Bioarchaeology aids the cultural understanding of six characters in search of their agency (Tarquinia, ninth-seventh century BC, central Italy).

Etruria contained one of the great early urban civilisations in the Italian peninsula during the first millennium BC, much studied from a cultural, humanities-based, perspective, but relatively little with scientific data, and rarely in combination. We have addressed the unusual location of twenty inhumations found in the sacred heart of the Etruscan city of Tarquinia, focusing on six of these as illustrative, contrasting with the typical contemporary cremations found in cemeteries on the edge of the city. The cultural evidence suggests that the six skeletons were also distinctive in their ritualization and memorialisation. Focusing on the six, as a representative sample, the scientific evidence of osteoarchaeology, isotopic compositions, and ancient DNA has established that these appear to show mobility, diversity and violence through an integrated bioarchaeological approach. The combination of multiple lines of evidence makes major strides towards a deeper understanding of the role of these extraordinary individuals in the life of the early city of Etruria.

Stratigraphic context and paleoenvironmental significance of minor taxa (Pisces, Reptilia, Aves, Rodentia) from the late Early Pleistocene paleoanthropological site of Buia (Eritrea).

The Buia Homo site, also known as Wadi Aalad, is an East African paleoanthropological site near the village of Buia that, due to its very rich yield from the late Early Pleistocene, has been intensively investigated since 1994. In this paper, which reports on the finds of the 2010-2011 excavations, we include new fossil evidence on previously identified taxa (i.e., reptiles), as well as the very first description of the small mammal, fish and bird remains discovered. In particular, this study documents the discovery of the first African fossil of the genus Burhinus (Aves, Charadriiformes) and of the first rodent from the site. This latter is identified as a thryonomyid rodent (cane rat), a relatively common taxon in African paleoanthropological faunal assemblages. On the whole, the new occurrences documented within the Buia vertebrate assemblage confirm the occurrence of taxa characterized by strong water dependence. The paleoenvironmental characteristics of the fauna are confirmed as fully compatible with the evidence obtained through sedimentology and facies analysis, documenting the sedimentary evolution of fluvio-deltaic and lacustrine systems.

Palaeontology: early Neolithic tradition of dentistry.

Prehistoric evidence for the drilling of human teeth in vivo has so far been limited to isolated cases from less than six millennia ago. Here we describe eleven drilled molar crowns from nine adults discovered in a Neolithic graveyard in Pakistan that dates from 7,500-9,000 years ago. These findings provide evidence for a long tradition of a type of proto-dentistry in an early farming culture.

Changes in skeletal robusticity in an iron age agropastoral group: the Samnites from the Alfedena necropolis (Abruzzo, Central Italy).

Cross-sectional geometrical (CSG) properties of an Iron Age Samnite group from the Alfedena necropolis (Abruzzo, Italy, 2600-2400 B.P.) are compared with a Ligurian Neolithic sample (6000-5500 B.P.). In the period under examination, Samnites were organized in a tribal confederation led by patrilinear aristocracies, indicating incipient social stratification. In comparison, Neolithic society lacked clear signs of social hierarchy. The subsistence of both groups was mainly based on pastoralism and agriculture, but changes in habitual behavior are expected due to the socio-economic transformations that characterized the Iron Age. The Samnites' warlike ideology suggests that unimanual weapon-use and training would have become frequent for males. The intensification of agriculture and the adoption of transhumant pastoralism, performed by a smaller subset of the population, likely led to a lower average level of logistic mobility. The strongly genderized ideology of the period suggests a strict sexual division of labor, with women primarily performing sedentary tasks. CSG properties based on periosteal contours were calculated for humeri, femora, and tibiae (N = 61). Results corroborated the expectations: Alfedena males show substantial humeral bilateral asymmetry, indicating prevalent use of one arm, likely due to weapon training. In both sexes lower limb results indicate reduced mobility with respect to the Neolithic group. Sexual dimorphism is significant in both humeral asymmetry and lower limb indicators of mobility. Although both groups could be broadly defined as agropastoral based on archeological and historical evidence, CSG analysis confirmed important differences in habitual behavior.

Nuclear fluorescence serum reactivity on monkey oesophagus: a new antibody for the follow-up of coeliac disease?

