First archaeological evidence for ginger consumption as a potential medicinal ingredient in a late medieval leprosarium at St Leonard, Peterborough, England.

Leprosy was one of the most outwardly visible diseases in the European Middle Ages, a period during which leprosaria were founded to provide space for the sick. The extant documentary evidence for leprosy hospitals, especially in relation to diet, therapeutic, and medical care, is limited. However, human dental calculus stands to be an important source of information as it provides insight into the substances people were exposed to and accumulated in their bodies during their lives. In the present study, microremains and DNA were analysed from the calculus of individuals buried in the late medieval cemetery of St Leonard, a leprosarium located in Peterborough, England. The results show the presence of ginger (Zingiber officinale), a culinary and medicinal ingredient, as well as evidence of consumption of cereals and legumes. This research suggests that affected individuals consumed ingredients mentioned in medieval medical textbooks that were used to treat regions of the body typically impacted by leprosy. To the authors' knowledge, this is the first study which has identified Zingiber officinale in human dental calculus in England or on the wider European continent.

Dental anthropological report: Exploring plant-based treatments through the analysis of dental calculus and sediment of dental caries in a woman from the Late Preceramic period, Peru.

Reconstructing plant-based healing treatments of past societies from a dental anthropological perspective is still challenging due to a wide range of plant species, many with both medicinal and nutritional properties, and limitations on plant-taxa identification. Starch grains and phytoliths retrieved in samples from dental calculus and sediment contained in the cavity of dental caries were examined to investigate the supply of a plant-based treatment in an individual buried in the Late Preceramic site of Huaca El Paraíso (2100-1500 BCE), whose osteological analysis reported the absence of any pathological condition at a bone tissue level. A variety of starch grains such as pumpkins, manioc, maize, and beans had an important role in the diet of the individual. The starch grains were embedded in their dental calculus, all of which, except for manioc, showed signs of cooking damage. Considering the context and characteristics of the crops, the nutritional properties of maize, pumpkins and beans are evident. However, parallel medicinal properties of manioc and maize could not be entirely denied. Phytoliths morphologically ascribed to the Asteraceae plant family, which suggest the consumption of medicinal plants of Andean vernacular use, were retrieved in the sediment of dental caries. Our results prove that the analysis of sediment obtained from dental caries is as valuable in tracing medicinal plant-based treatments as dental calculus in archaeological populations. There are still several limitations to approach this topic in dental anthropology, which are discussed in this report. The performance of both analyses -dental calculus and sediment of dental caries, is highly recommended.

The genomic origins of the Bronze Age Tarim Basin mummies.

The identity of the earliest inhabitants of Xinjiang, in the heart of Inner Asia, and the languages that they spoke have long been debated and remain contentious1. Here we present genomic data from 5 individuals dating to around 3000-2800 BC from the Dzungarian Basin and 13 individuals dating to around 2100-1700 BC from the Tarim Basin, representing the earliest yet discovered human remains from North and South Xinjiang, respectively. We find that the Early Bronze Age Dzungarian individuals exhibit a predominantly Afanasievo ancestry with an additional local contribution, and the Early-Middle Bronze Age Tarim individuals contain only a local ancestry. The Tarim individuals from the site of Xiaohe further exhibit strong evidence of milk proteins in their dental calculus, indicating a reliance on dairy pastoralism at the site since its founding. Our results do not support previous hypotheses for the origin of the Tarim mummies, who were argued to be Proto-Tocharian-speaking pastoralists descended from the Afanasievo1,2 or to have originated among the Bactria-Margiana Archaeological Complex3 or Inner Asian Mountain Corridor cultures4. Instead, although Tocharian may have been plausibly introduced to the Dzungarian Basin by Afanasievo migrants during the Early Bronze Age, we find that the earliest Tarim Basin cultures appear to have arisen from a genetically isolated local population that adopted neighbouring pastoralist and agriculturalist practices, which allowed them to settle and thrive along the shifting riverine oases of the Taklamakan Desert.

Dairying enabled Early Bronze Age Yamnaya steppe expansions.

