Evaluation of the prognostic and predictive value of p53 and Bcl-2 in breast cancer patients participating in a randomized study with dose-dense sequential adjuvant chemotherapy.

PURPOSE: To assess the prognostic and predictive significance of p53 and Bcl-2 protein expression in high risk patients with breast cancer treated with dose-dense sequential chemotherapy. PATIENTS AND METHODS: From June 1997 until November 2000, 595 patients were randomized to three cycles of epirubicin (E) 110 mg/m2 followed by three cycles of paclitaxel (P) 250 mg/m2 followed by three cycles of 'intensified' CMF (cyclophosphamide 840 mg/m2, methotrexate 47 mg/m2 and fluorouracil 840 mg/m2) or to four cycles of E, followed by four cycles of CMF. p53 and Bcl-2 expression was investigated by immunohistochemistry in 392 and 397 patients respectively. RESULTS: Positive expression of p53 was detected in 104 (26.5%) patients and was significantly associated with negative hormonal status, worse histologic grade, higher incidence of disease relapse and higher rate of death. p53 positive expression was a significant negative predictor of overall survival (OS) (P = 0.002) and disease-free survival (DFS) (P = 0.001). Negative expression of Bcl-2 was detected in 203 (51%) patients and was significantly associated with negative hormonal status. Multivariate analysis revealed that, positive p53 expression, higher number of positive nodes and worse tumor grade were related to significantly poorer OS and DFS. CONCLUSIONS: For both treatments, p53 positive expression was a significant negative prognostic factor for OS and DFS while Bcl-2 was not. No predictive ability of p53 status or Bcl-2 status for paclitaxel treatment was evident.

u-PA expression in benign, borderline and malignant ovarian tumors.

The malignant potential of solid tumors is related to their ability to invade adjacent tissue and to metastasize. The plasminogen activation system is one of the critical factors in tumor progression since it is involved in tumor invasion and metastasis. This study was performed to examine the expression of u-PA in benign, borderline and malignant tumors of the ovary by immunohistochemical evaluation on formalin-fixed, paraffin-embedded specimens applying monoclonal antibody 3689 directed to the b-chain of u-PA. Normal epithelial cells of the ovary (n = 5) showed no staining of u-PA but some stromal cells were slightly stained. Invasive carcinomas (n = 16) and borderline tumors (n = 15) showed a moderate to strong diffuse cytoplasmic staining. Benign tumors (n = 20) showed a variety of staining. The observation of randomly positive u-PA stromal cells is noteworthy. The percentage of u-PA-positive tumors was higher in carcinomas than in other tumors. There was no correlation with other known risk factors of malignancy such as differentiation, stage or type of tumor. In conclusion there are noticeable differences in u-PA expression among ovarian tumors and u-PA increase in ovarian tumors can be attributed to an increased diffuse cytoplasmic content in the neoplastic epithelial cells.

Malignant lymphoma in nasal cavity and paranasal sinuses of a dog.

A case of a canine large cell type T-cell lymphoma, with features of high-grade malignancy is described. The tumour was found confined in the nasal cavity and the paranasal sinuses of a crossbred German Shepherd dog. Histological examination revealed the features of a highly malignant large cell lymphoma. Ultrastructurally, the lymphoid tumour cells bore cytoplasmic protrusions that interdigitated tightly. From a panel of tumour markers used, the neoplastic cells were stained only for vimentin. Immunophenotyping of the tumour cells by means of CD3, CD79, kappa-light chains and lambda-light chains detection was undertaken. The tumour stained only for CD3 and was classified as T-cell lymphoma.

Effect of navelbine on inhibition of tumor growth, cellular differentiation and estrogen receptor status on Lewis lung carcinoma.

We studied the antitumor activity of Navelbine (NVB) together with its ability to induce cellular differentiation and to influence estrogen receptor status of Lewis lung carcinoma (LLC). A total of 32 C(57)B1 mice divided into 5 groups were used for transplantation of LLC. Four groups of mice were treated with 5.0, 2.5 and 1.25 mg/kg/day from day 1 to 9 and 1.25 mg/kg on days 1, 7 and 13. Eight mice were controls. The dose of 1.25 mg/kg/day was the most effective and produced 72.7% inhibition of tumor growth. Ultrastructurally, on day 1 the cells showed poor differentiation. On day 14, in the case of 72.7% inhibition of tumor growth the study revealed a significant restoration of the morphology of the cells. Estrogen receptors gave a positive value in contrast to the initial measurement which was negative. The present study demonstrated good antitumor activity of NVB on LLC. NVB may also induce cellular differentiation and influence the estrogen receptor status.

