Carbon and Nitrogen stable isotope analysis of bone collagen is at present an almost routine method in the study of ancient diets. By stable isotope analysis it is possible to determine directly the type of diet of an individual and consequently of a population group. Therefore, at regional levels, the similarities and differences in the diet of individuals within an entire population can be easily detected (Katzenberg, 2000). This is a preliminary report of a more complex study, still in course, of 25 individuals from the Basilica of S. Domenico Maggiore in Naples (15th-17th centuries) (Fornaciari, 1999, 2006) and of 20 individuals from the Medici Chapels of the Basilica of S. Lorenzo in Florence (16th-17th centuries) (Fornaciari, et al, 2007). The Italian Renaissance princes had a much wider choice of food than other classes (Grieco, 1999). For this reason their diet should not be considered typical of the wealthy class as a whole, in the same way as the isotopic data observed in the population groups sampled for this study may not be representative for all the groups of the elite Italian society. Historical records reveal that after meat and wine, which constituted the central nucleus of the nobles’ diet, eggs and cheese also appear, although much less frequently (Riera Melis, 1999). Th ese two types of food, with high protein content, were combined with meat on normal days and alternated with fish on penitential occasions. In the aristocratic dietary regimen, vegetables occupied a secondary place, with almost total absence of fruit (Grieco, 1999). The isotope values obtained are remarkably similar to those published for other late Medieval and post-Medieval populations in England and clearly refl ect the large intake not only of meat but also of marine foods by the Italian aristocratic classes, especially from southern Italy, in the 15th-17th centuries (Fornaciari, 2008). Other foods high in proteins and enriched in nitrogen, such as freshwater fi sh, birds (and their eggs), or mollusks are likely to have provided smaller contributions. Given the large variety of dietary resources attested by historical documents, this conclusion can hardly be surprising. I present three “clinical” cases, important from paleonutritional and paleopathological point of view. Th e left foot of Ferdinando I de’ Medici, Grand Duke of Tuscany (1549-1609), shows a scoped-out defect located at the peri-articular and articular surface of the interphalangeal joint of the hallux dorsum, with partial destruction of the sub-chondral plate; the lesion, revealing at X-ray an evident sclerotic margin, involves both the bones of the joint and is typical of chronic gout (Fornaciari et al, 2007). High values of δ15N demonstrate a diet very rich in meat from terrestrial animals. δ13C values show a minor integration with foods of marine origin (fi sh) (Fornaciari, 2008). This isotopic profile well correlates with the frequent attacks of gout referred by court chroniclers and with the diagnosis of chronic gout of the left big toe revealed by the paleopathological study. The skull of Don Filippino de’ Medici (1577-1582), eldest son of the Grand Duke Francesco I, reveals non-severe external hydrocephaly, as witnessed by the macroscopic and radiological study, and by a 16th century autopsy report. Rickets also worsened this pathological condition (Fornaciari et al., 2007). The δ15N isotope value of don Filippino is anomalous and intermediate between the very high levels of breast-feeding babies and those of post-weaning children. This value reveals a diet very rich in proteins of animal origin. If we plot the δ15N and δ13C values together, we can see that don Filippino shows a very anomalous and isolated position. Isotopic signals reveal a strong intake of terrestrial animal proteins, while the level of marine proteins falls within the normal range of the group (Fornaciari, 2008). We are in front of a really unbalanced diet of the child, with regard to either meat or milk. On the basis of the historical, paleonutritional and paleopathological data we can speculate about the life of don Filippino. The eldest son of the Grand Duke, the hereditary prince, was a frail and sickly child, with frequent indispositions and persistent fevers, aff ected by rickets and a light form of hydrocephaly. Probably for this reason, his parents and the court doctors worried about the child’s health, and forced him to eat more meat, considered at that time “the fi rst source of physical strength”(Riera-Melis, 1999). Th is fact would explain the presence of this isotopic signal in the bones of the small prince. Autopsy of the mummy of Ferrante I d’Aragona, king of Naples (1431-1494), revealed a moderately differentiated colon adenocarcinoma extensively infi ltrating the muscles of the small pelvis. Histology showed exceptionally well preserved neoplastic cells, disposed in cords