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1.
Am J Hematol ; 99(4): 745-750, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38264829

ABSTRACT

Profound immune dysregulation and impaired response to the SARS-CoV-2 vaccine put patients with chronic lymphocytic leukemia (CLL) at risk of severe COVID-19. We compared humoral memory and T-cell responses after booster dose vaccination or breakthrough infection. (Green) Quantitative determination of anti-Spike specific antibodies. Booster doses increased seroconversion rate and antibody titers in all patient categories, ultimately generating humoral responses similar to those observed in the postinfection cohort. In detail, humoral response with overscale median antibody titers arose in >80% of patients in watch and wait, off-therapy in remission, or under treatment with venetoclax single-agent. Anti-CD20 antibodies and active treatment with BTK inhibitors (BTKi) represent limiting factors of humoral response, still memory mounted in ~40% of cases following booster doses or infection. (Blue) Evaluation of SARS-CoV-2-specific T-cell responses. Number of T-cell functional activation markers documented in each patient. The vast majority of patients, including those seronegative, developed T-cell responses, qualitatively similar between treatment groups or between vaccination alone and infection cases. These data highlight the efficacy of booster doses in eliciting T-cell immunity independently of treatment status and support the use of additional vaccination boosters to stimulate humoral immunity in patients on active CLL-directed treatments.


Subject(s)
COVID-19 , Leukemia, Lymphocytic, Chronic, B-Cell , Humans , SARS-CoV-2 , Leukemia, Lymphocytic, Chronic, B-Cell/drug therapy , COVID-19 Vaccines , Antibodies , Interleukin-2 Receptor alpha Subunit , Immunity, Cellular , Antibodies, Viral , Vaccination
2.
J Immunol Res ; 2017: 7304658, 2017.
Article in English | MEDLINE | ID: mdl-29392143

ABSTRACT

Humoral deficiencies represent a broad group of disorders. The aim of the study was to compare the levels of antibodies against pneumococcal capsular polysaccharides (anti-PCP) and natural anti-galactosyl (anti-Gal) antibodies in (1) patients with chronic lymphocytic leukaemia (CLL), (2) patients with common variable immunodeficiency (CVID), and (3) a healthy population and to explore their diagnostic and prognostic potential. Serum immunoglobulin levels and levels of anti-Gal IgG, IgA, and IgM and anti-PCP IgG and IgG2 were determined in 59 CLL patients, 30 CVID patients, and 67 healthy controls. Levels of IgG, IgA, IgM, anti-Gal IgA, anti-Gal IgM, and anti-PCP IgA were lower in CLL and CVID patients than in healthy controls (p value for all parameters < 0.0001). Decrease in the levels of IgA, IgM, anti-Gal IgA, and anti-PCP IgA was less pronounced in the CLL group than in the CVID group. IgA decline, anti-Gal IgA, anti-PCP IgA, and anti-PCP IgG2 were negatively correlated with CLL stage. We devise the evaluation of anti-Gal antibodies to be a routine test in humoral immunodeficiency diagnostics, even in cases of immunoglobulin substitution therapy. Significant reductions, mainly in anti-Gal IgA, IgM, and anti-PCP IgA levels, may have prognostic importance in CLL patients.


Subject(s)
Bacterial Capsules/immunology , Common Variable Immunodeficiency/immunology , Pneumococcal Infections/immunology , Streptococcus pneumoniae/immunology , Adolescent , Adult , Aged , Aged, 80 and over , Antibodies, Bacterial/blood , Autoantibodies/blood , Biomarkers/blood , Common Variable Immunodeficiency/diagnosis , Female , Galactosylceramides/immunology , Humans , Immunity, Humoral , Immunoglobulin A/blood , Immunoglobulin G/blood , Immunoglobulin M/blood , Male , Middle Aged , Pneumococcal Infections/diagnosis , Prognosis , Young Adult
3.
Thromb Res ; 136(6): 1082-6, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26467608

