Plasma ionization balance in chemical-picture and average-atom models.

We propose an approximate method to calculate ion partition functions in the context of the chemical-picture representation of plasmas as an interacting mixture of various ions and free electrons under the local-thermodynamic-equilibrium conditions. The method uses the superconfiguration approach and implies that the first-order corrections to the energies of excited electron configurations due to the electron-electron interaction may be replaced by a similar first-order correction to the energy of the basic configuration of an ion with the same number of bound electrons. The method enables one to significantly speed up the calculations and generally provides quite accurate results. Using the method proposed, plasma ionization balance and average ion charges calculated on the base of the chemical-picture representation show a good agreement with the relevant average-atom data. For the case of weak electron-ion nonideality, we provide approximate relations between the chemical-picture and average-atom values of the average ion charge, chemical potential, and plasma-density depression of ionization potential.

[Efficacy and safety of treatment with ampasse: the results of a randomized, double-blind, placebo-controlled trial in patients with chronic cerebrovascular disorders]. / Effektivnost' i bezopasnost' terapii preparatom ampasse: rezul'taty randomizirovannogo dvoinogo slepogo platsebo-kontroliruemogo issledovaniya u patsientov s khronicheskim narusheniem mozgovogo krovoobrashcheniya.

OBJECTIVE: To assess the efficacy and safety of the drug ampasse in the treatment of patients with chronic cerebrovascular disorders (CCVD). MATERIALS AND METHODS: A multicenter, randomized, double-blind, placebo-controlled, confirmatory study of the efficacy and safety of ampasse (phase III) was conducted in 124 patients aged 50 to 75 years. The main group (MG) - 62 patients, received the test drug ampasse, solution for intravenous administration, 5 mg/ml, at a dose of 5 ml (25 mg), intravenously bolus slowly, the duration of treatment was 15 days. Control group (CG) - 62 patients, received comparison drug: placebo (0.9% sodium chloride-5 ml). RESULTS: All 124 patients fully completed the procedures and visits, there were no dropouts from the study. The proportion of patients who reached the primary endpoint (an increase in the score by 2 or more points on the MoCA scale) was 83.87% in MG and 22.58% in CG, that is, the efficacy of therapy in MG was 61.29% higher than in CG (p<0.001), and good tolerability of the drug was proved. The secondary endpoint is an increase in quality of life (QOL) on the SF-36 V2 scale on Day 31. In MG, there was a statistically significant improvement in all indicators of QOL compared to the baseline. When assessing the safety spectrum, the proportion of patients who had adverse events was 14.52% in MG and 8.06% in CG (p=0.395). CONCLUSION: Ampasse has a positive effect on cognitive functions and QOL, does not increase the frequency of adverse events in patients with CCVD compared to placebo, does not cause significant side effects, and is well tolerated by patients.

The role of SAGA coactivator complex in snRNA transcription.

The general snRNA gene transcription apparatus has been extensively studied. However, the role of coactivators in this process is far from being clearly understood. Here, we have demonstrated that the Drosophila SAGA complex interacts with the PBP complex, the key component of the snRNA gene transcription apparatus, and is present at the promoter regions of the snRNA genes transcribed by both the RNA polymerase II and RNA polymerase III (U6 snRNA). We show that SAGA interacts with the Brf1 transcription factor, which is a part of the RNA polymerase III transcription apparatus and is present at promoters of a number of Pol III-transcribed genes. Mutations inactivating several SAGA subunit genes resulted in reduced snRNA levels in adult flies, indicating that SAGA is indeed the transcriptional coactivator for the snRNA genes. The transcription of the Pol II and Pol III-transcribed U genes was reduced by mutations in all tested SAGA complex subunits. Therefore, the transcription of the Pol II and Pol III-transcribed U genes was reduced by the mutations in the deubiquitinase module, as well as in the acetyltransferase module of the SAGA, indicating that the whole complex is essential for their transcription. Therefore, the SAGA complex activates snRNA genes suggesting its wide involvement in the regulation of gene transcription, and consequently, in the maintenance of cellular homeostasis.

