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In primary glomerulonephritis the degree of interstitial fibrosis governs the renal function. Mast cells participate in renal interstitial fibrosis, but its role remains poorly understood. Some of human mast cells contain chymase and chymase may participate in local angiotensin formation. Material consisted of 35 renal biopsies. The diagnoses included diabetic nephropathy, mesangial glomerulopathy, IgA glomerulopathy and membranous glomerulopathy. Chymase and tryptase-positive cells were stained by immunohistochemistry and counted. Relative interstitial volume (RIV) was measured by point counting method. The density of tryptase-positive cells was 5.26 per 10 high power fields; the density of chymase-positive cells was 2.72. The counts were higher than in controls and highest in diabetic nephropathy. Creatinine serum level was related to density of chymase-positive cells (R = 0.57), density of tryptase-positive cells (R = 0.59) and RIV (R = 0.77). On multiple regression analysis creatinine level was influenced by RIV but also by density of chymase-positive cells. Our findings indicate that both types of mast cells are present in renal interstitium in diabetes and glomerulonephritis, and may influence the renal function. Chymase-positive cells may be more important in this regard.

We investigated the prevalence of the JAK2 V617F gain-of-function mutation in patients with Philadelphia chromosome-negative chronic myeloproliferative disorders (Ph- MPD) and explored the links between JAK2 mutational status and the clinicopathologic picture of essential thrombocythemia (ET), chronic idiopathic myelofibrosis (CIMF), and polycythemia vera (PV). Allele-specific polymerase chain reaction results for 59 ET, 18 CIMF, and 9 PV cases were compared with values for clinical variables at presentation and last follow-up and with the diagnostic trephine bone marrow biopsy pictures. JAK2 V617F was found in 38 (64%) of ET cases, 7 (39%) of CIMF cases, and 9 (100%) of PV cases. The ET patients with the mutant JAK2 showed significantly higher (although not overtly polycythemic) red blood cell parameter values, lower platelet counts, and higher white blood cell counts. Similar trends were found in CIMF. Megakaryocyte clustering was much less pronounced in the CIMF cases with mutant JAK2, with an analogous trend occurring in the ET cases. Bone marrow cellularity values and the numbers of CD34+ and CD117+ blasts in the ET and CIMF groups did not differ. Fibrosis was slightly less marked in the ET cases with mutant JAK2. The mutation did not significantly influence the clinical course during the follow-up in either disease in the short term (median follow-up, 22 months). The JAK2 V617F mutation is prevalent in all Ph- MPD and may skew their presenting phenotype, including bone marrow histology, toward a more “erythremic” and less “thrombocythemic” phenotype.

Buccal epithelial cells are occasionally used as a source of supposedly non-neoplastic DNA in patients suffering from haematological malignancies. Formerly, Kralovics et al found the JAK2 V617F mutation in DNA derived from buccal swabs in only 2.2% of patients with Philadelphia-negative chronic myeloproliferative disorders (Ph− CMPD).1 We compared the JAK2 V617F mutational status in DNA derived from buccal swabs to that in DNA extracted from either bone marrow or peripheral blood in 35 Ph− CMPD patients, including five cases of polycythaemia vera (PV), five cases of chronic idiopathic myelofibrosis (CIMF) and 25 cases of essential thrombocythaemia (ET). Genomic DNA was isolated from buccal swabs immediately on collection and …

The term glomerulonephritis encompass a heterogeneous group of diseases; these are a important cause of end stage renal disease. Although several evidence exist, that the main prognostic factors are extraglomerular lesions, no quantitative assessment is usually done. In nephropathological practice a semiquantitative approach is preferred. However, most of work on extraglomerular lesions significance was done with quantitative methods. The aim of the study was to compare the effects of quantitative and semiquantitative assessment of extraglomerular lesions in glomerulonephritis. The material consisted of 120 renal biopsies. On inspection, percentage of sclerosed glomeruli, degree of interstitial fibrosis, degree of interstitial infiltration, degree of tubular atrophy were and degree of mesangial matrix expansion assessed. For quantitative measurements AnalySIS 3.0 pro image analysis system was used. Relative interstitial volume, volume of interstitial infiltrate, with their variability--ross sectional areas of proximal and distal tubules were assessed by point counting method. Relative interstitial volume was significantly correlated to percentage of sclerosed glomeruli (R = 0.33 p0.001), degree of tubular atrophy (gamma = 0.57 p0.00001), degree interstitial fibrosis (gamma = 0.31 p0.0002) and mesangial matrix expansion (gamma = 0.24 p0.001). Semiquantitative and quantitative assessment of interstitial infiltrate was significantly correlated as well (gamma = 0.81 p0.001). Semiquantitatively assessed degree of tubular atrophy showed significant relation to total proximal tubular area (gamma = -0.30 p = 0,004). Percentage of sclerosed glomeruli was significantly correlated to creatinine level (R = 0.24 p = 0.03), but not to urea level (R = 0.09, NS). Semiquantitatively assessed degree of interstitial fibrosis showed only marginal correlation to creatinine level (gamma = 0.18 NS), however degree of interstitial infiltration was significantly correlated to creatinine (gamma = 0.34 p = 0.002) and urea level (gamma = 0.22 p = 0.06). Degree of tubular atrophy was significantly correlated to creatinine (gamma = 0.43 p0.001) and urea level (gamma = 0.28 p = 0.015). Relative interstitial volume was the very most important correlate of creatinine (R = 0.47 p0.0001) and urea level (R = 0.30 p0.01). In conclusion, it was confirmed, that the strongest correlate of renal function is relative interstitial volume. Some, but not all of semiquantitative parameters are also significantly correlated to kidney function.

Glomerulonephritis is one of the diseases leading to chronic renal failure and need of renal replacement therapy. Changes in extraglomerular compartments, especially in the interstitium, are thought to play a major role in progression. However, the exact relationships between renal interstitium, tubules and vessels and their prognostic impact are less well understood. The material consisted of 111 biopsies with primary glomerulonephritis. Normal renal tissue from surgically removed kidneys served as controls. Relative interstitial volume (RIV), its variability, volume of interstitial infiltrate, cross-sectional tubular area were measured with the point-counting method. A number of vascular parameters were also measured. The assessed interstitial and tubular parameters were strongly correlated to creatinine level. The strongest correlation was seen for RIV, also on multiple regression. In patients with renal failure, increased RIV, more pronounced vascular lesions and interstitial infiltrates were seen. Survival analysis showed that interstitial expansion is the most important factor leading to renal failure. Tubulointerstitial and vascular factors are interrelated and linked to renal function. RIV has strongest impact on renal function and survival, even taking into account other factors.

Immunophenotype of isolated (disseminated or circulating) tumour cells (ITC) in the blood, bone marrow and lymph nodes were studied in patients with gastric cancer. Coexpression of metalloproteinases inducer (EMMPRIN), chemokine receptors (CCR6, CXCR4) and adhesion molecules (Ep-CAM, CD44) was determined on cytokeratin positive (CK+) cells in CD45- cell population sorted out from the blood and/or bone marrow. Eight cytospin samples of blood and 69 samples of bone marrow containing CK+ cells from patients with gastric cancer were included into study. Expression of EMMPRIN and CCR6 were noted in a half of CK+ samples (of blood/bone marrow) whereas the expression of CXCR4 and Ep-CAM was much lower. Analysis of paired data of these determinants expression on CK+ cells showed no association between them. Expression of EMMPRIN, Ep-CAM, CCR6, CCR7, CXCR1, and CXCR4 on ITC in lymph nodes was determined by flow cytometry. In 18 lymph nodes (out of 36 assayed) CK+ cells were found. The expression of CCR6 and Ep-CAM on CK+ cells was observed in almost all studied lymph nodes, CXCR1--in half of them. The expression of EMMPRIN and CCR7 cells was lower. These results suggest that ITC of gastric cancer express variably several molecules that may be involved in metastasis formation.

Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of gastrointestinal tract. GISTs range from benign indolent neoplasms to highly malignant sarcomas. Gain-of-function mutations of tyrosine kinase receptors, KIT or PDGFRA, have been identified in most GISTs. In this study, we report 36 GIST patients whose tumors had homozygous KIT exon 11 mutations detected by direct sequencing of PCR products. Loss of heterozygosity in KIT locus and other chromosome 4 loci were documented in majority of these tumors. However, fluorescence in situ hybridization with KIT locus-specific probe and chromosome 4 centromeric enumeration probe showed no evidence of KIT hemizygosity in a majority of analyzed cases. These findings are consistent with duplication of chromosome 4 with KIT mutant allele. Homozygous KIT exon 11 mutations were found in 33 primary tumors and 7 metastatic lesions. In two cases, shift from heterozygosity to homozygosity was documented during tumor progression being present in metastases, but not in primary tumors. Among primary GISTs, there were 16 gastric, 18 intestinal and 2 from unknown locations. An average primary tumor size was 12 cm and average mitotic activity 32/50 HPFs. Out of 32 tumors 29 (90.6%) with complete clinicopathologic data were diagnosed as sarcomas with more than 50% risk of metastatic disease, and 26 of 29 patients with follow-up had metastases or died of disease. An average survival time among pre-imatinib patients, who died of the disease was 33.4 months. Based on these findings, we conclude that presence of homozygous KIT exon 11 mutations is associated with malignant course of disease and should be considered an adverse prognostic marker in GISTs.

One of the prognostic and predictive factors in invasive breast carcinomas is determination of the HER2/neu proto-oncogene amplification or HER2 protein overexpression. HER2 amplification/overexpression is associated with a more aggressive disease course, greater likelihood of recurrence and generally poor prognosis. The authors compared the specificity, simplicity of a given procedure and method standardization, the simplicity of evaluation the results of each in situ hybridization method and time needed for performing the test. Sixty-three cases of infiltrating breast carcinoma from surgically excised tumors and core needle biopsies were included in the study. The first step was the determination of HER2 status by immunohistochemistry. The patients with moderate (2+) and strong (3+) overexpression of HER2 protein were chosen for determining HER2 amplification by three methods of in situ hybridization: FISH, CISH and in situ hybridization with silver autometallography. The statistical analysis revealed a good agreement between IHC and ISH methods and among ISH methods. The results indicate that all in situ hybridization methods are equivalent tools for evaluating HER2 gene amplification in archival material. There is no clear answer which method is the best assay to determine HER2 marker status, although the authors present some advantages and disadvantages of all the described techniques and a proposed algorithm for choosing a method for a given laboratory.

Recent reports indicate that the alterations in the p16 and pRb pathways can influence tumour progression and poor prognosis in several tumours. The objective of this study was to analyse p16 and pRb expression in161 patients with malignant fibrous histiocytomas (MFH). By immunohistochemistry, p16 and pRb were demonstrated in 25% and 56% of MFH, respectively. Cox regression analysis demonstrated an independent prognostic influence of both genes. Generally, the loss of p16 and pRb expression correlated with poorer prognosis. Promoter methylation of p16 was found in 16/42 of p16 negative MFH and of pRb in 2/42 of pRb-negative MFH. It can be concluded that p16 and pRb alterations play an important role in the progression of soft tissue sarcomas.

Colorectal carcinoma is etiopathologically heterogenic. It may develop through a sequence of mutations leading to chromosome instability or be a result of defects in DNA repair mechanisms manifested by microsatellite instability of varying degrees. Colorectal carcinoma can thus be classified into microsatellite-stable (MSS), highly microsatellite unstable (MSI-H) and intermediate low-level microsatellite unstable (MSI-L) groups. Fluorescent hybridization in situ (FISH) is a method of detecting specific sequences of nucleic acids that is based on specific bonding of a fluorescent marker-associated probe and specific DNA fragment. The material consisted of 146 non-selected cases of colorectal carcinoma patients operated on at First Chair of General Surgery, Collegium Medicum, Jagiellonian University in Cracow, Poland. Following a standard histopathological evaluation, tissue microarrays were prepared using a Tissue MicroArray Builder, and FISH was performed employing probes specific for chromosomes 1, 8, 17 and 18. Microsatellite instability was evaluated in frozen material using the PCR reaction with gel and capillary electrophoresis. The mean number of signals obtained for chromosome 1 in the entire material was 2.06, while the corresponding mean values in the MSS group equaled 2.07, in the MSI-L group - 2.07, and in the MSI-H group - 2.01. The mean number of signals for chromosome 17 in the entire material was 2.1, in the MSS group - 2.11, in the MSI-L group - 2.13, and in the MSI-H group - 2.01. The number of signals for chromosome 18 in the entire material was 2, in the MSS group - 2, in the MSI-L group - 2, and in the MSI-H group - 2. The means number of signals for chromosome 8 in the entire material was 2.07, in the MSS group - 2.08, in the MSI-L group - 2.01, and in the MSI-H group - 2. These differences are not sufficient for distinguishing colorectal carcinoma molecular forms.

Colorectal carcinoma is a frequent malignant tumor, characterized by varying clinical course and response to treatment. At the molecular level, colorectal carcinomas are divided into tumors with chromosomal instability (microsatellite-stable, MSS), microsatellite instability (MSI-H) and low microsatellite instability (MSI-L). The method of tissue microarrays allows for combining materials originating from multiple patients into a single slide, what makes possible to simultaneously investigate large material for the presence of numerous, diversified markers. The study material consisted of 208 cases of colorectal carcinoma. Microsatellite instability was evaluated in frozen material employing the PCR reaction with gel and capillary electrophoresis. Following a standard histopathological assessment, tissue microarrays were prepared using a MTA-1 microarrayer (Beecher) and standard immunohistochemical reactions were performed to detect the presence of bcl-2, CDX-2, Ki67, MLH1, MSH2, MSH6, p16, p53 and survivin. Apoptotic cells were detected using the TUNEL method. The correlations between the reactions were investigated and differences in the expression of the investigated proteins noted in carcinomas with various degrees of microsatellite instability. The agglomeration analysis showed differences in patterns of expression between MSS, MSI-L and MSI-H carcinomas. The discriminant function analysis demonstrated that the MSI-H carcinomas were best differentiated by MLH1, survivin and Ki67 expression, while the MSI-L tumors differed from the remaining colorectal carcinomas by their apoptotic index, local tumor stage (pT), the presence of angioinvasion and mucin production.

Alterations of DNA mismatch repair genes, primarily demonstrated in hereditary nonpolyposis colorectal carcinomas, were reported to be of relevance for the progression of several sporadic tumours. In this study, the expression and mutations of MLH1, MSH2, PMS1 and PMS2 in a panel of thyroid tumours, including nodular hyperplasia, follicular adenomas and carcinomas, were investigated. The expressions of MLH1, MSH2 and PMS1 were generally higher in malignant tumours than in benign lesions (p0.01). This observation can find potential diagnostic application in the differentiation of follicular adenomas from follicullar carcinomas of the thyroid. No point mutations in the DNA mismatch repair genes MSH2 (exon 12, 13) and MLH1 (exon 15, 16) were found.

