The Level of Anti-Polyethylene Glycol (PEG) Immunoglobulin G Antibodies in Human Serum Does Not Correlate with Efficiency of PEGylated Nanoparticle Uptake by Monocytes.

The presence of anti-polyethylene glycol (PEG) antibodies can limit the clinical efficacy of PEGylated drugs and cause anaphylactic reactions in patients. Monocytes/macrophages are effector cells involved in IgG-mediated passive systemic anaphylaxis. We studied the influence of human blood serum on the efficiency of uptake of PEGylated nanoparticles by human blood monocytes. It has been shown that magnetic nanoparticles modified with PEG-3000 and solid lipid nanoparticles containing PEG-2000 are avidly internalized by human blood monocytes in vitro, the uptake efficiency depends on the features (composition) of donor blood serum, but does not correlate with the level of the IgG antibody against PEG.

Insulin Resistance in Experimental Type 1 Diabetes Mellitus.

Experimental type 1 diabetes mellitus (T1DM) was induced in rats by daily intraperitoneal injections of alloxan in a dose of 90 mg/kg for 4 days. For verification of insulin resistance, insulin tolerance test was performed in 2 weeks and the glucose utilization rate constant (KITT) was calculated. The rats demonstrated the main symptoms of T1DM: hypoinsulinemia, hyperglycemia, ketonemia, glucosuria, ketonuria, polydipsia, polyphagia, weight loss, and insulin resistance, as evidenced by a decrease in KITT. The serum content of free fatty acids and triacylglycerols significantly increased. The content of triacylglycerols increased in skeletal muscles and decreased in the liver. A negative linear correlation was found between KITT and triacylglycerol content in muscles. Thus, the development of insulin resistance in experimental T1DM in rats is associated with accumulation of triacylglycerols in skeletal muscles.

Comparative Evaluation of ß-Catenin and E-Cadherin Expression in Liquid Aspiration Biopsy Specimens of Thyroid Nodules.

We compared the results of gene molecular and immunocytochemical studies of ß-catenin and E-cadherin in different variants of nodular thyroid disease (nodular colloid goiter, follicular thyroid adenocarcinoma, papillary thyroid cancer) and revealed changes of the function of the E-cadherin/ß-catenin complex leading to switching from adhesion function of ß-catenin in nodular colloid goiter to predominantly transcriptional activity in papillary carcinoma. The results confirm the important role of disturbances in E-cadherin-ß-catenin interactions in the mechanisms of malignant transformation of follicular epithelium.

[The technique of fluid cytology in diagnostic of thyroid diseases].

The increasing of number of diseases of thyroid actualizes searching for new solutions in differential diagnostic. Nowadays, the technique of fluid cytology is started to be applied under cytological analysis of puncture sample of thyroid. However, experience of application of technique of fluid cytology in interpretation of cytological sample is limited. The article presents results of analysis of samples of fine needle aspiration biopsy of thyroid from 126 patients. In case of traditional cytological analysis aspirate was putted on glass right away. In case of technique of fluid cytology patient was re-punctured and sample was placed into vial with preservative solution. At re-puncture increasing number of analysis with non-diagnosed samples was observed (40.6% in comparison with 17.8% under traditional cytological analysis). However, in 50% of patients with non-informative samples in traditional cytological analysis implementation of technique of fluid cytology permitted to establish proper diagnosis. In diagnostic of non-tumor affections of thyroid the best results demonstrated by technique of fluid cytology (86.6% as compared with 47.2% under technique of fluid cytology). However, in diagnostic of benign tumor affections of thyroid the best accuracy was demonstrated by traditional cytological analysis (61.1% as compared with 33.3% of technique of fluid cytology). Both techniques demonstrated similar results in category of malignant affections of thyroid. The diagnosis of cancer or suspicion to cancer was established in 85.7% of cases under technique of fluid cytology and in 82.7% of cases under traditional cytological analysis). The analysis of cytological pictures of various diseases of thyroid in technique of fluid cytology demonstrated specifics and differences from traditional cytology that are to be considered. At the same time, observations convincingly testify that in number of cases traditional and fluid techniques supplement each other. Besides, technique of fluid cytology permits preserving and transport puncture samples and in case of necessity to apply additional clarifying diagnostic techniques that in turn increases effectiveness of cytological diagnostic of affections of thyroid.

[ß-Catenin: Structure, Function and Role in Malignant Transformation of Epithelial Cells].

