[Adverse effects of modern treatment of malignant melanoma and their treatment/ management]. / Nezádoucí úcinky novodobé lécby maligního melanomu a jejich lécba/ management.

BACKGROUND: Malignant melanoma belongs to the most deadly human tumours and despite all preventive programs its incidence continues to rise. Until 2011, chemotherapy was the only therapeutic option for inoperable or metastatic disease in the Czech Republic. However, new treatment modalities (e. g. targeted therapy) have been introduced recently. AIM: Since most of the modern drugs are still available only in clinical trials, the aim of this article is to provide a brief and comprehensive review of current treatment options for metastatic disease. The attention is focused on their potential side effects, so that doctors who do not usually deal with these drugs would get acquainted with them. This could contribute to a prompt management of associated symptoms or an early referral of the patient to an appropriate clinical centre without undue delay.

[Vascular endothelial growth factor in ovarian cancer patients]. / Cévní endotelový rustový faktor u pacientek s karcinomem ovaria.

OBJECTIVE: To compare plasma VEGF (vascular endothelial growth factor) levels in ovarian cancer patients, in patients with benign ovarian tumors and healthy women. DESIGN: Prospective study. SETTING: Department of Gynecology and Obstetrics, Medical Faculty Charles University, Prague and University Hospital, Hradec Králové. Department of Immunology and Alergology, Medical Faculty Charles University, Prague and University Hospital, Hradec Králové. METHODS: VEGF was estimated by ELISA (R&D Systems). RESULTS: We found that plasma VEGF levels were associated with the International Federation of Gynecology and Obstetrics (FIGO) stage (FIGO I+II, n=8) Med = 425,53 pg/ml (range 142,30-982,40 pg/ml), (FIGO III+IV, n=29) Med = 941,48 pg/ml (range202,10-2857,80 pg/ml) (p=0,03). Patients with primary ovarian cancer (n=37) had a significantly higher plasma VEGF level Med = 829,93 pg/ml (range142,30-2857,80 pg/ml), compared with patients with benign ovarian tumors (n=15) Med = 426,28 pg/ml (range 32,00-922,20 pg/ml) and healthy women (n=21) Med = 283,13 pg/ml (range 80,50-735,20 pg/ml) (p=0,0003). VEGF levels were lower in plasma (n=79) Med = 575,49 pg/ml (range 55,80-2185,00 pg/ml) compared with VEGF levels in ascitic fluid (n=37) Med = 745,74 pg/ml (range 142,30-2185,00 pg/ml) (p=0,04) in ovarian cancer patients. CONCLUSION: Plasma VEGF assay before primary treatment and the changes during the other treatment should contribute to better understanding of angiogenesis in ovarian cancer patients. Plasma VEGF correlates with the stage of primary ovarian cancer.

[Changes in immune reactivity in cancer patients]. / Zmeny imunologické reaktivity u onkologických pacientu

It appears that the long-accepted paradigm that cytostatic and radiation therapy cause only immunosuppression, is not so clearly true. With regard to new knowledge in cancer immunology field, it seems that not only cytostatic and radiation therapy plays an important role in the alteration of immune system. There are many other factors influencing immunity like tumour environment itself, the use of immunomodulatory drugs or even the mental condition of cancer patients. The aim of review is to familiarize physicians with possible alterations of the immune system in cancer patients.

Black mamba dendroaspis polylepis bite: a case report.

Although very rare in a European context, a bite from the black mamba Dendroaspis polylepis is an event that poses an immediate threat to life. Given the content of neurotoxins in the snake's venom, the mortality of envenomation reaches 100% in almost every case if ventilation is not provided in a timely manner and adequate therapy initiated. The report describes a case of a snake breeder being envenomed. This 31-year-old man was bitten by a black mamba on his finger, and who subsequently developed clinical symptoms of envenoming typical for the species. Thanks to mechanical ventilation being employed promptly, with myorelaxation during generalized muscle fasciculations, and particularly owing to the eventual antivenin therapy, the patient's condition settled without consequences. In addition to describing the given case in detail, the paper discusses the composition and mechanisms of action of black mamba venom, while providing guidelines for adequate therapy.

No evidence for linkage between the hereditary angiooedema clinical phenotype and the BDKR1, BDKR2, ACE or MBL2 gene.

