The authors described the localization of neuron-specific enolase in the intestinal wall in Crohn´s disease. We have used samples obtained by biopsy from the colon lining of five people affected by Crohn's disease for our examination. We have processed samples using the formol paraffin technique. From paraffin blocks, we have prepared histological sections approximately 5 μm thick. For immunohistochemic examinations, we have revitalised the sections by acquiring the heat-induced epitope. We detected NSE by monoclonal mouse antibodies against human neuron-specific enolase, clone BBS/NC/VI-H14 (DakoCytomation, Denmark) (Fig. 4, Ref. 7).

Breast carcinoma is the most common cancer with high mortality caused by metastatic disease. New molecular biomarkers predicting the tumour's metastatic potential would therefore improve metastasis prevention and personalised care. The aim of the study was to investigate the relationship between DNA methylation levels in invasivity and metastasising associated genes with aberrant protein expression and also to evaluate whether a similar DNA methylation level is present in the tumour and circulating cell-free DNA for utilising plasma DNA methylation as prognostic biomarker. By using pyrosequencing, we analysed DNA methylation levels of 11 genes, namely APC, ADAM23, CXCL12, ESR1, PGR B, CDH1, RASSF1A, SYK, TIMP3, BRMS1 and SOCS1 in tumour, plasma and peripheral blood cells from 34 patients with primary breast cancer, as well as plasma and peripheral blood cells from 50 healthy controls. Simultaneously, the expression of related proteins in paraffin-embedded tumour samples was evaluated by immunohistochemistry. Statistical analysis was performed by SPSS statistics 15.0 software. Tumour DNA hypermethylation was found in most commonly methylated RASSF1A (71.9%), APC (55.9%), ADAM23 (38%) and CXCL12 (34.4%) genes with methylation levels up to 86, 86, 53 and 64 %, respectively. In tumours, significantly higher methylation levels were found in nine genes, compared with the patients´ peripheral blood cell DNA. Furthermore, in patients methylation levels in peripheral blood cell DNA were significantly higher than in controls in CXCL12, ESR1 and TIMP3 genes, but the values did not exceed 15%. On the other hand, no correlations were observed in patients between DNA methylation in tumours and cell-free plasma DNA. Moreover, in patients and controls nearly identical values of cumulative DNA methylation (43.6 % ± 20.1 vs. 43.7 % ± 15.0) were observed in plasma samples. A variable spectrum from high to none expressions presented in tumour tissues in all of the proteins evaluated, however in APC and CXCL12 genes a visible decreasing trend of mean DNA methylation level with increasing expression of the corresponding protein was observed. The DNA methylation profiles manifested in our group of breast carcinomas are cancer specific, but they are not the only cause that affects the silencing of evaluated genes and the decrease of relevant protein products. The clinical utility of DNA methylation testing in peripheral blood cell DNA for cancer diagnosis and therapy need to be further investigated.

AIM OF THE WORK To describe the state of microcirculation in the intestinal wall in ulcerative colitis and Crohn's disease in own material. MATERIALS AND METHODS For morphological examination, we obtained tissue samples from both diseases during bioptic bowel examination from 11 patients. We fixed the samples with 10% buffered formalin, de-hydrated and covered with paraffin. From paraffin blocks, we made histological sections about 5μm thick with a microtome. They were always stained with haematoxylin and eosin. For immunohistochemistry sections, we either did or did not revitalise the sections according to the manufacturer's recommendations for specific to-reagents. For immunohistochemic examinations, we revitalised the sections by acquiring the heat-induced epitope in DakoCytomation Target Retrieval Solution, Code No S 1700, or in mmol.l-1 citrate buffer, pH 6.0. We did not dry the sections during such procedure. We used antibodies as follows: Monoclonal mouse antibodies against human von Willebrand factor, Clone F8/86 (DakoCytomation, Denmark), Monoclonal mouse antibodies against smooth muscle actine, Clone HHF35 (DakoCytomation, Denmark). We used detection system Dako EnVisionTM + Dual Link System-HPR (Dako, Denmark) or Dako Liquid DAB + Substrate Chromogen System (Dako, Denmark) to display areas of specific connection of antibodies. Antibodies against von Willebrand factor react in our samples with the endothelium of vessels and with precursors in the lining of peaks of villi. We have also seen a diffuse positive reaction. Lymphatic vessels do not display monoclonal antibodies (Fig. 4, Ref. 8).