and solid nests or forming pseudo-glandular lumina, disseminated in a fibrous tissue containing striated muscular fi bres. Th e cells appeared tall and crowded, with abundant cytoplasm and quite regular hyperchromatic nuclei (Fornaciari et al, 1993). The excellent, almost exceptional preservation of the histological structure of this unique tumor is probably due to the very dry microclimatic conditions of the church and certainly to the preservation substances used for embalming. Ancient DNA amplification of the neoplastic cells by polymerase chain reaction (PCR) in the mummy of Ferrante I evidenced a typical point mutation of the K-ras gene codon 12 (Marchetti, et al, 1996; Falchetti, et al. 2006). The recorded genetic change represents the most frequent mutation of the K-ras gene in sporadic colorectal cancer and is characteristic of the eff ects of a number of “alkylating” agents. Th e N-nitroso compounds (NOC) and their precursors, have been widely investigated for their diff usion and well known relevance in gastro-intestinal cancerogenesis. In particular the carcinogenic properties of N-nitroso compounds (NOC) are also associated with their ability to “alkylate” DNA. Recent studies have focused on the importance of endogenous NOC (Soussi, et al, 2007). The diet contains many constituents which can be nitrosated in the gastrointestinal tract (especially in the stomach). Increased intake of red meat induces a significant (3-fold) increase of faecal N-nitroso compound (NOC) levels, with a range of exposure in the faeces similar to that from tobacco-specific NOC in cigarette smokers. Th is endogenous formation of NOC in the human colon well explains the association between red meat and colorectal cancer (Fornaciari, et al, 1999). The portraits of Ferrante, both sculptures and pictures, reveal growing obesity from youth to maturity. Court chroniclers report on the strong preference of the king for red meat, especially game. Examination of the mummy of Ferrante, even if severely damaged by a fi re in 1509, confirms his obesity. Histology reveals severe arteriosclerosis of the carotid arteries. Finally, the paleonutritional data, with their high level of δ15N, show a massive intake of animal proteins (meat). The alimentary “environment” of the Neapolitan court of the XV century and the sovereign’s habits, with his abundance of natural endogenous alkylating agents, can well explain the K-ras mutation causing the tumor which killed the Aragonese king over fi ve centuries ago (Fornaciari, et al, 1999). In conclusion, the investigation into the diet of the Renaissance aristocratic classes in Italy well demonstrates the potential of the studies on Carbon and Nitrogen stable isotope analysis in archeology and paleopathology. REFERENCES 1. Falchetti M, Lupi R, Ottini L. Molecular analysis of a colorectal carcinoma from a mummy of the XVth century. Journal of History of Medicine. 2006;18:943-951. 2. Fornaciari G, Castagna M, Naccarato AG, Tognetti A, Collecchi P, Bevilacqua G. Adenocarcinoma in the mummy of Ferrante I of Aragon, King of Naples (1431-1494). Paleopathology Newsletter. 1993;82:7-11. 3. Fornaciari G, Marchetti A, Pellegrini S, Ciranni R. K-ras mutation in the tumour of King Ferrante I of Aragon (1431-1494) and environmental mutagens at the Aragonese court of Naples. International Journal of Osteoarcheology. 1999;9:297-304. 4. Fornaciari G. Renaissance mummies in Italy. Medicina nei Secoli. 1999;11:85-105. 5. Fornaciari G. Le mummie aragonesi in San Domenico Maggiore di Napoli. Medicina nei Secoli. 2006;18:875-896. 6. Fornaciari G, Vitiello A, Giusiani S, Giuffra V, Fornaciari A, Villari N. The Medici Project: fi rst anthropological and paleopathological results of the exploration of the Medici tombs in Florence. Medicina nei Secoli. 2007;19:521-544. 7. Fornaciari G. Food and disease at the Renaissance courts of Naples and Florence: a paleonutritional study. Appetite. 2008. In press. 8. Grieco AJ. Food and social classes in late Medieval and Renaissance Italy. In: Flandrin JL, Montanari M, editors. Food. A culinary history from Antiquity to the Present. New York: Columbia University Press; 1999. p. 302-312 9. Katzenberg MA. Stable isotope analysis: a tool for studying past diet, demography, and life history. In: Katzenberg MA, Saunders SR, editors. Biological anthropology of the human skeleton. New York: Wiley-Liss; 2000. p. 305-327. 10. Marchetti A, Pellegrini S, Bevilacqua G, Fornaciari G. Kras mutation in the tumor of Ferrante I of Aragon, King of Naples (1431-1494). The Lancet. 1996;347:1272. 11. Soussi T, Wiman KG. Shaping genetic alterations in human cancer: the p53 mutation paradigm. Cancer Cell. 2007;12:303-12. 12. Riera-Melis A. Society, food and Feudalism. In: Flandrin JL, Montanari M, editors. Food. A culinary history from Antiquity to the Present. New York: Columbia University Press; 1999. p. 251-267.

