[Can Analysis of Cellular Lipidome Contribute to Discrimination of Tumour and Non-tumour Colon Cells?] / Muze analýza bunecného lipidomu prispet k rozlisení nádorových a nenádorových bunek tlustého streva?

BACKGROUNDS: Colon cancer development is often characterized by abnormalities in lipid synthesis and metabolism, which may influence energetic balance, structure and function of biological membranes, or production of specific mediators and cell signalling. The changes in lipid profile and metabolism (lipidome) may significantly affect cell behaviour and response to therapy. Permanent epithelial cell lines at various stages of cancer development are used for better understanding of this topic on cellular and molecular levels. In our study, we hypothesized that detailed analyses of colon cancer cell line lipidomes may help to identify major alterations in the amount and profile of specific lipid classes/species, which can contribute to their different response to various stimuli. MATERIAL AND METHODS: Cellular lipids were isolated from six human epithelial cell lines derived from tissues at various stages of tumour development. Liquid chromatography coupled with tandem mass spectometry analyses were performed in order to determine amount and mass profiles of all phospholipid (PL), lysophospholipid (lysoPL) and sphingolipid classes. The data was statistically evaluated (cluster and discrimination analyses) with respect to mutual comparison of cell lines and to significantly discriminating lipid types. RESULTS: The results of cluster analysis arranged cell lines in order corresponding to their level of transformation (normal cells, adenoma, carcinoma and lymph node metastasis). The results of discrimination analyses revealed the most discriminating lipid types and distinction in PL: lysoPL ratios. Particularly, significant correlation of the amount and profiles of both specific lysoPL and sphingolipid classes with cell transformation level were observed. Similar approaches are now applied to compare lipidomes of colon epithelial cells isolated from tumour vs. non-tumour samples of colon cancer patients. CONCLUSION: Our results indicate that a) selected cancer cell lines are suitable model for lipidomic studies that can serve as a basis for subsequent clinical research, b) cellular lipidome analyses may help to discriminate tumour and non-tumour cells in clinical samples, where specific types of lipids could serve as biomarkers.Key words: colon cancer - cell lines - liquid chromatography - mass spektrometry - phospholipids - sphingolipids - bioinformatics The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. This work was supported by Czech Health Research Council, grant No. AZV 15-30585A.Submitted: 19. 3. 2018Accepted: 18. 4. 2018.

Non-Small Cell Lung Cancer with Estrogen Receptors and ALK Positivity.

BACKGROUND: Estrogens are steroid hormones that affect a number of physiological functions in cells; these compounds play an irreplaceable role in the reproductive system. Their effects are mediated by the estrogen receptor (ER), of which there are two subtypes - ERα and ERβ. ERs are expressed in regions outside the reproductive system, including bone, brain, intestine, endothelium, kidney, and lung. Expression of ER by lung cancer (LC) cells was first described in the early 1980s. The experimental literature also describes co-expression of ERβ and an epidermal growth factor activating mutation, and the additive antiproliferative effects of anti-estrogens plus tyrosine kinase inhibitors during treatment of lung adenocarcinoma. However, coincident expression of ER and ALK translocation were not decribed. The 4-year survival for generalized non-small cell lung cancer indicates the possibility that both molecular features (ER and ALK positivity) may be a favorable prognostic biomarker for this tumor. CASE: A 69-year-old patient presented with generalized lung adenocarcinoma that was positive for ERb and the ALK-EML4 fusion gene. The tumor was stage IV. Additional examinations excluded malignancy of the gynecological area. The patient was treated with chemotherapy and a tyrosine kinase ALK inhibitor. CONCLUSION: The role of individual ER subtypes in LC carcinogenesis, and the possible therapeutic effects, is unclear. This is the first documented case of co-occurrence of ALK-EML4 and ERβ in LC. Key words non-small cell lung cancer - estrogen receptors - ALK translocation - prognosis - treatment This work was supported by Ministry of Health of the Czech Republic, grant AZV 16-32318A, and by Ministry of Education, Youth and Sports of the Czech Republic, grant NPU I LO1304. The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 23. 9. 2018 Accepted: 25. 10. 2018.

[Validity of the Morawietz classification for evaluation of periprosthetic tissue]. / Prínos Morawietzovy klasifikace k hodnocení periprotetických tkání.