INTRODUCTION:The most risk group with high rate of infection of viral hepatitis C represents the former drug abusers (IDUs).OBJECTIVES:The objectives of the clinical study were to assess the sustained virological response (SVR) in the group of former IDUs with chronic hepatitis C (CHC) treated with pegylated interferon-α and ribavirin.PATIENTS AND METHODS:Of 293 previously untreated (naive) former IDUs with CHC who have started treatment, we assessed 239 (69 women, 170 men) with an average age of 27 years who completed treatment and received a follow-up examination after 24 weeks. Prior to therapy abstinence to drug abuse 6 and more months has been required. The CHC has been diagnostics by the standard virological diagnostic tests, increased activity of aminotranferases (ALT) and histology of liver biopsy. Patients were treated with combined immunomodulatory therapy in the standard duration and standard regimen. SVR was considered if qualitative test HCV RNA was negative 24 weeks after the completion of the treatment.RESULTS:End-of-treatment virological response was in 224/94 % patients. Overall SVR rate was 95 %, 227 subjects had negative HCV RNA and only 12 patients no response. During treatment there was a statistically significant decrease in ALT. Treatment was accompanied by the expected side effects.CONCLUSION:Young age, short duration of HCV infection, high prevalence of genotype 3 and low grade of liver fibrosis in majority of patients and good adherence of patients to treatment were crucial predictive factors resulting in high SVR.

WHO Classification of Tumours of the Central Nervous System from the 2007 is distinguished from the previous 2000 classification by a few conceptual modifications, changes in the terminology and seven newly codified tumour entities. The text shows a short comparison of both classifications emphasising the most important changes from the surgical neuropathology point of view. The newly codified entities are: angiocentric glioma, pilomyxoid astrocytoma, papillary glioneuronal tumor, rosette-forming glioneuronal tumor of the 4th ventricle, papillary tumour of the pineal region, spindle cell oncocytoma and pituicytoma. Mostly, they are rare tumours already known from the literature. Based on new knowledge from the molecular pathology the paragraphs about tumour genetics were markedly changed. The complexity and diversity of tumours of the nervous system is enormous, and, not surprisingly, some problematic questions of classification and grading remain unresolved.

Glomerulopathies with fibrillary deposits form a heterogeneous group of renal diseases that can be identified only by means of electron microscopy. A case of a rare type of such a nephropathy, the collagenofibrotic glomerulopathy with focus on differential diagnostics is presented and current knowledge relating to this renal disease is reviewed.

Five cases of epidermoids in suprasellar region, which underwent surgery from 1996 until 2000, were retrospectively reviewed (four females, one male, and hyperintense. After injection of contrast medium in all five cases was enhancement on periphery (from mild to thick). At surgery except of usual yellowish debrits, there were present areas of inflammatory tissue and brown viscoid material. On histology keratin, cholesterol crystals, cells and tissue of chronic inflammation, acute or chronic haemorrhage were present (table 1).

