Evaluation of the Hepatoprotective Effect of L-Ascorbate 1-(2-Hydroxyethyl)-4,6-Dimethyl-1,2-Dihydropyrimidine-2-One Upon Exposure to Carbon Tetrachloride.

Hepatoprotective properties of a new pyrimidine derivative - L-ascorbate 1-(2-hydroxyethyl)-4,6-dimethyl-1,2-dihydropyrimidine-2-one, synthesized on the basis Xymedon, were assessed in white rats exposed to CCl4. The compound under study administered prior to exposure to CCl4 reduced the deviation of biochemical parameters from reference values and severity of structural and morphological changes in liver, when compared to the control. Hepatoprotective properties of the studied compound were more pronounced than those of Xymedon.

[EPR study of iron status in human body during intensive physical activity].

The iron metabolism was studied in serum blood samples collected from 26 professional sportsmen undergoing intensive physical exercises using EPR combined with haematological and biochemical laboratory tests. Only 23% of EPR spectra (n = 6) were practically normal while in the rest spectra additional abnormal absorption lines were detected. Presumably, the significant portion of new signals may be caused by different cytochromes. Moreover, the anisotropic signals with g1 approximately equal to 2.02; g2 approximately equal to 1.94 and g3 approximately equal to 1.86 registered in some spectra pointed to the sulfur-iron centers. There was nearly linear correlation between the concentration of Fe3+ in transfferin (Fe(3+)-Tf) obtained from the EPR spectra and the serum iron concentration measured by absorption photometry both for sportsmen and controls (healthy individuals and patients with different diseases). At equal serum iron concentrations the Fe(3+)-Tf level was higher in sportsmen than that in controls. The Pearson correlation coefficient (r) for Fe(3+)-Tf and serum iron values was equal to 0.89 in sportsmen versus r = 0.97 in controls. Additional new lines in serum EPR spectra of professional sportsmen prove the suitability of EPR assay for scheduled medical exams since routinebiochemical and haematological tests are insufficient to discover all abnormalities in iron metabolism under intensive physical exercises.

[Electron paramagnetic resonance study of blood of anemic patients with urological cancer].

Changes in Fe(3+)-transferrin (Fe(3+)-Tf) and Cu(2+)-ceruloplasmin (Cu(2+)-Cp) concentrations in venous blood sampled from anemic patients with urinary bladder and kidney cancer in I-IV stages were investigated using electron paramagnetic resonance spectroscopy. It was established that at malignancy-associated anemia the paramagnetic Fe3+ ion concentration in transferrin is below a norm, while in anemic non-oncology patients the Tf iron saturation is normal. Moreover, in patients with malignancy-associated anemia the Cu(2+)-Cp average value is nearly twice as large as that for healthy volunteers (confidence probability P). It was shown that simultaneous EPR measuring of paramagnetic centers (such as Fe(3+)-Tf and Cu(2+)-Cp) in blood of anemic patients can be used as a biomarker for urological cancer diagnosis even at early stages of the growth of a malignant tumor.

Correlation of T cell response and bacterial clearance in human volunteers challenged with Helicobacter pylori revealed by randomised controlled vaccination with Ty21a-based Salmonella vaccines.

BACKGROUND: Helicobacter pylori remains a global health hazard, and vaccination would be ideal for its control. Natural infection appears not to induce protective immunity. Thus, the feasibility of a vaccine for humans is doubtful. METHODS: In two prospective, randomised, double-blind, controlled studies (Paul Ehrlich Institute application nos 0802/02 and 1097/01), live vaccines against H pylori were tested in human volunteers seronegative for, and without evidence of, active H pylori infection. Volunteers (n = 58) were immunised orally with Salmonella enterica serovar Typhi Ty21a expressing H pylori urease or HP0231, or solely with Ty21a, and then challenged with 2x10(5) cagPAI(-) H pylori. Adverse events, infection, humoral, cellular and mucosal immune response were monitored. Gastric biopsies were taken before and after vaccination, and postchallenge. Infection was terminated with antibiotics. RESULTS: Vaccines were well tolerated. Challenge infection induced transient, mild to moderate dyspeptic symptoms, and histological and transcriptional changes in the mucosa known from chronic infection. Vaccines did not show satisfactory protection. However, 13 of 58 volunteers, 8 vaccinees and 5 controls, became breath test negative and either cleared H pylori (5/13) completely or reduced the H pylori burden (8/13). H pylori-specific T helper cells were detected in 9 of these 13 (69%), but only in 6 of 45 (13%) breath test-positive volunteers (p = 0.0002; Fisher exact test). T cells were either vaccine induced or pre-existing, depending on the volunteer. CONCLUSION: Challenge infection offers a controlled model for vaccine testing. Importantly, it revealed evidence for T cell-mediated immunity against H pylori infection in humans.