During the Early Bronze Age, populations of the western Eurasian steppe expanded across an immense area of northern Eurasia. Combined archaeological and genetic evidence supports widespread Early Bronze Age population movements out of the Pontic-Caspian steppe that resulted in gene flow across vast distances, linking populations of Yamnaya pastoralists in Scandinavia with pastoral populations (known as the Afanasievo) far to the east in the Altai Mountains1,2 and Mongolia3. Although some models hold that this expansion was the outcome of a newly mobile pastoral economy characterized by horse traction, bulk wagon transport4-6 and regular dietary dependence on meat and milk5, hard evidence for these economic features has not been found. Here we draw on proteomic analysis of dental calculus from individuals from the western Eurasian steppe to demonstrate a major transition in dairying at the start of the Bronze Age. The rapid onset of ubiquitous dairying at a point in time when steppe populations are known to have begun dispersing offers critical insight into a key catalyst of steppe mobility. The identification of horse milk proteins also indicates horse domestication by the Early Bronze Age, which provides support for its role in steppe dispersals. Our results point to a potential epicentre for horse domestication in the Pontic-Caspian steppe by the third millennium BC, and offer strong support for the notion that the novel exploitation of secondary animal products was a key driver of the expansions of Eurasian steppe pastoralists by the Early Bronze Age.

Tracking the transition to agriculture in Southern Europe through ancient DNA analysis of dental calculus.

Archaeological dental calculus, or mineralized plaque, is a key tool to track the evolution of oral microbiota across time in response to processes that impacted our culture and biology, such as the rise of farming during the Neolithic. However, the extent to which the human oral flora changed from prehistory until present has remained elusive due to the scarcity of data on the microbiomes of prehistoric humans. Here, we present our reconstruction of oral microbiomes via shotgun metagenomics of dental calculus in 44 ancient foragers and farmers from two regions playing a pivotal role in the spread of farming across Europe-the Balkans and the Italian Peninsula. We show that the introduction of farming in Southern Europe did not alter significantly the oral microbiomes of local forager groups, and it was in particular associated with a higher abundance of the species Olsenella sp. oral taxon 807. The human oral environment in prehistory was dominated by a microbial species, Anaerolineaceae bacterium oral taxon 439, that diversified geographically. A Near Eastern lineage of this bacterial commensal dispersed with Neolithic farmers and replaced the variant present in the local foragers. Our findings also illustrate that major taxonomic shifts in human oral microbiome composition occurred after the Neolithic and that the functional profile of modern humans evolved in recent times to develop peculiar mechanisms of antibiotic resistance that were previously absent.

Environmental implications and evidence of natural products from dental calculi of a Neolithic-Chalcolithic community (central Italy).

In this contribution, we investigated the role of plants in the prehistoric community of Casale del Dolce (Anagni, FR, central Italy), through microparticles recovered from dental calculus. The finding of a great amount of pollen types, even in form of compact lumps, could indicate use of natural substances, such as honeybee products and/or conifer resins. This plant-microremain record also suggested environmental implications relative to the Neolithic and Chalcolithic period. Additionally, the stability of the tartar microenvironment had preserved starches and other microparticles, such as one epidermal trichome, a sporangium, and fragments of plant tissue, rarely detected in ancient dental calculus. The detection of secondary metabolites in the ancient matrix confirmed the familiarity of this community with plant resources. All these data supply various interesting food for thought and expand the knowledge about the potential of dental calculus in archaeological and archaeobotanical fields with a special focus on palaeoecology.

Exploring late Paleolithic and Mesolithic diet in the Eastern Alpine region of Italy through multiple proxies.

OBJECTIVES: The analysis of prehistoric human dietary habits is key for understanding the effects of paleoenvironmental changes on the evolution of cultural and social human behaviors. In this study, we compare results from zooarchaeological, stable isotope and dental calculus analyses as well as lower second molar macrowear patterns to gain a broader understanding of the diet of three individuals who lived between the end of the Late Pleistocene and the Early Holocene (ca., 17-8 ky cal BP) in the Eastern Alpine region of Italy. MATERIALS AND METHODS: We analyze individuals buried at the sites of Riparo Tagliente (Verona), Riparo Villabruna, and Mondeval de Sora (Belluno). The three burials provide a unique dataset for diachronically exploring the influence of climatic changes on human subsistence strategies. RESULTS: Isotopic results indicate that all individuals likely relied on both terrestrial and freshwater animal proteins. Even though dental calculus analysis was, in part, hindered by the amount of mineral deposit available on the teeth, tooth macrowear study suggests that the dietary habits of the individuals included plant foods. Moreover, differences in macrowear patterns of lower second molars have been documented between Neanderthals and modern humans in the present sample, due to a prevalence of Buccal wear among the former as opposed to higher values of Lingual wear in modern human teeth. DISCUSSION: Isotopic analyses have emphasized the contribution of animal proteins in the diet of the three foragers from the Eastern Alpine region. The possible intake of carbohydrate-rich plant foods, suggested by the retrieval of plant remains in dental calculus, is supported by the signal of macrowear analysis. Moreover, the latter method indicates that the distribution of macrowear in lower second molars (M2 s) allows us to discriminate between Neanderthals and modern humans within the present reference sample. Overall, our results show these three prehistoric hunter-gatherers were well adapted to the environment in which they lived exploiting many natural resources.