OUTLINE The paper presents the results of 33 microsurgical removal of 33 tumours of brachial plexus and of peripheral nerves in extremities performed on 31 patients during the period of 11 years, from 1990 to 2001. PATIENT GROUP AND METHODS The patient group included 26 patients treated for removal of 28 tumours of peripheral nerves in extremities and 5 patients treated for removal of 5 tumours of brachial plexus. Three of these brachial plexus tumours showed intraspinal spread. 32 tumours of neural sheath included 20 schwannomas and 12 neurofibromas. 6 of these 12 neurofibromas were observed in 4 patients suffering from Reklinghausen disease (VRD). One patient had a tumour of non-neural elements--lipoma. We did not encounter any malign tumours of neural sheaths or other neural sheath-derived tumours (benign or metastatic malign ones) that would compress or invade the nerves or plexus. The results were evaluated according to Donner classification of motor and sensory functions. RESULTS Motor functions improved or remained unchanged in 18 (90%) patients with schwannoma. 5 patients (83.3%) with pain in the distribution of the affected nerve reported complete or partial improvement of the symptoms. Regarding neurofibroma patients, 3 exhibited either an improvement or unchanged motor functions, and all of them reported partial or complete retreat of pain symptoms. 4 patients with VRD were subjected to the removal of 6 tumours of peripheral nerves in extremities; all of them exhibited improved or stabilised motor functions and partial or complete retreat of pain symptoms. The outcome of treatments was generally better in smaller tumours of neural sheaths with more distal localisation. CONCLUSION Patients should be treated in the early stage of tumour development when the size remains smaller and neurological deficits are absent.

AIM OF THE STUDY:Investigation of the relationships between the grade and stage of chronic liver diseases irrespective of their etiology using some novel serum markers of liver fibrogenesis, the "classical" serum markers of liver necro-inflammatory injury (such as transaminases), and the histomorphological evaluation of liver biopsies.METHODS:Markers of liver fibrogenesis: serum metalloproteinase 1 (MMP-1), tissue inhibitor of MMP-1 (TIMP-1), and N-terminal propeptide of the procollagen III (PIIINP); "liver function tests" (LFTs): bilirubin, transaminases ALT, AST; ALP, GMT; and liver morphology findings: necro-inflammatory activity, fibrosis; were studied in the series of 32, 'naive', i.e. yet untreated patients (women/men--11/21) with various CLDs: chronic viral hepatitis B or C 13 (CHB 3, CHC 10), non-alcoholic steatohepatitis 9, liver steatosis 4, primary biliary cirrhosis 5, drug-induced hepatitis. The diagnoses were based on the clinical, laboratory and liver imaging (ultrasonography) findings and confirmed by the liver biopsy.CONCLUSIONS:Investigation of liver fibrogenesis serum markers (PIIINP, MMP-1, TIMP-1) in patients with various CLDs has shown statistically significant correlations of these parameters with "classical" serum markers of liver necro-inflammation (ALT, AST) and the results of histomorphological evaluation of the necro-inflammatory activity (parameters NAI, MEF) and fibrosis (parameter FI) in liver biopsies. (Tab. 4, Ref. 31.)

OBJECTIVE:In organ allotransplantation represents a serious problem acute rejection, which is the reaction of recipient organism aimed to reject the transplanted organ.AIM:To obtain experience in the field of routine long term are of patients after heart transplantation living in Slovakia.METHOD:33 patients after orthotopic heart transplantation (HTx) were included in our long term care by the 1. June 1996. According to the accepted protocol we make the whole set of examinations is performed. This includes endomyocardial biopsy which is the only reliable method for the detection of rejection and also the criterion of sufficiency of immunosuppressive therapy.RESULTS:Acute rejection was revealed in 14 (43%) patients.CONCLUSIONS:Tactics of therapy and the length of the interval to subsequent biopsy were determined according to the histological findings of preceding biopsy, the clinical status and the combination of immunosuppression therapy. (Tab. 2, Ref. 13.)

The authors present an account on contemporary knowledge of the diagnosis and differential diagnosis of fibrillar glomerulopathies. The latter are characterized by extracellular localized microfibrils and microtubules resp. in the glomeruli of the kidneys, their diameter being 8-60 nm. They are divided into amyloid and non-amyloid types. The others are classified according to the immunofluorescent finding into immunoglobulin positive and negative ones. The differential diagnosis is important in particular in immunoglobulin positive ones as they are present in serious diseases such as cryoglobulinaemia, monoclonal gammapathy, systemic lupus erythematosus and immunotactoid glomerulopathy.