[Interactions of the TREX-2 complex with mRNP particle of ß-tubulin 56D gene].

mRNA transport from the nucleus to the cytoplasm is an essential step of eukaryotic gene expression. A pre-mRNA molecule undergoes modification, such as 5'-capping, splicing, and 3'-end processing, in the nucleus. The molecule being modified interacts with a large number of proteins and, thus, mRNP particles are formed. The binding of factors involved in nuclear export also occurs during transcription and mRNA processing. We have shown that the functioning of TREX-2, an mRNA export complex, is restricted to the nucleus. We used the method of RNA coprecipitation that enables the selective extraction of RNA-protein complexes from samples to show that the transcription elongation complex TREX interacts with mRNA of the ß-tubulin 56D gene over the entire length of the molecule. The capping protein Cbp80 reacted both with the cap structure and with a specific part of the coding mRNA of the ß-tubulin 56D gene. The TREX-2 complex that mediates mRNA export from the nucleus to the cytoplasm is bound to the same part of the coding sequence. Thus, we identified a common binding site for all of the complexes under investigation on the mRNA of ß-tubulin 56D. Co-immunoprecipitation reactions performed with S2 cell extracts revealed interactions between the components of complexes involved in transcription elongation, maturation, and export of mRNA. The model of molecular folding for the mRNP particle involving the mRNA of ß-tubulin 56D has been proposed.

Some properties and possible biological role of peptidase inhibitors from the entomopathogenic fungus Tolypocladium cylindrosporum.

The activities of secreted and mycelial inhibitors of proteolytic enzymes from fungi of the order Hypocreales have been investigated. Inhibitors of bromelain, papain, and trypsin of low molecular mass (about 1 kDa) and a subtilisin proteinaceous inhibitor with molecular mass of 45 kDa were revealed in the culture liquid of the fungus Tolypocladium cylindrosporum. The subtilisin inhibitor from T. cylindrosporum has antibiotic properties, significantly decreased the activity of purified bacterial enzymes, and prevented the growth of the bacterium Pseudomonas sp. Data suggesting the existence in fungi of the Hypocreales order of two pools of peptidase inhibitors have been obtained.

[Methods to study the RNA-protein interactions].

RNA-binding proteins (RBPs) play an important role in regulating gene expression at the posttranscriptional level, including the steps of pre-mRNA splicing, polyadenylation, mRNA stabilization, mRNA export from the nucleus to the cytoplasm, mRNA localization, and translation. RBPs regulate these processes primarily by binding to specific sequence elements in newly synthesized or mature transcripts. While many RPBs are known to recognize certain nucleotide sequences in RNA, information is insufficient for others. In particular, RBPs often compete for RNA binding or interact with RNA cooperatively. Hence, it is of importance to study the RNA-protein interactions in vivo. Numerous methods have been developed to identify the target nucleotide sequences of RBPs. The methods include the electrophoretic mobility shift assay (EMSA), systematic evolution of ligands by exponential enrichment (SELEX), RNA pull-down assay, RNA footprinting, RNA immunoprecipitation (RIP), UV-induced crosslinking immunoprecipitation (CLIP) and its variants, and measurement of the level for newly synthesized transcripts. Each of the methods has its limitation, and several methods supplementing each other should be employed in order to detect the RNA sequence to which a protein binds.

Fungal inhibitors of proteolytic enzymes: classification, properties, possible biological roles, and perspectives for practical use.

Peptidase inhibitors are ubiquitous regulatory proteins controlling catalytic activity of proteolytic enzymes. Interest in these proteins increased substantially after it became clear that they can be used for therapy of various important diseases including cancer, malaria, and autoimmune and neurodegenerative diseases. In this review we summarize available data on peptidase inhibitors from fungi, emphasizing their properties, biological role, and possible practical applications of these proteins in the future. A number of fungal peptidase inhibitors with unique structure and specificity of action have no sequence homology with other classes of peptidase inhibitors, thus representing new and specific candidates for therapeutic use. The main classifications of inhibitors in current use are considered. Available data on structure, mechanisms and conditions of action, and diversity of functions of peptidase inhibitors of fungi are analyzed. It is mentioned that on one side the unique properties of some inhibitors can be used for selective inhibition of peptidases responsible for initiation and development of pathogenic processes. On the other side, general inhibitory activity of other inhibitors towards peptidases of various catalytic classes might be able to provide efficient defense of transgenic plants against insect pests by overcoming compensatory synthesis of new peptidases by these pests in response to introduction of a fungal inhibitor. Together, the data analyzed in this review reveal that fungal inhibitors extend the spectrum of known peptidase inhibitors potentially suitable for use in medicine and agriculture.