Hereditary angiooedema (HAE) is a life-threatening disease with poor clinical phenotype correlation with its causal mutation in the C1 inhibitor (SERPING1) gene. It is characterized by substantial symptom variability even in affected members of the same family. Therefore, it is likely that genetic factors outside the SERPING1 gene have an influence on disease manifestation. In this study, functional polymorphisms in genes with a possible disease-modifying effect, B1 and B2 bradykinin receptors (BDKR1, BDKR2), angiotensin-converting enzyme (ACE) and mannose-binding lectin (MBL2), were analysed in 36 unrelated HAE patients. The same analysis was carried out in 69 HAE patients regardless of their familial relationship. No significant influence of the studied polymorphisms in the BDKR1, BDKR2, ACE and MBL2 genes on overall disease severity, localization and severity of particular attacks, frequency of oedema episodes or age of disease onset was detected in either group of patients. Other genetic and/or environmental factors should be considered to be responsible for HAE clinical variability in Caucasians.

[NK cells, chemokines and chemokine receptors]. / NK bunky, chemokiny a chemokinové receptory.

NK cells are an important component of natural immunity, which provides a defence response against viruses, bacterial and parasitic intracellular pathogens and tumour cells. NK cells are capable of rapid responses without prior sensitization and cytotoxic response is independent of the presence of the antigens of the major histocompatibility system. NK cells produce a number of cytokines (e.g. INF-gamma, GM-CSF and TNF-beta) and chemokines and in this way they regulate both the natural and acquired immune response. By contrast, NK cells are regulated both positively and negatively by cytokines and chemokines produced by other immune cells. Attention is focused on the possibility of influencing the tumour process by using cytokine- and chemokine-activated NK cells. In studies in mice models as well as in several clinical trials, it has been shown that the presence of cytotoxic cells in tumour stroma is associated with a more favourable prognosis of cancer.There is also plenty of evidence that in tumour stroma a number of cytokines and chemokines are produced which may have ambivalent effects.

[Inflammatory reaction and its importance in the course of tumour process]. / Zánetlivá reakce a její význam v prubehu nádorového procesu.

This article gives basic information about the formation and the course of the inflammatory reaction, which is the important adaptation mechanism of the organism on the various external and internal injurants. The main attention is fixated on the role of the immune system in the course of the inflammatory reaction. It deals with the presence and the sense of the inflammatory reaction in the tumour environment in more detail. It gives the short information about the mechanisms of the antitumour defence. The influence ofthe inflammatory reaction on the genesis, the course and the progression of the tumour disease is discussed in the third chapter.

Determination of angiogenic factors in serum by protein array in patients with renal cell carcinoma.

Using the protein array method we determined the serum levels of a number of angiogenic factors. We identified serum levels of angiogenin, PDGF and MCP-1 (CCL2 chemokine) in serum of 32 patients with RCC, and 14 healthy volunteers by means of antibody array analysis. The patients were divided into three groups according to their disease stages (I+II, III, and IV). We found significant differences between the controls and patients with RCC both pre-operatively and post-operatively in angiogenin, PDGF and MCP-1 serum levels. The increase in angiogenin, PDGF and MCP-1 lasted in patients with RCC stages I-III even without metastases eight weeks post-operatively. The patients with stage IV RCC showed disturbed production of PDGF and MCP-1. Protein array analysis is a powerful tool for the identification of large numbers of trace proteins. Multiplex antibody array is able to provide data more precisely reflecting the nature of pathological processes.

[Monitoring of anti-tumour cell-mediated response in patients with renal cell carcinoma, disturbance of T cell proliferation]. / Sledování protinádorové bunecné imunitní odezvy u nemocných s renálním karcinomem, porucha proliferace T-lymfocytu.