Functional diversity of microbial ecologies estimated from ancient human coprolites and dental calculus.

Human microbiome studies are increasingly incorporating macroecological approaches, such as community assembly, network analysis and functional redundancy to more fully characterize the microbiome. Such analyses have not been applied to ancient human microbiomes, preventing insights into human microbiome evolution. We address this issue by analysing published ancient microbiome datasets: coprolites from Rio Zape (n = 7; 700 CE Mexico) and historic dental calculus (n = 44; 1770-1855 CE, UK), as well as two novel dental calculus datasets: Maya (n = 7; 170 BCE-885 CE, Belize) and Nuragic Sardinians (n = 11; 1400-850 BCE, Italy). Periodontitis-associated bacteria (Treponema denticola, Fusobacterium nucleatum and Eubacterium saphenum) were identified as keystone taxa in the dental calculus datasets. Coprolite keystone taxa included known short-chain fatty acid producers (Eubacterium biforme, Phascolarctobacterium succinatutens) and potentially disease-associated bacteria (Escherichia, Brachyspira). Overlap in ecological profiles between ancient and modern microbiomes was indicated by similarity in functional response diversity profiles between contemporary hunter-gatherers and ancient coprolites, as well as parallels between ancient Maya, historic UK, and modern Spanish dental calculus; however, the ancient Nuragic dental calculus shows a distinct ecological structure. We detected key ecological signatures from ancient microbiome data, paving the way to expand understanding of human microbiome evolution. This article is part of the theme issue 'Insights into health and disease from ancient biomolecules'.

New insights on Neolithic food and mobility patterns in Mediterranean coastal populations.

OBJECTIVES: The aims of this research are to explore the diet, mobility, social organization, and environmental exploitation patterns of early Mediterranean farmers, particularly the role of marine and plant resources in these foodways. In addition, this work strives to document possible gendered patterns of behavior linked to the neolithization of this ecologically rich area. To achieve this, a set of multiproxy analyses (isotopic analyses, dental calculus, microremains analysis, ancient DNA) were performed on an exceptional deposit (n = 61) of human remains from the Les Bréguières site (France), dating to the transition of the sixth to the fifth millennium BCE. MATERIALS AND METHODS: The samples used in this study were excavated from the Les Bréguières site (Mougins, Alpes-Maritimes, France), located along the southeastern Mediterranean coastline of France. Stable isotope analyses (C, N) on bone collagen (17 coxal bones, 35 craniofacial elements) were performed as a means to infer protein intake during tissue development. Sulfur isotope ratios were used as indicators of geographical and environmental points of origin. The study of ancient dental calculus helped document the consumption of plants. Strontium isotope analysis on tooth enamel (n = 56) was conducted to infer human provenance and territorial mobility. Finally, ancient DNA analysis was performed to study maternal versus paternal diversity within this Neolithic group (n = 30). RESULTS: Stable isotope ratios for human bones range from -20.3 to -18.1‰ for C, from 8.9 to 11.1‰ for N and from 6.4 to 15‰ for S. Domestic animal data range from -22.0 to -20.2‰ for C, from 4.1 to 6.9‰ for N, and from 10.2 to 12.5‰ for S. Human enamel 87 Sr/86 Sr range from 0.7081 to 0.7102, slightly wider than the animal range (between 0.7087 and 0.7096). Starch and phytolith microremains were recovered as well as other types of remains (e.g., hairs, diatoms, fungal spores). Starch grains include Triticeae type and phytolith includes dicotyledons and monocot types as panicoid grasses. Mitochondrial DNA characterized eight different maternal lineages: H1, H3, HV (5.26%), J (10.53%), J1, K, T (5.2%), and U5 (10.53%) but no sample yielded reproducible Y chromosome SNPs, preventing paternal lineage characterization. DISCUSSION: Carbon and nitrogen stable isotope ratios indicate a consumption of protein by humans mainly focused on terrestrial animals and possible exploitation of marine resources for one male and one undetermined adult. Sulfur stable isotope ratios allowed distinguishing groups with different geographical origins, including two females possibly more exposed to the sea spray effect. While strontium isotope data do not indicate different origins for the individuals, mitochondrial lineage diversity from petrous bone DNA suggests the burial includes genetically differentiated groups or a group practicing patrilocality. Moreover, the diversity of plant microremains recorded in dental calculus provide the first evidence that the groups of Les Bréguières consumed a wide breadth of plant foods (as cereals and wild taxa) that required access to diverse environments. This transdisciplinary research paves the way for new perspectives and highlights the relevance for novel research of contexts (whether recently discovered or in museum collections) excavated near shorelines, due to the richness of the biodiversity and the wide range of edible resources available.

