Davaris P

Heat shock protein 70 and HLA-DR molecules tissue expression. Prognostic implications in colorectal cancer.

PURPOSE: The expression of 70,000-Da heat shock protein (HSP 70) and HLA-DR molecules on cancer cells influences immunologic mechanisms that may be of some prognostic significance. The purpose of this study was to examine the relationship among immunohistochemical HSP 70, HLA-DR expression, and clinicopathologic tumor variables, as well as patient survival in a series of 128 colorectal carcinomas. METHOD: A three-step immunoperoxidase staining technique was undertaken for detection of both markers. RESULTS: Of the examined carcinomas 77.3 percent were HSP 70-positive and 74.2 percent were HLA-DR-positive. Increased HSP 70-positive expression correlated significantly with low differentiation (P<0.05), showed a tendency to characterize advanced stages of disease, and was clearly associated with worse overall survival (P<0.05). The highest rate of HLA-DR positivity was demonstrated in early stages and was significantly associated with more favorable prognosis (P<0.001). HSP 70-positive/HLA-DR-negative patients had worse overall survival compared with the rest (P<0.001). CONCLUSIONS: The resulting opposite effects on prognosis of examined markers seem to be related to different pathophysiologic functional roles on tumor immunology.

Immunohistochemical expression of C-myc oncogene, heat shock protein 70 and HLA-DR molecules in malignant cutaneous melanoma

The clinical course of malignant melanomas is frequently unpredictable, although a number of prognostically useful variables can be identified. There is a need for additional markers of prognostic value. In a series of 60 malignant cutaneous melanomas, we analysed the immunohistochemical expression of c-myc proto-oncogene, heat shock protein 70 (HSP70) and HLA-DR molecules in order to investigate their prognostic significance. C-myc, HSP70 and HLA-DR were expressed in 43.3%, 56.6% and 38.3% of all melanoma cases, respectively. Advanced Clark levels (Clark III–V) were significantly associated with c-myc expression rate (P<0.05), HSP70 detection (P<0.01) and HLA-DR positivity (P<0.01). Increased Breslow thickness (>1.5 mm) was related to HLA-DR expression (P<0.05). High mitotic rate was closely associated with c-myc positivity (P<0.05), while HSP70 and HLA-DR expression separately correlated to clinical stage of the disease (P<0.05). The evaluation of these variables may be of immunological and prognostic significance. They were found to be associated with melanocyte subpopulations of the vertical growth phase which are arguably characterized by an increased invasive potential.

Prognostic implications of aberrations in p16/pRb pathway in urothelial bladder carcinomas: a multivariate analysis including p53 expression and proliferation markers.

Objective: To assess the prognostic value of the expression of two negative regulators of the cell cycle, namely CDKN2/INK4a gene product (p16) and retinoblastoma gene product (pRb), in urinary bladder cancer in relation to clinicopathological parameters, proliferative fraction and p53 protein accumulation. Methods: Paraffin sections from 139 patients with urothelial carcinomas were stained immunohistochemically with antibodies to p16 (F12), pRb (PMG3–245), p53 (DO1), PCNA (PC10) and Ki–67 (MIB–1). Results: Diminished p16 and pRb expression occurred in 29 and 74% of cases, respectively, being associated with advanced stage but not with histological grade, papillary status or proliferation rate. In most cases (53%) with some fault in the p16/pRb pathway, only one gene was affected. A double–negative p16/pRb phenotype was comparatively uncommon (25%) and was usually seen in T3–T4 tumours. In survival analysis (either univariate or multivariate) aberrant p16 expression was an adverse prognostic parameter only in T3–T4 tumours. In contrast, the abnormal p16/pRb and p53/p16 phenotypes were linked to a diminished overall and disease–free survival (univariate analysis); p53/p16 abnormal expression was also found to be an independent predictor of reduced survival in muscle–invasive tumours, while proliferation markers were the only parameters with independent significance in superficial (Ta–T1) tumours. Conclusion: Our results suggest that lack of p16 immunoexpression, when combined with p53 accumulation, plays an important role in determining the clinical outcome in muscle–invasive urothelial carcinomas.

Mitosin and DNA topoisomerase IIalpha: two novel proliferation markers in the prognostication of diffuse astrocytoma patient survival.

