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This is a site-wide search. To search for specific collection objects, please return to the Collection Search page. Advanced Search. Back to list. One of a pair of unusually large views, depicting the Bacino di San Marco from opposing vantage points. It is however a clever composite image of several viewpoints within the courtyard of that church, and includes several campanili bell towers of churches that cannot be seen from that vantage point. The figures in the foreground represent different levels of Venetian society; from the seated beggar on the left, the merchants in the centre, and the priest and lawyer engaged in conversation on the right. There is the customary assortment of sea vessels in the picture, including a burchiello, or passenger boat, being towed in the middle ground. The triangle of the foreground terrace - framed by the temporarily-docked burchiello with the detail of passengers embarking - is matched by the boat in the middle of the painting. Its two masts are in turn replicated in the vertical soar of the Campanile di San Marco and the dome of Santa Maria della Salute. The boats are gently balanced in the lagoon and the whole composition is again framed, on the left hand side, by the profile of a vessel with its sail blowing towards the city. The tower collapsed in March the two pictures can thus be dated c. It is an attractive, clearly identifiable view of a type calculated to appeal to the Grand Tourist, with picturesque elements of local colour reinforcing the idea of Venice as an exciting cosmopolitan centre. Acquired at an unknown date by the 1st Marquess of Hertford, who went on the Grand Tour he was recorded in Rome in and Genoa in The Wallace Collection.

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Official websites use. Share sensitive information only on official, secure websites. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. In this paper, we present the results of toxicological analyses of preserved brain tissue and bone samples from the remains of the seventeenth century patients of the Ospedale Maggiore , the main hospital in Milan and one of the most innovative hospitals in Europe from the Renaissance period. Beneath it, the crypt functioned as the burial place for the deceased of the hospital. Anthropological, paleopathological, histological, radiological examinations and radiocarbon dating were also carried out. As a result, archeotoxicological analyses revealed the presence of codeine, morphine, noscapine and papaverine, derived from Papaver somniferum , a plant present in the hospital pharmacopeia used as a narcotic, analgesic, astringent, coagulant, and antitussive agent. In , Francesco Sforza, duke of Milan, founded the Ospedale Maggiore in the heart of the city, a major healthcare institution dedicated to the care of the poor social class of Milan. It became the main hospital of the city in the sixteenth century and a model during the Renaissance for similar institutions across Italy, and even Europe, because of its innovative and avant-garde free medical assistance to the poor, care of the ill and hospital organization. An improvement of the efficiency in healthcare was established by specialized therapies instead of general assistance, by employing specialized doctors and surgeons rather than generic nurses, by introducing daily medical rounds and by reforming hygienic conditions through the installation of an efficient sewage system and daily change of linens of the beds of the patients 1 , 2. Different chambers, created for the deposition of the bodies, were built under the floor of the crypt; inside these 14 chambers lie an estimated 2. The latter was an Austrian pharmacopeia readapted for Italian use reporting all plants and medical preparations mixture of plants and animal derivatives produced in the pharmacy of the hospital from the fifteenth century to the last days of the hospital in the early twentieth century. Moreover, inside the pharmacopeia are listed all the medical plants as well as therapeutic preparations used to cure the patients; furthermore, the part of the plant utilized i. Analytical and forensic toxicology when applied to ancient remains strengthen the field of archeotoxicology, allowing the detection of molecules present inside the human remains, thus implementing our knowledge on the history of medicine and disease as well as on habits and even environmental pollution. Indeed, the greatest challenge in archaeotoxicology is that it deals with complex and unconventional biological matrices that have been subjected to decomposition, diagenesis, taphonomic processes and very long post-mortem intervals PMI. It is thus essential to choose the most adapted biological matrices able to preserve molecules of toxicological relevance. The brain tissue is an organ often collected for toxicological investigation 3 that reflects the drugs present in the circulatory stream of a body at the time of death of a subject. The analytes can pass the blood—brain barrier remaining trapped inside the brain tissue at the time of death of an individual 3. Even the bone tissue constitutes an alternative biological matrix that can be used in archaeotoxicology for analytical investigation, considering that it is one of the only remaining biological matrices that can be found after a very long PMI 5. The arteries that enter, through the nutritive foramina, inside the bone tissue, transport the analytes in this matrix. The bone tissue then incorporates the