[Hormones of adipose tissue in diabetes mellitus and its complications (review of literature and the authors'own data)].

The contemporary ideas on endocrine function of the adipose tissue and its role in pathogenesis of diabetes mellitus and insulin resistance associated with it and the metabolic syndrome are shown in the review. A change in the life style (excessive and irrational nutrition, insufficient physical loading, psychological disorders) and also the reduction of genetic and immunologic controlling mechanisms contribute to the development of obesity, that is now considered as low-grade inflammation. An increased number of small size adipocytes and macrophages of the adipose tissue begin to secrete an increase number of proinflammatory adipocytokines and chemokines that result in the inflammatory and metabolic stress accompanied by the stimulation of signal pathways, leading to increased insulin requirement, on the one hand, and promoting to the beta-cell death, on the other hand. The role of some adipocytokines such as IL-6, TNF-alpha, leptin, adiponectin, visfatin and resistin was demonstrated in these processes.

[The level of circulating cytokines and chemokines in the preclinical and early clinical stages of type IA diabetes mellitus development].

AIM: to study the level of circulating proinflammatory (IL-1alpha, IL-1beta, IL-6, alpha-TNF, IFN-gamma) and anti-inflammatory (IL-4, IL-10) cytokines and chemokines (IL-8, IL-16) in preclinical development of type 1A diabetes mellitus (T1DM) in children. SUBJECTS AND METHODS: An examination was made in 450 children who had normal blood glucose levels and a burdened history of positive or negative Langerhans islet autoantibodies (LIAA): IAA, GADA, and 1A-2A over time until the clinical manifestations of DM1 emerged. The levels of the cytokines and chemokines were determined by ELISA and the titer of LIAA was by radioimmunoassay. RESULTS: Long before T1DM was clinically diagnosed, most children with normal blood glucose levels and LIAA had elevated levels of the cytokines IL-1alpha, IL-6, and alpha-TNF and the chemoattractants IL-8 and IL-16 with lower IL-4 concentrations as compared with the similar indices in children without LIAA and controls. After the disease manifested, the magnitude of changes in the indices under study reduced in the majority of children with LIAA, which may suggest that the autoimmune process subsides after destruction of most beta-cells. CONCLUSION: The elevated levels of IL-6, IL-16, alpha-TNF, and the chemokine IL-8 with the lower blood content of the cytokine IL-4 were long before the development of DM1 in children with normal blood glucose level in the presence of LIAA, which should be borne in mind while developing the immune mechanisms specifically directed against block, which participate by means of cytokines in beta-cell destruction, as well as methods for preventing the development of T1DM in subjects with LIAA.

[Circulating interleukin-16 in blood of patients with metabolic syndrome and type 2 diabetes mellitus].

The authors determined interleukin-16 (IL-16) content in blood serum of patients distributed into three groups using an immunoenzymic method (ELISA). The first group consisted of patients with type 2 DM and metabolic syndrome (MS); the second group with type 2 DM and without MS, the third group - with MS and without T2DM. The control group consisted of normoglycemic subjects without MS signs who were distributed into two subgroups: 1) with excessive weight; 2) with normal weight. A significant increase in IL-16 concentration in blood serum was noted in patients with T2DM associated with MS (249,5+/-75,3 pg/ml) versus patients with MS without T2DM (130,5+/-41,2 pg/ml), and versus patients with T2DM without MS (69,5+/-35,6 pg/ml, P<0,05) and without obesity (77,4+/-11,6 pg/ml, P<0,05). This increase correlated with abdomen volume (r=0,4, P<0,05) and triglyceride level (r=0,4, P<0,05).

[Level of circulating interleukin-16 during preclinical stage of type I diabetes mellitus development in children].

The content of IL16 in blood serum of children with type I diabetes mellitus and healthy normoglycemic children with genetic burden of diabetes mellitus and positive to autoantibodies against the islets of Langerhans (GADA and IAA) was determined by the immunoenzyme method ELISA. Healthy normoglycemic children with genetic burden but negative to GADA and IAA and healthy normoglycemic children without genetic burden but negative to GADA and IAA have been observed. The conducted examinations established that a statistically significant increase in the content of IL-16 and its decrease in developed diabetes mellitus patients have been noted to have place among children with genetic burden to diabetes, positive to GADA and IAA in comparison with control groups.

