Biomedical subjects

J Cerný

Publications and source records attributed to J Cerný.

At least 19 recordsLinked to original sources

PubMed2006

[Massive pulmonary embolism--attempt at embolectomy following the failure of thrombolytic treatment].

We report a case of a 61-year-old woman with progressive dyspnoea, fatigue and tiredness over the last two weeks. Based on physical examination on admission, ECHO and CT exams, a diagnosis of massive pulmonary embolism was confirmed and the patient received thrombolytic treatment. Since no response to thrombolysis was apparent and the patient was clinically unstable with persisting signs of right ventricular dysfunction in echocardiography, a rescue surgical embolectomy was ordered and performed using cardiopulmonary bypass. Multiple large clots were removed from both pulmonary arteries. Following embolectomy, severe irreversible right ventricular failure developed and the patient died. Autopsy revealed large blood clots in peripheral pulmonary circulation causing terminal right-heart failure. We discuss the therapeutic options in massive pulmonary embolism and the potential role of rescue surgical embolectomy.

Embolectomy↗

PubMed2005

Blood phagocyte activation during open heart surgery with cardiopulmonary bypass.

Open heart surgery with a cardiopulmonary bypass (CPB) is associated with a systemic inflammatory response which significantly contributes to adverse postoperative complications. The purpose of this study was to characterize the activation of blood phagocytes during open heart surgery with CPB. Blood samples were collected during and up to 24 h after surgery. The production of reactive oxygen species (ROS) in whole blood, the expression of surface molecules by blood phagocytes and complement activity in the plasma were determined. A cDNA microarray analysis of leukocyte RNA profile of genes was performed related to the inflammatory response. Activation of the complement was already observed at the beginning of CPB. This was followed by an increase in the neutrophil number and in both spontaneous and opsonized zymosan-activated ROS production after the onset of reperfusion. The activation of blood phagocytes was affirmed by changes in surface receptors involved in the adhesion and migration of leukocytes (CD11b, CD62L and CD31). Gene arrays also confirmed the activation of leukocytes 4 h after reperfusion. In conclusion, open heart surgery with a cardiopulmonary bypass was found to be associated with a rapid and pronounced activation of blood phagocytes and complement activation which was partly independent at the onset of CPB.

Aged↗

PubMed2005

The use of comparative duplex PCR in monitoring of patients with non-Hodgkin's lymphoma and chronic lymphocytic leukaemia.

Various quantitative PCR approaches have been utilized during the last years to provide information about the treatment efficacy and the risk of recurrent disease in haematological malignancies. Apart from the frequently used real-time PCR, cost-saving modified standard PCR methods may be applied as well. This report evaluates the utility of the end-point comparative duplex PCR. We have used this method for monitoring of 35 patients with either NHL or CLL and observed a good correlation between quantitative molecular results and clinical outcome. There was also an agreement between comparative duplex PCR and real-time PCR in patients who were monitored by both methods. We therefore believe that use of this technique should be strongly considered instead of simple qualitative detection in monitoring of therapeutic outcome in NHL or CLL patients.

Genetic Markers↗

PubMed2003

[Molecular diagnosis of infections].

Molecular diagnostics (detection of nucleic acids by molecular genetics techniques) become more valuable in clinical diagnosis of disease. Apart from the already long-time used genetic techniques for detection of congenital anomalies, current use of molecular techniques includes detection of microbial pathogens. The character of these techniques increases the possibility of achieving diagnosis in cases where classical cultivation is not possible, is not reliable or is not fast enough. As with every new approach, molecular diagnostics have faced encountered reactions from the scientific community. Some scientists tend to overestimate the value of molecular diagnostic techniques, while sceptics, sometimes influenced by a biased or incomplete knowledge of the technology, think it is of little value. In this work, on the basis of literature and our own data from more than 5 years of experience with these methods, we have assessed the pros and cons of the use of molecular diagnostics of infectious diseases in the light of their potential use in clinical practice.

Bacterial Infections↗

PubMed2003

[The results of kidney transplantation from very young paediatric cadaveric donors to adult recipients in Czech Republic in the years 1994-2001 (retrospective study of the Czech transplant centres)].