The development of new blood vessels is a prerequisite for progression of malignant neoplasms. Factors that induce neoangiogenesis include VEGF, VEGF-C, VEGF-D, PD-ECG, ANG-2, TSP-1, HIF-1 and HIF-2. From the etiopathogenetic viewpoint, colorectal carcinoma is heterogenic. It may develop via a sequence of mutations leading to chromosome instability or else result from DNA repair defects, which are manifested as microsatellite instability. The objective of the present investigations was the comparison of neoangiogenesis in microsatellite-stable colorectal carcinomas, as well as in tumors with low and high instability levels. The material included 71 surgical cases of colorectal carcinoma. Vessel density was assessed by immunohistochemical reactions to CD34 and vWf, calculating the number of vessel sections within the invasion margin, in visual fields selected at random, and within hot spots. Microsatellite instability was evaluated in frozen materials employing the PCR reaction with gel and capillary electrophoresis. In all the cases, the authors detected CD34+ and less numerous vWf+ vessels within the tumor and in its vicinity. In 45 cases, no microsatellite instability was found, in 13 cases, low level instability (MSI-L) was observed, and in another 13 - high microsatellite instability (MSI-H). Some differences in vessel density were noted between the above groups, yet they were not statistically significant. On the other hand, the authors observed more numerous CD34+ vessels in cases with metastases to the regional lymph nodes. In conclusion, it is suggested that neoangiogenesis in sporadic colorectal carcinoma is directly related to metastatic potential, but not to MSI status.

The authors present the true incidence of non-Hodgkin's lymphomas basing on the cases diagnosed in Małopolska district during one-year period. The data point to higher lymphoma morbidity than the incidence of them reported by National Cancer Register. Noteworthy is also the distribution of lymphoma types in Polish population being different from that one reported in majority of other Europe countries. The present report indicate the lymphomas become growing diagnostic and clinical challenge, as they place among the most frequent neoplasms in population.

Colorectal carcinoma is etiopathologically heterogenic. It may develop through a sequence of mutations leading to chromosome instability or be a result of defects in DNA repair mechanisms manifested by microsatellite instability. Carcinomas of this type are supposed to be characterized by a better prognosis and a different response to chemotherapy. The main target of 5-fluorouracil (5-FU) treatment is thymidylate synthase (TS). High TS expression has been identified as promoting resistance to 5-FU. The objective of the present investigation was to determine whether microsatellite instability is associated with thymidylate synthase expression. Ninety-eight cases of colorectal carcinoma were studied. Microsatellite instability was evaluated in frozen material employing the PCR reaction with gel and capillary electrophoresis. TS expression levels were assessed in preparations stained immunohistochemically using a semiquantitative method on a scale with scores from 0 to 3. The MSI-H phenotype was detected in ten cases, MSI-L in 16, and MSS in 72. The mean TS expression score was 1.79. In the MSS group, the mean TS expression score was 1.69, in the MSI-L group the mean TS expression score was 1.73, and in the MSI-H group the mean TS expression score was 2.67. The differences between MSI-H and MSS/MSI-L were statistically significant (p0.0002 and p0.004, respectively). The results may explain the different response of MSI-H carcinomas to 5-FU treatment.

The mutations of MLH1 and MSH2 have been reported to be responsible for malignant transformation and tumour progression in several sporadic tumours. Eighty-six primary malignant melanomas with known follow-up were investigated. Point mutations of DNA mismatch repair MLH1 and MSH2 in malignant melanomas were not found. Exon 12 (MSH2) was not present in 26 out of the 86 melanomas and exon 13 (MSH2) was lost in 25 of the tumours. The loss of exon 15 (MLH1) was observed in 22 out of the 86 tumours and the loss of exon 16 (MLH1) in 24 melanomas. The loss of exons correlated strongly with the loss of MLH1 and MSH2 protein expression. In multivariate analysis, including all 4 exons and expressions of MLH1 and MSH2, prognostic significance was found only for loss of exon 12 (MSH2) and loss of exon 15 (MLH1).

Reports of 1938 trephines submitted from five institutions over a 30-month period were analyzed looking for associations between the hospital of origin, operator, bone marrow pathology, patient's age and the biopsy quality. The arbitrary adequacy criteria (min 10 mm of interpretable marrow or min 10 intertrabecular spaces) were fulfilled by 61.9% of the biopsies. The performance of individual operators varied from 15.9% to 87.8% of adequate trephines. The group of doctors performing more than 100 biopsies in the study period had satisfactory results. The intermediate group (20-100 biopsies) was the least homogenous, and on the average had the poorest biopsy quality. The biopsy quality was influenced by diagnostic categories, correlated positively with bone marrow fibrosis and negatively with the patient's age. The trephine quality in practice may be lower than the published or declared standards. Ideally the procedure should be executed by the practitioners making more than one trephine a week. Prior to the biopsy it is possible to estimate the level of difficulty posed by an individual patient and use this information to minimize the risk of obtaining an inadequate core.

The authors present a rare case of a synovial sarcoma involving both pleural cavities in a 66-year old woman, confirmed by the t(X;18) translocation detected using the FISH method.

The proposed grading system for malignant fibrous histiocytomas (MFH) comprises 3 grades of malignancy. Analogous to other grading systems, the system includes the factors of mitotic rate and necrosis. In addition to these two factors, the concept of cellularity was included. The prognostic relevance of the grading systems published by Costa, Coindre, van Unnik, Pezzi and Tsujimoto as well as the grading system proposed by the present study was tested on 161 MFH. The results showed that all grading systems tested produced clearly significant differences (p0.01) with regard to the survival estimated for patients with various grades of malignancy. These results revealed the superiority of systems that use 3 grades of malignancy over a 2-grade classification. The proposed grading system yielded a lower percentage of grade II tumours (37%) than the grading systems of Coindre (60%) and van Unnik (70%). In the multivariate analysis of all grading systems, the proposed grading system was the only one to show prognostic relevance (p0. 05).

Twenty-nine cases of oral melanomas were investigated for nm23 and Ki67 antigen expression, as well as for the fraction of tumour cells in S-phase, using immunohistochemical techniques and DNA cytophotometry. Nm23 expression was significantly reduced and Ki67 antigen expression increased in primary tumours with either lymph node or organ metastases in comparison to tumours without metastases. The percentages of Ki67 immunoreactive tumour cells and cells in S-phase correlated positively with each other and negatively with the percentage of nm23-expressing cells. These data argue against a significant growth stimulatory function of the nm23H1 gene product nucleoside diphopshate kinase in the progression of oral melanomas. The functional relevance of nm23 in relation to increased proliferation and metastatic spread is discussed.

Gastrointestinal stromal tumors (GISTs) are common mesenchymal tumors of the gastrointestinal tract. Activating KIT or PDGFRA (platelet-derived growth factor receptor alpha) mutations have been shown to be a major force in GIST pathogenesis. Recently, a previously undescribed N659K PDGFRA exon 14 mutation has been reported in GISTs. The purpose of this study was to evaluate the frequency of GISTs with PDGFRA exon 14 mutations and define the clinicopathologic profile of such tumors. In all, 200 GISTs negative for mutations in KIT exons 9, 11, 13 and 17 and PDGFRA exons 12 and 18 were evaluated for PDGFRA exon 14 mutations by PCR amplification and direct sequencing. Mutations were found in 11 of 119 (9%) gastric GISTs. None of the 81 GISTs from other than gastric location had such a PDGFRA mutation. A majority of these mutations (eight cases) represented simple 2125CA or CG missense mutations, leading to substitution of the lysine for asparagine (N659K). However, in two cases, 2123AT missense mutations leading to substitution of the tyrosine for asparagine (N659Y) was found instead. Of 11 PDGFRA N659-mutant GISTs, 10 had pure epithelioid morphology. One tumor had mixed, predominantly spindle and focally epithelioid cell morphology. Frequency of PDGFRA N659-mutant GISTs among pure epithelioid GISTs was almost 19%. Immunohistochemically, the majority (64%) of these tumors lacked KIT expression or showed only focal scattered KIT positivity. Tumor size ranged from 2.5 to 16 cm (average 7.1 cm). Low mitotic activity,or=5 mitoses/50 high power field was detected in six GISTs including larger,5 cm tumors. Based on mitotic activity and tumor size, six tumors were classified as probably benign with very low malignant potential. Low to moderate malignant potential and high malignant potential was suggested in three and two tumors, respectively. In four cases with moderate or high malignant potential GISTs, a long-term follow-up (average 235.5 months) showed favorable course of disease.