Exploitation and utilization of tropical rainforests indicated in dental calculus of ancient Oceanic Lapita culture colonists.

Remote Oceania, which largely consists of islands covered in tropical forests, was the last region on earth to be successfully colonized by humans, beginning 3,000 years ago. We examined human dental calculus from burials in an ancient Lapita culture cemetery to gain insight into the early settlement of this previously untouched tropical environment, specifically on the island of Efate in Vanuatu. Dental calculus is an ideal material to analyse questions of human and plant interactions due to the ingestion of plant-derived microparticles that become incorporated into the calculus as it forms throughout a person's life. Most of the microparticles identified here are from tree and shrub resources, including a ~2,900 calibrated (cal) BP example of banana in Remote Oceania, providing direct evidence for the importance of forests and arboriculture during the settlement of Remote Oceania.

Manifestações bucais e o perfil bioquímico salivar de mulheres com anorexia e bulimia nervosa / Oral manifestations and the salivary biochemical profile of women with anorexia and bulimia nervosa

Introdução: Os distúrbios alimentares são transtornos psiquiátricos graves, com altos níveis de mortalidade, incapacidade e morbidade física e psicológica; o que provoca redução na qualidade de vida. Objetivo: O presente trabalho teve como objetivos realizar uma revisão sistemática com base na análise de publicações nacionais e internacionais presentes na literatura científica sobre a ocorrência de manifestações bucais em pacientes com anorexia e bulimia nervosa; comparar a ocorrência de cárie, erosão dentária e o perfil bioquímico salivar de mulheres com e sem transtornos alimentares; além de relatar a condição de saúde bucal de uma paciente com anorexia nervosa internada em um hospital de psiquiatria pertencente à Faculdade de Medicina de Marília (FAMEMA), São Paulo, Brasil. Metodologia: A tese foi dividida em três partes. No primeiro momento da pesquisa, realizou-se vasto levantamento bibliográfico para a elaboração do primeiro capítulo, que consistiu em uma revisão sistemática sobre o assunto. O segundo capítulo foi composto por um estudo observacional do tipo casocontrole. Realizou-se a coleta de dados com os pacientes diagnosticados com bulimia e anorexia nervosa e que estavam sob tratamento no ambulatório. O grupo controle foi composto por acompanhantes dos doentes, que possuíam características similares. O terceiro capítulo foi composto por um caso clínico de uma paciente que estava hospitalizada com anorexia nervosa no Hospital das Clínicas da FAMEMA (Unidade I), onde foi realizada toda a descrição de suas condições de saúde bucal. Resultados: No capítulo 1, foi obtido um número final de seis trabalhos, onde todos respondiam a questão sobre quais manifestações bucais são encontradas em pacientes com anorexia e bulimia nervosa, e apresentavam validade e poder em sua estrutura metodológica e resultados. No segundo capítulo, podemos observar que, o grupo das pacientes com anorexia e bulimia nervosa apresenta maiores índices de cárie e erosão dentária, assim como alterações nos parâmetros salivares, em especial o fluxo salivar e os marcadores bioquímicos fósforo, cálcio, FRAP e ácido úrico; quando comparados ao grupo controle. E por fim, na terceira parte da presente tese, observamos que a paciente apresentava 11 dentes atacados pela cárie dentária. No exame periodontal, todos os dentes sondados apresentaram sangramento, além de cálculo nos incisivos inferiores e molares superiores pela face lingual; e o fluxo salivar da paciente encontrava-se extremamente reduzido. Quanto aos parâmetros salivares, o valor encontrado para o cálcio e fósforo estavam aumentados, 13,36mg/dL e 6,31mg/dL, respectivamente. Conclusão: Dessa forma, conclui-se que pacientes diagnosticados com anorexia e bulimia nervosa normalmente apresentam lesões cariosas e erosivas nos dentes e gengivas, além de alterações nos parâmetros bioquímicos salivares(AU)

