Practical value of the new system for Maspin assessment, in colorectal cancer.

Maspin is a serine protease which belongs to the serpin family and seems to play an important role in inhibiting angiogenesis and tumor proliferation. The significance of its expression in colorectal cancer (CRC) has not been elucidated so far. In our study, we tried to identify, based on Maspin expression, four groups of CRC, with possible prognostic impact. In 121 CRC, we analyzed the Maspin expression in correlation with the clinico-pathological features, microsatellite status and other markers such as p53, bax, bcl-2, VEGF (Vascular Endothelial Growth Factor) and CD31. Based on the percentage and intensity of Maspin expression in the tumor cells, the cases were grouped in four classes: negative, with cytoplasmic predominance, nuclear predominated, and cases with mixed (cytoplasmic-nuclear) expression. 9% of the cases were negative, 44% presented cytoplasmic predominance, the nuclear predominance was revealed in 24% of the cases, and the other 23% of CRC having a mixed Maspin positivity. The cytoplasmic predominance was correlated with a better prognosis, p53 negativity, bax positivity, and lack of tumor budding. Forty percent of microsatellite instable (MSI) cases presented mixed expression, this pattern being also related to a lower angiogenesis. Nuclear predominance was associated with p53 positivity, the lowest survival rate and intense VEGF expression. In conclusion, CRC with cytoplasmic predominance and mixed Maspin expression seems to present better prognosis whereas nuclear predominance is connected with high aggressivity.

The differences between the endothelial area marked with CD31 and CD105 in colorectal carcinomas by computer-assisted morphometrical analysis.

UNLABELLED: The aim of our study was to compare the CD31 and CD105 endothelial area (EA) by computer-assisted morphometrical analysis in colorectal carcinomas (CRC). Two hundred and eleven surgical specimens with CRC were immunohistochemical analyzed with markers for angiogenesis CD31 and CD105 (Endoglin). We determined the area of endothelial cells occupied in the microscope field (EA). RESULTS: The median area was 6.93 +/- 4.25% for CD31, respectively 5.65 +/- 2.23% for CD105. In the majority of cases, the CD31 EA was higher than CD105 EA. In the cases with the predominance of mature vessels, and also in the cases after radiotherapy, the CD105 EA was higher than CD31 (5.69 +/- 2.49%, respectively 10.23 +/- 5.93%). In our study, we tried to describe the clinico-morphological features of these cases. CONCLUSIONS: The CD105 seems to be the best marker for study of neoangiogenesis in CRC. Sometime, CD105 marks the activated endothelium of preexistent mature vessels. The radiotherapy destroys the neoformed but also the preexistent vessels. For the antiangiogenic treatment, it is important to determine the intensity of angiogenesis but also the type of neoformed vessels.

The angiogenesis in colorectal carcinomas with and without lymph node metastases.

UNLABELLED: Many clinical trials revealed that the anti-angiogenic treatment could improve prognosis in patients with metastatic colorectal carcinomas (CRC), when added to standard chemotherapy. In this paper, we tried to find out if the microvascular density (MVD) determined with CD31, CD105 was correlated with lymph node status, and if the intensity of angiogenesis was different in right versus left colon segments. We studied 187 CRC, with and without lymph node metastases, 128 from left and 59 from right colon. RESULTS: In the right colon, the MVD was higher in the cases where the lymph nodes did not present metastases (pN0) but also when four or more lymph nodes were involved (pN2). In the rectum and sigma, the angiogenesis presented the highest intensity in pN0 and pN1 stage (1-3 lymph nodes with metastases), decreasing in pN2 stage. In the descendent colon segment, the MVD did not present differences between the cases with and without lymph node metastases. CONCLUSIONS: Our study reveals that the most indicated cases for antiangiogenic treatment seem to be the pN0 and pN1 cases in the rectum and sigma, respectively pN0 and pN2 cases in the right colon. We tend to believe that the angiogenesis intensity in CRC is higher in early-stages of the tumoral proliferation but it is not an increasing process, having rather an oscillating character. Therefore, the angiogenesis remains an independent prognostic and predictive factor and the antiangiogenic treatment is necessary to be individualized for each patient.

Demonstration by in situ hybridization of ret proto-oncogene mRNA in developing placenta during mid-term of rat gestation.