The expression of two novel proliferation-associated markers, mitosin and topoisomerase II&agr; (Topo II&agr;), was evaluated immunohistochemically in consecutive paraffin sections from 60 diffuse astrocytomas (grades 2 to 4) in relation to clinicopathologic parameters, proliferating cell nuclear antigen (PCNA) and Ki-67 (MIB-1) expression and survival. The percentage of mitosin and Topo II&agr;–positive cells (LI) increased with grade and Ki-67 LI, but could not discriminate between grade 3 on the one hand and grades 2 or 4 on the other hand. In 51% of cases, Ki-67 LI exceeded Topo II&agr; LI, especially within grade 4. Topo II&agr; and mitosin expression was adversely related to overall and disease-free survival in the entire cohort and in grades 2/3. However, only Topo II&agr; LI affected disease-free survival in grade 4 tumors. Multivariate analysis selected only mitosin LI along with the age of the patient, as the independent parameters predicting overall survival, whereas Topo II&agr; emerged as the single independent predictor of disease-free survival. It is concluded that the proliferative potential of astrocytomas, as measured by mitosin and Topo II&agr; immunostaining, conveys useful prognostic information, in addition to that obtained by standard clinicopathologic parameters.

Immunodetection of gastrin-releasing peptide in malignant melanoma cells.

Gastrin-releasing peptide (GRP), the mammalian counterpart of bombesin, was first identified in the nervous system of the gastrointestinal tract. Little is known about its distribution in the human skin or about its function in certain diseases such as malignant melanoma. Recently functional GRP receptors have been found on human melanoma cell lines. We therefore investigated, using immunohistochemistry, whether human melanoma cells express GRP and whether there is a significant change in its distribution among the different clinical types of melanoma and a connection to histopathological features such as growth phase, type of malignant cells, Breslow thickness and Clark level of invasion. We demonstrated the existence of GRP in all clinicopathological types of melanoma; a predilection for quantitatively increased GRP immunostaining was noticed in nodular melanomas (P= 0.007). As well as this, we observed a restriction of GRP expression at a specific level of invasion, i.e. within the reticular dermis (Clark IV) (P= 0.032). GRP immunoreactivity was found to be associated with an increased amount of melanin pigment in malignant cells (P= 0.054). The presence of GRP in malignant melanocytes, along with its association with the various histopathological features, suggests that GRP may play a role in the pathophysiology of this type of cutaneous tumour.

Membranous glomerulonephritis: a morphometric study.

Archival material from 45 renal biopsies with a diagnosis of idiopathic membranous glomerulonephritis (MGN) were studied by computer-aided image analysis in order to evaluate the prognostic significance of glomerular and interstitial morphometry in MGN. The control group consisted of thirty seven normal renal biopsy specimens. The surface area, the perimeter, the major axis length and the shape factor of renal glomeruli as well as the percentage of the interstitial fibrosis were measured. All the morphometric parameters related to the size of glomeruli had significantly higher values in the patient group (p = 0.000 for all the parameters). However, no significant difference of the glomerular size between different stages of MGN was observed. In contrast, the percentage of interstitial fibrosis increased as the MGN stage rose (median values: 10.3% in stage 1, 14.2% in stage II, 26.9% in stage III, 28.9% in stage IV and 34.2% in stage V, Kruskal-Wallis ANOVA H = 37.645, p = 0.000). In the multivariate analysis the percentage of interstitial fibrosis was the only independent prognostic factor (p = 0.013). Our findings suggest that, in membraneous glomerulonephritis, the interstitial fibrosis increases as the MGN stage progresses, while the size of renal glomeruli has increased at a very early stage of the disease. This fact may indicate that interstitial fibrosis, not glomerular lesions, is mainly responsible for the reduction of renal function.

Effects of submucosal Teflon paste injection in vesicoureteric reflux: results with 1- and 2-year follow-up data.

