[Prospects for the development of new prolonged forms of valproic acid derivatives for the relief of convulsive syndrome]. / Perspektivy razrabotki novykh prolongirovannykh form proizvodnykh val'proevoi kisloty dlya kupirovaniya sudorozhnogo sindroma.

Poisoning with pesticides containing convulsive substances is the cause of death of thousand people in the world. Convulsive syndrome can also develop with infection of the central nervous system, intracranial hemorrhages and strokes, alcohol withdrawal syndrome, eclampsia. Up to 10% of all of the people on the Earth had experienced seizures. The most common cause of seizures is epilepsy. Existing drugs are not always effective and have side effects. Alternative therapies are being developed. Valproates (derivatives of valproic acid) are one of the most studied drugs of choice for epilepsy and other variants of convulsive syndrome, have many years of experience in use, are included in international and Russian clinical guidelines for the treatment of epilepsy. The antiepileptic, mood stabilizing and neuroprotective properties of valproate due to pre- and postsynaptic modulation of GABA-ergic transmission, regulation of glutamate activity, serotonin and dopamine levels in the hippocampus, modulation of sodium, calcium and potassium channels, epigenetic effects on the body are described. However, valproates have a number of undesirable side effects, usually associated with high doses of drugs used for a long time. Attempts to reduce the risk through the development of drugs with controlled release of the active substance are not always effective - the high individual sensitivity of about 15% of patients with epilepsy makes standard doses toxic, the pharmacokinetics and effectiveness of the developed generics are significantly inferior to the original imported drug. An alternative could be the development of Russian preparations containing a proactive component from which valproic acid is gradually released during the metabolism.

[Immunoregulatory proteins and cytokines in the blood of patients with Graves' disease].

BACKGROUND: Immunoregulatory proteins (alpha-2-macroglobulin, lactoferrin) actively participate in inflammatory and autoimmune processes, affect synthesis and transport of hormones and cytokines, and control cell proliferation and apoptosis. However, the role of these proteins in the pathogenesis of Graves' disease (GD) is poorly understood. Objective - the study objective was to determine blood levels of alpha-2-macroglobulin (α2-MG), lactoferrin (LF), and cytokines (TNF-α, IL-6, IL-8, and IFN-γ) in GD. MATERIAL AND METHODS: We determined blood levels of TSH, free T4, TSH receptor antibodies, TNF-α, IL-6, IL-8, IFN-γ, and LF by ELISA as well as α2-MG by quantitative rocket immunoelectrophoresis in 50 patients with decompensated and compensated (4-6 months and 1.5-2 years after treatment onset) GD and 25 healthy females (control group). RESULTS: GD clinically manifested by body weight l in 84% of patients, sinus tachycardia in almost all patients, paroxysmal atrial fibrillation in 18% of cases, endocrine ophthalmopathy in 12% of patients, and neurological changes. In decompensated GD, there was a statistically significant increase in levels of IFN-γ, IL-6, and α2-MG and an especially significant increase in levels of IL-8 and LF. At 4-6 months after treatment onset, clinical manifestations were stopped in all patients, levels of IL-6 and α2-MG decreased, but the concentrations of TSH receptor antibodies (TSHR-Abs), IL-8, IFN-γ, and LF remained elevated. At 1.5-2 years, levels of the studied proteins and cytokines did not differ from those in the control group. CONCLUSION: An increase in blood levels of IL-8, LF, IL-6, and α2-MG in incident or recurrent GD and a decrease in the levels during treatment confirm involvement of immunoregulatory proteins in pathogenesis of the disease.

[The regulatory transport proteins in diagnostic and prognosis of preeclampsia].

The article considers the results of analysis of content of regulative transport proteins in blood serum and urine of pregnant women (term III) in case of uncomplicated pregnancy and pregnancy complicated by preeclampsia and eclampsia for elaborating their pathogenic role and evaluating prognostic significance. It is established that the more severe eclampsia is the higher is the level of a2-macroglobulin and the lower is the content of lactoferrin in blood. At that, excretion of a2-macroglobulin and albumin with urine increases with aggravation of severity of processes and in urine is detected a1-antitrypsin previously undetected. The excretion of lactoferrin reaches its peak values in case of preeclampsia and decreases in case of eclampsia. The alteration of levels of a2-macroglobulin and lactoferrin are uncharacteristic for classic inflammatory reaction and testify their active involvement into pathogenesis of eclampsia. The decreasing of in blood of levels of a1-antitrypsin lesser than 5 g/l and lactoferrin lesser than 0.8 mg/l at concentration of a2-macroglobulin higher than 3.5 g/l against the background of decreased levels of albumin and crude protein in blood and also increasing in urine of concentrations of a2-macroglobulin up to 0.0005 g/l and higher and occurrence of a1-antitrypsin and increasing of content of albumin up to 10 times can be recommended as criteria of high risk of development of eclampsia in regnant women with moderately expressed preeclampsia in term III.