Expression of the ret proto-oncogene (proto-ret) in rat conceptus tissues during development was examined by in situ hybridization using photobiotin-labeled oligodeoxyribonucleic acid probes corresponding to regions coding for the kinase and transmembrane domains of proto-ret gene product. High levels of the proto-ret transcripts were detected in the cytotrophoblasts in the placenta in the mid-gestational period (days 10 and 11), but on day 14 of gestation, when the placenta was undergoing morphological changes, transcripts could no longer be detected in the trophoblasts. These results suggest that the increased expression of proto-ret may be associated with the proliferation and/or differentiation of trophoblast cells at a specific stage. Improvements in the in situ hybridization technique by introduction of an ultrafast microwave energy fixation method, and repeated-reaction cycling of avidin-alkaline phosphatase and a biotinylated anti-avidin antibody for signal amplification, are also briefly discussed.

Effects of 4-week treatment with gastric carcinogens and enhancing agents on proliferation of gastric mucosa cells in rats.

Catechol and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) are gastric carcinogens in rats. Catechol, sodium chloride and bile salts have enhancing effects on gastric carcinogenesis induced by MNNG in rats. The effects of these compounds on proliferation of pyloric mucosa cells in male F344 rats were examined immunohistochemically using bromodeoxyuridine (BrdU) and anti-BrdU monoclonal antibody. Rats were given MNNG (83 micrograms/ml in their drinking water), catechol (0.8% in their diet), sodium taurocholate (0.3% in their diet), sodium taurodeoxycholate (0.3% in their diet), or sodium chloride (10% in their diet or by intragastric administration of 1 ml of saturated solution once a week) for 4 weeks. All these treatments markedly enhanced cell proliferation of the pyloric epithelium, suggesting the importance of enhanced cell proliferation in the development of gastric cancer.

Prognostic significance of residual disease after radiation therapy of stage III breast cancer.

239 Consecutive women with stage III carcinoma of the breast were treated by primary radiation therapy (RT) from 1977 to 1985. The response rate was 89%, the 5-year survival 40% and the local-regional tumor control (LTC) 59%. For the 27 non-responders, postirradiation chemotherapy was initiated but they died within 3 years. 105 Patients were subjected to mastectomy and axillary dissection after RT. In 107 cases, the RT was not followed by surgery. Systemic treatment consisted of hormonal therapy in both groups. The RT plus surgery group had better survival rate, 58% vs. 35% at 5 years. However, the incidence of less favorable cases was higher in the RT alone group. Histological findings in the operated group are analysed to determine prognostic significance of residual disease. No residual or only damaged microscopic disease was found in the breast in 36% of the cases. The axillary lymph nodes (ALN) were free of disease in 42% of the women. Patients with negative ALN after RT had significantly better 5-year survival (82% vs. 43%) and LTC (85% vs. 60%) rates. Decreased chest wall recurrence rate was associated with no residual or damaged microscopic disease in the breast (5% vs. 21%). The disease-free ALN were more common after 50-80 Gy telecobalt than after 40-50 Gy kV irradiation (51% vs. 33%). Primary tumor size (phi less than 5 cm vs. phi greater than 5 cm) had no significant impact on histological findings or on prognosis. Residual disease is mainly a marker of tumor-host relationship which indicates the biological aggressiveness of the disease.

Survival chance in papillary thyroid cancer in Hungary: individual survival probability estimation using the Markov method.