Paleonutrition and Paleopathology: Food ad Disease at the Renaissance Courts of Naples and Florence

FORNACIARI, GINO
2008

Abstract

Carbon and Nitrogen stable isotope analysis of bone collagen is at present an almost routine method in the study of ancient diets. By stable isotope analysis it is possible to determine directly the type of diet of an individual and consequently of a population group. Therefore, at regional levels, the similarities and differences in the diet of individuals within an entire population can be easily detected (Katzenberg, 2000). This is a preliminary report of a more complex study, still in course, of 25 individuals from the Basilica of S. Domenico Maggiore in Naples (15th-17th centuries) (Fornaciari, 1999, 2006) and of 20 individuals from the Medici Chapels of the Basilica of S. Lorenzo in Florence (16th-17th centuries) (Fornaciari, et al, 2007). The Italian Renaissance princes had a much wider choice of food than other classes (Grieco, 1999). For this reason their diet should not be considered typical of the wealthy class as a whole, in the same way as the isotopic data observed in the population groups sampled for this study may not be representative for all the groups of the elite Italian society. Historical records reveal that after meat and wine, which constituted the central nucleus of the nobles’ diet, eggs and cheese also appear, although much less frequently (Riera Melis, 1999). Th ese two types of food, with high protein content, were combined with meat on normal days and alternated with fish on penitential occasions. In the aristocratic dietary regimen, vegetables occupied a secondary place, with almost total absence of fruit (Grieco, 1999). The isotope values obtained are remarkably similar to those published for other late Medieval and post-Medieval populations in England and clearly refl ect the large intake not only of meat but also of marine foods by the Italian aristocratic classes, especially from southern Italy, in the 15th-17th centuries (Fornaciari, 2008). Other foods high in proteins and enriched in nitrogen, such as freshwater fi sh, birds (and their eggs), or mollusks are likely to have provided smaller contributions. Given the large variety of dietary resources attested by historical documents, this conclusion can hardly be surprising. I present three “clinical” cases, important from paleonutritional and paleopathological point of view. Th e left foot of Ferdinando I de’ Medici, Grand Duke of Tuscany (1549-1609), shows a scoped-out defect located at the peri-articular and articular surface of the interphalangeal joint of the hallux dorsum, with partial destruction of the sub-chondral plate; the lesion, revealing at X-ray an evident sclerotic margin, involves both the bones of the joint and is typical of chronic gout (Fornaciari et al, 2007). High values of δ15N demonstrate a diet very rich in meat from terrestrial animals. δ13C values show a minor integration with foods of marine origin (fi sh) (Fornaciari, 2008). This isotopic profile well correlates with the frequent attacks of gout referred by court chroniclers and with the diagnosis of chronic gout of the left big toe revealed by the paleopathological study. The skull of Don Filippino de’ Medici (1577-1582), eldest son of the Grand Duke Francesco I, reveals non-severe external hydrocephaly, as witnessed by the macroscopic and radiological study, and by a 16th century autopsy report. Rickets also worsened this pathological condition (Fornaciari et al., 2007). The δ15N isotope value of don Filippino is anomalous and intermediate between the very high levels of breast-feeding babies and those of post-weaning children. This value reveals a diet very rich in proteins of animal origin. If we plot the δ15N and δ13C values together, we can see that don Filippino shows a very anomalous and isolated position. Isotopic signals reveal a strong intake of terrestrial animal proteins, while the level of marine proteins falls within the normal range of the group (Fornaciari, 2008). We are in front of a really unbalanced diet of the child, with regard to either meat or milk. On the basis of the historical, paleonutritional and paleopathological data we can speculate about the life of don Filippino. The eldest son of the Grand Duke, the hereditary prince, was a frail and sickly child, with frequent indispositions and persistent fevers, aff ected by rickets and a light form of hydrocephaly. Probably for this reason, his parents and the court doctors worried about the child’s health, and forced him to eat more meat, considered at that time “the fi rst source of physical strength”(Riera-Melis, 1999). Th is fact would explain the presence of this isotopic signal in the bones of the small prince. Autopsy of the mummy of Ferrante I d’Aragona, king of Naples (1431-1494), revealed a moderately differentiated colon adenocarcinoma extensively infi ltrating the muscles of the small pelvis. Histology showed exceptionally well