Expression of selected microRNAs in pancreatic ductal adenocarcinoma: is there a relation to tumor morphology, progression and patient's outcome?

Pancreatic ductal adenocarcinoma (PDAC) remains a disease with extremely poor prognosis and limited effective available treatment. Differential expression of miRNAs isolated from tumor tissue has been proposed as a marker for tumor diagnosis, progression, and prognosis. Nevertheless, the prognostic value of miRNAs expression in PDACs for patient outcome still remains unclear. Expression of 7 selected miRNAs, isolated from FFPE samples of 54 PDAC patients, was quantified using RT-qPCR. The relationship of miRNA expression levels with tumor histology, clinicopathological characteristics, patient overall survival (OS), and progress-free survival (PFS), was subsequently evaluated. Overexpression of miR-21, miR-155, and miR-210 was observed in PDACs (up to 72.62, 232.36, and 181.38-fold, respectively), in comparison with non-neoplastic tissues. On the other hand, miR-96 and miR-217 were significantly downregulated in PDACs (up to one hundred times). No differences were, however, noticed between cancer and normal tissues for the expression levels of miR-148a and miR-196a. On the other hand, expression levels of all 7 miRNAs failed to demonstrate a significant correlation with parameters of tumor progression, such as tumor stage, grade, nodal involvement, perineural, and vascular invasion. The positive correlation of miR-210 levels was, however, observed with patient age (ρ=0.35). Additionally, miR-148a and miR-217 expressions have shown a positive association with tubular tumor growth pattern (ρ=0.39; ρ=0.28). The negative correlation of miR-148a values was also demonstrated with dissociative growth pattern and nuclear atypia (ρ=-0.30; ρ=-0.27). Finally, no statistically significant correlation could be demonstrated with the expression levels of all 7 tested miRNAs and PDAC patient survival; neither for OS nor for PFS (p>0.05). Our data have confirmed abnormal miRNAs expression in PDACs in comparison with adjacent non-neoplastic tissue. On the other hand, no correlation was discovered between miRNA expression and parameters of tumor progression. We have found a significant association between histologic tumor growth patterns and miRNA expression, making this work the first study, which analyses this aspect of PDAC. Finally, in our group of patients, no relationship of miRNA levels and patient prognosis could be demonstrated. Therefore, further investigation is required to evaluate the predictive and prognostic potential of miRNAs in a clinical setting.

[Synchronous colorectal carcinoma and non-Hodgkin lymphoma - two case reports]. / Synchronní kolorektální karcinom a nehodgkinský lymfom - popis dvou prípadu.

Colorectal carcinoma represents an important cause of morbidity and mortality in adults, and its incidence in the Czech Republic is one of the world´s highest. The basic therapeutic approach is surgery: surgical removal of the affected part of the bowel together with regional lymph nodes dissection. The lymph nodes are routinely examined by means of histopathology. In this paper, we present two patients whose histological examination of mesocolic lymph nodes revealed an infiltration by synchronous malignant B-non-Hodgkin-lymphoma. Mantle cell lymphoma was present in the first case, and small cell lymphoma CLL/SLL in the other. Relevant literature is reviewed. KEY WORDS: synchronous - malignancy - colorectal - lymphoma - lymph node.

Large contribution of human papillomavirus in vaginal neoplastic lesions: a worldwide study in 597 samples.

AIM: This work describes the human papillomavirus (HPV) prevalence and the HPV type distribution in a large series of vaginal intraepithelial neoplasia (VAIN) grades 2/3 and vaginal cancer worldwide. METHODS: We analysed 189 VAIN 2/3 and 408 invasive vaginal cancer cases collected from 31 countries from 1986 to 2011. After histopathological evaluation of sectioned formalin-fixed paraffin-embedded samples, HPV DNA detection and typing was performed using the SPF-10/DNA enzyme immunoassay (DEIA)/LiPA25 system (version 1). A subset of 146 vaginal cancers was tested for p16(INK4a) expression, a cellular surrogate marker for HPV transformation. Prevalence ratios were estimated using multivariate Poisson regression with robust variance. RESULTS: HPV DNA was detected in 74% (95% confidence interval (CI): 70-78%) of invasive cancers and in 96% (95% CI: 92-98%) of VAIN 2/3. Among cancers, the highest detection rates were observed in warty-basaloid subtype of squamous cell carcinomas, and in younger ages. Concerning the type-specific distribution, HPV16 was the most frequently type detected in both precancerous and cancerous lesions (59%). p16(INK4a) overexpression was found in 87% of HPV DNA positive vaginal cancer cases. CONCLUSIONS: HPV was identified in a large proportion of invasive vaginal cancers and in almost all VAIN 2/3. HPV16 was the most common type detected. A large impact in the reduction of the burden of vaginal neoplastic lesions is expected among vaccinated cohorts.