The report presents 200 cases of gastrointestinal stromal tumors (GIST). The material originated from six diagnostic centers in Poland and was reclassified according to the current criteria. Among lesions other than GISTs, 14 were identified as smooth muscle tumors and seven as neural tumors. GISTs were located in the stomach (51-63.3% of the investigated series), small intestine (27.4-33.8%), colon (approximately 4.5%), abdominal cavity, i.e. in the peritoneum and omentum (6%), and in the retroperitoneal space (2.5%). A slight predominance of women was noted (53-56%). The age of the patients ranged between 14 and 93 years of life, with the mean age of 62.4 years. Individuals younger than 45 years of age accounted for 10% of the group. In ten patients (five of them less than 45 years of life), multiple tumors were detected, their number ranging from two to less than 20; these individuals constituted 5% of the entire series. Moderately and highly aggressive tumors predominated. In the series, when multiple tumors were excluded, a total of 24 epithelioid GISTs (12%) were observed; of this number, 13 were situated in the stomach, six--in the small intestine, two--in the abdominal cavity and another two in the retroperitoneal space. Synchronic tumors observed in patients with GISTs were seen in seven patients, including an adenocarcinoma of the colon, two adenocarcinomas of the stomach, a carcinoid tumor of the small intestine, a pheochromocytoma of the retroperitoneal space, an anaplastic lymphoma and a disseminated squamous cell carcinoma. In immunohistochemical reactions (CD117, CD34, SMA, S-100, DES), attention was focused on the immunoreactivity of small GISTs, below 2 cm in size, and of multiple tumors. Immunohistochemical reactions were equally differentiated as to their presence and intensity in small tumors and in highly aggressive lesions above 5-10 cm in size. In multiple GISTs, immunohistochemical tests strongly indicated the heterogeneity of neoplastic cells, which, nevertheless, showed no consistent association with the location of the tumor, its aggressiveness, cellular structure or a tendency to form multiple foci.

Philadelphia-chromosome-negative chronic myeloproliferative disorders (Ph- CMPDs)--essential thrombocythemia (ET), chronic idiopathic myelofibrosis (CIMF), and polycythemia vera (PV)--may show clinical and morphological similarities, particularly at the early stages. The differential diagnosis of Ph- CMPDs is important due to their different treatment and prognosis. Cytological features of megakaryocytes are considered valuable in this differentiation. To establish an objective measure of megakaryocyte dysplasia in Ph- CMPDs, we performed computer-assisted morphometry of more than 4,000 cells from 20 cases of ET, 10 of CIMF, 10 of PV, and 10 controls. Megakaryocyte sets from three Ph- CMPDs differed significantly in respect to many planimetric parameters, but not a single shape or size parameter could have been used as a discriminative tool between the entities. However, the discriminant function analysis with the simultaneous assessment of 12 planimetric variables allowed for a proper classification of 20 of 20 ET, 10 of 10 PV, and 9 of 10 CIMF cases based solely on the morphometric features of megakaryocytes. Additionally, we identified certain new patterns of megakaryocytes specific for ET, PV, and CIMF, which, although not dominating in one Ph- CMPD, are unlikely to occur in two others. Objective measurements of megakaryocyte sizes and shapes may assist the diagnosis of Ph- CMPDs.

Anaplastic lymphoma kinase (ALK)-positive diffuse large B-cell lymphoma (DLBCL) is a rare, recently defined tumor distinct in many aspects from ALK-positive anaplastic large cell lymphoma (ALCL). We present two additional cases of ALK+DLBCL recently diagnosed in our department and a review of literature. A 48-year old man presented with a large upper neck mass growing slowly over 18 months. Histologically the tumor was diagnosed as an ALK-positive diffuse large B-cell lymphoma. with plasmablastic features. Large, frequently intrasinusoidal tumor cells expressed CD138, EMA, weakly IgA and kappa, but were negative for other B-cell markers, T-cell markers and CD30. The ALK staining was cytoplasmic with the increased intensity in the Golgi area. At the diagnosis the patient manifested with the stage IIIB. Three courses of CHOP resulted in partial and only transient remission. The patient died of massive bleeding from his decomposing tumor 3 months after the diagnosis. A 49-year old man complaining of abdominal pain revealed abdominal lymphadenomegaly and a gastric infiltrate, involving the deep portions of the gastric wall. The tumor showed immunoblastic/anaplastic morphology, with some Reed-Sternberg-like cells positive for ALK. ALK immunostaining was cytoplasmic, weak in a routine immunostain, enhanced with double (proteinase + pressure cooker) antigen retrieval. FISH was consistent with the t(2;5)/nucleophosmin(NPM)-ALK rearrangement. The tumor demonstrated similar "null" B/T phenotype with positivity for IgA, lambda, EMA and LCA. The patient (stage IVB) currently undergoes chemotherapy. ALK-positive DLBCL affects mostly middle-aged men, shows generally poor but stage-dependent prognosis (at least 60% mortality rate), presents typically as a lymph node-based disseminated disease, and very rarely involves the bone marrow. Genetic studies showed that the majority of ALK+DLBCL cases are characterized by the clathrin (CLTC)-ALK fusion and in a few cases the NPM-ALK rearrangement has been found.

The aim of this study was to evaluate the influence of suspected prognostic factors on survival in the endemic goiter area. The retrospective review of 208 consecutive patients with differentiated thyroid cancer treated at I Department of General Surgery Collegium Medicum Jagiellonian University from 1983 to 1996. In the studied population there were 98 patients (47.1%) with follicular cancer and 110 patients (52.9%) with papillary cancer. The 182 (87.5%) female and 26 (12.5%) male patients had mean age 48.8 (range from 16 to 80 years). The mean follow-up period was 11.7 years with the longest time of observation 19 years and the shortest 6 years. All the patients were living in the endemic area. Patients with differentiated thyroid cancer have very good prognoses. The 10 year survival rate in case of follicular and papillary cancer were 87.6% and 90.24% respectively. The univariate analysis identified sex, age, histological subtypes (insular and tall cells variants), extrathyroidal extension, size of the tumour beyond 70 mm, lymph node and distance metastases as well as type of surgical procedures in III stage of disease as significant prognostic factors with major effect on survival. In the multivariate analysis extrathyroidal extension, distance metastases, recurrence and lymph node metastases influenced the survival. Early detection of differentiated thyroid cancer with the help of ultrasound examination with aspiration biopsy of suspected lesions, radical surgical procedures and good histological evaluation of the removed tissues guarantee very good treatment results.