PURPOSE OF THE STUDY A consensual classification of the periprosthetic interface membrane obtained at revision total joint arthroplasty was published by Morawietz et al. in 2006. Based on histomorphological criteria, four types of periprosthetic membrane were proposed: type I, aseptic failure; type II, septic failure; type III, combined type (carrying signs of both type I and II); and type IV, indeterminate type. The aim of this study was to find out whether and to what extent the Morawietz system would be suitable for use at an independent institution involved in the evaluation of periprosthetic membranes for a long time. Should it appear that the institution achieved an equally good or even better agreement between the clinical diagnosis and the histopathological finding, this consensus classification could be recommended for routine use. MATERIAL AND METHODS The samples of periprosthetic tissue evaluated in this study were obtained during surgery from the following groups of patients: 66 patients with aseptic loosening of total hip (THA) or knee arthroplasty, 15 patients with infection of THA, 16 patients with THA without any signs of aseptic loosening, osteolysis or infection; 8 patients with hip osteoarthritis and 8 patients with knee osteoarthritis. Sample collection and processing (for purposes of histomorphological evaluation and immunohistochemical staining) was performed according to the established protocol. The tissue samples evaluation was made by an experienced pathologist hand in hand with the method described in the original paper by Morawietz et al. For a more detailed tissue analysis, selected antibodies (CD4, CD8, CD20, IFN-γ and Hsp-60) were visualized by immunohistochemistry. RESULTS The majority of samples from aseptic reoperations were classified as membranes of the type I (79%) and III (16%). Specimens retrieved from septic cases were mostly classified as membranes of type II and III (60% together). The septic membranes showed a significantly higher expression of CD20 protein when compared with both the aseptic (p < 0.0001) and control THA samples (p = 0.003). The membranes retrieved from the surroundings of a stable THA without osteolysis and infection had lower expression levels of Hsp60 and IFN-γ, when compared with those from both aseptic and septic loosening. Finally, Hsp-60 expression was significantly higher in osteoarthritic tissue than in samples from stable THA (p = 0.041). DISCUSSION Morawietz et al. proposed a standardized classification system for evaluation of periprosthetic tissue. As any attempt at generalization of a complex issue, this proposal has certain shortcomings. One of these is poor detection of chronic and low-grade infections. A method that would improve the conventional counting of polymorphonuclear leukocytes is still being sought. In this connection, immunostaining for CD20 combined with an assessment of antimicrobial peptides may be a promising option. The supplementary specimen staining showed that pseudosynovial tissue is much more active in patients carrying infection and the least active in samples from stable THA in which certain tolerance and thus tissue homeostasis might be expected. CONCLUSIONS 1. In this study the distribution of findings classified according to the Morawietz system was similar to the results published in the original study from 2006. 2. The definition of an aseptic membrane (type I) in the Morawietz system meets the requirements of clinical practice (agreement, about 80%). 3. An increased sensitivity for infectious membrane detection can be achieved by using supplementary immunohistochemical staining effective particularly in chronic and low-grade infections. 4. Painless and stable THAs typically have very low expression levels of CD4, CD20 and Hsp-60 proteins, and interferon- -gamma (IFN-γ) as well. Key words: total hip arthroplasty, total knee arthroplasty, aseptic loosening, prosthetic joint infection, tissue analysis, membranes, CD receptors, Hsp-60 protein, IFN-γ.

[Low expression of Tlr-1, -2, -4 a il-2, -2r, -10, -10r and TgFb1 in pseudosynovial tissue collected at revision total knee arthroplasty for aseptic loosening]. / Nízká exprese Tlr-1, -2, -4 a il-2, -2r, -10, -10r, TgFb1 v pseudosynoviální tkáni odebrané pri reoperaci asepticky uvolnené totální endoprotézy kycle a kolena.