[Neuroendocrine tumours of the alimentary tract--history and at present]. / Neuroendokrinní nádory trávicího ústrojí--historie a soucasnost.

Histological classification of the neuroendocrine tumours ("carcinoids") of the alimentary tract, as well as the opinion on biological behaviour of these tumours, changed rapidly within the last decade. Advances in knowledge of cellular biology of the diffuse endocrine system and in clinical diagnostics and treatment of tumours lead to the creation of a new histological classification of neuroendocrine tumours. This classification, apart from essential histological picture and immunohistological characterisation of the markers of neuroendocrine differentiation, also includes definition of biological properties of tumours based on their site of origin, mitotic and proliferative activity of the tumour cells and clinicopathological correlation, including the size of the tumour and its progression. Exact classification of an individual tumour into a corresponding category is an essential condition to select adequate following diagnostic procedures and optimal therapeutic strategy.

[Merkel cell carcinoma--immunohistochemical study in a group of 11 patients]. / Karcinom z Merkelových bunek--imunohistologická studie v souboru 11 pacientu.

The aim of our work was to confirm an immunohistochemical profile of routine markers of epithelial and neuroendocrine differentiation in eleven cases of Merkel cell carcinoma, as well as to study the expression of two markers of early phases of neuronal differentiation, namely reelin and class III beta-tubulin, markers which have not yet been studied in Merkel cell carcinomas. In all the investigated tumours the characteristic "dot-like" pattern of cytokeratin 20 immunoexpression, as well as negative immunostaining for cytokeratin 7 and thyroid transcription factor 1 (TTF-1) were disclosed; all the tumours showed neuroendocrine differentiation, expressing either neuron specific enolase (NSE) or chromogranin A(CgA), or both. An interesting finding was observed when the anti-cytokeratin monoclonal antibody MNF 116 was used. The characteristic "dot-like" pattern was detected in high proportion of tumours, including two samples of local recurrence of one of the carcinomas, where neoplastic cells have lost the expression of cytokeratin 20. The majority (91%) of Merkel cell carcinomas included in our group showed positive immunodetection of class III beta-tubulin when TU-20 antibody was used, while TuJ-1 immunostaining was surprisingly negative in all the investigated tumours. Detection of reelin was negative in almost all the studied Merkel cell carcinomas except for cases, where neoplastic cells revealed weak focal immunostaining in a minor portion of neoplastic cells.

Expression of class III beta-tubulin in colorectal carcinomas: an immunohistochemical study using TU-20 & TuJ-1 antibody.

BACKGROUND & OBJECTIVE: Expression of class III beta-tubulin represents newly discovered marker of resistance to taxol-based chemotherapy in a wide spectra of carcinomas. However, very little is known about its expression in colorectal carcinomas. This study was done to determine class III beta-tubulin expression in a large series of colonic carcinomas, covering tumours with different degree of differentiation in order to evaluate its prospective significance in resistance to taxol-based chemotherapeutics and to compare the immunostaining profile of two widely used monoclonal antibodies, TU-20 and TuJ-1 METHODS: Sixty patients with colorectal carcinoma were enrolled; all of them were treated surgically by the resection. Twenty tumours were histologically assessed as G1, 20 as G2 and 20 as G3. Routine immunohistochemical procedure using TU-20 and TuJ-1 mouse monoclonal antibodies was applied to all 60 specimen and slides were evaluated using an optical microscope. RESULTS: Expression of class III beta-tubulin was detected in 14 tumours (23.3%), while remaining tumours were negative. Relatively higher frequency of class III beta-tubulin expression was observed in G3 tumours (10 cases) in comparison with G1 (3 cases) and G2 (1 case), respectively. Seven tumours displayed positive immunostaining with both tested antibodies TU-20 and TuJ-1. Six tumours showed expression of class III beta- tubulin in more than 1 per cent of neoplastic cell population. In remaining 8 tumours only individual scattered neoplastic cells exhibited class III beta-tubulin expression either with TU-20, or with TuJ-1 antibody. INTERPRETATION & CONCLUSION: Higher frequency of immunoreactivity was observed in poorly differentiated tumours. However, more than 90 per cent of neoplastic cell population did not express class III beta-tubulin in almost all tumours. These negative cells of colonic cancer could represent the potential target for taxane-based chemotherapy in the future. Our results indicate that TU-20 and TuJ-1 antibodies exhibit very similar immunoreactivity in neoplastic tissue.

