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Biomedical subjects

J Polák

Publications and source records attributed to J Polák.

At least 19 recordsLinked to original sources

[The use of quantitative assessment of Wilms tumour gene 1 for monitoring of residual disease in acute myeloid leukemia patients].

BACKGROUND: Despite a considerable effort, the majority of acute myeloid leukaemia (AML) patients do not have a suitable specific molecular marker for monitoring minimal residual disease (MRD). The results of some studies suggest the Wilms tumour gene (WT1) as a possible molecular marker of MRD. METHODS AND RESULTS: We measured the expression of WT1 at diagnosis and during treatment of the acute myeloid leukaemia (AML) patients. The expression of WT1 was measured by the quantitative real-time RT-PCR in peripheral leukocytes from 56 AML at diagnosis and 7 patients with AML transformed from myelodysplastic syndromes (MDS). The WT1 expression was significantly elevated (up to 3 orders of magnitude) in peripheral blood samples (PB) of AML patients at diagnosis compared to PB samples of healthy donors (P < 0.0001). The level of WT1 expression depends particularly on FAB AML subtype, with the highest being found in AML patients with subtypes M4, M1, M3 and AML transformed from MDS. Conversely, AML patients with M2 and with the presence of AML1/ET0 at presentation showed a significantly lower expression of the WT1 gene compared to the remaining AML patients at presentation (P = 0,005). Further, sequence samples of 12 AML patients under long-term surveillance were tested for the WT1 expression in parallel with the expression of specific MRD markers--fusion genes: AMLI/ETO, PML/RARalpha and CBFB/MYH11. The levels of WT1 gene expression and the above specific fusion genes significantly correlated. Moreover, 14 patients without the specific MRD marker were tested for the WT1 expression. The results show that haematological relapses were associated with the rise of expression of the specific fusion genes and with the WT1 gene expression. The rise of WT1 expression above the level seen in leucocytes from peripheral blood and/or bone marrow of healthy donors--in four patients under long-term surveillance the "molecular relapse" predicted ongoing haematological relapses as early as 2 months in advance. CONCLUSIONS: Our results, in accordance with some of the previously published ones, show that WT1 expression seems to be a suitable marker of minimal residual disease in AML patients.

Acute Disease↗

[Effect of dynamic strength training on insulin sensitivity in men with insulin resistance].

BACKGROUND: Physical activity is generally accepted as a part of the nonpharmacological therapy of the insulin resistance. Endurance training is generally recommended as an appropriate approach. The aim of this study was to assess the effect of three-month dynamic strength training on the insulin sensitivity in middle-aged men with insulin resistance. METHODS AND RESULTS: 10 men (5 obese non diabetics and 5 overweight patients with diabetes mellitus type 2 (age 51.36+/-7.25 years, average weight 110.16+/-13.56 kg and BMI 33.22+/-3.52 kg/m2 underwent a three-month dynamic strength training at the level of 60 to 70 % of their maximum muscle strength (one-repetition maximum 1-RM). Insulin sensitivity was determined using the hyperinsulinic euglycemic clamp before and after the training period. Training promoted to increase the muscle strength (p<0.001). It did not induce changes in body weight, body composition and maximum aerobic capacity. The training induced an increase in insulin sensitivity (glucose disposal M: 3.0 vs 4.0 M - mg/min/kg, p<0,01). CONCLUSIONS: Dynamic strength training improves insulin sensitivity in men with insulin resistance independently on weight loss or increase in aerobic capacity. Our results suggest that the dynamic strength training is an appropriate physical activity for management of the insulin resistance.

Diabetes Mellitus, Type 2↗

[Expression of cyclin-dependent kinase inhibitors in leukemia].