Introduction: Eating disorders are serious psychiatric disorders, with high levels of mortality, disability and physical and psychological morbidity; which causes a reduction in quality of life. Objective: The present study aimed to carry out a systematic review based on the analysis of national and international publications present in the scientific literature on the occurrence of oral manifestations in patients with anorexia and bulimia nervosa; to compare the occurrence of caries, dental erosion and the biochemical salivary profile of women with and without eating disorders; in addition to reporting the oral health condition of a patient with anorexia nervosa admitted to a psychiatric hospital belonging to the Faculty of Medicine of Marília (FAMEMA), São Paulo, Brazil. Methodology: The thesis was divided into three parts. In the first moment of the research, a vast bibliographic survey was carried out for the elaboration of the first chapter, which consisted of a systematic review on the subject. The second chapter consisted of an observational case-control study. Data collection was performed with patients diagnosed with bulimia and anorexia nervosa and who were under treatment at the outpatient clinic. The control group was composed of patients' companions, who had similar characteristics. The third chapter consisted of a clinical case of a patient who was hospitalized with anorexia nervosa at Hospital das Clínicas da FAMEMA (Unit I), where the entire description of her oral health conditions was carried out. Results: In chapter 1, a final number of six studies was obtained, in which all answered the question about which oral manifestations are found in patients with anorexia and bulimia nervosa, and presented validity and power in their methodological structure and results. In the second chapter, we can see that the group of patients with anorexia and bulimia nervosa has higher rates of caries and dental erosion, as well as changes in salivary parameters, especially the salivary flow and the biochemical markers phosphorus, calcium, FRAP and uric acid ; when compared to the control group. Finally, in the third part of this thesis, we observed that the patient had 11 teeth attacked by dental caries. In the periodontal examination, all probed teeth showed bleeding, in addition to calculus in the lower incisors and upper molars through the lingual surface; and the patient's salivary flow was extremely reduced. As for salivary parameters, the values found for calcium and phosphorus were increased, 13.36mg/dL and 6.31mg/dL, respectively. Conclusion: Thus, it is concluded that patients diagnosed with anorexia and bulimia nervosa usually have carious and erosive lesions on the teeth and gums, in addition to changes in the salivary biochemical parameters(AU)

Lifestyle of a Roman Imperial community: ethnobotanical evidence from dental calculus of the Ager Curensis inhabitants.

BACKGROUND: The analysis of ancient calcified dental plaque is a powerful archaeobotanical method to elucidate the key role of the plants in human history. METHODS: In this research, by applying both optic microscopy and gas chromatography mass spectrometry on this matrix, a detailed qualitative investigation for reconstructing the lifestyle of a Roman imperial community of the Ager Curensis (Sabina Tiberina, Central Italy) was performed. RESULTS: The detection of animal micro-remains and molecules (e.g., hairs, feather barbules, markers of dairy products), starch granules of several cereals and legumes, pollen (e.g., Juglans regia L., Hedera sp. L.) and other plant micro-debris (e.g., trichome of Olea sp., hemp fibers), and phytochemicals (e.g., Brassicaceae, Lamiaceae herbs, Ferula sp., Trigonella foenum-graecum L., wine, and Humulus lupulus L.) in the dental calculus sample demonstrated that plant-derived foods were regularly consumed together with animal resources. CONCLUSIONS: This nutritional plan, consistent with the information reported in ancient written texts, suggested that the studied population based its own subsistence on both agriculture and husbandry, probably also including beekeeping and hunting activities. All together, these results represent proofs for the comprehension of food habits, phytotherapeutic practices, and cultural traditions of one of the first Roman settlements in the Sabina Tiberina area.

Two Randomized Trials Demonstrate Lactic Acid Supplementation in Pet Food Inhibits Dental Plaque, Calculus, and Tooth Stain in Cats.