Dissection of the CMV specific T-cell response is required for optimized cardiac transplant monitoring.

Despite the success of antivirals in preventing clinically overt CMV disease in cardiac allograft recipients, sub-clinical active CMV infection remains a major concern because of its association with allograft rejection and vasculopathy. The measurement of CMV specific T-cell responses is a promising approach to assessing this situation. For simplicity, class-I MHC/peptide-multimers staining CD8 T-cells directly are often used but this ignores a much wider range of responses including the whole CD4 T-cell compartment. CD4 T-cells, however, were recently shown to be critical to reducing CMV load early after transplantation. To determine how extensive T-cell responses to CMV are, the responses to two dominant CMV proteins, IE-1 and pp65, were dissected in detail accounting for T-cell lineage, frequencies, epitope recognition and changes over time in more than 25 heart transplant recipients. Cross-sectional results from over 30 healthy CMV-carriers were analyzed for comparison. Responses were unexpectedly complex, with considerable inter-individual variation in terms of dominance, breadth, and recognized epitopes. Whereas the use of MHC/peptide-multimers for clinical CD8 T-cell response monitoring alone can be justified in some situations, short term T-cell activation combined with intracellular cytokine staining was clearly found to be of more general usefulness. The performance of IFN-gamma, TNF-alpha, or IL-2 as single read-outs in identifying activated T-cells was examined and confirmed that the frequently used IFN-gamma was best suited. These results should be used to inform the design of clinically applicable and diagnostically useful approaches to monitoring CMV specific responses in heart transplant recipients.

[Effect of xymedone on high-voltage-activated Ca2+ currents in pyramidal neurons of the entorhinal cortex].

The effect of xymedon on Ca2+ currents in entorhinal cortex LIII pyramidal neurons was studied using brain slices from 10-17-day old rats, which were analyzed by means of the infrared video assisted whole cell patch clamp recording. The sample slices were superfused with artificial cerebrospinal fluid containing tetrodotoxin, 4-aminopyridine, and tetraethylammonium for the blocking of Na+ and K+ channels, respectively. Xymedone was added to artificial cerebrospinal fluid and to all extracellular solutions. The slices were exposed to different concentrations of xymedone for 3 hours followed by patch-clamp recordings. Control recordings were run with the vehicle. Xymedone in a concentration of 0.01 mM decreased the maximum voltage-dependent Ca2+ current amplitude by 39.8 %, while 1 mM of xymedone inhibited the Ca2+ currents almost completely. The obtained data showed for the first time that xymedone exhibits a calcium channel blocker activity in neurons. Possible neuroprotective mechanisms of xymedone are discussed.

[Induction of apoptosis of tumor cells by binase].

An induction of apoptosis by RNase from Bacillus intermedius (binase) and its mutants characterized with low catalytic activity (Lys26Ala and His101Glu) in human myelogenic erythroleukemia K562 cells, human lung carcinoma A549 cells and human peripheral blood mononuclear cells was studied. For the first time selective apoptogenic effects of binase toward leukemic blood cells was determined. Neither antiproliferative nor apoptotic effects of binase were detected in normal human peripheral blood mononuclear cells. Formation of low molecular weight oligonucleosomal DNA fragments (less than 50 Kb) which are an early marks of apoptosis was registered in solid tumor cells treated by binase. Using mutant RNases it was shown that decrease of catalytic activity to 2.5% of wild type enzyme activity leads to the loss of apoptogenic properties of enzyme. Selective apoptogenicity of binase found towards malignant cells confirmed that antitumor agents based on bacterial RNases could be considered as an alternative to standard chemotherapeutic drugs.