We have identified previously a nuclear fluorescence reactivity (NFR) pattern on monkey oesophagus sections exposed to coeliac disease (CD) patients' sera positive for anti-endomysium antibodies (EMA). The aim of the present work was to characterize the NFR, study the time-course of NFR-positive results in relation to gluten withdrawal and evaluate the potential role of NFR in the follow-up of CD. Twenty untreated, 87 treated CD patients and 15 healthy controls were recruited and followed for 12 months. Their sera were incubated on monkey oesophagus sections to evaluate the presence of NFR by indirect immunofluorescence analysis. Duodenal mucosa samples from treated CD patients were challenged with gliadin peptides, and thus the occurrence of NFR in culture supernatants was assessed. The NFR immunoglobulins (Igs) reactivity with the nuclear extract of a human intestinal cell line was investigated. Serum NFR was present in all untreated CD patients, persisted up to 151 ± 37 days from gluten withdrawal and reappeared in treated CD patients under dietary transgressions. Serum NFR was also detected in two healthy controls. In culture supernatants of coeliac intestinal mucosa challenged with gliadin peptides, NFR appeared before EMA. The Igs responsible for NFR were identified as belonging to the IgA2 subclass. The NFR resulted differently from EMA and anti-nuclear antibodies, but reacted with two nuclear antigens of 65 and 49 kDa. A new autoantibody, named NFR related to CD, was described. Furthermore, NFR detection might become a valuable tool in monitoring adherence to a gluten-free diet and identifying slight dietary transgressions.

The p85 regulatory subunit of PI3K mediates TSH-cAMP-PKA growth and survival signals.

Phosphatidylinositol 3-kinase (PI3K) is necessary for thyroid stimulating hormone (TSH)-induced cell cycle progression. To determine the molecular mechanism linking PI3K to TSH, we have identified a serine residue in p85alpha(PI3K) phosphorylated by protein kinase A (PKA) in vitro and in vivo. Expression of an alanine mutant (p85A) abolished cyclic AMP/TSH-induced cell cycle progression and was lethal in thyroid cells (FRTL-5). The aspartic version of the p85alpha(PI3K) (p85D) inhibited apoptosis following TSH withdrawal. The p85alpha(PI3K) wild type not the p85A bound PKA regulatory subunit RIIbeta in cells stimulated with cAMP or TSH. The binding of the aspartic version of p85alpha(PI3K) to RIIbeta was independent of cAMP or TSH stimulation. Similarly, binding of PI3K to p21Ras and activation of AKT, a downstream PI3K target, were severely impaired in cells expressing the p85A mutant. Finally, we found that the catalytic activity of PI3K was stimulated by TSH in cells expressing the wild-type p85alpha(PI3K) but not in cells expressing p85A. This latter mutant did not affect the epidermal growth factor-stimulated PI3K activity. We suggest that (1) TSH-cAMP-induced PKA phosphorylates p85alpha(PI3K) at serine 83, (2) phosphorylated p85alpha(PI3K) binds RIIbeta-PKA and targets PKAII to the membrane, and (3) PI3K activity and p21Ras binding to PI3K increase and activate PI3K downstream targets. This pathway is essential for the transmission of TSH-cAMP growth signals.

Chest Abdominal-Focused Assessment Sonography for Trauma during the primary survey in the Emergency Department: the CA-FAST protocol.

PURPOSE: To evaluate the feasibility of a new protocol, Chest Abdominal-Focused Assessment Sonography for Trauma (CA-FAST), during the primary survey and to estimate its diagnostic accuracy when compared with thoracoabdominal computed tomography (CT) scan. METHODS: A prospective accuracy study was performed from November 2012 to November 2013 at the Emergency Department. Only adult trauma patients who underwent a CA-FAST examination prior to a thoracoabdominal CT scan were enrolled. In addition to standard patterns detected by Extended-FAST (E-FAST) such as pneumothorax (PTX), hemothorax (HTX), pericardial and intraabdominal effusion, CA-FAST protocol also included the research of lung contusions (LCs). RESULTS: Six hundred and one patients were enrolled. The mean time for protocol execution was 7 ± 3 min. Chest ultrasonography showed the following results (all p < 0.001): LCs sensitivity 59 %, specificity 98 %, positive predictive value (PPV) 92 %, negative predictive value (NPV) 86 %, accuracy 87 %; PTX sensitivity 84 %, specificity 98 %, PPV 93 %, NPV 95 %, accuracy 95 %; HTX sensitivity 82 %, specificity 97 %, PPV 87 %, NPV 95 %, accuracy 94 %. The standard 4-views FAST examination showed a diagnostic accuracy of 91 % with a sensitivity of 75 %, specificity of 96 %, PPV of 81 % and NPV of 94 %. CONCLUSION: According to our results CA-FAST protocol proved to be a rapid bedside method, with good accuracy and high NPV in detection of ultrasonographic patterns suggestive of serious injury in trauma patients; moreover, the additional research of LCs did not cause a delay in the diagnosis. Ultrasonography should be used as initial investigation during the primary survey, sending to further diagnostic studies (CT scan) only those patients not clearly classified.