BACKGROUND: Dental concerns are some of the most common health problems affecting companion animals. A variety of foods, treats, and chews comprising different mechanical and chemical technologies have been investigated as a means of promoting oral health. Here, we investigate the chemical technology, lactic acid added to a commercially available food, for its ability to inhibit dental plaque, calculus, and tooth stain accumulation in cats. METHODS: Two separate feeding trials assessed the utility of a nutritionally complete feline maintenance food supplemented with lactic acid to reduce oral substrate accumulation (dental plaque, calculus, and tooth stain) in cats. After a calibration study identified high and low dental plaque formers, 45 cats were randomized to 1 of 2 test groups (food with 1.2% lactic acid supplementation) or control (food without lactic acid supplementation) groups, stratified based on their calibration scores. Data were collected on a monthly basis for 3 months. The second study randomly assigned 24 cats to either the test or control groups for 1 year, with data collected at the 6- and 12-month time points. RESULTS: In the 3-month study, reductions in dental plaque, calculus, and tooth stain accumulations were observed at the 2-month assessment in both test groups compared with control (P < .05 for test group 2). The 1-year study showed that these reductions in oral substrate accumulation persisted through the 6- and 12-month time points (P < .05). CONCLUSIONS: Taken together, these studies demonstrate that lactic acid supplemented at 1.2% in a feline maintenance food significantly inhibits oral substrate accumulation.

Tooth grooves, occlusal striations, dental calculus, and evidence for fiber processing in an Italian eneolithic/bronze age cemetery.

OBJECTIVES: We conducted a systematic macroscopic and microscopic examination of occlusal and para-occlusal wear in a large dental sample (n = 3,014) from 217 individuals dated to the Early Bronze age site of Gricignano d'Aversa, Italy. We used macroscopic and microscopic techniques to document nondietary occlusal and para-occlusal wear and to analyze calculus inclusions in some of the teeth. In combining an analysis of the wear with the calculus inclusions we linked the specific wear to the likely fiber that was involved in producing it. MATERIALS AND METHODS: Teeth and their high resolution epoxy casts were analyzed through SEM and reflected light microscopes. Nineteen individuals (fifteen with activity induced dental modifications and four as a control sample) were examined for the presence of calculus inclusions. RESULTS: Activity induced dental modifications (AIDMs), notches, grooves and micro-striations, were found in the 62.2% of the adult females, in 21.2% of the adults of unknown sex and in a single male. We found the full spectrum of dental manipulations from very minor nonocclusal wear in some young individuals to severe attrition at the other extreme. The width of the striations and grooves, mostly on the upper incisors, suggests a craft activity involving fibers and thread production and manipulation. From the dental calculus of two females with grooves and striations, we extracted three fragments of fibers, identified as hemp (Cannabis, sp.). Previously from Gricignano woven hemp fibers were found on both surfaces of a metal blade associated with a male burial. DISCUSSION: This study found the co-occurrence of tooth AIDMs and the actual fibers preserved in the dental calculus. As more work is done analyzing dental calculus in a variety of humans, it is apparent that this biological material holds rich resources documenting non-dietary habits.

Proteomic evidence of dietary sources in ancient dental calculus.

Archaeological dental calculus has emerged as a rich source of ancient biomolecules, including proteins. Previous analyses of proteins extracted from ancient dental calculus revealed the presence of the dietary milk protein ß-lactoglobulin, providing direct evidence of dairy consumption in the archaeological record. However, the potential for calculus to preserve other food-related proteins has not yet been systematically explored. Here we analyse shotgun metaproteomic data from 100 archaeological dental calculus samples ranging from the Iron Age to the post-medieval period (eighth century BC to nineteenth century AD) in England, as well as 14 dental calculus samples from contemporary dental patients and recently deceased individuals, to characterize the range and extent of dietary proteins preserved in dental calculus. In addition to milk proteins, we detect proteomic evidence of foodstuffs such as cereals and plant products, as well as the digestive enzyme salivary amylase. We discuss the importance of optimized protein extraction methods, data analysis approaches and authentication strategies in the identification of dietary proteins from archaeological dental calculus. This study demonstrates that proteomic approaches can robustly identify foodstuffs in the archaeological record that are typically under-represented due to their poor macroscopic preservation.

A grazing Gomphotherium in Middle Miocene Central Asia, 10 million years prior to the origin of the Elephantidae.

