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Objective. To evaluate the ability of a combination of budesonide and azithromycin to influence the migration of blood monocytes and lymphocytes in patients with chronic obstructive pulmonary disease (COPD).Materials and methods. Peripheral blood mononuclear cells from patients with COPD (n=8) were incubated with glucocorticoid budesonide (10 nM), macrolide antibiotic azithromycin (10 µg/mL), or their combination, and then transferred to chemotaxis chambers containing chemokines RANTES (CCL5, 10 nM) or IP-10 (CXCL10, 10 nM). Cells migrated to the lower compartment of the chamber were collected, stained with monoclonal antibodies to CD3, CD14, CD19, CD45 and counted on a flow cytometer.Results. Azithromycin alone and in combination with budesonide inhibited the migration of blood T-lymphocytes and B-cells and enhanced the migration of blood monocytes to RANTES and IP-10. The combination of azithromycin and budesonide had a more suppressive effect on the chemotaxis of blood T- and B-lymphocytes to RANTES and IP-10 than budesonide alone. The combination of azithromycin and budesonide had an effect similar to azithromycin alone on the migration of blood T- and B-lymphocytes, as well as monocytes in patients with COPD.Conclusion. The results of the study demonstrate the ability of azithromycin alone to modulate the chemotaxis of peripheral blood monocytes and lymphocytes in patients with COPD and the lack of advantages of its combination with budesonide.

Chronic obstructive pulmonary disease (COPD) is characterized by reduced sensitivity of cells to the anti-inflammatory effects of glucocorticoids (GCs). Azithromycin and a low dose theophylline have a significant impact on molecular mechanisms leading to corticosteroid resistance. The aim of this study was to evaluate the ability of azithromycin and theophylline to enhance the anti-inflammatory effects of GCs on the production of cytokines by NK and NKT-like blood cells of COPD patients. Whole blood cells from COPD patients (n=21) were incubated in the presence of budesonide (10 nM), azithromycin (10 μg/mL), theophylline (1 μM), or their combinations and stimulated with phorbol myristate acetate (50 ng/mL). Intracellular production of proinflammatory cytokines in NK (CD3-CD56+) and NKT-like (CD3+CD56+) blood cells was analyzed by flow cytometry. Budesonide reduced synthesis of interleukin 4 (IL-4), CXCL8, tumor necrosis factor α (TNFα) by NK and NKT-like cells, as well as production of interferon γ (IFNγ) by NK cells. Azithromycin suppressed production of IL-4 and CXCL8 by NK and NKT-like cells, and theophylline inhibited IL-4 synthesis by these lymphocytes. The combination of azithromycin and budesonide had a more pronounced inhibitory effect on the production of IL-4 and CXCL8 by NK and NKT-like cells than the effect of these drugs alone. The combination of theophylline and budesonide suppressed synthesis of IL-4 and CXCL8 by NK and NKT-like cells, as well as the production of TNFα and IFNγ by NK cells stronger than budesonide alone. The obtained results demonstrate the benefits for the combined use of GCs with theophylline at a low dose or azithromycin to suppress the inflammatory process in patients with COPD.Khronicheskaia obstruktivnaia bolezn' legkikh (KhOBL) kharakterizuetsia snizhennoĭ chuvstvitel'nost'iu kletok k protivovospalitel'nym éffektam gliukokortikosteroidov (GKS). Izvestno, chto teofillin v nizkoĭ kontsentratsii i azitromitsin mogut vliiat' na molekuliarnye mekhanizmy razvitiia steroidorezistentnosti. Tsel'iu nastoiashchego issledovaniia byla otsenka sposobnosti azitromitsina i teofillina usilivat' protivovospalitel'noe deĭstvie GKS na produktsiiu tsitokinov NK i NKT-podobnymi kletkami krovi patsientov s KhOBL. Kletki tsel'noĭ krovi patsientov c KhOBL (n=21) inkubirovali v prisutstvii budesonida (10 nM), azitromitsina (10 mkg/ml), teofillina (1 mkM) ili ikh kombinatsiĭ i stimulirovali forbol-miristat-atsetatom (50 ng/ml). Vnutrikletochnuiu produktsiiu provospalitel'nykh tsitokinov v NK (CD3-CD56+) i NKT-podobnykh (CD3+CD56+) kletkakh krovi analizirovali metodom protochnoĭ tsitometrii. Budesonid snizhal sintez interleĭkina 4 (IL-4), CXCL8, faktora nekroza opukholi α (FNOα) NK i NKT-podobnymi kletkami, a takzhe produktsiiu interferona γ (IFNγ) NK kletkami. Azitromitsin podavlial sintez IL-4 i CXCL8 NK i NKT-podobnymi kletkami, a teofillin ingibiroval produktsiiu IL-4 étimi limfotsitami. Sochetanie azitromitsina i budesonida okazyvalo bolee vyrazhennoe ingibiruiushchee vozdeĭstvie na produktsiiu IL-4 i CXCL8 NK i NKT-podobnymi kletkami, chem deĭstvie liubogo iz étikh preparatov. Kombinatsiia teofillina i budesonida supressirovala sintez IL-4 i CXCL8 NK i NKT-podobnymi kletkami, a takzhe produktsiiu FNOα i IFNγ NK kletkami bolee znachitel'no, chem odin budesonid. Poluchennye dannye demonstriruiut preimushchestva kombinirovannogo ispol'zovaniia GKS sovmestno s teofillinom v nizkoĭ kontsentratsii libo azitromitsinom dlia podavleniia vospalitel'nogo protsessa u patsientov s KhOBL.

