Prognostic value of KIT/PDGFRA mutations in gastrointestinal stromal tumours (GIST): Polish Clinical GIST Registry experience.

BACKGROUND: Majority of gastrointestinal stromal tumours (GISTs) are characterised by KIT-immunopositivity and the presence of KIT/platelet-derived growth factor receptor alpha (PDGFRA) activating mutations. PATIENTS AND METHODS: Spectrum and frequency of KIT and PDGFRA mutations were investigated in 427 GISTs. Univariate and multivariate analysis of relapse-free survival (RFS) was conducted in relation to tumours' clinicopathologic features and genotype. RESULTS: Mutations were found in 351 (82.2%) cases, including 296 (69.3%) KIT and 55 (12.9%) PDGFRA isoforms. Univariate analysis revealed higher 5-year RFS rate in women (37.9%; P = 0.028) and in patients with gastric tumours (46.3%; P < 0.001). In addition a better 5-year RFS correlated with smaller tumour size ≤ 5 cm (62.7%; P < 0.001), tumours with mitotic index ≤ 5/50 high-power fields (60%; P < 0.001), and characterised by (very) low/moderate risk (70.2%; P = 0.006). Patients with GISTs bearing deletions encompassing KIT codons 557/558 had worse 5-year RFS rate (23.8%) than those with any other KIT exon 11 mutations (41.8%; P < 0.001) or deletions not involving codons 557/558 (33.3%; P = 0.007). Better 5-year RFS characterised patients with KIT exon 11 point mutations (50.7%) or duplications (40%). By multivariate analysis, tumours with PDGFRA mutations and KIT exon 11 point mutations/other than 557/558 deletions had lower risk of progression than with KIT exon 11 557/558 deletions (both Ps = 0.001). CONCLUSIONS: KIT/PDGFRA mutational status has prognostic significance for patients' outcome and may help in management of patients with GISTs.

p21/WAF1/Cip1 expression in invasive ductal breast carcinoma: relationship to p53, proliferation rate, and survival at 5 years.

The p21/WAF1/Cipl antibody, DCS-60, was characterized by means of immunoblotting and immunofluorescence on a variety of human breast cancer cell lines. Heterogeneous staining of nuclei was observed with strong staining of cells in early G1. p21/WAF1/Cipl expression in invasive ductal, not otherwise specified breast carcinomas was determined using immunohistochemistry with this antibody and computerized image analysis. Two hundred and twenty-two tumors, including 130 from patients with no axillary node involvement, were examined. p21-positive tumor cell nuclei were found in 30% of the breast carcinomas. The percentage of tumor cell nuclei that were positive ranged from less than 1% to greater than 10%. In the whole cohort of patients, p21 expression was significantly associated with a low histological grade. In the node-negative group, there was a significant negative correlation between p21 positivity and a high (>10%) MIB-1 score. The mean MIB-1 score was significantly lower in p21-positive tumors in the whole cohort of patients (P=0.03) and in the nodenegative group (P=0.02). No association was found between p21 expression and overall survival at 5 years. With respect to p21/p53 phenotype, the significant difference in survival was noted only for the group of patients treated with adjuvant chemotherapy. The p21- p53+ phenotype had the worst survival (58% surviving 5 years), while the p21+ p53- phenotype had good survival (83% surviving 5 years; P<0.05). The results seem to suggest a correlation between p21/p53 phenotype and response to adjuvant chemotherapy.

Concerning cellular defense in carcinoma of the larynx.

In the search of microscopic manifestations of cellular defense in tumor disease, larynx specimens removed surgically for carcinoma of the larynx and neighboring lymph nodes were studied morphologically. Blastic transformation of lymphocytes from the lymph nodes was also investigated. Activity of nuclear chromatin of these cells was assessed on the basis of its sensitivity to acid hydrolysis in the reaction of Feulgen and incorporation of labeled 3H-actinomycin D. The results indicate existence of cellular defense reaction in carcinoma of the larynx, around the primary tumor as well as in neighboring lymph nodes. Intensity of the reaction in lymph nodes increases with growth of the primary tumor.

Keratin-positive reticulum cells in fine needle aspirates and touch imprints of hyperplastic lymph nodes. A possible pitfall in the immunocytochemical diagnosis of metastatic carcinoma.

