[Antineoplastic profile assessment for the evaluation of occupational exposure in hospital personnel involved in handling cytotoxic agents]. / Definizione del profile di chemioterapici antiblastici nella valutazione dell'esposizione professionale in ambito sanitario.

Environmental and biological monitoring performed in health care settings during a 14-year period allowed numerous data to be obtained by using the HPLC coupled with tandem mass spectrometry (HPLC-MS/MS). Data was stored in a specific data-base for the assessment of risk exposure to antineoplastic agents (CA). The strategy of analysis was based on the simultaneous determination of different active substances with a lower limit of quantification (LOQ) optimized in the range of sub-units of microg/L. In the present study, the MRM-ESI-MS/MS profiles of seven antineoplastic agents in both environmental and biological matrices are reported. These methods validated according to FDA guidelines allowed our lab to define a profile of antineoplastic agents that was representative of the four IARC classes, such as cyclophosphamide (group 1), cisplatin and doxorubicin (group 2A), daunorubicin (group 2B), 5-fluorouracil, ifosfamide (group 3), taxol, and gemcitabine (group 4). Moreover, contamination levels on a number of work surfaces and trends over a 14-year period are presented. The evaluation of occupational exposure to CA has been based on ALARA principle for most of the past decades but this principle is nowadays overwhelmed by the fast development of technology. The assessment of a possible in-take of CA in hospital personnel, when the sources of environmental contamination are identified, has become possible by the defining of a limit of exposure close to the limit of detection of the analytical method.

[Headache as "sentinel" symptom in personnel involved in the preparation and administration of antineoplastic drugs]. / La Cefalea sintomo "sentinella" nel personale addetto alla preparazione e somministrazione di Chemioterapici Antiblastici.

The increased prevalence of neoplastic diseases observed over the last years has resulted for in more frequent operations of preparation and administration of antiblastic drugs performed by the medical personnel. In this study, we examined a group of subjects involved in the preparation of ACNU, Methotrexate, Novantrone, Vincristine, Cyclophosphamide, Carboplatinum, Mythoxantrone, in order to elucidate whether headache may represent an early symptom of exposure to these products. At the same time, we measured the degree of pollution in the air surrounding the vertical laminar flow aspiration cabinet used for drug preparation. The 66.6% of the 12 subjects studied, 9 females and 3 males, complained of headache. However, the environmental detection using high performance liquid chromatography coupled with triple quadrupole mass spectrometry (HPLC/MS/MS) gave negative results with respect to a possible environmental damage. These findings suggest that headache is the onset symptom of the toxic effect of antiblastic chemiotherapics in the medical personnel involved in drug preparation and administration.

Determination of 5-fluorouracil in environmental samples by solid-phase extraction and high-performance liquid chromatography with ultraviolet detection.

5-Fluorouracil (5-FU) is one of the most widely used antineoplastic drugs. It can be therefore considered to be a model compound for the identification of exposure routes during preparation and administration of cytostatic agents, especially for nucleoside analogue drugs. In this study, an HPLC-UV method was validated for determination of 5-FU in wipe samples by direct analysis of the aqueous solutions and in air samples by using solid-phase extraction (SPE). When samples were pre-treated on styrene-divinylbenzene resin SPE columns, a 20-fold preconcentration of the analyte was achieved. As regards air samples, correlation coefficients were always higher than 0.998 and the limit of detection was assessed at 15 ng on filter. In order to verify the reliability of these procedures, 5-chlorouracil was used as internal standard. The procedure presented here has been applied to the environmental monitoring of occupational exposed subjects. The amount of 5-FU ranged from 0.043 to 0.23 microg/m3 in air samples and from 0.2 to 470.1 microg/dm2 in wipe samples. 5-FU was also detected on the internal side of the gloves (0.07 to 3.77 microg/pair of gloves).