Feeding preference of fossil herbivorous mammals, concerning the coevolution of mammalian and floral ecosystems, has become of key research interest. In this paper, phytoliths in dental calculus from two gomphotheriid proboscideans of the middle Miocene Junggar Basin, Central Asia, have been identified, suggesting that Gomphotherium connexum was a mixed feeder, while the phytoliths from G. steinheimense indicates grazing preference. This is the earliest-known proboscidean with a predominantly grazing habit. These results are further confirmed by microwear and isotope analyses. Pollen record reveals an open steppic environment with few trees, indicating an early aridity phase in the Asian interior during the Mid-Miocene Climate Optimum, which might urge a diet remodeling of G. steinheimense. Morphological and cladistic analyses show that G. steinheimense comprises the sister taxon of tetralophodont gomphotheres, which were believed to be the general ancestral stock of derived "true elephantids"; whereas G. connexum represents a more conservative lineage in both feeding behavior and tooth morphology, which subsequently became completely extinct. Therefore, grazing by G. steinheimense may have acted as a behavior preadaptive for aridity, and allowing its lineage evolving new morphological features for surviving later in time. This study displays an interesting example of behavioral adaptation prior to morphological modification.

Efficacy of a Kalanchoe gastonis-bonnieri extract to control bacterial biofilms and dental calculus in dogs / Eficácia do extrato de Kalanchoe gastonis-bonnieri no controle do biofilme bacteriano e cálculo dentário em cães

An aqueous leaf extract of the medicinal species Kalanchoe gastonis-bonnieri (here denominated KGB) has been found to be effective as an antimicrobial agent against canine oral cavity bacteria in in vitro assays. In this study, we investigated the effect of topic oral administration of KGB on the development of dental biofilm in Beagle dogs. The experiments were performed with an experimental group (0.2% of KGB extract), a negative control group (0.9% of saline solution) and a positive control group (0.12% chlorhexidine). Each treatment was sprayed into the oral cavity daily for 28 days. Thirty Beagle dogs with similar characteristics and kept under the same management and diet were used. The measurement of dental plaque and calculus was performed using a computerized analytical method. The phenolic profile of KGB extract was analyzed by HPLC-DAD. KGB extract at 0.2% showed efficacy in controlling the formation of plaque compared to the negative control group, and dental calculus in relation to the negative and positive control groups. A significant difference was observed among these three groups. Peaks attributed to flavonoids and phenolic acids were identified in the HPLC-DAD chromatogram of the KGB extract. The presence of these substances could be related to the activity observed. Our findings demonstrate that treatment with KGB is effective in controlling periodontal disease in dogs, providing new insights into the medicinal properties of this plant. KGB extract has a potential use as a supplemental agent in pharmaceutical products for the prevention of periodontal disease.(AU)

Um extrato aquoso de folhas da espécie medicinal Kalanchoe gastonis-bonnieri (aqui denominado como KGB) foi efetivo como um agente antimicrobiano contra as bactérias da cavidade oral de cães em testes in vitro. Neste estudo, investigou-se o efeito da administração oral tópica de KGB sobre o desenvolvimento do biofilme dental em cães da raça Beagle. Os experimentos foram realizados com um grupo experimental (0,2% de extrato de KGB), um grupo controle negativo (0,9% de solução salina) e um grupo controle positivo (0,12% de gluconato de clorexidina). Cada tratamento foi aplicado no interior da cavidade oral diariamente durante 28 dias. Foram utilizados trinta cães da raça Beagle com características semelhantes e mantidos sob o mesmo manejo e dieta. A medição da placa bacteriana e cálculo dentários foi realizada utilizando-se um método de análise computadorizada. O perfil fenólico do extrato de KGB foi analisado por HPLC-DAD. O extrato de KGB a 0,2% mostrou eficácia no controle da formação de placa bacteriana em comparação com o grupo controle negativo, e de cálculo dentário em relação aos grupos controle negativo e positivo. Uma diferença significativa foi observada entre esses três grupos. Picos atribuídos a flavonoides e ácidos fenólicos foram identificados no cromatograma de HPLC-DAD do extrato de KGB. A presença de tais substâncias pode estar relacionada com a atividade observada. Os resultados demonstram que o tratamento com KGB é eficaz no controle da doença periodontal em cães, fornecendo novas perspectivas sobre as propriedades medicinais desta planta. O extrato de KGB tem uma utilização potencial como um agente suplementar em produtos farmacêuticos para a prevenção da doença periodontal.(AU)

Neanderthal behaviour, diet, and disease inferred from ancient DNA in dental calculus.