A new approach in the management of urothelial tumors using GM-CSF on marker lesions: an ultrastructural and immunohistochemical study on the macrophage population in bladder mucosa.

Our purpose was to investigate a new therapeutic model, GM-CSF-targeted immunomodulation on transitional cell carcinoma (TCC) marker lesions and to evaluate the immunologic response of the bladder mucosa. Eleven patients with pTa or pT1 bladder cancer were eligible for the study. All lesions were removed by transurethral resection (TUR) except for a marker lesion. All patients received 8 weekly instillations of 300 microg of GM-CSF, after which cystoscopy with bladder biopsies +/- TUR was repeated on adjacent urothelium or tumor or both. Paraffin-embedded sections were immunohistochemically stained with CD68, which labels monocytes and macrophages. The CD68+ cell population was evaluated as 1+ to 3+. Comparable specimens were routinely processed for ultrastructural analysis. Complete response was observed in 6 patients (55%), persistent tumor occurred in 4 patients (approximately 36.4%), and 1 patient (8.6%) showed recurrence. Immunohistochemically, an at least twofold increase in the number of the CD68+ cells was observed in all responders. Submicroscopically, migration of macrophages to the surface layer occurred. Macrophages showed an extensive lysosomal system and pseudopodia. This study indicates that the prophylactic treatment of TCC with GM-CSF may induce immunomodulatory effects on macrophage activities, which could be associated with the clinical evolution of the disease.

Natural killer (NK) cell assay within bladder mucosa of patients bearing transitional cell carcinoma (TCC) after interferon (IFN) therapy: an immunohistochemical and ultrastructural study.

The authors studied the number and the ultrastructural evidence of NK cell activation in the non-involved urothelium in patients with transitional cell carcinoma (TCC) of the urinary bladder, before and after transurethral resection (TUR) and interferon (IFN) therapy. Eight male patients, free of recurrence 1 year after TUR and IFN-a2b therapy, were studied. Each patient received 22 instillations of 50MU of IFN-a2b over a period of 1 year. Two specimens from the non-involved urothelium, one adjacent to and another away from the tumour, were obtained before and after therapy, for immunohistochemical and ultrastructural studies. The number of NK cells was evaluated immunohistochemically in paraffin sections with the CD57 monoclonal antibody, and their activation was detected by routine electron microscopy processing. Before treatment, few NK cells were randomly found in the lamina propria. At the end of therapy, however, their number increased and NK cells were found to infiltrate the urothelium, a finding that was not observed before treatment. The number of NK cells did not correlate with the degree of the inflammatory infiltrate of the mucosa. Moreover, the ultrastructural study revealed activation of NK cells with enhanced cytolytic activity. IFN therapy increases the number and promotes the activation of NK cells within the bladder mucosa. This finding could be of clinical significance in the prevention of tumour recurrence, given that NK cells enhance the immunological defense mechanisms of the bladder.

Correlation of apoptosis with the distribution and composition of lymphocytic infiltrate in human breast carcinomas.

We studied the lymphocytic infiltrate in human breast carcinomas and correlated its distribution and composition with the apoptotic index of cancerous cells. Paraffin sections from 36 human breast carcinomas with prominent lymphocytic infiltration were immunohistochemically stained with L26, UCHL1 and Leu-7 antibodies. Apoptotic index was expressed as the number of cells undergoing apoptosis per 20 HPF. The lymphocytic infiltrate was composed mainly of T-cells, usually peripherally but in 17 cases intratumorally distributed. T-cells, in contact with apoptotic cells, were observed in 11 cases. B-cells formed small aggregates at the periphery of the tumor. NK cells were present isolated in the infiltrate. No correlation was found between the type and distribution of the lymphocytic infiltrate and apoptotic index. The presence of intratumoral T-lymphocytes, occasionally observed in contact with apoptotic cells, may suggest that they could be partly associated with the apoptotic death of cancerous cells.

Ultrastructural study of the urothelial lysosomal system in patients with transitional cell carcinoma after transurethral resection and interferon therapy.