PURPOSE: The typically benign, but occasionally rapidly fatal clinical course of papillary thyroid cancer has raised the need for individual survival probability estimation, to tailor the treatment strategy exclusively to a given patient. MATERIALS AND METHODS: A retrospective study was performed on 400 papillary thyroid cancer patients with a median follow-up time of 7.1 years to establish a clinical database for uni- and multivariate analysis of the prognostic factors related to survival (Kaplan-Meier product limit method and Cox regression). For a more precise prognosis estimation, the effect of the most important clinical events were then investigated on the basis of a Markov renewal model. The basic concept of this approach is that each patient has an individual disease course which (besides the initial clinical categories) is affected by special events, e.g. internal covariates (local/regional/distant relapses). On the supposition that these events and the cause-specific death are influenced by the same biological processes, the parameters of transient survival probability characterizing the speed of the course of the disease for each clinical event and their sequence were determined. The individual survival curves for each patient were calculated by using these parameters and the independent significant clinical variables selected from multivariate studies, summation of which resulted in a mean cause-specific survival function valid for the entire group. On the basis of this Markov model, prediction of the cause-specific survival probability is possible for extrastudy cases, if it is supposed that the clinical events occur within new patients in the same manner and with the similar probability as within the study population. RESULTS: The patient's age, a distant metastasis at presentation, the extent of the surgical intervention, the primary tumor size and extent (pT), the external irradiation dosage and the degree of TSH suppression proved to be statistically significant and independent prognostic factors with regard to cause-specific survival in multivariate studies. During follow-up, 14, 14, 9 and 12% of the patients underwent local/regional/distant relapses or thyroid cancer-related death, respectively. Through use of the above six independent clinical variables and the parameters relating to the four clinical events and their interrelations, mean cause-specific survival probabilities of 88, 83 and 78% were determined at 10, 20 and 30 years, respectively. The survival-predicting software (PATHYPRE) written on the basis of the biostatistical model is available through Internet connections on the home page of the National Institute of Oncology, Budapest (www.oncol.hu). CONCLUSION: Prediction of the individual survival probability for extrastudy cases affords a rationale for individualization of the treatment of papillary thyroid cancer patients.

Epithelial mesothelioma with deciduoid features.

A rare case of malignant mesothelioma in a 15-year-old girl is described. The patient presented with secondary amenorrhoea and clinical symptoms resembling those of an ovarian cyst. One large and multiple small peritoneal nodules were found at laparoscopy. Histologically the tumour was characterised by an unusual pattern with a superficial resemblance to decidual reaction, but because of significant mitotic activity the diagnosis of a malignant tumour, epithelial mesothelioma with deciduoid features, was made. The patient died 11 months after diagnosis. Post-mortem examination revealed extensive extraperitoneal spread.

Effect of a single high dose and repeated small doses of dianhydrogalactitol (DAG; NSC-132313) on rat intestinal mucosa.

The influence of the treatment schedule of dianhydrogalactitol on its effect on the activity of mucosal enzymes in rat intestine was studied. The effect of a single high dose (10 mg/kg) was compared with that of repeated small doses (4 x 2.5 mg/kg) given at daily intervals. At 48 h after a single high dose the activities of thymidine kinase, which is a marker of dividing crypt cells, and of alkaline phosphatase, sucrase, maltase, xanthine oxidase, which are markers of mature enterocytes, were strongly depressed. Even 96 h after the treatment low enzyme activities could be observed. Repeated small doses caused milder enzyme inhibition and almost total recovery had occurred by 96 h after administration of the last dose. The results indicate that fractionation of drug administration can reduce the toxic side-effects on the intestinal mucosa and might be partly responsible for the higher therapeutic index of such schedules in experimental tumor models.

Secretory breast cancer in a 7.5-year old boy.

Breast cancer is extremely rare in children, and consequently no consensus has been reached on the optimal treatment modalities. The medical history and treatment plan for a 7.5-year old male breast cancer patient is described. Radical mastectomy with sentinel node biopsy was performed in October 2002. As no malignant cells were detected in the sentinel node, and no BRCA1-2 mutations were detected, no further radio- or chemotherapy was performed. A "wait-and-see" policy was decided on. Further treatment will be given if this becomes necessary with the development of metastases.

Desmoplastic small round cell tumour of the pleura: a case report with further cytogenetic and ultrastructural evidence of 'mesothelioblastemic' origin.

A typical case of desmoplastic small round cell tumour of the pleura in a 25-year-old man is described. In addition to the typical histological and immunohistological findings, the EWS-WT1 fusion product was also observed. Ultrastructurally, some tumour cells displayed intracytoplasmic neolumina, with short microvilli characteristic of submesothelial cells. These findings support the theory of a splanchnopleuric intraembryonic mesodermal, 'mesothelioblastemic' origin, both morphologically and cytogenetically.

Non-isotopic in situ hybridization of human papilloma virus on histologic sections: an amended protocol.