BACKGROUND: Leukemias develop due to defects in proliferation, differentiation and apoptosis, which take place in stem cells or progenitors of hematopoiesis. These processes have several crossing points, one of them is the role of inhibitors of cyclin-dependent kinases. The aim of this study was to study the expression of cyclin-dependent kinases inhibitors p21 Cip and p27 Kip and expression of proliferative antigen Ki-67 in leukocytes of human leukemia. METHODS AND RESULTS: The expression of cyclin-dependent kinases inhibitors was detected at mRNA level mainly by comparative reverse-transcription polymerase chain reaction and in selected samples also by the real-time polymerase chain reaction. While p27 Kip expression in leukocytes of leukemic patients and healthy persons was universal, large differences in expression of p21 Cip were found both among individual patients of the same type of leukemia and between different types of leukemias and healthy persons. The p21 Cip expression was significantly higher in acute leukemias than in chronic ones and healthy persons. A comparison of p21 Cip expression with the clinical outcome of the leukemic patients showed that the group of 14 acute leukemia patients surviving more than 30 months had a significantly lower expression of p21 Cip than 12 patients of this type of leukemia who died within this time limit. Moreover, the results obtained on a smaller set of acute promyelocytic leukemia patients indicated that the lower p21 expression is connected with a better prognosis. CONCLUSION: Our results pointed out the importance of the cyclin-dependent kinase inhibitor p21 Cip in human leukemias and indicated that the lower p21 Cip expression might be a positive prognostic factor in acute myeloid leukemia patients.

Cell Cycle Proteins↗

Iron deprivation induces apoptosis independently of p53 in human and murine tumour cells.

Iron deprivation induces apoptosis in some sensitive cultured tumour cells, while other cells are resistant. In order to elucidate the mechanisms involved in apoptosis induction by iron deprivation, we studied the expression of p53 and the expression of selected p53-regulated genes. To discriminate between changes coupled only with iron deprivation and changes involved in apoptosis induction by iron deprivation, we compared the expression of the genes in sensitive (human Raji, mouse 38C13) versus resistant (human HeLa, mouse EL4) cells under iron deprivation. Iron deprivation was achieved by incubation in a defined iron-free medium. The level of p53 mRNA decreased significantly under iron deprivation in sensitive cells, but it did not change in resistant cells. On the contrary, the level of the p53 protein under iron deprivation was slightly increased in sensitive cells while it was not changed in resistant cells. The activity of p53 was assessed by the expression of selected p53-regulated targets, i.e. p21(WAF1/CIP1) gene, mdm2, bcl-2 and bax. We did not detect any relevant change in mRNA levels as well as in protein levels of these genes under iron deprivation with the exception of p21(WAF1/CIP1). We detected a significant increase in the level of p21 mRNA in both (sensitive and resistant) mouse cell lines tested, however, we did not find any change in both (sensitive and resistant) human cell lines. Moreover, the p21(WAF1/CIP1) protein was accumulated in mouse-sensitive 38C13 cells under iron deprivation while all other cell lines tested, including human-sensitive cell line Raji, did not show any accumulation of p21(WAF1/CIP1) protein. It seems that the p21(WAF1/CIP1) mRNA, as well as protein accumulation, is not specifically coupled with apoptosis induction by iron deprivation and that it is rather cell-line specific. Taken together, we suggest that iron deprivation induces apoptosis at least in some cell types independently of the p53 pathway.

Animals↗

[Lymphangioleiomyomatosis in a menopausal woman].

Lymphagioleiomyomatosis is a rare pulmonary disease which affects mainly women of childbearing age. An observation of lymphangiomyomatosis in a menopausal woman or in men is considered very unusual. We report a case of lymphagioleiomyomatosis in a 60-year-old woman.

Female↗

Location of the BCR/ABL fusion genes on both chromosomes 9q34 in Ph negative chronic myeloid leukemia.