The homeobox genes are transcription factors that control the development of tissues and organs. In the colon one of such genes is CDX-2. In colorectal carcinomas, the CDX-2 expression is reduced. The aim of the present study was to investigate the presence of CDX-2 in colorectal carcinomas and to relate it to the histological features and microsatellite stability status. The material consisted of 20 carcinomas without microsatellite instability, 19 cases with low microsatellite instability and 19 cases with high microsatellite instability. CDX-2 expression was investigated using immunohistochemistry with CDX2-88 monoclonal antibody and assessed semiquantitatively. In 10 cases no expression of CDX-2 was observed, while in 6 the protein was present in less than 25% of tumor cells. It was noted that reduced expression of CDX-2 was more frequent in carcinomas situated proximally to the splenic flexure (p0.015) and in tumors with solid growth pattern (p0.03). On the other hand, no significant differences were encountered between groups differing in microsatellite stability. The results suggest that the major factors that determine the presence of CDX-2 in colorectal carcinomas at the protein product level may include cancer location and the solid phenotype of the tumor.

The unclear role of cytokeratin (CK) in the progression and diagnostics of malignant melanomas stimulated us to compare the reactivity of three antibodies directed to CK in 109 paraffin-embedded melanomas. By far the majority of melanomas did not express cytokeratin even at the1% level, only vimentin. In about 6% of melanomas it was possible to find CK expression ranging between 3 and 40% of melanoma cells. There was a correlation between CK expression and pT-stage. Cytokeratin-expressing tumours were found in the more advanced pT-stages. The independent prognostic values of none of the three CK antibodies investigated could be shown.

Melatonin, an antioxidant, protects the pancreas against acute inflammation but, although this indole is released mainly at night, no study has been undertaken to determine circadian changes of plasma melatonin levels and the severity of acute pancreatitis. The aims of this study were: (a) to compare the severity of caerulein-induced pancreatitis (CIP) produced in the rat during the day and at the night, and (b) to assess the changes of plasma melatonin level and the activity of an antioxidative enzyme; superoxide dismutase (SOD), in the pancreas subjected to CIP during the day time and at night without or with administration of exogenous melatonin or its precursor; l-tryptophan. Rats were kept in 12 hr light/dark cycle. CIP was induced by subcutaneous infusion of caerulein (5 microg/kg/hr for 5 hr). Melatonin (5 or 25 mg/kg) or l-tryptophan (50 or 250 mg/kg) was given intraperitoneally 30 min prior to the start of CIP. CIP induced during the day time was confirmed by histological examination and manifested by pancreatic edema, and rises of amylase and lipase plasma activities (by 400 and 500%, respectively), whereas pancreatic SOD, pancreatic blood flow (PBF) and oxygen consumption by pancreatic tissue (VO(2)) were decreased by 70, 40 and 45%, respectively, as compared with the appropriate controls. All morphological and biochemical parameters of CIP induced at night were significantly less severe, compared with those recorded during the light phase. Plasma melatonin immunoreactivity was significantly higher during the night, than during the day, especially following administration of melatonin or its precursor, which reversed all manifestations of CIP. In conclusion, a circadian rhythm modulates the severity of CIP with a decrease of pancreatitis severity during the night compared with that at the day time and this may be due to the increased plasma level of melatonin and higher activity of SOD in the pancreas.

Leptin is a hormone implicated in the regulation of the food intake and body weight, but also increasing number of evidence suggest that leptin participates in the regulation of inflammatory processes. The aim of our study was to examine the influence of exogenous leptin administration on the development and the course of acute ischemic pancreatitis. Acute pancreatitis was induced by temporary limitation of pancreatic blood flow, followed by reperfusion. Leptin was administered three times daily at the dose 10 or 50 micrograms/kg. Studies were terminated at 1, 3, 5, 10 and 21 days after induction of acute pancreatitis. Leptin administration reduced development of pancreatic damage and accelerated pancreatic regeneration. It was manifested by the decrease in serum lipase and amylase activity, the reduction in serum interleukin-1 beta concentration and the improvement of pancreatic histology. Additionally, treatment with leptin caused the increase in the pancreatic blood flow and pancreatic DNA synthesis. Serum interleukin-10 concentration was not effected by leptin administration. Leptin at the dose 50 micrograms/kg was more effective than 10 micrograms/kg. We conclude that leptin is able to limit the pancreatic damage in the course of ischemic pancreatitis and accelerates the pancreatic tissue repair. These effects of leptin seem to be dependent on the increase in pancreatic cell growth, the limitation of pro-inflammatory interleukin-1 beta release and the improvement of pancreatic blood flow.

This study was undertaken to analyze patterns of expression of critical cell cycle regulators (CCR) involved in the p53 pathway in malignant fibrous histiocytomas (MFH). Protein expression was assessed using immunohistochemistry analyzing p53, hMDM2 and p21 (WAF1/CIP1) phenotypes. p53- and hMDM2-positive phenotypes were found to be associated with low p21 levels (p0.01). Positive hMDM2 phenotype did not correlate with any hMDM2 mutations, which in our tumor collective were not found. High-grade MFH differed from MFH grade I and II concerning higher p53 and lower p21 levels, while hMDM2 expression was independent of grade. Inclusion of categorized values into a Cox regression study proved the independent prognostic relevance of p53, hMDM2 and p21 phenotypes.

Chronic myeloproliferative disorders (CMPD) are traditionally diagnosed using criteria based on clinical parameters. A framework for an alternative, trephine bone marrow histology-based approach, was provided by the Hanover group of hematopathologist (the "Hanover classification"). The present study describes a single institution experience with the Philadelphia-BCR/ABL negative CMPD diagnosed on the basis of the histopathology of bone marrow in the three consecutive years (2000 - 2002). Among 246 cases of CMPD (M:F=1:1.6), there were 75 cases of idiopathic myelofibrosis (IMF), 45 of polycythemia vera (PV), 93 of essential thrombocythemia (ET), and 33 cases that were unclassifiable on the basis of morphology (CMPD-U). The clinical profiles of the IMF, PV and ET group emerging from the histological examination correlated with the expected clinical features of these diseases. The ET patients were the youngest (median 51 years) compared to PV (59.5 years), IMF (63.9), and CMPD-U (54.7). In 158 cases (74.3%), the biopsy corroborated the preliminary clinical diagnosis of CMPD, and in the half of these cases it refined the clinical diagnosis of suspected unspecified CMPD placing the disease in a particular specific category (ET, IMF or PV). In the remaining cases the biopsy was done due to an abnormality of unknown origin (usually an accidentally discovered thrombocytosis) or the clinical picture suggesting a disease other than CMPD (11.7%). Some cases of CMPD presented with atypical histological features, such as slight megakaryocytic dysplasia in ET (not justifying the diagnosis of IMF), raising the issue of the subjectivity of histological diagnosis. The trephine bone marrow biopsy provides a useful tool for the diagnosis of CMPD, particularly in the early IMF that may present with a clinical picture undistinguishable from ET, but which carries poorer prognosis and requires more vigorous treatment. A special attention should be paid to the CMPD-U group. Its current nosological status (early phases of IMF/ET/PV or distinct entity or entities?) is still unclear and requires further research.

The blood supply of myomatous uteri collected upon autopsy was examined. The uterine vascular beds were perfused via afferent vessels with fixative followed by Mercox resin and corroded after polymerisation of the resin. The vascular casts thus obtained were examined using scanning electron microscopy. The vascular system of the uterine fibroids was also examined using immunohistochemical analysis (FVIII, factor VIII-related antigen).