The authors report on their experience with an HPV non-radioactive in situ hybridization kit and describe the favorable results gained with the amended protocol, which are as follows: 1. The application of a decreased amount of both the probe and the chromogen substrate did not alter the quality of reactions. Therefore we were able to make 60 reactions instead of the originally suggested 21. 2. The proteolytic enzyme digestion time could be prolonged by changing proteinase-K for pepsin which intensifies the signal of hybridization. 3. By changing the order of hybrid detection and posthybridization washing, we succeeded in removing the excess amount of probe-ABC-AP-BAAV-ABC-AP conglomerates without losing the target sequence. 4. Using alkaline phosphatase or ABC-AP-BAAV-ABC-AP complex instead of peroxidase it was possible to demonstrate a very low number of gene copies, even if they were not detectable following the original instructions.

DNA cytometry and morphometry by TV based image analysis system (TAS) in the diagnosis of gastric carcinoma.

Cytological smears from gastric biopsies were studied by two types of TAS. Böcking's algorithm and a great number of form factors were used to separate the intact cells from the tumourous ones. The numerically expressed density features and morphometric parameters are able to indicate the possibility of malignant transformation even in dysplasia. These values become much higher in tumours and lead to unequivocal diagnosis. Both types of TAS render valuable help in assessing cases not reliably diagnosed by routine staining procedures.

Enzymological and morphological changes in rat intestinal mucosa following treatment with alkylating sugar alcohol derivatives.

Wistar rats were treated with alkylating sugar alcohol derivatives, dianhydrogalactitol (DAG) and diacetyldyanhydrogalactitol (Diac-DAG), respectively. The drugs were intravenously administered as a single, bolus injection. The applied doses 2.5, 5, 10, 17 mg/kg DAG and 5, 10, 20, 40 mg/kg Diac-DAG were roughly equitoxic. The effect of these cytostatic agents was studied on the different marker enzymes (thymidine kinase, xanthine oxidase, alkaline phosphatase, sucrase, maltase) of the separated mucosa cells derived from the functional and proliferating zone of the small intestine. Both DAG and Diac-DAG inhibited the enzyme activities of the proliferating and mature enterocytes in a dose dependent fashion, primarily acting on the crypt specific thymidine kinase. The time dependent sequence in the biochemical alterations correlated well with the cytomorphological changes. The drug-induced damage was most pronounced 48 hours after a single treatment. The regeneration of the intestinal mucosa began on days 3 and 4 and was completed by day 7. Diac-DAG at equimolar concentration proved to be more toxic than DAG on the intestine as judged by the significantly higher decrease of protein content and xanthine oxidase activity.

Prognostic significance of P53 histochemistry and DNA histogram parameters in head and neck malignancies.

BACKGROUND: The aim of the study was to determine the role of quantitative pathological parameters in prognosis of head and neck malignancies. MATERIALS AND METHODS: 51 head and neck squamous cell carcinoma patients were examined for mutant p53 gene expression (45 out of 51 patients) by immunohistochemistry and for cellular DNA-content (44 out of 51 patients) using digital picture analyzer. Statistical analysis was performed using BMDP package. RESULTS: No correlation with prognosis was found for age, sex, localization, T-classification and therapy. There was significant relationship between N-status and overall survival (p = 0.0008). No correlation was found with overall and disease-free survival for either histologic type or grading. P53: No significant correlation was detected with overall survival. A relationship was found between mutant p53 and metastasis-free time (p = 0.06). Ploidy: There were no significant differences between aneuploid and euploid tumors for either disease-free or overall survival. Synthetic (S)-phase fraction: A correlation was found for both survival rates (p = 0.029) and metastasis-free time (p = 0.05). Polyploid fraction (PF): correlation was shown for both overall survival (p = 0.0128) and metastasis-free time (p = 0.0038). CONCLUSION: There is correlation between p53 overexpression and metastatic potential and there is a significant relationship between SPF and PF value and prognosis (metastasis-free and overall survival) of head and neck cancer.

Hepatocyte proliferation and cell cycle phase fractions in chronic viral hepatitis C by image analysis method.