INTRODUCTION: When checking tumour growth, a number of observations indicate that the immune system plays a significant role in patients with renal cell carcinoma (,,RCC"). Infiltration by lymphocytes (tumour infiltrating lymphocytes, "TILs") is more prevalent in RCC than any other tumours. T lymphocytes are the dominant population of TIL cells. Views concerning the role ofT lymphocytic subpopulations, B lymphocytes and NK cells in an anti-tumour response are not established. AIM: The aim is to determine the phenotype and activation of lymphocytic cells and to compare their representation in tumour stroma (TIL), peripheral blood (PBL) and renal vein blood in patients with RCC. PATIENTS AND METHODS: The samples of peripheral blood taken from the cubital and renal veins and tumour stroma cells were obtained from 60 patients in the course of their surgeries carried out due to primary RCC. TILs were isolated from mechanically disintegrated tumour tissue. Immunophenotype multiparametric analysis of PBL and TILs was carried out. Their surface and activation characteristics were determined by means of flow cytometer. RESULTS: CD3+ T lymphocytes (70.4%) were the main population of TILs. The number of CD3+/CD8+ T lymphocytes was significantly higher in TILs, 39.7% (p < 0.01), while CD4+ T lymphocytes were the majority population in peripheral blood, 41.35% (p < 0.001). The representation of CD3+/69+ T lymphocytes was significantly higher in TILs, 32.05%, compared to PBL (p < 0.001). On the contrary, the numbers of CD3+/CD25+, CD8+/57+ and CD4+/RA+ (naive CD4+ T lymphocytes) were higher in PBL (p < 0.001). The differences in representation of (CD3+/16+ 56+) NK cells and CD3+/DR+ T cells in TILs and PBL were not significant. CONCLUSION: The above-mentioned results prove that the characteristics and intensity of anti-tumour responses are different in compared compartments (tumour/PBL). CD3+/CD8+ T lymphocytes are the dominant lymphocytic population of TILs. The knowledge of phenotype and functions ofeffector cells, which are responsible for anti-tumour response, are the basic precondition for understanding the anti-tumour immune response and the cause of its failure.

Genetic defects in common variable immunodeficiency.

Common variable immunodeficiency (CVID) is the most frequent clinically manifested primary immunodeficiency. According to clinical and laboratory findings, CVID is a heterogeneous group of diseases. Recently, the defects of molecules regulating activation and terminal differentiation of B lymphocytes have been described in some patients with CVID. In this study, we show the overview of deficiencies of inducible costimulator, transmembrane activator and calcium-modulator and cytophilin ligand interactor, CD19 molecules, their genetic basis, pathogenesis and clinical manifestations.

[Late complications of chronic respiratory infections in patients with common variable immunodeficiency]. / Pozdní komplikace chronických zánetu respiracního traktu u nemocných s beznou variabilní imunodeficiencí.

Common variable immunodeficiency (CVID) is the most frequent serious humoral deficiency manifested in adulthood in the form of acute and chronic respiratory infections which in most patients lead to respiratory failure. Retrospective analysis of 28 CVID patients was made. Mean age at time of diagnosis was 38.6 +/- 18.6 years. The time which elapsed from the first symptoms to determining the diagnosis was three times as long as that indicated in other studies, i.e., 14.1 +/- 10.2 years. Twenty-three patients (82.1%) had respiratory complications. Chronic obstructive pulmonary diseases, n=16 (57.1%), and bronchiectasis, n=10 (35.7%), were the most frequent types of lung damage. In addition, two patients (7.1%) displayed evidence of interstitial lung process. Morbidity associated with CVID can be reduced by early diagnosis and adequate dosage of immunoglobulins to minimise the occurrence and progression of lung damage.

[Tumor angiogenesis]. / Nádorová angiogeneze.

Angiogenesis have shown a major role in tumor growth and metastasis formation. For tumor growth beyond the size 1-2 mm3, angiogenesis must be started to form vascular supply of tumor cells. Angiogenesis is a complex process, involving degradation of the basement membrane of preexisting vessel, proliferation of endothelial cells towards the angiogenetic stimulus, maturation of endothelial cells with formation of luminized capillary, and finally formation of a functional vessel, surrounded by basement membrane and pericytes. Angiogenesis is regulated by numerous angiogenic and anti-angiogenic factors. Hypoxia is a significant stimulus for angiogenesis. For many cancers the extent of vascularisation is a negative prognostic indicator signifying aggressive disease and increased potential for metastasis.

Campylobacter sepsis with multiple organ failure in IgG subclass deficiency.

Some patients with immunodeficiency develop clinical features of autoimmune disorders. A previously asymptomatic antibody deficiency can underlie the development of autoimmune diseases and a severe course of infection, with a risk of sepsis; such cases are known in selective IgA deficiency. On the other hand, little information is available on selective IgG subclass deficiencies. An unexpectedly severe course of Campylobacter infection in a 19-year-old woman with a previously undiagnosed complex immune disorder, including selective IgG1 immunodeficiency, Hashimoto's autoimmune thyroiditis with hypothyroidism combined with Addison's disease presumably due to autoimmune adrenalitis, autoimmunity and allergy is described. The pathophysiological mechanisms of autoimmunity in latent humoral defects are discussed.