molecules into the inorganic matrix through the bone remodeling cycle 6 , hence a fraction of these substances remains trapped inside the inorganic portion and can be detected after the death of the individual even after a very long PMI 5. Therefore, the bone tissue at times acts as shield for the analytes of toxicological interest preserving the molecules inside its inorganic portion until the time of analyses 5 , 7 — 9. As a consequence, the present study also aimed at a specific choice of biological samples and customized extraction procedures and chromatographic techniques which could reduce the limits imposed by diagenesis and taphonomy. In literature, many studies have shown that medication drugs and drugs of abuse can be detected in preserved tissues, hair and bones 10 — 12 and how to determine their presence in bones 5 , 7 , 8 , 10 , 17 — 20 , nails 21 — 23 , teeth 10 , 24 — 26 and hair 10 , 12 — 16 , In these studies, the detection of substances was limited to some drug categories, like antidepressants 7 , 17 , 20 , 21 , antipsychotics 7 , 17 , 20 , amphetamine 7 and methamphetamines 5 , benzodiazepines 5 , 7 , 18 , 20 , cannabinoids 5 , 7 , 10 — 12 , 26 , opioids 5 , 7 , 8 , 18 — 20 , 22 — 25 , non-benzodiazepine, sedative hypnotic 20 and stimulants 5 , 7 , 10 — 16 , 18 , 20 , 22 , either in single individuals 8 , 11 , 12 , 17 , 23 , 25 , or small populations 5 , 7 , 10 , 18 — 22 , 24 , Methods used for toxicological investigation have been primarily a GC—MS analyzer 10 , 12 — 14 , 16 — 19 , 22 — 26 and RIA 10 , 12 , 15 , 16 , followed by a more recent instrumentation, the LC—MS analyzer 5 , 7 , 20 , In one case, the biological samples were analyzed with fluorescence polarization immunoassay and gas chromatography-flame ionization detector 8. The literature review has been summarized in Table 1. To the best of our knowledge, there are no reports of opioids detected in biological samples with a post-mortem interval PMI of over 25 years or reports that compare the toxicological results with an extremely detailed pharmacopeia. Therefore, the present archeotoxicological study is the first to search and report active molecules from Papaver somniferum on ancient human remains and, in particular, on bone. It also is the first to associate substances reported in hospital records and pharmacological therapy to the remains of patients. All biological information obtained from anthropological and paleopathological investigations are reported in Table 2. The crania were all fragmented, mostly constituted of bones of the neurocranium, except for C3 and C5 which are almost complete with only part of the parietal and occipital bones missing, respectively. The bones were fragile, prone to fragmentation, and often covered with a white staining. Due to the absence of cranium in case no. For two crania C6, C7 , the determination of sex was not possible due to the taphonomic absence of the areas of the cranium needed for the analysis of the morphometric traits. For similar reasons, in four cases, the estimation of age-at-death could not be performed C1, C4, C6, C8. In the remaining four crania, the age was estimated 29 — 31 and revealed two young adults C3, C5 in which age could not be further detailed because of fragmentation, a 30—45 years old individual C2 based on palatine sutures 31 , one subadult 11—12 years old, C7 based on AlQahtani Population affinity could be estimated in two crania C4, C6 of European White ancestry through Hefner morphoscopic traits 32 , In spite of the commingling of the remains, C2 was anatomically associated to other bones right clavicle, right humerus and eight right ribs. Based on humeral bone length, stature was estimated at The cranium presented pathological signs of tertiary syphilis confirmed by CT analysis, with localized porosity and bone cavitation involving the outer table 34 , 35 Fig. Moreover, antemortem trauma on the left parietal bone was reported in C5 which could be the result of either sharp force trauma or trepanation following blunt force trauma. The lesion, Although some forensic cases of survival to sharp force injury to the head, where a blow has caused the loss of bone tissue and secondary radiating fracture lines, exist, a different and perhaps more solid interpretation would be that of a linear fracture of the left parietal bone through blunt force trauma then treated with a scraping trepanation followed by long term healing Fig. Ante-mortem trauma of the left parietal bone arrow , followed probably by trepanation and long term healing; b superior-lateral view of ante-mortem trauma on cranium MI CG 21 O-US Indeed, the calibrated results reported with Radiological imaging of the antemortem trauma discovered in C5 are reported in Fig. Signs of tertiary syphilis could be evidenced on CT imaging presented in Fig. The rest of the crania revealed no pathological or traumatic evidence. From a—h , left lateral view of cranium. From c—g , CT images of antemortem trauma probably followed by trepanation on left parietal bone. Different active principles of opium poppy Papaver somniferum were observed morphine, codeine, papaverine and noscapine both in preserved brain tissue and bone samples, as reported in Table 2. All the samples under investigation were screened with a customized inclusion list, containing the alkaloids of the medical plants listed inside the hospital pharmacopeia. The molecules under investigation were confirmed following the international standard guidelines for forensic toxicology Therefore, the analytes were