The article presents the data on the structure and mechanisms of ß-catenin functioning. The basic aspects of the role of ß-catenin in malignant transformation have been studied at various tumors. Primary structure of ß-catenin allows it to interact with many factors and ligands, including transcription factors, α-catenin, cadherin, Axin, Rho family GTPases, Bcl9 et al. This interaction is the base for ß-catenin's intracellular multi-functioning. The review presents data on the participation of ß-catenin in the mechanisms of adhesion, regulation of RNA metabolism, formation contacts with the cytoskeleton and its role in the canonical Wnt signaling pathway, marked examples pro-inflammatory and anti-inflammatory effects of ß-catenin. The ß-catenin involvement in malignant transformation and progression of certain tumors is not in doubt. The data on the changes in ß-catenin expression in the given examples of colon cancer, prostate cancer, different forms of thyroid cancer and hepatocellular carcinoma are presented with the prospects of its use as a marker and a predictor of malignant transformation. Continued research in this area will not only make use of ß-catenin as a potential predictor of malignant tumors, but also to develop approaches to targeted therapy.

Role of heat shock protein 27 in regulation of glutathione system and apoptosis of Jurkat tumor cells and blood lymphocytes.

Reaction of the glutathione system of Jurkat tumor cells and blood lymphocytes was evaluated under conditions of culturing with 5-(5-ethyl-2-hydroxy-4-methoxyphenyl)-4-(4-methoxyphenyl) isoxazole (KRIBB3), a selective inhibitor of heat shock protein Hsp27. The results indicated the regulatory role of Hsp27 in the maintenance of the functional activities of glutathione reductase, glutathione peroxidase, and realization of apoptotic death of Jurkat cells and blood lymphocytes. Inhibition of Hsp27 in Jurkat tumor cells led to imbalance of the glutathione system and increase of the share of annexin-positive cells.

Effect of galectin-1 on apoptosis of CD4(+) lymphocytes differentiated in vitro towards regulatory T cells.

We studied the effect of galectin-1 on apoptosis of CD4(+) lymphocytes intact and in vitro differentiated towards regulatory T cells. An increase in the content of apoptotic CD4(+) lymphocytes was observed after exposure of intact cells with 15 ng/ml galectin-1 and after exposure of regulatory T cells with 10 and 15 ng/ml galectin-1. Apoptosis of regulatory T cells induced by galectin-1 was accompanied by an increase in the content of proapoptotic protein Bad.

[Regulation of the genes expression of CD4+ T-lymphocytes differentiation transcription factors by galectin-3 in vitro].

Now a number of CD4+ T-lymphocytes, known as Th1, Th2, Treg and Th17, is currently identified and well- studied. The methods basing on the targeted regulation of differentiation process of the Th-lymphocytes that carry out the immune response polarization attract an attention of scientists dealing with a correction of immune-mediated. In the present study, endogenous beta-galactoside-binding protein of the lectin family, galectin-3, was investigated as a regulator of T-cell homeostasis. A galectin-3 is known to be actively produced by tumor cells in malignant transformation and able to influence the processes of signal transduction, cell-cell cooperation and the implementation of programmed death. As cell differentiation processes are directly connected with the regulation of gene expression, we investigated the effect of recombinant galectin-3 on expression of mRNA of transcription.factors, which guide the differentiation of CD4+ lymphocytes. The study was performed on peripheral blood mononuclear cells of healthy individuals. The gene expression levels were evaluated by a real-time PCR. In the experiments in vitro, it has been first found the recombinant galectin-3 (0.5 mg/mL) up-regulating the expression of transcription factors Gata-3 and Rorc mRNAs and down-regulating the mRNA expression of transcription factors T-bet and FoxP3. Up to a concentration of 1 mg/mL recombinant galectin-3 stimulates Th-cells by dose-dependent manner, whereas at higher concentrations stimulating effect weakens, and inhibiting action starts prevailing. Thus, one can suppose that galectin-3 through regulation of lymphocytes differentiation promote development of allergic, autoimmune and neoplastic diseases that allows us to consider the galectin-3 as a.potential target for therapy of these diseases.

[Molecular pato-physiology of chronic virus infections].

Article is based on published data and results of own original researches on development of chronic socially inportant virus infections. Special attention is paid to priority research direction--molecular-genetic markers of chronis virus infections predisposition, development of personalized therapy molecural basis and forecast of infect and man organism interrelation results.

[Change in the production of key cytokines for regulation eosinophilic granulocytes in Opisthorchis felineus invasion].

Opisthorchis invasion is accompanied by the imbalanced lymphocytic subpopulational composition manifested itself as induction of the B-link and, on the contrary, depression of T-lymphocytic populations (CD4+, CD8+), with their weaker helper-suppressor association. The immunocompetent cells were ascertained to show a higher production of TH2 cytokines that had an eosinophil-stumulating effect.