We present two patients with Ph-negative chronic myeloid leukemia (CML) and fusion signal BCR/ABL on both chromosomes 9, located in region 9q34. The first case was a 27 years old man with CML. Molecular studies (RT-PCR) revealed the rearrangement in the major-BCR region and expression of chimeric BCR/ABL mRNA of b3a2 configuration. By classical cytogenetic studies (G-banding) karyotype 46,XY was found in short-term cultivated bone marrow cells and phytohemagglutinin (PHA) stimulated peripheral lymphocytes. FISH studies revealed the BCR/ABL fusion signals on both chromosomes 9 and green BCR signals on both chromosomes 22 in all mitoses studied. Detection of the alleles of ABL1 intragenic STR locus by fluorescence PCR followed by fragmentation analysis in the patient and his parents provided no information about transmission of the ABL gene. Quantitative assessment of BCR/ABL transcript level by RT-PCR showed 60 and 70% BCR/ABL positivity in two peripheral blood samples at 6,5 and 10,5 months after diagnosis, respectively, which does not correspond to the expression from two identical BCR/ABL hybrid genes. Therefore, the possible mechanism of the origin of two BCR/ABL fusion signals present on both chromosomes 9 could not be resolved and remains speculative. The second case was a 53 years old male with diagnosis of chronic phase of CML, with first sign of acceleration one month after diagnosis and death because of sepsis in blastic phase within four months. The cytogenetic findings were identical to those in case No. 1., i.e. karyotype 46, XY by G-banding, two BCR/ABL fusion signals on both chromosomes 9 and RT-PCR molecular studies proved b3a2 breakpoints. It is generally accepted that prognosis of the patients with fused BCR/ABL gene located on chromosome 9 is poor. The presence of two fused genes could be anticipated as two Ph chromosomes in accelerated and blastic phases of the disease. However, in our study, quantitative findings of BCR/ABL transcripts did not corresponded to the expression of two BCR/ABL genes originating from duplication. If this assumption is correct then the expression of both fused genes BCR/ABL was in case No. 1 equally suppressed and total expression reached about the level of one BCR/ABL gene.

Adult↗

P230 BCR/ABL protein may be associated with an acute leukaemia phenotype.

The BCR/ABL rearrangement, the molecular hallmark of chronic myelogenous leukaemia (CML), is rare in acute myeloid leukaemia (AML), being detected in approximately 1% of cases. In the vast majority of CML cases the breakpoint on chromosome 22 falls in the so-called major breakpoint cluster region of the BCR gene. Only a few cases of CML with breakpoint in the minor or in the micro bcr region have so far been reported. The micro breakpoint position has been associated mainly with a mild form of CML, defined as Philadelphia chromosome-positive chronic neutrophilic leukaemia (Ph-positive CNL). Using reverse transcription-polymerase chain reaction (RT-PCR) we report a patient with an acute myeloid leukaemia phenotype at diagnosis who showed a BCR/ABL rearrangement with a breakpoint located in the micro bcr region (e19a2 junction). Cytogenetic analysis showed a progression of the malignant clone, finally leading to cells with two Ph chromosomes, trisomy 8, isochromosome 17q and deletion of the long arms of chromosome 7. The findings of chromosomal changes point to a possibility of blast crisis of CML with a clinically silent chronic phase. Immunoprecipitation and auto-phosphorylation assay revealed the expression, by the patient's blast cells, of an abnormal P230 BCR/ABL protein, which showed for the first time that this protein was constitutively activated in primary cells from patients. This finding may contribute to the understanding of the role of the BCR/ABL rearrangements in determining different leukaemia phenotypes ranging from acute lymphoid and myeloid leukaemias to mild chronic neutrophilic leukaemias.

Acute Disease↗

Relative concentration of plum pox virus in leaves and flowers of some Prunus species and cultivars.

The relative concentration of plum pox virus (PPV) in leaves and flowers of plum, damson, myrobalan, blackthorn, apricot and peach trees was determined by enzyme-linked immunosorbent assay (ELISA) and expressed as the lowest dilution with positive reaction. Significant differences in relative PPV concentration were found in leaves among individual Prunus species naturally or artificially infected with the virus. The highest relative PPV concentration was found in blackthorns (7.81 x 10(-4)), common plum and apricot (1.56 x 10(-3) for the both latters). Wild growing PPV-infected plums and blackthorns can be considered equally important source of sharka infection as PPV-susceptible cultivars of plums, apricots and peaches. High PPV concentration in flowers is of diagnostical value. High variability of relative PPV concentration was observed inside the species among individual cultivars. Susceptible cultivars were characteristic by high relative PPV concentration, e.g. apricot cvs. Vegama (9.8 x 10(-5)) and Velkopavlovická (1.95 x 10(-4)), and peach cvs. Maria Emilia (7.81 x 10(-4)) and Harbinger (1.56 x 10(-3)). On the other hand, cultivars resistant to PPV were characteristic by very low relative PPV concentration, e.g. apricot cv. Stark Early Orange (5 x 10(-2)) and peach cvs. Envoy (5 x 10(-2)) and Favorita Morettini (2.5 x 10(-2)). The highest relative PPV concentration was found in young trees newly infected with PPV.