[Leiomyoma of the abdomen wall. Case report]. / Leiomyom piední steny birisní.

Authors present the case report of the patient indicated to plastic of abdomen wall for large hernia. Per operation, tumor was found without communication with abdomen cavity and organs. Histopathologically, leiomyoma of the abdomen wall was identified.

Expansion of human mesothelial progenitor cells in a longterm three-dimensional organotypic culture of Processus vaginalis peritonei.

A 3D culture system was used to investigate the behaviour of mesothelial cells present in the wall of human processus vaginalis peritonei. Small tissue fragments placed on collagen sponges were cultured for 7, 14 and 21 days in medium supplemented with 10% FBS, and analysed for the expression and distribution of cytokeratins (CKAE1-AE3, CK19), p63, Ki-67, vimentin, CD34, and HBME-1. Before culture, flat mesothelial cells displayed immunoreactivity for cytokeratins, vimentin and HBME-1, while p63 and CD34 were negative. Mesenchymal cells within the stroma were vimentin-positive and endothelial cells of small vessels displayed positive staining for CD34. Cytokeratins, p63 and HBME-1 were negative in all stromal cells. In cultured fragments, flat mesothelial cells positive for vimentin, cytokeratins and HBME-1 proliferated, lining the fragment surface and migrating into the sponge. Capillaries showed morphological alterations; however, their immunoreactivity was comparable with the stroma prior to culture. Cells that had migrated into the sponge and displayed characteristics of mesothelial progenitors, predominantly spindleshaped and stellate, showed heterogeneous expression of markers especially in late phases of cultivation. These cells were constantly positive for vimentin, a small fraction was cytokeratin-positive and a few displayed HBME-1 immunoreactivity. CD34 was found in cells forming small cavities into the matrix, resembling newly formed blood vessels. Cells that had migrated into the sponge could be isolated and expanded in coculture with feeder NIH.3T3 fibroblasts. This system is suitable for studying growth and behaviour of mesothelial cells within their natural environment, providing a good method for isolation and expansion of their progenitor cells.

Human papillomavirus infection and tumours of the anal canal: correlation of histology, PCR detection in paraffin sections and serology.

Human papillomavirus infection is an important etiological factor in squamous cell carcinoma of the anus (SCCA). Different histological variants of anal carcinomas displaying squamous differentiation, previously classified as separate tumours, were recently reclassified as SCCA by the WHO. In our recent study the presence of HPV was detected by PCR in biopsy specimens of 42 different anal tumours, including SCCA and its histological variants (n=22), adenocarcinomas (n=5), tubulovillous adenomas (n=5) and anal condylomas (n=10). HR HPV16 (high risk - HR) was detected in 18 of SCCA specimens (81.8%). All histological variants, i.e. tumours with basaloid, squamous and mixed histological patterns, were represented among the HPV-positive cancers. Four tumours (18.2%) were HPV negative. Low-risk (LR) HPV types were not detected within the SCCA group. HPV16 was identified in one adenocarcinoma, while four cases were HPV negative. Two adenomas showed presence of HPV16; one showed simultaneous positivity for HPV33. The remaining three tumours were HPV negative. Seven anal condylomas (70%) were LR HPV 6 and/or 11 positive, while three were HPV negative. The presence of HR HPV types was not observed in anal condylomas. Our results provide further evidence in support of the etiological role of HR HPV infection in the development of SCCA regardless of its histological appearance.

Accuracy of angiography and Doppler ultrasonography in the detection of carotid stenosis: a histopathological study of 123 cases.