PURPOSE OF THE STUDY: Aseptic loosening (AL) and periprosthetic osteolysis (PPOL) in total hip (THA) and knee (TKA) arthroplasty are linked to an inflammatory process initiated by wear debris released from artificial joints. There is still limited information about the contribution of Toll-like receptors (TLRs) and distinct regulatory cytokines to AL/PPOL in both joints. METHODS: In this study, we investigated mRNA expression of TLR-1,-2,-4 and cytokines/receptors (IL-2,-2R,-10,-10R, TGFb1) in pseudosynovial tissue obtained from 55 patients with aseptically failed THAs/TKAs and 37 control patients with hip/knee primary osteoarthritis (OA) using quantitative RT-PCR. Immunohistochemical staining was used to detect the corresponding proteins. Non-parametric Kruskal-Wallis and Mann-Whitney tests were used to determine differences between the patient groups. RESULTS: When comparing expression profiles between patients with aseptically failed THA and TKA, higher amounts of TLR-1,- 2,-4 and IL-2R mRNA transcripts were detected in THA patients. The mRNA expression of studied molecules (TLR-1,-2,-4, IL-2, IL-10, IL-2R, IL-10R, TGFb1) did not differ between THA and OA hip tissues. Lower mRNA expression of TLR-1,-2,- 4, IL-10, and IL-10R was detected in TKA when compared to control knee OA. Similar mRNA profiles of IL-2, IL-2R, and TGFb1 were observed in TKA and knee OA. Using immunohistochemistry, we detected low expression of TLR-1 protein in failed THA/TKA, whereas TLR-2 protein levels were higher in TKA/THA patients than in OA controls. High individual variability in TLR-4 protein levels was detected among patients with aseptically loosened THA and TKA. IL-10 protein levels were similar in THA and TKA patient subgroups and control subjects, whereas IL-10R protein level was higher in failed TKAs and OA controls than in THAs. No difference in IL-2 protein levels was detected between patients with THA/TKA and those with OA. DISCUSSION: Our data indicate close similarity between the expression patterns in aseptically failed THA and TKA. However, certain differences were observed which also suggest unique pathways associated with the end-stage of aseptic loosening in THA and TKA. For instance, differences in the size, shape and load of polyethylene particles between THA and TKA could play some role. The composition of THA and TKA and differences in terms of mechanical forces might also be involved. CONCLUSIONS: This is the fist study comparing the gene expression profile of a particular set of innate immunity regulatory molecules between tissues from aseptically failed THA and TKA. Low expression of TLR-1,-2,-4 and cytokines/receptors (IL-2, IL-2R, IL-10, IL-10R, and TGFb1) was observed in pseudosynovial tissues obtained from aseptically failed THAs and TKAs. Higher amount of TLR transcripts was detected in THA as compared to TKA. These findings indicate certain differences in the mechanism of aseptic loosening occurring at the site of THA and TKA. Further research is warranted.

[Surgical therapy of pancreatic cancer - 5 years survival]. / Resekabilní karcinom pankreatu 5leté prezití.

INTRODUCTION: The purpose was to identify 5-year survivors among a group of radically resected patients with pancreatic cancer and analyse the characteristics and factors associated with their 5-year survival. Single tertiary centre experience. METHOD: A prospectively maintained database of 155 pancreatic resections from January 2006 to June 2010 was scanned to identify patients after curative radical resections for pancreatic ductal adenocarcinoma. The clinical and pathological data was analysed retrospectively. The outcomes of the PDAC group were evaluated using Kaplan-Meier analysis (survival) with the Log-rank test and Cox regression analysis (evaluation of prognostic factors). Characteristics of the survivors were discussed. Significance level of 0.05 was used. Those factors were used as independent variables for Cox regression analysis whose significant effect on survival was shown based on Kaplan-Meier analysis. RESULTS: Among 155 patients undergoing a curative pancreatic resection, 73 had a pancreatic ductal adenocarcinoma. Fifteen patients (20.5%) after radical surgery survived over 5 years, 13 of whom are still alive. In the group of the survivors, the mean overall survival was 77.1 months (60110) and the median survival was 74 months. The mean relapse-free interval in the group of the survivors was 63.3 months (14110) with the median of 65 months. Factors associated with a longer survival included the absence of lymph node infiltration (p=0.031), uncomplicated postoperative course (p=0.025), absence of vascular invasion (p=0.017), no blood transfusions (p=0.015) and the use of postoperative therapy - predominantly chemotherapy (p=0.009). Significant independent predictors of survival included vascular invasion HR=2.239 (95%CI: 1.0934.590; p=0.028), postoperative chemotherapy HR=2.587 (95%CI: 1.3015.145; p=0.007) and blood transfusion HR=2.080 (95%CI: 1.0274.212; p=0.042). The risk of death was increased 2.2 times in patients with vascular invasion, 2.1 times in patients with transfusions, and finally 2.6 times in those with no chemotherapy. CONCLUSION: Factors associated with an improved overall survival included: the absence of lymph node infiltration, an uncomplicated postoperative course, absence of vascular invasion, no need of blood transfusions, and finally the use of postoperative chemotherapy. Vascular invasion, use of blood transfusions and postoperative adjuvant chemotherapy were significant independent prognostic factors of survival.