A health assessment of high status Christian burials recovered from the Roman-Byzantine archeological site of Elaiussa Sebaste, Turkey.

The purpose of this study is to assess the state of health of 116 individuals whose remains were excavated from Byzantine period burials underneath the floor of an important Christian basilica from the site of Elaiussa Sebaste, Turkey. Elaiussa Sebaste was a Mediterranean coastal community, which began as a Roman town and continued as an early Christian Byzantine community until the end of the 7th century AD. The burials date from the middle of the 6th through the middle of the 7th centuries AD. We attempt to determine how high social status has influenced the type and frequency of skeletal lesions exhibited in this sample. All strata of this population show a number of chronic and acute health problems as indicated by skeletal lesions. Yet, only the frequency of degenerative joint disease (DJD) differs by sex, with males exhibiting a higher rate of DJD than females, p=0.09. There is no difference in the frequency of trauma among adult males and females. Non-specific skeletal lesions (cribra orbitalia, porotic hyperostosis, and periostitis) often associated with dietary and general stressors, but also with specific systemic diseases, are common in both sexes. The sub-adults primarily exhibit periostitis of the long bones and do not show skeletal lesions specific to malaria. It seems that high social ranking did not prevent serious ailments from affecting the health of individuals living in the Elaiussa Sebaste community.

PARP inhibitors enhance replication stress and cause mitotic catastrophe in MYCN-dependent neuroblastoma.

High-risk and MYCN-amplified neuroblastomas are among the most aggressive pediatric tumors. Despite intense multimodality therapies, about 50% of these patients succumb to their disease, making the search for effective therapies an absolute priority. Due to the important functions of poly (ADP-ribose) polymerases, PARP inhibitors have entered the clinical settings for cancer treatment and are being exploited in a variety of preclinical studies and clinical trials. PARP inhibitors based combination schemes have also been tested in neuroblastoma preclinical models with encouraging results. However, the expression of PARP enzymes in human neuroblastoma and the biological consequences of their inhibition remained largely unexplored. Here, we show that high PARP1 and PARP2 expression is significantly associated with high-risk neuroblastoma cases and poor survival, highlighting its previously unrecognized prognostic value for human neuroblastoma. In vitro, PARP1 and 2 are abundant in MYCN amplified and MYCN-overexpressing cells. In this context, PARP inhibitors with high 'PARP trapping' potency, such as olaparib or talazoparib, yield DNA damage and cell death preceded by intense signs of replication stress. Notwithstanding the activation of a CHK1-CDC25A replication stress response, PARP-inhibited MYCN amplified and overexpressing cells fail to sustain a prolonged checkpoint and progress through mitosis in the presence of damaged DNA, eventually undergoing mitotic catastrophe. CHK1-targeted inhibition of the replication stress checkpoint exacerbated this phenotype. These data highlight a novel route for cell death induction by PARP inhibitors and support their introduction, together with CHK1 inhibitors, in therapeutic approaches for neuroblastomas with high MYC(N) activity.

Transforming growth factor-beta1 down-regulation of major histocompatibility complex class I in thyrocytes: coordinate regulation of two separate elements by thyroid-specific as well as ubiquitous transcription factors.