ABSTRACT

INTRODUCTION: Venous thromboembolism (VTE) is a major cause of morbidity and mortality in patients (pts) with malignant tumors. Increased risk of VTE is well described in a variety of hematologic malignancies; however, data regarding VTE in chronic lymphocytic leukemia (CLL) is very limited. PATIENTS AND METHODS: We retrospectively analyzed clinical and laboratory data of 346 consecutive pts with CLL followed up at 4th Department of Internal Medicine - Hematology, University Hospital, Hradec Kralove, Czech Republic, diagnosed between 1999 and 2011 (males, 64%; median age, 64 years; low/intermediate/high Rai modified risk in 41/47/12%). RESULTS: After a median follow-up of 72 months (range, 26-138), at least one episode of VTE occurred in 38 patients (11%). VTE developed after a median of 34 months from CLL diagnosis. Incidence of VTE was 1.67% per patient year of follow-up. There was a high proportion of unfavourable prognostic factors (advanced Rai stages, unmutated IgVH genes, unfavourable cytogenetics) in pts with VTE. The presence of 0/1/2/3 additional risk factors for VTE was identified in 2/16/14/6 patients. The most common risk factors for VTE besides age (n=24) were corticosteroid therapy (n=13), other malignancies (n=9) and obesity (n=7). Recurrence of VTE was diagnosed in 7 pts. Performance status ≥ 2 and inherited thrombophilia were significant risk factors for VTE development in univariate and multivariate analysis. VTE was not associated with shorter overall survival. CONCLUSION: Based on our results, VTE is a relatively frequent complication in patients with CLL. Although most patients had other known risk factors for VTE including CLL treatment, 29% had no risk factors or only age ≥ 60 years. These findings demonstrate the possible role of CLL in the development of VTE.


Subject(s)
Leukemia, Lymphocytic, Chronic, B-Cell/complications , Venous Thromboembolism/etiology , Adult , Aged , Aged, 80 and over , Female , Humans , Leukemia, Lymphocytic, Chronic, B-Cell/mortality , Male , Middle Aged , Risk Factors
4.
Neoplasma ; 61(5): 585-92, 2014.
Article in English | MEDLINE | ID: mdl-25030442

ABSTRACT

Elevated levels of circulating angiogenic cytokines and increased expression of genes encoding angiogenic factors have been reported in recent years in patients with chronic lymphocytic leukemia (CLL) but data regarding prognostic and predictive significance are still limited. Therefore, in the present study based upon our prior pilot results, we measured mRNA expressions of angiopoietin-2 (Ang-2), fibroblast growth factor-2 (FGF-2) and endoglin (CD105) by reverse transcription quantitative PCR in purified CD19+ cells from 70 untreated CLL patients (median age, 63 years; males, 64%; Rai III/IV stages, 29 %; unmutated IgVH genes, 60 %) and evaluated their possible association with established prognostic factors and clinical course of the disease. Higher expression of Ang-2 was significantly associated with unmutated IgVH genes (n = 55, p = 0.003). Higher CD105 expression was significantly associated with unmutated IgVH genes (n = 55, p < 0.001), high CD38 expression (n = 66, p = 0.022), high ZAP-70 expression (n = 66, p = 0.010), Rai stage I-IV (n = 70, p < 0.001), progressive clinical course of CLL (n = 70, p = 0.001) and shorter time to treatment (n = 70; p < 0.001). Expression of FGF-2 was not significantly associated with any of the prognostic markers. These results indicate that elevated expression of Ang-2 and in particular CD105 by CLL cells is associated with unfavorable prognostic features and clinical outcome; thus, both cytokines appear to play an important role in biology and progression of CLL and warrant further investigation.


Subject(s)
Angiopoietin-2/genetics , Antigens, CD/genetics , Fibroblast Growth Factor 2/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/metabolism , RNA, Messenger/analysis , Receptors, Cell Surface/genetics , Adult , Aged , Aged, 80 and over , Endoglin , Female , Humans , Immunoglobulin Heavy Chains/genetics , Immunoglobulin Variable Region/genetics , Male , Middle Aged , Mutation
5.
Vnitr Lek ; 59(9): 782-93, 2013 Sep.
Article in Czech | MEDLINE | ID: mdl-24073950