Comparison of periprosthetic tissues in knee and hip joints: differential expression of CCL3 and DC-STAMP in total knee and hip arthroplasty and similar cytokine profiles in primary knee and hip osteoarthritis.

OBJECTIVE: To identify expression profiles (EP) associated with aseptic loosening of total knee arthroplasty (TKA) and to compare them with EP observed in total hip arthroplasty (THA), and primary knee and hip osteoarthritis (OA). DESIGN: Gene EP of TNF, IL-6, IL-8, CHIT1, BMP4, CCL3, CCL18, MMP9, RANKL, OPG, DC-STAMP and SOCS3 were assessed using quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) on tissues retrieved from patients with aseptically failed TKA (n = 21), THA (n = 41) and primary knee (n = 20) and hip (n = 17) OA. Immunohistochemistry was applied to localize the proteins. RESULTS: When compared to knee OA, the pseudosynovial tissue in TKA exhibit (1) elevation of alternative macrophage activation marker (CHIT1), chemokine (IL-8), and a proteolytic enzyme (MMP9); (2) downregulation of pro-inflammatory cytokine (TNF), osteoclastic regulator (OPG) and a stimulator of bone formation (BMP4); (3) no difference in IL-6, CCL3, CCL18, RANKL, DC-STAMP and SOCS3. The EP in TKA differed from EP in aseptically failed THA by lower CCL3 and DC-STAMP mRNA and protein expression. EP of all studied inflammatory and osteoclastogenic molecules were similar in knee and hip OA. CONCLUSIONS: Comparing to OA, aseptic loosening of TKA is associated with upregulated expression of CHIT1, IL-8 and MMP9, dysregulated RANKL:OPG ratio and low levels of inflammatory cytokines. Similar cytokine profiles were associated with primary knee and hip OA. Further research is required to explain the differences in CCL3 and DC-STAMP expression between failed TKA and THA.

[Patient with atypical neurocytoma -  case report]. / Pacientka s atypickým neurocytomem -  kazuistika.

BACKGROUND: Neurocytoma represents a rare tumor of the central nervous system usually slowly growing and generally with good prognosis after surgical resection with or without adjuvant radiotherapy. CASE: A 25-year- old woman presented with sudden fainting. During the initial workup, brain CT was completed with finding of tumor inside the third ventricle spreading into both lateral ventricles. The patient underwent partial surgical resection followed by radical gross resection, no adjuvant radiotherapy was indicated during the initial treatment and the patient was followed up with regular brain MRIs and clinical examinations. Thirty six months after the initial resection, there was progression on MRI and radiotherapy was recommended. At this moment, patient is 12 months after radiotherapy with stable disease on MRI and with good stable performance status. CONCLUSION: One of the greatest problems in the management of neurocytoma is the timing of adjuvant radiotherapy. From published data, it is clear that adjuvant radiotherapy increases local control; however, this has to be considered carefully against the possible risks from late side effects of radiotherapy considering long-time survival of the patients.

[The benefits of centralization of care for patients with acute upper gastrointestinal bleeding]. / Prínos centralizace péce o pacienty s akutním krvácením do horního trávicího traktu.