Recent genomic data have revealed multiple interactions between Neanderthals and modern humans, but there is currently little genetic evidence regarding Neanderthal behaviour, diet, or disease. Here we describe the shotgun-sequencing of ancient DNA from five specimens of Neanderthal calcified dental plaque (calculus) and the characterization of regional differences in Neanderthal ecology. At Spy cave, Belgium, Neanderthal diet was heavily meat based and included woolly rhinoceros and wild sheep (mouflon), characteristic of a steppe environment. In contrast, no meat was detected in the diet of Neanderthals from El Sidrón cave, Spain, and dietary components of mushrooms, pine nuts, and moss reflected forest gathering. Differences in diet were also linked to an overall shift in the oral bacterial community (microbiota) and suggested that meat consumption contributed to substantial variation within Neanderthal microbiota. Evidence for self-medication was detected in an El Sidrón Neanderthal with a dental abscess and a chronic gastrointestinal pathogen (Enterocytozoon bieneusi). Metagenomic data from this individual also contained a nearly complete genome of the archaeal commensal Methanobrevibacter oralis (10.2× depth of coverage)-the oldest draft microbial genome generated to date, at around 48,000 years old. DNA preserved within dental calculus represents a notable source of information about the behaviour and health of ancient hominin specimens, as well as a unique system that is useful for the study of long-term microbial evolution.

Dental calculus reveals Mesolithic foragers in the Balkans consumed domesticated plant foods.

Researchers agree that domesticated plants were introduced into southeast Europe from southwest Asia as a part of a Neolithic "package," which included domesticated animals and artifacts typical of farming communities. It is commonly believed that this package reached inland areas of the Balkans by ∼6200 calibrated (cal.) BC or later. Our analysis of the starch record entrapped in dental calculus of Mesolithic human teeth at the site of Vlasac in the Danube Gorges of the central Balkans provides direct evidence that already by ∼6600 cal. BC, if not earlier, Late Mesolithic foragers of this region consumed domestic cereals, such as Triticum monococcum, Triticum dicoccum, and Hordeum distichon, which were also the main crops found among Early Neolithic communities of southeast Europe. We infer that "exotic" Neolithic domesticated plants were introduced to southern Europe independently almost half a millennium earlier than previously thought, through networks that enabled exchanges between inland Mesolithic foragers and early farming groups found along the Aegean coast of Turkey.

Mineral density volume gradients in normal and diseased human tissues.

Clinical computed tomography provides a single mineral density (MD) value for heterogeneous calcified tissues containing early and late stage pathologic formations. The novel aspect of this study is that, it extends current quantitative methods of mapping mineral density gradients to three dimensions, discretizes early and late mineralized stages, identifies elemental distribution in discretized volumes, and correlates measured MD with respective calcium (Ca) to phosphorus (P) and Ca to zinc (Zn) elemental ratios. To accomplish this, MD variations identified using polychromatic radiation from a high resolution micro-computed tomography (micro-CT) benchtop unit were correlated with elemental mapping obtained from a microprobe X-ray fluorescence (XRF) using synchrotron monochromatic radiation. Digital segmentation of tomograms from normal and diseased tissues (N=5 per group; 40-60 year old males) contained significant mineral density variations (enamel: 2820-3095 mg/cc, bone: 570-1415 mg/cc, cementum: 1240-1340 mg/cc, dentin: 1480-1590 mg/cc, cementum affected by periodontitis: 1100-1220 mg/cc, hypomineralized carious dentin: 345-1450 mg/cc, hypermineralized carious dentin: 1815-2740 mg/cc, and dental calculus: 1290-1770 mg/cc). A plausible linear correlation between segmented MD volumes and elemental ratios within these volumes was established, and Ca/P ratios for dentin (1.49), hypomineralized dentin (0.32-0.46), cementum (1.51), and bone (1.68) were observed. Furthermore, varying Ca/Zn ratios were distinguished in adapted compared to normal tissues, such as in bone (855-2765) and in cementum (595-990), highlighting Zn as an influential element in prompting observed adaptive properties. Hence, results provide insights on mineral density gradients with elemental concentrations and elemental footprints that in turn could aid in elucidating mechanistic processes for pathologic formations.