Perspective in predicting the effect of pleurodesis in the treatment of malignant pleural effusions.

BACKGROUND: Pleurodesis with talc has been used for many years. No objective criteria of inflammatory symptoms are known in order to use them to predict the effect of therapy and selection of patients. PURPOSE OF THE STUDY: To objectify the course of local inflammatory changes in the pleural cavity, quantify their dynamics in a clinically used procedure, and to determine specific predictors of inflammation as perspective markers of selection of patients suitable for talcage. MATERIAL AND METHOD: A total of 114 patients were retrospectively divided into Group A (n1 = 98) of patients without relapse and Group B (n2 = 16) of patients with relapse of exudate. The need for repeated thoracic punctures or drainages over the course of a 12-month monitoring period was the criterion of treatment failure. RESULTS: The groups were not different as for the baseline values of sTLR-2 (p0 = 0.638). Group A showed a marked growth during the monitoring period in 2 hours following talcage (p2= 0.002) and in 24 hours (p24 = 0.016). Group B showed higher sCD-163 levels (p0 < 0.001). The initial sTREM-1 values and those after two hours were similar in both groups (p0 = 0.146; p2 < 0.0641). In 24 hours, Group A had markedly higher values (p24 < 0.001). CONCLUSION: The sTLR-2, sCD-163 and sTREM-1 values can be prospectively determined. High sCD-163 values predict unsuitable selection of a candidate for talcage. The degree of inflammatory response can be evaluated using sTLR-2 or sTREM-1. Talcage using an inserted thoracic drain can be repeated at low levels. The cost of ELISA examination is approximately 18 euros (Tab. 1, Fig. 4, Ref. 20).

Impact of methylprednisolone in priming solution of cardiopulmonary bypass on anti-inflammatory CD163 receptor during cardiac surgery.

We evaluated the influence of methylprednisolone in cardiopulmonary bypass fluid on scavenger receptor for hemoglobin CD163 molecule expression on monocytes of patients who underwent elective coronary artery bypass grafting with cardiopulmonary bypass with either exposure to methylprednisolone present in the cardiopulmonary bypass fluid (20 patients), or without methylprednisolone in the cardiopulmonary bypass fluid (22 patients) and operated on without cardiopulmonary bypass (42 patients). The dynamics of CD163 expression was also followed in patients operated on without cardiopulmonary bypass. This study was a retrospective analysis of a comparison of two studies. The expression of CD163 was determined quantitatively by standardized flow cytometry technique. The similarities in the dynamics of CD163 monocyte expression, comparing the patients operated on with or without cardiopulmonary bypass, were found. Compared to the preoperative level, CD163 monocyte expression was significantly elevated on the 1(st) postoperative day. Monocyte CD163 expression on the 1(st) postoperative day was evidently similar in both groups of patients operated without cardiopulmonary bypass (median value of mean fluorescence intensity (MFI) 18,896; interquartile range from 27,538 to 57,711; median value of MFI 18,863; interquartile range from 16,514 to 26,559; n.s.), suggesting high reproducibility of our flow cytometric method; the monocyte CD163 expression was significantly higher (median value of MFI 37,902; interquartile range from 27,538 to 57,711) on the 1(st) postoperative day in patients exposed to methylprednisolone compared to patients without this exposure (median value of MFI 20,995; interquartile range from 16,321 to 29,623) (p<0.001). We concluded that the expression of hemoglobin scavenger receptor CD163 on monocytes of cardiac surgical patients is induced by methylprednisolone present in cardiopulmonary bypass fluid.

Interferon gamma receptor expression on granulocytes of cardiac surgical patients is modulated differently by the type of cardiopulmonary bypass used.

AIMS: To follow the IFNγ receptor expression on monocytes and granulocytes of cardiac surgical patients with respect to the type of cardiopulmonary bypass (CPB). METHODS: Expression of IFNγ receptor on monocytes and granulocytes of 26 cardiac surgical patients operated with the use of either "standard" or "miniaturised" CPB was determined by flow cytometry. RESULTS: The significant increase in IFNγ receptor expression on monocytes on the 1(st) and on the 3(rd) postoperative days was revealed in both groups of patients (p<0.001) irrespective of the type of CPB used, being non-significantly different between groups. In contrast, the expression of IFNγ on granulocytes displayed significant differences in terms of the CPB used. Whereas, in "standard" CPB patients, granulocyte INFγ receptor expression reached its maximum immediately after surgery (p<0.01), in "miniivasive" CPB patients, the peak in INFγ receptor expression was postponed to the 1(st) postoperative day (p<0.05). Statistically significantly higher IFNγ receptor expression on granulocytes was found in "standard" CPB patients (p<0.05). CONCLUSION: Compared to "miniaturised" CPB patients, the significantly higher IFNγ receptor expression on granulocytes was found in "standard" CPB patients (p<0.05) on the 1(st) postoperative day.

Quantification of ZAP-70 expression in chronic lymphocytic leukemia: T/B-cell ratio of mean fluorescence intensity provides stronger prognostic value than percentage of positive cells.