ABSTRACT

INTRODUCTION: Search for new prognostic markers in order to improve prognostic accuracy and predict clinical outcome at the time of dia-gnosis has recently become one of the major trends in chronic lymphocytic leukemia (CLL). PATIENTS AND METHODS, AIM OF STUDY: The aim of our study was assessment of selected markers of apoptosis and angiogenesis and their potential as new prognostic factors. We evaluated serum levels of tumor necrosis factor α (TNFα) and transforming growth factor ß 1 (TGFß1) using commercially available enzyme linked immunosorbent assay; furthermore, we quantified expression of type II receptor for transforming growth factor beta (TGFßRII) and type 2 receptor for fibroblast growth factor 2 (FGFR2) on CLL cells using flow cytometry analysis in 75 previously untreated patients with CLL (47 males and 28 females, median age, 65 years, range 38- 82) and healthy donors. RESULTS: We found significantly elevated TNFα in patients with CLL compared to the control group (p < 0.0001); high expression of TNFα was associated with unfavourable prognosis: significantly higher concentrations were found in patients with Rai highrisk group compared to low and intermediate-risk group (p = 0.0008 and p = 0.0097), with high serum ß2- microglobulin (p = 0.045), massive lymphadenopathy (p = 0.0083), unmutated genes for variable region of immunoglobulin heavy chain (IgVH) (p = 0.041) and unfavourable cytogenetic aberrations (p = 0.0014). In addition, patients with progressive CLL had significantly higher TNFα than those with stable clinical course (p = 0.0009); time to treatment was significantly shorter in patients with higher TNFα (p = 0.0049). Higher TGFß1 concentrations were associated with favourable subgroups: with Rai low  risk group compared to high risk group (p = 0.011), patients without massive lymphadenopathy (p = 0.041), patients with mutated IgVH (p = 0.012) and ZAP 70 negativity (zeta associated protein of 70 kilodaltons) (p = 0.044). Patients with progressive CLL had significantly lower TGFß1 levels than those with stable course (p = 0.0014) and time to treatment was significantly longer in patients with higher TGFß1 (p = 0.016). Patients with Rai high risk group had significantly lower TGFßRII expression than those with low  risk group (p = 0.022). The prognostic significance of FGFR2 was not found. Significant and independent prognostic factors for overall survival were high serum concentrations of TNFα and massive lymphadenopathy (p = 0.036, resp. p = 0.047). CONCLUSION: Based on our results, TNFα and TGFß1 possess prognostic significance in CLL; further research in this direction may also be important therapeutically, because these signal pathways could serve as possible treatment targets.


Subject(s)
Apoptosis/physiology , Biomarkers, Tumor/blood , Leukemia, Lymphocytic, Chronic, B-Cell/blood , Neovascularization, Pathologic/blood , Adult , Aged , Aged, 80 and over , Disease Progression , Female , Flow Cytometry , Humans , Immunoglobulin Heavy Chains , Leukemia, Lymphocytic, Chronic, B-Cell/drug therapy , Male , Middle Aged , Prognosis , Protein Serine-Threonine Kinases/blood , Receptor, Fibroblast Growth Factor, Type 2/blood , Receptor, Transforming Growth Factor-beta Type II , Receptors, Transforming Growth Factor beta/blood , Reference Values , Transforming Growth Factor beta1/blood , Tumor Necrosis Factor-alpha/blood , ZAP-70 Protein-Tyrosine Kinase
6.
Int J Biol Macromol ; 48(2): 319-25, 2011 Mar 01.
Article in English | MEDLINE | ID: mdl-21145345

ABSTRACT

The interaction of a copper complexes containing Schiff bases with calf thymus (CT) DNA was investigated by spectroscopic methods. UV-vis, fluorescence and CD spectroscopies were conducted to assess their binding ability with CT DNA. The binding constants K have been estimated from 0.8 to 9.1×10(4) M(-1). The percentage of hypochromism is found to be over 70% (from spectral titrations). The results showed that the copper(II) complexes could bind to DNA with an intercalative mode. Synergic action of Cu(II) complexes with ascorbic acid against Candida albicans induced the generation of free radicals and increased (more than 60 times) antimicrobial effect of these complexes.