preserved neoplastic cells, disposed in cords and solid nests or forming pseudo-glandular lumina, disseminated in a fibrous tissue containing striated muscular fi bres. Th e cells appeared tall and crowded, with abundant cytoplasm and quite regular hyperchromatic nuclei (Fornaciari et al, 1993). The excellent, almost exceptional preservation of the histological structure of this unique tumor is probably due to the very dry microclimatic conditions of the church and certainly to the preservation substances used for embalming. Ancient DNA amplification of the neoplastic cells by polymerase chain reaction (PCR) in the mummy of Ferrante I evidenced a typical point mutation of the K-ras gene codon 12 (Marchetti, et al, 1996; Falchetti, et al. 2006). The recorded genetic change represents the most frequent mutation of the K-ras gene in sporadic colorectal cancer and is characteristic of the eff ects of a number of “alkylating” agents. Th e N-nitroso compounds (NOC) and their precursors, have been widely investigated for their diff usion and well known relevance in gastro-intestinal cancerogenesis. In particular the carcinogenic properties of N-nitroso compounds (NOC) are also associated with their ability to “alkylate” DNA. Recent studies have focused on the importance of endogenous NOC (Soussi, et al, 2007). The diet contains many constituents which can be nitrosated in the gastrointestinal tract (especially in the stomach). Increased intake of red meat induces a significant (3-fold) increase of faecal N-nitroso compound (NOC) levels, with a range of exposure in the faeces similar to that from tobacco-specific NOC in cigarette smokers. Th is endogenous formation of NOC in the human colon well explains the association between red meat and colorectal cancer (Fornaciari, et al, 1999). The portraits of Ferrante, both sculptures and pictures, reveal growing obesity from youth to maturity. Court chroniclers report on the strong preference of the king for red meat, especially game. Examination of the mummy of Ferrante, even if severely damaged by a fi re in 1509, confirms his obesity. Histology reveals severe arteriosclerosis of the carotid arteries. Finally, the paleonutritional data, with their high level of δ15N, show a massive intake of animal proteins (meat). The alimentary “environment” of the Neapolitan court of the XV century and the sovereign’s habits, with his abundance of natural endogenous alkylating agents, can well explain the K-ras mutation causing the tumor which killed the Aragonese king over fi ve centuries ago (Fornaciari, et al, 1999). In conclusion, the investigation into the diet of the Renaissance aristocratic classes in Italy well demonstrates the potential of the studies on Carbon and Nitrogen stable isotope analysis in archeology and paleopathology. REFERENCES 1. Falchetti M, Lupi R, Ottini L. Molecular analysis of a colorectal carcinoma from a mummy of the XVth century. Journal of History of Medicine. 2006;18:943-951. 2. Fornaciari G, Castagna M, Naccarato AG, Tognetti A, Collecchi P, Bevilacqua G. Adenocarcinoma in the mummy of Ferrante I of Aragon, King of Naples (1431-1494). Paleopathology Newsletter. 1993;82:7-11. 3. Fornaciari G, Marchetti A, Pellegrini S, Ciranni R. K-ras mutation in the tumour of King Ferrante I of Aragon (1431-1494) and environmental mutagens at the Aragonese court of Naples. International Journal of Osteoarcheology. 1999;9:297-304. 4. Fornaciari G. Renaissance mummies in Italy. Medicina nei Secoli. 1999;11:85-105. 5. Fornaciari G. Le mummie aragonesi in San Domenico Maggiore di Napoli. Medicina nei Secoli. 2006;18:875-896. 6. Fornaciari G, Vitiello A, Giusiani S, Giuffra V, Fornaciari A, Villari N. The Medici Project: fi rst anthropological and paleopathological results of the exploration of the Medici tombs in Florence. Medicina nei Secoli. 2007;19:521-544. 7. Fornaciari G. Food and disease at the Renaissance courts of Naples and Florence: a paleonutritional study. Appetite. 2008. In press. 8. Grieco AJ. Food and social classes in late Medieval and Renaissance Italy. In: Flandrin JL, Montanari M, editors. Food. A culinary history from Antiquity to the Present. New York: Columbia University Press; 1999. p. 302-312 9. Katzenberg MA. Stable isotope analysis: a tool for studying past diet, demography, and life history. In: Katzenberg MA, Saunders SR, editors. Biological anthropology of the human skeleton. New York: Wiley-Liss; 2000. p. 305-327. 10. Marchetti A, Pellegrini S, Bevilacqua G, Fornaciari G. Kras mutation in the tumor of Ferrante I of Aragon, King of Naples (1431-1494). The Lancet. 1996;347:1272. 11. Soussi T, Wiman KG. Shaping genetic alterations in human cancer: the p53 mutation paradigm. Cancer Cell. 2007;12:303-12. 12. Riera-Melis A. Society, food and Feudalism. In: Flandrin JL, Montanari M, editors. Food. A culinary history from Antiquity to the Present. New York: Columbia University Press; 1999. p. 251-267.
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