Rosalba Ciranni

Benzene and the genotoxicity of its metabolites. II. The effect of the route of administration on the micronuclei and bone marrow depression in mouse bone marrow cells

Benzene (880 mg/kg) and 4 of its metabolites, i.e., phenol (265 mg/kg), hydroquinone (80 mg/kg), catechol (40 mg/kg), and p-benzoquinone (5-20 mg/kg) have been tested for their capability to induce micronuclei in bone marrow cells of male mice after oral administration or intraperitoneal injection. Oral administration of benzene shows more activity than intraperitoneal injection, whereas the metabolites show more activity if administered by the latter method. The respective genotoxic strengths of the benzene metabolites are the following: hydroquinone much greater than phenol greater than catechol = p-benzoquinone. This last is active when administered orally.

Vanadium salts induce cytogenetic effects in in vivo treated mice.

Three vanadium salts, vanadyl sulfate (SVO5), sodium orthovanadate (Na3VO4) and ammonium metavanadate (NH4VO3), were tested for induction of genotoxic effects in bone marrow of mice following intragastric treatment. Micronucleus (MN) induction in polychromatic erythrocytes (PCEs), structural (sCA) and numerical (nCA) chromosome aberrations in bone marrow cells were evaluated. The micronucleus test, performed at different harvesting times (0-72 h), was found to be positive for all compounds tested. In contrast, except for vanadyl sulfate, no difference was found between controls and treated animals in the sCA test performed 24 and 36 h after treatment. At the same sampling intervals, second metaphases (M II) were positively scored for nCA induction for all three vanadium salts. In addition, the frequency of hypoploid and hyperploid cells was shown to be statistically different from the control value. Polyploid cells were also induced by all compounds, but their frequency was not statistically significant. The positive results obtained by nCA analysis support the finding of a significant presence of types of micronuclei that are probably aneuploidy-related. This finding was further supported by the successful classification of such micronuclei on the basis of shape and size according to Tinwell and Ashby (1991) during microscope analysis.

Clastogenic effects of hydroquinone: induction of chromosomal aberrations in mouse germ cells

The clastogenic activity of hydroquinone (HQ) in germ cells of male mice was evaluated by analysis of chromosomal aberrations in primary spermatocytes and differentiating spermatogonia. In the first experiment with treated spermatocytes the most sensitive stage of meiotic prophase to aberration induction by HQ was determined. Testicular material was sampled for microscopic analysis of cells in diakinesis-metaphase I at 1, 5, 9, 11, and 12 days after treatment with 80 mg/kg of HQ, corresponding to treated diplotene, pachytene, zygotene, leptotene and preleptotene. The frequencies of cells with structural chromosome aberrations peaked at 12 days after treatment (p less than 0.01). This indicates that the preleptotene when DNA synthesis occurred was the most sensitive stage of meiotic prophase. In the second experiment the dose response was determined 12 days post treatment by applying 2 additional doses of 40 mg/kg and 120 mg/kg. The clastogenic effects induced by 40 and 80 mg/kg were significantly different from the controls (p less than or equal to 0.01) and higher than the results obtained with 120 mg/kg of HQ. A humped dose-effect relationship was observed. In a third experiment the same doses were used to analyse chromosomal aberrations in dividing spermatogonia of mice 24 h after treatment with HQ. All the administered doses gave results statistically different from the control values (p less than or equal to 0.01) and the data were fitted to a linear equation. HQ was found to be clastogenic in male mouse germ cells. It is concluded that the clastogenic effect in male germ cells is of the same order of magnitude as in mouse bone marrow cells.

K-ras mutation in the tumour of King Ferrante I of Aragon (1431–1494) and environmental mutagens at the Aragonese court of Naples

The autopsy of the mummy of Ferrante I of Aragon, King of Naples (1431–1494) revealed an adenocarcinoma extensively infiltrating the muscles of the small pelvis. A molecular study of neoplastic tissue observed a typical mutation of the K-ras gene codon 12: the normal sequence GGT (glycine) was altered into GAT (aspartic acid). The recorded genetic change represents the most frequent mutation of the K-ras gene in sporadic colorectal cancer and is characteristic of the effects of alkylating agents. Recent studies have focused the importance of some alkylating agents, such as the endogenous N-nitroso compounds (NOC). Increased intake of red meat, such as beef, lamb or pork, induces a significant threefold increase of faecal NOC levels, with a range of exposure in faeces similar to that from tobacco-specific NOC in cigarette smoke. The study of alimentary regimens of the Italian Renaissance courts, and in particular of the Aragonese court of Naples, produced evidence of very high red meat consumption, attested also by the palaeonutritional data. Therefore, the alimentary ‘environment’ of the Neapolitan court of the 15th century, with the abundance of natural endogenous alkylating agents, can well explain the K-ras mutation causing the tumour that killed the Aragonese king over five centuries ago. Copyright

Juvenile cirrhosis in a 16th century Italian mummy. Current technologies in pathology and ancient human tissues.

The natural headless mummy of a young man from the Basilica of Saint Domenico Maggiore in Naples (16th century) showed at autopsy a well-preserved fibrous liver with a nodular surface, suggesting a case of cirrhosis. Stereo and light microscope study confirmed this diagnosis. To identify the possible etiology of this cirrhosis, additional techniques currently used in pathology were performed. Hemochromatosis and alpha1-antitrypsin deficiency were investigated without results. Investigation regarding Wilson’s disease gave positive results, since the use of rhodamine staining, which is specific to detect the presence of copper in tissues, resulted in red-brown grains at light microscopy. The positive rhodamine test was invalidated by atomic absorption spectroscopy (AAS), which revealed normal copper levels in the tissues. These negative results and the clear and diffuse macronodularity of the liver suggest a case of post-necrotic cirrhosis.

