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2. The wound and the leg amputation of Giovanni dalle Bande Nere (1498-1526): life and death of a mercenary captain of the italian renaissance
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Ventura, Luca, FORNACIARI, G., Saragoni (Forlì), L., Ercolani (Forlì), G., LICATA, M., RUSPI, A., IORIO, S., LARENTIS, O., TESI, C., BENAGLIA, P., TOSI, A, ARMOCIDA, G., RONGA, M., ROSSETTI, C., GIUFFRA, V., BARTOLOTTA, E., MILANESE, M., BOANO, R., PALLECCHI, S., SANTORO, E, BERTOLINO, S., VELLONE, V.G., BUFFELLI, F, FULCHERI, E., ALABISO, A., BRIDELLI, M.G., MAZZOTTI, M.C., FERSINI, F., PELOTTI, S., LUISELLI, D., TRAVERSARI, M., FUSCO, R., CERMESONI, B., RAVAGNAN, A., BADINO, P., DI CORCIA, T., DE ANGELIS, F., ROMBONI, M., VELTRE, V., LABARGA, C. MARTÍNEZ, RICKARDS, O., VARANO, S., CALDARINI, C., PANTANO, W., CATALANO, P., MARTÍNEZ-LABARGA, C., Rabino Massa (Torino), E., Licata (Varese), M., GUERRIERO, M., COLASURDO, F., POLLIO, A.M., PONZIO, G.V., CILLI, E., LUISELLI, D, GRUPPIONI, G., SARAGONI, L., PETRELLA, E., VENTURA, L., RIPANI, M., GAETA, R., MINOZZI, S., FORNACIARI, A., RICCOMI, G., LOMARTIRE, S., PICCHIONI, L., IOANNUCCI, M., PANELLA, S., MICARELLI, I., PAINE, R.R., TAFURI, M.A., MANZI, G., BUFFELLI, F., BONSIGNORE, A., CILIBERTI, R., Traversari (Forlì), M., Gaeta (Pisa), R., SACCHERI, P., TRAVAN, L., GOTTARDI, G.M., FOSCATI, A., FORTUNA, S., CARLI, A., PIOMBINO-MASCALI, D., BIRKHOFF, J.M., CISINI, S., DE STEFANO, F., RABINO, E., FERRARI, L., PULCINELLI, D., RULLO, D., LOMBARDI, A., METOVIC, J., PAPOLA, F., Ferrari (Asti), L., Fulcheri (Genova), E., PIETROBELLI, A., MARIOTTI, V., FUSARI, S., BETTUZZI, M., MORIGI, M.P., BELCASTRO, M.G., TONINA, E., AMARO, A., CAVALLINI1, L., COSCHINO, F., VERCELLOTTI, G., DE SANCTIS, M., ISHAK, R., ARINGHIERI, G., BRUSCHI, F., MANNINO, M., PANGRAZZI, C., GORINI, I., PAUDICE, M., BIATTA, C.M., PEÑUELA, L., and CINTI, A.
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Meeting Report ,Lecture ,Article ,Pathology and Forensic Medicine - Abstract
The tomb of Giovanni and his wife Maria Salviati at Cappelle Medicee in Florence was explored to investigate the skeletal remains. Anthropological and paleopathological examination defined: age at death, physical constitution and activity, skeletal diseases. The stump of the right leg was studied macroscopically, under stereoscopic microscope, at X-ray and CT scans, to detect type of injury and level of amputation. The study of the skeleton of Giovanni revealed that he was a vigorous man, 1.78 m tall, with an athletic body, estimated skeletal age of 25-30 years, medium-sized skull, narrow nose and great skull capacity (1494 cc). His well-developed upper limbs muscular insertions (deltoid, great pectoral, great dorsal, biceps, forearm muscles) and thigh muscles confirmed his great physical strength and robusticity. Strong hypertrophy of rotator cuff, great dorsal, teres minor and anconeus insertions were all present, as well as gluteal insertions to the femur, confirming he was a highly skilled horseman. The presence of numerous Schmorl’s hernias and a wedge partial collapse, with right spondylolysis, of the fifth lumbar vertebra, revealed that Giovanni had carried heavy loads since adolescence due to horse-riding and body armor. Diffuse bilateral enthesopathies were found at the clavicular insertions of deltoid and pectoralis major, as well as at the small trochanter (psoas muscle). Skeletal markers left by habitual horseback riding were all present: exostoses and ovalization of acetabula, hypertrophy of femoral rectum muscle, strong hypertrophy of the femoral biceps, great adductor, small and great gluteus, Poirier’s facet. Paleopathological investigation showed the aftermaths of several injuries: fractures of nasal septum and proximal third of the left humerus, injury from blade affecting right ulna and radius and swelling of the posterior surface of the right tibia, with underlying osteomyelitic focus in reparative phase, as well-documented on CT. The amputation level was exactly assessed: the tibia was sawn immediately below the proximal half of diaphysis and only the lateral portion was surgically treated with an horizontal cut. Only oblique splitting was found at the medial site of the tibia. At stereoscopic microscope, surgical section revealed a marked proliferation of endosteal callus, due to a previous harquebus shot injury occurred about one year before the death. Distal extremity of fibular fragment showed an oblique splitting and a horizontal cut, with no sign of reparative process in the medullar canal. Considering the morphological aspect of the tibial and fibular injury, it was due to a cannonball from a falconet of caliber 6-7 cm, as written by Benedetto Agnello in the same day of injuring. The limb had been severely damaged by a traumatic hemi-amputation when surgeon Abramo performed the intervention, consisting in a simple completion of the amputation and regularization of proximal fragments. In conclusion, paleopathological investigations lead to exclude the hypothesis of an amputation above the knee, since the surgeon Abraham performed the procedure as best as he could in conformity with surgical knowledge of that period., Biomechanical and kinesiological reasoning allow us to investigate traumas in Paleopathology. The focus of our analysis is to reconstruct pathomechanics, treatment and gait of a subject from the Late Medieval femur, which presents an important bone callus (1). The femur was discovered in a funerary crypt of the Sanctuary of Sacro Monte (Varese, Northern Italy), an important archaeological context inserted into the UNESCO heritage since 2003. The femur was studied with computer tomography and the reconstruction of the static and dynamic fictional outcomes of the lesion was performed by the Observation Gait Analysis (OGA). The OGA is the computerized analysis of the gait. This technique permits to observe movements of each articulation in the space, the posture and the gait underlining a movement strategy. The femur presents an important callus at the middle third proximal of the shaft. The fracture is oblique and caused by a direct trauma probably associated with occupational activities. The alignment of the segments in the frontal plane leads us to assume that the fracture was treated and the femur was immobilized with splints. The use of OGA allows us to understand the subject’s kind of gait after healing. Our analysis demonstrated that there was no reduction in bone mass. The deposition of new cortical bone near the fracture determines that the individual has gradually resumed loading the leg and was walking although with significant effects on posture and movement., In 2008, archaeological excavations carried out in the inner courtyard of the former Jesuit College of Alghero brought to light the San Michele cemetery. Characteristic of the site are some burial trenches, narrow and long pits containing the remains of 10 to 30 individuals, and some multiple tombs, which can be related to the plague epidemic that ravaged the city in 1582-83. The present study is focused on the demographic analysis of the 16 trenches containing 185 individuals and of one multiple tomb (T.141) with 14 individuals. In the case of the trenches it was possible to determine the sex of 178 individuals: 37 are males (20.7%), 53 females (29.8%) and 88 of undetermined sex (49.5%). As for the first two groups, 35.6% of individuals has an age comprised between 20 and 29 years; the two age ranges 30-39 and 40-49 years present the same percentage (25.6%), 8.9% are aged between 17 and 19, and finally 4.4% are over the age of 50. The undetermined sex category is represented by 81 subadults and 7 adults. As for the subadults, the most representative age group is that between 7 and 12 years old with 39.8%, followed by the 23.9% between 13 and 19 years, 18.2% between 2 and 6 years, and finally, 10.2% between 0 and 1 years. The multiple tomb 141 includes an adult individual (20-29 years) of undetermined sex, a woman of about 17 years and with a 35-week fetus in her womb, and finally 12 subadults in an age range between 0-1 years (21.4%), 2-6 years (7.1%) and 7-12 years (50.0%). The cemetery of San Michele presents some similarities with the French cemetery of Martigues struck by the plague in 1720. Also in this cemetery this type of trench burials was found, 5 in this case, with 199 individuals. The comparison between the paleodemographic curves for both cemeteries evidences a similarity of the mortality trend. The difference between normal and catastrophic cemeteries, related to a severe epidemic event, consists in the fact that while in the former there is a greater presence of infants and elderly, in the latter there is a certain homogeneity of mortality, proof of the fact that the plague kills in a random way and therefore all individuals present the same risk of death., Ossification of spinal ligaments can persist on skeletal remains requiring a differential diagnosis between diseases that can lead to spinal column ankylosis. The most relevant diseases that can lead to this condition are represented by ankylosing spondylitis (AS) and diffuse idiopathic skeletal hyperostosis (DISH). The differential diagnosis between these two conditions is discussed in two cases of our observation. Case n. 1: Policastro Bussentino, US 112 - The burial (US 112) was found in the convent of San Francesco in Policastro Bussentino, during an emergency excavation carried out in 2015. The skeleton on a stratigraphic basis is dated between 1846 and 1892. The skeleton is affected by the fusion of 12 vertebrae of the vertebral column, from the fusion of some costovertebral joints and of the sacroiliac joints. Case n. 2: Turin, Collection Marro, skeleton n.16809 - The skeleton belongs to the Egyptian Collection of the Museum of Anthropology and Ethonography of the University of Turin. It was collected during the archaeological campaign at Gebelein (Upper Egypt) in 1920 and dates back to the First Intermediate Period (2150-1990 BC). In this case, the pathology involves only the vertebral column in which the fusion of numerous vertebrae is found. AS is a progressive inflammatory disease of unknown etiology primarily affecting the diarthrodial joints of the spine, the costovertebral joints and the sacroiliac joints and usually begins in the second or third decade of life. First it affects the lumbar spine and the sacroiliac joints and progressively ascends until the entire spine and all costovertebral joints are affected. The result of this ankylosing process is the rigid, so-called bamboo spine with loss of its physiological curvatures. DISH is an ossifying diathesis producing ankylosis of the spine due to ligament ossification without intervertebral disk disease. It is not a true arthropathy because neither cartilage nor synovium are involved. It is rarely detected before the age of 40 years and the cause is unknown. The two pictures macroscopically can be confused because both involve the spine with ossification and ankylosis of it. The differential diagnosis on skeletal remains can be based on extimated age and sex and on the elements involved. AS occurs in the second or third decade of life and mainly affects men. DISH, instead, appears not before the fourth decade of age. DISH only affects the spine, while ankylosing spondylitis also affects the cost-vertebral joints and the sacro-iliac joint. The ligaments involved in the fusion of the spine are different in the two cases. DISH does not affect the intervertebral discs unlike AS which instead involves them and thus has a total fusion of the vertebral bodies. Although DISH and AS manifest in a similar manner, they are separate diseases. Both pathologies are quite common in mild and initial forms, but are rather rare in the full-blown, severe forms, which involve the whole spine. This topic appear poorly covered both in paleopathological and clinical literature, mainly consisting in small series and case reports. As a consequence, a wide inter-individual variability is present and only rarely an accurate report of the different involved ligaments is provided. In most of the cases the description is limited to a generic attribution to spinal ligaments. Differential diagnosis may be challenging if limited to anthropological examination of the skeletal remains and further radiologic and genetic tests are necessary to confirm our findings., The object of this study is the so-called “green mummy” of Bologna, a naturally mummified body that was found in the basement of an ancient mansion in Bologna in the 1920s of last century. The hard and soft tissues of the body are green for the most. They were analyzed by FTIR spectroscopy with the aim to gain information about the biochemical degradation process, to explain both the origin and the nature of the green color and to understand how it affected the body’s preservation. The corpse was found in a copper or bronze cist that, at the time of recovery, was closed with a lid but broken at the base. It is reasonable to assume that the body had undergone the processes of putrefaction, liquefaction, skeletonization and mummification within the container. Probably, the semi-fluid mass of water and decomposing substances came out from the cist through the break at its base; therefore, the not yet putrefied tissues exposed to the atmosphere of the basement, desiccated and mummified. The acidic liquids originated by the decomposition caused the container corrosion, favouring the release of copper ions. Metal ions both inhibited the enzymes responsible for chemical reactions in the early stages of the decomposition process and acted as biocides of microorganisms involved in decomposition. FTIR analysis of soft tissues, with and without green color, showed that tissues lacking the green patina were the best preserved revealing the protein structure only minimally deteriorated, contrarily to what was observed in green areas. We have hypothesized that copper ions might have caused the decarboxylation of the RCOO group of polypeptidic backbone favoring protein degradation. Therefore, copper did not favor the mummification process, which was probably due to the environmental conditions, such as the low temperatures typical of the basement and the low availability of oxygen. Due to the known biocidal action of copper ions, we assume that copper ions might have damaged dead tissue cells in the same way they damage the cell membrane of microorganisms with which it comes into contact, causing their death. After the body decomposition liquids came out from the container, copper corrosion products precipitated as copper compounds giving the remains the green color. On the soft tissues two mineral forms of copper have been found: copper carbonate and copper phosphate called malachite and sampleite. Also, FTIR analysis of bones revealed that copper did not contribute to tissue preservation. The measured mineralization index showed the alteration of both collagen and hydroxyapatite of colored bones unlike the colorless ones. A compound known as pseudomalachite was identified in the green bones, a form of hydroxyapatite where copper replace calcium. This substitution is responsible for the unusual green coloration of the bones., The new paleoradiological investigations carried out on the Egyptian mummified human remains, a head, a left hand and foot, housed in the Civic Archaeological Museum of Erba (Como, northern Italy), allowed us to acquire comprehensive digital images and to study the anthropological and paleopathological data of the subjects. The first macroscopic investigation revealed the presence of blue-glazed Faience tubes, adherent to the dorsal wrappings of the foot. This custom was particularly attested in the 26th dynasty, even if it appeared already from the 21st dynasty and seemed to continue until the Ptolemaic period. The tomographic analysis revealed the non-compatibility of the three mummified parts to a single individual, based on the different degrees of bone development and degeneration. The radiological investigation allowed us to acquire data on the embalming techniques applied, and on the health status and pathological conditions of the head. In particular, the parietal bones of the cranium exhibited two symmetrical areas of thinning and resorption of the outer table, which suggest a case of “biparietal thinning”, also known as “biparietal osteodystrophy”. The areas involved are well-circumscribed and elliptical in shape, localized between the obelion and the superior linea temporalis. Macroscopically, these areas are noted as slight depression and flattening of the outer cortical layer. The CT imagings also revealed the mature age of the individual, which, according to several studies, is compatible with this finding. A research in the literature, showed a prevalence of this condition in Egyptian individuals, associating it with different definitions and causes. Although the etiology of this affection is still not well-known, here the embalmed head of an elderly individual revealed the presence of biparietal affection, with thinning and resorption of the outer table, adding additional evidence of this finding in an Egyptian subject., The GEDEON project will allow us to broaden the knowledge about the human adaptation against changes in nutritional resources consumption. The main purpose of the project is to investigate the putative role of selective pressure that may have acted on specific genetic markers linked to changed dietary conditions. In order to reach this topic, the project aims to connect the scientific evidence obtained through osteological studies of the ancient remains dating across the major nutritional transitions, with the evaluation of genetic markers involved in metabolic pathways that may have been affected by nutrient bio-availability. The whole genome analysis of ancient selected skeletal specimens will be sequenced to compare the data obtained with those from extant people suffering of nutritional impairments, whose information on dietary requirements is available. Known polymorphisms that are classically referable to diet-derived homeostatic alterations will be selected, such as rs2066844, rs2066845 and rs2066847, whose association with inflammatory intestinal diseases (IBD) is well known. Other variants mapping on NOD2 gene (for example rs2066843 and rs2076756) will be focused, but they will represent only the starting point for the identification of causative molecular pathways modifiers. Furthermore, the markers with a well-known association with alterations such as celiac disease (CD) and primary hypolactasia (PH) will be also considered: PTPN2 and IL18RAP loci as well as various HLA system factors and the LCT gene will be primarily evaluated. This shortlist selection will constitute the beginning for the identification of several new markers to shed light on human genetic adaptation to the changed environmental conditions including the nutritional requirements., The application of the next generation sequencing techniques to the study of ancient DNA represents an outstanding improvement for clarifying complex scenarios related to genomic-based physio-pathological conditions, whose identification in ancient remains can be tricky. Indeed, the sole presence of osteological markers could be misleading for proper diagnosis due to the non-specific nature of such lesions. A thorough molecular evaluation has been performed on a skeleton of an adult woman dating to the Roman Imperial Age. The erosive and osteolytic markers located in the tarsal bones have led to hypothesize that she probably suffered from gout. In an attempt to integrate the differential diagnosis based on osteological data, whole genome sequencing analysis was performed. The bioinformatics pipeline identified the presence of two variants in the TSC2 gene, that is known to be associated with a rare genetic disorder, the Tuberous Sclerosis Complex, featured by signs that could be shared with those due to gouty arthritis. The application of these ultimate molecular techniques surely represents a successful diagnostic tool for the identification of genetic related disorders that could be only hypothesized in ancient times., Domenico Petruccelli (Raffaele da Sant’Elia a Pianisi), servant of God, was born in Sant’Elia a Pianisi (Campobasso) in 1816 in a family of honest and wealthy peasants. He lived in different convents between Puglia and Molise. Wherever he went, he won the esteem and veneration of everyone, so much so as to be called the “Holy Monk”. He died in the convent of Sant’Elia a Pianisi in 1901. In the same convent another famous monk arrived in 1904 fr. Pius from Pietrelcina (better known as Padre Pio), here the young monk breathed the aura of holiness left by the Holy Monk who died a few years earlier, drawing strength and inspiration. A first exhumation had been performed in 1934 and the skull was placed in a metal case. In 2017, 200 years after his birth, during the second exhumation and canonical recognition of the mortal remains, the skull was in good condition of conservation. It was covered in a thin layer of yellowish-white powder and traces of deteriorated fabric which formed part of the inner lining of urn. Cleaning operations The skull was cleaned. During this operation, we proceeded to collect in a container of numerous fragments adhered to the outer and inner surface of the skull. Consolidation operations The skull was impregnated with a thin protective membrane consisting of a very thin film of an acrylic resin (Paraloid B72TM) which protects it and consolidates its most delicate and fragile parts. Paleopathological study The alterations and lesions found are: in the left orbit, two holes (of undetermined origin) in the left parietal and temporal bones, in the right parietal bone, at the base of the skull, the jaw is absent. Attribution of sex We highlight a series of morphological findings characteristic of the male sex. They are: protruding glabella and sloping forehead; Rounded, wide and thick super-margin; Wide zygomatic arch that extends beyond the external acoustic meatus; Mastoid process robust, big and verticalized; Necked crest marked, wrinkled and very evident. Diagnosis of age Coronal, sagittal and lambdoidal sutures were considered, in accordance with the criteria of Acsàdi and Nemeskèri (1970) modified by Masset (1989). The endocranial closing index can be estimated at around 4, so the subject’s age is between 50 and 80 years, according to the known age of Father Raffaele of 85 years at the time of death. Craniometry The craniometric assessments carried out highlighted: a small, rounded, long, narrow, medium-high skull with rounded sagittal contours, oval and angular orbits, mean interorbitary distance, narrow nasal opening. All these characteristics allow us to state that the skull belonged to a Caucasian subject. Histological examination The fragments examined were composed of human organic material. These are extensively necrotic and rotten tissues. An important result has been achieved with the finding in the histological findings of many fungal hyphae, better highlighted with histochemical stains PAS and Grocott., Historical information related to the life of St. Mercurialis is very scarce, everything we currently know, we owe to its legenda, is contained within