One hundred refluxing ureters with grades II (n = 65), III (n = 27) and IV (n = 8) vesicoureteric reflux (VUR) were treated with endoscopic submucosal polytetrafluoroethylene (Teflon) paste injection. Reflux stopped in 77 ureters after a single injection and in 13 ureters following a second injection. Two ureters remained with minimal (grade I) residual reflux, whilst in another 5 improvement from grades III and IV to grade II reflux was noted. In 3 ureters reflux remained unchanged, because of lateral ectopia of the refluxing ureteric ostium, hindering the injection of Teflon into the correct position. Re-evaluation at 1 year postinjection, of 48 previously cured ureters showed recurrence of reflux in 1 (2%), but no further recurrences were noted in 19 ureters reevaluated after 2 years.

Cecum malakoplakia: A tumor-like lesion with coexistent adenocarcinoma

We report the case of a 75-year-old Caucasian male who presented with acute abdomen and fecal leakage from his old appendectomy scar and required exploratory laparotomy. A large cecal mass was found and a right colectomy was performed. At pathology, the neoplastic mass was identified as malakoplakia with a small area corresponding to a moderately differentiated colonic adenocarcinoma.Occurrence of malakoplakia in the cecum, associated with adenocarcinoma, is extremely rare if we take into account the limited number of the reported cases of its coexistence with colonic cancer; our case is the second report of such an entity in the cecum. The unusual presence of fistula to the appendectomy scar may be related to the infiltrative nature of the histiocytes constituting this process. Immunochemical studies can assist in the histopathologic differentiation of malakoplakia from other entities that might represent with this tumor-like configuration.

CARCINOGENESIS ASSOCIATED WITH DIMETHYL SULFOXIDE

Panayotis Davaris, Laboratory of Morbid Anatomy and Histopathology, Athens University Medical School, GR-115 27 Athens (Greece) Dear Sir, We wish to draw your attention to the possible association between dimethyl sulfoxide (DMSO) and carcinogenesis. During an experimental project on bladder cancer, dibutyl-n-nitrosamine was administered to white Wistar rats subcutaneously diluted in DMSO. The daily dose of the carcinogen ranged from 30 to 100 mg/kg, and the exposure lasted from 50 to 320 days. We noted that apart from the appearance of transitional cell bladder carcinomas, one third ofthe animals also developed lung tumors. These tumors appeared after at least 150 days of exposure and were adenocarcinomas originating from the alveolar epithelium. This finding is compatible with previous observations made by Sanders et al. [1], who reported the induction of cancer in the lung of rats given dibutyl-n-nitrosamine subcutaneously with DMSO as its solvent in 1974. These authors postulated that DMSO could have an organo-tropic action, directing the carcinogenic effect of dibutyl-n-nitrosamine upon the lung. They also noted that dibutyl-nnitrosamine diluted in cellulose oil failed to cause any tumors in the lung in another series of animals. DMSO is a potent organic solvent widely used in industry [2]. Since 1968 it has been used in urology for the treatment of patients with interstitial cystitis in the form of intravesical instillations [3]. Its therapeutic effect seems to be achieved by means of local chemical denervation [4]. Although to date no serious side effects have been reported from such treatment, the possibility of carcinogenesis associated with DMSO should be given consideration before advocating its administration.

Synchronous Comparative Evaluation of Fibronectin Continuity and Cellular Growth Kinetics in Malignant Cutaneous Melanoma

Cellular fibronectin mediates the adhesion of cells with the extracellular matrix and thus probably regulates tumor invasion. Evaluation of proliferating cell nuclear antigen expression and counting of silver nuclear organizer regions potentially assess proliferative fraction and biologic aggressiveness in several malignancies. We investigated the above markers’ prognostic value in 60 malignant melanomas of the skin. Deep Clark levels were statistically associated with discontinuous fibronectin staining pattern (SPII) (P < .05) as well as with high proliferating cell nuclear antigen and silver nuclear organizer regions indexes (P < .01 and P < .05, respectively). Fibronectin staining pattern (SPII) was frequently observed in thick melanomas (P < .05). High proliferating cell nuclear antigen immunoreactivity positively correlated with mitotic activity (P < .05), the presence of vertical growth phase (P < .05), and increased silver nuclear organizer regions counts (P < .05). Patients with continuous fibronectin staining pattern (SPI) and low proliferating cell nuclear antigen scores were unlikely to develop metastatic disease (P < .01). Therefore, the simultaneous comparative assessment of these factors is very promising in predicting melanomas clinical evolution.