Fine needle aspirates and touch imprints of 36 hyperplastic (reactive) lymph nodes were tested for the presence of keratin and desmin. Keratin-positive cells with morphologic characteristics corresponding to extrafollicular (fibroblastic) reticulum cells were found in 18% of the fine needle aspirates and 42% of the touch imprints. The number of keratin-positive reticulum cells varied from 1 to greater than 30 per slide. Desmin-positive cells with similar morphology were found in 23% of fine needle aspirates and 37% of touch imprints, and the number of such cells per slide ranged from 2 to greater than 70. The relatively frequent occurrence of keratin-positive reticulum cells in these preparations from hyperplastic lymph nodes should be taken into account if keratin antibodies are used to search for carcinoma micrometastases.

New educational methods in cytopathology: a transnational training program in cervical cytology--CYTOTRAIN CD-ROM.

Among the reasons for the low effectiveness of preventive measures for early detection of cancers and preinvasive states of the uterine cervix there is insufficient training of people performing cytological screening. In order to decrease the rate of false negative diagnosis it is necessary to improve the organisation of permanent professional training of cytopathologists and cytotechnicians.

Immunocytochemical evaluation of gastric mucosal cathepsin D in peptic ulcer.

The presence of cathepsin D was assessed in epithelial and stromal cells of the gastric mucosa in patients with ulcer disease depending on the course of ulcer healing, accompanying Helicobacter pylori infection, mode of treatment, location of ulceration, the presence of relapses, intestinal metaplasia, ulcer diameter, age and gender of the patients. The presence of cathepsin D was assessed by immunohistochemistry using an immunoenzymatic method StreptABC. Immunohistochemistry was performed on formalin-fixed paraffin sections obtained from 54 selected patients (20 men and 34 women, aged from 23 to 75 years), in whom endoscopy of the upper alimentary tract revealed ulcerations within the stomach, and then ulcer healing was monitored by endoscopy and histopathology. Before immunohistochemistry each specimen was routinely stained with hematoxylin and eosin, and histopathological study was performed to confirm a nonneoplastic character of ulcers, followed by staining with Giemsa method for the presence of Helicobacter pylori. The presence of cathepsin D was analyzed using image analysis. Student's t-test and a correlation coefficient were used to study the relationship between the expression of cathepsin D and selected parameters. The following significant relationships were observed: 1. between the expression of cathepsin D in epithelial cells and the course of peptic ulcer healing after antiulcerative treatment; 2. between the expression of cathepsin D in stromal cells after antiulcerative treatment and the course of ulcer healing.

Proliferation cell nuclear antigen (PCNA) expression in cervical intraepithelial neoplasia (CIN). An immunohistochemical study.

We analyzed the expression of proliferating cell nuclear antigen (PCNA) in ectocervical sections fixed in formalin and embedded in paraffin using monoclonal antibody PC10, PAP and APAAP methods. A total of 156 cases was studied: 32 cases with normal epithelium, 33 cases with CIN I, 36 cases with CIN II, 55 cases with CIN III. We evaluated: 1. the index of PCNA (IPCNA), i.e. the proportion of PCNA-positive cells relative to all cells in the epithelium; 2. the IPCNA in each epithelial layer; 3. the epithelial height with PCNA-positive cells relative to the total height of the epithelium; 4. changes in the IPCNA with respect to the presence or absence of histological signs of HPV infection. We found: 1. a significant correlation between the index of PCNA and the severity of dysplasia in the whole epithelium and its individual layers; 2. a significant correlation between the epithelial height with PCNA-positive cells relative to the total height of the epithelium and the severity of dysplasia; 3. a lack of correlation between the IPCNA and HPV(+) and HPV(-) cases.

Nuclear morphometry of MIB-1 positive and negative tumor cells in primary and metastatic malignant melanoma of the skin.

The purpose of this study was the evaluation of nuclear area, nuclear axis ratio, perimeter and roundness of nuclei of tumor cells with and without Ki-67 antigen expression (demonstrated immunohistochemically using MIB-1 antibody) in primary and metastatic malignant melanoma of the skin. The parameters were further analyzed with respect to their association with the depth of malignant invasion according to Clark [7] and tumor thickness according to Breslow [6]. 142 malignant melanomas (53 primary and 89 metastatic), were assessed employing a computerized image analyzer Quantimet 600S (Leica). The mean nuclear area of MIB-1 positive nuclei was significantly larger than that of the negative ones (p < 0.0001) both in primary and metastatic malignant melanoma. In comparison to the primary melanoma the nuclei of metastatic melanoma cells had a larger area and were more rounded, while the MIB-1 positive nuclei additionally showed a greater degree of polymorphism of their area and shape. With growing invasion thickness according to Breslow and increased Clark's level, the mean nuclear area of tumor cells increased, and their shape became more round. The MIB-1 positive tumor cell nuclei of primary melanomas with metastases were significantly out of round in comparison to primary melanomas without metastases. The results indicate an association between the area and shape of melanoma cell nuclei and the presence of metastases, and between the nuclear area of tumor cells and such factors related to poor prognosis as the depth of invasion and the tumor thickness.