[The influence of local and combined acute suppurative Highmore maxillary sinusitis on the serum lactoferrin and interleukin-8 levels in the children].

The objective of the present study was to estimate the influence of isolated and combined acute suppurative Highmore maxillary sinusitis on the serum lactoferrin (LF) and interleukin-8 (IL-8) levels in the children. A total of 70 children at the age varying from 4 to 15 years were available for the examination. Twenty of them constituted the control group, 29 presented with acute suppurative Highmore maxillary sinusitis, in 21 cildren this condition was combined with frontitis, ethmoiditis, otitis, and adenoiditis. Serum lactoferrin and interleukin-8 levels were measured by solid phase enzyme-linked immunoassay. It was shown that all the aforementioned forms of rhinosinusitis were associated with a significant increase of the serum LF level, an universal factor inactivating the propagation of bacterial, viral, and fungal pathogens. The level of IL-8 known to activate chemotaxis was increased only in the children presenting with combined forms of suppurative Highmore maxillary sinusitis. It is concluded that this difference can be used for the purpose of dufferential diagnostics of different forms of suppurative maxillary sinusitis.

[Protein components of innate immunity in protection from pathogenic invasion (literature review)].

System of innate immunity is a set of effector molecules and cells counteracting invasion of pathogens and their products. Alpha-2-macroglobulin (a2-MG) and lactoferrin (LF) play a significant role in primary protection of organism. A wide spectrum of transport and regulatory functions, high affinity to endocytosis receptor as well as structural features of these proteins allow them not only to effectively protect the organism during direct contact with pathogen but also to have an immunomodulatory impact on immunocompetent cells of adaptive immunity. However despite a common receptor and a number of ligands, mechanisms of realization of protective functions ofa2-MG and LF differ significantly. The aim of this review is to systemize knowledge on means and mechanisms of protection of a 2-MG and LF against invasion.

[The proteins of fast phase of inflammation in prognosis of condition of newborn in case of pregnancy complicated by hydramnion and under the risk of intrauterine infection].

The content of alpha-macroglobulin associated with pregnancy, alpha2-glycoprotein, alpha1-antitripsin, and lactolerrin in blood serum of pregnant women and umbilical serum under hydramnion and risk of development of intrauterine infection of fetus is investigated. It is demonstrated that in case ofpresence in blood of pregnant woman of G-antibodies to C. trachomatis under low titers (1:20, 1:40) the increase of levels of alpha-macroglobulin, alpha2-glycoprotein, al-antitripsin and especially of lactoferrin in serum of pregnant women testifies the high risk of presence of intrauterine infection of fetus and probability of birth of child with low values on Apgar scale.

[The concentrations of certain cytokins and lactoferrin in blood and supernatants of cell cultures in patients with autoimmune thyroiditis].

The analysis was applied to serum and supernatants of day cultures of blood cells of patients with autoimmune thyroiditis and healthy women to determine the content of cytokins (IFN-gamma, TNF-alpha, IL-beta, IL-6, IL-10, L-17) and lactoferrin. The test-system "Cytokin-Stimul-Best" was applied. It is established that in patients with autoimmune thyroiditis the proliferative response of cells to mitogen stimulation is lowered in vitro: the levels of alpha-tumor necrosis factor interleikin-6, lactoferrin. The stimulation index of alpha-tumor necrosis factor, interleikin-6, 1beta, 10 and interferon was statistically reliably lower in supernatants of mitogen stimulated cultures of cells of patients with autoimmune thyroiditis as compared with indicators of healthy women.

[The reactants of acute phase of inflammation and anti-inflammatory cytokines under various complications of cardiac infarction].