Rapid and sensitive determination of paclitaxel (Taxol(R)) in environmental samples by high-performance liquid chromatography tandem mass spectrometry

A new, rapid and sensitive method for the quantitative determination of paclitaxel (Taxol(R)) in environmental samples is developed and validated. This highly sensitive and specific method was successfully applied to an environmental monitoring study for identifying occupational exposure situations to anticancer drugs. After addition of an internal standard, 2'-methylpaclitaxel, aqueous extraction of wipe samples or air filters followed by a single-step liquid-liquid extraction with ethyl acetate was performed. Extraction efficiency averaged 90%. The method is based on analysis by liquid chromatography with tandem mass spectrometry in selected reaction monitoring mode of environmental samples from the preparation and administration of cytostatic agents. The assay is linear (r = 0.998) in the range 2-64 ng on filters (air samples), and 20-320 ng on wipe samples. The accuracy of the assay is always

Determination of methotrexate in environmental samples by solid phase extraction and high performance liquid chromatography: ultraviolet or tandem mass spectrometry detection?

In order to assess exposure levels of hospital personnel involved in the preparation and administration of antineoplastic drugs, environmental monitoring should be carried out. Wipe samples, pads, gloves and air samples should be collected at the end of each work shift, properly treated and then analysed using instrumental techniques which are sufficiently sensitive and specific to detect even trace amounts of drug. In this study, a method using high performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS), incorporating solid phase extraction (SPE), was validated for determination of methotrexate (MTX) in wipe and air samples. Each step of the method was first developed and optimised using ultraviolet detection (UV), and afterwards tandem mass spectrometry was used to obtain a lower limit of quantitation when the expected drug level was less than the analytical UV detection limit. SPE enabled a 20-fold preconcentration of the analyte when using HPLC/UV and a further 30-fold preconcentration was obtained when analysing samples by HPLC/MS/MS. For example, the limit of quantitation (LLQ) was lowered from 3000 ng on wipe (direct injection onto an HPLC/UV system) to 5 ng on wipe (SPE plus HPLC/MS/MS). 7-hydroxymethotrexate was used as internal standard to assess precision and accuracy.

Seasonal effect on airborne pyrene, urinary 1-hydroxypyrene, and benzo(a)pyrene diol epoxide-hemoglobin adducts in the general population.

Exposure to airborne polycyclic aromatic hydrocarbons (PAHs) in 65 employees (40 sampled both in summer and winter, 15 sampled in summer only, and 10 sampled in winter only) with no occupational exposure to PAHs was assessed by measuring: personal exposure to pyrene, urinary excretion of 1-hydroxypyrene (1-OHP), and benzo(a)pyrene diol epoxide adducts to hemoglobin (BPDE-Hb). Overall, office employees were exposed to significantly higher levels of pyrene in winter (4.54 +/- 2.35 ng/m3, mean +/- SD) than in summer (1.67 +/- 1.92 ng/m3, mean +/- SD; P < 0.001), but no such seasonal variability was observed in 1-OHP excretion. Tobacco smoking was the major determinant of 1-OHP excretion. BPDE-Hb adducts were measured by gas chromatography-mass spectrometry as benzo(a)pyrene tetrols (BPT) released from adducted hemoglobin. In the 65 employees analyzed, mean BPT levels +/- SD were higher in winter (0.14 +/- 0.38 fmol/mg Hb) than summer (0.031 +/- 0.022 fmol/mg Hb). This difference was not statistically significant, probably because of the small proportion of subjects with detectable adducts (11% in summer and 16% in winter). BPDE-Hb adducts were not significantly associated with sex, age, diet, smoking habits, or with pyrene levels and 1-OHP excretion. This is the first report providing reference BPDE-Hb adduct values for the general population not occupationally exposed to environmental PAHs and shows a tendency to seasonal variability, with higher BPT levels in winter when environmental PAHs are also high.

Urinary 1-hydroxypyrene as a marker of exposure to pyrene: an epidemiological survey on a general population group.

Urinary levels of 1-hydroxypyrene in a general adult population group are studied. Experimental data are not normally distributed; statistical analysis required a base 10 logarithmic transformation of data. The concentrations of urinary 1-hydroxypyrene measured were expressed as microgram g-1 urinary creatinine and are comparable with those reported by other authors, both for smoker and non-smoker subgroups. Multiple regression analysis shows that, for smokers, the number of cigarettes smoked per day and the body mass index (BMI) significantly influence the levels of urinary 1-hydroxypyrene expressed as microgram g-1 urinary creatinine, whereas no personal or behavioural variable (age, sex, alcohol consumption, dietary intake of pyrene, BMI) modified the 1-hydroxypyrene levels for non-smokers.