Transforming growth factor (TGF)-beta1-decreased major histocompatibility complex (MHC) class I gene expression in thyrocytes is transcriptional; it involves trans factors and cis elements important for hormone- as well as iodide-regulated thyroid growth and function. Thus, in rat FRTL-5 thyrocytes, TGF-beta1 regulates two elements within -203 bp of the transcription start site of the MHC class I 5'-flanking region: Enhancer A, -180 to -170 bp, and a downstream regulatory element (DRE), -127 to -90 bp, that contains a cAMP response element (CRE)-like sequence. TGF-beta1 reduces the interaction of a NF-kappaB p50/fra-2 heterodimer (MOD-1) with Enhancer A while increasing its interaction with a NF-kappaB p50/p65 heterodimer. Both reduced MOD-1 and increased p50/p65 suppresses class I expression. Decreased MOD-1 and increased p50/p65 have been separately associated with the ability of autoregulatory (high) concentrations of iodide to suppress thyrocyte growth and function, as well as MHC class I expression. TGF-beta1 has two effects on the downstream regulatory element (DRE). It increases DRE binding of a ubiquitously expressed Y-box protein, termed TSEP-1 (TSHR suppressor element binding protein-1) in rat thyroid cells; TSEP-1 has been shown separately to be an important suppressor of the TSH receptor (TSHR) in addition to MHC class I and class II expression. It also decreases the binding of a thyroid-specific trans factor, thyroid transcription factor-1 (TTF-1), to the DRE, reflecting the ability of TGF-beta1 to decrease TTF-1 RNA levels. TGF-beta1-decreased TTF-1 expression accounts in part for TGF-beta1-decreased thyroid growth and function, since decreased TTF-1 has been shown to decrease thyroglobulin, thyroperoxidase, sodium iodide symporter, and TSHR gene expression, coincident with decreased MHC class I. Finally, we show that TGF-beta1 increases c-jun RNA levels and induces the formation of new complexes involving c-jun, fra-2, ATF-1, and c-fos, which react with Enhancer A and the DRE. TGF-beta1 effects on c-jun may be a pivotal fulcrum in the hitherto unrecognized coordinate regulation of Enhancer A and the DRE.

Increased type II transforming growth factor-beta receptor expression in liver cells during cholesterol challenge.

A large body of evidences implicates transforming growth factor-beta (TGF-beta) in the pathogenesis of atherosclerosis. In this context, TGF-beta receptor dysfunction has been suggested to be relevant. We tested the effect of hypercholesterolemia, a well-known risk factor for atherosclerosis, on liver type II TGF-beta receptor (TbetaR-II) expression in atherosclerosis-susceptible C57BL/6 mouse strain fed atherogenic diet. In addition, the relationship between cholesterol and TbetaR-II expression was verified by cholesterol challenge on human hepatoma cell (HepG2) cultures. The susceptible C57BL/6 mice fed atherogenic diet exhibited significant mRNA and immunohistochemical TbetaR-II liver expression at 2, 5, 9 and 15 weeks as compared to animals fed a regular diet. The TbetaR-II profile on HepG2 resulted in a time-dependent increased expression when the cells were incubated with soluble free cholesterol, associated with an increased TGF-beta-dependent biological activity as detected by luciferase assay of reporter gene. These data provide evidence for a cholesterol-dependent TbetaR-II induction that may play a potentially relevant role in the development of hypercholesterolemia and atherogenesis.

Tumor markers and lung cancer: correlation between serum and bronchial secretion levels of CEA, TPA, CanAg CA-50, NSE and ferritin.

The levels of carcinoembryonic antigeny (CEA), tissue polypeptide antigeny (TPA), CanAg 50, neuron specific enolase (NSE) and ferritin were determined in bronchial secretion and serum of patients with neoplastic and non-neoplastic lung diseases. Simultaneous determination of two or three markers in the serum and in bronchoalveolar lavage (BAL) may be clinically useful for the diagnosis of lung cancer and even for the type of tumor. The positivity of CEA determined simultaneously in serum and in BAL of patients with lung cancer is higher than 80% whereas in patients with benign lung disease it is lower than 40%. The simultaneous assay of TPA in serum and in BAL showed 100% positivity in patients with oat-cell carcinoma, the frequencies of positivity were similar in patients with non-oat-cell carcinoma. For NSE and CanAg CA-50 patients with oat-cell carcinoma showed 100% positivity. Simultaneous assay of ferritin in serum and in BAL gave 85% positivity in patients with oat-cell carcinoma and only 23% in patients with non-oat-cell carcinoma. We conclude that the simultaneous determination of CEA and CanAg CA-50 or NSE in serum and in BAL is a useful aid in the diagnosis of lung malignancy.