The results of kidney transplantation from very young paediatric cadaveric donors up to five years, which were transplanted to adult recipients, are evaluated in the first retrospective study of all Czech transplantcentres. In general, 42 of these transplantations were carried out during 1994-2001. In 28 cases single kidney was transplanted, in 14 cases en bloc graft of both children kidneys was transplanted. The reasons of kidney failure by recipients are usual (in 42.9% glomerulonephritis). An average age of the donors was 34.7 months (median 39 months). An average age of the recipients during transplantation was 42.6 years (median 43.5 years). All the kidneys were placed into retroperitoneum. As long as only single kidney was transplanted, the rules, already propagated by Salvatiera in 1970, were respected. At the transplantation of en bloc graft, Kinne's method with possible vessels elongation of the graft and with uretero-uretero anastomoses (in so called Ostrava modification) was used. The higher occurrence of the primary graft non-function (as 50% losses of all grafts) was confirmed in the study. Its reason was an acute thrombosis of vessels or rotation of graft stem. One-, three- and five-years patients and grafts survival were 97.6-90.5-90.5% and 76.2-73.8-73.8% actually. Patients survival are very good while grafts survival are average, momentarily a bit worse than national data published by the Czech Transplant Society, which determines one-, three- and five-years grafts survival with 90.3-82.9-75.5% and 79.1-71.1-65% actually. The kidney transplantation from paediatric cadaveric donors to adult recipients is acceptable under present allocated criteria, in case that some principles are kept. The study confirms the international experiences about rather higher incidence of surgical complications and primary graft non-function. In conclusion, some general references for reduction of complications, as well as for improvement of these transplantation results are introduced.

Adult↗

PubMed2002

[Clinical importance of semi-quantitative monitoring of lymphomas using the comparative polymerase chain reaction].

BACKGROUND: PCR techniques detecting interchromosomal translocation and clonal immunoglobulin gene rearrangement (IgH) as disease markers in non-Hodgkin's lymphomas (NHL) has been utilised past ten years. However, qualitative PCR detection of persisted minimal residual disease cannot provide clinically useful prognostic information and presently, quantitative approaches are required to predict patient outcome and assess response to the treatment. In some cases, "end-point" quantifying techniques, such as comparative PCR, are applicable and the relative estimation of differences in target quantity may serve in disease monitoring rather than absolute number of target copies. METHODS AND RESULTS: Our method of comparative PCR employs co-amplification of sequences of interest (clonal CDR3, bcl2/Jh) and the segment of Hras 1 gene(ras) as an internal standard. Serial dilutions of stored diagnostic DNAs from blood and bone marrow are examined in the same PCR and, after gel densitometry, the amount of initial target is assessed by comparing exponential products of co-amplification. The comparative PCR assay was utilized in monitoring of NHL patients cured either with conventional therapy, or with high-dose regimens and transplantation with stem cells, or with chimaeric anti-CD20 monoclonal antibody (Rituximab). Results from 50 monitored intervals obtained during several months up to several years were supplemented with clinical statements retrospectively. Some of patients became PCR-negative, reappearance of PCR-positivity was observed as well. The decrease or increase of disease marker corresponded to clinical observations. Results obtained from bone marrow were in agreement with those obtained from blood. CONCLUSIONS: End-point quantifying PCR comparative assay may provide an information on the increased risk of relapse and impact of the therapy. The predictive value of these methods depends on the frequency of sample taking and on the sensitivity of the method, which should be monitored in negative cases.

Complementarity Determining Regions↗

PubMed2002

Pericardial constriction caused by Candida albicans.

An uncommon occurrence of constrictive pericarditis caused by Candida albicans and its treatment by successful pericardectomy and epicardectomy are described. For pathogen detection, both cultivation and molecular diagnostics were used. The speed and reliability of molecular diagnostics using polymerase chain reaction make this method a powerful tool for pathogen detection in any clinical specimen.

Aged↗

PubMed2002

Binding of the Galanthus nivalis agglutinin to thymocytes reveals alterations in surface glycosylation during T-cell development.

Surface binding of the Galanthus nivalis agglutinin (GNA) to thymocyte subsets has been studied in pigs and rodents by multicolour flow cytometry. In all the species examined, analogous staining profiles have been recorded. Counter-staining with anti-CD3epsilon, anti-CD4 and anti-CD8 monoclonal antibodies (MoAb) revealed that a significant increase of the GNA targets on the cell surface occurred during early thymocyte differentiation and reached its maximum at the level of the CD3loCD4+CD8+ small cortical thymocyte. This was followed by a decrease in the GNA binding capacity upon terminal maturation to the single positive thymocytes. PAGE analysis has revealed a dominant GNA-binding glycoprotein (molar mass approx. 90 kDa) present on thymocyte plasma membranes and absent on the surface of splenic lymphocytes, although both the whole cell lysates from both organs contained GNA ligands of the same size. Our findings are in agreement with previous data showing that immature thymocytes differ from their mature counterparts and peripheral T lymphocytes in the surface glycosylation pattern, and support the hypothesis that lectin-glycoprotein interaction plays a significant role in the cell-to-cell crosstalk in the thymic cortex.

