[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.

Interaction of HydSL hydrogenase from the purple sulfur bacterium Thiocapsa roseopersicina BBS with methyl viologen and positively charged polypeptides.

The effect of polypeptides having different charge on the activity of Thiocapsa roseopersicina HydSL hydrogenase was studied. Strong inhibition was shown for poly-L-lysine bearing positive charge. The inhibition was reversible and competitive to methyl viologen, an electron acceptor, in the reaction of hydrogen oxidation catalyzed by the hydrogenase. Peptides carrying less positive charge had weaker inhibiting effect, while neutral and negatively charged peptides did not inhibit the hydrogenase. Molecular docking of poly-L-lysine to T. roseopersicina hydrogenase showed strong affinity of this polypeptide to the acceptor-binding site of the enzyme. The calculated binding constant is close to the experimentally measured value (Ki = 2.1 µM).

[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.

[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.

[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.

The use of tenoten (pediatric formulation) in the therapy of chronic tension-type headaches in children.

Tension-type headaches are the most common type of headaches in children. No effective treatment for this disease is developed yet. After treatment with tenoten, the number of days with headache in 9-17-years-old children decreased by 40.9%, the mean headache intensity score decreased to 1.

[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.

[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.

[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.

[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.

[Hormones, cytokines and macroglobulines in blood of women with autoimmune thyroid diseases].

AIM: To study proportions of hormones, cytokines and macroglobulins in autoimmune thyroid diseases. MATERIAL AND METHODS: Participants of the study were 28 females aged 43 +/- 5 years with diffuse toxic goiter (DTG), 31 females aged 41 +/- 6 years with autoimmune thyroiditis (AIT) and 26 controls (n = 26) free of thyroid pathology aged 40 +/- 4 years. DTG and AIT diagnoses were confirmed by ultrasound and laboratory examinations. RESULTS: Both autoimmune diseases were accompanied by a significant rise of serum IL-6 and fall of TNF-alpha levels vs those in the controls. In AIT interferon-gamma was higher than in DTG while in DTG alpha-2-macroglobulin rose higher but TNF-alpha reduced more than in AIT. CONCLUSION: It is suggested that an elevated level of interferon-gamma in AIT contributes to blockade of endocytosis of peptid hormones and cytokines transported by macroglobulins. Overproduction of IL-6 in DTG may be caused by deficiency of its antagonist--TNF-gamma leading to an increase in macroglobulin concentration, its gene inductor is the above interleukin.

[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.

[Development of acute exudative inflammation in rats upon perftoran infusion].

The effect of the perftoran--a blood replacing emulsion with multifunctional properties--on the biochemical parameters of blood serum has been studied in rats with acute exudative inflammation. It is established, that perfloran does not influence the adaptive biochemical reactions on early terms of inflammation development. At the same time, perftoran essentially reduces exudate volume, however this effect is short-time upon single infusion of the preparation.

[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.

[Role of proteins of the macroglobulin family in regulation of tumor growth].

Proteins of the macroglobulin family are an ancient and evolutionarily conservative link of the immune system, which is actively involved in both inhibition of tumor growth cells and proliferation of tumor cells. Two basically different binding sites and a great conformational plasticity of all representatives of the macroglobulin family, as well as the presence of two to four representatives of the family in the blood of most species allow them to transport diverse substances and exert various regulatory influences on both the tumor and the entire organism. For example, the capacity of macroglobulins for binding hydrolases makes it possible to inhibit enzyme mediated tumor invasion. At the same time, an excess of macroglobulin/hydrolase complexes can activate apoptosis. The tumor is able of using macroglobulins, especially pregnancy-associated proteins, for its own protection. Specifically, pregnancy-associated alpha2-glycoprotein, which is actively produced by human tumor cells, blocks the histocompatibility complex antigens. On the contrary, the capacity of binding zinc stimulates the thymulin-dependent activation of natural killer cells. Nevertheless, the actively growing tumor expresses many receptors to macroglobulins, which are the main carriers of some cytokines and growth factors essential for proliferation.

[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.

The properties of hydrogenase from Thiocapsa roseopersicina.

The soluble and chromatophore-bound hydrogenases from the purple sulphur bacterium Thiocapsa roseopersicina strain BBS were purified up to homogeneity and the properties studied. The amino acid composition of hydrogenase preparations from different fractions of T. roseopersicina is identical, glycine and alanine as N-terminal amino acid residues. In comparison with other hydrogenases, especially in the immobilized state, the preparations obtained are shown to be more stable to O2 during storage and they are characterized by high thermal stability. Inactivation is observed above 78--80 degrees C and the optimal temperature for enzyme action is 70 degrees C. The homogeneous enzyme preparations catalyse the exchange reaction between 2H2 and H2O and reversible redox reactions of methyl viologen and benzyl viologen as well as H2 formation from reduced ferredoxin. According to our data, the hydrogenase of T. roseopersicina bound with chromatophores is identical to the soluble one.

Reversible hydrogenase of Anabaena variabilis ATCC 29413: catalytic properties and characterization of redox centres.

The catalytic and spectroscopic properties of the reversible hydrogenase from the cyanobacterium Anabaena variabilis have been examined. The hydrogenase required reductive activation in order to elicit hydrogen-oxidation activity. Carbon monoxide was a weak (Ki=35 microM), reversible and competitive inhibitor. A flavin with the chromatographic properties of FMN, and nickel were detected in the purified enzyme. A. variabilis hydrogenase exhibited electron paramagnetic resonance (EPR) spectra in its hydrogen-reduced state, indicative of [2Fe-2S] and [4Fe-4S] clusters. Although no EPR signals due to nickel were detected, the results are consistent with the enzyme being a flavin-containing hydrogenase of the nickel-iron type.

Redox properties and active center of phototrophic bacteria hydrogenases.

It is shown that the activity of phototrophic bacteria hydrogenases depends on the redox potential (Eh) of the medium. Hydrogenase from the purple sulfur bacterium Thiocapsa roseopersicina strain BBS reversibly activates H2 at Eh less than -290 mV (pH 7.0). When Eh is increased from -290 to -170 mV, the enzyme is converted into an inactive form which is accompanied by one-electron oxidation of its Fe-S cluster. In contrast, the hydrogenases of the purple nonsulfur bacterium Rhodobacter capsulatus B10 and the green sulfur bacterium Chlorobium limicola forma thiosulfatophilum exhibit maximum activity at Eh greater than -300 mV, favourable only for H2 uptake. When Eh decreases the activities of these enzymes drop dramatically; this accounts for their unidirectional effect directed mainly towards H2 uptake. Such dependence on Eh of activity of hydrogenases from these bacteria correlates with their physiological function in the metabolism of phototrophic bacteria, i.e. with the catalysis of the H2 uptake reaction. Hydrogenases from purple bacteria contain nickel and a single Fe-S cluster. Metal chelators do not affect the activity of these enzymes, which indicates that iron and nickel are tightly bound to the apoprotein. Sulfhydryl compounds irreversibly inactivate T. roseopersicina hydrogenase by 30-40% in the presence of sulfide. Acetylene and carbon monoxide are reversible inhibitors of the enzyme. EPR and inhibitory analysis indicate a direct interaction of H2 with the nickel ion in the active center of the T. roseopersicina hydrogenase.