Epidermal growth factor receptor and tissue tumor markers in human intracranial neoplasms.

The content of the epidermal growth factor receptor (EGFr) and its relation to tissue CEA and NSE were examined in 22 intracranial neoplasms (18 primary intracranial tumors and 4 metastasis from lung cancer). The EGFr was measured by a radioligand assay on crude plasma membrane preparations. The tissue tumor markers (CEA and NSE) were measured on the high speed supernatant of tissue homogenates. In the primary intracranial tumors the EGFr was positive in 68%, CEA in 43% and NSE in 47% of all cases examined. As it concerns the metastasis from lung cancer the positivity was 75% for EGFr while for CEA and NSE was 50% and 32% respectively. These results suggest that the routine measurement of EGFr, associated with the determinations of tissue CEA and NSE could provide useful information on the type of the intracranial neoplasms.

Oncogenes and antioncogenes involved in human thyroid carcinogenesis.

The development of cancer is due to the accumulation of multiple somatic mutations, in some cases following germline mutations, which occur in hereditary malignancies such as retinoblastomas or multiple endocrine neoplasia (MEN 2A and B). Genetic alterations or changes in the expression of growth regulatory genes can lead to the initiation of malignant transformation and to eventual tumor progression. Cells that have undergone these cumulative alterations in either the structure or expression of these regulatory genes generally possess a selective growth and/or metastatic advantage over other normal non-transformed cells. Thus, activation of dominantly transforming oncogenes by point mutations, gene amplification, chromosomal translocation or insertional mutagenesis can lead to uncontrolled cellular growth or to a disruption in normal differentiation or apoptosis. Equally contributory to the process of malignant progression is the inactivation of recessive tumor suppressor genes due to point mutations and/or loss of heterozygosity in one allele, which can ultimately lead to a reduction of homozygosity in both alleles. Thyroid tumors in humans represent a particularly suitable multistage model of epithelial tumorigenesis. In fact, even though most thyroid neoplasms originate from a single cell type, i.e. the thyroid follicular cell, they include a broad spectrum of tumors with different phenotypic characteristics and variable biological and clinical behaviour. Multiple degrees of malignancies have been defined: from the benign colloid adenomas through the slowly progressive differentiated papillary and follicular carcinomas to the invariably fatal anaplastic carcinomas, although these histological changes are not necessarily sequential. In this review an effort has been made to summarize and integrate new data published on genetic lesions and altered expression of genes involved in the tumorigenesis of the follicular type of thyroid cancer. We have focused our interest only on gene alterations inducing gain or loss of function, that have been studied in vivo in human thyroid tumor specimens by the use of different techniques, such as PCR mediated DNA analyses, sequencing, mRNA level evaluation and protein expression by immunohistochemical staining.

Mach one impact test apparatus utilizing an 18.4-mm-bore gas gun with a novel decelerating device.

The design and operation of a helium-gas-driven gun capable of operation to Mach one is described in this report. The gun utilizes a unique tapered-tube decelerator that captures the sabot and prevents gas discharge from the muzzle. This feature makes it possible to operate the gun in a normal laboratory environment without excessive safety precautions.

"Like a virgin": Absence of rheumatoid arthritis and treponematosis, good sanitation and only rare gout in Italy prior to the 15th century.

OBJECTIVES: This study was conducted to test several hypotheses: 1. That rheumatoid arthritis and syphilis were New World diseases, only transmitted to the Old World subsequent to the passages of Christopher Columbus; 2. To indirectly test the hypothesis that lead poisoning was prevalent in Roman Italy by looking for its byproduct, gout; 3. To test the hypothesis of compromised sanitation in ancient Italy, on the basis of spondyloarthropathy frequency; and 4. To assess variation of trauma frequencies in ancient Italy, by examining frequency of focal periosteal reaction. METHODS: Skeletons from sites ranging from the Bronze Age to the Black Plague epidemic of 1485-1486 were macroscopically evaluated for focal periosteal reaction and for the cardinal signs of rheumatoid arthritis, treponemal disease, gout and spondyloarthropathy. RESULTS: Examination of 688 individuals revealed low frequency of focally distributed periosteal reaction (bumps) in sites dated from the 3400-700 years before present, sharply increasing in the 15th century. Diffuse periosteal reaction was present only as isolated occurrences secondary to hypertrophic osteoarthropathy and sabre shin reaction was notably absent. Erosive disease was uncommon and always oligoarticular in distribution. No marginal erosions were present, with the exception of an isolated metatarsal with classic overhanging edge sign of gout. Subchondral erosions, peripheral joint fusion and axial skeletal involvement identified spondyloarthropathy frequencies of 1-3%, independent of the antiquity of the site. CONCLUSIONS: Italy, prior to Columbus was like a virgin. Rheumatoid arthritis and treponemal disease (specifically syphilis) were not present, further supporting the contention that they are New World-derived diseases. Periosteal signs of minor trauma were rare prior to fifteenth century plague times. This suggests a potential role of domestic (as opposed to outside environment activities) in is development. The hypothesis for a role of lead poisoning in the demise of the Roman Empire is falsified by the rarity of gout. The frequency of spondyloarthropathy was significantly below that found in sanitarily challenged populations, suggesting high standards of hygiene in ancient Italy.