Animals↗

PubMed2002

[TT virus infection in liver transplant recipients with cryptogennic cirrhosis].

The possible causative role of novel TT virus in liver diseases has been intensively studied in regarding its hepatotrophy, ability to cause persistent infection and worldwide prevalence. The aim of this study was to estimate the prevalence as well as the clinical importance of TTV in a normal healthy population group in the Czech Republic and in a group of liver transplant recipients diagnosed with cryptogenic cirrhosis. Polymerase chain reaction (PCR) detected the DNA of TT virus in 68% (13/19) of samples isolated from peripheral blood leukocytes and in 21% (4/19) of plasma samples in the liver transplant group. The viral DNA was detected only in 11.8% (4/34) of leukocytes and in no plasma sample from the healthy population control group. All patients included in this study had good liver function and had no complications during the postoperative period. The prevalence of TTV DNA detection in healthy control group in Czech republic is similar to the rates reported in European and North American countries. Significant difference was proved between the prevalences of TTV in the groups of healthy controls and liver transplant recipients with cryptogenic cirrhosis. However, no association of TTV infection with possible postoperative complications could be found.

Adult↗

PubMed2002

Influence of polysulfone and hemophan hemodialysis membranes on phagocytes.

The aim of the study was to compare the effect of hemophane and polysulfone membranes on the phagocyte-derived production of reactive oxygen species (ROS) as well as on neutrophil CD11b and CD62L expression in patients undergoing regular hemodialysis. The effects of hemodialysis membranes were also studied in in vitro conditions after coincubating them with differentiated HL-60 cells. ROS production was measured using chemiluminometric and flow cytometric methods. Expression of CD11b, CD62L and mitochondrial membrane potential were detected by monoclonal antibodies and by the JC-1 fluorescent probe, respectively. Depressed ROS production was observed in patients already before dialysis. Further decrease in ROS production and an increase in CD11b expression were observed especially in patients after hemophan hemodialysis. Decreased ROS production and increased CD11b expression were observed also after incubation of HL-60 cells with hemophan membranes. Mitochondrial membrane potential dropped only after incubating cells with hemophan membranes proving its more serious adverse effects in comparison with the polysulfone membrane. In conclusion, deleterious effects of hemodialysis on the metabolic activity of phagocytes were proved. Combining chemiluminescent and flow cytometric methods for the detection of ROS production and determining mitochondrial membrane potential can be useful tools for the analysis of material biocompatibility.

Aged↗

PubMed2002

[Will ECG combined with echocardiography detect viable myocardium in patients after myocardial infarct?].

The objective of the submitted trial was to use combined examination of ECG at rest and after exercise to detect a viable heart muscle in patients with a postinfarction Q wave and ST elevations. The authors found that according to the character of changes of the ST elevation in ECG after exercise it is not possible to detect patients with a viable myocardium. During the combined procedure using analysis of the kinetic disorder at rest and increase of the ST segment in ECG after exercise it is possible in patients with hypokinesia or akinesia at the site of the Q wave to differentiate successfully 66% patients with a viable heart muscle. This combined procedure can serve as a selective test for implementation of further more accurate diagnostic tests for the detection of a viable heart muscle because in common clinical practice the above data are available in these patients before indication of more complicated examinations.

Coronary Artery Bypass↗

PubMed2001

Peri- and post-operative course of cytokines and the metabolic activity of neutrophils in human liver transplantation.