[Latex allergy--report on two cases]. / Alergie na latex--dva kazuistické prípady.

In the last few years latex allergy has been increasingly recognised as a potential medical problem because of the increase in frequency and potential severity of latex-mediated reactions. Latex allergy is an IgE-mediated hypersensitivity response to natural rubber latex protein with a variety of clinical signs ranging from contact urticaria, angioedema, asthma, and anaphylaxis. Also IV. type of immunology response can participate in it. In highest risk for latex allergy are patients with spina bifida, but health care workers, latex industry workers, patients with multiple surgical procedures and others who wear gloves are also at risk. Patients with history of atopy belong to the high risk group. Diagnosis is done by positive in vitro tests (EAST, CAP-FEIA, immunoblott etc.) and skin prick test. Allergen avoidance and substitution and the use of latex-safe devices including synthetic gloves are essential for the affected patient.

[Multidrug resistance markers monitoring in acute myeloid leukemia cells]. / Sledování vybraných znaku mnohocetné rezistence k cytostatikum u leukemických bunek pacientu s diagnózou akutní myeloidní leukémie.

BACKGROUND: P-gp, MRP, LRP, Bcl-2, and Bax proteins play an important role in the multidrug resistance of leukemic cells. P-gp, MRP, and LRP proteins decrease the intracellular drug concentration, Bcl-2 and Bax proteins influence the apoptosis process. The overexpression of some of these proteins is associated with poor prognosis of leukemia. The aim of this study was to find the relationship between some multidrug resistance markers and some clinical and laboratory parameters. We compare also P-gp expression between acute myeloid leukemia (AML) FAB subgroups M0-M6. METHODS AND RESULTS: With use of flow cytometry we measured the expression of P-gp, MRP, LRP, Bcl-2, and Bax proteins in acute myeloid leukemia patients cells. The study proved the association between blast percentage and P-gp protein expression (p=0.015) and the association between blast percentage and Bcl-2 protein expression (p=0.041). It was also shown the tendency of the LRP protein expression to associate with higher age of patients (p=0.062). Another correlation between MRP and Bax expression (p=0.006), as well as between LRP and Bax expression was found (p=0.034). In AML M0 FAB subgroup of patients the trend to higher P-gp protein expression was observed. CONCLUSIONS: The laboratory methodology to multidrug resistance markers detection was introduced. Some relations between multidrug resistance markers playing role in acute myeloid leukemia patients prognosis were suggested.

[Immunopathogenesis of diabetes mellitus I]. / Imunopatogeneze diabetes mellitus I. typu.

The pathogenesis of diabetes mellitus I is multifactorial. Both genetic predisposition and environmental factors are involved in the deregulation of immune system with subsequent breakage of self-tolerance. Autoantibodies against numerous autoantigens are raised during this long-term process but the principal pathogenic mechanism is cytotoxic killing of beta cells of pancreas mediated predominantly by T cells. There could be the chance to interrupt this pathogenetic process by manipulation with immune system to achieve induction of tolerance. Such clinical trials are now under progress.

[Importance of selected laboratory indicators in the differential diagnosis and monitoring of multiple myeloma]. / Význam vybraných laboratorních ukazatelu pro diferenciální diagnostiku a sledování aktivity mnohocetného myelomu.

Multiple myeloma is one of the most common haematologic malignancies. Currently there are numerous studies looking for new prognostic markers in multiple myeloma. The most important of them are the markers related to proliferative activity of neoplastic cells or to size of tumor mass. The subject of this paper are the results obtained from investigation of some such laboratory markers in a group of patients with monoclonal gammopathies diagnosed at our department in the last 3 years. We analyzed blood and bone marrow samples from 51 patients with new diagnosed monoclonal gammopathies, 14 of them were patients with monoclonal gammopathy of undetermined significance and 37 patients had multiple myeloma. 17 patients with multiple myeloma were treated by high-dose chemotherapy regimen. We assessed significance of selected laboratory markers for differential diagnosis of monoclonal gammopathies and for monitoring of activity of multiple myeloma. Among the investigated parameters, we verified the significance of cell cycle analysis of bone marrow plasmatic population and of the determination of the number of circulating myeloma cells in differential diagnosis of monoclonal gammopathies. In our opinion, the determination of soluble CD138, beta 2-microglobulin and neopterin serum levels can be also recommended as helpful markers for a solution of this problem. Except of beta 2-microglobulin serum level we did not find statistically significant correlation with activity of multiple myeloma in any of the investigated parameters.