Seasonality of marriages and ecological contexts in rural communities of central-southern Italy (Abruzzo), 1500-1871.

The seasonality of 27,705 marriages celebrated in a four century span in the Province of Teramo (Abruzzo, Italy) was analyzed to identify the presence of a long-term pattern related to the prevailing subsistence activity and the main factors affecting it. The results show general agreement in all centuries with the agricultural patterns of other lowland or south-central Italian groups (Piedmont, Veneto, Liguria, Emilia Romagna, Tuscany, Latium, Compania, Apulia) and Mediterranean regions (Spain, France), although with some differences. Religious factors strongly affected the timing of marriage only during Lent. Of interest is the progressive increase through the centuries of marriages in the summer-autumn months, associated with a decrease in January and February. This suggests the passage from a summer migration system to a rural sedentary system with occasional seasonal work.

PDR-1/hParkin negatively regulates the phagocytosis of apoptotic cell corpses in Caenorhabditis elegans.

Apoptotic cell death is an integral part of cell turnover in many tissues, and proper corpse clearance is vital to maintaining tissue homeostasis in all multicellular organisms. Even in tissues with high cellular turnover, apoptotic cells are rarely seen because of efficient clearance mechanisms in healthy individuals. In Caenorhabditis elegans, two parallel and partly redundant conserved pathways act in cell corpse engulfment. The pathway for cytoskeletal rearrangement requires the small GTPase CED-10 Rac1 acting for an efficient surround of the dead cell. The CED-10 Rac pathway is also required for the proper migration of the distal tip cells (DTCs) during the development of the C. elegans gonad. Parkin, the mammalian homolog of the C. elegans PDR-1, interacts with Rac1 in aged human brain and it is also implicated with actin dynamics and cytoskeletal rearrangements in Parkinsons's disease, suggesting that it might act on engulfment. Our genetic and biochemical studies indicate that PDR-1 inhibits apoptotic cell engulfment and DTC migration by ubiquitylating CED-10 for degradation.

Cranial trauma in iron age Samnite agriculturists, Alfedena, Italy: implications for biocultural and economic stress.

The Samnites are an Iron Age protohistoric people from the central region of Italy. The skeletal remains are from the Alfedena necropolis, 6th through 5th centuries B.C. Macchiarelli et al. (Antropologia Contemporanea 4 (1981) 239-243) were the first to report on cranial trauma for this population, presenting four cases with extreme injuries. We re-examined this well documented skeletal population for additional examples of trauma. Previously unexamined remains from Alfedena, excavated at the turn of the 20th century, are also included in our analysis (Mariani. 1901. "Aufidena", ricerche archeologiche e storiche del Sannio settentrionale. Roma: Acc Naz Dei Lincei). Of the 209 adult crania examined, 12.9% of them exhibited trauma. Analysis of location and frequency of cranial trauma revealed that cranial injuries to the head appear to originate from all directions. The high rate of cranial trauma underscores the violent circumstances experienced during the Iron Age protohistoric period of central Italy. Males are much more likely to exhibit cranial injury than females (P = 0.009). We conclude that the injuries received by Samnite male farmer-warriors occurred while defending pastoral-agricultural resources. Trauma rates are similar for some Iron Age populations and not for others. Behavior associated with violence during the Iron Age period can not be generalized for all populations found in Italy.