Hepatocyte growth factor (HGF) overexpression is observed in experimental and clinical acute pancreatitis. Moreover, previous studies have shown that administration of HGF reduces pancreatic damage in experimental pancreatitis. The aim of our studies was to determine the role of cyclooxygenase-1 and cyclooxygenase-2 in the protective effect of HGF administration against caerulein-induced pancreatitis. Acute pancreatitis was induced in rats by infusion of caerulein. HGF was administered twice at the dose 10 microg/kg s.c. The activity of cyclooxygenase-1 and cyclooxygenase-2 was inhibited by resveratrol and rofecoxib, respectively (10 mg/kg). Immediately after cessation of caerulein or saline infusion, pancreatic blood flow, pancreatic cell proliferation, pancreatic prostaglandin E(2) generation, plasma lipase activity, plasma interleukin-1 beta and interleukin-10 concentration were measured and morphological signs of pancreatitis were examined. Expression of cyclooxygenase-1 and cyclooxygenase-2 mRNA transcripts was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). Cyclooxygenase protein production was analyzed by Western blot. Administration of HGF or caerulein alone, or their combination, was without effect on cyclooxygenase-1 mRNA expression in pancreatic tissue. Expression of cyclooxygenase-2 mRNA was increased by HGF and caerulein. The maximal increase in cyclooxygenase-2 mRNA expression was observed when HGF administration was combined with caerulein infusion. A similar effect was observed when we studied the influence of HGF and caerulein on pancreatic cyclooxygenase-2 production, as determined by Western blot. Administration of HGF without induction of acute pancreatitis increased pancreatic prostaglandin E(2) generation and plasma interleukin-10, and this effect was abolished by the cyclooxygenase-2 inhibitor, rofecoxib. Treatment with HGF, during the development of pancreatitis, increased the plasma interleukin-10 concentration and attenuated pancreatic damage, as evidenced by: (a) histological improvement of pancreatic integrity; (b) the partial reversal of the decrease in DNA synthesis and pancreatic blood flow; (c) the reduction in pancreatitis-evoked increase in plasma lipase and interleukin-1 beta. Administration of resveratrol and rofecoxib alone was without effect on the development of pancreatitis. Combination of rofecoxib with HGF reduced the HGF-evoked increase in plasma interleukin-10 concentration and pancreatic prostaglandin E(2) generation, and abolished the protective effect of HGF against pancreatic damage in pancreatitis. Resveratrol did not affect the protective effect of HGF. We conclude that: (1) HGF induces cyclooxygenase-2 but not cyclooxygenase-1 expression; (2) inhibition of cyclooxygenase-2 in HGF-treated rats decreases the release of anti-inflammatory interleukin-10, increases the production of pro-inflammatory interleukin-1 beta and reduces pancreatic blood flow; (3) cyclooxygenase-2 activity is necessary for the protective effect of HGF in acute pancreatitis.

The objective of the investigation was to compare the degree of interobserver agreement in determining the histological grade of uterine endometrial endometrioid adenocarcinoma using the criteria proposed by the three-grade FIGO classification (1988) and the new, two-grade system proposed by Lax et al. (2000). In the FIGO system, the assessment is focused on the amount of solid, non-squamous growth pattern and the additional feature is the presence of the so-called "notable nuclear atypia" (nuclear grade), with the latter criterion not having been precisely defined. In the two-grade system, the evaluation concentrates on the amount of the solid component, regardless of its character, type of neoplastic growth pattern (expansive or diffusely infiltrating) and the presence of necrosis within the tumor mass. A total of 133 cases of uterine endometrial carcinoma were evaluated, determining the stage according to the FIGO classification and assessing the histological grade based on the criteria presented by the above two systems. All the cases were separately examined by 5 pathologists with varying degrees of experience in gynecological pathology. A higher degree of interobserver agreement was demonstrated when the two-grade system was employed as compared to the FIGO system, regardless whether the material was evaluated by experienced pathologists (FIGO k - 0.64 - 0.71, binary - 0.91 - 0.92), or by individuals with little experience in gynecological pathology (FIGO k - 0.23 - 0.48, binary - 0.21 - 0.57). The data point to the superior character of the two-grade system as to the agreement of the histological grade assessment, but also suggest a considerable effect of experience on the precision of the evaluation.

Defects in DNA mismatch-repair genes MLH1 and MSH2 reported primarily in hereditary nonpolyposis colorectal carcinoma are present in many sporadic tumors, including malignant melanomas. The main aim of this study was to investigate the expression of these genes in malignant melanomas in relation to tumor stage. An experiment was performed on paraffin-embedded tissue microarrays of malignant melanomas applying in situ hybridization with probes produced by our research group and immunohistochemical techniques. In situ hybridization demonstrated MLH1 expression in 45 of 59 melanomas and MSH2 expression in 51 of 59 melanomas. Immunohistochemistry detected MLH1 expression in 46 of 59 melanomas and MSH2 expression in 50 of 59 melanomas. Down-regulation of expression of both DNA mismatch repair genes in malignant melanomas was observed. The findings obtained by in situ hybridization and immunohistochemistry correlated significantly. Our study demonstrates the suitability of in situ hybridization with MLH1 and MSH2 probes for paraffin-embedded tissue. Tissue microarrays can be used successfully in both in situ hybridization and immunohistochemistry to analyze the expression of DNA mismatch-repair genes.

Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract. GISTs express KIT and show gain-of-function KIT mutations. Most of these mutations affect the KIT juxtamembrane domain, but other KIT domains are mutated at a lower frequency. In this study, frequency of GCC TAT insertion mutation (1530ins6) in KIT exon 9 (extracellular domain) and its possible clinicopathologic significance was investigated. Screening of 520 GISTs identified 26 cases with 1530ins6 KIT mutation and confirmed the previously reported low frequency of this type of KIT mutation among GISTs of different locations. Of the 26 tumors with 1530ins6 KIT mutation studied, 21 originated from the small intestine, 1 from the colon, and 3 from the rectum. In 1 case, primary small intestinal versus colonic localization could not be clearly established because of intra-abdominal dissemination. No distinctive morphological features were identified for the cohort of tumors defined by 1530ins6 KIT mutations. Most of the tumors showed predominant spindle cell morphology, and a few cases had epithelioid or pleomorphic histological features. Following previously published criteria based on tumor size and mitotic rate, 22 of 26 (85%) tumors were classified as malignant or potentially malignant, and 4 (15%) were classified as probably benign. A malignant clinical course was documented in 18 of 19 tumors from the malignant category. The survival times of 11 patients who died of disseminated GISTs ranged from 1 month to 105 months (median survival time, 26 months). In contrast, 2 of 4 GISTs assigned as probably benign tumors with follow-up information had long disease-free survival. GISTs carrying 1530ins6 occur exclusively in the intestinal location, and a great majority of these tumors follow a malignant course.

Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract. GISTs express KIT and have KIT mutations. Majority of these mutations cluster in the 5' end of the KIT juxtamembrane domain. Little is known about the clinicopathological profile of GIST carrying internal tandem duplications in the 3' end of KIT juxtamembrane domain (ITDs in the 3' KIT-JM). In this study, 500 immunohistochemically KIT-positive GISTs were screened for this type of mutation, and 18 cases were identified (3.6%). The majority of the ITDs consisted of 1 to 18 codon duplications, with Tyr(578), Asp(579), and Leu(576) being the most commonly duplicated codons. There were 14 gastric (78%), 2 small intestinal (11%), and 2 anal (11%) primary tumors diagnosed in 12 females and 6 males with median age of 71 years. The frequency of IDTs in gastric GISTs was 6.5% and was only 0.5% in intestinal GISTs. There was a strong female predominance (79%) among the patients with gastric tumors. Histologically, 16 GISTs were spindle cell, and 2 had epithelioid morphology. The sizes of primary tumors varied from 1 to20 cm. Based on the combination of tumor size and mitotic activity, six tumors were classified as benign or probably benign, eight as having uncertain malignant potential, and only four as malignant. Follow-up data available in 17 patients confirmed the malignant course of disease in 3 cases. Only one of the tumors classified as potentially malignant metastasized, although the follow-up was limited in some cases. In summary, the great majority of GISTs with ITDs in the 3' KIT-JM were mitotically inactive tumors occurring predominantly in the stomach and that seemed to have a favorable course. This suggests that presence of these IDTs may define a clinicopathologically favorable subset of GISTs. The consequence of these mutations to KIT signaling should be investigated.

