Expression of Fas and Fas ligand in human testicular germ cell tumours.

In the present study, we analysed the expression of Fas ligand (FasL) and its cognate receptor Fas in 14 seminomatous testicular germ cell tumours (TGCT) and six normal testicular tissues obtained following orchiectomy. Tissue samples have been processed to prepare either total RNA or protein extracts or fixed and embedded in paraffin for immunohistochemistry (IHC) experiments. Quantitative RT-PCR experiments demonstrated in TGCT a significant (p < 0.01) increase of the FasL mRNA expression of 21.1 +/- 5.4 fold, with respect to normal tissues. On the contrary, in the same cancer tissues, the levels of Fas mRNA were significantly (p < 0.01) reduced to 0.27 +/- 0.06 fold. These observations were confirmed in western blot experiments showing a significant increase of FasL and a concomitant decrease of Fas proteins in testicular cancer tissues, with respect to normal testis. Moreover, IHC experiments showed a strong FasL immuno-reactivity in six out of eight TGCT samples analysed, while Fas immuno-positivity was found in cancer cells of only two TGCT tissues. In addition, in all tumour samples, infiltrating lymphocytes were Fas positive. However, no correlation could be observed between Fas or FasL mRNA variations and clinical parameters such as patient's age, TNM stage or tumour size. We also compared the serum levels of soluble FasL (sFasL) of 15 patients affected by seminomatous TGCT, of four patients with non-seminomatous TGCT and six age-matched healthy males. No significant differences in sFasL serum level could be identified. In conclusion, our data demonstrated that the majority of seminomas are characterized by an increased expression of FasL and a concomitant reduction of Fas, with respect to human normal testis, and that sFasL serum level is not a tumour marker for patients affected by TGCT.

Pheochromocytoma associated with adrenocortical tumor in the same gland. Two case reports and literature review.

Pheochromocytomas are catecholamine-producing neuroendocrine tumours arising from chromaffin cells of the adrenal medulla or extra-adrenal paraganglionic system that show 2 distinctive features, rarity and clinical variability. Pheochromocytoma occasionally is associated with pathological lesions of the adrenal cortex. We present 2 cases of patients referred to our hospital with a finding of clinical suspected pheochromocytoma. Both of them were hypertensive; the first patient with typical symptoms of pheochromocytoma and the second patient with chest pain and hypertension resistant to pharmacological treatment. The diagnosis of pheochromocytoma was confirmed in both cases with laboratory analysis and the lesion was achieved by employing 3 imaging techniques: computed tomography (CT), magnetic resonance imaging (MRI) and scintigraphy with (123)I-metaiodobenzilguanidine (MIBG). The patients underwent adrenalectomy and in the same adrenal gland we found a pheochromocytoma associated with a nonfunctioning cortical adenoma. As far as we know few cases with this association are available in the literature.

Carotid endarterectomy and gliofibrillar S100b protein release.

Increased levels of the gliofibrillar S100b protein can be detected during carotid endarterectomy (CEA). Whether the S100b protein increase is marker of brain ischemic sufferance and predictor of cognitive decline is controversial. Twenty-eight patients underwent clinical assessment and cranial computed tomography (CT) 24-48 hours before and 3 months after CEA. S100b serum levels were evaluated before surgery, at cross-clamping, 10 minutes later, at declamping, and 24-48 hours and 10-12 weeks after CEA. Increased S100b levels were detected in 11 patients (39%); eight (73%) of these patients had symptomatic carotid artery disease. Increased S100b level correlated with history of TIA or stroke ( p=0.005), low mini-mental state examination score ( p=0.02), and ischemic infarctions at preoperative CT ( p=0.03). Slight and transient increased S100b levels were detected in 39% of patients during CEA. The protein levels increased despite the absence of clinical events during surgery. Our findings suggest a failure of compensatory hemodynamic or metabolic mechanisms in peri-ischemic tissue, whose longterm effects on cognition remain to be investigated.

Plasma and tissue prolactin detection in colon carcinoma.

Serum concentrations of prolactin, a trophic hormone produced by the pituitary gland, have been shown to be raised in certain group of patients with cancer. Prolactin was detected in 0-20% of the colon cancer by immunohistochemistry and in plasma in 6-53% of the patients. These conflicting results do not support the hypothesis of an ectopic prolactin production by colon carcinoma. The aim of this study was to confirm the reported incidence of hyper-prolactinemia in colorectal cancer and to find further evidence for an ectopic prolactin production by the tumor. Thirty consecutive patients with colon carcinoma were studied. Before surgery all the patients underwent blood sample collection to assay plasma prolactin levels. All patients underwent colectomy. All the neoplastic specimens were tested with antiprolactin antibody. In none of the patients were significantly high preoperative levels of plasma prolactin found. Prolactin immunostaining was not identified in any of the tumor specimens. We could not confirm previous reports of frequent hyperprolactinemia in patients with cancer. This is the first report in which the incidence of both hyperprolactinemia and prolactin positive immunostaining was 0%. Our study was unable to demonstrate the synthesis of prolactin by colorectal cancers. The tumor is unlikely to be the source of hormone production. Our results suggest that circulating prolactin levels cannot be used as prognostic marker in patients with colon cancer.