[Apoptosis in the nervous system]. / Apoptoz v nervnoi sisteme.

Apoptosis (from Greek apoptosis, i.e., falling of leaves) is the phenomenon of programmed cell death, which plays an important role in the normal embryonic development and maintenance of homeostasis of the differentiated tissues of adult organisms. Completion of the apoptosis process is accompanied by specific morphological and biochemical changes in the involved cells. Various disturbances in the control of apoptosis underlie various neurodegenerative diseases, the formation of malignant tumors, autoimmune disturbances, and developmental abnormalities. A deficit of neurotrophic factors leads to apoptosis of neurons. Survival of specific cell populations of neurons is controlled by neurotrophic factors and their combinations. Oncogene bcl-2, a repressor of cell death, belongs to the better studied factors controlling apoptosis. The terminal stages of cell death, including death of neurons, depend on the activation of caspases, specifically caspase-1 (interleukin-1 beta-converting enzyme). Ca2+ and reactive forms of oxygen play an important role in the initiation of apoptosis by changing mitochondrial permeability. Neuregulin, a factor of neuronal origin, is the main controlling factor in apoptosis of Schwann cells, and this process determines the size of their definitive population. Fibroblast growth factor b diminishes apoptosis of Schwann cells in regenerating nerve fibers.

[Xymedon decreases phosphatidylserine membrane expression induced by proapoptogenic deficit of serum growth factors in Jurkat T-cells]. / Ksimedon snizhaet membrannuiu ékspressiiu fosfotidilserina, vyzvannuiu proapoptogennym defitsitom syvorotochnykh faktorov rosta v T-kletkakh Jurkat.

The effect of xymedone, a non-glucoside analog of pyridine nucleosides, on the apoptosis of human CD4+ T cells of the Jurkat line was studied by laser flow cytometry method. Xymedone decreased the membrane expression of phosphatidylserine and suppressed the increase in permeability of the cytoplasmic membrane, thus inhibiting the onset of a degradation stage of the apoptotic cascade. Possible mechanisms of the antiapoptogenic effect of xymedone within the framework of a (cytochrome C/caspase 3)-dependent signal pathway of the apoptosis are discussed.

[Potential role of the antimutagenic effect of xymedone in modification of immunoreactivity]. / Potentsialnaia rol' antimutagennogo éffekta preparata ksimedona v modifokatsii immunoreaktivnosti.

Effects of the pyrimidine derivatives of xymedone and methyluracil upon the induction of point mutations of the base pair substitution type in the Salmonella/microsome test and the frequency of chromosome aberrations in the lymphocytes of patients with the chronic osteomyelitis diagnosis. Xymedone more effectively than methyluracil inhibited the induction of point mutations by nitrosomethylurea. In the case of the cyclophosphane induced mutagenesis, the two preparations exhibited comparable antimutagen effects. Both xymedone and methyluracil (1.5 g/day, 10 days) reduced the (increased) frequency of chromosomal aberrations in the lymphocytes of patients with the chronic osteomyelitis diagnosis. The anticlastogenic effect of methyluracil vanished 5 days after termination of the 10-day administration course, while xymedone exhibited an afteraction anticlastogenic effect over this period. Interrelation of the antimutagen effect and immunomodulating activity of xymedone is discussed.

[Effect of pyrimidine derivatives on the regulation of active transport of calcium in the immunocompetent cells]. / Vliianie pirimidinovykh proizvodnykh na sistemu reguliatsii aktivnogo transporta kal'tsiia v immunokompetentnykh kletkakh.

Experiments with guinea pig immunocompetent cells and nonadherent and adherent mononuclear fractions of the peripheral blood of healthy donors, and patients with rheumatoid arthritis and purulent surgical infection disclosed different levels of Ca(2+)-ATPase activity. Thymectomy in adult guinea pigs led to diminution of activity of the enzyme in all lymphoid organs. The effect of ximedon (30 mg/kg) on Ca(2+)-ATPase activity depended on the thymus and duration of treatment with the drug, and was of a two-phase character.