Determination of environmental reference concentration of six PAHs in urban areas (Pavia, Italy).

We propose a sampling strategy, using individual dosimetry to measure the daily inhaled quantity of PAHs in urban air. The method was applied to monitor 56 subjects living in an Italian town (Pavia; 80 000 inhabitants) and the Environmental Reference Concentration (E.R.C.) of six PAHs (classified as 'possible' carcinogenic agents for humans) was determined. The individual environmental samplings took place in two different seasons (winter and summer), for persons living in four different urban areas with different traffic density. Subjects were selected using a specific questionnaire designed to collect information on indoor and indoor+outdoor exposure times. The mean +/- S.D. value of Benzo[a]pyrene [BaP] was 0.37 +/- 0.15 ng m-3 in winter and 0.12 +/- 0.07 ng m-3 in summer. Assuming 18 m3 as the daily inhaled quantity the estimate of the BaP inhaled quantity was 6.66 ng/day in winter and 2.16 ng/day in summer.

[Granular cell tumor of the male breast. Case report and review of the literature]. / Tumore a cellule granulari della mammella maschile. Descrizione di un caso e revisione della letteratura.

The authors report a case of granular cell tumor of the breast arising in a 38-year old man. Preoperative diagnosis was of breast carcinoma. However, frozen sections showed the benign nature of the tumour. It is important to identify this lesion as, in spite of a clinical and mammographic similarity with carcinoma, its behaviour is quite benign, and a simple tumour excision is the recommended treatment.

[Primary embryonal rhabdomyosarcoma of the breast. Description of a case]. / Rabdomiosarcoma embrionale primitivo della mammella. Descrizione di un caso.

Authors report a case of embryonal rhabdomyosarcoma of the breast occurring in a 16-year-old girl. To date, 26 cases have been described in the literature of which only 4 in girls younger than 16 years. Histological features, immunohistochemical findings and differential diagnosis are discussed.

[Late pancreatic metastasis of renal carcinoma. Description of 2 cases and review of the literature]. / Metastasi pancreatica tardiva di carcinoma renale. Descrizione di due casi e revisione della letteratura.

Authors report two cases of pancreatic metastasis from renal clear cell carcinoma, discovered in a 62 years-old-man and in a 73 years-old woman, respectively 17 and 11 years after undergoing curative nephrectomy for primary tumor. Immunological forces and slow doubling times of tumor may explain this type of late recurrence, whereas, the endocrine status is lacking in importance. The risk of a recurrence of tumour ten years or later after nephrectomy suggests a lifelong follow-up of patients with an history of renal cell carcinoma.

[Infiltrating lobular carcinoma of the breast, alveolar variant, with stromal reactive osteoclast-like giant cells. Description of a case]. / Carcinoma lobulare infiltrante della mammella, variante alveolare, con cellule giganti reatitive stromali osteoclasto-simili. Descrizione di un caso.

Authors report a case of alveolar variant of infiltrating lobular carcinoma of the breast with stromal osteoclast-like giant cells. Immunohistochemically the carcinoma cells were reactive for EMA and Cytokeratins; osteoclast-like giant cells (OCL-GC) were negative. OCL-GC and histiocytoid mononuclear cells with nuclei similar to those within the OCL-GC present in this case, were strongly immunoreactive with KP-1, a recently developed monoclonal antibody directed against cells of mononuclear phagocytic lineage in routinely processed tissue section and showed immunoreactivity to vimentin. These findings of our case provide evidence against a carcinoma cells derivation and support a monocytic/histiocytic origin for the OCL-GC.

Ductal adenoma of the breast. An immunohistochemical study of five cases.

The immunohistochemical features of 5 ductal adenomas of the breast were investigated. The tumours were composed by tubular glands lined by cells with both luminal and basal/myoepithelial immunophenotypes. Tubules were encircled by an intact basement membrane as demonstrated by staining for laminin and type IV collagen. Areas of dense fibrosis, a usual finding in these lesions, stained for fibronectin, type III collagen and contained vimentin and actin positive spindle cells, consistent with myofibroblasts. These observations confirm a double epithelial and myoepithelial differentiation in these lesions and seem to support the suggestion that ductal adenomas result from sclerosis of ductal papillomas.

Carcinosarcoma of the oesophagus with rhabdomyoblastic differentiation.