the manuscript Casanatense 718 dating to the 12th century. The only certain historical information concerns the ordination of one of his successors, Grato, which took place in Ravenna during the 5th century. The relics of St. Mercurialis, preserved inside the homonymous abbey, in the Cathedral of Santa Croce and in the Santissima Trinità church, during 2018 were object of the sixth canonical recognition, necessary to verify the state of conservation of the bones. Preliminary studies were performed by direct anthropological and radiological analyses by CT scan, FTIR analysis, ancient DNA and radiocarbon dating. St. Mercurialis, was about 1.60 meters tall, the age at death is 45-50 years, and he was not particularly robust, even if marked by repeated musculoskeletal stress probably linked to habitual activities such as walking and weightlifting. He suffered from osteoporosis and perhaps had some discomfort with the shoulder girdle. He had a deviated nasal septum from birth, a condition that perhaps caused him disorders such as sinusitis. He did not suffer from osteoarthritis and he had no particular indicators about deficiencies suffered during the first and last period of life. Analyses did not reveal indicators due to traumatic events and probably did not die by strangulation, as the hyoid bone was intact. FTIR analysis was carried out on the brown substance that partly covered the lower skeleton district, shows the typical spectrum of clayey materials. The characteristic bands reveal the presence of aluminum and silicon in greater quantities, and of other elements in smaller quantities. The clays constituting the soil can therefore be considered essentially illites containing kaolinite, smectite and quartz. This aspect confirms the numerous historical information concerning the floods suffered by the abbey. A patina that covered some bones was also detected. The obtained spectrum presents the typical absorptions of the vibrations of the hydrocarbon radicals CH2 and CH3, in addition to the intense absorption typical of the C-O-C group characteristic of carbohydrates. The characteristics of the spectrum therefore seem to be typical of a methylcellulose preservative. Radiocarbon dating and accelerometry mass spectrometry (AMS) dated the relics to the I-III century AD, an interesting date that is chronologically before the only historical indication we have about the life of St. Mercurialis and which collocates his episcopate into the first stages of evangelization of the Emilia Romagna Region. The preliminary analyses of ancient DNA were targeted on the hypervariable region 1 (HVR-1) of the mitochondrial DNA (mtDNA) and on Short Tandem Repeats (STRs) of the Y-chromosome and highlighted a rather pronounced diagenesis of the DNA. The subsequent analyses will be targeted to the capture of the entire mtDNA, coupled with next generation sequencing., Andrea was a Catholic priest and a member of the Order of Saint Augustine. He was born to a modest family in the village of Mascioni (northern borders of L’Aquila province) and as a child he worked as a shepherd. Around 1417 he met Augustine from Terni, Prior of the Augustinian convent in the near Montereale, and asked to enter their ranks to be ordered as a priest at the age of 25. He earned a bachelor and master’s degree in theology, becoming professor in Siena (1443), and Provincial Prior of the Umbria region. He also served as a travelling preacher in Italy and France, reforming several Augustinian monasteries in Umbria. He died on 18 april 1479 in Montereale and, according to hagiographies, his body was exposed without balsams in the conventual church for 30 days, giving off sweet odor and performing miracles. Subsequently, it was placed in the Choir until 1568, when it was displayed beneath the main altar. His beatification was celebrated by Pope Clement XIII on 18 february 1764. In 1787 the body was translated into a newly built repository, inside the new chapel dedicated to him. Documented Canonical Recogntions took place in August 1786, July 1943, June 1961, and between June and July 1989. The last was performed by one of us (MR) and the late professor Giulio Marinozzi. External examination of the body allowed to recognize a partially skeletonized mummy belonging to an old male subject (more than 70 years of age at death) and measuring 164 cm in length. The face was almost entirely covered by mummified skin, with traces of hair in the perioral region, chin, cheeks and anterior neck, according to devotional representations of the Blessed as a bearded elder. Soft tissues of forearms, hands, legs and feet appeared in a good preservation state. A large bone defect was observed in the occipital squama. Large skin cuts were observed in the anterior neck and left hemithorax. The ventral portions of the left ribs appeared cut and displaced within the thoracic cavity. Cut marks were also found on the left margin of the sternum body and on anterior branches of the pelvis. Preserved skin was observed only in the right hemithorax. No traces of internal organs were found in thorax, abdomen and pelvis. Moderate-marked osteoarthritis of the spine was noted. Unfortunately, a radiologic investigation of the body was not performed. After external examination, conservative treatment was performed. The body of Andrea revealed indisputable evidence of artificial mummification, representing the eleventh described case of an embalmed Saint or Blessed in Catholic Religion. Nine of these artificial mummies were created in central Italy (Umbria, Toscana, Abruzzo, Lazio) between the XIII and XV century. The employed evisceration procedures appear somewhat rough, without the complexity observed in other examples. It is worth to note that Montereale is located not far from L’Aquila and on the main route towards Cascia and Spoleto. The embalming of the Blessed Andrea took place only 35 years after the death and embalming of Saint Bernardino da Siena in L’Aquila and represents the second case in Abruzzo region., Liutprand, one of the greatest Longobard sovereign, was born in the early 90s of the 7th century and died in 744 at the age of about 55 years. According to the Historia Longobardorum of Paolo Diacono, he was King of Longobards from 712 to 744. The remains of King Liutprand suffered from many translations in the centuries and this is the main problem for the validation of the authenticity of the bone remains. The first grave was in the chapel of Sant’Adriano in the Longobard cemetery of Santa Maria in Pertica (Pavia). Later, in the second half of 12th century, the body of Liutprand was translated in the Basilica of San Pietro in Ciel d’Oro and located in a monumental mausoleum. New translations took place after the Council of Trento and in 1895, when the bones were placed in a niche in the floor of the church where they were found in January 2018. The bones, contained in a wooden box, appeared extremely fragmented and in a poor state of preservation. Anthropological examination highlighted the presence of bones attributable to three individuals. Most of the remains belongs to a robust male individual with strong muscular insertions, with an age at death between 40 and 50 years. There is also a second older male with strong muscle insertions, and a third adult individual of similar size. In fact, there are a pathological left tibia and some fragments of its right controlateral, and other fragments attributable to two other left tibiae. The tibia with pathological alterations presents the upper third of the diaphysis completely altered and enlarged by bone thickening due to a severe form of osteomyelitis. Bone repair is evident and the presence of a circular depression with a diameter of about 10 mm at the point of maximum thickening could represent the trace left by a pointed object that caused the perforation of the bone and the subsequent infection with osteomyelitis. The reparative process has however led to the healing of the lesion before death, which should have occurred not far from the event (maximum 2 years). Imaging studies (CT and radiological) on the skeletal remains were performed at the “San Matteo” General Hospital in Pavia. 14C dating provide a range from 430 to 640 for the first subject, 600-770 for the second and 530-670 for the last male. Isotopic data show a rather high nutritional status for the time, with a varied diet rich of meat. In conclusion, currently it is not possible to accurately define the identity of the three individuals for lack of archaeological data and for the fragmentary nature of the bones. The age of the subjects, the robust constitution and the nutritional data suggest a belonging to a high social class perhaps devoted to war activity. Future molecular studies may perhaps reveal a possible degree of kinship between the individuals and clarify the identity of the subjects., Born to one of the leading families in Besançon, Jean Bassand was a French Christian monk. After his initial profession in the Augustinian house of Saint Paul, he joined the Celestines (a branch of the Benedictine Order) in Paris, and subsequently became prior in the city of Amiens. The Celestine monks of France were a self-governing province of an Italian Benedictine reform of the late XIII century that no longer exists. They had a great influence, representing one of the most prominent observant groups in France, and an inspiration for reform movements across multiple orders. Jean Bassand represented the most important figure in the French Celestine congregation between XIV and XV century, being elected provincial prior on five occasions. He made great efforts to establish new Houses in the French province as well as abroad. The English King Henry V invited him to found and direct a friary in Isleworth near Sheen (now Richmond, London), whereas Martin I of Aragon asked him to establish the congregation in Barcelona. In 1443, he went to L’Aquila by order of the Pope Eugenius IV, to reform the monastery of Santa Maria di Collemaggio. He had troubles in this task and retreated to Rome arguing that the Aquilans were “difficult men”, but the Pope sent him back until his mission was accomplished. He died in L’Aquila on 26 August 1445. His body, covered with lime to be displayed, was found intact 18 years after. Since his death the mummy of Jean Bassand used to be kept in the Basilica of Collemaggio. After the major earthquake that struck down the city in 2009, his remains were recovered from the church to be kept in a secret location. Recently, an inspection of the body took place as a preliminary step of a forthcoming Canonical Recognition. The mummy appeared still fully dressed, with face and hands uncovered. The skin surface was extremely well preserved, and oblique illumination disclosed multiple, round, well-circumscribed plaques on the forehead, cheeks, and upper lip. Careful examination of the digital pictures enabled us to recognize at least 19 lesions. From a modern clinical viewpoint, the facial skin eruption of Jean Bassand meets most of the diagnostic criteria for multiple seborrheic keratoses. The age at death and the lifestyle of the Blessed, with frequent, long-distance travel under severe conditions, are fully compatible with this diagnosis. It is well known that male sex, increased age and sun-exposure may predispose individuals towards the development of these lesions. This case might represent the first ancient seborrheic keratosis described in the literature, although further analyses (external examination, computed tomography, histology) are needed to confirm the diagnosis. Advanced investigation methods might also enable us to understand if an ancient case harbours the same genetic mutations detected in modern patients. In this particular case, the treatment of the corpse with lime referred to in the ancient literature may have preserved the skin lesions by dehydration., This study presents the skeletal evidence for scurvy among 22 juveniles from the Longobard necropolis of Selvicciola, Italy (VII-VIII centuries AD). The paleopathological analysis revealed a combination of bilateral porous and proliferative bone lesions affecting the orbital roof (i.e., cribra orbitalia), the cranial vault (i.e., porotic hyperostosis) and some specific areas of the entire skeleton (i.e., sphenoid, hard palate, and scapula). This pattern is typical of infantile scurvy (Geber & Murphy, 2012). The investigation also revealed a bone tissue resorption of the anterior and lateral surface of thoracic and lumbar vertebral bodies and rib lesions in 17 of the 22 juveniles that showed signs of scurvy. This kind of skeletal manifestation associated with endocranial alteration and diffuse periosteal new bone formation (PNBF) is often linked to atypical or early-stage tuberculosis (Spekker et al., 2012). Our assessment suggests that the diet in Selvicciola was mainly deficient in vitamin C, causing scurvy. Among other problems, this might indicate poor sanitary conditions and further factors related to local environment and general state of health. Consequently, scurvy may have reduced the immune resilience of the juveniles leading to the development of TB in many of them (Miladinović-Radmilović & Vulović, 2015). Additional assessment via isotopic studies of the Selvicciola burial collection (Tafuri et al., 2018) indicates that the consumption of animal proteins in their diet was quite high. Hence, TB may have originated as Mycobacterium bovis (Roberts & Buikstra, 2003), coming from dietary consumption of infected animals. The preliminary macroscopic investigation of these subadult skeletons reveals lesions that suggest a complicated relationship among several factors influencing the health of these children. Diet, infection and lack of vitamins may have contributed to the poor health and death of these juveniles. Hence, showing a complicated situation in which, these children lived and dead in. This anthropological work illustrates how paleopathology can be used to interpret the health status of individuals of past communities even when the lesion evidence suggests multiple causal factors associated with death., The Institute of Clinical Pathology currently keeps the autopsy registers from 1891 to the present; the oldest one consists of some volumes in which the protocols and the related diagnoses were written by hand; for each case, a succinct clinical history preceded the external examination. The case presented here concerns obstetric pathology related to maternal post-partum death due to uncontrollable bleeding. The autopsy was carried out in 1892 on a woman suffering from rickets with scant muscular masses; the autopsy was performed 37 hours after the patient’s death. At that time (specifically from 1886 to 1905) the Director of the Unit of Clinical Pathology was Professor Vincenzo Brigidi and the autopsy room was situated near Pammatone Hospital, which was built in the district of Portoria and almost five centuries earlier, in the fifteenth century. This was the main hospital in Genoa, and played a fundamental role in local public health; the whole institute consisted of a single autopsy room. Full-blown or paucisymptomatic rickets was common in the Italian population of the nineteenth century and the first half of the twentieth century. The high incidence of this disease was due to deficiency factors or chronic nutritional stress, which led to forms of pseudo-rickets or latent rickets; this general situation required strict supervision of pregnant patients in order to avoid fatal intra-partum accidents both in the mother and in the foetus. Moreover, it is known that the skeletal lesions typical of rickets can seriously worsen the classical pattern of common brachipelvization, resulting in more serious pathologies. Anthropologically, brachipelvization and the evolution to the erect position constitute a peculiarity of our species. Over the centuries, obstetrics has developed complex studies for the evaluation of the pelvis and in particular, for the study of planes, axes and obstetric conjugates. In the nineteenth century and in the first decades of the twentieth century, pelvimetry was carefully practiced in obstetrics to monitor the pathological conditions of the pelvis. The management of postpartum haemorrhage was less theoretically developed and, in obstetric practice, was also represented a frequent cause of maternal death. In the case presented, therefore, obstetric procedures, such as sutures of the cervix of the vagina and the use of the so-called iron perchloride as a haemostatic cauterant, were used to stop bleeding. Ferric chloride is an iron salt (hence it is wrong to call it acid, as it is wrong to call it perchloride). The haemostatic action of the latter has been known for a very long time, but owing to its caustic action, which deeply manifests itself in the tissues, it has been absolutely abandoned in modern obstetric practice. Indeed, the report reveals that ferric chloride gave the tissues inside the uterus a leathery consistency, without - however - managing to save the mother’s life., The typical modern native from Trieste is tall - they are the tallest among Italians -, over 65 years old - Trieste has the highest seniority index in Italy - and loves spending time at the seaside or eating out with friends (no statistic data are available but you can witness that with a day trip to the city!). A group of 41 individuals who lived in the Early Middle Ages in Tergeste (the modern Trieste), seems to have a lot in common with the modern inhabitant of Trieste: they were tall (average Trotter and Gleser stature for male and female are 174.2 cm and 163.2 cm, respectively) and most of them were over 55 yeas old at death (more than 50% of adult males and 50% of adult females). Moreover, some of them spent a significant amount of time in the sea, which can be seen by the auditory exostosis in males and high frequency of third distal tibiae and fibulae periostitis in adult mature females – findings suggestive of a long time spent in the sea water looking for clams. Medieval archaeological layers in Trieste show plenty of shells. These people also loved eating: the four eldest male skeletons show marks of DISH, a pathology clearly associated with metabolic disorders. One of them may have died of suffocation caused by a small-size herbivore distal humerus epiphysis showing clear signs of slicing – a morsel of stew – found on C5/C6. Talking of skeletal remains, this diagnosis can be only a suggestion; nevertheless, dysphagia leading to suffocation in the elderly is a classic. These people were buried out of the walled circle of the ancient Tergeste and not so far from the Madonna del Mare, an early Christian Basilica built in the V/VI century, on the site called Domus Mariae. In 3 out of a total of 21 tombs (the number is low because of the overlapping of different bodies), we find a skull placed in a ritual position close to the left ankle of the body. The following are the combinations found and the age at death in each of these burials: female skeleton aged 30-40/skull of child aged 4; male skeleton aged 50-60/skull of female aged more than 50; male skeleton aged 30-45/skull of male aged more than 50. In the last mentioned tomb, besides the skull to the left of the ankle there are two other skulls: one between the femora, belonging to a man aged about 20-25, and the other one to the left of his skull, belonging to a child aged about 9. The skull connected to the buried skeleton presents a trepanation probably made by scarnification: an oval hole of about 17 mm x 12 mm, on the left parietal bone. The lesion edges clearly demonstrate survival after trepanation, possibly not longer than 1 or 2 months. In fact, in the inner table, near the hole, some coral-like new bone lesions reveal a meningeal involvement. Neither the skull nor the other bones reveal traumatic lesions; we found only a sternal foramen, the lack of fusion of the transverse foramens of C2 and a osteochondritis dissecans of the right capitellum., In the Middle Ages few examples of forensic medicine had appeared, even if imperfect and sporadic, in the field of healthcare art. In the Renaissance they took on greater consistency and framed themselves better in more defined limits. The two reports presented here are part of a trial against Jews in the city of Trento in 1475. This work takes into consideration what happened, the historical period, the trial and the sentences, and the subsequent historical revision. The study of these reports also analyzes the guidelines available at that time., To understand a disease of the past, historians need to distance themselves from today’s epistemological paradigm. The disease must lie within the epistemological limits in which it is explained. In Western medical texts of the past, from Hippocrates and Galen onwards, it was the sign on the patient that led to diagnosis through a deductive process. A debatable diagnosis by our criteria. It follows that the names attributed to diseases bore several meanings, also due to translations over time from one major language to another (Greek, Arabic, Latin) and moreover in relation to the authors describing the disease – physicians, authors of literary texts, chronicles, hagiographies – representatives of an inhomogeneous medical culture. This led to an acknowledgement of the limits of retrospective diagnosis with reference to the lexicon of the textual sources. A significant example regards erysipelas, which today indicates a precise bacterial disease: this term is first found in various points of the Corpus Hippocraticum, including book III of the Epidemics, in a story subsequently commented on by Galen. From the description of the signs the term is linked to different symptoms or diseases, if interpreted with our criteria. Transliterated into Latin, in mediaeval texts the term is placed under the category of the apostemata – exceedingly complex diseases or disturbances – especially after the translation of Avicenna’s Canon. The association with ignis sacer, an expression of Latin origin, in the De Medicina of Cassius Felix (5th C.) led in some cases to a semantic change, borne out by non-medical sources. Ignis sacer in fact, independently of its oldest meaning, first came into use in chronicle sources from the 11th century, to indicate “burning” epidemics, in which ergotism and gangrene in general may be recognised. Renaissance medical sources, following direct translations of Greek medical texts without mediation of the Arabic, tended in part to recover the meaning of the term erysipelas indicated by Galen. In Hoffman’s Dissertatio of 1729, albeit with due precautions, we may recognise the symptoms of present day erysipelas, although the physician’s association with Rosa and with fuoco selvatico begs consideration of the polysemy of two nosographic expressions found in “popular” culture in many areas of Europe. The aim of this presentation is to analyse the polysemy of a nosographic term that has come down to us from the Hippocratic tradition, together with its changes in meaning over time and in accordance with contexts. A paradigmatic case for highlighting the difficulty facing historians wishing to carry out a retrospective diagnosis by means of the medical lexicon., Paolo Gorini (1813-1881) was one of the first scientists who experimentedwith the“petrification” of corpses, a particular technique used to obtain