[The effect of pyrimidine derivatives on the sulfhydryl status of immunocompetent cells in vivo: the interrelationship with Ca2(+)-ATPase and antimutagenic activity]. / Vliianie pirimidinovykh proizvodnykh na sul'gidril'nyi status immunokompetentnykh kletok in vivo: vzaimosviaz' s Ca2(+)-ATFaznoi i antimutagennoi aktivnost'iu.

It was demonstrated in experiments on guinea pigs that a 30 mg/kg dose of ximedon changes the content of SH-groups in immunocompetent cells depending on the duration of drug administration and the organ to which the cells under test belong. Maximum increase of the SH-groups was recorded in the thymocytes. The dynamics of changes in Ca(2+)-ATPase activity repeated the fluctuations in the SH-group content in the splenic and bone marrow cells. In the bone marrow cells of CBA x C57B1 mice 3 mg/kg ximedon suppressed the induction of chromosome aberrations caused by cyclophosphane. The interrelationship of changes in the cell sulfhydryl status with Ca(2+)-ATPase activity and the antimutagenic effect of ximedon is discussed.

[Mechanisms of pyrimidine derivatives systemic immunomodulating effect]. / Mechanizmy sistemnogo immunomoduliruiushchego delstviia pirimidinovykh proizvodnykh.

Experiments on guinea pigs showed that xymedon in a dose of 30 mg/kg stimulates nucleic acid synthesis in thymocytes and increases the blood serum nucleic acid content. Xymedon increased secretion of endogenous DNA by human lymphocytes stimulated with PNA in vitro. The systemic immunotropic mechanisms of pyrimidine derivatives are discussed.

[The immunomodulator ximedon lowers the level of induced DNA damages in bone marrow and peripheral blood cells: the possibilities for immunogenetic correction]. / Immunomoduliator ksimedon snizhaet uroven' indutsirovannykh povrezhdenii DNK v kletkakh kostnogo mozga i perifericheskoi krovi: vozmozhnosti immunogeneticheskoi korrektsii.

It was established in experiments on albino unbred mice and during treatment of patients with osteomyelitis that 30 mg/kg of ximidon suppresses the formation of micronuclear polychromatophilic erythrocytes found in the bone marrow of mice and peripheral blood of patients with chronic osteomyelitis. The interrelationship of the results obtained with the modulating effect of ximidon on the mitochondrial, thiol, and adenylate cyclase-dependent mechanisms of cell regulation is discussed.

[Assessing the toxic effect of 2,4,6-trinitrotoluene on cells of escherichia coli K12 by flow cytofluorometry].

The magnitude of transmembrane potential delta psi in cells of Escherichia coli K12 was determined by the method of flow cytofluorometry for different phases of growth. It was large in the log phase, whereas in the lag and stationary phases, the population was shown to consist of two subpopulations with low and large values of delta psi in cells. In the presence of 2,4,6-trinitrotoluene (TNT), this bimodal distribution of delta psi over the population was observed during the entire growth period until TNT was almost completely eliminated from the cultivation medium (to a concentration of 18-20 mg/l). The mean value of delta psi in cells of the population grown in the presence of TNT was substantially smaller than that in controls due to the larger size of the subpopulation with a low value of delta psi. Upon elimination of TNT, the distribution of delta psi in cells of the culture became unimodal and close to that in the control culture in the early log phase of growth. These findings are discussed from the standpoint that considers heterogeneity of the culture of Escherichia coli K12 as a mechanism of its adaptation to the presence of xenobiotics.

[Use of systemic immunomodulator xymedone in distructive pulmonary tuberculosis]. / Primeneniie sistemnogo immunomoduliatora ksimedona pri destruktivnykh formakh tuberkuleza legkikh.

The study was undertaken to examine 59 patients with destructive pulmonary tuberculosis who had T-cell dysfunction (loss of CD3+, CD4+, lymphocytes, slightly positive tuberculin test) and augmented IgA and IgG production. Oral Xymedone given in a dose 2.0 g daily for two 2 months in combination with antibacterial therapy abolished lymphopenia, restored CD4+ counts, CD4+/CD3+ ratio, upregulated IgG levels, but did not affect IgA levels. The Xymedone-treated patients developed fewer side effects due to basic antibacterial chemotherapy and showed more rapid culture conversion, resolution of pulmonary infiltration and closure of destructive cavities.