The article presents the results of analysis of blood serum from 60 patients (aged 58.9 +/- 1.5 years) with Q-genous complicated and non-complicated cardiac infarction. The study evaluated the content of various proteins of acute phase (alpha-2-macrogloblin, alpha-1-antitripsin. lactofferin) and cytokines inducing their synthesis (IL-6, IL-1beta, L-8). The examinations carried out on 1st, 7th and 14th day of development of cardiac infarction. It is demonstrated that under cardiac infarction complicated by cardiogenic shock on first day of diseases high levels of lactoferrin and lower levels of macroglobulin are detected in blood serum. On the 1-7th day of disease the high levels of lactofferin on the background of invariable concentration of macroglobulin accompany the edema and congestive processes in lungs under cardiac infarction. On the 1-7th day the high levels of lL-6 and IL-8 were detected under large-focal cardiac infarction independently of presence/absence of complications.

[Macroglobulin family proteins in breast cancer].

Determined serum levels of alpha-2-macroglobulin (a2-MG) and pregnancy-associate alpha-2-glycoprotein (a2-PAG) in 98 breast cancer patients stage I-IV prior to the special antitumor treatment. Statistically significant negative correlative dependence of the a2-PAG in the blood and lifetime of patients is proved. We believe that the definition of pre-treatment level of a serum a2-MG and a2-PAG concentrations for patients with breast cancer, can be used as an additional predictor of life expectancy.

[Macroglobulin signaling system].

This review will focus on the systematization of knowledge about structure of macroglobulin signaling system, which includes macroglobulin family proteins (alpha-2-macroglobulin, alpha-2-glycoprotein, pregnancy associated plasma protein A), their receptors (LRP, grp78), ligands (proteinases, cytokines, hormones, lipids, et al.) transforming and transcriptional factors for regulation of macroglobulins synthesis. After reviewing the functions of macroglobulin signaling system, and mechanisms of their realization, we discuss the complex and significant role of this system in different physiological and pathological processes.

[Peculiaries of interactions between proteins of the macroglobulin family and with the endocytic receptors (a possible mechanism of transmembrane transfer)].

We have conducted a series of experiments, for specification of mechanisms which proteins of the macroglobulin family deliver regulatory substances inside of a cells. We have shown that all members of the family are not only compete for binding to proteinases, but also can interact with each other. We have confirmed that only a complex of alpha-2-macroglobulin (alpha2-MG) with proteinase is capable to react with the major endocytic receptor (low-density lipoprotein receptor-related protein, LRP). For the first time we have demonstrated, that interaction of alpha2-MG firstly with proteinase, and then with LRP provokes a progressive conformational consolidation of the multicomplex, which is accompanied by a paradoxical increase of the electrophoretic mobility in comparison with native alpha2-MG. We suggest that such stepwise conformational consolidation, together with earlier demonstrated charge neutralization (versus pI of internal environments) after interaction firstly with proteinase, and then with LRP, components of is the key moment of the mechanism of transmembrane transfer. Taking into account, that alpha2-MG transfers a broad spectrum of protein regulators, and interacts not only with LRP, but also with a signal receptor (grp78), and also can regulate (under certain conditions) both own synthesis, and synthesis of LRP and its blocker (receptor - associated protein, RAP), we suggest that this main member of the macroglobulin family plays a leading role in the regulation of intercellular interactions and in the transmission of signal inside of a cell.

[Various acute phase reactants in different types of proliferative diseases of the uterine appendages].

The authors examined serum in patients with ovarian cancer (OC; a disseminated process), ovarian cystadenoma (OCA), or external endometriosis (EM) before treatment and in apparently healthy females (a control) for the content of some acute-phase proteins and cytokines to clarify the specific features of changes in their concentrations in relation to the type of the proliferative process. It was shown that in OC, there were significant reductions in the levels of alpha2-macroglobulin (MG), plasmin (PL), alpha1-antitrypsin (AT) and statistically significantly increases in the content of lactoferrin (LF), interleukin (IL)-6, IL-8, Ig, and the regulatory transport complex of P--M. In M, the concentrations of AT were lower and those of IL-6, IL-8, and PL-MG were higher (to a lesser degree than those in OC). In OCA, the levels of MG and IgA were increased; those of IL-8 and PL-MG were decreased. The concentrations of interferon and IgM were unchanged in all groups. The findings suggest that difefrent proliferative processes initially provoke a number of changes of varying magnitude and even directions in the serum levels of inflammation reactants, which should be borne in mind when conducting clinical tests in the intraoperative and, probably, postoperative periods.

[Acute inflammation phase reactants and interleukin-8 in myocardial infarction].

