Paleopathology in South American mummies: a review and new findings

This article is a review of research and additional unpublished diseases that have been discovered and documented in naturally mummified remains recovered from South America. A new impetus in paleopathological studies was the work and discovery of a solution for rehydration of mummified tissues by Sir Marc Armand Ruffer in 1913. This solution allows the paleopathologist, after performing the autopsy, to process the tissues in a manner similar to current practices in pathology. In our studies, the most common diseases were infectious in nature, similar to the diseases that are most prominent today in the same regions.     

Histological, histochemical, enzyme histochemical and ultrastructural investigations of the testis of Padogobius martensi between annual breeding seasons

From July to March, the testis of the spring‐spawning freshwater goby Padogobius martensi is characterized by spermatogonial proliferation. A close correlation exists among type of proliferating spermatogonia, gonado‐somatic (I_G) profiles and morphological and functional variations of the Leydig cells. The I_G reach their minimal levels by the end of summer and increase progressively but modestly during autumn and winter. Declining I_G levels are associated with proliferation of primary spermatogonia only, whereas increasing I_G levels are associated with predominant proliferation of secondary spermatogonia. Minimal I_G levels are reached when the germinal epithelium is formed by a continuum of primary spermatogonia and associated Sertoli cells. The proliferation of secondary spermatogonia begins only at this time. Spermatogenesis in autumn occurs when spermatogonial cysts contain at the most 16 cells and it rarely results in the maturation of several cysts so that the amount of sperm cells produced is either negligible or scarce. A number of degenerating cells are usually present within the spermatogonial and meiotic cysts. Leydig cells are the unique cells that display features of steroidogenic cells: mitochondria with tubular cristae, extensive smooth endoplasmic reticulum (SER), 3β‐hydroxysteroid dehydrogenase (3β‐HSD) and glucose‐6‐phosphate dehydrogenase (G6PD) activity and sudanophilia. Light and dark Leydig cell varieties are always present. During regression, Leydig cells undergo a marked decrease in SER amount, mitochondrial sizes and number of mitochondrial cristae. In parallel, the 3β‐HSD and G6PD activities and sudanophilia decrease progressively until they become undetectable by the end of regression. In autumn, mitochondria increase in size, reaching sizes similar to those observed at the end of the spawning season in the light cells, but not in the dark cells. The SER, on the contrary, undergoes a modest and irregular increase only in a part of the Leydig cells, mostly of the light type. In parallel, the 3β‐HSD and G6PD activities increase until they become moderately intense by the end of autumn. At the end of winter, the SER is extensive and regularly dilated in both Leydig cell types, whereas mitochondria still have sizes similar to those observed in December. The 3β‐HSD and G6PD activities are strong and sudanophilia is again detectable. Sertoli cells undergo changes in shape and position in relation to the proliferation of primary spermatogonia and the development of cysts. A junction modulation occurs in association with these changes. Sertoli cells also undergo changes indicative of a decrease in activity immediately after spawning (loss of mitochondrial cristae and clarification of the mitochondrial matrix) and of an increase in activity by the end of the regressing phase (darkening of the mitochondrial matrix and increase in mitochondrial cristae, rough endoplasmic reticulum (RER) and free ribosomes). In addition, they are involved in the phagocytosis of degenerating germ cells at all stages of their development. Macrophages are found in the testis interstitium only, where they are usually adjacent to Leydig cells, myoid cells and blood capillaries and do not participate in the phagocytosis of degenerating germ cells. Myoid cells do not undergo ultrastructural changes except for an increase in the amount of heterochromatin by the end of spawning. The meaning of the autumnal spermatogenic wave and the relationships between the development of the germinal epithelium and the changes of the Leydig and Sertoli cells are discussed.     

Histological, histochemical, and fine structural observations on the spleen of seals