Enzyme-Linked Immunosorbent Assay↗

[High-resolution computer tomography in the diagnosis of cryptogenic fibrosing alveolitis. Correlation with pulmonary function and bronchoalveolar lavage].

BACKGROUND: High resolution computed tomography (HRCT) revealed new findings as regards changes associated with cryptogenic fibrotizing alveolitis. So far views on changes suggesting activity of the disease are not uniform and they were not yet confronted with examinations of fluid obtained by bronchoalveolar lavage (BALT). The objective of the present work is to evaluate the contribution of HRCT to the assessment of the activity of the disease and compare the findings with the examination of BALT. METHODS AND RESULTS: The authors evaluated the HRCT examination made in 52 patients with histologically confirmed cryptogenic fibrotizing alveolitis and in 39 of them the finding was confronted with the BALT examination. Signs of active inflammatory disease according to HRCT comprised a ground-glass increase of the density in 80.8%, centrilobular micronodules in 17.3% and a prominence of centrilobular structures in 32.7%. The suggested extent of affection of the pulmonary parenchyma assessed according to HRCT correlated well with the degree of dyspnoea, restrictive and obstructive ventilation disorder, pulmonary diffusion and with hypoxaemia. The authors confirmed also an inverse correlation between the grade assessed by HRCT and the percentage of lymphocytes in BALT. CONCLUSIONS: The investigation confirmed the contribution of HRCT in the diagnosis of patients with cryptogenic fibrotizing alveolitis, it confirmed the complementary importance of HRCT and BALT examination when assessing the activity of the disease and its asset when monitoring patients during treatment.

Adolescent↗

Evaluating lead bioavailability data by means of a physiologically based lead kinetic model.

A method of bioavailability estimation is presented in which a physiologically based kinetic model of lead kinetics is fit simultaneously to blood and bone lead concentrations after a period of exposure to dietary lead. Optimization of the simultaneous fit, varying only fractional absorption, gives the best estimate of fractional bioavailability for each treatment group. The analysis was applied to data from three separate studies in which rats were fed for 30 consecutive days purified diets containing lead added as lead acetate, mine waste-contaminated test soils, or mine waste itself. Fractional absorption decreased as lead intake increased, regardless of the source of the lead; but the magnitude of this dose dependence was lead source-dependent. There were no differences in lead absorption by male and female rats when lead intake was expressed per unit body weight. Fractional absorption varied from 4 to 5%, at low exposure rates (1-2 mg lead/kg/day) when lead acetate was added to the diet, to 0.24% at a high exposure rate (24 mg/kg/day) when a mine waste-contaminated test soil was added to the diet. Comparison of the results of this analysis with the results of a more conventional analysis, in which the bone and blood lead concentrations were separately compared with bone and blood lead concentrations in rats given daily injections of lead acetate intravenously for 29 consecutive days, demonstrated that the standard analysis failed to reveal the dose dependence of fractional absorption.

Animal Feed↗

Multiple forms of cathepsin B in human lung cancer.

In this study we have examined, by means of isoelectric focusing (IEF) in native polyacrylamide gel and contact-print fluorescence zymography, whether human lung carcinomas and the lung parenchyma contain different pools of multiple charge forms of the cysteine proteinase cathepsin B. The isoelectric point (pI) patterns of cathepsin B from lung carcinoma and matched lung were similar, particularly with regard to 2 major intermediate acidic enzyme pI forms designated as I and II (pIapp of 5.10 and 4.93 in tumors, and 5.11 and 4.94 in lungs, respectively). The slightly acidic cathepsin B pI forms (pIapp 5.47-5.19) in squamous-cell lung carcinoma (SQCLC) were significantly more numerous than such enzyme pI forms in lungs. The numbers of the highly acidic cathepsin B pI forms (pIapp 4.82-4.33) were significantly higher in SQCLC and lung adenocarcinoma (ACL) than in matched lung. The activity distribution percentage in the set of highly acidic cathepsin B pI forms was significantly higher in SQCLC and ACL than in matched lung. We also observed that cathepsin B from SQCLC and matched lung was fully recoverable by IEF from inhibition by leupeptin. Using the cysteine-proteinase-specific inactivator E-64, we revealed by IEF that some cathepsin B isoforms (charge forms) from SQCLC were more resistant to inactivation by this compound than the corresponding enzyme isoforms from lungs. After IEF, the enzyme isoforms apparently lost their resistance to E-64. Our results indicate that the pool of multiple charge forms of cathepsin B in SQCLC and ACL is different from that in the lung, and also that there may be an increased level of loose complexes between cathepsin B and some proteins or polypeptides in SQCLC compared to the lung.