OBJECTIVE: Chronic hepatitis is characterized by necrosis of liver cells, accompanied by an inflammatory reaction and compensatory cell proliferation. The interaction of the core and non-structural proteins of hepatitis C virus (HCV) with several cellular factors suggests that cell proliferation may be influenced by HCV. The aim of this study was to investigate hepatocyte proliferation and DNA ploidy patterns in patients with chronic viral hepatitis C (CH-C) compared with chronic non-viral hepatitis (CH-N), using a TV image analysis method. METHODS: The DNA index (DI) and cell phase fractions (G1, S, G2) were measured by means of digital picture analysis method on nuclear suspensions of Feulgen stained hepatocytes. Cells were taken from the liver biopsy specimens of 71 patients with CH-C and 24 patients with CH-N. Twenty-six normal liver samples were used as controls. RESULTS: Significantly higher G1 (94 +/- 4) and lower S (3.56 +/- 3.16) phase fractions were measured in CH-C compared with CH-N (G1, 90 +/- 6; S, 6.4 +/- 5.99). The DI of moderate (1.12 +/- 0.05) and severe (1.12 +/- 0.05) CH-C showed near-aneuploid DNA content, while diploidy (DI < 1.10) was detected in cases of CH-N. CONCLUSION: The higher G1 and lower S cell cycle phase fractions in CH-C reflect decreased hepatocyte proliferation compared with CH-N. The near-aneuploid DNA content of the HCV-infected liver samples may be a sign of increased genetic instability, which may contribute to the carcinogenic potential of HCV.

Unusual benign neurogenic soft tissue tumour. Epithelioid schwannoma or an ossifying fibromyxoid tumour?

An unusual form of subcutaneous soft tissue tumour with benign character is reported. The tumour was composed of uniform cells which were epithelioid in appearance. True myxoid matrix, osteoid formation or metaplastic bone were not present. Immunohistochemically S-100 protein and vimentin was demonstrated in the tumour cells. Electron microscopic examinations showed fragmented or continuous basal lamina, interdigitating cytoplasmic projections, myelin-like figures and a few cell junctions. The immunohistochemical and electron microscopic results indicate that the tumour is of Schwann's cell origin. Based on light microscopic, electron microscopic and immunohistochemical results we think our case is an epithelioid benign schwannoma but the non-ossifying form of fibromyxoid tumour of the soft parts cannot be excluded with confidence.

Cytophotometric determination of basic proteins with heparin-alcian blue method.

For the quantitative cytochemical determination of basic proteins, Heparin-Alcian Blue (HAB-was found to be a more suitable staining procedure than the generally used alcalic Fast green reaction. HAB is able to stain all the histone fractions, but there is no staining following deamination (gelelectrophoretic examinations) and no aspecific staining (Millipore filter model). In EHRLICH ascites tumour cells, changes in the FEULGEN-DNA content is followed by the proportional changes of HAB-basic protein content. After HAB reaction, the FEULGEN reaction can be performed on the same slide.

Significance of histologic grading in the prognosis of ovarian tumors.

The prognostic significance of a histologic grading system was studied by an analysis of pathological specimens from 73 patients with ovarian epithelial cancers and tumors of borderline malignancies (tumors of low malignant potential) collected over a period of 25 years. The survival probability of patients in each group was analyzed by a computer program based on the product limit of Kaplan and Meier. The histologic grading scheme based on the presence of papillary formations or glands versus the presence of solid tumor nests proved to be prognostically significant for patients with stage I and II disease. The other grading system, based on cytologic features, was able to stratify survival rates into three instead of two groups distinguished by the histologic grading system. The survival of patients with moderately and poorly differentiated tumors was essentially the same. No difference was found between the prognoses of patients with cytologic grades 3 and 4.

Trisomy 12 in juvenile granulosa cell tumor of the ovary during pregnancy. A report of two cases.

BACKGROUND: Granulosa cell tumors constitute only 5% of ovarian neoplasms, and their coexistence with pregnancy is extremely rare. Juvenile granulosa cell tumor has a good prognosis if it is confined to the ovary, but this type behaves more aggressively than the adult type at advanced stages. CASES: We report on successful completion of two singleton pregnancies and deliveries of normal infants in two young women with juvenile granulosa cell tumor diagnosed and treated during pregnancy. This tumor has rarely been described in association with pregnancy. The presence of trisomy 12 as a single chromosomal abnormality was detected in these two tumors. Both tumors were localized strictly to the ovary, so conservative surgery was applied and proved sufficient to remove all tumor tissue. Follow-up showed no signs of recurrence 18 and 53 months after the interventions. CONCLUSION: These cases support the contention that trisomy 12 is a nonrandom chromosome abnormality in juvenile granulosa cell tumors and that pregnancy may affect nuclear stability in this tumor.