Plasma levels of endothelin-1 increase in patients with sarcoidosis and fall after disease remission.

BACKGROUND: Sarcoidosis is a chronic systemic disease, characterized by an imbalance of immunity processes and the presence of granuloma. Endothelin-1, a new vasoactive and bronchoconstrictive peptide, is a powerful mitogen for smooth muscle cells and fibroblasts and plays a role in the inflammation state. We postulate that endothelin-1 has a role in sarcoidosis. METHODS: We studied the behaviour of circulating levels of endothelin-1 in 20 patients with sarcoidosis and its correlation with some biochemical parameters of activity disease, such as erythrocyte sedimentation rate (ESR) and serum angiotensin-converting enzyme (SACE). We measured serum levels of ESR, SACE, calcium and plasma endothelin-1 levels in all patients at the beginning of the study and one again in 9 patients with clinical-biochemical remission of disease after steroid treatment. RESULTS: In patients with sarcoidosis, circulating levels of endothelin-1, SACE and ESR were significantly higher (p<0.001) than those of healthy subjects. Moreover, in patients with pulmonary involvement, there was a significant statistical difference (p<0.001) between endothelin-1 levels and radiological stage compared to normal subjects. In the 9 patients with remission of disease, both endothelin-1 levels and parameters of activity disease normalized. CONCLUSIONS: Our results seem to suggest that the increase of plasma endothelin-1 levels in active sarcoidosis can represent an expression of the endothelial dysfunction and reflect the picture of cellular activation.

A case of pheochromocytoma with renal artery stenosis and post-surgical watery diarrhea.

A 35-year-old woman was admitted to our hospital with the following complaints, headache, sweating, anxiety, dizziness, nausea, vomiting and severe hypertension. The technical images (abdominal CT, scintigraphic octreotide scan and renal arteriography) revealed the presence of a left adrenal pheochromocytoma and stenosis of the renal artery. Ten days following adrenalectomy, watery diarrhea appeared. The long-acting somatostatin analogue octreotide (LAR, 30 mg/month, i.m.), was started, and after 2 weeks diarrhea decreased and gradually disappeared. In conclusion, we were confronted with an unusual case of pheochromocytoma associated with renal artery stenosis and the appearance of watery diarrhea some days after surgical treatment. Treatment with octreotide brought about the remission of diarrhea in this patient.

[Incidentalomas of the adrenal glands. Personal cases and review of the literature]. / Incidentalomi del surrene. Casistica personale e revisione della letteratura.

BACKGROUND: The growing use of sensitive non-invasive methods to image the abdomen such as ultrasonography, computed tomography (CT) and magnetic resonance imaging (MRI), have resulted in increasing recognition of adrenal incidentalomas. METHODS: In this study, we report the clinical, endocrine and radiologic evaluation of 75 patients (50 women and 25 men, mean age 56 +/- 14 years) with adrenal incidentaloma (size 1 to 18 cm, mean 3 +/- 2.5 cm). None of the patients showed any symptoms or clinical signs that might indicate the existence of adrenal dysfunction. The patients underwent basal and dynamic evaluation of the hypothalamic-pituitary-adrenal axis, renin-angiotensin-aldosterone system, and adrenomedullary function. Moreover, CT and MRI scan and Iodo-cholesterol (NP-59) scintigraphy were performed. RESULTS: The endocrine evaluation indicated 11 cases of pre-clinical Cushing's syndrome, 2 cases of pheochromocytoma and 62 not-functionally adrenal masses. On the basis of endocrine and morphologic data, 29 patients underwent surgical treatment: 20 adrenocortical adenoma, 2 pheochromocytomas, 2 not-cortisol-secreting adrenal carcinomas, 1 adrenal lymphoma, 1 adrenal metastasis, 1 myelolipoma, 1 hemorrhage and 1 pseudocystis. CONCLUSIONS: In conclusion, patient with an incidentally discovered mass has to be investigated to detect malignancy and subtle hormonal overproduction, to select the cases for surgical treatment.

Cryptic epitopes on alpha-fetoprotein induce spontaneous immune responses in hepatocellular carcinoma, liver cirrhosis, and chronic hepatitis patients.