identified considering the parent ion together with characteristic fragmentation of each molecule. The signal-to-noise ratio was above 3 for all the molecules, permitting the qualitative confirmation. Then, the confirmation was assessed via reference material analytical standards comparing retention time and mass spectral ion ratio. The identification criteria used for the identification of the analytes are reported in Table 3 and an example of chromatographic spectrum of each molecule are reported from Figs. Chromatographic spectrum upper lane and mass spectral ion ratio lower lane of morphine detected in C4. Chromatographic spectrum upper portion and mass spectral ion ratio bottom portion of codeine detected in PBT5. Chromatographic spectrum upper raw and mass spectral ion ratio lower portion of papaverine detected in C3. Chromatographic spectrum upper portion and mass spectral ion ratio bottom raw of noscapine detected in PBT1. The detection of papaverine, noscapine and codeine allowed to confirm that the plant was administered at the patients, considering that those three alkaloids can be detected only with the consumption of the Papaver somniferum , excluding any possible external and more recent contamination due to the heroin or morphine. Toxicology applied to historical and archaeological human remains is an innovative and relatively recent discipline, referred to as archaeotoxicology. This paper aims to contribute to this field of research by searching analytical signs of the administration of medical plants in hospitalized individuals of seventeenth century Milan. The results obtained on cranial and brain samples showed the presence of molecules, namely morphine, noscapine, papaverine and codeine, which can be attributed to the intake of the plant Papaver somniferum by the subjects included in this study. These results, to the best of our knowledge, constitute the first report on the detection of opium in historical and archaeological human remains. Indeed, according to the literature, this category of drugs has never been detected in soft tissues nor in bone samples. Some molecules, specifically cocaine, cotinine, and tetrahydrocannabinol were detected in ancient archaeological human remains 10 , 12 — 16 , 37 , even though the reliability of the interpretation of some of these works 10 , 38 have been questioned Indeed, Musshoff et al. However, no study has reported the detection of opium in ancient biological remains, whether they were bones, hair, nails, teeth, or preserved soft tissue, therefore we can consider this study as the first to prove the administration of opium in archaeological human remains of the Modern Age. In the present case, the archaeotoxicological analyses performed on preserved brain tissues and cranial samples of different deceased patients of the Ospedale Maggiore highlighted the presence of active principles attributable and specific to opium poppy Papaver somniferum , namely morphine, noscapine, papaverine and codeine, detected in both preserved brain tissues and bone samples Table 1. Specifically, noscapine, papaverine, and codeine active principles of Papaver somniferum were noted in preserved brain tissue, whereas in addition to these molecules, morphine was also detected in bone samples. The skeletons with traces of Papaver somniferum belonged to three females including two young adults , one male, and one subadult of 11—12 years for whom sex estimation could not be performed. In one case with positive toxicological findings no. C5 , signs of ante-mortem trauma were seen on the left parietal bone Figs. This could have been responsible for chronic inflammation and pain which may have been treated with the use of the Papaver somniferum plant at the hospital. Opium was listed in the apothecary's archives as early as in form of laudanum or black poppy seeds; in the inventory list of the presence of white poppy seeds, black poppy seeds, poppy syrup and thebaic opium is reported, while in the hospital also introduced laudanum patches The findings presented in this research therefore confirm the archival data and implement our knowledge of the history of medicine in Milan. Paleopathological signs of tertiary syphilis were found on cranium C2, yet no toxicological findings were obtained on either bone or brain samples of that individual; hence, no hypothesis regarding the administration of medical plants may be elaborated for this subject. From a toxicological point of view, the matrices investigated preserved brain tissue and bone samples can be representative of two different modalities of substance administration. On the one hand, preserved brain tissues, like all organs and biological fluids, represent an administration of drugs, even in single intake, near the time of death of a subject. On the other hand, the bone tissue reveals an administration or intake of the drug over time either occasional or chronic 5. Thus, from the toxicological investigation on preserved brain tissues, we can state that four individuals PBT1, PBT6, PBT8 and PBT9 had been given derivatives obtained from opium poppy slightly before death probably while they were at the hospital and that the drug they had received during hospitalization may have been administered because of their therapeutic properties, given their common usage at the hospital as narcotic, analgesic, astringent, coagulant, spasmolytic and antitussive In this paper, we presented in the context of an interdisciplinary study between archaeotoxicology, radiology, archeology, history of