an artificial preservation of bodies, which found wide application in Italy in the 19th century. This technique allowed the exact features of the deceased to be maintained and for tissue, internal organs and hair to be preserved, mostly in a state of stone hardness. This specific mechanism was based on the replacement of biological liquids with chemical preservatives obtained through intravascular injections. Paolo Gorini performed “petrification” on hundreds of specimens, onentire cadavers as well as on parts of corpses, most of which are now housed in the Paolo Gorini Anatomical Collection of Lodi. Lodi is also home to the manuscript with the two formulas used by Gorini to petrify corpses: “a sulfuric acid solution in the proportion of ten percent or an alcohol-saturated solution of mercuric bichloride and muriate of calcium in the proportion that the volume of the first is ten times that of the second”. The aim of our work is to verify the preservation status of skin that was subjected to “petrification” by Gorini. Our study was carried out on an entirely petrified body of an unknown individual held in the aforementioned collectionat Lodi. The man had been affected by a widespread bulbous-bullous infection, possibly smallpox or pellagra. A superficial fragment of skin, free of lesions, was biopsied from the latero-plantar region of the right foot. The analysis was performed using microscopic slides following the inclusion of the samples in epoxy resin, as well as by a stain with hematoxylin-eosin and Masson’s trichrome. Other sections were stained via immunohistochemical technique with anti-cytokeratin antibodies (AE1, AE3) and vimentin. The histological investigations revealed discretely preserved epithelial tissue, with a structure that is still recognizable on the tangential sections. It is possible to distinguishan easily detachable epithelium of the stratum corneum and deeper, more cohesive, layers (stratum granulosum and spinosum) in which the shadows of nuclei are still recognizable. Histochemical investigations revealed positivity for cytokeratins and negativity for vimentin. In contrast to natural or embalmed mummified bodies, historic petrified specimens have never been histologically analyzed. This first study demonstrates that the “petrification” method performed by Gorini guaranteed good skin preservation, allowing its histological, histo-chemical, metachromatic and antigenic characteristics to be maintained., Far from representing a sample of evidence without any current interest, the anatomical and pathological assemblages stored in academic structures are still a valuable scientific and cultural resource for museum collections. In fact, these findings are able to provide, through their precise historical contextualization, important data on epidemiological aspects and medical knowledge over the time. The lack of suitable places to store them and the absence of human and financial resources together with cultural and emotional barriers regarding the death, damn the biological items to the obscurity, despite attempts to enhance them through systematic activities of cataloguing, restoration, conservation and exhibition. The promotion of a newsensitivity towards these collections as well as the developmentof a network system among academic structures may promote the recovery of this biological heritage. These actions could increase the scientific value of the items as well as the the memory of the past and could consign to museums a new role of “places for the scientific reflection and the epistemological revision”. The public exhibition of the biological findings, in accordance with human dignity as well as ethical values, could be a valuable teaching resource towards the knowledge of the human body and also to promote the health awareness. The exposure of healthy organs and pathological ones –in reflecting of unhealthy behaviours and lifestyles or catastrophic natural events – may encourage a critical reflection on the culture of life. At the same time, past human stories, albeit incomplete and fragmentary, may also be an instrument of education in the culture of death and the values of solidarity. In conclusion, we suggest Museums and collections as vectors of new social relations to be shared with the “community of the living”, in order to promote acts of the highest moral value, through awareness campaigns, on the donation of the post-mortem body for study and research purposes., Pathology Museums house ancient specimens obtained during autopsies and generally used for educational purposes. The collections usually consist of dry and wet specimens showing diseases that no longer exist or with their natural course unmodified by therapies. The preservation of the macroscopic features due to the storage fluid has a great historical and paleopathological interest. In recent years, increasing attention is being paid to the study of the wet specimens by modern techniques. Ancient DNA (aDNA) has been investigated in different specimens from natural history museums, but the experience with human material is still limited. The time elapsed between death and fixation, and the chemical composition of fixation and storage fluid may irreversibly damage the DNA, thus routine techniques may result ineffective. We propose a simple and reliable approach to aDNA collection and extraction from museum wet specimens. Ten wet specimens were selected from the Pathology Collection of Turin and submitted to histopathologic re-evaluation. As the chemical composition of the storage fluids is currently unknown, pH value was measured in each specimen. Four cases representative of different classes of pH were submitted to DNA analysis by conservative sampling. Tiny fragments of tissue were frozen at -20°C to obtain sixty 10 mm-thick sections, collected in microtubes containing 1 ml of digestion solution (75 mM NaCl, 10 mM tris, 0.5 mM EDTA, pH 8.0) and 100 ml of proteinase K solution (18 mg/ml). The samples were incubated at 56°C for 48 h and 50 ml fresh of proteinase K solution were added for 72 h. 400 ml of solution were extracted with magnetic beads using a Roche MAGNA PURE COMPACT instrument. DNA quantity and quality were evaluated using the full absorption spectrum (220/340 nm) obtained by the Nanophotometer P 300 spectrophotometer. DNA concentration in ng/ml and absorbance ratio at 260/280 nm were calculated from 4 ml samples. The quality of DNA was also observed by electrophoretic run in 1.3% of agarose gel. In order to verify DNA integrity, short tandem repeat (STR) analysis was performed using the PowerPlex 16 HS system (PROMEGA) employed for personal identification. The cases were originally diagnosed as lymphosarcoma, uterine myosarcoma, esophageal, gastric, and rectal cancers, pancreas tumor, lung cancer, and pleural sarcoma. The range of pH values was comprised between 1.46 and 4.65. The pH value of the specimens submitted to DNA analysis was 2.56, 3.15, 4.45, and 4.65 and the revised diagnoses were necrotic lung carcinoma, uterine leiomyosarcoma, lung metastases from squamous carcinoma of unknown primary, and from uterine leiomyosarcoma. The first two samples gave negative results on both spectrophotometer and electrophoretic runs. The other two showed a low quantity of DNA (6 ng/ml; 7 ng/ml) with an absorbance ratio of 1.53 and 1.50 at the spectrophotometric analysis. The electrophoretic analysis showed a light band of DNA with molecular weight around 1000 bp in both samples. STR analysis displayed DNA fragmentation, evidenced by ladderization of the electropherograms result. The amplification of amelogenin STRs of chromosome X allowed the precise identification of one patient. It is well known that DNA is better preserved in alkaline medium, but its quantity and quality may be acceptable also in specimens preserved at pH around 4.5. Museum wet specimens may represent a valid source of aDNA to investigate genetic molecular features of ancient diseases. The measurement of pH value of the storage fluid may be useful as a screening method for aDNA preservation., Investigating multiple traumas observed in an individual or among members of an entire historic community has always been an area of great interest for paleopathologists and bioarchaeologists. One task faced by paleopathologists is related to the nature of the traumatic event. Such violent events can be accidental or intentional in origin. Intentional violence might be self-inflicted or an act from another person. Once this has been determined, discriminating between multiple injury events and a single event (with multiple fractures) is challenging. While assessing the skeletal collection from the post-classical necropolis of Selvicciola (Viterbo, Latium, Italy; 4th-8th centuries AD), an adult male burial revealed a unique pattern of healed injuries. This male (T 90/5) was buried without grave goods. His tomb is located far from the church, which is the centre of the necropolis. T 90/5 is part of a specific burial group of Longobards situated in South-Eastern funerary area (dated to later period of the 7th century AD). Of these elements we located 6 fractures. This included a well healed nasal fracture, right clavicle fracture, a right scapula fractured with healing along the entire superior body (glenoid fossa to vertebral border), left scapula with an acromion process fracture healed but unfused, a healed rib right fracture and a left femoral neck fracture (unfused). This last facture appears to have happened a few months before his death. The lower edge of the fracture on the femur is well healed with a line of 2 mm of new bone formation. Moreover, the periosteal surface under the neck shows eburnation compatible with the eburnation of the inner part of the head of the femur. The inner surface of the femoral head shows polished remodelled trabeculae lesion. Its creation can likely suggest that a pseudo-articulation between the edge of the diaphysis and the head of the femur was formed as a result of movement of the joint area after the fracture occurred. Also related to the health status of the male is the considerable state of DJD of long bones, as well as the significant evidence of vertebral OA and Schmorl’s nodes. Towards the end of his life this individual was suffering from a number of chronic problems, which produced skeletal lesions specific to biomechanics and old age. Some of this might have been influenced by the trauma experienced earlier in life. Likely, at the end his life his last fracture might have been due to osteoporosis of the femoral neck. The survival of this man testifies to community care and a high value given to human life. The variety of implications in this case-study inform us the care for this individual, that for most part of his life was a disable. Not only the injuries, but also the pain suffered had to affect his daily life. In the end, the protocol of cares was realized by the community at least two times; for the first pattern of trauma (nose, shoulders, clavicle and rib) and for the femur break., Rheumatoid arthritis is a chronic, systemic, inflammatory condition that starts from a synovitis, leading to diffuse erosions in the marginal area of joints and finally conducting to articular deformity and destruction of bone ends. The aetiology of the disease is unknown but multiple genetic and environmental factors have been linked to its development. According to clinical studies, 10-30% of cases undergo healing of lesions and spontaneous remission of the disease. Today’s approach to inflammatory diseases is conditioned by the early diagnosis, thanks to the evolution of the diagnostic methods and by the mitigation of drug therapies. In the past, the remission was entrusted only to the individual’s immune resistance. A case of erosive polyarthropathy has been discovered in an elderly male individual recovered from the medieval cemetery of San Biagio in Cittiglio (northern Italy). The well preserved skeleton was unearthed in the external area in front of the church access and, according to the archaeological stratigraphy, it dates back to a period between the 12th and the 13th century. The bone elements, following macroscopic and microscopic analysis, exhibit several erosive lesions with symmetrical distribution, affecting firstly the appendicular skeleton of the little joints of hands and feet and other larger joints, such as the shoulder, elbow and hip. The bony tissue involved by the erosions is the so-called “bare area”, in the marginal region of the joints, where the synovium membrane-lined bone is found. The diagnosis of this erosive polyarthritis is complicated by the mild expression of the lesions and by the presence of a subtle sclerotic border to some erosions radiographically observed. Next, a careful differential diagnosis was necessary to clarify the aetiology of the polyarthropathy; the skeletal distribution of the lesions and their macroscopic and radiological appearance are suggestive of a case of rheumatoid arthritis-like polyarthropathy. A hypothetical remission phase of the disease, as demonstrated by the frequent presence of smoothed borders and sclerosed margins on radiographic images, is also suggested. Co-existence of diffuse marginal lipping, joint degeneration and severe areas of eburnation, is also recorded, suggesting a co-morbidity of the erosive condition with osteoarthritis, which is compatible with the advanced age of the individual. With this medieval case, we present new evidence of the existence of erosive arthritis and, specifically, of rheumatoid arthritis-like polyarthropathy in Europe before the discovery of the Americas, entering into the long debate about the antiquity of the disease that, firstly, was considered as originating in the New World and subsequently spread to the Old World. On the basis of this and other already published cases, rheumatoid arthritis seems to have been present in Europe more anciently than was previously thought., Between 2012-2014 the Superintendence of Archaeology, Fine Arts and Landscape of Emilia-Romagna, has conducted archaeological fieldworks on a large Late Medieval cemetery (14th-16th century), which archaeological and documentary sources attribute to a Jewish context. The Laboratory of Bioarchaeology and Forensic Osteology of University of Bologna conducted an anthropological study on a sample of 130 individuals. This contribution aims to present four possible cases of venereal treponematoses (TT. 91, 136, 170, 187). Human skeletal remains of graves 91, 136, 170, and 187 were examined to reconstruct the biological profiles and to conduct paleopathological and tomographic analyses, given the presence of lesions on several anatomical districts. Cranial lesions were present on individuals from TT.91 (M, 25-35 years), 170 (M, 25-35 years) and 136 (M, 15-18 years), in which simultaneous destructive and proliferative processes (caries sicca) with focal destruction and remodeling of the external surface and diploe are denoted. Long bones of these three individuals also present osseous alteration such as gummatous osteoperiostitis, with an increased bone density and non-uniform thickening. Individual of T. 187 (11-12 years) presents a hole (3 cm Ø) located on frontal bone, whose margins are remodeled with proliferative processes both on ectocranial and endocranial sides. These lesions are likely linked to treponematosis (bacterial infection by Treponema), interpreted as venereal syphilis. After differential diagnoses, we suppose the individuals of TT. 91 and 170 were likely affected by an advanced stage of the infection, while individual T. 136 seems to have been affected by a tardive congenital form of the disease. Lesions of individual of T.187 suggest an infective origin, but poor bone preservation prevents a clearer interpretation. These cases of treponematosis, possibly linked to venereal syphilis, are relevant for paleoepidemic aspects, as well as contributing to only few Italian osteological cases dating to the same period., Spinal tuberculosis (STB) is a well-known disease in paleopathology. Paleopathologists have highlighted in the last decades some morphological criteria for its diagnosis. Commonly, we are witnessing the destruction of the intervertebral disc space and the adjoining vertebral bodies, the collapse of the vertebrae and the anterior wedging which lead to a structural kyphosis classifiable in gibbus deformity. Here we present the probable STB case of a male subject, 55 years, concealed between the 18th and the first half of the 20th century in the Franciskcan monastery of Azzio, Varese, Italy. The skeleton was found both in an optimal state of preservation and representation. Anthropological analysis was performed according to Buikstra and Ubelaker standards. Paleopathological diagnosis was conducted thanks to macroscopic, microscopic and radiographic analysis, also in order to perform the differential diagnosis. Even if STB was widely present in the last centuries in northwestern Italy, only few paleopathological cases was directly studied., Cystic echinococcosis (CE) is a zoonosis caused by Echinococcus granulosus. The life cycle of the parasite develops in the canids, which house the adult tapeworm in the intestine, and in the intermediate mammal hosts. Humans are occasional dead-end hosts, infected by eggs ingestion via fecal-oral route. The larvae from the digested eggs penetrate the human gut wall and are disseminated throughout the body by the blood. The soft tissues involved at the level of the capillaries may host the larvae, and the hydatid cyst can develop in different organs. The liver is the first organ that the larvae encounter through the blood stream and consequently it is the most frequently involved; it is followed by the lungs and then other organs in frequency. The hydatid cyst is a fluid-filled formation that grows centrifugally and that can survive in the intermediate host for years. In 10 years, it can growth to a diameter of 15-20 cm. Inside hyaline outer membrane, a cellular germinating layer produces microcystic structures that develop scolices. The scolices pouring out of the cyst develop one or more cysts that can reach every tissue. The life cycle is completed when the definitive host feeds on organs of the intermediate host that contain fertile metacestodes. Death of the germinating layer within the metacestode produces calcification of the cyst wall in the intermediate host. Calcified hydatid cysts found as archaeological finds are generally associated with skeletal remains in the thoraco-abdominal site. In archaeological records, the presence of echinococcosis is underestimated, and the find is relatively rare for different reasons: 1) difficulty of recognition by archaeologists; 2) need of accuracy in excavation and recovery of osteoarcheological remains; 3) fragility of calcified formation in the soil. Furthermore, the taphonomic alterations can cause the translation from the original site of the calcified formation and undermine the recognition of the organ affected. In Italy there are only two archaeological samples of calcified formation, most probably of echinococcosis origin, described in the paleopathological literature: one from Siena (13th-14th centuries) and one from Abruzzo (early 20th century). In this report we describe another calcified formation found in the archaeological excavation of the monastic site of Badia Pozzeveri, near Lucca, Tuscany, for which we propose a diagnosis of CE. This finding comes from a privileged lithic coffin built on the northern side of the monastic Church of San Pietro. The grave was used in the 13th century as collective burial by the same laical family group. Calcification, associated with a female individual of about 35-45 years, was discovered in the thoraco-abdominal region. We propose the diagnosis of hydatid cyst from Echinococcus granulosus based on gross morphology, micro-morphology, and a multicomponent approach with cone beam computed tomography, SEM/EDS and stable isotope analysis., Biparietal osteodystrophy (BO), with symmetrical and bilateral thinning of the parietal bones, is a condition rarely discussed in the paleopathological literature. In the past, it has been described as a non-metric trait, anatomical variation and development anomaly. Even though the aetiology is still unknown, today it is described as a pathological condition. In many individuals it appears to be age-related, therefore it has been attributed to osteoporosis, postmenopausal and senile atrophy. Other causes could be congenital and hereditary transmission. Here, we presented a possible case of BO, detected on the remains of a 50-years-old female subject (Tomb 8) excavated in the cemeterial area of the medieval church of Sant’Agostino in Caravate (Varese). Anthropological analysis was performed in accordance with the standards proposed by Buikstra and Ubelaker. Paleopathological conditions were evaluated macroscopically and microscopically. Moreover, CT scan was carried out to investigate pathological evidences. Even if the cause of this uncommon condition is not yet well understood, the present case is highly significant as it enters into the debate upon the aetiology of the disease., Atherosclerosis and its complications represent an important health problem throughout the modern world, although it seems to have accompanied humanity since its beginnings. Important documents have been reported in mummified remains. Atheromas may undergo extensive deposition of calcium and bone metaplasia (Monckeberg’s arteriosclerosis) and persist to the disintegration of the soft tissues. A complete 45-55-years-old male skeleton, 165 cm tall, from bishop’s Palace in Ivrea (Turin) was discovered during archaeological excavation in 2016. Radiocarbon analysis dated the skeleton to 1400-1600 AD. During skeletal preparation in laboratory, an ectopic biological calcification tubular shaped (19 mm in length and 7 mm in diameter) were detected among pelvic bones. At the macroscopic examination, the finding appears as an irregular tubular calcification; in cross section, the mineralized deposits span the entire volume of the lumen and some bony trabeculae in the central space are well defined. Severe calcification of the blood vessel is supposed. Calcification along the expected course of an artery/vessel was considered to be probable atherosclerosis. In relation to the anatomical localization it is supposed to be an atherosclerotic calcification of iliac or femoral artery. The case study reports an uncommon finding of arterial/vessel calcification detected on skeletal remains of an adult male from the post-medieval period. Since the age plays an important role in atherosclerosis, we do not exclude that vascular calcifications affected arteries in many regions of the body. The presented paleopathological specimens suggest that our knowledge of risk factors and the etiology of atherosclerosis are incomplete. A chronic inflammatory burden may have played a greater role than previously considered in ancient cultures and population including upper classes of Italian Renaissance. While increasingly prevalent with age in ancient and modern cultures, a strong gene-environmental interplay is established in the development of atherosclerosis across the lifespan. While genes create the vulnerability, the environment determines when and if atherosclerosis becomes manifest clinically.