Spleens of three species of Antarctic seals with different diving habits (Weddell seal, crabeater seal, and fur seal) have been studied with histological, histochemical, and electron microscopic methods. The spleens can be classified as nonsinusoidal, with capsule and trabeculae rich in innervated smooth muscle cells. The trabecular system is particularly well developed in the deep- and long-diving Weddell seal. As in other mammals the pulp can be divided into white and red pulp. In the white pulp, periarteriolar lymphatic sheaths and secondary lymphatic nodules occur; both are surrounded by a marginal zone rich in macrophages and eosinophils. The nodules can be observed frequently, which is in accordance with abundance of plasma cells in the red pulp. Well-developed white pulp and numerous plasma cells and eosinophils obviously reflect a high load of nematodes, which have mainly been found in lung and stomach. Additionally, in the red pulp morphological evidence for the following functions has been found: destruction of erythrocytes, erythropoiesis, and thrombopoiesis. In respect to blood flow through the red pulp, we interpret our observations in the following way: terminal branches of arterioles open into the space between the fibroblastic reticulum cells; blood draining from here is collected into pulp veins, which are mainly found near the trabeculae. Thus, the seals have an open vascular compartment in their spleens, as also occurs in the cat. The red pulp is innervated by numerous nerve fibers that seem to include both cholinergic and adrenergic ones. The target cells of these fibers seem to be the fibroblastic reticulum cells, whose state of contraction may influence the direction of blood flow through the red pulp.     

Histological and histochemical studies on the formation of statoblasts of a fresh-water bryozoan,Pectinatella gelatinosa

The course of the statoblast formation in Pectinatella gelatinosa was divided into four stages and studied histologically and histochemically. The bottom of the cystigenous cup is a center of cystigenous cell differentiation and the peripheral zone of the inner cystigenous layer turns to the outer cystigenous layer as the cystigenous cup grows. The annulus is formed by migration and transformation of the outer cystigenous cells. During early stages, the yolk cells have an intensely pyroninophilic or RNA-rich cytoplasm. The cytoplasmic pyroninophilia then diminishes as the amount of yolk granules increases. Several kinds of yolk substances occur in the mature statoblast. During statoblast formation glycogen appears first, then glycoprotein and finally neutral unsaturated lipid. Acid phosphatase activity is associated with granular structures in the cytoplasm. In the cystigenous vesicle, acid phosphatase activity is low and confined to the apical extremity of the cell. Histochemically detectable alkaline phosphatase activity is not involved in the formation of the statoblast.     

Histological and histochemical techniques applied to the acaricales

Summary Schedules and technical methods in the preparation of permanently stained slides of members of the Agaricales are given. The results obtained from the examination of sections of members of the Bolbitiaceae and Boletaceae so prepared are discussed. The work described is that originally carried out by the late Dr.Max Barrett.     

Histological and histochemical characteristics of the bovine notochord.

In 20 bovine embryos and fetuses 6-65 mm long (crown-rump length) and 23 to 60-70 days old, the structure and localization of acid and neutral mucopolysaccharides and glycogen in their notochord were investigated. Also, the localization in the notochord was examined of the activity of alkaline and acid phosphatases, alpha-glycerophosphate-, glucose-6-phosphate-, isocitrate-, glutamate-, lactate- and succinic- dehydrogenases, and nicotinamide-adenine-dinucleotide- and nicotinamide-adenine-dinucleotide-phosphate- diaphorases. It was found that the bovine notochord begins decomposing at the end of embryonal and the beginning of fetal development (45-50 days old) and that in the fetus aged 55-65 days it no longer represents an unbroken cord of notochordal cells. Secretory activity of notochordal cells which produce the notochordal sheath starts very early (in 10 mm-long embryos), and interruptedly increases up to the end of the embryonal developmental period when regression appears at the beginning of the fetal period. These findings agree with findings in the human embryo where, however, they relate to earlier developmental periods.     

Evidence based palaeopathology: Meta-analysis of Pubmed-listed scientific studies on pre-Columbian,South American mummies

The aim of this study was to review all Pubmed^®-listed palaeopathological studies performed on pre-Columbian South American mummies. A total of 61 studies were found (1977–2005). Review criteria included e.g. method of examination, method of mummification, palaeopathological diagnoses and individual age of mummies as well as dating, which ranged from 7500 to 500 years BP, mainly (if reported) originating from the Chiribaya and Chinchorro cultures. The average age of the 99 individually reported mummies was about 25 years. Only six studies included computed tomography, thirteen studies used classical radiography as an examination method. Three studies analysed parasite related diseases, especially caused by Trypanosoma cruzi. Among all of the reported infectious diseases (n = 9), there were seven studies presenting cases of tuberculosis. The results were also compared inter-culturally. In 61 studies (37 epidemiological and 24 case reports) more than 6400 mummified individuals were analysed. By contrast, meta-analytic data for ancient Egyptian mummies ( Zweifel et al., 2009) included about 3000 analysed individuals in 131 studies (85 case reports and 46 epidemiological studies). In general, ancient Egyptian mummies were shown to be intentionally mummified, whereas the Pre-Columbian American mummies showed a great diversity of spontaneous mummification. However, ritualistic mummification methods were also practised (n = 2). This study's results shall assist to improve evidence-based research in palaeopathology.     