medicine, anthropology, and paleopathology, the presence of opium poppy in human remains for the first time in archaeological contexts thus implementing our knowledge of the medical and health care habits in the seventeenth century. Indeed, the analyses revealed the presence of active principles morphine, codeine, noscapine and papaverine that can be considered derivatives of Papaver somniferum. This plant was present in the historical archives of the pharmacopeia of the hospital, and was prescribed as a narcotic, analgesic, astringent, coagulant, spasmolytic and antitussive agent. Permissions to examine skeletal and brain remains in the present study were obtained from an agreement with the Sopraintendenza Archeologia, Belle Arti e Paesaggio della Lombardia , a regional institution of the Italian Ministry of Cultural Heritage, following the ethical protocol of the agreement itself. Eight crania were selected for study as they still contained preserved soft tissue. A ninth sample of preserved brain tissue was found during the excavation, but it was not possible to reassociate it to its cranium, therefore the soft tissue was sampled without the reassociation to the cranium of origin. These preserved brain tissues were found in situ within the vault of eight crania. All have similar consistency and appearance hence one was sampled for histological analysis. Macroscopically, the mummified brain tissue presents portions of cerebral hemispheres in which gyri and grooves can be easily recognized. In cross section, a thin outer layer of gray-black color was appreciated, attributable to the cerebral cortex, and below this layer was highlighted a gray-whitish area, ascribable to white substance. The areas collected are reported in Fig. After the histological processing, the samples were analyzed with optical microscopy Square named as 1 referred to the frontal cortex; square labeled as 2 was the occipital brain area, and square called 3 referred to parietal brain area. Light microscopy showed extensive decomposition artifacts, bacterial contamination and post-mortem lytic changes. The residues of cerebral parenchyma appears eosinophilic with a paucicellular component formed by a reticular matrix, crystal deposits and bacterial contamination. Bone samples from these crania were also collected. A total of eight specimens were sampled due to the absence of the cranium associated to PBT9. Brain samples PBT were numbered in the same order as their respective bone; therefore, we obtained a classification from C1 to C8, with the absence of C9, as the cranium of PBT9 was missing. The access to the chamber was only possible from an opening in the ceiling and the manhole had remained sealed until the first opening. At that time, the archeologists accessed the chamber equipped with personal protective equipment to perform the first site inspection. After the preliminary inspection, the first excavation campaign begun. The remains were retrieved by the archaeologists, equipped with personal protective gear and gloves, under the supervision of the toxicologists. The post-cranial bones associated to the cranium C2 were also collected together with the cranium and placed in a separate sterilized box, in order to permit the anthropologists to study the entire skeleton. The samplings performed for each investigation toxicological, radiocarbon, and histological investigations were performed with sterilized scalpel and handsaw. Well-preserved brain tissue samples were collected with a scalpel. The sample site was determined based on the consistency of the samples, choosing the best preserved area, as no specific zone of sampling is suggested for forensic toxicology specimen collection in such cases 3. For histological investigation, the section of sampling was selected based on the presence of well-preserved and well-visible encephalic convolutions, hypothesizing more preserved structures. For histological investigations three samples collected from the frontal, parietal and occipital brain area were selected. The bone sample for the radiocarbon investigation was collected based on its weight and preservation, indeed the sample should weight about 5 g and be well-preserved with the cortical bone not affected by diagenesis or taphonomic processes. For toxicological investigations, the cranial sample was collected considering that the cranium was the only bone detected in the individuals under investigation apart from C2 that was associated with post-cranial bone samples. However, to perform a standardized sampling, we decided to collect only cranial samples, even if other bones were present see post-cranial bone of C2. Moreover, the cranial sample can be considered a good bone matrix in forensic toxicology considering the excellent results obtained from previous studies that compared them with other bone sample sites 5 , The crania were cut with a sterilized handsaw on the occipital bone adjacent to the foramen magnum, and in cases where the foramen magnum was not preserved the parietal bone, in accordance with previous papers 5 , The crania are fragmented mostly constituted of neurocranial bones, except for C3 and C5 that were almost complete with only part of the parietal and occipital bones missing, respectively. The bones were fragile, prone to fragmentation, and often covered with a white adipocere-like staining. In spite of the commingling of the remains, C2 was anatomically associated to postcranial bones right clavicle, right humerus and eight right ribs. Due to the absence of cranium in the case