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- 2019
3. C42 - The ENDOPREDICT® molecular test for breast cancer prognosis: clinical-pathological correlations and therapeutic implications on a selected cohort of patients
- Author
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Castellano, I., Metovic, J., Balmativola, D., Francia Di Celle, P., Riera, L., Bertetto, O., Mistrangelo, M., Cassoni, P., Marchiò, C., and Sapino, A.
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- 2017
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4. P1.04-45 Immune-Oncology Gene Expression Profiles Allow Lung Cancer Patients’ Stratification and Identification of Responders to Immunotherapy
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Tabbò, F., primary, Annaratone, L., additional, Nocifora, A., additional, Vignale, C., additional, Carnio, S., additional, Metovic, J., additional, Veneziano, F., additional, Scodes, S., additional, Russo, A., additional, Franchina, T., additional, Sini, C., additional, Coco, S., additional, Garlatti, P., additional, Vieri, S., additional, Adamo, V., additional, Boccardo, S., additional, Grossi, F., additional, Cappuzzo, F., additional, Papotti, M., additional, Righi, L., additional, Passiglia, F., additional, and Novello, S., additional
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- 2019
- Full Text
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5. The ENDOPREDICT® molecular test for breast cancer prognosis: clinical-pathological correlations and therapeutic implications on a selected cohort of patients
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Castellano, I., primary, Metovic, J., additional, Balmativola, D., additional, Francia Di Celle, P., additional, Riera, L., additional, Bertetto, O., additional, Mistrangelo, M., additional, Cassoni, P., additional, Marchiò, C., additional, and Sapino, A., additional
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- 2017
- Full Text
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6. Stain normalization in digital pathology: Clinical multi-center evaluation of image quality
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Nicola Michielli, Alessandro Caputo, Manuela Scotto, Alessandro Mogetta, Orazio Antonino Maria Pennisi, Filippo Molinari, Davide Balmativola, Martino Bosco, Alessandro Gambella, Jasna Metovic, Daniele Tota, Laura Carpenito, Paolo Gasparri, Massimo Salvi, Michielli, N., Caputo, A., Scotto, M., Mogetta, A., Pennisi, O. A. M., Molinari, F., Balmativola, D., Bosco, M., Gambella, A., Metovic, J., Tota, D., Carpenito, L., Gasparri, P., and Salvi, M.
- Subjects
Stain normalization ,Image quality ,Qualitative score ,Digital pathology ,H&E staining ,Health Informatics ,Computer Science Applications ,Pathology and Forensic Medicine - Abstract
In digital pathology, the final appearance of digitized images is affected by several factors, resulting in stain color and intensity variation. Stain normalization is an innovative solution to overcome stain variability. However, the validation of color normalization tools has been assessed only from a quantitative perspective, through the computation of similarity metrics between the original and normalized images. To the best of our knowledge, no works investigate the impact of normalization on the pathologist's evaluation. The objective of this paper is to propose a multi-tissue (i.e., breast, colon, liver, lung, and prostate) and multi-center qualitative analysis of a stain normalization tool with the involvement of pathologists with different years of experience. Two qualitative studies were carried out for this purpose: (i) a first study focused on the analysis of the perceived image quality and absence of significant image artifacts after the normalization process; (ii) a second study focused on the clinical score of the normalized image with respect to the original one. The results of the first study prove the high quality of the normalized image with a low impact artifact generation, while the second study demonstrates the superiority of the normalized image with respect to the original one in clinical practice. The normalization process can help both to reduce variability due to tissue staining procedures and facilitate the pathologist in the histological examination. The experimental results obtained in this work are encouraging and can justify the use of a stain normalization tool in clinical routine.
- Published
- 2022
7. Clinical implications of lung neuroendocrine neoplasm classification
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Sergio Harari, Giuseppe Pelosi, Giulia Veronesi, Linda Pattini, Adriana Albini, Jasna Metovic, Angelica Sonzogni, Mauro Papotti, Marco Barella, Metovic, J., Barella, M., Harari, S., Pattini, L., Albini, A., Sonzogni, A., Veronesi, G., Papotti, M., and Pelosi, G.
- Subjects
Pathology ,medicine.medical_specialty ,tumor ,Lung Neoplasms ,diagnosis ,precision medicine ,intratumor heterogeneity ,Carcinoid Tumor ,carcinoma ,lung ,Intratumor heterogeneity ,Carcinoma ,medicine ,Animals ,Humans ,Pharmacology (medical) ,Lung Neuroendocrine Neoplasm ,immuno-oncology ,immunohistochemistry ,marker ,Neuroendocrine ,personalized medicine ,prediction ,prognosis ,Carcinoma, Small Cell ,Lung ,business.industry ,respiratory system ,medicine.disease ,Precision medicine ,digestive system diseases ,respiratory tract diseases ,Carcinoma, Neuroendocrine ,Neuroendocrine Tumors ,medicine.anatomical_structure ,Oncology ,Immunohistochemistry ,Personalized medicine ,business - Abstract
Introduction: Neuroendocrine neoplasms of the lung (Lung NENs) encompass NE tumors (NETs), which are in turn split into typical and atypical carcinoids, and NE carcinomas (NECs), which group together small-cell carcinoma and large-cell NE carcinoma. This classification is the current basis for orienting the daily practice of these patients, with diagnostic, prognostic, and predictive inferences. Areas covered: The clinical implications of lung NEN classification are addressed according to three converging perspectives, which were dissected through an extensive literature overview: (1) how to put intratumor heterogeneity into the context of the current classification; (2) how to contextualize immunohistochemistry markers to improve diagnosis, prognosis, and therapy prediction; and (3) how to use immuno-oncology strategies for life-threatening NECs, which still account for 90% or more of lung NENs. Expert opinion: We provide practical insights to account for intratumor heterogeneity, practice the choice of immunohistochemistry markers, and emphasize once again the added value of immuno-oncology in the setting of personalized medicine of lung NENs.
- Published
- 2021
8. Morphologic and molecular classification of lung neuroendocrine neoplasms
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Fabrizio Bianchi, Jasna Metovic, Fabien Forest, Sergio Harari, Myriam Remmelink, Véronique Hofman, Linda Pattini, Angelica Sonzogni, Mauro Papotti, Lina Carvalho, Paul Hofman, Giuseppe Pelosi, Izidor Kern, Marco Barella, Giulia Veronesi, Metovic, J., Barella, M., Bianchi, F., Hofman, P., Hofman, V., Remmelink, M., Kern, I., Carvalho, L., Pattini, L., Sonzogni, A., Veronesi, G., Harari, S., Forest, F., Papotti, M., and Pelosi, G.
- Subjects
0301 basic medicine ,Pathology ,Lung Neoplasms ,morfološka klasifikacija ,Neuroendocrine tumors ,Signature ,Gene ,0302 clinical medicine ,Molecular classification ,molekularna klasifikacija ,Médecine légale ,Small cell ,Lung ,Tumor ,Stem cell ,molecular classification ,Progression ,General Medicine ,Classification ,Neuroendocrine Tumors ,pljučni tumorji ,medicine.anatomical_structure ,Neuroendocrine ,030220 oncology & carcinogenesis ,Differentiation ,medicine.medical_specialty ,lung neoplasms ,Biology ,Small-cell carcinoma ,Natural history of disease ,Atypical ,Carcinoid ,Carcinoma ,Large cell ,Molecular ,Typical ,Biomarkers, Tumor ,Carcinoma, Neuroendocrine ,Humans ,Pathology and Forensic Medicine ,03 medical and health sciences ,medicine ,Large-cell neuroendocrine carcinoma ,Molecular Biology ,udc:616-006 ,pljučne novotvorbe ,Biologie moléculaire ,Cell Biology ,karcinom ,medicine.disease ,Review and Perspectives ,Pathologie générale ,030104 developmental biology ,morphologic classification ,nevroendokrini tumorji ,lung tumors ,Biologie cellulaire ,neuroendocrine tumors ,Biomarkers - Abstract
Neuroendocrine neoplasms (NENs) of the lung encompass neuroendocrine tumors (NETs) composed of typical (TC) and atypical (AC) carcinoids and full-fledged carcinomas (NECs) inclusive of large cell neuroendocrine carcinoma (LCNEC) and small cell carcinoma (SCLC). NETs and NECs are thought to represent distinct and separate lesions with neither molecular overlap nor common developmental continuum. Two perspectives were addressed regarding the morphologic and molecular classification of lung NENs: (i) a supervised approach by browsing the traditional classification, the relevant gene alterations, and their clinical implications; and (ii) an unsupervised approach, by reappraising neoplasms according to risk factors and natural history of disease to construct an interpretation model relied on biological data. We herein emphasize lights and shadows of the current classification of lung NENs and provide an alternative outlook on these tumors focused on what we currently know about the biological determinants and the natural history of disease., SCOPUS: re.j, info:eu-repo/semantics/published
- Published
- 2021
9. Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases.
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Ferreira da Silva J, Tou CJ, King EM, Eller ML, Rufino-Ramos D, Ma L, Cromwell CR, Metovic J, Benning FMC, Chao LH, Eichler FS, and Kleinstiver BP
- Abstract
Genome editing technologies based on DNA-dependent polymerases (DDPs) could offer several benefits compared with other types of editors to install diverse edits. Here, we develop click editing, a genome writing platform that couples the advantageous properties of DDPs with RNA-programmable nickases to permit the installation of a range of edits, including substitutions, insertions and deletions. Click editors (CEs) leverage the 'click'-like bioconjugation ability of HUH endonucleases with single-stranded DNA substrates to covalently tether 'click DNA' (clkDNA) templates encoding user-specifiable edits at targeted genomic loci. Through iterative optimization of the modular components of CEs and their clkDNAs, we demonstrate the ability to install precise genome edits with minimal indels in diverse immortalized human cell types and primary fibroblasts with precise editing efficiencies of up to ~30%. Editing efficiency can be improved by rapidly screening clkDNA oligonucleotides with various modifications, including repair-evading substitutions. Click editing is a precise and versatile genome editing approach for diverse biological applications., (© 2024. The Author(s), under exclusive licence to Springer Nature America, Inc.)
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- 2024
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10. Gene therapy for the leukodystrophies: From preclinical animal studies to clinical trials.
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Metovic J, Li Y, Gong Y, and Eichler F
- Subjects
- Humans, Animals, Disease Models, Animal, Genetic Therapy methods, Genetic Therapy trends, Clinical Trials as Topic methods
- Abstract
Leukodystrophies are progressive single gene disorders affecting the white matter of the brain. Several gene therapy trials are in progress to address the urgent unmet need for this patient population. We performed a comprehensive literature review of all gene therapy clinical trials listed in www.clinicaltrials.gov through August 2024, and the relevant preclinical studies that enabled clinical translation. Of the approximately 50 leukodystrophies described to date, only eight have existing gene therapy clinical trials: metachromatic leukodystrophy, X-linked adrenoleukodystrophy, globoid cell leukodystrophy, Canavan disease, giant axonal neuropathy, GM2 gangliosidoses, Alexander disease and Pelizaeus-Merzbacher disease. What led to the emergence of gene therapy trials for these specific disorders? What preclinical data or disease context was enabling? For each of these eight disorders, we first describe its pathophysiology and clinical presentation. We discuss the impact of gene therapy delivery route, targeted cell type, delivery modality, dosage, and timing on therapeutic efficacy. We note that use of allogeneic hematopoietic stem cell transplantation in some leukodystrophies allowed for an accelerated path to clinic even in the absence of available animal models. In other leukodystrophies, small and large animal model studies enabled clinical translation of experimental gene therapies. Human clinical trials for the leukodystrophies include ex vivo lentiviral gene delivery, in vivo AAV-mediated gene delivery, and intrathecal antisense oligonucleotide approaches. We outline adverse events associated with each modality focusing specifically on genotoxicity and immunotoxicity. We review monitoring and management of events related to insertional mutagenesis and immune responses. The data presented in this review show that gene therapy, while promising, requires systematic monitoring to account for the precarious disease biology and the adverse events associated with new technology., Competing Interests: Declaration of competing interest F.S. Eichler receives research support from NINDS (R01NS072446, R01NS082331, U54NS115052), institutional contracts from ASPA Therapeutics, Sanofi, Ionis, bluebird bio and Minoryx Therapeutics, and performs personal consulting for Sanofi, ASPA Therapeutics, UpToDate, Leal, Takeda, Atlas Venture, Acadia Pharmaceuticals and SwanBio Therapeutics., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
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- 2024
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11. A neuroligin-2-YAP axis regulates progression of pancreatic intraepithelial neoplasia.
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Middonti E, Astanina E, Vallariello E, Hoza RM, Metovic J, Spadi R, Cristiano C, Papotti M, Allavena P, Novelli F, Parab S, Cappello P, Scarpa A, Lawlor R, Di Maio M, Arese M, and Bussolino F
- Subjects
- Humans, Neuroligins, Cell Transformation, Neoplastic, Pancreatic Neoplasms genetics, Pancreatic Neoplasms pathology, Carcinoma, Pancreatic Ductal genetics, Carcinoma, Pancreatic Ductal pathology, Carcinoma in Situ pathology
- Abstract
Pancreatic ductal adenocarcinoma (PDAC) is a tumor with a dismal prognosis that arises from precursor lesions called pancreatic intraepithelial neoplasias (PanINs). Progression from low- to high-grade PanINs is considered as tumor initiation, and a deeper understanding of this switch is needed. Here, we show that synaptic molecule neuroligin-2 (NLGN2) is expressed by pancreatic exocrine cells and plays a crucial role in the regulation of contact inhibition and epithelial polarity, which characterize the switch from low- to high-grade PanIN. NLGN2 localizes to tight junctions in acinar cells, is diffusely distributed in the cytosol in low-grade PanINs and is lost in high-grade PanINs and in a high percentage of advanced PDACs. Mechanistically, NLGN2 is necessary for the formation of the PALS1/PATJ complex, which in turn induces contact inhibition by reducing YAP function. Our results provide novel insights into NLGN2 functions outside the nervous system and can be used to model PanIN progression., (© 2024. The Author(s).)
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- 2024
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12. ALK inhibitors increase ALK expression and sensitize neuroblastoma cells to ALK.CAR-T cells.