Quality control in cytodiagnosis of carcinoma of the cervix.

In many European countries the morbidity and mortality due to cancer of the cervix are too high. Even in countries, where smears have been taken at regular intervals for many years, the percentage of new cases does not decrease as much as it was expected. There are several reasons that may explain this situation. At least two of them can be solved by pathologists: quality control (QC) system and organization of a school of gynaecological cytopathology for pathologists and cytotechnicians. Here we focus on the main aspects of External and Internal QC and new approaches to QC, including new methods and stricter control of the professional level of all persons involved in the screening and diagnosis of cervical cancer.

PCNA defined proliferative activity of epithelial tumor cells in adenomas of the colon.

Proliferative activity of epithelial tumor cells was evaluated with the use of immunohistochemistry and anti-PCNA monoclonal antibodies in alcohol fixed paraffin embedded sections of 44 colonic adenomas, including 33 tubular, 5 villous and 6 tubulovillous adenomas. The mean PCNA index was 24.7 +/- 10.9%, 24.8 +/- 6.2% and 24.8 +/- 14.0% in tubular, villous and tubulovillous adenomas respectively. In 12 tubular adenomas with dysplasia the mean PCNA index in areas with dysplasia was significantly higher (38.2 +/- 11.5%) as compared to areas without dysplasia (17.0 +/- 8.9%; p < 0.05). The results indicate that PCNA index of epithelial tumor cells is significantly increased in adenomas with high grade of dysplasia irrespective of histological type or size of the tumour.

Tumour cell proliferation rate as determined by MIB-1 antibody in Wilms' tumour.

Determination of neoplastic cell proliferation becomes an important and objective element of assessing malignancy of neoplasms and their response to therapy. The basic problem in the management of Wilms' tumours in children is selection of patients to appropriate risk groups. The purpose of the present study was to evaluate proliferation index in 35 Wilms' tumours determined immunohistochemically by using monoclonal antibody MIB-1. The final analysis included 39 preparations of formalin fixed and paraffin embedded tissue sections. Proliferation index of neoplastic cells was determined in blastema, epithelium and stroma of the tumours. We found a marked difference between low proliferation index in stromal cells and high proliferation index in blastemal and epithelial cells. In tumours after chemotherapy we found higher proliferation index in epithelial cells and lower index in blastemal cells as compared to tumours before chemotherapy. Higher proliferation index in blastemal cells and epithelial cells was associated with worse prognosis. Worse prognosis was seen in cases in which after chemotherapy proliferation index in blastemal cells was still high.

Fhit protein expression in endometrial cancers: no correlation with histological grade.

The genes specifically involved in endometrial cancers have not yet been discovered. The FHIT gene, a tumour suppressor located at 3p14.2, is altered in many human tumours, including those derived from the female genital tract. We have thus investigated the status of Fhit protein expression in endometrial carcinomas (EC), and its association with histological grade of malignancy in order to determine if Fhit expression is inactivated in EC and if so, whether it is inactivated during initiation or progression. Recent studies have reported that alteration in the FHIT locus detected by DNA and RNA analysis is well correlated with loss of Fhit protein expression in tumours. Thus, we characterised Fhit protein expression as an indication of FHIT gene status in 35 cases of EC of different histological grade (G1: 13 cases; G2: 14 cases; G3: 8 cases). In our group of cancers, Fhit protein expression was absent or reduced in 37% (13/35) of EC. The first 13 cases, judged as G1, showed Fhit deficiency in approximately 38.5% of cases (5/13). For G2 and G3 tumours these numbers were similar and accounted for approximately 35.7% (5/14) and approximately 37.5% (3/8), respectively. No statistical difference was found for Fhit expression among the various groups of tumours, which allowed us to conclude that morphological grade does not seem to be an important factor. Our results suggest that Fhit inactivation is an early event in carcinogenesis of the endometrium. As this observation is contrary to some already published reports, another independent study with larger amounts of material is necessary to determine this issue definitely.