To determine alpha-fetoprotein (AFP) immunogenicity in vivo, the presence of antibodies in sera of 60 hepatocellular carcinoma, 15 liver cirrhosis, and 15 chronic hepatitis patients was evaluated by Western blotting and immunoprecipitation analyses using purified human AFP. High titers of anti-AFP immunoglobulins were detected in 14 hepatocellular carcinomas (P = 0.0006), 3 liver cirrhosis (P = 0.0173), and 1 chronic hepatitis patient, but they were not detected in 40 healthy individuals. Therefore, spontaneous immune responses to AFP are significantly associated to liver diseases (P = 0.0015). Patient immunoglobulins recognized proteic linear epitopes that were cryptic in the native protein, as demonstrated by their restricted reactivity with denatured deglycosylated AFP. Thus, in pathological liver conditions, tolerance to this self-molecule is circumvented. The identification of AFP immunogenic epitopes may contribute to defining novel immunotherapeutic strategies targeting this antigen.

[The intracystic concentration of MCA in the fluid from large breast cysts]. / Concentrazione intracistica dell'MCA nel fluido di grosse cisti della mammella.

75 patients with breast gross cystic disease and no cancer have been included in the study. For each patients serous and intracystic concentrations of MCA have been measured. The aim of the study is to assess whether if a relation between intracystic concentration of the marker and resistance and capability of cellular reproduction exists (confirmed by the release of the cyst). The analysis of intracystic values shows that synthesis of MCA is an intrinsic peculiarity of cytologic kind. It is apparently independent from inflammatory or hemorrhagic contemporary processes.

Comparison of CEA, MCA, CA 15-3 and CA 27-29 in follow-up and monitoring therapeutic response in breast cancer patients.

In order to define the most useful tumor marker panel in breast cancer patients' follow-up and in monitoring treatment response, serological levels of CEA, MCA, Ca 15-3 and Ca 27-29 were evaluated in 220 patients. 180 patients had no evidence of disease (NED) after primary treatment, and 40 had metastases at first diagnosis time; in a 4 years follow-up, 30 of the NED patients relapsed, and were then included in the group of metastatic patients subjected to anticancer treatment. Overall sensitivity in metastatic patients was: CEA 40%, MCA 35%, Ca 15-3 79%, Ca 27-29 70%, with the highest percentages and mean values in liver and bone localizations. Combination of Ca 15-3 and Ca 27-29 improved sensitivity in bone lesion (85% vs 80%), in locoregional relapses only association with CEA increased sensitivity (60% vs 40%). Ca 15-3 and Ca 27-29 values increased on average 3 months before clinical diagnosis. In treated patients there was a better correlation with a clinical course of disease for Ca 15-3 and Ca 27-29 (both 81%) as compared to the other determined markers.

[Production of PSA by breast adenocarcinoma cells: preliminary results]. / Produzione del PSA da parte di cellule di adenocarcinoma della mammella: risultati preliminari.

PSA has been measured out with highly sensitive method both in liquid of mammary cysts and in cytosol of neoplastic cells. Our data point out that dysplastic, metaplastic and anaplastic breast epithelial cells produce PSA. Such production is, probably, related to expression of hormonal receptors.

[Immunoradiometric assay of Ca 125 in breast cystic fluid]. / Dosaggio immunoradiometrico del Ca 125 nel liquido di grosse cisti della mammella.

Ca 125 was assayed in serum and in breast cyst fluid of 78 patients with GCD. Levels of the marker in cyst fluid are generally < 30 U/ml. More significative results were found in relation to cysts relapse. In apocrine cysts relapse levels were higher than 270 U/ml, in serum cysts not relapsed levels were between 31-270 U/ml. Ca 125 could have a significative role in cell differentiation and its control.

CEA, TPA, CA 19-9, SCC and CYFRA at diagnosis and in the follow-up of anal canal tumors.

The authors evaluated serum CEA, TPA, CA19-9, SCC and CYFRA at diagnosis and in the follow-up of 18 anal canal tumors. Sensitivity at diagnosis was 38.8% for CEA, 55.5% for TPA, 16.6% for CA19-9, 50% for SCC and 5.5% for CYFRA. In the follow-up CEA showed 0% sensitivity and 73.3% specificity, TPA 33.3% sensitivity and 86.6% specificity, CA19-9 0% sensitivity and 80% specificity, SCC 0% sensitivity and 93.3% specificity, CYFRA 0% sensitivity and 100% specificity. The authors consider the usefulness of serum SCC and TPA at diagnosis of squamocellular anal cancer and of CEA in the diagnosis of cloacogenic tumors. Nevertheless these serum markers did not detect recurrences in the follow-up.