Expression of ZAP-70 measured by flow cytometry belongs to the most powerful prognostic parameters in chronic lymphocytic leukemia (CLL). However, many technical factors such as setting of the positivity threshold may significantly influence results.. Quantification using mean fluorescent intensity (MFI) may eliminate the subjective error which is inevitable in the isotype control method. The aim of the present project was therefore to assess the prognostic significance of ZAP-70 using three different methods. Between 2005 and 2010 we measured ZAP-70 expression in 157 patients with CLL (108 males, 49 females, median age 60 years [range, 31-82]; low/intermediate/high Rai risk in 41/48/11%). Expression of ZAP-70 was determined by flow cytometry using phycoerythrin (PE)-conjugated monoclonal antibody, clone 1E7.2. Evaluation was performed by 1) percentage of positive cells compared to isotype control (cut-off 20%), 2) MFI ratio of T-cells/CLL cells (cut-off 3.0); 3) MFI ratio of ZAP-70/isotype control on CLL cells (cut-off 2.5). MFI method with T-cells/CLL cells ratio was the best in the identification of patients with unfavourable outcome: ZAP-70 positive patients had significantly shorter time to treatment (TTT, median 24 vs. 55 months, p=0.0001) and overall survival (OS, median 97 vs 174 months, p=0.0074). The differences in TTT a OS were not significant with the use of isotype percentage and MFI isotype methods. Combined analysis of ZAP-70 with CD38 expression or IgVH mutation status lead to identification of a subgroup with the longest TTT and OS (ZAP-70 and CD38 negative, p<0.0001 and p=0.012; ZAP-70 negative and mutated IgVH genes, p<0.0001 and p=0.0019). In conclusion, our results suggest that measurement of ZAP-70 expression in CLL by MFI using T-cells/CLL cells ratio might be the optimal method for accurate prediction of clinical course. Combined analysis of ZAP-70 with CD38 or IgVH mutation status further refined individual patient´s prognosis.

[Monitoring the effectiveness of surgical treatment of malignant pleural effusions]. / Monitorace efektivity chirurgické lécby maligních pleurálních výpotku.

BACKGROUNDS: Recurrent pleural effusion occured in 10% of cancer patients. Repeated thoracocentesis or drainage may be complicated by pneumothorax, haemothorax or pleural cavity infection. Thoracoscopic talc poudrage is one of the most effective pleurodesis technique in patients with malignant pleural effusions. The effectiveness of such pleurodesis is reaching the 70 to 90%. This surgical approach also allows to take a biopsy for histological verification of the process. An effort to predict the success rate of chemical pleurodesis on the basis of the body's general inflammatory reaction rate, which is determined by the dynamics of values of humoral and cellular inflammatory parameters in both serum and in pleural effusion. PATIENTS: In the period 6/2008-12/2009 we applied biotalcum to 14 patients with malignant pleural effusions. The group of patients consisted of 10 male patients and 4 female patients of average age 71 years. Indication to include patients in the group was a second or further thoracic puncture, shortening of the interval between interventions, estimated time of survival > 3 months and the possibility of operation under selective pulmonary ventilation. METHOD: We performed the collection of pleural effusion and blood serum at 12-hour intervals. The first collection was performed preoperatively before biotalcum application, and then during the time of losses from thoracic drainage bigger than 150 ml in 24 hours. The duration of thoracic drainage was 4 days +/- 1 day. The success of the treatment was observed by ultrasound scan before drainage removal; during the first three months always at intervals of 1 month and then after 3 and 6 months, depending on the progression of a disease. RESULTS: No reccurence occured when the P-CRP (pleural) and S-CRP (serum) ratio exceeded 60% during the first 48 hours after pleurodesis. On the other hand, when the ratio fall bellow 30-35%, the effusion relapsed frequently. CONCLUSION: The P-CRP/S-CRP ratio as a promising marker of talc pleurodesis effectiveness and monitoring both P-CRP and S-CRP levels is inexpensive and acceptable method for clinical practice. The pleural effusion caused by malignant mesothelioma appeares to be resistant to talc pleurodesis.

[Cardiac surgery operations and their influence on serum level of antiinflammatory cytokine interleukin-10]. / Kardiochirurgické operace a jejich vliv na sérovou hladinu protizánetového cytokinu interleukinu-10.

BACKGROUND: Cardiac surgical operation is followed by the development of inflammatory reaction. This reaction is regulated in many ways including the production of antiinflammatory cytokines such as IL-10 to avoid potentially harmful effects of inflammation. METHODS AND RESULTS: We compared serum levels of cytokines IL-10, IL-6, and IL-13 in the group of patients undergoing cardiac surgical operation using either cardiopulmonary bypass (CPB, n=17) or surged on the beating heart (n=17). We found significant elevation in the serum level of IL-10 during surgery with the peak immediately after finishing surgery in CPB patients and at the first postoperative day in non-CPB patients, respectively. There is statistically significantly higher level of IL-10 in CPB patients in comparison with non-CPB patients at the end of surgery. Serum level of IL-6 is elevated in both groups during surgery reaching maximum immediately after surgery in CPB patients and at the first postoperative day in patients without CPB, respectively. The serum levels of IL-13 are only nonsignificantly changed during operation and in postoperative period in both groups. CONCLUSIONS: The intensity of inflammatory response in CPB patients which is enhanced by massive contact activation of blood and extensive ischemia-reperfusion injury is regulated by the production of antiiflammatory IL- 10 cytokine.

[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.

[Cytoimmunofluorometry in the diagnosis of MALT lymphoma of the gastrointestinal tract]. / Cytoimunofluorometrie v diagnostice MALT lymfomu gastrointestinálního traktu.

MALT lymphoma is raised from lymphoid cells of mucosa associated lymphoid tissues in a long-term, multistep process of the accumulation of genomic damage very often under the influence of chronic (e.g. Helicobacter pylori) infection. Cytoimmunofluorometric analysis of cell suspensions obtained from mechanically disintegrated bioptic samples of gut mucosa is a very useful approach to detect the presence of MALT lymphoma cells. MALT lymphoma cells are in a great majority of B cell origin. Malignant cells are characterized by the typical characteristic SS parameter, strong expression of CD20 and CD79b molecules, and by the absence of CD5 and CD23 molecules. The ultimate evidence of clonality is immunoglobulin light chains either kappa or lambda expression determination.

[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.