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Bergaggio E, Tai WT, Aroldi A, Mecca C, Landoni E, Nüesch M, Mota I, Metovic J, Molinaro L, Ma L, Alvarado D, Ambrogio C, Voena C, Blasco RB, Li T, Klein D, Irvine DJ, Papotti M, Savoldo B, Dotti G, and Chiarle R
- Subjects
- Humans, Anaplastic Lymphoma Kinase genetics, Anaplastic Lymphoma Kinase metabolism, Protein Kinase Inhibitors pharmacology, Protein Kinase Inhibitors therapeutic use, Antigens, Neoplasm, T-Lymphocytes, Cell Line, Tumor, Neuroblastoma drug therapy, Neuroblastoma genetics, Neuroblastoma metabolism
- Abstract
Selection of the best tumor antigen is critical for the therapeutic success of chimeric antigen receptor (CAR) T cells in hematologic malignancies and solid tumors. The anaplastic lymphoma kinase (ALK) receptor is expressed by most neuroblastomas while virtually absent in most normal tissues. ALK is an oncogenic driver in neuroblastoma and ALK inhibitors show promising clinical activity. Here, we describe the development of ALK.CAR-T cells that show potent efficacy in monotherapy against neuroblastoma with high ALK expression without toxicity. For neuroblastoma with low ALK expression, combination with ALK inhibitors specifically potentiates ALK.CAR-T cells but not GD2.CAR-T cells. Mechanistically, ALK inhibitors impair tumor growth and upregulate the expression of ALK, thereby facilitating the activity of ALK.CAR-T cells against neuroblastoma. Thus, while neither ALK inhibitors nor ALK.CAR-T cells will likely be sufficient as monotherapy in neuroblastoma with low ALK density, their combination specifically enhances therapeutic efficacy., Competing Interests: Declaration of interests D.A. is an employee of Celldex Therapeutics, a company that developed anti-ALK antibodies for therapeutic applications. E.B., W.-T.T., and R.C. filed a patent related to this work covering the development of ALK.CAR-T cells., (Copyright © 2023 Elsevier Inc. All rights reserved.)
- Published
- 2023
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13. Impact of Caloric Restriction in Breast Cancer Patients Treated with Neoadjuvant Chemotherapy: A Prospective Case Control Study.
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Castellano I, Gallo F, Durelli P, Monge T, Fadda M, Metovic J, Cassoni P, Borella F, Raucci C, Menischetti M, Beano A, Migliaretti G, and Finocchiaro C
- Subjects
- Humans, Female, Neoadjuvant Therapy adverse effects, Case-Control Studies, Caloric Restriction, Lymph Nodes, Breast Neoplasms drug therapy, Breast Neoplasms pathology
- Abstract
Background and aims: It is well established that caloric restriction (CR) may influence metabolic and hormonal factors involved in cancer development and progression. Recently, several studies have demonstrated that CR may have a favorable impact on the response to systemic therapy in breast cancer (BC) patients. However, there is a lack of data regarding the influence of CR during neoadjuvant chemotherapy (NACT). Our study's primary aim was to evaluate CR's impact on BC patients undergoing NACT. Secondly, we investigated the nutritional efficacy and safety of this intervention. Methods : We performed a prospective, case-control study in two breast units. A diet group consisting of 39 patients undergoing NACT and CR was enrolled in our study at the same time. CR consisted of a 30% reduction in caloric intake, which increased to 50% on the days before, during, and after the administration of chemotherapy. A control group of 60 patients that underwent the same treatment approach only followed the general dietary recommendations for BC according to WCRF guidelines. The diet group was monitored during the study for both dietary adequacy and weight trends. Results: CR combined with NACT showed a statistically significant therapeutic response in tumor size (OR 2.94, IC 1.07-8.01, p = 0.009) and lymph node status (OR 3.22, IC 1.22-8.56, p = 0.001) compared to NACT alone, even after the adjustment for all biological parameters. Our data also showed the efficacy and safety of this intervention in both anthropometric and biochemical analyses. Conclusions: Patients who adhered to CR showed a better response to NACT, both in the breast and in the axillary lymph nodes, compared to the patients in the control group. Furthermore, the CR diet combined with NACT showed good tolerance and safety.
- Published
- 2023
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14. Clinical and Pathological Features and Gene Expression Profiles of Clinically Aggressive Papillary Thyroid Carcinomas.
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Metovic J, Cabutti F, Osella-Abate S, Orlando G, Tampieri C, Napoli F, Maletta F, Daniele L, Volante M, and Papotti M
- Subjects
- Humans, Middle Aged, Thyroid Cancer, Papillary genetics, Transcriptome, Hedgehog Proteins genetics, Prognosis, Thyroid Neoplasms pathology, Carcinoma, Papillary genetics, Carcinoma, Papillary pathology
- Abstract
Papillary thyroid carcinoma (PTC) is considered an indolent neoplasm but it may demonstrate aggressive behavior. We aimed to identify clinical and pathological characteristics and molecular signatures associated with aggressive forms of PTCs. We selected 43 aggressive PTC cases based on the presence of metastases at the time of diagnosis, the development of distant metastasis during follow-up, and/or biochemical recurrence, and 43 PTC patients that were disease-free upon follow-up, matching them according to age, sex, pT, and pN parameters. Twenty-four pairs (a total of 48 cases) and 6 normal thyroid tissues were studied using targeted mRNA screening of cancer-associated genes employing NanoString nCounter
® technology. In general, aggressive PTCs showed distinctive clinical and morphological features. Among adverse prognostic parameters, the presence of necrosis and an increased mitotic index were associated with shorter disease-free and overall survivals. Other parameters associated with shorter disease-free or overall survivals include a lack of tumor capsule, the presence of vascular invasion, tumor-infiltrating lymphocytes, fibrosclerotic changes, age > 55 years, and a high pTN stage. Various pathways were differentially regulated in non-aggressive as compared to aggressive PTC, including the DNA damage repair, the MAPK, and the RAS pathways. In particular, the hedgehog pathway was differentially de-regulated in aggressive PTC as compared to non-aggressive PTC cases, being WNT10A and GLI3 genes significantly up- and down-regulated in aggressive PTC and GSK3B up-regulated in non-aggressive PTC cases. In conclusion, our study revealed specific molecular signatures and morphological features in aggressive PTC that may be useful to predict more aggressive behavior in a subset of PTC patients. These findings may be useful when developing novel, tailored treatment options for these patients., (© 2023. The Author(s).)- Published
- 2023
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15. Reply to: Neuroendocrine neoplasms of the breast: diagnostic confusion and future perspectives.
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Metovic J, Sapino A, and Castellano I
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- Humans, Female, Breast, Neuroendocrine Tumors diagnosis, Breast Neoplasms diagnosis, Carcinoma, Neuroendocrine diagnosis
- Published
- 2023
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16. Overexpression of INSM1, NOTCH1, NEUROD1, and YAP1 genes is associated with adverse clinical outcome in pediatric neuroblastoma.
- Author
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Metovic J, Napoli F, Osella-Abate S, Bertero L, Tampieri C, Orlando G, Bianchi M, Carli D, Fagioli F, Volante M, and Papotti M
- Subjects
- Child, Humans, Basic Helix-Loop-Helix Transcription Factors genetics, Intracellular Signaling Peptides and Proteins, Membrane Proteins, Receptor, Notch1 genetics, Repressor Proteins genetics, Repressor Proteins metabolism, Retrospective Studies, Transcription Factors genetics, Transcription Factors metabolism, Neuroblastoma genetics, Neuroblastoma pathology
- Abstract
Pediatric neuroblastoma is responsible for approximately 8-10% of pediatric tumors, and it is one of the leading causes of tumor-related deaths in children. Although significant progress has been made in the characterization of neuroblastoma in recent years, the mechanisms influencing the prognosis of neuroblastoma patients remain largely unknown. Our aim was to investigate if the major neuroendocrine-associated transcriptional drivers, including ASCL1, NEUROD1, DLL3, NOTCH1, INSM1, MYCL1, POU2F3 and YAP1 are correlated with specific clinical and pathological characteristics. We selected a retrospective series of 46 primary pediatric neuroblastoma, composed of 30 treatment-naïve and 16 post-chemotherapy cases. Gene expression levels were explored by means of quantitative real-time PCR. An increased expression of NOTCH1 (p = 0.005), NEUROD1 (p = 0.0059), and YAP1 (p = 0.0008) was found in stage IV tumors, while the highest levels of MYCL1 and ASCL1 were seen in stages IVS and III, respectively (p = 0.0182 and p = 0.0134). A higher level of NOTCH1 (p = 0.0079) and YAP1 (p = 0.0026) was found in cases with differentiating morphology, while high mitosis-karyorrhexis index cases demonstrated significantly lower levels of POU2F3 (p = 0.0277). High expression of NOTCH1 (p = 0.008), NEUROD1 (p = 0.026), INSM1 (p = 0.010), and YAP1 (p = 0.005) together with stage IV (p = 0.043) was associated with shorter disease-free survival. In summary, our data indicate that the assessment of gene expression levels of neuroendocrine-lineage transcription factors might help to identify neuroblastoma patients with the risk of relapse., (© 2022. The Author(s).)
- Published
- 2022
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17. Neuroendocrine neoplasms of the breast: diagnostic agreement and impact on outcome.
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Metovic J, Cascardi E, Uccella S, Maragliano R, Querzoli G, Osella-Abate S, Pittaro A, La Rosa S, Bogina G, Cassoni P, Marchiò C, Sapino A, Castellano I, and Papotti M
- Subjects
- Humans, Reproducibility of Results, Cell Differentiation, Neuroendocrine Tumors diagnosis, Neuroendocrine Tumors pathology, Carcinoma, Neuroendocrine diagnosis, Carcinoma, Neuroendocrine pathology, Pancreatic Neoplasms pathology
- Abstract
The classification of breast neuroendocrine neoplasms (Br-NENs) was modified many times over the years and is still a matter of discussion. In the present study, we aimed to evaluate the diagnostic reproducibility and impact on patient outcomes of the most recent WHO 2019 edition of breast tumor classification, namely, for neuroendocrine tumors (NETs) and neuroendocrine carcinomas (NECs). This multicentric observational study included 287 breast neoplasms with NE differentiation. The cases were blindly classified by three independent groups of dedicated breast and/or endocrine pathologists following the 2019 guidelines. Diagnostic concordance and clinical impact were assessed. We observed only a moderate overall diagnostic agreement across the three centers (Cohen's kappa 0.4532) in distinguishing NET from solid papillary carcinomas (SPCs) and no special type carcinomas (NST) with NE differentiation. Br-NENs were diagnosed in 122/287 (42.5%) cases, subclassified as 11 NET G1 (3.8%), 84 NET G2 (29.3%), and 27 NEC (9.4%), the latter group consisting of 26 large-cell and 1 small-cell NECs. The remaining 165/287 (57.5%) cases were labeled as non-NEN, including SPC, mucinous, NST, and mixed NE carcinomas. While NET and non-NEN cases had a comparable outcome, the diagnosis of NECs showed negative impact on disease-free interval compared to NETs and non-NENs (p = 0.0109). In conclusion, the current diagnostic classification of Br-NENs needs further adjustments regarding morphological and immunohistochemical criteria to increase the diagnostic reproducibility among pathologists. Our data suggest that, apart from high-grade small- and large-cell NECs, Br-NENs behave like non-NEN breast carcinomas and should be managed similarly., (© 2022. The Author(s).)
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- 2022
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18. FOXA1 in Breast Cancer: A Luminal Marker with Promising Prognostic and Predictive Impact.
- Author
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Metovic J, Borella F, D'Alonzo M, Biglia N, Mangherini L, Tampieri C, Bertero L, Cassoni P, and Castellano I
- Abstract
The present review focuses on the function of the forkhead protein FOXA1 in breast cancer (BC) in relation to steroid hormone receptors. We explored the currently available analytic approaches for FOXA1 assessment both at gene and protein levels, comparing the differences between the available techniques used for its diagnostic assessment. In addition, we elaborated on data regarding the prognostic and predictive role of this marker in BC based on several studies that evaluated its expression in relation to the outcome and/or response to therapy. FOXA1, similar to the androgen receptor (AR), may have a dual role in BC according to hormonal status. In luminal cancers, its expression contributes to a better prognosis, while in triple-negative breast cancers (TNBC), it implies an adverse outcome. Consequently, we observed that FOXA1-positive expression in a neoadjuvant setting may predict a lack of response in luminal BC as opposed to TNBC, in which FOXA1 allegedly increases its chemosensitivity. In conclusion, considering its accessible and convenient identification by immunohistochemistry, its important impact on prognosis, and its suitability to identify patients with different responses to chemotherapy, we propose that FOXA1 could be tested in routine diagnostics as an additional prognostic and predictive marker in BC.
- Published
- 2022
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19. Stain normalization in digital pathology: Clinical multi-center evaluation of image quality.
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Michielli N, Caputo A, Scotto M, Mogetta A, Pennisi OAM, Molinari F, Balmativola D, Bosco M, Gambella A, Metovic J, Tota D, Carpenito L, Gasparri P, and Salvi M
- Abstract
In digital pathology, the final appearance of digitized images is affected by several factors, resulting in stain color and intensity variation. Stain normalization is an innovative solution to overcome stain variability. However, the validation of color normalization tools has been assessed only from a quantitative perspective, through the computation of similarity metrics between the original and normalized images. To the best of our knowledge, no works investigate the impact of normalization on the pathologist's evaluation. The objective of this paper is to propose a multi-tissue (i.e., breast, colon, liver, lung, and prostate) and multi-center qualitative analysis of a stain normalization tool with the involvement of pathologists with different years of experience. Two qualitative studies were carried out for this purpose: (i) a first study focused on the analysis of the perceived image quality and absence of significant image artifacts after the normalization process; (ii) a second study focused on the clinical score of the normalized image with respect to the original one. The results of the first study prove the high quality of the normalized image with a low impact artifact generation, while the second study demonstrates the superiority of the normalized image with respect to the original one in clinical practice. The normalization process can help both to reduce variability due to tissue staining procedures and facilitate the pathologist in the histological examination. The experimental results obtained in this work are encouraging and can justify the use of a stain normalization tool in clinical routine., Competing Interests: M. Salvi, O.A.M. Pennisi and F. Molinari are equity holders in AEQUIP S.r.l, Turin, Italy. Remaining authors declare no competing interests regarding the publication of this article., (© 2022 The Authors.)
- Published
- 2022
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20. Molecular Subtypes of Extra-pulmonary Neuroendocrine Carcinomas Identified by the Expression of Neuroendocrine Lineage-Specific Transcription Factors.
- Author
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Metovic J, La Salvia A, Rapa I, Napoli F, Birocco N, Pia Bizzi M, Garcia-Carbonero R, Ciuffreda L, Scagliotti G, Papotti M, and Volante M
- Subjects
- Biomarkers, Tumor, Humans, Infant, Newborn, Intracellular Signaling Peptides and Proteins, Membrane Proteins, Repressor Proteins, Transcription Factors, Carcinoma, Neuroendocrine, Lung Neoplasms
- Abstract
Extra-pulmonary neuroendocrine carcinomas (EPNEC) represent a group of rare and heterogenous neoplasms with adverse clinical outcome. Their molecular profile is largely unexplored. Our aim was to investigate if the major transcriptional drivers recently described in high-grade pulmonary neuroendocrine carcinomas characterize distinct molecular and clinical subgroups of EPNEC. Gene expression of ASCL1, NEUROD1, DLL3, NOTCH1, INSM1, MYCL1, POU2F3, and YAP1 was investigated in a series of 54 EPNEC (including 10 cases with mixed components analyzed separately) and in a group of 48 pulmonary large cell neuroendocrine carcinomas (P-LCNEC). Unsupervised hierarchical cluster analysis classified the whole series into four major clusters. P-LCNEC were classified into two major clusters, the first ASCL1/DLL3/INSM1-high and the second (including four EPNEC) ASCL1/DLL3-low but INSM1-high. The remaining EPNEC cases were sub-classified into two other clusters. The first showed INSM1-high and alternative ASCL1/DLL3 or NEUROD1 high expression. The second was characterized mainly by MYCL1 and YAP1 overexpression. In the ten cases with mixed histology, ASCL1, DLL3, INSM1, and NEUROD1 genes were significantly upregulated in the neuroendocrine component. Higher gene-expression levels of NOTCH1 and INSM1 were associated with lower pT stage and negative nodal status. Low INSM1 gene expression was associated with shorter overall survival in the entire case series (p = 0.0017) and with a trend towards significance in EPNEC, only (p = 0.06). In conclusion, our results show that EPNEC possess distinct neuroendocrine-lineage-specific transcriptional profiles; moreover, low INSM1 gene expression represents a novel potential unfavorable prognostic marker in high-grade NECs including those in extra-pulmonary location., (© 2022. The Author(s).)
- Published
- 2022
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21. INSM1 Expression in Breast Neoplasms with Neuroedocrine Features.
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Metovic J, Castellano I, Marinelli E, Osella-Abate S, Sapino A, Cassoni P, and Papotti M
- Subjects
- Aged, Aged, 80 and over, Biomarkers, Tumor metabolism, Breast Neoplasms diagnosis, Breast Neoplasms mortality, Breast Neoplasms pathology, Female, Humans, Immunohistochemistry, Italy epidemiology, Middle Aged, Neuroendocrine Tumors diagnosis, Neuroendocrine Tumors mortality, Neuroendocrine Tumors pathology, Predictive Value of Tests, Prognosis, Retrospective Studies, Sensitivity and Specificity, Survival Analysis, Breast Neoplasms metabolism, Neuroendocrine Tumors metabolism, Repressor Proteins metabolism
- Abstract
According to the 2019 WHO classification of breast tumors, neuroendocrine neoplasms (NENs) are classified into well-differentiated NE tumors (NET) and poorly differentiated NE carcinomas (NEC), while other breast cancers (BCs) of special and no special type with neuroendocrine (NE) features are not incorporated in this scheme anymore. We aimed to assess whether INSM1, a novel NE marker, could have a role in breast NEN subtyping. We selected 63 BCs operated from 2003 to 2018, classified as BCs with NE features, with available clinico-pathological data. Following 2019 WHO criteria, this cohort was reclassified into 37 NETs/NECs, the remaining 26 tumors representing solid-papillary (7), mucinous (7), and mixed type (12) carcinomas with NE differentiation. Chromogranin A (CGA) and synaptophysin (SYN) immunostains were reviewed, and INSM1 was tested by immunohistochemistry. Thirty CGA- and SYN-negative no special type BCs served as negative control. INSM1 was expressed in 52/63 cases of the whole cohort (82.54%). INSM1 positive and negative cases had no significantly different clinico-pathological characteristics. INSM1 expression was not significantly different between the newly reclassified NET/NEC group and other BCs with NE features. No immunoexpression was observed in control BCs. The sensitivity and specificity of INSM1 for the NE phenotype was 82.5% and 100%, respectively, compared to 61.9% and 100% for CGA, and 95.2 and 100% for SYN. In conclusion, INSM1 is as accurate as traditional NE biomarkers to identify NE differentiation in BC. In analogy to standard NE markers, INSM1 could not distinguish NET and NEC from the other BC histotypes with NE differentiation., (© 2021. The Author(s).)