INTRODUCTION: Acute upper gastrointestinal bleeding still remains one of the serious conditions that require a rapid diagnostic and therapeutic intervention. Such procedure is highly dependent on good interdisciplinary cooperation, which, when centralized, may positively influence mortality and economic costs. OBJECTIVE: To determine the benefits of centralization of care provided to patients with acute bleeding in the upper gastrointestinal tract. PATIENTS AND METHODS: A total of 632 patients with acute bleeding were enrolled in the study at two different health-care establishments of the same type, however with a different organisation of care. We have evaluated the influence of the organisation of care on the length of hospitalization stay, mortality and economic costs. RESULTS: Centralized treatment significantly shortens the interval from hospital admission to endoscopic examination and leads to a reduction in mortality, although not statistically significant. On the other hand, it does not affect the length of hospitalization, the distribution of patients between internal and surgical departments, and provides no guarantee of lower economic costs. Many other factors play an important role in that respect. It seems, that the importance of centralization plays the main role especially in the pre-hospital and early hospital period, when it accelerates and simplifies the diagnostic and therapeutic procedure.

Differential expression of ABC transporters (MDR1, MRP1, BCRP) in developing human embryos.

Three ABC transporters (MDR1, MRP1, BCRP), belonging to the family of multidrug resistance (MDR) proteins, play a crucial role in the protection mechanisms during embryogenesis and mediate drug resistance in cancer cells. The distribution of these transporters in the series of human embryonal/fetal intestine, liver and kidneys of various stages of intrauterine development (IUD) by indirect two-step immunohistochemical method was investigated. The organ- and age-specific expression patterns of these transporters were depicted and compared with the expression in adult organs. The evaluation of intestine and liver samples demonstrate differences in expression pattern of ABC transporters during IUD. On the contrary, in kidneys the age-specific localization was not observed. However, the increasing positivity from the kidney surface towards deeper, more differentiated parts was found. Hopefully, our study may contribute to elucidation of the role of multidrug resistance (MDR) pathways during IUD in man.

[Prague Hepatology Meeting 2010--hepatopathology news]. / Prague Hepatology Meeting 2010--novinky z hepatopatologie.

This review summarizes the highlights in hepatopathology presented during the Prague Hepatology Meeting 2010. Gut flora (microbiome) and intestinal permeability seems to play an important role during progression into liver cirrhosis in patients with non-alcoholic steatohepatitis. Molecular diagnosis allows the distinction of several types of hepatocellular adenoma with different risks of malignant progression. Experimental hepatology is currently focused on the role of epigenetics during pathogenesis of alcoholic liver disease. The most studied topic in viral hepatitis C is the mechanisms of resistance to antiviral treatment. Pathogenesis of liver fibrosis and the role of keratins in the pathology of liver disease are areas of scientific interest as well.

Analysis of CD40 expression in breast cancer and its relation to clinicopathological characteristics.

Breast cancer is currently the most common cancer in women worldwide. For this reason, new biomarkers for better predicting response to treatment are needed. CD40, described as expressed in haematological and epithelial tumors, is linked to apoptosis and offers promise as a new predictive/ prognostic marker. We evaluated CD40 expression in formalin-fixed, paraffin-embedded samples from 181 breast carcinomas using immunohistochemical staining with CD40 antibody. Samples were divided according to hormone (oestrogen receptor /ER/, progesterone receptor /PR/) and her-2/neu status into groups: 1.Luminal A (ER+PR+her-2/neu-), 2. Luminal B (ER+PR+her-2/neu+), 3.Triple-negative (ER-PR-her-2/neu-) and 4. Her-2/neu (ER-PR-her-2/neu+). The results of CD40 staining were correlated with clinicopathological data. CD40 was found to be expressed in membrane, cytoplasm and nucleus. Normal ducts expressed cytoplasmic CD40 in 30% of cases, in breast tumor ducts in 53% of cases. CD40 was evaluated as an independent marker and significant positive correlation was found with Bcl-2 (p =0.002), early stage (p =0.016) and preoperative chemotherapy (p =0.043). There was higher overall survival for patients with cytoplasmic CD40 expression (0.05). Differences in expression of cytoplasmic CD40 between groups with different hormonal and her-2/neu status were statistically highly significant (p=0.00003). In groups with different hormonal status, a positive statistical correlation was found for the luminal A group with relapse (p=0.024) and stage (p=0.006). No correlation was found with age, disease onset, family history of cancer/ breast cancer, patient history, hormonal replacement therapy, menopausal status at onset of disease, adjuvant chemotherapeutic treatment or disease free survival. Nuclear expression of CD40 was found to be unrelated to any clinicopathological data. However, there was higher ratio of positive cases in cancer cases (83%) than in normal tissue (30%). In conclusion, cytoplasmic expression of CD40 is related to factors connected to better prognosis and suggest that CD40 may have potential as a new prognostic factor in breast cancer.