4-Methyl-7-(4-methyl-2-oxo-2H-chromen-6-yl)disulfanyl-2H-chromen-2-one hydrotriiodide was synthesized by the reaction of 7-mercapto-4-methylcoumarin with iodine, and its structure was determined by NMR spectroscopy and X-ray diffraction combined with Raman and IR spectroscopy. The molecular and crystal structure of the reaction product was determined.

The crystal and molecular structure of the reaction product of imidazolidine-2-thione and molecular iodine of the composition (C3H6N2S)2•3I2 was established by X-ray diffraction. The crystal structure is composed of strongly connected five-component associates consisting of three I2 molecules and two heterocyclic molecules. Quantum chemical calculations followed by the topological analysis of the electron density function provided an estimate of the strongest intermolecular interactions in the crystal structure of the molecular adduct.

In the present study we have analyzed superinduction of TNF-alpha mRNA and enhancement of TNF-alpha gene transcription by cycloheximide (Chx) in human blood monocytes isolated by continuous Percoll gradient and activated in vitro. In the same monocyte cultures, we have compared the rate of gene transcription of TNF-alpha, IL-1 beta, IL-8, and the P53-antioncogene under the influence of plastic adherence, Staphylococcus aureus Cowan 1 (SAC), and Chx added at different times of monocyte culture. It was shown that the cytokine genes have low or negligible transcriptional activity in freshly isolated monocytes, whereas P53 gene transcription was constant in freshly isolated and in vitro-stimulated cells. Transcription of the IL-1 beta and IL-8 genes was induced by adherence and was not more enhanced by SAC. Transcription of the TNF-alpha gene was not induced by adherence. Chx added at the beginning of the monocyte culture did not block TNF-alpha or IL-1 beta gene transcription. IL-8 gene transcription, however, was abrogated by Chx. Addition of SAC to monocyte culture containing Chx caused significant enhancement of TNF-alpha gene transcription. Addition of Chx after 2.5 or 4 h of SAC activation caused "superinduction" of TNF-alpha mRNA and enhancement of TNF-alpha gene transcription. The data imply that TNF-alpha gene transcription in activated human monocytes might be regulated by both positive and negative regulatory factors that differ in their stability and protein synthesis dependence. In addition, results demonstrate that TNF-alpha, IL-1 beta, IL-8, and p53 genes in human monocytes are differently regulated.

The p53 gene has been associated with malignant transformation as well as "anti-oncogene" activity. In the present report expression of p53 in resting and activated human blood monocytes and lymphocytes is analyzed. It is found that human monocytes freshly isolated by continuous percoll gradient centrifugation contained detectable level of p53 mRNA. Stimulation of monocytes by potent activation inducer Staphylococcus Aureus Cowan I for 3-5 hr caused disappearance of r53 mRNA. In contrast, induction of high level of TNF-alpha mRNA was detected. Addition of cycloheximide had no effect on p53 mRNA content in stimulated monocytes, and caused disappearance of mRNA in resting cells. In lymphocytes cultures p53 mRNA was absent in freshly isolated cells and in resting lymphocytes cultured for 20 hr. Activation of lymphocytes by lectin caused accumulation of p53 mRNA. We suggest that r53 gene regulation and functions might be different in human monocytes and lymphocytes.

The addition of recombinant tumor necrosis factor or monoclonal antibodies to this factor into human monocyte culture does not change the synthesis of interleukin-8. By contrast, tumor necrosis factor induces the synthesis of interleukin-8 in human lymphocytes.

Defensins, a family of small, cationic, antimicrobial peptides, are found in mammals, insects and plants. alpha-defensins are stored in granules of neutrophils and released upon activation by exocytosis. It was shown here that human neutrophil peptide (HNP), at concentrations of 10(-8) -10(-9) M, up-regulated the expression of TNF-alpha and IL-1 beta in monocytes activated with Staphylococcus aureus or PMA, while expression of IL-10 mRNA was down-regulated and production of IL-8 was not affected. HNP alone was unable to induce TNF-alpha or IL-1 beta expression in resting monocytes. At concentrations of 10(-4) -10(-5)M, HNP was cytotoxic for monocytes in serum-free medium. The cytotoxicity was abrogated in the presence of serum, while a cytokine-modulating effect of HNP was observed in the presence of serum and in whole blood, suggesting that this mechanism may function in vivo. Similarly, serum did not abrogate bactericidal activity of HNP. It was also demonstrated herein that HNP at 10 (-8) -10(-9) M, attenuated the inhibitory action of dexamethasone on TNF-alpha production. In parallel to monocyte studies, we have showed that HNP at concentrations ranging from 10(-9)M to 10(-6)M caused about 5-fold suppression of VCAM-1 expression in TNF-alpha-activated human umbilical vein endothelial cells, while the ICAM-1 expression was not affected. Our findings suggest that neutrophil defensins have the potential to modulate the inflammatory responses through regulation of cytokine production and adhesion molecule expression.

The influence of human defensin HNP-1 on the synthesis of tumor necrosis factor-alpha (TNF-alpha) by human blood monocytes was studied. It was shown that TNF-alpha production by human monocytes activated by SAC or PMA was augmented in the presence of HNP-1 concentrations of 10(-8)-10(-9) M. HNP-1 alone induced no synthesis of TNF-alpha. High concentration of HNP-1 (10(-4) M) was cytotoxic for human monocytes.

Antiviral factor (AF) of protein nature has been isolated from chick embryo fibroblasts infected with Venezuelan equine encephalitis virus. The suppression of virus reproduction has been observed both in homologous and heterologous cell cultures when the preparation was introduced immediately after the adsorption of the virus after pretreatment of the cell monolayer. The study has demonstrated that the antiviral effect of AF is not linked with its IFN-alpha and TNF-alpha activity. Analysis of the results obtained in this study and earlier data contained in literature suggests that infected chick embryo fibroblasts release original cytokine of non-interferon nature with antiviral activity.