Subject(s)
Anti-Infective Agents/pharmacology , Copper/metabolism , Copper/physiology , DNA/metabolism , Schiff Bases/metabolism , Schiff Bases/pharmacology , Animals , Ascorbic Acid/pharmacology , Candida albicans/drug effects , Candida albicans/growth & development , Cattle , Circular Dichroism , Electrons , Intracellular Space/metabolism , Microbial Sensitivity Tests , Microscopy, Fluorescence , Plasmids , Reactive Oxygen Species , Spectrometry, Fluorescence , Titrimetry
7.
Vnitr Lek ; 54(5): 452-6, 2008 May.
Article in Czech | MEDLINE | ID: mdl-18630625

ABSTRACT

Diabetes mellitus is a frequent cause of renal insufficiency. Renal insufficiency is associated with both haemorrhagic manifestations primarily caused by platelet functional disorders, and states of hypercoagulation resulting from significant hyperfibrinogenemia. Fibrinolysis is either increased or, often, decreased. Changes in haemostasis in renal insufficiency have been dealt with by many authors in relevant literature. However, the final stage of renal insufficiency is rather dominated by haemorrhagic diathesis. It is manifested by skin haemorrhage, mucosal manifestations, but also by retroperitoneal and cerebral haemorrhage. The main cause of a haemorrhagic condition is platelet dysfunction combined with anticoagulation and antiplatelet therapy which is used in dialysis. Platelet function disorders are provoked by acquired thrombocytopaenia and result in a disorder in the interaction between the blood vessel wall and the platelet. Dialysis suppresses platelet abnormalities only temporarily by suppressing uremic toxins provoking platelet disorders. On the other hand, dialysis may cause prothrombotic activity. Changes in haemostasis in type 2 diabetes mellitus form part of the insulin resistance syndrome and induce prothrombotic condition due to decreased fibrinolysis.


Subject(s)
Blood Coagulation , Diabetes Mellitus, Type 2/blood , Diabetic Nephropathies/blood , Renal Insufficiency/blood , Blood Platelets/physiology , Diabetes Mellitus, Type 2/complications , Diabetes Mellitus, Type 2/therapy , Diabetic Nephropathies/complications , Hemorrhagic Disorders/blood , Hemorrhagic Disorders/etiology , Hemostasis , Humans , Renal Insufficiency/complications , Thrombosis/blood , Thrombosis/etiology
8.
J Basic Microbiol ; 44(2): 122-8, 2004.
Article in English | MEDLINE | ID: mdl-15069671

ABSTRACT

The influx of chlorides into Trichoderma viride vegetative submerged mycelium was measured by means of the radionuclide (36)Cl(-). It was found that the (36)Cl(-) influx was time-dependent (the steady-state was established with t(1/2 )= 25 min at 25 degrees C), pH-dependent (with pH optimum between 4-5.5), temperature-dependent (at about 15 degrees C), and concentration-dependent (K(M)(Cl(-))) = 47.6 +/- 4.2 micromol x l(-1); J(max) = 11.5 +/- 0.7 pmol(Cl(-)) x min(-1). mg(dry mass) (-1)). The (36)Cl(-) influx was inhibited by Br(-) but not F(-), I(-), SO(4)(2-), HPO(3)(2-) and HCO(3)(-). The presence of vanadate (P-type ATPase inhibitor) moderately stimulated the (36)Cl(-) influx but the presence valinomycin (electrogenic K(+) ionophore), salicylate (known to release Ca(2+) from Trichoderma viride internal stores) were without effect on the (36)Cl(-) influx. The results suggest that the (36)Cl(-) influx is mediated by a carrier and that the transport is electroneutral, probably Cl(-)/OH(-) antiport.