Replication stress and oxidative damage contribute to aberrant constitutive activation of DNA damage signalling in human gliomas.

Malignant gliomas, the deadliest of brain neoplasms, show rampant genetic instability and resistance to genotoxic therapies, implicating potentially aberrant DNA damage response (DDR) in glioma pathogenesis and treatment failure. Here, we report on gross, aberrant constitutive activation of DNA damage signalling in low- and high-grade human gliomas, and analyze the sources of such endogenous genotoxic stress. Based on analyses of human glioblastoma multiforme (GBM) cell lines, normal astrocytes and clinical specimens from grade II astrocytomas (n=41) and grade IV GBM (n=60), we conclude that the DDR machinery is constitutively activated in gliomas, as documented by phosphorylated histone H2AX (gammaH2AX), activation of the ATM-Chk2-p53 pathway, 53BP1 foci and other markers. Oxidative DNA damage (8-oxoguanine) was high in some GBM cell lines and many GBM tumors, while it was low in normal brain and grade II astrocytomas, despite the degree of DDR activation was higher in grade II tumors. Markers indicative of ongoing DNA replication stress (Chk1 activation, Rad17 phosphorylation, replication protein A foci and single-stranded DNA) were present in GBM cells under high- or low-oxygen culture conditions and in clinical specimens of both low- and high-grade tumors. The observed global checkpoint signaling, in contrast to only focal areas of overabundant p53 (indicative of p53 mutation) in grade II astrocytomas, are consistent with DDR activation being an early event in gliomagenesis, initially limiting cell proliferation (low Ki-67 index) and selecting for mutations of p53 and likely other genes that allow escape (higher Ki-67 index) from the checkpoint and facilitate tumor progression. Overall, these results support the potential role of the DDR machinery as a barrier to gliomagenesis and indicate that replication stress, rather than oxidative stress, fuels the DNA damage signalling in early stages of astrocytoma development.

[A different view of acute upper gastrointestinal bleeding in liver cirrhosis patients]. / Jiný pohled na akutní krvácení do horního trávicího traktu u pacientu s jaterní cirhózou.

OBJECTIVES: The study focuses predominantly on non-varicose sources of acute upper gastrointestinal bleeding in liver cirrhosis patients and aims to determine its mortality. METHODS AND SUBJECTS: The prospective examination included 137 liver cirrhosis patients with acute upper GIT bleeding. All the patients underwent an endoscopic examination. In case of multiple findings, defining the bleeding source was based on the specialist's attitude presented as the conclusion of the endoscopic examination. RESULTS: The most frequent causes of acute bleeding included oesophagus varices (57.7%), followed by peptic gastric and duodenal ulcers (18.2%), then portal hypertension gastropathy (9.5%), gastric varices (5.1%), reflux oesophagitis (2.9%), Mallory-Weiss syndrome (2.9%) and erosive gastropathy (1.5%). The endoscopy of the upper digestive tract resulted in a negative diagnosis in not more than 2.2% of patients. The majority of examinations showed multiple findings in the upper digestive tract, each of which could have been a potential cause of bleeding. Mortality in all bleeding cirrhotic patients reached 14.6%, 18.6% of which were related to the varicose type of bleeding and 7.8% to the non-varicose type. CONCLUSION: Portal hypertension led to bleeding (caused by varices and portal hypertension gastropathy) in 72.3% of patients, 62.8% patients suffered from purely varicose bleeding, 37.2% patients from non-varicose bleeding. Mortality was statistically significantly higher on 0.1 level of significance in cases of varicose bleeding in comparison with non-varicose bleeding. An emphasis should be laid on an early and detailed endoscopic examination leading to an appropriate diagnosis and therapy.

Analysis of VEGF, Flt-1, Flk-1, nestin and MMP-9 in relation to astrocytoma pathogenesis and progression.