Authors described their experiences with angiocardiography, (selective coronarography and left ventriculography), in 41 patients aged from 32 to 59 years with the preliminary diagnosis of cardiopathy of unknown origin. The patients with diagnosis of cardiopathy of unknown origin were those, who cannot be assigned to any of the known and acknowledge diagnostic categories, not even by using complex clinical, laboratory and noninvasive cardiological diagnostic procedures. Angiocardiography enabled to produce new and unexpected data of diagnostic value three patients with heart disease (all of the three mentioned patients had significant changes of the coronary supply, i.e. the reduction in lumen diameter of the coronary vessel being of more than 50% with local or total wall motion abnormalities). The other thirty-eight patients had normal findings of the coronary supply. One patient had a normal coronarographic findings, normal left ventricular wall motion and also the subsequent histological examination from the endomyocardial bioptic sample of the heart tissue, did not confirm pathological abnormalities. Selective coronarography and left ventriculography is a useful and safe clinical diagnostic procedure in exactly defined groups of patients.

Three-sialic acid-specific lectins, Tritrichomonas mobilensis (TML) (without linkage preference), Maackia amurensis leukoagglutinin (MAL) (alpha 2, 3) and Sambucus nigra agglutinin (alpha 2,6 linkage-specific), were used for detection of sialylated glycoconjugates in normal and diseased human kidneys. Normal kidneys demonstrated strong podocyte positivity of alpha 2,3 linked sialic acid and weaker sialic acid expression on capillary endothelium, which was alpha 2,6 linked. Renal biopsies (45) representing a variety of diseases, e.g. minimal change disease, membranous, membranoproliferative glomerulonephritis, tubulointerstitial nephrosis or diabetic glomerulopathy showed increased sialic acid expression in glomerular capillaries, Bowman's capsule epithelium and on podocytes. In several different kidney diseases the glomerular endothelium expressed also alpha 2,3 linked sialic acid along with increased TML-positivity of epithelial cells related to alpha 2,6 linked sialic acid. No-difference in sialic acid linkage expression was observed in the tubules, which expressed putative alpha 2,6 linked sialic acid on the luminal surface of cells distal to the descending limb of Henle's loop. The study did not show changes characteristic of specific diseases. Rather, increased sialic acid expression on glomerular endothelium and podocytes was linked to a variety of pathological changes. It is assumed that changes in sialysation of glycoconjugates in the glomeruli represent nonspecific changes and do not reflect fundamental pathogenetic features of renal diseases.

The authors described aim. objectives, design and criteria of the study which is oriented to the clinical and research problems of heart diseases of uncertain origin. The paper presents the preliminary experience with invasive cardiological techniques in this field. At the same time the authors discuss the potential impact of the basic research on the prevention and effective treatment of cardiopathy which is until now of unknown origin.

Authors measured the thickness of the basement membranes of the proximal and distal tubuli in the renal cortex in 20 patients with verified diabetes mellitus. The test group consisted of 20 patients with small abnormalities of glomeruli (or in the range of variable norm), of the same sex and comparable age. Measurements were performed in locations without tubulo-interstitial changes. The average thickness of TBM in patients with diabetes mellitus was 1540±616 nm, and in the test group 360±86 nm, which is a statistically significant difference (p<0.000000). In spite of the small number of cases and possible error in the manual measurement of electron microscopic pictures, a significant thickening of TBM in diabetes mellitus has been found. We consider this to be of diagnostic value.

The authors describe four, out of 506 cases of renal biopsies in which this diagnostic procedure enabled to reveal a tumour. Two cases of tumours which were intentionally subdued to renal biopsy justified an angiomyolipoma and a focus (metastasis) of tubulopapillomatous carcinoma) in the kidney. In one case the renal biopsy revealed renal infiltration thus enabling to state the diagnosis of chronic lymphatic leukaemia subsequently, and in the last case where the diagnosis of chronic lymphatic leukaemia had been stated in advance, the renal biopsy was performed due to suspective glomerulonephritis. In these cases the unexpected results of renal biopsy were of great diagnostic value. Such intentional use of biopsy is rare. (Fig. 3, Ref. 6.)