Insulin-like growth factor-I (IGF-I) is regarded as one of mammary tissue proliferative factors. Insulin-like growth factor binding protein-3 (IGFBP-3) limits the IGF-I binding potential to its receptor. That limits the IGF-I bioavailability. Recently experimental studies indicated that insulin-like growth factor binding proteins (IGFBPs) might have their own biological actions beyond their ability to regulate insulin-like growth factors (IGFs). Our earlier results showed the progesterone-induced rise in hGH and IGF secretion by human breast cancer explants.To determine the ability of progesterone to stimulate simultaneous local IGF-I and IGFBP-3 secretion by non-malignant and malignant mammary tissue collected from different receptor phenotype tumours.Explants from the tumour and surrounding normal non-malignant tissue were obtained during surging. Breast cancer explants were defined as: ER+ PR+; ER-PR-; ER+ PR-; and ER-PR+. Part of the explants was fixed in 10% buffered formalin for steroid receptor determination by immunohistochemistry. Other parts were cut into small pieces, weight and cultured in Parker medium (M199) supplemented with 5% of calf serum at 37 degrees C in an atmosphere containing 5% CO2 for 48 hours in control medium or with the addition of progesterone (10-7 M). Later media were collected for IGF-I and IGFBP-3 concentration analysis.Progesterone increased (p0.01) IGFBP-3/IGF-I index in ER(-)PR(-) non-malignant tissue and decreased the IGFBP-3/IGF-I index in ER(-)PR(+), ER(+)PR(-) non-malignant explants. That increased the IGF-I bioavailability. Breast malignant explants showed the progesterone induced IGFBP-3/IGF-I index decrease. The decrease was most evident (p0.01) in malignant explants expressing progesterone receptor.Progesterone increased local IGF-I bioavailability in malignant breast tissue. That phenomenon depended on steroid receptor phenotype of breast tissue and was most evident in tissue expressing progesterone receptor. In non-malignant tissue that phenomenon was also found in estrogen receptor expressing tissue. Lack of steroid receptor expression in breast explants reversed that phenomenon.

Previous studies have shown that sensory nerves and calcitonin gene-related peptide (CGRP) affect caerulein-induced pancreatitis. The aim of this study was to examine the role of capsaicin-sensitive nerves and the impact of CGRP administration on necrotizing pancreatitis induced by ischemia/reperfusion.Ablation of sensory nerves was made by capsaicin 10 days before induction of pancreatitis. Acute pancreatitis was induced in rats by limitation of pancreatic blood flow (PBF) followed by reperfusion. Treatment with saline or CGRP (10 g/kg s.c.) or stimulation of sensory nerves by low doses of capsaicin (0.5 mg/kg s.c.) was performed 1 h before ischemia. After 1 h reperfusion we examined pancreatic blood flow (PBF), plasma amylase and lipase activity, plasma interleukin-1beta (IL-1beta) concentration, pancreatic DNA synthesis and morphological signs of pancreatitis.Ischemia followed by 1 h reperfusion led to induction of necrotizing pancreatitis, manifested by morphological signs of pancreatic damage, decrease in pancreatic DNA synthesis and PBF, as well as an increase in plasma amylase and lipase activity and plasma IL-1beta concentration. Both, treatment with CGRP and stimulation of sensory nerves attenuated pancreatic damage. Ablation of sensory nerves enhanced I/R evoked pancreatic damage. The deleterious effect of deactivation of sensory nerves on I/R-induced pancreatitis was partly reversed by administration of CGRP prior to I/R.Stimulation of sensory nerves protects the pancreas against damage evoked by I/R, whereas ablation of these nerves aggravates tissue damage in the pancreas exposed to I/R. The beneficial effect of sensory nerves is partly dependent on CGRP release.

Epstein-Barr virus plays a substantial role in numerous human neoplasms including gastric carcinoma, as proved recently. In our series EBV has been detected in five cases of forty (12.5%) gastric carcinomas. It may indicate that Poland is one of the countries with the highest EBV-induced gastric carcinoma incidence worldwide.

A significant percentage of conventional schwannomas, whether sporadic or associated with neurofibromatosis 2 (NF2), show loss of heterozygosity (LOH) at NF2 and/or NF2 inactivating mutations. Similarly, a significant percentage of neurofibromas show LOH at NF1 and/or NF1 inactivating mutations. There are no molecular genetic data on gastrointestinal (GI) nerve sheath tumors traditionally diagnosed as benign schwannomas, rare neoplasms possibly derived from the schwannian elements dispersed between the smooth muscle fibers. In this study, we analyzed 1 esophageal, 16 gastric, 1 small intestinal, and 2 colonic tumors of such type. Histologically, all were spindle cell neoplasms positive for S-100 protein, vimentin, and glial fibrillary acidic protein, and negative for smooth muscle markers, KIT, CD34, neurofilament proteins, and HMB45. Focal or extensive lymphoid cuffs, often containing germinal centers, were present in most cases. None of the patients had NF2 or NF1. Chromosomes 22 and 17, particularly NF2 and NF1 loci, were analyzed for LOH in all GI tumors and for comparative purposes in 10 conventional schwannomas. LOH on 22q was seen in 40% of conventional schwannomas but in only 5% (1 of 20) of GI schwannomas. PCR amplification followed by direct sequencing of PCR products failed to identify mutations in NF2 coding sequences (exons 1-15) in 13 cases, including a case with LOH on 22q. Losses on 17q involving NF1 were seen in both GI and conventional schwannomas in 50% and 33% of analyzed tumors, respectively. LOH at NF1 might be one of the genetic features seen in peripheral nerve sheath tumors from different locations and should be interpreted with caution. However, lack of NF2 alterations strongly supports the hypothesis that GI schwannomas represent a morphologically and genetically distinct group of peripheral nerve sheath tumors that are different from conventional schwannomas.

To analyze cytologic and histologic parameters in bone marrow trephine biopsy in an attempt to define heterogeneity of hairy cell leukemia cells.The study group consisted of 28 trephine biopsies. Immunohistochemistry for CD20 antigen was used. Image processing and measurements were performed with AnalySIS 3.0 image analysis system (Soft Imaging System GmbH, Germany) and custom built programs. For planimetric measurements of nuclei, automatic segmentation was implemented. The measured parameters were: surface area, perimeter, minimum, mean and maximum diameter, and a set of form factors. Relative volumes of bone trabeculae, adipose tissue, hematopoietic tissue and neoplastic infiltrate were assessed by the point counting method. Nuclear volume was measured by the point sampled intercept method. Bone marrow fibrosis was assessed using a curvilinear line test system.Significant variability of cell nuclei was found, and their classification into 3 types was possible. The relative frequency of those types was different in various cases and allowed subdivision of cases into 3 groups that differed in some clinical and histologic manifestations.The present study demonstrated the heterogeneity of cell populations of hairy cell leukemia.