We report a case of oesophageal carcinosarcoma with prominent rhabdomyoblastic differentiation. Immunohistochemically, the rhabdomyblasts were mainly reactive to vimentin, cytokeratin, desmin, muscle-specific actin, myosin, and myoglobin, and were surrounded by laminin and type IV collagen-positive basement membranes. The tumour had dual differentiation, carcinomatous and sarcomatous, but also showed evident features of transition between the two components; suggesting a common origin. An epithelial-mesenchymal conversion could be the pathogenetic mechanism involved in the histogenesis of this lesion. The word carcinosarcoma, from a descriptive point of view, seems the most suitable to describe a tumour composed of both carcinomatous and true sarcomatous elements.

The basement membranes in sarcomatoid carcinomas. An immunohistochemical study.

AIMS: Eight sarcomatoid carcinomas from various anatomical locations were investigated by immunohistochemical staining to laminin, type IV collagen and heparan sulfate proteoglycan, to study the characteristics of basement membranes at the interface between carcinomatous and sarcomatous tissues. METHODS: Paraffin wax embedded tissue sections from representative tumor samples have been stained with specific antibodies, using the peroxidase-antiperoxidase technique. RESULTS: In all cases several interruptions or discontinuities of the basement membrane staining pattern were seen. In 4 cases, larger defects or complete loss of staining was also noted. At these places, the boundaries between carcinomatous and sarcomatous tissue were often blurred. CONCLUSIONS: Disruption and loss of basement membranes at interface between carcinomatous and sarcomatous tissues is a frequent finding in sarcomatoid carcinomas. These changes could be consistent with an epithelial origin of the sarcomatous component in these tumors by means of an epithelial-mesenchymal conversion mechanism.

The extracellular matrix in sarcomatoid carcinomas of the breast.

The distribution of the main extracellular matrix components has been investigated immunohistochemically in four cases of breast sarcomatoid carcinomas. The histogenesis of these tumours is still unclear, but most evidence suggests that the sarcomatous component originates from mesenchymal conversion of carcinomatous cells. We found that carcinomatous portions of the tumours were associated with linear basement membrane-like deposition of laminin, type IV collagen and heparan sulphate proteoglycan that partially circumscribed the epithelial nests. The sarcomatous components produced an extracellular matrix immunoreactive to fibronectin, type I, III and VI collagens and tenascin. However, in two cases, in some sarcomatous areas, focal pericellular staining for antibodies to laminin and type IV collagen was seen. These results indicate that modifications observed in the double tissue component of these tumours involve not only the cell shape and the cytoskeleton, but also the components of the extracellular matrix. The significance of these findings and their relevance in the understanding of the phenotypic pattern changes of these biphasic tumours are discussed.

Xanthogranulomatous gastritis: association with xanthogranulomatous cholecystitis.

A case of xanthogranulomatous inflammation of the gastric wall is reported. The lesion was associated with adherent xanthogranulomatous cholecystitis and simulated, clinically, a malignant neoplasm. Histologically, foamy histiocytes, multi-nucleated giant cells, other inflammatory cells, fibrous reaction with spindle cells and cholesterol clefts were found.

[A case of desmoplastic malignant melanoma]. / Su di un caso di melanoma maligno desmoplastico.

Desmoplastic malignant melanoma: report of a case. Authors report a case of desmoplastic malignant melanoma, a variant of the common primary cutaneous malignant melanoma in a 42 years old man, who was admitted into Hospital for metastatic node disease. Differential diagnoses on the primary, eventually discovered on the right knee, are discussed.

Smooth muscle contraction bands in intestinal infarction.

We studied microscopic sections of 24 cases of intestinal infarction looking for contraction bands (CB) in the muscularis propria of the bowel wall. Controls were 11 surgical and 11 autopsy cases from patients who did not suffer any form of ischaemic disease. The difference of CB frequency was significant (P less than 0.001) in infarction versus the surgical control group. Moreover, within the study group the CB frequency was also related to the severity of the ischaemic lesion (P less than 0.01). With immunostaining, CB were not reactive with antibodies against vimentin, desmin, actin or myosin. We propose that CB genesis in intestinal smooth muscle is related to hypoxia, possibly through altered homeostasis of calcium and catecholamine metabolism.