The phenotype of ascitic fluid lymphocytes in patients with ovarian carcinoma and other primaries.

BACKGROUND: Ascitic tumor-infiltrating lymphocytes (TIL) are a potential source of effectors for adoptive immunotherapy. PATIENTS AND METHODS: The TIL phenotype was examined by two-color flow cytometry in malignancy-related ascites of 49 patients with different primaries. Interleukin-10 (IL-10) and neopterin were determined in ascitic fluid by enzyme-linked immunoassay. RESULTS: Malignant melanoma patients had significantly higher CD3(+), CD3(+)CD8(+) and CD3(+)CD95(+), and lower CD3(+)CD4(+) lymphocyte numbers than patients with other primaries. Ovarian cancer patients had higher CD3(+)CD45RO(+), CD8(+)CD28(+), CD19(+)CD86(+), CD19(+) and CD19(+)CD86(+) lymphocyte numbers, and lower NK cell numbers than patients with gastrointestinal and pancreatic primaries. Pretreated patients had significantly lower concentrations of IL-10, lower CD8(+)CD28(+), CD3(+)CD45RA(+), and higher CD3(+)CD80(+) numbers than chemotherapy-naïve patients. Patients with hepatic metastases had lower CD3(+), CD3(+)CD4(+) and CD3(+)CD45RO(+), and higher CD3(+)CD25(+) and NK cell numbers than patients without liver metastases. A substantial number of cells exhibited dendritic cell phenotype. Significant correlations were observed between neopterin and IL-10 concentrations, and numbers of CD8(+)CD28(+) and CD3(+)CD80(+) lymphocytes. CONCLUSIONS: Some parameters of TIL phenotype differ depending on primary, previous treatment, or the presence of liver metastases. A negative correlation was observed between IL-10 and neopterin, and an opposing effect of local concentrations of IL-10 and neopterin on the numbers of CD8(+)CD28(+) and CD3(+)CD80(+) was noted.

The peripheral blood leukocyte phenotype in patients with breast cancer: effect of doxorubicin/paclitaxel combination chemotherapy.

Presence of functional immune system is critical for any attempt aimed at improving survival of breast cancer patients by strategies based on immune system manipulation. We evaluated by flow cytometry the phenotype of peripheral blood leukocyte of 43 breast cancer patients. In 11 patients, the phenotype was evaluated before and during the chemotherapy by combination of doxorubicin and paclitaxel (AT). Compared with controls breast cancer patients had significantly higher relative and absolute numbers of CD3 HLA-DR+, CD3+CD69+ and CD14+CD16+, and significantly lower percentages of CD3 and CD8+CD28+ cells. After one cycle of AT, the absolute numbers of CD3 , CD3+CD4+, CD3+CD8+ and CD8+CD28+ cells increased significantly. Present data show a presence of T-cell activation in breast cancer patients. Administration of AT may lead to an increase in functional T-cells in peripheral blood, indicating a potential for combining chemotherapy with immunotherapy in the treatment of breast cancer patients.

[Multidrug resistance of leukemia cells to cytostatics--the functional fluorescent dye efflux assay evaluated by flow cytometry]. / Mnohocetná rezistence leukemických bunek k cytostatikum--funkcní testy akumulace fluorescencních barviv hodnocené prutokovou cytometrií.

The functional fluorescent dye efflux assays are used in the study of the multidrug resistance of the malignant cells to the cytotoxic drugs which is caused by the overexpression of P-glycoprotein (P-gp) or another membrane transport system--a multidrug resistance related protein (MRP). P-glycoprotein and a multidrug resistance related protein are involved in the efflux of cytotoxic drugs out of the cell and are responsible for the resistance. Fluorescent dye efflux mediated by these proteins could be evaluated by the flow cytometry. This test seems to be an optimal approach to study the multidrug resistance. With help of the specific inhibitors such as verapamil and cyclosporine A, the functional capacity of these proteins and the possibility to overcome the multiresistant phenotype can be revealed. The cell line K562 with transfected P-glycoprotein gene serves as a model system for the studying of the transport function with the use of the fluorescent substrates and P-glycoprotein inhibitors.