Subject(s)
Chlorides/metabolism , Trichoderma/metabolism , Biological Transport/drug effects , Bromine/pharmacology , Mycelium/metabolism , Time Factors , Vanadates/pharmacology
9.
Mol Cell Biochem ; 231(1-2): 15-22, 2002 Feb.
Article in English | MEDLINE | ID: mdl-11952157

ABSTRACT

A boron-containing antibiotic, boromycin (BM), was found to influence the Ca2+ homeostasis in both excitable and non-excitable cells. In non-excitable cells (human erythrocytes and leucocytes) it inhibited the resting passive 45Ca2+ transport in 10(-6)-10(-5) mol/L concentrations. In human erythrocytes, the passive 15Ca2+ transport induced by the presence of 1 mmol/L NaVO3 was inhibited by boromycin (90% inhibition) as well. The inhibitory effect of BM on the NaVO3-induced passive 45Ca2+ transport was diminished in the presence of inhibitory concentrations of nifedipine (10 micromol/L -60% inhibition) or of those of K+o (75 mmol/L -20% inhibition). On the other hand, in rat brain synaptosomes, and rat cardiomyocytes, BM stimulated the passive 45Ca2+ transport in 'resting' cells at similar concentrations. In rat cardiomyocytes the stimulation was transient. The stimulatory effect on the passive 45Ca2+ transport in rat brain synaptosomes was accompanied with the increase of cytoplasmic Ca2+ concentration measured by means of the entrapped fluorescent Ca2+ chelator fura-2. The stimulatory effect of BM was diminished when synaptosomes were pre-treated with veratridine (10 micromol/L) which itself stimulated the passive 45Ca2+ transport. At saturating concentrations of veratridine, no stimulatory effect of BM was observed. These results could be explained by the indirect interaction of BM with both Ca2+ and Na+ transport systems via transmembrane ionic gradients of monovalent cations and could be useful in determining whether the cells belong to excitable, or non-excitable cells.


Subject(s)
Borates/pharmacology , Calcium/metabolism , Homeostasis/drug effects , Animals , Dose-Response Relationship, Drug , Erythrocytes/drug effects , Erythrocytes/metabolism , Female , Heart/drug effects , Humans , Leukocytes/drug effects , Leukocytes/metabolism , Male , Rats , Synaptosomes/drug effects
10.
Gen Physiol Biophys ; 20(2): 131-44, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11519685

ABSTRACT

Azalomycin F (AMF), a macrocyclic lactone antibiotic, in concentrations of 10(-5) g/ml (10(-6) - 10(-5) mol/l) was found to stimulate both the 45Ca2+ influx and efflux in intact Trichoderma viride submerged mycelium and in cells of Saccharomyces cerevisiae without having Ca2+ ionophoric properties. AMF also inhibited ATP-dependent Ca2+ uptake in membrane fractions prepared from T. viride submerged mycelium. 45Ca2+ which had been accumulated in membrane fractions in an ATP-dependent manner was released upon addition of AMF. This release was observed in light organellar fractions (LOF) of S. cerevisiae and of T. viride submerged mycelium and, to a small extent, in heavy organellar fraction (HOF) of S. cerevisiae. No Ca2+ releasing effect of AMF was observed in HOF from T. viride submerged mycelium. In S. cerevisiae expressing Ca2+-dependent photoprotein aequorin, AMF induced transients of luminescence which reflect changes in the cytoplasmic Ca2+ concentration. The results suggest that the stimulation by AMF of the Ca2+ efflux from the mycelium (cells) could be explained by an increase of the cytoplasmic Ca2+ concentration due to the release of Ca2+ from microsomal membranes or to the stimulation of Ca2+ influx.


Subject(s)
Anti-Bacterial Agents/pharmacology , Calcium/metabolism , Saccharomyces cerevisiae/drug effects , Trichoderma/drug effects , Biological Transport/drug effects , Cell Fractionation , Egtazic Acid/pharmacology , Homeostasis/drug effects , Homeostasis/physiology , Kinetics , Macrolides , Organelles/drug effects , Organelles/physiology , Saccharomyces cerevisiae/physiology , Subcellular Fractions/metabolism , Trichoderma/physiology
11.
Arch Microbiol ; 175(1): 32-40, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11271418