Goiter in an eighteenth-century Sicilian mummy.

Goiter is still a frequent pathological condition of the thyroid gland. Goiter consists of an enlargement of the gland caused by several physiopathologic events, the most important of which is deficient intake of iodine. A series of eighteenth-century mummies housed in the church of Santa Maria della Grazia in Comiso includes one individual showing a pathological condition, with a very large swelling in the antero-inferior region of the neck. X-rays showed small scattered radiopaque foci. Routine histology showed a large number of circular follicles immersed in fibrous tissue. Finally, immunohistochemistry revealed a strong reactivity for thyroglobulin. The data confirmed the nature of the tissue as thyroid, and the macroscopic diagnosis is that of thyroid goiter.

A forgotten Italian pathologist: Angelo Maffucci (1845–1903) and his scientific thought

Sir, it often happens that for unknown reasons personages of great talent, culture, and humanity remain disowned by the world although they have contributed with their work to the evolution of knowledge. Angelo Maffucci was one of these people (Fig. 1). He was born in 1845 in Calitri, (Avellino, Italy), from a well-off farming family. In 1873, he graduated in medicine at the University of Naples and was awarded the Medal of Merit for his active participation in the struggle against cholera at the Hospital for the Incurable. Two years later, in 1875, he started his scientific work as surgeon and pathologist at the Institute of Pathology of the same University [1]. In that period, he described in detail a disease affecting both the cartilage and blood vessels of a woman who had been hospitalized there. She had never suffered from any important diseases, but her body was deformed as a result of skeletal malformations due to the presence of more than 80 tumors that were large: “from a pea [...] to a small orange, to a foetus head” and affected her arms, hands, left humerus, legs, and feet. The patient suffered from frequent and severe haemorrhages caused by the malformed vessels and underwent amputation of one of her arms. Unfortunately, the stump gangrened and soon after the woman died of pyemia [2]. Maffucci performed autopsy and histological examination of the body, and described all the features of this disorder characterized by benign overgrowths of the cartilage (enchondromatosis), and by dark red irregularly shaped patches of skin (multiple cavernous hemangiomas). He reported the case in a manuscript illustrated with a series of handmade sketches and published in an Italian medical journal in 1881 (Figs. 2 and 3) [2]. The disease in question is quite rare (less than 200 cases are reported in literature) and arises during childhood (4–5 years), independent of race and sex. The etiology is unclear, but genetic involvement of the mesenchymal cells has been hypothesized recently [3]. Maffucci’s descriptions are still topical: the venous malformation appeared in the form of soft bluish nodules made up of a spongy matrix organized in large cavernous vascular spaces, full of blood and separated by connectival stroma; the enchondromas seemed to develop from residues of hyaline cartilage, which underwent a process of proliferation and growth [2, 3]. Sixty years later, Alice Carleton called the disease “Maffucci’s Syndrome” [4]. Angelo Maffucci was, for a short period, Professor of Pathology at the University of Catania and, in 1884, when the Faculty of Medicine of the University of Pisa inaugurated the new chair of pathology, Maffucci was appointed first director. In Pisa, he founded the Museum and the Institute of Human Pathology where he started his successful career as experimental researcher and teacher. Under his guide, Pisa was probably the first Italian University where students had the possibility to associate theoretical instruction with practical activity [1]. One of his most important experimental scientific contributions was in the field of infectious embryopathology of the anthrax and of tuberculosis [5–7]. In the course of his experiments carried out on fowls, he isolated Virchows Arch (2006) 449:495–497 DOI 10.1007/s00428-006-0269-3

The aortic coarctation and the Etruscan man: morphohistologic diagnosis of an ancient cardiovascular disease

Reports of cardiovascular diseases in ancient time are very rare since the material mainly consists of skeletal remains; therefore, these diagnoses can only be carried out indirectly, through the marks left on bones. Here we show a case of coarctation of the aorta diagnosed on bones. Aortic coarctation is a congenital disorder in which a portion of the aorta is narrowed to various extent. A collateral circulation system is enrolled to allow adequate compensation of the blood flow. Collateral vessels may become enlarged, producing a distinctive notching on the pleural surface of the ribs and on adjacent bones. Excavation of a sixth to fifth century B.C. Etruscan tomb revealed three funerary chambers (celle) housing 14 skeletal remains of adults. The ribs of one of the male skeleton showed “nail stroke” indentations. Detailed macroscopic examination enabled us to identify them as notching and led to the diagnosis of postductal aortic coarctation. Histological analysis of bone tissue from the notching areas excluded inflammatory and pathological erosive events, supporting the macroscopic diagnosis. The present paper is the first description of aortic coarctation in paleopathology.

A New Case of Ancient Restoration on an Egyptian Mummy

A new example of restoration performed by the ancient Egyptian embalmers on a mummified body is reported. An imaging study of the mummy labelled 2343 in the Archaeological Museum of Naples has revealed the substitution of an artificial wooden ‘prosthesis’ for missing feet. The mummy probably dates to the Ptolemaic-early Roman Period, when other examples of prosthetics occur.

Prostatic hyperplasia in the mummy of an Italian renaissance prince

Pandolfo III Malatesta (1370–1427) was a leading figure of the Italian Renaissance. He was a valiant soldier and horseman with a very active life style. Historical sources report that he died of a fever in 1427. The tomb, containing his mummified body, has recently been discovered in Fano (Marche, Central Italy).