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- 2021
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22. Reply to: Spread Through Air Spaces (STAS): Can an Artifact Really be Excluded?
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Metovic J, Volante M, Righi L, and Papotti M
- Subjects
- Artifacts, Humans, Neoplasm Staging, Adenocarcinoma of Lung pathology, Lung Neoplasms pathology
- Abstract
Competing Interests: Conflicts of Interest and Source of Funding: The authors have disclosed that they have no significant relationships with, or financial interest in, any commercial companies pertaining to this article.
- Published
- 2021
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23. Automated Analysis of Proliferating Cells Spatial Organisation Predicts Prognosis in Lung Neuroendocrine Neoplasms.
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Bulloni M, Sandrini G, Stacchiotti I, Barberis M, Calabrese F, Carvalho L, Fontanini G, Alì G, Fortarezza F, Hofman P, Hofman V, Kern I, Maiorano E, Maragliano R, Marchiori D, Metovic J, Papotti M, Pezzuto F, Pisa E, Remmelink M, Serio G, Marzullo A, Trabucco SMR, Pennella A, De Palma A, Marulli G, Fassina A, Maffeis V, Nesi G, Naheed S, Rea F, Ottensmeier CH, Sessa F, Uccella S, Pelosi G, and Pattini L
- Abstract
Lung neuroendocrine neoplasms (lung NENs) are categorised by morphology, defining a classification sometimes unable to reflect ultimate clinical outcome. Subjectivity and poor reproducibility characterise diagnosis and prognosis assessment of all NENs. Here, we propose a machine learning framework for tumour prognosis assessment based on a quantitative, automated and repeatable evaluation of the spatial distribution of cells immunohistochemically positive for the proliferation marker Ki-67, performed on the entire extent of high-resolution whole slide images. Combining features from the fields of graph theory, fractality analysis, stochastic geometry and information theory, we describe the topology of replicating cells and predict prognosis in a histology-independent way. We demonstrate how our approach outperforms the well-recognised prognostic role of Ki-67 Labelling Index on a multi-centre dataset comprising the most controversial lung NENs. Moreover, we show that our system identifies arrangement patterns in the cells positive for Ki-67 that appear independently of tumour subtyping. Strikingly, the subset of these features whose presence is also independent of the value of the Labelling Index and the density of Ki-67-positive cells prove to be especially relevant in discerning prognostic classes. These findings disclose a possible path for the future of grading and classification of NENs.
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- 2021
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24. Caveolin-1 in Kidney Chronic Antibody-Mediated Rejection: An Integrated Immunohistochemical and Transcriptomic Analysis Based on the Banff Human Organ Transplant (B-HOT) Gene Panel.
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Gambella A, Barreca A, Osella-Abate S, Bottasso E, Giarin MM, Papotti M, Biancone L, Metovic J, Collemi G, Cassoni P, and Bertero L
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Caveolin-1 overexpression has previously been reported as a marker of endothelial injury in kidney chronic antibody-mediated rejection (c-ABMR), but conclusive evidence supporting its use for daily diagnostic practice is missing. This study aims to evaluate if Caveolin-1 can be considered an immunohistochemical surrogate marker of c-ABMR. Caveolin-1 expression was analyzed in a selected series of 22 c-ABMR samples and 11 controls. Caveolin-1 immunohistochemistry proved positive in peritubular and glomerular capillaries of c-ABMR specimens, irrespective of C4d status whereas all controls were negative. Multiplex gene expression profiling in c-ABMR cases confirmed Caveolin-1 overexpression and identified additional genes ( n = 220) and pathways, including MHC Class II antigen presentation and Type II interferon signaling. No differences in terms of gene expression (including Caveolin-1 gene) were observed according to C4d status. Conversely, immune cell signatures showed a NK-cell prevalence in C4d-negative samples compared with a B-cell predominance in C4d-positive cases, a finding confirmed by immunohistochemical assessment. Finally, differentially expressed genes were observed between c-ABMR and controls in pathways associated with Caveolin-1 functions (angiogenesis, cell metabolism and cell-ECM interaction). Based on our findings, Caveolin-1 resulted as a key player in c-ABMR, supporting its role as a marker of this condition irrespective of C4d status.
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- 2021
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25. Recent advances and current controversies in lung neuroendocrine neoplasms ✰ .
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Metovic J, Bianchi F, Rossi G, Barella M, Sonzogni A, Harari S, Papotti M, and Pelosi G
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- Humans, Lung, Carcinoid Tumor therapy, Carcinoma, Large Cell, Carcinoma, Neuroendocrine, Lung Neoplasms genetics, Lung Neoplasms therapy, Neuroendocrine Tumors genetics, Neuroendocrine Tumors therapy
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In the lung, neuroendocrine tumors (NETs), namely typical and atypical carcinoids, and neuroendocrine carcinomas (NECs), grouping small cell carcinoma (SCLC) and large cell neuroendocrine carcinoma (LCNEC), make up for distinct tumor entities according to epidemiological, genetic, pathologic and clinical data. The proper classification is essential in clinical practice for diagnosis, prognosis and therapy purposes. Through an extensive literature survey, three perspectives on lung NENs have been revised: i) criteria and terminology on biopsy or cytology samples of primaries or metastases; ii) carcinoids with elevated mitotic counts and/or Ki-67 proliferation rates; iii) relevance of molecular landscape to identify new tumor entities and therapeutic targets. Furthermore, a dispute about lung NEN development has been raised according to emerging molecular models. We herein provide a pathology update on practical topics in the setting of lung NENs according to the current classification (recent advances). We have also reappraised the development of these tumors by modeling risk factors and natural history of disease (recent controversies). Combining recent advances and controversies may help clarify our biological understanding of lung NENs and give practical information for the clinical decision-making process., (Copyright © 2021 Elsevier Inc. All rights reserved.)
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- 2021
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26. Adrenal Rests in the Uro-genital Tract of an Adult Population.
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Falco EC, Daniele L, Metovic J, Bollito E, De Rosa G, Volante M, and Papotti M
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- Adult, Aged, Aged, 80 and over, Choristoma epidemiology, Female, Humans, Male, Middle Aged, Prevalence, Retrospective Studies, Urogenital Diseases epidemiology, Adrenal Glands, Choristoma pathology, Urogenital Diseases pathology
- Abstract
Ectopic adrenal rests are a rare condition which can be found in various sites, generally in the retroperitoneum or pelvis along the path of gonadal descent. Their real prevalence is unknown. Males are more commonly affected, at least in the pediatric age. Adrenal rests are usually clinically silent and incidentally found in surgical samples, mostly in the pediatric population, and rarely in adults. With the aim of increasing knowledge and estimating the prevalence of ectopic adrenocortical tissue in the adult population, 44 adrenal rests in the urogenital tract of 40 adults are described. These represent approximately 0.07% of the total number of urogenital and gynecological surgeries performed in the 22 considered years. Adrenal rests were identified in the spermatic cord (10 males) and in paraovarian, parasalpingeal, or infundibulopelvic ligament locations (30 females). All but one was incidental findings. One case regarded an adrenocortical carcinoma arisen in adrenal rests. A literature review of adrenal ectopia in the urogenital tract of adults identified 57 reported cases from 53 patients, with similar clinicopathological features as those of our series, with the exception of a lower incidence of parasalpingeal locations. Despite their limited clinical implications, awareness of ectopic adrenal rests is essential also in adults for at least two reasons: (a) to correctly identify sources of adrenocortical hormone production in case of adrenal insufficiency or hormonal imbalance and (b) to avoid misinterpretations in the diagnostic workup of renal cell carcinoma, adrenocortical tumors, and rare gonadal neoplasms, including Sertoli/Leydig cell tumors., (© 2021. The Author(s).)
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- 2021
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27. A Subset of Large Cell Neuroendocrine Carcinomas in the Gastroenteropancreatic Tract May Evolve from Pre-existing Well-Differentiated Neuroendocrine Tumors.
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Pelosi G, Bianchi F, Dama E, Metovic J, Barella M, Sonzogni A, Albini A, Papotti M, Gong Y, and Vijayvergia N
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- Adult, Aged, Aged, 80 and over, Cluster Analysis, DNA Mutational Analysis, Female, Humans, Male, Middle Aged, Carcinoma, Neuroendocrine genetics, Carcinoma, Neuroendocrine pathology, Intestinal Neoplasms genetics, Intestinal Neoplasms pathology, Neuroendocrine Tumors genetics, Neuroendocrine Tumors pathology, Pancreatic Neoplasms genetics, Pancreatic Neoplasms pathology, Stomach Neoplasms genetics, Stomach Neoplasms pathology
- Abstract
In the gastro-entero-pancreatic (GEP) tract, neuroendocrine neoplasms (NENs) include well differentiated neuroendocrine tumors (NETs) and high-grade NE carcinomas (NECs), which are thought to make up separate and mutually exclusive tumor entities. Little is known, however, as to whether there may be any pathogenetic link between them. Clustering analysis of a 10-gene panel generated from a previously reported next-generation sequencing analysis on 48 GEP-NENs with clinical annotations was used in the study. Unsupervised cluster analysis showed three histology-independent clusters, namely, C1, C2, and C3, which accounted for 44% of patients but the entire array of mutations. All but two NECs fell into the clusters, yet with different prevalence rates (p < 0.0001). A model was devised according to which NETs were likely to evolve into NECs upon progression of C3 into C1 and C2, despite different morphology. The median Ki-67 labeling index was 5% in C3 showing better prognosis and 50% in C1 and C2 experiencing worse prognosis, with an impressive intra-tumor heterogeneity of diversely proliferating tumor areas. This study suggests that a subset of large cell NECs in the gastroenteropancreatic tract may evolve from pre-existing well-differentiated NETs., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC part of Springer Nature.)
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- 2021
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28. Amyloid-Rich Pancreatic Neuroendocrine Tumors: a Potential Diagnostic Pitfall in Endoscopic Ultrasound-Guided Fine Needle Aspiration Cytology (EUS-FNAC).
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Gambella A, Falco EC, Metovic J, Maletta F, De Angelis C, Maragliano R, Uccella S, Pacchioni D, and Papotti M
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- Female, Humans, Male, Middle Aged, Retrospective Studies, Amyloid metabolism, Endoscopic Ultrasound-Guided Fine Needle Aspiration methods, Neuroendocrine Tumors diagnosis, Neuroendocrine Tumors pathology, Pancreatic Neoplasms diagnosis, Pancreatic Neoplasms pathology
- Abstract
Pancreatic neuroendocrine tumors (PanNETs) are rare neoplasms that include even rarer variants that may pose different diagnostic problems, especially in fine needle aspiration cytology (FNAC). We describe the diagnostic clues of the amyloid-rich variant of PanNETs in endoscopic ultrasound (EUS)-guided fine needle aspiration cytology (EUS-FNAC). Three cases of PanNETs with an amyloid-rich stromal component were retrieved and retrospectively reviewed. For every case, the pancreatic lesion was investigated by a EUS-FNAC procedure. The final diagnosis was supported by immunocytochemistry and Congo red staining. All cases had similar EUS-FNAC features: neoplastic cells were entrapped in an eosinophilic, homogeneous dense and amorphous matrix. The neuroendocrine nature was confirmed by immunoexpression of synaptophysin and chromogranin A, while the amorphous stroma was characterized as amyloid based on positive Congo red staining. Regarding the hormonal profile, no insulin or proinsulin reactivity was observed, but all cases were diffusely positive for amylin. The diagnosis of uncommon variants of PanNETs, such as the amyloid-rich, is challenging especially in EUS-FNAC procedures because of a unique and misleading morphology, potentially mimicking fibrotic conditions and amyloid deposition within systemic amyloidosis. In cytology specimens, the presence of amorphous material requires amyloid deposition to be considered in the differential diagnosis of pancreatic neoplasms with neuroendocrine phenotype.
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- 2021
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29. Primary lung adenocarcinoma in three adolescent patients affected by bone sarcomas.
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Righi L, Righi A, Vatrano S, Rapa I, Listì A, Metovic J, Rocca M, Salone M, Giovenali P, Sidoni A, Tabbò F, Dei Tos AP, Volante M, and Papotti M
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- Adenocarcinoma complications, Adenocarcinoma pathology, Adenocarcinoma of Lung complications, Adolescent, Bone Neoplasms complications, Bone Neoplasms genetics, Child, Female, Humans, Lung Neoplasms complications, Male, Mutation genetics, Neoplasms, Second Primary, Osteosarcoma complications, Adenocarcinoma of Lung pathology, Bone Neoplasms pathology, Lung Neoplasms pathology, Osteosarcoma pathology
- Abstract
Pediatric primary lung carcinomas are extremely rare. Apart from known associations with congenital adenomatoid malformations, cases of primary lung adenocarcinomas after prolonged treatments of pediatric malignancy have been reported. We describe the morphological and molecular features of three cases of lung adenocarcinoma developed in adolescents aged 8 to 17 years during progression of their bone osteosarcoma or Ewing sarcomas. The morphological features overlapped those of adult lung adenocarcinoma including in situ, minimally invasive, and invasive forms. EGFR gene mutations were found in all three cases by targeted next-generation sequencing. The two patients with Ewing sarcoma had no progression of their lung cancer and no further progression of the metastatic bone tumor after additional chemo- and radio-therapy. Conversely, the osteosarcoma patient refused further treatments after thoracic surgery for metastatic osteosarcoma and locally advanced adenocarcinoma and died 2 years later of widespread distant metastases. Our results indicate that primary lung cancer might originate in pediatric patients during prolonged adjuvant therapies for primary bone neoplasm, and this possibility should be considered in the presence of suspected lung disease progression to correctly monitor the primary tumor evolution and define the appropriate therapeutic strategy at each time point. If appropriately treated, second primary lung cancer may not affect the patients' prognosis. The pathogenetic mechanisms of these rare lung adenocarcinomas are not clear, but the presence of EGFR mutations in all three cases indicates an oncogene addiction of the lung tumor, rather than a direct cancerogenic effect of the sarcoma-related treatment.
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- 2021
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30. Differential Expression Profiles of Cell-to-Matrix-Related Molecules in Adrenal Cortical Tumors: Diagnostic and Prognostic Implications.
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Volante M, Rapa I, Metovic J, Napoli F, Tampieri C, Duregon E, Terzolo M, and Papotti M
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The molecular mechanisms of adrenocortical carcinoma development are incompletely defined. De-regulation of cellular-to-extracellular matrix interactions and angiogenesis appear among mechanisms associated to the malignant phenotype. Our aim was to investigate, employing PCR-based array profiling, 157 molecules involved in cell-to-matrix interactions and angiogenesis in a frozen series of 6 benign and 6 malignant adrenocortical neoplasms, to identify novel pathogenetic markers. In 14 genes, a significant dysregulation was detected in adrenocortical carcinomas as compared to adenomas, most of them being downregulated. Three exceptions-hyaluronan synthase 1 (HAS-1), laminin α3 and osteopontin genes-demonstrated an increased expression in adrenocortical carcinomas of 4.46, 4.23 and 20.32-fold, respectively, and were validated by immunohistochemistry on a series of paraffin-embedded tissues, including 20 adenomas and 73 carcinomas. Osteopontin protein, absent in all adenomas, was expressed in a carcinoma subset (25/73) ( p = 0.0022). Laminin α3 and HAS-1 were mostly expressed in smooth muscle and endothelial cells of the vascular network of both benign and malignant adrenocortical tumors. HAS-1 was also detected in tumor cells, with a more intense pattern in carcinomas. In this group, strong expression was significantly associated with more favorable clinicopathological features. These data demonstrate that cell-to-matrix interactions are specifically altered in adrenocortical carcinoma and identify osteopontin and HAS-1 as novel potential diagnostic and prognostic biomarkers, respectively, in adrenal cortical tumors.
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- 2021
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31. SMARCA2 Deficiency While Preserving SMARCA4 and SMARCB1 in Lung Neuroendocrine Carcinomas.
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Metovic J, Bianchi F, Barella M, Papotti M, and Pelosi G
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- DNA Helicases, Humans, Lung, Nuclear Proteins, SMARCB1 Protein, Transcription Factors genetics, Carcinoma, Neuroendocrine genetics, Lung Neoplasms genetics
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- 2021
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32. Clinical Relevance of Tubular Breast Carcinoma: Large Retrospective Study and Meta-Analysis.
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Metovic J, Bragoni A, Osella-Abate S, Borella F, Benedetto C, Gualano MR, Olivero E, Scaioli G, Siliquini R, Ferrando PM, Bertero L, Sapino A, Cassoni P, and Castellano I
- Abstract
Background: Tubular carcinoma (TC) is a low proliferative grade 1 (G1) breast cancer (BC). Despite its favorable outcome and allegedly lower aggressiveness, patients are treated like other luminal G1 BC, with radiotherapy (RT) and hormonal therapy (HT). We performed: (1) a retrospective study comparing a TC cohort and a control series of luminal G1 BC and (2) a systematic review and meta-analysis focused on TC outcome. Materials and Methods: We selected a series of 572 G1 luminal BC patients [111 TC, 350 not otherwise specified (NOS), and 111 special-type (ST) BC] with follow-up and clinico-pathological data, who underwent local excision followed by RT at Città della Salute e della Scienza Hospital, Turin. Moreover, 22 and 13 studies were included in qualitative and quantitative meta-analysis, respectively. Results: TCs were generally smaller (≤10 mm) ( P < 0.001), with lower lymph node involvement ( P < 0.001). TCs showed no local and/or distant recurrences, while 16 NOS and 2 ST relapsed ( P = 0.036). Kaplan-Meier curves confirmed more favorable TC outcome (DFI: log-rank test P = 0.03). Meta-analysis data, including the results of our study, showed that the pooled DFI rate was 96.4 and 91.8% at 5 and 10 years, respectively. Meta-regression analyses did not show a significant influence of RT nor HT on the DFI at 10 years. Conclusions: Compared to the other G1 BCs, TCs have an excellent outcome. The meta-analysis shows that TC recurrences are infrequent, and HT and RT have limited influence on prognosis. Hence, accurate diagnosis of TC subtype is critical to ensuring a tailored treatment approach., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Metovic, Bragoni, Osella-Abate, Borella, Benedetto, Gualano, Olivero, Scaioli, Siliquini, Ferrando, Bertero, Sapino, Cassoni and Castellano.)