[The immunity during the pre-clinical period of type I diabetes mellitus].

The purpose of the study was to investigate the condition of immunity (blood lymphocyte immune phenotype and ultrastructure) in healthy children with a family background of type 1 diabetes mellitus (DM 1) having or not having diabetes-associated autoantibodies (DAAB). The subjects of the study were divided into three groups. Group 1 consisted of 90 children with a family background of DM 1 (first line relatives had DM 1), DAAB- (GADA, IA-2A, and IAA) positive or negative; group 2 consisted of 51 children with newly revealed DM 1; group 3 included 45 healthy controls, normoglycemic DAAB-negative children with no family background of DM 1. GADA, IA-2A, and IAA titers were measured using radioimmunoassay. The immune phenotype of lymphocytes (CD3+, CD4+, CDr8, CD20+, and CD56+ cells) were studied using flow cytometry (FACS-analysis); their ultrastructure was studied by means of electron microscopy. The study found a significantly lower total number of T-lymphocytes (CD3+ cells), T-helpers/inductors (CD4+ cells), and natural killer cells (CD56+ cells and large granule-containing lymphocytes) in the DAAB-positive children vs. the DAAB-negative ones and especially the controls. In the DAAB-positive children, electron microscopy found distinct changes in the ultrastructure of CD4+ lymphocytes and large granule-containing lymphocytes (CD56+ cells), which evidences changes in the secretory and cytostatic function. Such changes in the number and ultrastructure of these lymphocyte subpopulations are found in patients with newly revealed DM 1. Thus, immune changes happen in the organism of a healthy person a long time before clinical manifestations of DM 1 develop; these changes reflect a concealed autoimmune process in Langerhans islets. Detection of DAAB plays a significant role not only in studying poorly understood pre-diabetes nature, but also in the development of new, scientifically based methods of its prevention and treatment.

[Discovery of autoantibodies to Langergan's islands of the pancreas is a prominent achievement in the field of forecasting the development and diagnostics of diabetes mellitus in clinics].

The authors have summarized literature data and results of own studies on autoantibodies and beta-cells of Langergan's islands of the pancreas (ICA, GADA, IA-2A and IAA). It may help predicting the development of I type diabetes mellitus even in practically healthy individuals before they present clinically evident disease. The history of the discovery of these autoantibodies, their immunological properties, comparative characteristics, sensibility and specificity and their application in clinical practice to forecast the development of I type diabetes mellitus in different populations of children and adolescents as well as to more exactly carry out differential diagnostics of DM type I and DM type II in difficult cases in adults is presented in the article. Having diabetes-associated autoantibodies as a diagnostic tool allows to diagnose a separate subtype of diabetes mellitus--autoimmune diabetes mellitus of adults (LADA) and it is very important to choose a right way of the treatment.

[The ultrastructure and function of lymphocytes in children with first detected untreated type 1 diabetes mellitus].

Flow cytometry (FACS-analysis), light and electron microscopy, and cytotoxic test were used to study the leukocytic composition, content, ultrastructure, and function of different lymphocytic populations (CD3+, CD4+, CD8+, CD20+, and CD56+ cells) in the blood of 90 children aged 8 to 15 years who had untreated new-onset type 1 diabetes mellitus. A control group consisted of 45 apparently healthy normoglycemic children. The sick children were observed to have a slight, but statistically significant reduction in the relative and absolute blood content of CD3+, CD4+, and CD56+ cells, CD4/CD8 index and particularly large granule-containing lymphocytes (the morpho-logical homologue of natural Idller (NK) cells). Ultrastructurally, CD20+4 cells (lymphocytes containing Gall's bodies) showed the signs of a high functional activity while NK cells (large granule-containing lymphocytes) exhibited a low one, which was confirmed by the decreased cytotoxic activity of these cells determined in vitro. Three-month insulin therapy leading to the restoration of the blood content of glucose and glycosylated hemoglobin fails to normalize the detected changes in the parameters of T- and NK-cell immunity.

[Bicarbonate calcium mineral water with carbon dioxide in rehabilitation of children with dismetabolic nephropathies complicated by renal inflammation]. / Uglekislaia gidrokarbonatnaia kal'tsievaia mineral'naia voda v reabilitatsii detei s dismetabolicheskimi nefropatiiami v sochetanii s vospalitel'nymi zabolevaniiami pochek.