The study was undertaken to search for additional diagnostic criteria allowing the depth of myocardial damage to be estimated in males aged 57.2 +/- 9.6 years. Few interrelated acute phase reaction indices, including the levels of interleukin-8 (IL-8), lactoferrin (LF), alpha2-macroglobulin (alpha2-MG), plasmin (PL) and alpha2-MG-PL circulating complexes, were studied in serum on days 1, 7, and 17 of the onset of the disease. In small-focal myocardial infarction, the levels of alpha2-MG and PL were decreased on day 1 and those of LF and IL-8 were increased on day 14. On the contrary, in large-focal myocardial infarction, the concentrations of IL-8 and LF rose just on day 1 while those of alpha2-MG and PL remained unchanged. The detected differences may be used as additional criteria in differential diagnosis, particularly when ECG was of no informative value. Further, the concurrent elevation of alpha2-MG, PL, and PL-alpha2MG concentrations in large-focal myocardial infarction is indicative of poor prognosis.

[Cytokine and acute phase inflammation reactant levels in men with myocardial infarction].

The role of cytokines, their putative transporters, and other acute phase reactants was studied in 17 men aged 57.2 +/- 9.6 yr with microfocal myocardial infarction (MI) and in 21 ones with macrofocal MI. Control group comprised 30 age-matched men. Diagnosis of MI was confirmed by traditional functional tests and laboratory methods. It was shown that levels of TNF-alpha and MG-PL transport complexes were elevated regardless of MI type whereas acute phase reactants increased only in patients with macrofocal MI These patients also had enhanced IL-6 and IL-8 levels with gradual normalization of the former. In microfocal IM, IL-8 increased only by day 14. Inhibitory action of TNF-alpha appears to be insufficient to suppress effect of macrofocal IM but may be responsible for its selective effect on acute phase reactants. The above differences can be used as additional criteria for differential diagnosis of micro- and macrofocal MI especially in patients with repeated MI or His bundle block when traditional ECG data are of poor informative value.

[Some proteins of acute phase of inflammation in differential diagnostics of rheumatoid arthritis].

Levels of lactoferrin (LF), antithrombin-III (AT-III) and beta2-macroglobulin (MG) in blood of patients with rheumatoid arthritis (RA), reactive arthritis (ReA) and systemic lupus erythematosus (SLE) were examined for assessment their importance in differential diagnostics of the diseases. It was shown that on the average, LF and AT-III levels were increased at RA, while MG level was practically unchanged. At the same time LF level was stable high regardless of RA activity and duration degree, but its concentration was significantly increased also in ReA patients (33% patients). AT-III, on the contrary, depended on process activity and duration, was more specific to RA, than LF, but its sensitivity at RA was not enough high (from 25 to 44% patients with increased levels subject to disease duration and activity). Coefficient LFE/AT-III, obtained by multiplication of these proteins concentration, had the greatest diagnostic value. Its sensitivity at RA is on the average 85%, and specificity at differential diagnostics of RA--80% vs. ReA and 92% vs. SLE. We consider that coefficient LFbetaAT-III can be used as an additional criterion at differential diagnostics at RA early stages, while other specific antibodies cannot be detected yet.

[The serum levels of inflammatory reactants in women with inflammatory diseases of the uterine appendages, who participate in the in-vitro fertilization].

The blood concentrations of various inflammatory reactants were studied in female participants of the in-vitro fertilization (IVF) programs depending on the outcome. Comparison of the obtained values with the values characteristic of healthy women and with the levels of the above reactants in patients with subacute inflammatory diseases of the uterine appendages indicated that females with IVF failures had elevated levels of lactoferrin and circulating alpha2-macroglobulin-IgG complexes and decreased concentrations of IgG as in women with a localized pyodestructive process in the appendages, but in women having effective IVF, the levels of the study proteins did not differ from those in healthy donors. The authors recommend that the above indices should be used as criteria substantiating the need for additional sanitation and anti-inflammatory therapy before initiating the IVF program.

[The level of pregnancy-associated alpha2-glycoprotein and the hormonal background during different options of hormonal replacement therapy in the menopausal syndrome].