Histological and histochemical observations on the testis of Gobius paganellus

Summary Histological and histochemical observations on the testis of Gobius paganellus during all seasons of the year are described. In the yearly reproductive cycle, spawning in the Gulf of Naples occured from June through August, testicular recovery and relative inactivity from September through December, and active spermatogenesis from January to May.Germ cells develop as clones from single primary spermatogonia, each clone in a follicle enveloped by cells which are interpreted as Sertoli cell homologues.Glandular tissue is present in large amounts both in the form of interstitial islets and as a large mass along the length of the mesorchium. Cholesterolpositive lipids and ⁵-3-hydroxysteroid dehydrogenase, presumptive evidence for steroid production, are present exclusively in this tissue.Lipids are present in the glandular cells in acidic and neutral forms. The acid fats are the more abundant, but neutral lipids increase in amount in the period April-June. There is, furthermore, a cyclic variation in lipid droplet size, small droplets being present in the spring (during active spermatogenesis) and fewer, larger droplets during the fall (post-spawning period). Phospholipids are lacking.Lactic dehydrogenase activity was weak in the relatively inactive postspawning period, but was much more intense during active spermatogenesis.The glandular tissue in the testis of Gobius is interpreted as homologous with the interstitial (Leydig) tissue found in the testes of higher vertebrates.This investigation was supported by research grant RG-6455 from the Division of General Medical Sciences, U.S. Public Health Service.Postdoctoral Fellow from the Division of General Medical Sciences, U.S. Public Health Service. Supported in part by U.S. Public Health Service Training Grant 5 Tl GM-136, Department of Biological Structure, University of Washington, School of Medicine, Seattle, Washington, U.S.A.     

Histological examination of experimentally mummified tissues

Tissue mummified by desiccation can be examined histologically after rehydration with Ruffer's solution, and a number of natural mummies have been studied by this technique. In order to study the efficacy of the rehydration process, and to evaluate the degree of artefact introduced, normal human tissues were desiccated, rehydrated, and examined histologically. There was generally good preservation of architecture and moderate preservation of cellular detail, with some organ specific variability. The prospect of an atlas of mummified pathological lesions is discussed, as an aid to the determination of disease patterns in prehistoric populations.     

Histological and histochemical observations in the stomach of the Senegal sole, Solea senegalensis

An histological and histochemical study was conducted on the stomach of adult Senegal sole, Solea senegalensis specimens. The stomach was made up of four distinct layers: mucosa, lamina propria-submucosa-, muscularis and serosa. Surface epithelial, glandular and rodlet cells were present in the mucosa. Cells of the columnar epithelium contained a basal nucleus. Numerous mitochondria, granular endoplasmic reticulum and Golgi apparatus consisting of several parallel cisternae and vesicles were observed in the cytoplasm of these cells. The lysosomes were small, round and dense. The gastric glands were numerous in the pyloric and fundic regions but absent in the cardiac stomach. These glands were formed by two cell-types: light and dark cells. The light cells were characterised by numerous mitochondria, while dark cells had slightly fewer mitochondria and a tubulo-vesicular system. Rodlet cells similar to those observed in other teleostean fish were present among the epithelial cells. Although the epithelial cells of the mucosa contained a weak presence of neutral and acid mucopolysaccharides/mucosubstances, these substances were abundant in the lamina propria-submucosa. Proteins rich in arginine, lysine, cysteine and cystine were rarely present in the mucosa and lamina propria-submucosa of stomach, while proteins rich in tyrosine were abundant in these layers. Acid phosphatase, and ATP-ase (pH 7.2 and 9.4) activities were detected in the mucosa and lamina propria-submucosa. Alkaline phosphatase activity was not detected.     

Histological and histochemical observations of the hemolymph cells in the crayfish, Orconectes virilis

Hemolymph cells of Orconectes virilis were stained during the months of August to November by a variety of histological and histochemical techniques. Cells were classified as hyaline cells, small granulocytes, and large granulocytes. Presence of mitochondrial enzymes was indicated by tests for succinic dehydrogenase and cytochrome oxidase. Reaction to test for the hydrolytic enzyme, leucine acylnaphthylamidase was intense in the granules of the large granulocytes. The PAS reaction indicated a mucopolysaccharide at the cell membrane. Lipid was found in all three hemolymph cell types of Orconectes virilis at the time of this study.