BACKGROUND: A prospective study was performed comparing the accuracy of digital subtraction angiography (DSA) and Doppler ultrasonography (DUS) stenosis findings with measurements on histological specimens. METHOD: DSA and DUS were used to evaluate carotid stenosis and were compared with measurements on histological specimens. Intact carotid plaques from 123 cases were removed in one piece during surgery. The specimens were histologically processed and examined in transverse sections. The smallest inner and correlating outer diameters were measured and the extent of stenosis was calculated. Carotid artery stenoses were compared and statistics done. Specimens in symptomatic cases were divided into 3 groups: stenosis 30-49% (Group 1), stenosis 50-69% (Group 2) and stenosis 70-99% (Group 3). Specimens in asymptomatic cases were divided into two groups: stenosisor=60% (Group B). FINDINGS: Wilcoxon paired tests revealed significant differences between DSA, DUS and measurements on histological specimens. In severe stenoses only, no significant difference was observed between stenosis measurement according to the European Carotid Surgery Trial (ECST) angiography methodology and measurements on histological specimens. The most pronounced differences were found between angiography methodology of the North American Symptomatic Carotid Endarterectomy Trial (NASCET) and measurements on histological specimens. When investigating how often preoperative measurement classified stenosis into the same Group of stenoses as postoperative measurement, DUS was the most accurate diagnostic tool. CONCLUSIONS: This study confirmed our previous results, i.e., angiography underestimates the degree of carotid artery stenosis. DUS seems to be more accurate in classifying stenoses into different groups to the extent of narrowing of the carotid arteries. These results make the position of angiography in diagnostic algorithm of carotid stenoses investigations even more questionable.

Expression of CD44v6 correlates with cell proliferation and cellular atypia in urothelial carcinoma cell lines 5637 and HT1197.

CD44 comprises a family of membrane adhesion molecules encoded by a single gene and diversified by alternative splicing and extensive posttranslational modifications. Alterations of CD44 expression patterns are linked to tumour invasion and formation of metastases. However, CD44 expression and its relation to the biological properties of tumours vary depending on the tumour type and origin. In transitional cell carcinoma of the urinary bladder, low CD44 expression is linked to enhanced tumour aggressiveness. We studied CD44 expression in two urothelial cancer cell lines, HT1197 and 5637. CD44s and a v6 variable exon-containing splice variants were detected in both cell lines by reverse transcription-PCR and by commercially available monoclonal antibodies. In both cell lines, Western blot analysis detected immunoreactive proteins with approximate sizes 70-85 kD, 95-110 kD, and 120-140 kD with CD44v6 antibody and weak bands with size 70-98 kD with CD44s antibody. At the cellular level, the pattern of CD44 immunoreactivity correlated with a lower level of cell differentiation and a higher degree of cell proliferation. In HT1197 cells, the CD44v6 was detected predominantly in small proliferating cells and in large multinuclear atypical cells. CD44s and CD44v6 displayed low immunoreactivity in HT1197 cells with a higher degree of epithelial differentiation. The 5637 cells expressed CD44v6 strongly and CD44s weakly. We conclude that CD44v6 expression correlates with a higher proliferative activity and with a stem cell-like phenotype in both cell lines and with cellular atypia in HT1197 cells.

Chromophobe renal cell carcinoma with microcystic and adenomatous arrangement and pigmentation--a diagnostic pitfall. Morphological, immunohistochemical, ultrastructural and molecular genetic report of 20 cases.