Astrocytomas, particularly high grade astrocytoma, are brain tumors with potent angiogenic activity. Our immnunohistochemical study assessed vascular endothelial growth factor (VEGF), VEGF receptors (Flk-1, and Flt-1), the intermediate filamental protein nestin which plays a role in central nervous system development, and MMP-9, which belongs the family of matrix metalloproteinases implicated in tumor invasion and angiogenesis regulation. We investigated the expression of VEGF, its receptors, nestin and MMP-9 in astrocytomas and their correlation with tumor grade. We used paraffin-embedded samples from 66 patients, 29 with low grade (WHO-grade II) and 37 with high grade (WHO-grade III and IV) astrocytomas. Antibodies against VEGF, Flk-1, Flt1, nestin, CD34 and MMP-9 were used, followed by standard indirect immunohistochemical methods. Expression of Flt-1 and Flk-1 showed no significant differences between low and high grade tumor groups. Expression of VEGF and MMP-9 was increased in the high grade group (p equal to or less than 0.026 and 0.024). Nestin expression in tumor astrocytes and endothelial cells increased in high grade group (p same 0.007 and 0.003). Higher expression of VEGF in high grade astrocytomas may subsequently lead to activation of survival, angiogenesis and migration. Expression of nestin and MMP-9 also suggest their likely role in astrocytoma vascular development and proliferation.

The Role of PPARs in MDR - a lesson from embryonic development.

One of the most important features of embryonic cells is their resistence to xenobiotics, which provides a natural protection for embryos against these potentially harmful molecules. In this way, embryo cells resemble cancer cells and thus understanding the basis of this phenomenon may contribute to overcoming the multi-drug-resistance (MDR) of some tumours. Peroxisome proliferator-activated receptors (PPARs) are steroid nuclear receptors that regulate diverse biological processes such as lipid and carbohydrate metabolism, development, differentiation, apoptosis, neoplastic transformation, inflammation and regeneration of tissues. Recently it has been found that they may also regulate the expression of some MDR proteins. In this article we summarise the main known relationships between some MDR pumps and three isoforms of PPAR receptors (PPAR-alpha, PPAR-beta/delta, PPAR-gamma). We hypothesize that regulation of MDR proteins by PPAR ligands in embryos could lead to the improvement of cancer treatment.

Expression of STATs and their inhibitors SOCS and PIAS in brain tumors. In vitro and in vivo study.

Proteins of STAT family belongs to the transcription factors. Through their binding to the DNA specific sites and consequent regulation of transcription of various genes, these signaling proteins play an important role in many cell functions. Recent studies demonstrated persistent activation of STATs and loss of their natural inhibitors SOCS and PIAS in various human cancers. There is also evidence that experimental pharmacologic or genetic modulation of their function mignt by a new approach in anticancer treatment. The aim of this study was in vitro assesment and analysis of expression of STATs, SOCS and PIAS in glioblastoma cell lines undergoing treatment by PPARgamma agonists/antagonists because PPARgamma and STATs are tightly regulated by an overlapping set of nuclear regulatory proteins. We further analysed immunohistochemical expression of these proteins in vivo, with its correlation to grading in various brain tumors. The results of in vitro study showed decreased expression of phosphorylated form of STAT3 and increase of its inhibitors SOCS3 and PIAS3 in glioblastoma cell lines after treatment with IC50 of PPARgamma agonist ciglitazone. In vivo study failed to reveal changes in STAT3 and SOCS3 expression in either low and high grade astrocytomas, however we detect lower expression of STAT2 in low grade astrocytomas when comparing with high grade astrocytomas and lower expression of STAT3 in ependymomas when comparing with anaplastic ones. The results showed existing relationship between STAT and PPARgamma signaling in glial tumors and further suppport expected important role of STATs in regulation of growth and differentiation in these tumors.

[Reminiscences on the occasion of the Whipple's disease centenary: a summary paper]. / Reminiscence k stoletému výrocí objevu Whippleovy nemoci: souborný referát.