Peri- and post-operative (up to day 7 after surgery) neutrophil chemiluminescence and the plasma concentrations of interleukin 6 (IL-6), IL-8, IL-10 and tumour necrosis factor alpha (TNF-alpha) were evaluated in the blood of patients undergoing liver transplantation. IL-6, IL-8 and IL-10 levels increased during early reperfusion and then returned to normal mostly within the first post-operative day. TNF-alpha was increased during the whole period observed. Spontaneous as well as activated neutrophil chemiluminescence was depressed in early reperfusion and remained low during the whole period followed. Samples collected during early reperfusion provided positive correlation for IL-6 vs IL-8 as well as for IL-6 and IL-8 vs chemiluminescence. The data were also evaluated with respect to the outcome of transplantation. Since IL-8, IL-10 and TNF-alpha levels increased significantly during the first post-operative week, mainly in a group of patients who developed serious complications within the first month after surgery, we proved a connection between peri- and early post-operative induction of cytokine release and the outcome of liver allograft transplantation.

Adult↗

PubMed2001

A proteolytically truncated form of free CD18, the common chain of leukocyte integrins, as a novel marker of activated myeloid cells.

An unusual CD18 monoclonal antibody (mAb) MEM-148 binds, in contrast to standard CD18 mAbs, specifically to peripheral blood monocytes and neutrophils activated by various stimuli such as phorbol myristate acetate, opsonized zymosan, heat-aggregated immunoglobulin, and (after priming with lipopolysaccharide, tumor necrosis factor, or granulocyte-macrophage colony-stimulating factor) also by formyl-methionyl-leucyl-phenylalanine. In addition, in vivo activated neutrophils obtained from urine of patients following recent prostatectomy were also strongly positive for MEM-148. On the activated myeloid cells the mAb recognized a 65- to 70-kd protein identified immunochemically and by mass spectrometric peptide sequencing as a membrane-anchored fragment of CD18 (the common chain of leukocyte integrins) produced by proteolytic cleavage. The CD18 fragment originated mainly from integrin molecules stored intracellularly in resting cells, it was unassociated with CD11 chains, and its formation was inhibited by several types of protease inhibitors. Thus, the 65- to 70-kd CD18 fragment represents a novel abundant activation marker of myeloid cells of so far unknown function but possibly involved in conformational changes in leukocyte integrin molecules resulting in increased affinity to their ligands.

Antibodies, Monoclonal↗

PubMed2001

A novel anti-CD18 mAb recognizes an activation-related epitope and induces a high-affinity conformation in leukocyte integrins.

Monoclonal antibody MEM-148 was previously shown to recognize CD18 chains in a free form unassociated within leukocyte integrin heterodimers, but yet it is paradoxically able to induce a high-affinity conformation in the native, cell surface expressed LFA-1 molecules. Our results based on kinetics of binding, immunoprecipitation and cell-aggregation experiments demonstrate that the mAb does bind to and stabilizes a specific conformation of LFA-1 heterodimers apparently distinguished by an increased affinity to its cellular ligand(s). A similar high-affinity conformation of LFA-1, in which the MEM-148 epitope becomes exposed, is induced also by a Mg2+/EDTA or low pH (5.5-6.5) treatments which may mimic physiologically relevant situations in normal or inflamed tissues. Thus, mAb MEM-148 is a novel valuable tool for detection and induction of specific conformations of human leukocyte integrins.

Animals↗

PubMed2001

Biliary complications in liver transplantation.

OBJECTIVES: Biliary complications (BC) continue to be a major cause of morbidity among liver transplant recipients. The aim of this study was to analyse the incidence, risk factors and management of biliary tract complications at the Centre for Cardiovascular Surgery and Transplantations in Brno. METHODS: Between January 1992 and December 2000, 118 orthotopic LT were performed in 113 patients. Reconstructions of biliary tract included four methods: end-to-end choledochocholedochostomy with a drain in the retained gallbladder in 15 cases, end-to-end or side-to-side choledochocholedochostomy with a T tube in 29 cases, end-to-end choledochocholedochostomy without a T tube in 67 cases and choledochojejunostomy over the drain in 7 cases. Biliary complications were divided into three groups: stenosis, leaks and cholangitis. RESULTS: Biliary complications occurred after 33 LT (27.9%), affecting 28.3% of the recipients. They occurred most frequently when CCwT or CJ types of reconstruction were used (44.8% and 57.1%, respectively) and least frequently in the CCw/oT group of patients (16.4%). The most common type of biliary complication was stenosis which occurred in 20 patients (in 12 at the anastomotic site, in 5 it was caused by external compression and 3 had nonanastomotic strictures); biliary leaks were in 13 patients and cholangitis in 10 patients. Twenty three biliary complications occurred in the early postoperative period and 12 during the follow-up. Endoscopic treatment was used as primary therapy in 17 patients, primary surgical intervention was used in 12 patients and five patients were treated conservatively. Five patients died due to biliary complications (mortality, 15.1%). CONCLUSIONS: It can be concluded that BC is a common cause of morbidity after LT. In the last few years, its frequency has remained constant. Technical failure or local ischemia are major causes. Biliary leaks predominate in the early posttransplant period. Since end-to-end choledochocholedochostomy without a T tube is associated with the lowest incidence of BC, it is considered to be the method of choice. BC can usually be managed endoscopically, although early leaks frequently require reoperation. Aggressive and early management of BC can reduce mortality in patients after LT.