no. The subjects were assessed for age-at-death, sex and ancestry estimation using standard anthropological methods. Specifically, sex estimation was based on the evaluation of sexual dimorphic features of the cranium in particular the anatomical points of the glabella, supraorbital margin, mastoid process, nuchal crest, and mental eminence Age-at-death was estimated based on epiphyseal fusion, dental eruption and stage of suture closure for adult individuals 31 , 46 — When possible, biological ancestry was determined based on the correlation of eleven morphoscopic cranial traits and their frequency in reference populations to assess population affinity Pathological signs were evaluated following standard methodology 34 , 35 , 52 — Radiocarbon investigation was performed to ascertain dating obtained by historical archives of the hospital. One bone sample, collected from cranium no. The Conventional Radiocarbon Age BP was calculated using the Libby half-life years , is corrected for total isotopic fraction and was used for calendar calibration where applicable. Results greater than the modern reference are reported as percent modern carbon pMC. Quoted errors are 1 sigma counting statistics. Calculated sigmas less than 30 BP on the Conventional Radiocarbon Age are conservatively rounded up to References for the calendar calibrations were Ramsey 56 and Reimer et al. The material was submitted to bone collagen extraction and the bone collagen was analyzed for the radiocarbon investigation. The pMC was The calibration was performed with BetaCal4. In addition, imaging was performed to compare and confirm the anthropological data and to obtain radiological assessment before the sampling of bones, a disruptive procedure necessary for toxicological investigations. Furthermore, radiological investigations could reveal the presence of pathology and perhaps the reason for administration of drugs. The crania were imaged using x-rays in frontal, lateral, superior, and inferior views, whenever possible. Two crania were also imaged with computed tomography CT to investigate the macroscopic signs of tertiary syphilis and antemortem trauma present on cases C3 and C6 respectively. Those crania were laid on the CT table on their skull base. A spiral acquisition was performed with the following parameters: kVp and 40 mAs; exposure time CT data were post-processed to obtain multiplanar reconstructions applying different filters using Horos v3. Samples were collected from the preserved brain tissues and crania. All biological samples were powdered in a ball mill Mixer mill MM , Retsch and 0. One hundred ng of Internal Standard SKF A Proadifen hydrochloride , obtained from the working solutions previously prepared and correctly stored, were added to 0. Considering the differences between the biological samples chosen, two different samples preparation were applied. For the preserved brain samples, 8 mL of pH 9 phosphate buffer solution were added, whereas cranial samples were supplemented with a water solution with EDTA at the Both solutions obtained were agitated on a Vortex mixer Heidolph, REAX top , placed on a rotating wheel Falc F for 48 h and then centrifuged for 30 min at rpm Thermo Scientific, Heraeus Biofuge primo centrifuge. Twenty mM ammonium formate in water and MeOH were the solvents that constituted the mobile phase for the analyses. The electrospray tension with positive mode and the positive ion spray voltage were set at 3. Sheath gas, aux gas and sweep gas has been set at 45, 20 and 10 Arb respectively. CID gas was 1. Approval to conduct this research was issued by the Sopraintendenza Archeologia, Belle Arti e Paesaggio della Lombardia , a regional institution of the Italian Ministry of Cultural Heritage, following the ethical protocol of the agreement itself. The data underlying this article are available in the article and from the corresponding author on reasonable request. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. This section collects any data citations, data availability statements, or supplementary materials included in this article. As a library, NLM provides access to scientific literature. Sci Rep. Find articles by Gaia Giordano. Find articles by Mirko Mattia. Find articles by Lucie Biehler-Gomez. Find articles by Michele Boracchi. Find articles by Stefania Tritella. Besta, Milan, Italy. Find articles by Emanuela Maderna. Find articles by Alessandro Porro. Find articles by Massimiliano Marco Corsi Romanelli. Find articles by Antonia Francesca Franchini. Find articles by Paolo Maria Galimberti. Find articles by Fabrizio Slavazzi. Find articles by Francesco Sardanelli. Find articles by Domenico Di Candia. Find articles by Cristina Cattaneo. Contributed equally. Received Sep 8; Accepted Jan 10; Collection date Open in a new tab. Publisher's note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. These authors contributed equally: Gaia Giordano and Mirko Mattia. Similar articles. Add to Collections. Create a new collection. Add to an existing collection. Choose a collection Unable to load your collection due to an error Please try again. Add Cancel. Antidepressants, antipsychotics, amphetamines, benzodiazepines, cannabinoids, opioids, stimulants. Benzodiazepines, stimulants, antidepressants, antipsychotics, anesthetics, opioids, non-benzodiazepine, sedative hypnotic. Benzodiazepines, opioids, methamphetamines, stimulants, cannabinoids. Tertiary syphilis on right parietal bone: caries sicca stages 1—3.

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