A renal function was studied in children with dismetabolic nephropathy and renal inflammation before and after spa treatment with low-mineral water from the spring "Gornovodnoye". Drinking the water resulted in intensification of 24-h diuresis in increasing proportion of sodium and chlorine ions concentrations and decreasing proportion of calcium and magnesium ions concentrations. A membrane-stabilizing action of Gornovodnenskaya mineral water reduced oxaluria and uraturia. An effective scheme of balneotherapy is proposed.

[The use of intraaortic balloon counterpulsation in patients with acute myocardial infarction]. / Primenenie vnutriaortal'noi ballonnoi kontrpul'satsii u bol'nykh ostrym infarktom miokarda.

Intraaortic balloon counterpulsation was implemented in 11 patients with acute myocardial infarction and 1 patient with unstable angina. All patients had severe multivessel coronary artery disease. In 9 patients counterpulsation was used in conjunction with percutaneous transluminal coronary angioplasty, in 3 - with thrombolytic therapy. During hospitalization 2 patients died of progressing heart failure, while significant improvement of hemodynamic parameters occurred in other patients. Thus intraaortic balloon counterpulsation used in combination with angioplasty and thrombolytic therapy is an easily accessible highly effective method of treatment of cardiogenic shock in patients with acute myocardial infarction

[Considerable decrease in chemokine interleukin-16 level in blood of children with diabetes mellitus type I diagnosed for the first time]. / O znachitel'nom snizhenii urovnia khemokina interleikina-16 v krovi detei s vpervye vyiavlennym sakharnym diabetom tipa 1.

Considerable decrease in chemokine interleukin-16 level has been detected in blood serum of children with the first diagnosed in life diabetes mellitus type I.

[Content of different cytokines in the blood of healthy children and their siblings who are either.positive or negative for diabetes-associated autoantibodies (GADA, 1A-2A, IAA)]. / Soderzhanie razlichnykh tsitokinov v krovi zdorvykh detei-sibsov, pozitivnykh i negativnykh po nalichiiu diabetassotsirovannykh autoantitel (GADA, 1A-2A, IAA).

Content of different cytokines (IF alpha, TNF alpha, IL-1 alpha, IL-4, IL-6 and IL-10) was examined in the blood serum in two groups of healthy children-siblings with type 1 diabetes mellitus with and without revealed insulin autoantibodies against pancreatic islets (GADA, IA-2A and IAA) by enzyme-like immunosorbent assay (ELISA). In the group of patients with two and more revealed autoantibodies the higher indices in the number of IF alpha, TNF alpha and IL-6, and the decrease in the level of IL-4 comparing with the group of children with negative reaction to diabetes associated autoantibodies were more often observed.

[Lethal outcome in a patient with pulmonary hypertension and liver cirrhosis on the 12-th week of pregnancy]. / Letal'nyi iskhod u bol'noi s legochnoi gipertenziei i ochagovym tsirrozom pecheni v 12 ned beremennosti.

A patient with pulmonary hypertension and focal liver cirrhosis was hospitalized on the 12-th week of pregnancy because of threatened abortion and died after 2 days of hospital stay during which symptoms of portal and pulmonary hypertension progressed, decompensation of pulmonary heart and hepatic cellular insufficiency ensued and syndrome of disseminated intravascular coagulation developed.

[Blood plasma level of insulin and sensitivity to it in tissues of patients with newly detected diabetes mellitus type II]. / Soderzhanie insulina v plazme krovi i chuvstvitel'nost' k nemu tkanei u bol'nykh s vpervye vyiavlennym sakharnym diabetom II tipa.

Type II diabetes mellitus is the most common type of the condition. But little is yet known about mechanisms responsible for its development in persons with different body masses. We studied blood plasma levels of insulin and assessed sensitivity to it in 32 patients with freshly detected type II diabetes mellitus with normal and above normal body weight. Of these, 20 subjects were obese, the other 12 being not fat; the control group comprised 30 persons obese and non-obese, with n = 15 and 15 respectively. Recordable in patients was a marked elevation of blood plasma level of insulin accompanied by decrement of the peripheral tissue sensitivity to it in obese diabetic patients while in those persons with normal body mass there was a substantial reduction in the production of insulin, which differences may indicate different pathogenesis of development of type II diabetes mellitus in persons with different body masses.