Adenocarcinoma↗

Extracellular volume fraction and diffusion characteristics during progressive ischemia and terminal anoxia in the spinal cord of the rat.

Extracellular space (ECS) volume fraction (alpha), ECS tortuosity (lambda), and nonspecific uptake (k'), three parameters affecting the diffusion of substances in nervous tissue, were studied during ischemia and anoxia in the rat spinal cord gray matter in vivo. Progressive ischemia evoked by exsanguination, as well as anoxia evoked by respiratory or cardiac arrest, produced prominent extracellular K+ and pH changes closely related to a decrease in blood pressure and amplitude of field potentials. With use of ion-selective microelectrodes, the changes in the diffusion parameters were measured by quantitative analysis of concentration-time profiles of tetramethylammonium (TMA+) applied by iontophoresis concomitantly with ionic shifts. Under normoxic conditions (in rats with blood pressure of 80-110 mm Hg) diffusion parameters in the dorsal horn gray matter at depth 500-900 microns were as follows: alpha = 0.20 +/- 0.019, lambda = 1.62 +/- 0.12, k' = 4.6 +/- 2.5 x 10(-3) s-1 (mean +/- SD, n = 39). Extracellular K+, pH, and diffusion properties gradually changed during progressive ischemia. As the blood pressure fell to 50-60 mm Hg and field potential amplitude to 20-60%, K+ rose to 6-12 mM, pHe fell by approximately 0.05-0.1 pH unit, and volume fraction of the ECS significantly decreased, to alpha = 0.16 +/- 0.019 (n = 22). Even though the tortuosity remained virtually constant, the nonspecific uptake significantly decreased to k' = 3.4 +/- 1.8 x 10(-3) s-1. As the blood pressure fell to 20-30 mm Hg and field potential amplitude to 0-6%, K+ rose to 60-70 mM, pHe fell by approximately 0.6-0.8 pH unit, and all three diffusion parameters significantly changed. The ECS volume fraction decreased to alpha = 0.05 +/- 0.021, tortuosity increased to lambda = 2.00 +/- 0.24, and TMA+ uptake decreased to k' = 1.5 +/- 1.6 x 10(-3) s-1 (n = 12). No further increase in extracellular K+ or changes in the alpha were found during and up to 120 min after the death of the animal. However, there was a further significant increase in lambda = 2.20 +/- 0.14 and decrease in k' = 0.4 +/- 0.3 x 10(-3) s-1 (n = 24). The acid shift reached its maximum level at approximately 5-10 min after respiratory arrest and then the pHe gradually increased by approximately 0.2 unit. Full recovery to "normoxic" diffusion parameters was achieved after reinjection of the blood or after an injection of noradrenaline during severe ischemia, if this resulted in a rise in blood pressure above 80 mm Hg and a decrease in extracellular K+ below 12 mM.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

[The Bayesian statistical theory in the diagnosis of malignant and non-malignant diseases of the lung, pleura and mediastinum].

The Bayesian algorithm was used to assess the probable diagnosis in 1262 patients with a recently diagnosed finding on X-rays of the chest and the results were compared with the final diagnosis. The patients were with regard to the X-ray picture divided into 9 groups: hilar, solitary, multiple, segmental, non-segmental, cavity, diffuse, pleural and mediastinal lesions. Using the Bayesian algorithm, commonly accessible factors were processed: age, sex, case-history, cigarette smoking, red cell sedimentation rate, number of leucocytes and diameter of solitary parenchymatous lesions and the impact of these factors for assessment of probability of a malignant or non-malignant lesion was evaluated. The reliability in different X-ray lesions was within the range of 84.2% to 92.4%. The authors evaluated also tests of sensitivity, specificity, the reliability of forecast of a positive and negative result, which confirmed the differences in the different groups which showed evaluated. Analysis of the results, provided evidence that the Bayesian algorithm is a promising objective method for the forecast of a malignant or non-malignant diagnosis in patients with a newly diagnosed X-ray finding of the chest.

