Ana Luísa Santos

Mycocerosic acid biomarkers for the diagnosis of tuberculosis in the Coimbra Skeletal Collection

Tuberculosis has been a scourge of humans over many millennia, but questions remain regarding its evolution and epidemiology. Fossil biomarkers, such as DNA and long-chain mycolic acids, can be detected in ancient skeletal and other materials. The phthiocerol dimycocerosate waxes are also robust biomarkers for tuberculosis and sensitive methods are available for the detection of their mycocerosic acid components. The presence of mycocerosic acids was investigated in 49 individuals from the 1837-1936 Coimbra Identified Skeletal Collection (Portugal), half with documentary data indicating tuberculosis as a cause of death. Samples were hydrolysed, acidic components converted to pentafluorobenzyl esters, the non-hydroxylated long-chain esters isolated, and this fraction separated into multimethyl-branched and other esters by normal phase high performance liquid chromatography. Negative ion chemical ionisation gas chromatography mass spectrometry was used to detect diagnostic C29, C30 and C32 mycocerosic acids. Mycocerosic acids were detected in archaeological material for the first time, illustrating that they are valuable biomarkers for the diagnosis of ancient tuberculosis. A 72% correlation with the Coimbra burial record supported TB as the major cause of death. In addition, 30% of the skeletons, positive for mycocerosates, showed the presence of related long-chain mycolipenic acids.

Assessing the influence of occupational and physical activity on the rate of degenerative change of the pubic symphysis in Portuguese males from the 19th to 20th century.

Among the many factors suggested as potential sources of variation in the rate of ageing of the pubic symphysis is occupational/physical activity, but this has yet to be demonstrated. This studys main goal is to determine whether occupational/physical activity influences the aging process in the pubic symphysis. We analyzed a sample of 161 males (18-96 years) from two Portuguese identified skeletal collections, where age-related features were recorded in the pubic symphysis as absent/present. The sample was divided in two groups according to the occupation of the individuals (manual and non-manual) and in two groups according to the femoral robusticity index (robust and gracile). The manual and robust groups represent individuals with more physically demanding occupations while the non-manual and gracile groups represent less physically demanding occupations. The possible influence of occupational/physical activity on the rate of ageing was evaluated by comparing the transition age for each age-related feature, between the two groups, using logistic regression. Only the ligamentous outgrowths on the ventral beveling showed a statistically significant younger age in the robust group, indicating a possible faster rate or early timing of change in the individuals with a more physically demanding activity compared with their gracile counterparts. Thus, these results do not provide significant support to the hypothesis that individuals with a greater demanding occupation or physical activity show greater pubic symphysis degeneration. Further testing of this hypothesis in other samples with individuals from known occupation is required.

The Coimbra Workshop in Musculoskeletal Stress Markers (MSM): an annotated review

The Workshop in Musculoskeletal Stress Markers (MSM): limitations and achievements in the reconstruction of past activity patterns, was held at the University of Coimbra, in Portugal (2 nd-3 rd July 2009). The chief goal of the organizing committee was to provide a productive environment where researchers could discuss the methodologies and terminology employed for the study of MSM as well as assessing how well these enthesopathies serve as markers of occupational stress. The Workshop was organized in three major sessions, each one introduced by a keynote lecture: (1) Methodological issues; (2) MSM studies of archaeological samples and Identified Skeletal Collections; and (3) The multifactorial etiology of enthesopathies. The meeting was concluded with a Plenary Session where two annotators discussed the presentations; this was followed by a general discussion and the formation of three working groups focused on the topics of Methods, Terminology, and Occupations. Details of the Workshop and the presentations presented are available for download at) was created to facilitate communications about individual experiences and to share knowledge among researchers in this field. We hope that this Workshop will help to standardize methodologies and terminology and promote the creation of a more integrated scientific approach to future studies of MSM.