We present clinical, morphological, immunohistochemical, ultrastructural and molecular genetic features of 20 cases of a peculiar form of chromophobe renal cell carcinoma (CRCC) with morphology differing from that of conventional CRCC. Microscopically, the typical features of the tumors were microcystic arrangement and formation of adenomatous structures. Microcystic areas were composed of smaller eosinophilic and bigger pale cells having cytological appearance typical of conventional CRCC. Cytological features of the adenomatous structures were mostly different from those of conventional CRCC. They had a typical columnar arrangement with nuclei positioned at the base of the glandular structures and a small amount of a deeply eosinophilic cytoplasm often endowed with brush border facing the lumen of the glands. In addition, all the tumors showed a brown pigmentation. The pigmentation was located mostly extracellularly, where it formed pools of heavy deposits. Microscopic calcifications present in all cases formed psammoma bodies or else the calcifications were more extensive and amorphous in shape. Ultrastructurally, the cells showed features characteristic of CRCC: typical cytoplasmic vesicles were 100-700 nm in size and mitochondria had tubulovesicular, lamellar or circular cristae. Some tumor cells contained dark, variously sized electron-dense pigment granules. Neither melanosomes nor membrane-bound neurosecretory granules were seen. Using fluorescence in-situ hybridization probes for chromosomes 1, 2, 6, 10, 13, 17 and 21, the tumors revealed massive loss of tested chromosomes typical for conventional CRCC. Monosomy of chromosomes 1, 2, 6, 10, 13 and 21 was found in 100, 36, 91, 82, 82, 82 and 64% of cases, respectively. None of the cases showed mutation of exons 9, 11, 13 and 17 of the c-kit gene. The important feature of pigmented microcystic chromophobe renal cell carcinoma is a relatively benign biological behavior and the absence of distant metastases and sarcomatoid transformation.

Cystic fibrohistiocytic tumours presenting in the lung: primary or metastatic disease?

AIMS: Cystic fibrohistiocytic tumour of the lung is a rare proliferative process. Its histogenesis is uncertain, but evidence suggests that some cases represent metastatic disease from apparently indolent skin lesions, namely cellular fibrous histiocytomas. This study presents four cases and reviews the literature concerning this pattern of disease and its aetiology. METHODS AND RESULTS: All patients were male (age range 35-54 years). Two presented with recurrent haemoptysis. Two cases had histories of cutaneous fibrohistiocytic lesions in the chest wall, excised 10 and 23 years prior to presentation with lung disease. Imaging data showed multiple bilateral cystic lung lesions in all four patients with nodular cavitating opacities seen on high-resolution computed tomography scans. Microscopy showed variably dilated thin-walled cystic airspaces lined by cuboidal epithelium and an underlying layer of mildly pleomorphic spindle cells with slightly wavy morphology and storiform architecture, admixed with inflammatory cells. Tumour cells stained for CD68 in three of four cases. All cases were negative for CD34. All patients were alive with disease, although one required pneumonectomy for intractable haemoptysis. CONCLUSION: This study and a review of published cases show that the majority of cystic fibrohistiocytic tumours of the lung probably represent metastases from cellular fibrous histiocytomas. However, rare cases may be either primary in origin or the primary site remains occult; the term cystic fibrohistiocytic tumour remains appropriate for such cases.

[Carcinoids of the gastrointestinal tract: importance of determining differentiation and proliferation markers]. / Karcinoidy gastrointestinálního traktu: význam hodnocení diferenciacních a proliferacních markeru.

The group of 35 carcinoid tumours obtained from 34 patients was reviewed according to recent histopathological criteria. Consequently, evaluation of the Grimelius staining and immunohistochemical detection of chromogranin A (CgA), Leu-7 (CD-57), synaptophysin, neuron-specific enolase (NSE), (beta-III tubulin, Ki-67 and proliferating cell nuclear antigen (PCNA) was performed. The majority of tumours (29, i.e. 83%) were classified as typical carcinoids composed predominantly of mixed solid and trabecular or solid and tubular growth patterns. Six tumours (17%) revealed more prominent cytological abnormalities corresponding with the diagnosis of atypical carcinoid. The majority of tumours (31, i.e. 93.9%) showed granular cytoplasmic positivity in Grimelius staining and diffuse cytoplasmic positivity of NSE (34, i.e. 97.1%). All of the 32 stained tumour samples showed positive immunoreactivity for synaptophysin. A high percentage of tumours (32, i.e. 91.4%) revealed also a positive reaction with antibody TU-20 detecting (beta-III tubulin, a marker of an early stage of neuronal differentiation. Thirty-four tumours (97.1%) showed granular cytoplasmic positivity for both markers of neuroendocrine granules (CgA and Leu-7). One tumour (2.9%) was positive only for Leu-7. Tumour cells revealed predominantly low proliferative activity evaluated by PCNA and Ki-67 immunodetection. Higher degree of proliferation was observed especially in atypical carcinoids.