Adolescent↗

PubMed2000

Human leukocytes contain a large pool of free forms of CD18.

Monoclonal antibodies to CD18, the common chain of leukocyte integrins, recognize in various types of human lymphoid and myeloid cells under the conditions of nonreducing Western blotting three species of CD18 of mol. wt. 96, 87, and 78 kDa, respectively. Using a unique monoclonal antibody MEM-148 reacts exclusively with free CD18 molecules, but not with leukocyte integrin heterodimers. We demonstrate that only the upper one (96 kDa) is present on the cell surface within the CD11/CD18 integrin heterodimers, while the lower ones (87 and 78 kDa) are found intracellularly as free molecules unassociated with CD11 chains or other molecules. These intracellular free CD18 chains may in part represent biosynthetic precursors; alternatively, these species may represent an intracellular source of the recently observed free, proteolytically truncated CD18 chains expressed on the surface of activated myeloid cells.

Antibodies, Monoclonal↗

PubMed2000

Phenotypic effects of CD3zeta targeting into glycosphingolipid-enriched membrane microdomains (GEMs) of T cells.

In the present study we tested whether the forced expression of the CD3zeta chain within detergent-resistant, glycosphingolipid-enriched membrane microdomains (GEMs) will result in a constitutively activated phenotype in human T cells. To this aim, a monomeric recombinant protein (LckSH4-CD3zeta), containing the intracellular part of human CD3zeta chain fused to N-terminal double-acylation motif (SH4 domain) of protein tyrosine kinase Lck, was expressed in Jurkat human T lymphoid cell line and its Lck-negative mutant, J. CaM1.6. The Lck SH4 domain indeed predominantly targeted the chimeric protein into GEMs. In transfectants derived from wild-type Jurkat cells, but not in those derived from the Lck-deficient mutant, the LckSH4-CD3zeta protein was constitutively tyrosine-phosphorylated. Tyrosine phosphorylation of a major Jurkat cell phosphoprotein (pp85) was diminished in the transfectants. However, the transfectants did not exhibit any features of constitutively activated T cells, and their responses to anti-CD3 treatment were very similar to the wild-type Jurkat cells. Thus, the constitutive expression of this form of CD3zeta chain in GEMs is not sufficient for eliciting an activated state in the Jurkat cells.

CD3 Complex↗

PubMed2000

Phosphoprotein associated with glycosphingolipid-enriched microdomains (PAG), a novel ubiquitously expressed transmembrane adaptor protein, binds the protein tyrosine kinase csk and is involved in regulation of T cell activation.

According to a recently proposed hypothesis, initiation of signal transduction via immunoreceptors depends on interactions of the engaged immunoreceptor with glycosphingolipid-enriched membrane microdomains (GEMs). In this study, we describe a novel GEM-associated transmembrane adaptor protein, termed phosphoprotein associated with GEMs (PAG). PAG comprises a short extracellular domain of 16 amino acids and a 397-amino acid cytoplasmic tail containing ten tyrosine residues that are likely phosphorylated by Src family kinases. In lymphoid cell lines and in resting peripheral blood alpha/beta T cells, PAG is expressed as a constitutively tyrosine-phosphorylated protein and binds the major negative regulator of Src kinases, the tyrosine kinase Csk. After activation of peripheral blood alpha/beta T cells, PAG becomes rapidly dephosphorylated and dissociates from Csk. Expression of PAG in COS cells results in recruitment of endogenous Csk, altered Src kinase activity, and impaired phosphorylation of Src-specific substrates. Moreover, overexpression of PAG in Jurkat cells downregulates T cell receptor-mediated activation of the transcription factor nuclear factor of activated T cells. These findings collectively suggest that in the absence of external stimuli, the PAG-Csk complex transmits negative regulatory signals and thus may help to keep resting T cells in a quiescent state.

Adaptor Proteins, Signal Transducing↗