[Alleles distribution of polymorphic promoter region C-590T in interleukin-4 genes and Q-576r and Ile-50Val regions in IL-4 receptor gene IL-4RA in patients with HCV-infection].

81 patients with confirmed HCV-infection and 48 healthy volunteers were examined. In healthy Caucasian participants living in Siberian region significant predominance of C/T genotype in promoter region C-590T of interleukin-4 (IL-4) gene and Q/Q and Ile/Val genotypes in points -50 and -576 of IL-4RA gene that codes alpha-chains of IL-4 receptor were revealed. In patients with HCV-infection predominance of C/T genotype in C-590T region in IL-4 gene (OR = 1.86), R/R genotype in Q-576R region of IL-4RA gene (OR=7.86), and Val/Val genotype in point Ile-50Val (OR = 2.6) of the same gene. Summary predictive coefficient of hepatitis C development in carriers of these genotypes approached to 95%. During analysis of role of allelic polymorphism of IL-4 genes in predisposition to hepatitis C development it is necessary to consider not only presence of allelic variants of promoter regions of the IL-4 genes, but also the polymorphism of genes coding molecules binding with this cytokine on target cells membranes and in its soluble form.

Role of genetically determined production of immunoregulatory cytokines in immunopathogenesis of chronic viral hepatitides.

Immunopathogenesis of chronic viral hepatitides was studied by modern immunological, molecular, genetic methods. We revealed an imbalance in the production of immunoregulatory cytokines by mononuclear leukocytes (primarily of the Th2 type). The risk of progression and chronic course of viral hepatitides in Caucasian population was associated with alleles of promoter regions -330G and -592A in the IL-2 and IL-10 genes, respectively, as well as with the T/T genotype of the polymorphic region C590T in the IL-4 gene. The C/C genotype of the IL-10 gene promoter region C592A was shown to be a factor determining resistance to long-term persistence of hepatitis B and C viruses.

Dysregulation of apoptotic death in the pathogenesis of virus-induced cytogenetic instability of blood lymphocytes.

The cytogenetic status and activity of regulatory systems for stability of the cell genome were evaluated in patients with chronic viral persistence. Hepatitis B and C viruses damage the chromosome apparatus of peripheral blood lymphocytes. Cytogenetic instability of immunocompetent cells during chronic viral infection was associated with inhibition of DNA excision repair system and dysregulation of apoptosis in target cells.

[Cytokines and antiviral immuniy].

In the review of modern literature and results of own research about role of immunoregulation cytokines imbalance system in pathogen of deep-rooted virus infections are analyzed. The question on molecular mechanisms of cytokine chain deregulation at persistent virus infections is discussed.

[Immunopathogenetic features of tick-borne encephalitis under long-term viral antigenemia].

The occurrence of chronic tick-borne encephalitis viral antigenemia characterized by the asymptomatic course or minimal clinical manifestations is caused by the virus-immune cell interaction. Cellular immunity was studied in 183 patients with chronic (more than 6-month) tick-borne encephalitis viral antigenemia, by evaluating the immunophenotypic and cytogenetic statuses, structural and functional features, cytokine profile, and peripheral lymphocytic apoptosis. The findings suggest impairments in the cooperation of immunocompetent cells and in the classical scheme of antiviral immunity regulation in tick-borne encephalitis viral persistence due to the changes in lymphocytic structural and functional properties.

[Dysregulation of cytokine production by mononuclear cells of peripheral blood in persistent infection with a virus of tick-borne encephalitis].

Tick-borne encephalitis is accompanied by dysregulation of cytokine production of mononuclear leucocytes of peripheral blood. The changes of production of IFN-r by mononuclear leucocytes of peripheral blood positively correlated with a degree of activity of pathological process and intensity of clinical symptoms of neuroninfection. On the contrary, secretion of IL-12 mononuclears directly correlated with virus persistence in the organism and does not depend on clinical presentation of the disease.

Molecular mechanisms of changes in TNF-alpha production by blood mononuclears in chronic viral hepatitis C.

Chronic viral hepatitis C is associated with decreased production of TNF-alpha by the peripheral blood mononuclear leukocytes irrespective of virus genotype and degree of the morphological activity of the process in the liver. This process positively correlates with the increase in the content of TNF-alpha soluble receptor (molecular weight 55 kDa), which can play a role in the mechanisms of immunopathogenesis of long persistence of hepatitis C virus in the body.