ABSTRACT

Glutamic acid decarboxylase (GAD) activity was measured in homogenates of conidia and both submerged and aerial mycelia of Trichoderma viride. The GAD activity in conidia had a temperature optimum at 30 degrees C and a pH optimum at pH 4. GAD was stimulated by EDTA (2 mM) and was insensitive to treatment with calmodulin antagonists calmidazolium (10 microM) or phenothiazine neuroleptics (60 microM). Cyclosporin A (up to 300 microM) partially inhibited GAD in the homogenate, but not in the supernatant obtained after centrifuging the homogenate. Attempts to release GAD activity from the homogenate using high ionic strength, detergents, or urea failed. Freezing-thawing led to the partial increase of activity in the conidial homogenate. These results indicate that GAD is a membrane-bound enzyme. The highest specific activity of GAD was present in the mitochondrial/vacuolar organellar fraction. Germination of conidia in the submerged culture led to a temporary decrease in GAD activity. After prolonged cultivation, the activity displayed quasi-oscillatory changes. The stationary state was characterized by a high GAD activity. The presence of gamma-aminobutyric acid in the submerged mycelia was demonstrated. In surface culture in the dark, GAD activity increased in a monophasic manner until conidia formation. The illumination of dark-cultivated mycelia by a white-light pulse caused a dramatic increase in GAD activity. Light-induced changes were not observed in mutants with delayed onset of conidiation. In the dark or upon illumination by light pulse, the increase of GAD activity preceded the appearance of conidia. Thus, GAD activity in T. viride is closely associated with its developmental status and may represent a link between differentiation events and energy metabolism.


Subject(s)
Glutamate Decarboxylase/metabolism , Trichoderma/enzymology , Cyclosporine/pharmacology , Edetic Acid/pharmacology , Enzyme Inhibitors/pharmacology , Freezing , Glutamate Decarboxylase/antagonists & inhibitors , Glutamate Decarboxylase/isolation & purification , Hydrogen-Ion Concentration , Imidazoles/pharmacology , Light , Organelles/enzymology , Phenothiazines/pharmacology , Spores, Fungal , Temperature , Trichoderma/drug effects , Trichoderma/growth & development , gamma-Aminobutyric Acid/metabolism
12.
Can J Microbiol ; 46(4): 312-24, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10779867

ABSTRACT

The properties of both Ca2+ influx and efflux in the mycelium during the life cycle of Trichoderma viride were studied by means of 45Ca2+ and by X-ray fluorescence spectroscopy measurements. The properties of the 45Ca2+ influx and effluxes indicate that they are mediated by different transport systems. The Ca2+ influx could be mediated by an electrogenic Ca2+/nH+ antiport, or by an Ca2+ uniport system. Both Ca2+ influx and efflux were stimulated by the uncouplers (and the treatment leading to the suppression of energy metabolism) and by azalomycin F, an antifungal agent. Salicylate stimulated the Ca2+ efflux, but inhibited the Ca2+ influx. In the isolated preparation of crude vacuolar/mitochondrial fraction, salicylate induced the Ca2+ release, as did A23187. Azalomycin F moderately released Ca2+ from the microsomal fraction. On the other hand, uncouplers did not release Ca2+ from the isolated organelles, but inhibited to a different extent the ATP-dependent and -independent Ca2+ influx. The results could be explained in terms of the capacitative Ca2+ influx mechanism. The rate of 45Ca2+ influx, or of the 40Ca2+ content, was maximal after about 30 h of submerged cultivation, and then decreased. The results show that loading of internal Ca2+ stores occurs in the early stages of the development of mycelium only, and the Ca2+ influx mechanism is developmentally down-regulated, being almost nonexistent during its later stages. In older mycelium, growth seems to be autonomous of the extracellular Ca2+ until the onset of conidiation.