Evidence of hypertrophic osteoarthropathy in individuals from the Coimbra Skeletal Identified Collection (Portugal)

Hypertrophic osteoarthropathy (HOA) is characterized by symmetrical periosteal new bone deposition on the long and short tubular bones. Besides its unknown etiology, it has been associated with pulmonary, cardiovascular and gastrointestinal disorders. The aims of this research were to assess the presence of HOA and to analyze its distribution by sex, age at death, bones affected, type of new bone formed, and cause of death. A sample of 329 individuals (non-adults and adults, of both sexes) was selected from the Coimbra Skeletal Identified Collection. The sample comprised 125 individuals diagnosed with tuberculosis, 64 with pulmonary non-tuberculosis, and 140 with an extrapulmonary and non-tuberculous cause of death. Lesions compatible with HOA were found in all three groups, being statistically significantly in the individuals who died from pulmonary diseases (χ2=5.29, df=1, p=0.02). The risk of developing HOA was 3.41 (χ2=7.77, df=1, p=0.005) higher in the individuals with tuberculosis when compared with the other groups. This work, based on individuals who died before the introduction of antibiotics for treatment, agrees with clinical studies that have established a possible correlation between pulmonary conditions and HOA. These data may help in differentially diagnosing the many instances of periosteal new bone formation found in archaeologically derived skeletal samples.

Bone Diagenesis and its Implication for Disease Diagnosis: The Relevance of Bone Microstructure Analysis for the Study of Past Human Remains.

When bone is exposed to the burial environment it may experience structural changes induced by multiple agents. The study of postmortem alterations is important to differentiate decomposition phenomena from normal physiological processes or pathological lesions, as well as to assess bone tissue quality. Microscopy is of great utility to evaluate the integrity of bone microstructure and it provides significant data on long-term bone decomposition. A total of 18 human bone sections (eight archeological and ten retrieved from an identified skeletal collection) were selected for analysis under plane light and polarized light. The aim of this exploratory study was to analyze the impact of diagenesis and taphonomy on the bone microstructure, as well as on the differential diagnosis of pathological conditions. The results showed that the microscopy approach to bone tissues contributed materially as an aid in the detailed description of the main diagenetic changes observed. It showed that gross inspection does not provide a realistic assessment of bone tissue preservation, which can impact in the characterization of lesions present and subsequent disease diagnosis. Therefore, researchers should continue to consider the application of histological techniques if the aim is to comprehend tissue integrity and its association with decomposition or disease.

Differentiating between rhinosinusitis and mastoiditis surgery from postmortem medical training: A study of two identified skulls and hospital records from early 20th century Coimbra, Portugal

Differentiating between medical procedures performed antemortem, perimortem or postmortem in skeletal remains can be a major challenge. This work aims to present evidence of procedures to treat rhinosinusitis (RS) and mastoiditis, suggest criteria for the diagnosis of frontal sinus disease, and frame the individuals described in their medical historical context. In the International Exchange collection, the skull (878) of a 24-year-old male, who died in 1933 due to frontal sinusitis and meningitis, presents evidence of a trepanation above the right frontonasal suture, and micro/macroporosity on the superciliary arches. The available Coimbra University Hospitals archives (1913-1939) reported that 46 females and 59 males (aged 15 months-84 y.o., x̄=35.33) underwent surgery to treat RS, primarily by trepanation (94.3%). In a search for similar evidence in the collection, the skull of a 42-year-old female (85), who died in 1927 due to sarcoma in the abdomen, shows four quadrangular holes located above the right supraorbital notch, right and left maxilla, and left mastoid process. The number/location of the holes and cut marks point to postmortem medical training (possible dissection). This paper discusses the value of information from historical contexts to differentiate between surgery and medical training in the paleopathological record.