Adolescent↗

[Smoking and the development of lung cancer].

It is generally accepted that cigarette smoking is the most important lung cancer risk factor to which 80-90% of lung cancer mortality in advanced countries with a high tobacco consumption is ascribed. In the Czech Republic the increase of tobacco consumption since the end of the Second World War was arrested at the end of the sixties. Since the beginning of the seventies the cigarette consumption did not change substantially and varied close to 2500 cigarettes per inhabitant above 15 years. No reliable data are so far available in the Czech Republic on the development of the prevalence of smoking in different age groups of the population. According to several non-representative surveys in different districts it may be assumed that in men aged 25-64 years in the course of the last 20 years the ratio of male smokers declined probably, while it increased in female smokers. Due to the long "latency" between exposure to noxious substances and the manifestation of lung cancer which amounts to two, three or more decades, we have to evaluate the present mortality rate from lung cancer as a reflection of carcinogenic factors (above all cigarette smoking) in the more remote past. When evaluating the trend of mortality from lung cancer in the Czech Republic during 1953-1989, throughout that period the values were much higher in men that in women and the trend in the two sexes was quite different. In men the mortality rate from lung cancer increased in 1953-1967 to three times the initial value, while in women the levels remained low.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Model of distribution of anticancer agents transplanted into the brain tissue using the random walk method.

A Monte-Carlo approach to analysis of dispersion in the tissue of a locally administered drug is presented. The distribution of a drug in the tissue is simulated as a distribution of randomly walking particles. The approach is demonstrated on a simple situation for which both experimental results and an analytical solution are known. The approach can be used in situations, where common numeric methods are difficult to use, especially for analyses of drug transport in an inhomogeneous space, and problems with complex boundary conditions, e.g. in analyses of dispersion of anticancer agents locally applied into tumours.

Animals↗

[Environmental monitoring and biological monitoring of young people exposed to nonoccupational levels of formaldehyde, toluene and other hydrocarbons].

During the period of 1983-1985, in two of apprentice schools of P. town the health disorders were investigated in the total of 82 apprentices 15-18 years old from the environment with elevated concentrations of formaldehyde and toluene. The study was contrasted with a control total of 42 apprentices. Cytogenetical examination has been performed, and selected immunological parameters in both blood serum and saliva have been assessed with red and white blood cells counts including differential formula of white blood cells. In addition, the atmospheric toxicity of formaldehyde and vapours of organic solvents (toluene, xylene, varnish naphtha) was measured. A single biological exposure test has been performed for the detection toluene. Statistically significant were differences in occurrence of cell chromosomal aberrations between the group of long term formaldehyde and toluene exposure (averagely 3.53% ABB) and controls (2.21% ABB) as obtained in 1983 and 1984, and so were differences between the long term-to-toluene exposed group (3.30% ABB) and the above mentioned control group as obtained in 1984. No similar results were stated between the long term-to-formaldehyde exposed (3.07% ABB) and control (2.55% ABB) groups in 1985. The main evidence consisted in finding the genotoxical/clastogenic effect of observed agents associated with mainly chromosomal abnormalities of chromatide type. It outflowed from the determination of selected serum proteins (Ig and acute phase proteins) and salivary lysozyme that the group under the combined influence of formaldehyde and toluene showed significantly lower IgG and higher alpha-1-antitrypsin (A1AT). The group at risk of toluene was characteristical in elevated concentrations of alpha-2-macroglobulin (A2M) and A1AT. Most pronounced changes in first year had been revealed through the evaluation of the influence of the duration at risk (significant decrease in IgA and prealbumin, and the increase in A2M and A1AT). The infectious disease as experienced 2 month prior the collection resulted in a significant decrease of IgM, A2M and A1AT in risky groups in individuals with infection in anamnesis. Salivary lysozyme concentration of apprentice environmentally exposed to formaldehyde in the noon showed the decrease, whereas its increase occurred in controls with the difference on 5% significancy level. Blood count assessements showed no significant differences between the investigated values as well as any were assessed between the incidence of health disorders of apprentices and their correspondance to the given group.

Adolescent↗