The epithelial cells of human bladder urothelium contain a prominent lysosomal system on the surface layer which involves autophagic, phagocytotic and excretory processes. The noninvolved urothelium of tumor-bearing patients, however, does not contain this well-developed lysosomal system. Interferon restores the differentiation of the urothelium. Its action on the lysosomal system, however, has not been investigated. We studied ultrastructurally the noninvolved urothelium of eight patients with transitional cell carcinoma who after transurethral resection and intravesicular interferon instillations for 2 years did not develop recurrence. We noted that the number and size of lysosomes increased, being most numerous within the cells of the surface layer. Characteristic large lysosomes with the morphology of multivesicular bodies were also evident. These multivesicular bodies were almost entirely filled with small vesicles containing a dense core. Our findings show that after 2 years of interferon administration a re-appearance of a highly developed lysosomal system of the noninvolved urothelium was evident. This restoration to the normal morphology with reappearance of the lysosomal system, which could be partly attributed to interferon therapy, may be of clinical significance for prevention of tumor recurrence.

Natural killer cell activation after interferon administration in patients with metastatic renal cell carcinoma: an ultrastructural and immunohistochemical study.

OBJECTIVE: We studied the intralesional number of natural killer (NK) cells and the ultrastructural evidence of their activation in patients with metastatic renal cell carcinoma, after preoperative interferon (IFN) administration. METHODS: Tumor sections from 10 patients with metastatic renal cell carcinoma who received preoperative IFN (5 MU x 3 days) and from 10 patients with renal cell carcinoma were obtained. The number of NK cells was estimated immunohistochemically in paraffin sections with the CD57 monoclonal antibody and their activation was evaluated ultrastructurally by routine electron microscopy processing. RESULTS: The number of NK cells was increased in the tumors of the IFN-administered patients (5.3 cells vs. 1.6 in the controls, p = 0.03) while their ultrastructural features revealed activation and enhanced cytolytic activity, through facilitation of granule content release. CONCLUSIONS: IFN promotes the aggregation and activation of NK cells within the renal cell tumor, further strengthening the concept of its immunomodulatory activity.

A new approach combining interferon and bromocriptine LA to prevent spontaneous mouse mammary carcinoma.

The development of spontaneous mammary carcinoma in C3H mice is mainly attributed to an oncogenic virus (MMTV) and the high prolactin levels of this inbred strain. Interferon (IFN) reduces the incidence of mammary carcinoma in virgin C3H mice when administered during lactation. Bromocriptine LA, an antiprolactin drug, also decreases the incidence of this tumor in C3H mice. The combined action of mouse IFN-alpha + beta and bromocriptine LA had a significant anticarcinogenic effect more significant than either of the above agents administered independently. This antitumor action was effective in virgin C3H mice only.

Tissue reaction to deep cuff of Tenckhoff's catheter and peritonitis.

To define the type of reaction around the deep cuff of Tenckhoff catheters and possible implications in peritonitis development, a histological evaluation of the tissue reaction around and into the deep cuff was done. Eight catheters, functioning for 2 to 40 months and removed for various reasons were studied. In all cases foreign body granulomata and fibrosis were found, with prominent giant cell formation. An inert birefrigent material, probably dacron particles, was found surrounded by giant cells in fibrotic regions and in some cases appearing as cytoplasmic inclusions. In 6 cases an inflammatory infiltration was observed, while in 4 with relapsing peritonitis, polymorphonuclears and microabscesses were also found. It can be suggested that the tissue reaction to deep cuff predisposes to local infections and might be a factor for relapsing peritonitis.

Effect of bromocriptine on the ovarian cycle of two inbred strains of mice.

Mice of the C3H/Sy (high incidence of spontaneous mammary cancer) and AKR/Sy (low incidence of spontaneous mammary cancer) inbred strains, which have different hormonal profiles, were injected daily with bromocriptine for 1 month. The treatment increased the duration of the ovarian cycle of the AKR/Sy mice, whereas that of the C3H/Sy mice was not affected. It is suggested that the effect of bromocriptine on the ovarian cycle depends on the concentrations of plasma progesterone reached in each strain of mouse.

Decrease in mammary tumour incidence in virgin C3H mice given interferon only while suckling.

A high proportion of females of the C3H strain of mice develop tumours of the mammary gland which are caused by mouse mammary tumour virus (MMTV) transmitted through the milk. We have examined whether administration of mouse interferon (IFN) to nursing mothers and/or their suckling offspring only during the period of nursing, can affect the incidence of tumours developing in these animals. In two separate experiments, animals receiving IFN by direct injection while suckling, and remaining virgin showed a marked and statistically significant decrease in tumour incidence. Mice receiving the same or a tenfold higher dose of IFN while lactating showed no such reduction in tumour incidence, even if they had also received IFN while suckling. The results suggest that IFN can affect the initial establishment of the MMTV infection in suckling mice sufficiently to delay tumour development provided the animals are not exposed to the hormonal stimulus of pregnancy and lactation.