The authors studied the concentration of pregnancy-associated alpha2-glycoprotein (PA alpha2GP), a sensitive marker of estrogen-dependent tumors, and the association of its level with the serum content of a number of hormones: follicle-stimulating hormone, luteinizing hormone, estradiol (E-2), dehydroepiandrosterone sulfate (DEAS-S), and testosterone in females receiving the groups of drugs containing: 1) estradiol valerate; 2) 17beta-estradiol, and 3) tibolone. The type of the active ingredient of a drug and the duration of its administration were shown to differently affect both the concentration of hypothalamopituitary hormones and steroid sex hormones and the level of PA alpha2GP). The latter increased significantly in Group 1 and insignificantly in Group 2 and did not differ from the normal values in Group 3, at the same time the concentration of E-2 elevated in Groups 1 and 2, rather than in Group 3; the level of DEAS-S increased in Groups 2 and 3 irrespective of the duration of use. Moreover, there were elevated levels of testosterone in Group 3 and those of DEAS-S in Group 1 only when the drugs were administered for 3-6 months. A number of correlations were found in the levels of PA alpha2GP with those of steroid hormones. The authors consider that individual monitoring of the level of PA alpha2GP in the females who need hormonal therapy in menopause provides a useful guide to choosing a drug, monitoring its use efficiency, and preventing malignant proliferation in proper time.

[Alpha-2-macroglobulin ligands and their biotransport mechanisms].

Alpha-2-macroglobulin (MG) is a high-molecular weight glycoprotein that possesses a wide range of regulatory functions. Earlier it has been shown that covalent binding of MG with proteinases results in conformational transformation of MG, which enables MG to transport some additional types of cytokines linked by noncovalent interactions. The results of our study have demonstrated that the range of proteins, with the ability for additional binding with transformed MG is variable and comprises IgG, IgA, IgM, albumin, both types of lipoprotein chain, plasmin, some cytokines and even pregnancy associated alpha-2-glycoprotein (structured MG homolog). The major ligands are found to be albumin, IgG, plasmin and, to a lesser degree, lipoproteins. MG interactions with both acidic and low-alkaline proteinases contribute to neutralization of total charge of the formed complex at neutral pH, typical for internal fluids of the organism, and that the addition of low-density lipoprotein receptor-related protein (LRP) increases the amount of electroneutral complexes at pH 7.4. We suppose, that this mechanism enables the transformed MG (or may be its complex with other regulatory proteins) to rapidly precipitate rapidly on cellular surface and then, after binding with LRP and secondary neutralization of the total charge under physiological pH conditions, to pass through cellular membrane and to realize its own regulatory functions.

[The role of macroglobulin family proteins in the regulation of inflammation].

The macroglobulin family represents a group of universal regulators involved into control of the inflammatory response to external and internal pathogens. Alpha-2-macroglobulin (MG), the major protein in the family, has 3 different binding sites and high affinity to an endocytosis receptor that allows MG to participate in recognition and phagocytosis of the foreign agents. The macroglobulin family proteins are most powerful apoptosis inhibitors: they bind autoaggressive hydrolases accumulating during inflammation. MG is the main transporter of cytokines and growth factors controlling the inflammatory response. At the same time, the macroglobulins are negative reactants of an acute phase of inflammation and the decrease of their synthesis at later stages is necessary for stimulation of collagenosis processes, coagulation, activation of thymulin, stimulating NK. At septic inflammation binding of macroglobulins to exotoxins can localize inflammation, and initiate systemic inflammatory response. Macroglobulin damage sharply reduces their subsequent utilization; this provokes accumulation of makroglodulin-proteinase complexes in biological fluids and may cause immune inflammation.

[Specific reactions of polyfunctional proteins of the family of macroglobulins and lactoferrin with receptors and ligands in the protection of the body from pathogens].

The reactions of polyfunctional proteins alpha2-macroglobulin (MG) and lactoferrin (LF) lipoprotein-receptor protein (LRP) and different ligands (antibodies and hydrolases) for ascertaining the mechanisms of their interaction and the probable role of MG and LF in the neutralization of bacterial pathogens were under study. The binding of MG and LF with ligands, irrespective of their nature, was shown to lead to the neutralization of the complex thus formed. This mechanism of neutralization in reaction of LF with ligands was supposedly due to the antibacterial properties of this protein: multiple reactions of LF with receptor sites on the surface of pathogens neutralized the charge of the latter and facilitated phagocytosis. The interaction of LRP of immunocompetent cells with MG, in the presence of additional binding sites on its surface, made it possible for MG to effectively bind and present even heretofore unknown pathogens in the presence of hydrolases on their surface.