The disease referred to eponymically as Whipple's disease (WD) in medical literature was thoroughly described by the American physician and pathologist George Hoyot Whipple (1878-1976) in 1907 and given a temporary denomination of "intestinal lipodystrophy". According to literature, WD is rare, but its precise incidence has not yet been established. Familial incidence of the disease is acknowledged, and its immunogenetic pathogenesis is assumed. The incidence ofWD is prevailingly observed in middle-aged men (mean age 55), exceptionally at child age - the ratio being 3 to 6 for men and women, respectively. 1. Clinical diagnosis is based on symptoms in the GIT region and, in rare cases, on extraintestinal symptoms. Clinical symptomatology includes: abdominal pain with persistent diarrhoea (steatorrhoea), symptoms typical of malabsorption connected with weight loss, fevers, polyarthritic symptoms, swollen lymph nodes and, in part of patients, skin hyperpigmentation. Anaemia and hypoalbuminaemia (reduced IgA) are typically detected in laboratory tests. Rarer extraintestinal symptoms of the disease are of a diverse nature: cardiac lesions, cerebral lesions, ocular symptoms, conspicuous or even tumour-like enlargement of lymph nodes, lesions of the hemopoietic system. The clinical course ofWD is of progressive or remittent nature and the disease is fatal without treatment. Long-term therapy with antibiotics, especially a combination oftetracyclines (doxycyclin) and corticoids (dexametazone), or chloramphenicol in case of cerebral lesion, have a significantly positive effect on the course and prognosis of WD. From the point of view of pathology, WD is a multisystem infectious disease (Tropheryma whipplei) primarily affecting the GIT (39, 47, 52, 103) or different extraintestinal locations. Due to the known diversity of clinical symptoms, no clinical-diagnostic standard has been established for WD. Differential diagnosis includes different multisystem diseases, primarily malignant lymphomas (especially Hodgkin's disease). From the pathogenetic point of view, we can either assume the effect of a particular cytokine (or TNFalpha) activating macrophage phagocytosis or, if its production is normal, a disorder or defect of the respective receptor in the macrophage cellular membrane. The identification of "Whipple's bacteria" - Tropheryma whipplei - gen. nov. et sp. nov. was made possible by modern molecular biology research methodologies. Its cultivation allows both for the acquisition of the specific antibody and of detailed knowledge of its genoma (PCR, 16S rRNA sequencing).

Re-expression of nestin in the myocardium of postinfarcted patients.

Intact cardiac muscle cells in the adult heart do not express intermediate filament nestin. In this study, we report on widespread expression of intermediate filament nestin in human myocardium of patients who died from the myocardial infarction. Nestin was detected in cardiomyocytes, endothelial cells, and few interstitial cells. Elevated levels of nestin were observed in cardiac muscle cells in all specimens, although the intensity of immunoreactivity and distribution of the signal differed. The strongest immunoreactivity was observed from 4 days after myocardial infarction in the infarction border zone where nestin was distributed homogeneously in the entire sarcoplasm of cardiac muscle cells. Within the following week, nestin in immunoreactive cardiomyocytes was redistributed and restricted to small subsarcolemmal foci and to intercalated discs. Angiogenic capillaries that grew between vital nestin-positive cardiomyocytes and entered the necrotic area expressed also high levels of nestin. Nestin-positive endothelial cells were often observed in mutual interactions with nestin-positive cardiac muscle cells. These findings document a crucial role of nestin in remodeling cytoskeleton of cells in the human postinfarcted myocardium.

[Current concepts and morphological aspects of drug induced liver injury]. / Lékové postizení jater--soucasný stav problematiky se zamerením na morfologický obraz.

Currently, the following areas are in the focus of interest in hepatology: study of hepatic stem cells, mechanisms of fibrogenesis and cirrhosis development; mechanisms of the development of steatosis, steatohepatitis and NASH/NAFLD; rejection of liver transplants; diagnosis of autoimmune hepatitis, primary sclerosing cholangitis and primary biliary cirrhosis; diagnosis of hemochromatosis; the importance and relevance of grading and staging of chronic hepatitis. DILI (Drug Induced Liver Injury) is also in the focus of interest. DILI is important because the histology of DILI overlaps almost all areas of liver pathology unrelated to drug injury. Moreover, there is not unified approach to DILI diagnostic and references are almost solely case reports. The diagnosis of DILI is also complicated by lack of information on pharmacological anamnesis of patients, frequent combination of drugs with natural compounds and influence of environmental factors. DILI is a cause of 10% of acute hepatitis and 15 % of liver failures with subsequent transplantation. Hepatotoxicity is also one of the most important reasons for drug withdrawal. This review covers basic informations on classification of hepatotoxic drugs, on the system of liver biopsy assessment when DILI is suspected, and it is an overview of the main histologic pictures of liver biopsy with examples of the most important drugs and differential diagnosis.