Application of acrylic emulsion Liquitex R for injection studies of the vascular system of human myomatous uteri was analysed. It was found that this injection mass does not penetrate the capillary bed of human organs, but it is useful in studies carried out on the blood supply of the human organs removed from cadavers. The results were compared with the studies performed with the help of immunohistochemical tests for von Willebrandt's factor.

DNA mismatch repair system defects cause microsatellite instability (MSI) and form an alternative pathway in cancer development. Germline mutations of DNA mismatch repair genes account for hereditary nonpolyposis colorectal cancer, which has a different morphology and biology than sporadic cancers. MSI has also been found in sporadic neoplasms and some inflammatory conditions (chronic pancreatitis, ulcerative colitis). The purpose of the present study was to evaluate the expression of hMLH1 and hMLH2 proteins in infiltrating pancreatic cancer and to find out whether there is a relationship between some phenotypic manifestations and expression of MMR genes. We studied 30 cases of infiltrating pancreatic cancer and apart from hMLH1 and hMLH2 expression cytokeratin 7 and chromogranin were measured as markers of ductal and endocrine differentiation, respectively. All ductal pancreatic cancers expressed cytokeratin 7. In most cases the expression was strong, present in 50-100% of cells in moderately differentiated cancers and in 80-100% of cells in poorly differentiated cancers. Chromogranin expression was seen in 5 moderately differentiated cancers and in 6 poorly differentiated cancers (up to 20% of positive cells). In all cases DNA mismatch repair genes expression was present.Ductal pancreatic carcinomas express hMLH1 and hMLH2 proteins irrespective of their differentiation. The expression of cytokeratin 7 is typical of ductal pancreatic carcinoma and its level is related to cancer differentiation. Some ductal pancreatic carcinomas irrespective of their differentiation show the expression of chromogranin, which is associated with the expression of hMSH2 gene.

Background Helicobacter pylori infection of Mongolian gerbils is an established model of gastric carcinogenesis, but gastric secretory aspects of this carcinogenesis have not been studied. Methods The effects of single intragastric inoculation of gerbils with H. pylori strain (cagA+ vacA+, 5 x 10(6) CFU/ml) or vehicle (saline) were examined at 1, 2, 4, 6, 9, 12 and 30 weeks from inoculation. Gastric morphology, the presence of H. pylori using the rapid urease test, the density of H. pylori and 16S rRNA and the plasma gastrin and somatostatin were determined. Results H. pylori was detected in gastric mucosa in all infected animals. Basal gastric acid in gerbils was reduced by about 50% after H. pylori inoculation. Early lesions seen at 4 weeks after H. pylori inoculation consisted of chronic gastritis with thickened mucosal folds, oedema, congestion and mucosal lymphocytic infiltration. Adenomatous hyperplasia with cellular atypia with increased mitotic activity and the formation of apoptotic bodies and visible erosions and ulcerations were observed at 12-30 weeks after inoculation. The atypical gastric glands were situated 'back-to-back', suggesting gastric pre-cancer. The gastric blood flow in H. pylori-infected gerbils was significantly lower than that in the controls. Six- to seven-fold increase in plasma gastrin levels combined with significant fall in gastric somatostatin contents and the intraepithelial neoplasia were noticed in gerbils at all tested periods. Conclusion H. pylori-infection in gerbils resulted in gastric pre-cancer associated with functional changes, such as suppression of gastric secretion and impairment of both gastric mucosal microcirculation and the gastrin-somatostatin link.

A case of the coexistence of nonfunctional pituitary adenoma and pulmonary sarcoidosis is reported. 39 years old female presented symptoms of a pituitary-gonadal axis insufficiency, visual deficit and dizziness. CT pituitary imaging revealed large intra and extrasellar tumour. Histological examination of the tissue obtained at transsphenoidal surgery showed chromophobic adenoma. Hypopituitarism and transient diabetes insipidus occurred after the surgery. The adequate replacement therapy with hydrocortisone and sex steroids was introduced. At the time of the pituitary tumour diagnosis enlargement of pulmonary lymph nodes was also observed. Based on histological examination of tracheal and bronchial epithelium specimens obtained during bronchoscopy the diagnosis of pulmonary sarcoidosis was made. No other systemic sarcoidosis localisation was confirmed. Histological re-evaluation of adenoma showed no noncaseating granuloma tissue. The overlapping symptoms of pituitary adenoma and other intrasellar masses may result in diagnostic difficulties, particularly in the presence of systemic disorders in which this gland may be involved.

The function of the sodium iodide symporter (Na(+)/I(-), (NIS), a membrane protein that mediates iodide transport into cells, is the best described in the thyroid cells. NIS is also found in mammary cells during lactation and in breast carcinoma cells. The aim of this study was evaluation of incidence and grade of NIS expression in invasive ductal breast cancer. Immunohistochemistry using a panel of antibodies against NIS was carried out in surgical paraffin-embedded tissue obtained from 50 patients with invasive ductal breast carcinoma. NIS expression was found in 45 (90%) cases. The demonstration of NIS expression in breast carcinoma cells may provide a novel approach to its diagnosis and treatment.

H. pylori (Hp) -induced atrophic gastritis is a well-known risk factor for the development of gastric cancer. Whether Hp eradication can prevent or retard the progress of atrophy and metaplasia has been the topic of numerous studies but the subject remains controversial. Recently, the increased expression of ornithine decarboxylase (ODC), gastrin and cyclooxygenase (COX)-2 has been shown to be increased in premalignant lesions in gastric mucosa and to play an essential role in the malignant transformation. The aim of the study is to assess the effect of eradication therapy on atrophic gastritis and analyze the gene expression for ODC, COX-2 and gastrin in gastric mucosa after succesful eradication in patients with atrophic gastritis. Twenty patients with chronic atrophic gastritis including both corpus and antrum of the stomach were included in this study. Four antral mucosal biopsy specimens were obtained from antrum and four from corpus. The histopathologic evaluation of gastritis was based on Sydney classification of gastritis. All patients were Hp positive based on the [13C] urea breath test (UBT) and the presence of anti-Hp IgG and anti-CagA-antibodies detected by ELISA. The patients were then eradicated with triple therapy consiting of omeprazol (2 x 20 mg), amoxycillin (2 x 1 g) and clarithromycin (2 x 500 mg) for seven days and vitamin C 1 g/day for three months. In gastric mucosal samples obtained from the antrum and corpus before and after eradication, the mRNA expression for ODC, COX-2, and gastrin was assessed by reverse-transcription polymerase chain reaction (RT-PCR). In all patients the gastric secretory analysis was performed by measuring gastric acid output and serum gastrin levels. After triple therapy the successful eradication assessed by UBT was observed in 95% of patients. In 45% of patients the infection with CagA-positive Hp strain was observed. Three months after eradication a significant reduction in the gastric activity (neutrophilic infiltrate) and severity (mononuclear infiltrate) of gastritis was observed. The atrophy score improved in both antrum and corpus after eradication. The expression of COX-2 and ODC was significantly up-regulated in the gastric mucosa of patients with atrophic gastritis and significantly reduced after eradication therapy. In all successfully eradicated patients with atrophic gastritis a significant increase in gastric acid secretion and decrease in serum gastrin were observed. We conclude that: (1) Hp eradication leads to the decrease in ODC and COX-2 gene expression in the gastric mucosa, and this may be relevant for the prevention of the Hp-associated gastric carcinogenesis; and (2) gastric atrophy ameliorates upon successful Hp eradication therapy.

The paper presents start-of-art knowledge of primary gastric B-cell lymphomas. Despite the progress of the investigations, a lot of questions remain still unanswered and many doubts wait for explanation.