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- 2021
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33. Clinical implications of lung neuroendocrine neoplasm classification.
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Metovic J, Barella M, Harari S, Pattini L, Albini A, Sonzogni A, Veronesi G, Papotti M, and Pelosi G
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- Animals, Carcinoid Tumor classification, Carcinoid Tumor pathology, Carcinoid Tumor therapy, Carcinoma, Neuroendocrine pathology, Carcinoma, Neuroendocrine therapy, Carcinoma, Small Cell classification, Carcinoma, Small Cell pathology, Carcinoma, Small Cell therapy, Humans, Immunohistochemistry, Lung Neoplasms pathology, Lung Neoplasms therapy, Neuroendocrine Tumors pathology, Neuroendocrine Tumors therapy, Precision Medicine, Prognosis, Carcinoma, Neuroendocrine classification, Lung Neoplasms classification, Neuroendocrine Tumors classification
- Abstract
Introduction: Neuroendocrine neoplasms of the lung (Lung NENs) encompass NE tumors (NETs), which are in turn split into typical and atypical carcinoids, and NE carcinomas (NECs), which group together small-cell carcinoma and large-cell NE carcinoma. This classification is the current basis for orienting the daily practice of these patients, with diagnostic, prognostic, and predictive inferences., Areas Covered: The clinical implications of lung NEN classification are addressed according to three converging perspectives, which were dissected through an extensive literature overview: (1) how to put intratumor heterogeneity into the context of the current classification; (2) how to contextualize immunohistochemistry markers to improve diagnosis, prognosis, and therapy prediction; and (3) how to use immuno-oncology strategies for life-threatening NECs, which still account for 90% or more of lung NENs., Expert Opinion: We provide practical insights to account for intratumor heterogeneity, practice the choice of immunohistochemistry markers, and emphasize once again the added value of immuno-oncology in the setting of personalized medicine of lung NENs.
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- 2021
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34. The lobular neoplasia enigma: management and prognosis in a long follow-up case series.
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Metovic J, Abate SO, Borella F, Vissio E, Bertero L, Mariscotti G, Durando M, Senetta R, Ala A, Benedetto C, Sapino A, Cassoni P, and Castellano I
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- Follow-Up Studies, Humans, Hyperplasia, Middle Aged, Prognosis, Breast Neoplasms diagnostic imaging, Breast Neoplasms epidemiology, Breast Neoplasms surgery, Carcinoma in Situ, Carcinoma, Lobular diagnostic imaging, Carcinoma, Lobular surgery
- Abstract
Background: Many oncologists debate if lobular neoplasia (LN) is a risk factor or an obligatory precursor of more aggressive disease. This study has three aims: (i) describe the different treatment options (surgical resection vs observation), (ii) investigate the upgrade rate in surgically treated patients, and (iii) evaluate the long-term occurrences of aggressive disease in both operated and unoperated patients., Methods: A series of 122 patients with LN bioptic diagnosis and follow-up information were selected. Clinical, radiological, and pathological data were collected from medical charts. At definitive histology, either invasive or ductal carcinoma in situ was considered upgraded lesions., Results: Atypical lobular hyperplasia (ALH), lobular carcinoma in situ (LCIS), and high-grade LN (HG-LN) were diagnosed in 44, 63, and 15 patients, respectively. The median follow-up was 9.5 years. Ninety-nine patients were surgically treated, while 23 underwent clinical-radiological follow-up. An upgrade was observed in 28/99 (28.3%). Age ≥ 54 years (OR 4.01, CI 1.42-11.29, p = 0.009), Breast Imaging-Reporting and Data System (BI-RADS) categories 4-5 (OR 3.76, CI 1.37-10.1, p = 0.010), and preoperatory HG-LN diagnosis (OR 8.76, 1.82-42.27, p = 0.007) were related to upgraded/aggressive disease. During follow-up, 8 patients developed an ipsilateral malignant lesion, four of whom were not initially operated (4/23, 17%)., Conclusions: BI-RADS categories 4-5, HG-LN diagnosis, and age ≥ 54 years were features associated with an upgrade at definitive surgery. Moreover, 17% of unoperated cases developed an aggressive disease, emphasizing that LN patients need close surveillance due to the long-term risk of breast cancer.
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- 2021
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35. Diagnostic Value of Conventional PET Parameters and Radiomic Features Extracted from 18F-FDG-PET/CT for Histologic Subtype Classification and Characterization of Lung Neuroendocrine Neoplasms.
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Thuillier P, Liberini V, Rampado O, Gallio E, De Santi B, Ceci F, Metovic J, Papotti M, Volante M, Molinari F, and Deandreis D
- Abstract
Aim: To evaluate if conventional Positron emission tomography (PET) parameters and radiomic features (RFs) extracted by 18F-FDG-PET/CT can differentiate among different histological subtypes of lung neuroendocrine neoplasms (Lu-NENs)., Methods: Forty-four naïve-treatment patients on whom 18F-FDG-PET/CT was performed for histologically confirmed Lu-NEN (n = 46) were retrospectively included. Manual segmentation was performed by two operators allowing for extraction of four conventional PET parameters (SUVmax, SUVmean, metabolic tumor volume (MTV), and total lesion glycolysis (TLG)) and 41 RFs. Lu-NENs were classified into two groups: lung neuroendocrine tumors (Lu-NETs) vs. lung neuroendocrine carcinomas (Lu-NECs). Lu-NETs were classified according to histological subtypes (typical (TC)/atypical carcinoid (AC)), Ki67-level, and TNM staging. The least absolute shrink age and selection operator (LASSO) method was used to select the most predictive RFs for classification and Pearson correlation analysis was performed between conventional PET parameters and selected RFs., Results: PET parameters, in particular, SUVmax (area under the curve (AUC) = 0.91; cut-off = 5.16) were higher in Lu-NECs vs. Lu-NETs ( p < 0.001). Among RFs, HISTO_Entropy_log10 was the most predictive (AUC = 0.90), but correlated with SUVmax/SUVmean (r = 0.95/r = 0.94, respectively). No statistical differences were found between conventional PET parameters and RFs ( p > 0.05) and TC vs. AC classification. Conventional PET parameters were correlated with N+ status in Lu-NETs., Conclusion: In our study, conventional PET parameters were able to distinguish Lu-NECs from Lu-NETs, but not TC from AC. RFs did not provide additional information.
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- 2021
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36. Genomics of High-Grade Neuroendocrine Neoplasms: Well-Differentiated Neuroendocrine Tumor with High-Grade Features (G3 NET) and Neuroendocrine Carcinomas (NEC) of Various Anatomic Sites.
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Uccella S, La Rosa S, Metovic J, Marchiori D, Scoazec JY, Volante M, Mete O, and Papotti M
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- Humans, Neoplasm Grading, Pathology, Molecular, Carcinoma, Neuroendocrine genetics, Carcinoma, Neuroendocrine pathology, Genomics, Neuroendocrine Tumors genetics, Neuroendocrine Tumors pathology
- Abstract
High-grade neuroendocrine neoplasms (HG-NENs) are clinically aggressive diseases, the classification of which has recently been redefined. They now include both poorly differentiated NENs (neuroendocrine carcinoma, NECs) and high proliferating well-differentiated NENs (called grade 3 neuroendocrine tumors, G3 NETs, in the digestive system). In the last decade, the "molecular revolution" that has affected all fields of medical oncology has also shed light in the understanding of HG NENs heterogeneity and has provided new diagnostic and therapeutic tools, useful in the management of these malignancies. Considering the kaleidoscopic aspects of HG NENs in various anatomical sites, this review systematically addresses the genomic landscape of such neoplasm throughout the more common thoracic and digestive locations, as well as it will consider other rare but not exceptional primary sites, including the skin, the head and neck, and the urogenital system. The revision of the available literature will then be oriented to understand the translational relevance of molecular data, by analyzing conceptual issues, clinicopathological correlations, and unmet needs in this field.
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- 2021
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37. Gross Specimen Handling Procedures Do Not Impact the Occurrence of Spread Through Air Spaces (STAS) in Lung Cancer.
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Metovic J, Falco EC, Vissio E, Santoro F, Delsedime L, Massa F, Pittaro A, Osella-Abate S, Cassoni P, Volante M, Righi L, and Papotti M
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- Humans, Neoplasm Invasiveness pathology, Artifacts, Lung Neoplasms pathology, Specimen Handling adverse effects
- Abstract
Spread Through Air Spaces (STAS) is a form of invasion characterized by neoplastic cell dissemination in the lung parenchyma surrounding the outer edge of the tumor. Its possible artifactual origin is widely debated in the literature. The aim of this study is to investigate the potential impact of gross sampling procedures in causing STAS. A prospective series of 51 surgical lung specimens was collected (35 adenocarcinomas, 68.6%; 13 squamous cell carcinomas, 25.5%; 2 large-cell neuroendocrine carcinomas, 3.9%; 1 atypical carcinoid, 2%). The fresh tissue was sectioned with a new and clean blade for each cut, to obtain a tissue slice comprising the upper lung parenchyma, the tumor, and the lower parenchyma. This slice was cut in half and separately processed. The same procedure was repeated in the residual (specular) specimen after formalin fixation. STAS was identified in 33/51 (64.7%) cases, the predominant pattern being cluster formation (29 cases, 87.9%), the remaining 4 cases having single-cell invasion. Comparing STAS detection in upper and lower lung parenchyma areas (ie, before and after the blade crossed the tumor), no significant preferential STAS distribution was observed, indeed being almost overlapping (60.6% and 63.6% for fresh and 61.3% and 65.6% for fixed tissues, respectively). There was no difference between STAS occurrence in freshly cut and fixed corresponding samples. These findings indicate that STAS is not a pathologist-related artifactual event because of knife transportation of tumor cells during gross specimen handling and support the notion that it is a phenomenon preexisting to surgical tissue processing., Competing Interests: Conflicts of Interest and Source of Funding: The authors have disclosed that they have no significant relationships with, or financial interest in, any commercial companies pertaining to this article., (Copyright © 2020 Wolters Kluwer Health, Inc. All rights reserved.)
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- 2021
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38. Proposal of a Panel of Genes Identified by miRNA Profiling as Candidate Prognostic Biomarkers in Lung Carcinoids.
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Rapa I, Votta A, Giorcelli J, Izzo S, Rigutto A, Metovic J, Napoli F, and Volante M
- Subjects
- Activating Transcription Factor 2 genetics, Activin Receptors, Type II genetics, Carcinoid Tumor metabolism, Carcinoid Tumor mortality, Carcinoid Tumor pathology, Disease-Free Survival, Female, Humans, Kruppel-Like Transcription Factors genetics, LIM-Homeodomain Proteins genetics, Lung Neoplasms metabolism, Lung Neoplasms mortality, Lung Neoplasms pathology, Male, Middle Aged, Prognosis, Retrospective Studies, Transcription Factors genetics, Biomarkers, Tumor genetics, Carcinoid Tumor genetics, Gene Expression Profiling, Lung Neoplasms genetics, MicroRNAs metabolism
- Abstract
Aim: To validate the prognostic role of a panel of genes previously uncovered by our group to be specific targets of miRNAs differentially expressed in lung carcinoids with aggressive pathological features., Methods: Four genes, namely, cyclic AMP response element binding protein-1 (CREBP1), activin A receptor type 2B (ACVR2B), LIM homeobox 2 (LHX2), and Krüppel-like factor 12 (KLF12), were identified in a previous study by our group using in silico analysis to be regulated by 3 miRNAs (miR-409-3p, miR-409-5p, and miR-431-5p) that were shown to be downregulated in aggressive lung carcinoids. These genes were analyzed using real-time PCR in a cohort of 102 lung carcinoids. Fifty high-grade lung carcinomas served as control group. Their expression was correlated with the expression of miR-409-3p, miR-409-5p, and miR-431-5p and with clinical pathological parameters and disease-free survival., Results: The expression of all but CREBP1 gene was significantly different between lung carcinoids and high-grade neuroendocrine carcinomas. ACVR2B and LHX2 were significantly inversely correlated with miR-409-3p and miR-409-5p. High levels of ACVR2B and LHX2 were significantly associated with atypical histotype, high tumor grade, and higher proliferation Ki-67 index (all p < 0.05). Low levels of KLF12 were significantly associated with the presence of necrosis and positive nodal status (all p < 0.05). Finally, low KLF12 expression was associated with shorter disease-free survival in lung carcinoids as a whole and in atypical carcinoids, only (all p < 0.001)., Conclusions: ACVR2B, LHX2, and KFL12 are novel potential biomarkers associated with aggressive features in lung carcinoids., (2020 S. Karger AG, Basel.)
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- 2021
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39. Malignant struma ovarii: next-generation sequencing of six cases revealed Nras, Braf, and Jak3 mutations.
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Poli R, Scatolini M, Grosso E, Maletta F, Gallo M, Liscia D, Nelva A, Cesario F, Forte G, Metovic J, Volante M, Arvat E, and Papotti M
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- Female, GTP Phosphohydrolases, High-Throughput Nucleotide Sequencing, Humans, Janus Kinase 3, Membrane Proteins, Mutation, Proto-Oncogene Proteins B-raf genetics, Ovarian Neoplasms genetics, Struma Ovarii genetics, Thyroid Neoplasms
- Abstract
Purpose: Struma ovarii (SO) is a highly specialized ovarian teratoma, consisting of thyroid tissue. Rarely, carcinomas histologically identical to their thyroid counterparts may occur, and are comprehensively defined as malignant struma ovarii (MSO). Their optimal management is controversial, and the molecular profile of the malignant counterpart in the ovary is incompletely known. In this study, the clinicopathological and molecular features of six MSO from different Italian Institutions were analysed, to explore genetic profiles of potential therapeutic interest., Methods: The histopathological features and immunoprofile (according to the known markers Galectin-3, HBME1, cytokeratin 19 and CD56) were reviewed. In addition, all cases underwent genetic analysis with a next-generation sequencing (NGS) hot spot cancer panel detecting mutations in 50 genes involved in cancerogenesis. RET/PTC rearrangements and TERT promoter alterations were also evaluated., Results: Papillary carcinoma in all similar to its thyroid counterpart was found in five of six cases, including classical (two tumors) and follicular variant (three tumors) types. The last case was a poorly differentiated carcinoma. An activating gene mutation, was detected in five of six cases, including two NRAS, two BRAF, and one JAK3 oncogene mutations. No alterations were found in the other panel genes, nor in TERT promoter, or in RET chromosomal regions., Conclusions: MSO is a rare condition. Papillary carcinoma is the predominant malignant type, sharing both histomorphological and molecular features of its thyroid counterpart. Interestingly, the single case of poorly differentiated carcinoma displayed a JAK3 mutation. The presence of such driving mutation could be of potential interest in guiding postoperative treatment.
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- 2021
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40. Morphologic and molecular classification of lung neuroendocrine neoplasms.
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Metovic J, Barella M, Bianchi F, Hofman P, Hofman V, Remmelink M, Kern I, Carvalho L, Pattini L, Sonzogni A, Veronesi G, Harari S, Forest F, Papotti M, and Pelosi G
- Subjects
- Biomarkers, Tumor genetics, Biomarkers, Tumor metabolism, Carcinoma, Neuroendocrine genetics, Carcinoma, Neuroendocrine metabolism, Humans, Lung metabolism, Lung Neoplasms genetics, Lung Neoplasms metabolism, Neuroendocrine Tumors genetics, Neuroendocrine Tumors metabolism, Carcinoma, Neuroendocrine pathology, Lung pathology, Lung Neoplasms pathology, Neuroendocrine Tumors pathology
- Abstract
Neuroendocrine neoplasms (NENs) of the lung encompass neuroendocrine tumors (NETs) composed of typical (TC) and atypical (AC) carcinoids and full-fledged carcinomas (NECs) inclusive of large cell neuroendocrine carcinoma (LCNEC) and small cell carcinoma (SCLC). NETs and NECs are thought to represent distinct and separate lesions with neither molecular overlap nor common developmental continuum. Two perspectives were addressed regarding the morphologic and molecular classification of lung NENs: (i) a supervised approach by browsing the traditional classification, the relevant gene alterations, and their clinical implications; and (ii) an unsupervised approach, by reappraising neoplasms according to risk factors and natural history of disease to construct an interpretation model relied on biological data. We herein emphasize lights and shadows of the current classification of lung NENs and provide an alternative outlook on these tumors focused on what we currently know about the biological determinants and the natural history of disease.
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- 2021
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41. The Oncocytic Variant of Poorly Differentiated Thyroid Carcinoma Shows a Specific Immune-Related Gene Expression Profile.