[Levels of Cathepsin D in breast cyst fluid]. / Dosaggio della Catepsina-D nel fluido di macrocisti della mammella.

Cathepsin-D was assayed in serum and in breast cyst fluid of 60 non neoplastic patients with GCD. The results are independent from cytological type or possible cyst relapse. Although the study confirms the high levels of Cat-D in breast cyst fluid no predictive value has been demonstrated. Its expression may be related to systemic endocrine factors.

[Level of immunosuppressive acidic protein in the fluid of breast cysts]. / Dosaggio della proteina acida immunosoppressiva nel liquido di cisti mammarie.

IAP marker was detected by radio-immunodiffusion in breast gross cyst fluid and in serum. No correlation was found between this antigen and cyst' cytological type, serum concentrations and acute phlogosis. In most cysts IAP was not valuable; on the other hand, high levels of glycoprotein were found only in the subgroup on relapsed class of apocrine cysts.

Serum ostase in the follow-up of breast cancer patients.

The present study was carried out on 152 patients divided into three groups: A) 73 underwent radical surgery for breast carcinoma without signs of metastases; B) 31 patients with radiologic and scintigraphic evidence of bone metastases originating from malignant mammary neoplasia (14 with only one and 17 with two or more localizations); C) 48 affected by simple mammary cysts. No patients had a previous history of primary or secondary bone pathologies or renal, hepatic or endocrine ones. Besides this, no patient took drugs influencing the metabolic turnover of the bony tissue in the three months preceding the study. After surgery all patients underwent standard clinical and laboratory follow-up, the latter including, every 3 months, the evaluation of serum CA 15.3, CA 27.29 MCA, and ostase. The ostase cut-off, obtained by the statistical elaboration of the serum values of the 48 patients with benign mammary cysts and the 73 disease free patients, was 17 microg./L. The mean concentration in the three groups and two subgroups was: 13.76 microg./L (patients without metastases), 31.84 (patients with metastases), 18.4 (limited bony metastases), 40.04 (diffused bony metastases) and 5.36 (mammary cists). The diagnostic sensitivity of ostase proved superior to that of CA 15.3 (84% vs 75%) except when considering the subgroup with limited metastases (71.4% vs 72.7%), while the specificity was similar (around 78%). CA 27.29 and MCA were not useful as markers of metastasis. In a longitudinal-perspective study it was possible periodically to test these markers in 13 patients, at first, disease free and then with signs of bone progression evidence by skeletal scintigraphy. In 11 of these patients ostase and CA 15.3 showed increased values, an average 136 and 131 days respectively, before instrumental evidence of progression. None of the 13 patients, at the time of bone progression diagnosis, showed clinical, laboratory or instrumental signs of disease in other organs. The precocity of the serum increase of ostase could have a triple role: 1) accomplishment of a closer follow-up in patients at "high risk" of bone disease; 2) aid in the interpretation "in a neoplastic sense" of an "uncertain image of hypercaptation"; 3) accomplishment of a supporting or specific oncology treatment at an earlier stage which may be of some advantage as regards quality of life.

Skeletal alkaline phosphatase as a serum marker of bone metastases in the follow-up of patients with breast cancer.

Immunoradiometric determination of the bone isoenzyme of alkaline phosphatase with a method provided by Hybritech Inc., San Diego CA (USA) was carried out in 145 female patients, 97 of whom with radically operated breast cancer and 48 with benign mammary cysts, in order to evaluate the correlation of serum levels with the metabolic process of bone rearrangement in patients with bone metastases. This study shows that skeletal ALP, having high specificity (86.48%) and sensitivity (78.6%) for early progression (the average anticipation time compared to scintigraphic detection was 101 days) could represent a valid marker for bone metastases in association with mucinous markers in the follow-up of patients operated for breast cancer. In addition, dynamic serum determination of skeletal ALP could be a valid help in monitoring the efficacy of therapy in patients with bone progression.

[The prognostic significance of the measurement of Ca 15-3 in the fluid of large breast cysts]. / Significato prognostico del dosaggio del Ca 15-3 nel liquido di grosse cisti della mammella.

Ca 15-3 was assayed in serum and in breast cysts fluid of 78 non neoplastic patients presenting Gross Cysts (GCD). In apocrine cysts and in the mixed type (serous and apocrine, class III) relapse was indicated by high serum and intracystic levels of the marker. Ca 15-3, therefore, may discriminate in a group of cysts a higher cellular resistance as well as an increased cell proliferation. Results suggest an important role of the marker in the follow up of patients with GCD and for the early detection of cyst relapse.