, death circumstances, surrounding environment from where the body was recovered and demographic profile of the deceased were considered, while cases of septicemic deaths were excluded from the study. The adipocere was confirmed by forensic medicine consultants by its soft, moist and greasy consistency, grayish white appearance and ammoniacal smell. Results: Most of the cases recovered during months of March to June from open and closed areas, while 3 cases were recovered from water source or river banks. The time duration of recovery from time of death was from 5 days to 15 days 3h. The adipocere formation showed a high odds ratio 2.0-(95%-CI-0.115-3.482) and OR-1.5-(95%-CI-.098-2.306) for bodies recovered from water and open areas respectively. The adipocere formation in hot and rainy season showed OR-3.3-(95%-CI .190-4.199) and OR-2.4-(95%-CI .122-3.642) respectively. The adipocere formation occurred more during days 1-5 with an OR-1.9-(95%-CI .221-1.625). Conclusion: The adipocere formation can be seen as early as 2 days in a hot humid environmentin closed areas, while the extent of adipocere is larger in bodies recover from water source.
... Once the soft tissue is obtained, a technical procedure similar to that used in conventional samples of fresh tissue can be followed: formalin fixation, paraffin embedding, microtome sectioning and histochemical staining. Nonetheless, all of this is only possible if we first return the tissue to a rehydrated state with the use of some simple solutions like those of Ruffer or Sandison, which are mainly based on an alcoholic solution with sodium carbonate [8,9,23,24]. ...
Paleopathology is a science located in a crossroad between history, archaeology, anthropology, and medicine an can offer unique historical knowledge by using techniques of traditional pathology as well as other branches of Medicine, which is especially fruitful when applied to ancient subjects in which soft tissues are preserved: mummies.
... The reasons remain unclear but could be due to the rapid decomposition of stomach tissue; therefore, it could be more difficult to identify the relevant gastric structures. As Grove and colleagues observed in their approach to the systematical analysis of soft tissue histopathology in palaeopathology, the brain and the kidney Pathobiology 2018;85:267-275 DOI: 10.1159/000490797 have poor conservation potential and are more difficult to identify due to the lack of connective tissue and absence of certain "key structures," which is instead the case of lung and liver tissues [65]. ...
Ancient Egyptian human remains have been of interest in the fields of both medical and Egyptological research for decades. However, canopic jar holders for internal organs (liver, lungs, stomach, intestines) of Egyptian mummies appear to be but a very occasional source of data for such investigations. The few medical approaches focusing on the content of these jars are summarized and listed according to pathogens and diseases to give a structured overview of this field of study. An extensive search of the literature has been conducted from different bibliographic databases with a total of n = 26 studies found. The majority of diseases found consisted of infectious diseases and internal medicine conditions such as schistosomiasis or emphysema. These are just 2 examples of many that, instead of primarily affecting bone, muscle or skin, specifically target internal organs. Hence, a better understanding of the evolution of diseases that still affect mankind could be gained. In conclusion, this reassessment shows that canopic jars represent a highly underestimated source for histological, radiological and ancient DNA examination of Ancient Egyptian remains and should, thus, be more and more brought back into the focus of retrospective medical research.
... These biopsy tissue on living or necropsies on the non-living. In all histological operation, good preservation of tissue and/or cellular constituents remains essential [2,3]. ...
... Generally, the tissue is without any remaining cells or soft tissues; so-called skeletonized or "dry" bone tissue. Sometimes mummification allows for the analysis of preserved soft tissue (Aufderheide, 1981;Hess et al., 1998;Ciranni et al., 2000;Grove et al., 2015) but such findings are relatively rare. The remainder of this article will thus focus on dry bone tissue. ...
Paleopathology is the study of trauma and disease as may be observed in ancient (human) remains. In contrast to its central role in current medical practice, microscopy plays a rather modest role in paleopathology. This is at least partially due to the differences between fresh and decomposed (i.e. skeletonized or ‘dry bone') tissue samples. This review discusses these differences and describes how they affect the histological analysis of paleopathological specimens. First, we provide a summary of some general challenges related to the histological analysis of palaeopathological specimen. Second, the reader is introduced in bone tissue histology and bone tissue dynamics. The remainder of the paper is dedicated to the diagnostic value of dry bone histology. Its value and limitations is illustrated by comparing several well-studied paleopathological cases with similar contemporary, clinical cases. This review illustrates that due to post-mortem loss of soft tissue, a limited number of disorders display pathognomonic features as may be observed during histological analysis of skeletonized human remains. In the remainder of cases, histology may help to narrow down the differential diagnosis or is diagnostically unspecific. A comprehensive, multidisciplinary diagnostic approach therefore remains essential in the paleopathological diagnostic process. This article is protected by copyright. All rights reserved.
The combination of multi-omic techniques, e.g. genomics, transcriptomics, proteomics, metabolomics and epigenomics has revolutionised studies in medical research. These are employed to support biomarker discovery, better understand molecular pathways and identify novel drug targets. Despite concerted efforts in integrating omic datasets, there is an absence for the integration of all four biomolecules in a single extraction protocol. Here, we demonstrate for the first time a novel, minimally destructive integrated protocol for the simultaneous extraction of artificially degraded DNA, proteins, lipids and metabolites from pig brain samples. We used an MTBE-based approach to separate lipids and metabolites, followed by subsequent isolation of DNA and proteins. We have validated this protocol against standalone extraction protocols and show comparable or higher yield of all four biomolecules. This integrated protocol is key towards facilitating preservation of irreplaceable samples while promoting downstream analyses and successful data integration by removing bias from univariate dataset noise and varied distribution characteristics.