Subject(s)
Calcium/metabolism , Trichoderma/growth & development , Anti-Bacterial Agents/pharmacology , Calcimycin/pharmacology , Calcium/antagonists & inhibitors , Calcium Radioisotopes , Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone/pharmacology , Hydrogen-Ion Concentration , Ionophores/pharmacology , Macrolides , Salicylates/pharmacology , Spectrometry, X-Ray Emission , Subcellular Fractions/metabolism , Temperature , Time Factors , Trichoderma/metabolism , Uncoupling Agents/pharmacology
13.
FEMS Microbiol Lett ; 171(2): 161-5, 1999 Feb 15.
Article in English | MEDLINE | ID: mdl-10077840

ABSTRACT

The transport of radioactively labelled uracil into submerged mycelium of T. viride was measured by means of a membrane filtration technique. It was found to be time-dependent (up to 90 min) and concentration-dependent (up to 8 mmol l-1). Its concentration dependence was biphasic and consisted from the saturatable part (at the uracil concentration below 0.2 mmol l-1) with KM = 0.08 +/- 0.02 mmol l-1 and Vmax = 1.74 +/- 0.3 nmol (mg dry wt.)-1 h-1, and from the region at higher uracil concentration which showed only a weak saturatability with the substrate. The transport measured in the saturatable part of the curve was also pH- and temperature-dependent. The optimal pH was between 5.4 and 6.4 and the optimal temperature was at 37 degrees C. The activation energy of 54 kJ mol-1 and the temperature quotient of Q10 = 2.1 could be calculated from the temperature dependence. The entry of uracil was in part inhibited by nucleobases and their analogues, nucleosides, nucleotides and amino acids. The inhibitors had similar inhibitory efficiency about 50% at 0.2 mmol l-1. 3,3',4',5-tetrachlorosalicylanilide (TCS), the uncoupling agent, significantly inhibited the uracil transport, but its inhibitory efficiency decreased upon increasing the uracil concentration. Ionophore antibiotics valinomycin and monensin also inhibited the uracil transport. Inhibitors of RNA-polymerase, rifamycin and rifampicin were without effect. The results suggest that at low uracil concentrations (below 0.2 mmol l-1), its transport is mediated by a carrier and is driven by the electrochemical potential of protons. At higher uracil concentrations, the transport may be driven by the concentration difference of uracil with the contribution of the protonmotive force. It is feasible that inhibitors of uracil transport tested exert their inhibition by the dissipation of the driving force rather than by the direct competition with the substrate-binding site.


Subject(s)
Trichoderma/metabolism , Uracil/pharmacokinetics , Biological Transport/drug effects , Biological Transport/physiology , Carrier Proteins/metabolism , Cytosine/analogs & derivatives , Cytosine/pharmacology , Fungal Proteins/metabolism , Histidine/pharmacokinetics , Ionophores/pharmacology , Lysine/pharmacokinetics , Methanol/pharmacology , Monensin/pharmacology , Solvents/pharmacology , Thymidine/pharmacokinetics , Tritium , Tyrosine/pharmacokinetics , Valinomycin/pharmacology
14.
Folia Microbiol (Praha) ; 42(2): 125-9, 1997 Apr.
Article in English | MEDLINE | ID: mdl-18454332

ABSTRACT

The rate of the(45)Ca(2+) uptake by the submergedTrichoderma viride mycelium increased with the age of the culture from 6 h until a maximum which was reached at about 30 h, and then decreased until the uptake was virtually zero. The decrease in the rate of the(45)Ca(2+) uptake was accompanied by an increase of mycelial mass. The uptake rate could not be reactivated upon substituting the medium for a fresh one, without or with dilution of the mycelium. The results suggest that the rate of(45)Ca(2+) uptake reflects the biological age of the submerged culture. The surface-cultivated mycelium took up(45)Ca(2+) proportionally with time. The autoradiography of colonies showed that(45)Ca(2+) was distributed homogeneously throughout the mycelium during vegetative growth while conidiation was accompanied by a massive accumulation of(45)Ca(2+) in conidia.

15.
Cesk Psychiatr ; 89(5): 300-5, 1993 Oct.
Article in Slovak | MEDLINE | ID: mdl-8269525

ABSTRACT

Process-oriented psychotherapy /POP/ is an eclectic trend. It is based on Jungian psychology but is inspired also by some dynamic schools and oriental philosophy, in particular Taoism. The essence of the psychotherapeutic method is work with signals in the communication channels. By their reinforcement the personality attains contact with the secondary process behind the borderline of identity. In addition to psychopathological indications it uses somatic symptoms and diseases which are considered a purposeful message of the dream body.


Subject(s)
Psychotherapy/methods , Humans
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