Different patterns of chromogranin A and Leu-7 (CD57) expression in gastrointestinal carcinoids: immunohistochemical and confocal laser scanning microscopy study.

Thirty-seven carcinoids of the gastrointestinal tract were studied with immunohistochemical staining for chromogranin A (CgA) and Leu-7 (CD57). The aim of this study was to distinguish and describe the differences in patterns of distribution of immunostaining of these two non-specific neuroendocrine markers in neuroendocrine tumors of different degree of differentiation (typical, vs. atypical carcinoids) at different gastrointestinal sites. Selected 5 tumors from this group were studied in detail using confocal laser scanning microscopy (CLSM) and double immunofluorescence staining to disclose the patterns of distribution of CgA and CD57 positive granules within the individual tumor cells. Prominent differences in the patterns of immunohistochemical staining for both studied markers related to the degree of differentiation of the tumors were observed in studied neoplasms. Regular (diffuse) strongly positive immunoreaction for CgA predominated in typical carcinoids, whereas atypical tumors were characterized by irregular patchy staining. Both typical and atypical tumors displayed predominantly irregular patchy staining for CD57. The results of CLSM study indicate that different modes of CgA and CD57 expression and/or co-expression can occur in neuroendocrine tumors. Neoplastic cells that contained either CgA positive neuroendocrine granules (NEG), or Leu-7 positive NEG, were frequently observed in different areas of the tumor samples, especially in atypical carcinoids. Varying number of cells revealed co-localisation of both CgA and Leu-7 within the NEG. Similar co-localisation of CgA and CD57 was found in non-neoplastic Kultschitski cells of the mucosa of small intestine. In conclusion, our results suggest that the differences in CgA and CD57 expression in human neuroendocrine tumors are related to the degree of differentiation of the neoplasms and probably reflect the degree of maturation (functional state) of neuroendocrine granules within the neoplastic cells.

Expression of class III beta-tubulin in neuroendocrine tumours of gastrointestinal tract.

Class III b-tubulin is presented as a specific marker for the cells of neuronal origin as well as for the tumours originating from these cells. Its expression is considered one of the earliest events that appear in the cells revealing neuronal differentiation. Using monoclonal antibody TU-20 in an immunohistochemical analysis, we studied the expression of class III b-tubulin in gastrointestinal carcinoid tumours. Paraffin-embedded, formalin-fixed tissue sections from 49 tumour samples obtained from following locations: stomach (4 cases), small intestine (8 cases), appendix (18 cases), rectum (3 cases), pancreas (5 cases), liver metastases (7 cases) and lymph node metastases (4 cases) were used in the study. In 41 of the 49 tumour samples (83.7%), positive staining for class III b-tubulin was detected, while 8 tumour samples (16.3%) were negative. Expression of class III b-tubulin was prominent in all three rectal carcinoids and in three atypical carcinoids located in small intestine. Pancreatic neuroendocrine tumours revealed either weak immunostaining (2 cases), or were negative for this marker (3 cases). The intensity of class III b-tubulin immunolabelling was not related to the degree of tumour differentiation. The results of this study suggest that class III b-tubulin could be a perspective marker for gastrointestinal neuroendocrine tumours. Moreover, the differences in its expression could also elucidate some aspects of histogenetic relationships of neuroendocrine tumours of gastrointestinal tract.

Relevant animal model of human lymphoblastic leukaemia/lymphoma--spontaneous T-cell lymphomas in an inbred Sprague-Dawley rat strain (SD/Cub).

More than a decade of experimental work in an inbred subline of Sprague-Dawley rats having high incidence of spontaneous T-cell lymphoma/leukaemia is reviewed. Longitudinal follow-up of biological characteristics (growth, survival, haematology) of both multiple cases of primary disease and s.c. passaged lymphomas as well as comparative immunophenotypic and karyotypic studies are concluded. In these T-cell lymphomas (mostly CD4 positive), arising on the same genetic background of the inbred SD strain, the aberrations involving chromosome 11 have been recognized as a typical non-random cytogenetic marker. This unique rat model of lymphoblastic lymphomas/leukaemias, relevant to human pathology, seems to be very suitable for testing different anticancer therapeutic strategies, as it is documented by results of a number of various protocols conducted in our laboratory.

Embryotoxicity of cisplatin and a cisplatin-procaine complex (DPR) studied in chick embryo.