A possible Echinococcus granulosus calcified cyst found in a medieval adult female from the churchyard of Santo Domingo de Silos (Prádena del Rincón, Madrid, Spain)

Calcification, or mineralisation, can occur as part of a natural process, or by pathological processes. The purpose of this work is to examine an unidentified semi-spherical and perforate hollow mass, found near the pelvis of an adult female, dated 12th-13th century AD, exhumed of the Church of Santo Domingo de Silos (Prádena del Ricón, Madrid, Spain). The mass was examined by SEM and Energy Dispersive X-Ray Spectroscopy. These procedures revealed a heterogeneous inner surface with both smooth and irregular areas. A larger spherical and several smaller crescent-shaped perforations were noticed. X-ray microanalysis revealed the presence of the elements C, K, P, Ca, Al, Si, Fe, and Mg. The co-localisation of Ca and P suggests that they may be combined in a mineral matrix, likely formed in vivo. Other minerals probably came from the soil, although Fe could be related to the presence of blood. The macroscopic and microscopic appearances, chemical composition, and location of the calcified mass point to a possible hydatid cyst from Echinococcus granulosus, common in agricultural populations. This study used a suite of analytical techniques that are useful in the diagnosis of unknown calcified masses and can, therefore, be recommended for use in future analytical work.

Archives and Skeletons: an interdisciplinary approach to the study of paleopathology of tuberculosis

Tuberculosis (TB) may be an acute or chronic infection of bone and/or soft tissues. Nevertheless, human skeletons only registered a small percentage of those cases. This work aims to explore data from morbidity and mortality of individuals who contracted or died of TB after Koch bacillus discovery and before the development of antibiotics, using this information to interpret the scarcity of evidences detectable by paleopathological studies. To fulfill these goals, the Coimbra University Hospital and Coimbra Municipality Cemetery records from 1919 to 1928 were analyzed. As expected, pulmonary TB was the most common form of the diseases, both at hospital admission (43.7%, n=904) and as cause of death of the individuals inhumed, especially after the age of 10 years. In children under 10 years, meningeal TB was the more common form. Potts diseases represented only 4.7% (n=98) of the hospital admission due to TB and 0.7% (n=8) of the TB deaths at the cemetery. However, in paleopathology the macroscopic evidence of TB comes mainly from Potts disease. Documented skeletons shows that new bone on visceral surfaces of ribs can occurs in other diseases besides pulmonary TB but its presence can be also use as an indicative of possible TB infection.

Pathological evidence in the Pre-Columbian human remains from the Lee Collection (Jamaica)

The Archaeology Laboratory in the Department of History and Archaeology at UWI has housed the James W. Lee Collection of Arawak artefacts and human remains since 2000. A complete database is being prepared by two of the authors (PAJ and ER) as part of a project under the auspices of the Jamaica Bauxite Institute. The collection spans a total of 265 sites and was collected over a 27 year period (1959-1986). Eight caves and 16 open-air sites produced human remains, with a minimal number of 46 individuals. Despite adverse taphonomic factors, age and sex estimation, metric analyses, and pathological case descriptions, could be carried out in a number of cases. This paper will focus on the paleopathological evidence. Two skulls (EC12, Bull Savannah #2 cave, St. Elizabeth, and CC15, Taylors Hut cave, Clarendon) were artificially modified. CC15 was found inside a bowl. At E12 (Black River West) an increase in thickness in two fragments of tibiae and in one fibula is regarded as a pathological condition, possibly related to treponematosis. Signs of degenerative joint diseases, such as eburnation in an atlas and in a humerus, and osteoarthritis in several bones, were detected at J1 (Hartfield) and JC7 (Spot Valley cave). Finally a case of agenesis exists in the right decidual incisor in a mandible of a juvenile with an age-at-death around 4 years (Y4, Rio Nuevo, St. Mary). This study is a contribution to the understanding of extinct pre-Columbian populations in the Caribbean region.

Skulls and Skeletons from Documented, Overseas and Archaeological Excavations: Portuguese Trajectories

Skulls and skeletons from documented, overseas and archaeological excavations: Portuguese trajectories. In: O’Donnabhain, B.; Lozada, M. (Eds.). Archaeological Human Remains: Legacies of Imperialism, Communism and Colonialism. Cham, Springer, p. 111-125.