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Metovic J, Vignale C, Annaratone L, Osella-Abate S, Maletta F, Rapa I, Cabutti F, Patriarca S, Gallo M, Nikiforov YE, Volante M, and Papotti M
- Subjects
- Adenocarcinoma, Follicular immunology, Adenocarcinoma, Follicular mortality, Adenocarcinoma, Follicular pathology, Adenoma, Oxyphilic immunology, Adenoma, Oxyphilic mortality, Adenoma, Oxyphilic pathology, Adult, Aged, Aged, 80 and over, Female, Gene Expression Regulation, Neoplastic, Humans, Male, Microarray Analysis, Middle Aged, Oxyphil Cells pathology, Retrospective Studies, Thyroid Neoplasms immunology, Thyroid Neoplasms mortality, Thyroid Neoplasms pathology, Transcriptome, Tumor Escape genetics, Adenocarcinoma, Follicular genetics, Adenoma, Oxyphilic genetics, Immunity genetics, Oxyphil Cells metabolism, Thyroid Neoplasms genetics
- Abstract
Background: Poorly differentiated thyroid cancer (PDTC) is a rare, follicular cell-derived neoplasm with an unfavorable prognosis. The oncocytic variant of PDTC may be associated with even more adverse outcome than classical PDTC cases, but its specific molecular features are largely unknown. Our aim was to explore the immune-related gene expression profile of oncocytic and classical PDTC, in correlation with clinical and pathological characteristics (including programmed death ligand 1 [PD-L1] expression) and outcome, and in comparison with a control group of well-differentiated follicular carcinomas (WDFCs), including conventional follicular carcinomas (FTCs) and Hürthle cell carcinomas (HCCs)., Methods: A retrospective series of 48 PDTCs and 24 WDFCs was analyzed by means of NanoString technology employing the nCounter PanCancer Immune Profiling panel. Gene expression data were validated using quantitative real-time polymerase chain reaction., Results: Oncocytic PDTCs showed a specific immune-related gene expression profile, with higher expression of LAIR2, CD274, DEFB1, IRAK1, CAMP, LCN2, LY96, and APOE, and lower expression of NOD1, as compared to conventional PDTCs. This molecular signature was associated with increased intratumoral lymphocytic infiltration, PD-L1 expression, and adverse outcome. Three of these genes, CD274, DEFB1, and IRAK1, as well as PD-L1 expression, were also the hallmarks of HCCs as compared to FTCs. By contrast, the panel of genes differentially regulated in PDTCs as compared to WDFCs was unrelated to the oncocytic phenotype., Conclusions: Our results revealed a distinctive immune-related gene expression profile of oncocytic PDTC and confirmed a more aggressive outcome in this cancer subtype. These findings may provide guidance when exploring novel immunotherapeutic options for oncocytic PDTC patients., (© Endocrine Society 2020. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
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- 2020
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42. Correction to: Malignant struma ovarii: next-generation sequencing of six cases revealed Nras, Braf, and Jak3 mutations.
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Poli R, Scatolini M, Grosso E, Maletta F, Gallo M, Liscia D, Nelva A, Cesario F, Forte G, Metovic J, Volante M, Arvat E, and Papotti M
- Abstract
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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- 2020
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43. Noninvasive Follicular Thyroid Neoplasm with Papillary-Like Nuclear Features: From Echography to Genetic Profile.
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Maletta F, Falco EC, Gambella A, Metovic J, and Papotti M
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- Cell Nucleus pathology, Female, Geography, Humans, Middle Aged, Neoplasm Staging, Preoperative Period, Retrospective Studies, Thyroid Gland pathology, Treatment Outcome, Adenocarcinoma, Follicular diagnostic imaging, Adenocarcinoma, Follicular genetics, Genetic Profile, Thyroid Cancer, Papillary diagnostic imaging, Thyroid Cancer, Papillary genetics, Thyroid Neoplasms diagnostic imaging, Thyroid Neoplasms genetics, Ultrasonography methods
- Abstract
In thyroid pathology, the great variety of types and the wide range of aggressiveness of thyroid cancers complicate both diagnosis and management. In 2016, a subset of noninvasive encapsulated follicular variant of papillary thyroid carcinoma was reclassified as noninvasive follicular thyroid tumor with papillary-like nuclear features (NIFTP) to reduce overtreatment of this low-risk tumor that follows a benign course after surgery. Starting from a paradigmatic clinical case, in this short review, we will summarize the ultrasonography, cytological, histological and molecular features of this new entity. In the preoperative settings, the recognition of some peculiar elements may only suggest the possibility of a NIFTP, thus favoring a less aggressive surgical approach. However, the diagnosis of NIFTP can only be made after complete resection of the lesion by detecting well-defined inclusion and exclusion histopathological criteria. Since NIFTP is not 'malignant,' surgery may be considered curative with no further treatment or surveillance needed. NIFTP-related issues, including nodule size, multifocality, oncocytic changes, heterogeneous incidence across different geographical areas and its occurrence in the pediatric age, will be discussed.
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- 2020
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44. Prognostic role of PD-L1 and immune-related gene expression profiles in giant cell tumors of bone.
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Metovic J, Annaratone L, Linari A, Osella-Abate S, Musuraca C, Veneziano F, Vignale C, Bertero L, Cassoni P, Ratto N, Comandone A, Grignani G, Piana R, and Papotti M
- Subjects
- Adolescent, Adult, Aged, Biomarkers, Tumor genetics, Bone Neoplasms pathology, Bone and Bones pathology, Down-Regulation genetics, Female, Giant Cell Tumors pathology, Humans, Immune Tolerance genetics, Male, Middle Aged, Neoplasm Recurrence, Local genetics, Neoplasm Recurrence, Local pathology, Prognosis, Up-Regulation genetics, Young Adult, B7-H1 Antigen genetics, Bone Neoplasms genetics, Bone Neoplasms immunology, Giant Cell Tumors genetics, Giant Cell Tumors immunology, Transcriptome immunology
- Abstract
Giant cell tumor of bone (GCTB) is a locally aggressive and rarely metastatic tumor, with a relatively unpredictable clinical course. A retrospective series of 46 GCTB and a control group of 24 aneurysmal bone cysts (ABC) were selected with the aim of investigating the PD-L1 expression levels and immune-related gene expression profile, in correlation with clinicopathological features. PD-L1 and Ki67 were immunohistochemically tested in each case. Furthermore, comprehensive molecular analyses were carried out using NanoString technology and nCounter PanCancer Immune Profiling Panel, and the gene expression results were correlated with clinicopathological characteristics. PD-L1 expression was observed in 13/46 (28.3%) GCTB (and in 1/24, 4.2%, control ABC, only) and associated with a shorter disease free interval according to univariate analysis. Moreover, in PD-L1-positive lesions, three genes (CD27, CD6 and IL10) were significantly upregulated (p < 0.01), while two were downregulated (LCK and TLR8, showing borderline significance, p = 0.06). Interestingly, these genes can be related to maturation and immune tolerance of bone tissue microenvironment, suggesting a more immature/anergic phenotype of giant cell tumors. Our findings suggest that PD-L1 immunoreactivity may help to select GCTB patients with a higher risk of recurrence who could potentially benefit from immune checkpoint blockade.
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- 2020
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45. Fully automated quantitative assessment of hepatic steatosis in liver transplants.
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Salvi M, Molinaro L, Metovic J, Patrono D, Romagnoli R, Papotti M, and Molinari F
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- Algorithms, Humans, Image Processing, Computer-Assisted, Neural Networks, Computer, Fatty Liver diagnostic imaging, Liver Transplantation
- Abstract
Background: The presence of macro- and microvesicular steatosis is one of the major risk factors for liver transplantation. An accurate assessment of the steatosis percentage is crucial for determining liver graft transplantability, which is currently based on the pathologists' visual evaluations on liver histology specimens., Method: The aim of this study was to develop and validate a fully automated algorithm, called HEPASS (HEPatic Adaptive Steatosis Segmentation), for both micro- and macro-steatosis detection in digital liver histological images. The proposed method employs a hybrid deep learning framework, combining the accuracy of an adaptive threshold with the semantic segmentation of a deep convolutional neural network. Starting from all white regions, the HEPASS algorithm was able to detect lipid droplets and classify them into micro- or macrosteatosis., Results: The proposed method was developed and tested on 385 hematoxylin and eosin (H&E) stained images coming from 77 liver donors. Automated results were compared with manual annotations and nine state-of-the-art techniques designed for steatosis segmentation. In the TEST set, the algorithm was characterized by 97.27% accuracy in steatosis quantification (average error 1.07%, maximum average error 5.62%) and outperformed all the compared methods., Conclusions: To the best of our knowledge, the proposed algorithm is the first fully automated algorithm for the assessment of both micro- and macrosteatosis in H&E stained liver tissue images. Being very fast (average computational time 0.72 s), this algorithm paves the way for automated, quantitative and real-time liver graft assessments., (Copyright © 2020 Elsevier Ltd. All rights reserved.)
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- 2020
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46. Integration of Ki-67 index into AJCC 2018 staging provides additional prognostic information in breast tumours candidate for genomic profiling.
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Vissio E, Metovic J, Osella-Abate S, Bertero L, Migliaretti G, Borella F, Benedetto C, Sapino A, Cassoni P, and Castellano I
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- Breast Neoplasms mortality, Breast Neoplasms pathology, Breast Neoplasms therapy, Cell Proliferation genetics, Female, Genomics, Humans, Immunohistochemistry, Molecular Diagnostic Techniques, Neoplasm Staging, Prognosis, Proportional Hazards Models, Receptor, ErbB-2 metabolism, Receptors, Estrogen metabolism, Receptors, Progesterone metabolism, Retrospective Studies, Breast Neoplasms metabolism, Ki-67 Antigen metabolism
- Abstract
Background: The Eighth edition of the American Joint Committee on Cancer (AJCC) staging system (2018) for breast cancer (BC) introduced the prognostic stage. Moreover, multigene assessment has been indicated to tailor staging in T1/T2/N0, ER-positive/HER2-negative BC. However, many National Health Systems do not provide reimbursement for routine testing. The aim of this study was to assess whether Ki67 proliferation index is prognostically relevant for patients' candidacy for molecular testing., Methods: A retrospective series of 686 ER+/HER2- BC were reclassified using AJCC 2018, and in the group of 521 patients for which AJCC 2018 recommends molecular evaluation, we assessed the prognostic efficacy of a prognostic stage enriched by Ki67 (Ki67-PS), considering Ki67 <20% an alternative to recurrence score <11 provided by Oncotype DX., Results: We found that a group of BCs (35.6%, 58/163) assigned to IB stage by prognostic score were down classified to IA with Ki67-PS. The outcome of these 58 cases overlapped with that of lesions classified as stage IA using prognostic stage, showing a significantly better prognosis compared to IB tumours (HR = 2.79, p = 0.003)., Conclusions: These data suggest that Ki67 may be a reliable marker to enrich the 2018 AJCC prognostic score in BC patients' candidacy for genomic profiling.
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- 2020
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47. Role of Immunocytochemistry in the Cytological Diagnosis of Pulmonary Tumors.
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Metovic J, Righi L, Delsedime L, Volante M, and Papotti M
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- Biopsy, Fine-Needle methods, Bronchoalveolar Lavage methods, Bronchoscopy methods, Humans, Lung pathology, Lung Neoplasms diagnosis, Lung Neoplasms pathology, Sputum metabolism, Biomarkers, Tumor metabolism, Cytodiagnosis methods, Immunohistochemistry methods, Lung metabolism, Lung Neoplasms metabolism
- Abstract
Pulmonary cytology is a challenging diagnostic tool, and it is usually evaluated considering medical history and radiological findings in order to reach an accurate diagnosis. Since the majority of lung cancer patients have an advanced stage at diagnosis, a cytological specimen is frequently the only material available for diagnosis and further prognostic/predictive marker determination. Several types of specimens can be obtained from the respiratory system (including sputum, bronchoalveolar lavage, bronchial brushing, fine needle aspiration, and pleural fluid) with different technical preclinical management protocols and different diagnostic yields. Immunocytochemistry (ICC) has a pivotal role in the determination of diagnostic, prognostic, and predictive markers. Therefore, limited cytology samples are to be used with a cell-sparing approach, to allow both diagnostic ICC evaluation as well as predictive marker assessment by ICC or specific molecular assays. In this review, we describe the most common ICC markers used for the diagnosis and prognostic/predictive characterization of thoracic tumors in different cytological specimens., (© 2019 S. Karger AG, Basel.)
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- 2020
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48. PAX8-GLIS3 gene fusion is a pathognomonic genetic alteration of hyalinizing trabecular tumors of the thyroid.
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Marchiò C, Da Cruz Paula A, Gularte-Merida R, Basili T, Brandes A, da Silva EM, Silveira C, Ferrando L, Metovic J, Maletta F, Annaratone L, Pareja F, Rubin BP, Hoschar AP, De Rosa G, La Rosa S, Bongiovanni M, Purgina B, Piana S, Volante M, Weigelt B, Reis-Filho JS, and Papotti M
- Subjects
- Humans, Oncogene Proteins, Fusion genetics, DNA-Binding Proteins genetics, PAX8 Transcription Factor genetics, Repressor Proteins genetics, Thyroid Neoplasms genetics, Thyroid Neoplasms pathology, Trans-Activators genetics
- Abstract
The hyalinizing trabecular adenoma/tumor is a rare and poorly characterized follicular-derived thyroid neoplasm recently shown to harbor recurrent PAX8-GLIS1 or PAX8-GLIS3 gene fusions. Here we sought to define the repertoire of genetic alterations of hyalinizing trabecular tumors, and whether PAX8-GLIS3 fusions are pathognomonic for hyalinizing trabecular tumors. A discovery series of eight hyalinizing trabecular tumors was subjected to RNA-sequencing (n = 8), whole-exome sequencing (n = 3) or targeted massively parallel sequencing (n = 5). No recurrent somatic mutations or copy number alterations were identified in hyalinizing trabecular tumor, whereas RNA-sequencing revealed the presence of a recurrent genetic rearrangement involving PAX8 (2q14.1) and GLIS3 (9p24.2) genes in all cases. In this in-frame fusion gene, which comprised exons 1-2 of PAX8 and exons 3-11 of GLIS3, GLIS3 is likely placed under the regulation of PAX8. Reverse transcription RT-PCR and/or fluorescence in situ hybridization analyses of a validation series of 26 hyalinizing trabecular tumors revealed that the PAX8-GLIS3 gene fusion was present in all hyalinizing trabecular tumors (100%). No GLIS1 rearrangements were identified. Conversely, no PAX8-GLIS3 gene fusions were detected in a cohort of 237 control thyroid neoplasms, including 15 trabecular thyroid lesions highly resembling hyalinizing trabecular tumor from a morphological standpoint, as well as trabecular/solid follicular adenomas, solid/trabecular variants of papillary carcinoma, and Hurthle cell adenomas or carcinomas. Our data provide evidence to suggest that the PAX8-GLIS3 fusion is pathognomonic for hyalinizing trabecular tumors, and that the presence of the PAX8-GLIS3 fusion in thyroid neoplasms may be used as an ancillary marker for the diagnosis of hyalinizing trabecular tumor, thereby avoiding overtreatment in case of misdiagnoses with apparently similar malignant tumors.
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- 2019
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49. Spread through air spaces (STAS) is a predictor of poor outcome in atypical carcinoids of the lung.
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Altinay S, Metovic J, Massa F, Gatti G, Cassoni P, Scagliotti GV, Volante M, and Papotti M
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- Adenocarcinoma pathology, Adult, Carcinoid Tumor metabolism, Carcinoma, Squamous Cell pathology, Case-Control Studies, Disease-Free Survival, Female, Humans, Lung pathology, Lung Neoplasms pathology, Male, Middle Aged, Neoplasm Invasiveness pathology, Neoplasm Recurrence, Local pathology, Neoplasm Staging, Prognosis, Retrospective Studies, Adenocarcinoma of Lung diagnosis, Adenocarcinoma of Lung pathology, Carcinoid Tumor pathology
- Abstract
Spread through air spaces (STAS) have been recently recognized as a prognostic factor for adenocarcinoma and squamous cell carcinoma of the lung. Pulmonary neuroendocrine neoplasms (NENs) include tumors with different morphology and a heterogeneous clinical behavior. Among atypical carcinoids (ACs), new prognostic factors able to refine prognosis are needed. In the present study, a retrospective series of 91 surgically resected ACs was investigated, in parallel with 191 control cases of typical carcinoids (TCs) and of high-grade small- and large-cell neuroendocrine carcinomas, to assess the presence and potential prognostic role of STAS. STAS was defined by the presence of neoplastic nests or single cells in air spaces beyond the tumor edge. Clinicopathological parameters and survival were correlated by univariate and multivariate analyses. STAS was identified in 48% of ACs (44/91) compared to 20.5% of TCs and 71-88% of high-grade large- and small-cell carcinomas in the control group. In the carcinoid group, presence of STAS was significantly correlated with unfavorable parameters, such as high tumor stage, positive nodal status, high Ki-67 index, presence of angioinvasion, and with adverse disease outcome, shorter overall survival, and time to progression. In conclusion, the presence of STAS is an additional relevant adverse prognostic factor in pulmonary AC that currently has the most unpredictable outcome and the most controversial treatment strategy.
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- 2019
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50. Eccrine spiradenoma of the nipple: Case report, differential diagnosis and literature review.
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Metovic J, Gallino C, Zanon E, Bussone R, Russo R, Vissio E, Annaratone L, Conti L, Papotti M, Cassoni P, and Castellano I
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- Acrospiroma metabolism, Acrospiroma pathology, Adenoma metabolism, Aged, 80 and over, Breast Neoplasms metabolism, Breast Neoplasms pathology, Diagnosis, Differential, Female, Humans, Sweat Gland Neoplasms metabolism, Sweat Gland Neoplasms pathology, Acrospiroma diagnosis, Adenoma diagnosis, Adenoma pathology, Breast Neoplasms diagnosis, Nipples pathology, Sweat Gland Neoplasms diagnosis
- Abstract
Eccrine spiradenoma is a rare lesion originating from eccrine sweat glands, with only few cases reported in breast tissue: we here describe for the first time, an eccrine spiradenoma arising in the nipple. An 84 year-old woman with a lesion enlarging her right nipple, showing ulcerations and eczema-like changes of the covering skin, was admitted to our hospital. Surgical excision of the central quadrant with nipple-areola complex was performed, followed by histopathological evaluation which revealed an adenoma with predominantly basaloid epithelial cells. The lesion was composed of tightly packed small and large groups of cells, arranged in diffuse alveolar/pseudorosette formations. The small cells expressed p63 and calponin, while a positive expression of CK7 and CD117 was detected in large cells. After careful and detailed examination, excluding various similar entities, a diagnosis of eccrine spiradenoma has been rendered. Although extremely rare, eccrine spiradenoma should be taken into account in the differential diagnosis of subcutaneous primary breast tumors.
- Published
- 2019
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