Cisplatin is widely used as an antitumor drug. To reduce its toxic side effects in patients, cisplatin has been bound with procaine in a cisplatin-procaine complex (DPR). The lethal and teratogenic effects of cisplatin alone and of complexed cisplatin were determined in the chick embryo in ovo in order to compare their influence on rapidly proliferating embryonic tissues. The embryotoxic (lethal + teratogenic) effect was examined after a single intra-amniotic injection of one of six different doses, ranging from 0.03 to 30.0 microg, on embryonic days (ED) 3, 4 or 5. The minimal embryotoxic dose was lower for cisplatin alone (0.03-0.3 microg) than for cisplatin in the DPR complex (0.3-3.0 microg), suggesting that cisplatin alone is more embryotoxic than complexed cisplatin. Both substances caused malformations in the surviving embryos evaluated on ED 9. These malformations included microphthalmia, microcephaly, hypoplasia of the upper and lower jaw, cleft beak, and haemocephaly. Moreover, heart septum defects and limb reduction deformities were found after exposure to the DPR complex. The embryotoxicity of complexed cisplatin exhibited a stage-response effect. It was highest on day 3 and gradually decreased until ED 5. Such an apparent stage-response effect was not observed for cisplatin alone. The embryotoxicity of procaine hydrochloride - a component of the complex - was also tested. Procaine hydrochloride alone did not produce any embryotoxic effect, not even after a single injection of the maximal tested dose (100.0 microg per embryo). We also examined the protective effect of procaine hydrochloride, whose separate administration at ED 4 was followed by the injection of 0.3 microg cisplatin. We did not observe any protective effect of procaine hydrochloride if injected separately.

Antitumour activity of a combined treatment with PMEDAP and docetaxel in the Prague inbred Sprague-Dawley/cub rat strain bearing T-cell lymphoma.

Antitumour efficiency of combined therapy with N-9-[2-(phosphonomethoxy)ethyl]-2,6-diaminopurine (PMEDAP) and docetaxel (DTX) was studied in an in vivo model of s.c. transplanted Sprague-Dawley (SD/Cub) rat T-cell lymphoma (phenotype SD10/96). The effect of the combined treatment of DTX with PMEDAP was significantly higher than that of DTX or PMEDAP alone. The s.c. administration of DXT into the vicinity of growing lymphoma together with i.p. administration of PMEDAP was found to be the most efficient combination. In this case, two out of four rats did not develop any lymphoma and remained alive. An irregular expression of Bcl2 protein was found in untreated and treated lymphomas, while the expression of protein p53 as well as MDM2 was not observed. All three types of the above-mentioned treatments (PMEDAP, DXT, DXT+PMEDAP) increased significantly the number of p21-positive cells, compared with untreated tumours.

[Epithelioid hemangioendothelioma of the liver (case report)]. / Epiteloidní hemangioendoteliom jater (kazuistika).

Epitheliolid hemangioendothelioma of the liver is a very rare vascular tumor. We report the case of this tumor found during the autopsy of a 74-year-old man. The liver was infiltrated by multiple greyish nodes (diam. 3-30 mm), partially calcified. Microscopically, the tumor was composed of dense sclerotic and fibrous stroma with focal calcifications. In this stroma, groups and cords of irregular cells of two main types were dispersed. The first type was represented by big cells with large clear eosinophilic cytoplasm. The second cell type was characterised by medium-sized elongated, sometimes bizarre-shaped cells. Tumorous cells did not produce mucin. Immunohistochemically, the tumorous cells were positive for FVIIIR--Ag, actin, vimentin, and desmin and also showed disperse weak positivity for S100 protein. Staining for epithelial membrane antigene (EMA), keratins, and CD68 was negative. Neoplastic cells were positive for Ulex europaeus lectin. Electronmicroscopically, the tumorous cells had irregular nuclei, lobular or cleft shape with one or two nucleoli. Cytoplasm contained a high amount of intermediate filaments forming large aggregates. The cells were connected by desmosomes and partially surrounded by dense basal lamina. The diagnosis of epithelioid hemangioendothelioma is based on a histological picture, supported by immunohistochemical positivity for FVIIIR: Ag and Ulex europaeus and by additional ultrastructural study.