[Contribution of immunocytochemical marking of benign cells to the diagnosis of serous effusions]. / Apport du marquage immunocytochimique des cellules bénignes au diagnostic des épanchements des séreuses.

In pleural and peritoneal effusions, the distinction between mesothelial cells and histiocytic cells is fairly often difficult. This distinction is specially important in malignant effusions, where such reactive cells can be confused with tumoral ones. In order to increase the accuracy of cytological diagnosis, the immunocytochemical reactivity with 13 antibodies was studied. Four antibodies (vimentin, lysozyme, monoclonal keratin, OKM1) realise a panel specially useful for the characterization of mesothelial and histiocytic cells in most cases of benign or malignant effusions.

[Argon plasma coagulation in a patient with early diagnosis of gastric carcinoma]. / Argonowa koagulacja plazmowa u chorego z wczesnie rozpoznanym rakiem wpustu.

Argon plasma coagulation (APC) has been introduced for the local endoscopic treatment of gastrointestinal malignancy recently. It is mainly used as a palliative therapy, especially in case of stenosis. Despite a lot of publications concerning APC the clinical usefulness of this method in a small malignant tumors remains unclear. The patient with early diagnosed carcinoma of gastric, efficiently treated using argon plasma coagulation is described.

[A patient with abdominal actinomycosis--diagnostic problems]. / Chora z promienica brzuszna--problemy diagnostyczne.

The difficulties in diagnosis of abdominal actinomycosis are presented. Clinical manifestations and colonoscopy suggested malignancy. Final diagnosis was made on the basis of pathological assessment of resected sigmoid. Authors underline that in case of "negative" pathomorphological results of material obtained during endoscopy from lesions suspected, benign disease should be consider including anctinomycosis and intraoperative pathomorphological examination should be performed.

[Usefulness of bronchoalveolar gavage in patients with acquired immunodeficiency syndrome]. / Ocena przydatnosci badania popluczyn oskrzelowo-pecherzykowych u chorych z zespolem nabytego braku odpornosci (AIDS).

51 BALFs from HIV positive patients were analysed. In 16 of them Pneumocystis carinii organisms were found. Two methods detecting Pneumocystis carinii were used: Grocott-Gomori, fluorescence Papanicolaou stain. The best staining method turned out to be Grocott-Gomori. The differential count was carried out in 40 BALFs. In patients with Pneumocystis carinii detected by BAL neutrophilia was always present. In some of these patients lymphocytosis was also present. In 4 patients with high lymphocytosis immunocytochemical examinations with monoclonal antibodies against CD4, CD8 T cells showed a marked decrease of the CD4/CD8 ratio.

[Diagnostic value of bronchoalveolar lavage fluid examination in various interstitial lung diseases]. / Wartosc diagnostyczna badania popluczyn oskrzelowo-pecherzykowych w róznych chorobach sródmiazszowych pluc.

The results of 229 routine BAL examinations in patients with different interstitial lung diseases are presented. The authors analyse the diagnostic usefulness of BAL in the differential diagnosis in interstitial lung diseases. It seems that BAL is helpful in making a specific diagnosis of interstitial lung disease.

[Subpopulations of T lymphocytes in bronchoalveolar lavage (BAL) fluid of different interstitial lung diseases]. / Subpopulacje limfocytów T w popluczynach oskrzelowo-pecherzykowych (bal) w róznych chorobach srodmiazszowych.

Immunocytochemical examinations using the PAP method with monoclonal antibodies against lymphocytes T (CD3, CD4, CD8) were carried out in 31 BAL's from patients with different interstitial lung diseases. A distinct increase of the CD4 (T "helper"): CD8 (T "suppressor") ratio was found in patients with sarcoidosis and in a patient with pigeon breeder's disease. A profound decrease of the CD4/CD8 ratio was found in HIV positive patients. The authors also discussed the methodological problems of correct typing of T cells in BAL.

[Immunocytochemistry in the differential diagnosis of the head and neck tumors]. / Badania immunocytochemiczne w diagnostyce róznicowej nowotworów glowy i szyi.

Immunocytochemical analysis in fine-needle aspirates were performer in 46 cases of head and neck tumors. Immunocytochemical studies helped to establish the differential diagnosis of all diagnostically difficult cases. The application of 3 monoclonal antibodies against keratin, vimentin and LCA permitted to differentiate basic types of malignant tumors.