[Therapy of moderate cognitive impairment and asthenia in patients with cerebrovascular pathology: results of a prospective observational study]. / Terapiya umerennykh kognitivnykh narushenii i astenii u patsientov s tserebrovaskulyarnoi patologiei: rezul'taty mnogotsentrovoi otkrytoi prospektivnoi nablyudatel'noi programmy.

OBJECTIVE: Obtaining additional data on the efficacy and safety of the drug Prospekta in the treatment of moderate cognitive impairment (MCI) and asthenia in patients with cerebrovascular disease (CVD). MATERIAL AND METHODS: A prospective observational study in more than 40 Russian cities enrolled 232 patients (mean age 61.5±10.0 years) with mild cognitive impairment (MCI), asthenia on ongoing basic nootropic therapy. The presence of MCI was confirmed by the Montreal Cognitive Assessment Scale (MoCA), asthenia - by 10-point Visual Analog Scale (VAS). All patients were prescribed the nootropic medication Prospekta 2 tablets 2 times a day for 8 weeks in addition to the therapy they received. Ultrasound Doppler sonography of the main arteries of the head and magnetic resonance imaging (MRI) of the brain were also assessed. At the end of treatment, the Clinical Global Impression Efficacy Index (CGI-EI) was assessed and the safety of the treatment was evaluated. RESULTS: The baseline severity of cognitive impairment according to the MoCA scale was 21.6 points, severity of asthenia according to the VAS was 6.3 points. According to Doppler flowmetry findings, hemodynamically significant stenosis was revealed in 105 (49.3%) patients, and narrowing of the main vessels without changes in hemodynamic parameters was revealed in 108 (50.7%) patients. According to MRI results, single vascular lesions in the brain matter were detected in 102 (44.0%) patients. The medications with nootropic effect were administered to 144 (62.1%) patients. A positive therapeutic response as improvement of cognitive functions was seen in 93.3% of patients after 8 weeks of taking Prospekta, including 39.4% of patients who had cognitive functions restored to the normal level. No side effects were registered during the observational study. CONCLUSIONS: The nootropic medication Prospekta is effective and safe in treatment of MCI in patients with asthenia with CVD, and improves cognitive function in patients with asthenia with CVD, both in monotherapy and in combination with other nootropic agents.

[Biological age and the pain syndrome at diabetic polyneuropathy].

Patients with diabetic polyneuropathy were examined to study their biological age, rate of aging and pain syndrome. More rapid rate of aging was revealed in patients with diabetic polyneuropathy and pain syndrome. Using Duloxetin and Gabapentin is reliable to decrease the display of pain syndrome in diabetic polyneuropathy patients. Against the background of pain syndrome therapy the rate of aging is noticed to decrease along with the regression of pain syndrome. The medicines of Duloxetin and Gabapentin which are used during depression and epilepsy protect against aging.

A cyclic adenosine 3',5'-monophosphate-dependent histone kinase from pig brain. Purification and some properties of the enzyme.

A cyclic adenosine 3',5'-monophosphate-dependent histone kinase (ATP: protein phosphotransferase, EC 2.7.1.37) was isolated from pig brain. The enzyme has been purified 1140-fold; it is homogeneous on polyacrylamide gel electrophoresis and gel filtration. The estimated molecular weight of the enzyme is 120 000. Histone kinase dissociates into a catalytic subunit and a regulatory one (molecular weights 40 000 and 90 000, respectively). The catalytic subunit has been obtained in homogeneous state as evidenced by sodium dodecylsulphate-polyacrylamide gel electrophoresis. At all purification steps, enzymatic activity is stimulated 5-fold by cyclic AMP. An apparent Km value for cyclic AMP is about 3.3 - 10- minus 7 M. In the presence of cyclic AMP(5 - 10- minus 6 M), the Km value for ATP and F1 histone were 1.2 - 10- minus five and 3 - 10- minus 5 M, respectively. Optimum pH value for histone kinase is 6.5, its isoelectric point is situated at pH 4.6. The purified enzyme displays high specificity for the lysine-rich and moderately lysine-rich histones F1, F2a2 and F2b. Arginine-rich histones and other known protein substrates for cyclic AMP-dependent protein kinases (casein, Escherichia coli RNA polymerase, etc.) are extremely poor substrates for this enzyme.

Investigation of the adenosine 3', 5'-cyclic phosphate binding site of pig brain histone kinase with the aid of some analogues of adenosine 3', 5'-cyclic phosphate.

2'-O-Chloroacetyl cyclic AMP, 2'-O-acrylyl cyclic AMP and N-6, 2'-O-diacrylyl cyclic AMP were synthesized by the reaction of cyclic AMP with chloroacetic and acrylic anhydrides, respectively. Selective O-deacylation of N-6, 2'-O-diacrylyl cyclic AMP yielded N-6 -monoacrylyl cyclic AMP. In the reaction of gamma-mercaptobutyric acid with 8-bromo cyclic AMP, 8-(gamma-carboxypropylthio) cyclic AMP was obtained. The compounds synthesized and other cyclic AMP analogues (8-bromo cyclic AMP and adenosine 3', 5'-cyclic sulphate) were tested for ability to interact with the highly purified pig brain histone kinase. All compounds under study were found to be activators of the enzyme. The highest activating potency was manifested by 8-bromo cyclic AMP and 8-(gamma-carboxypropylthio) cyclic AMP; adenosine 3', 5'-cyclic sulphate was the least potent in this respect. All compounds were shown to inhibit binding of cyclic [-3-H]AMP to histone kinase. The inhibition was competitive with respect to cyclic AMP in all cases. All compounds, except for 2'-O-chloroacetyl cyclic AMP may indicate the formation of a covalent bond between this analogue and the enzyme. These findings suggest that an active site of the regulatory subunit of the histone kinase contains at least three specific areas responsible for cyclic AMP binding.

The autophosphorylation reaction in the mechanism of activation of pig brain cyclic AMP-dependent protein kinase.

Autophosphorylation of cyclic AMP-dependent protein kinase (ATP: protein phosphotransferase, EC 2.7.1.37) was shown to occur via an intramolecular mechanism: the regulatory subunit undergoes phosphorylation only within the holoenzyme. The phospho form of the catalytic subunit has the capacity to phosphorylate the regulatory subunit. The phosphotransferase reaction and the reaction of autophosphorylation were found to proceed with the involvement of the same active site. The activation constant of phospho- and dephosphoprotein kinase under the influence of cyclic AMP and the dissociation constant of the cyclic AMP complex with phospho- and dephospho forms of the holoenzyme were estimated. Autophosphorylation was demonstrated to lead to almost complete dissociation of the holoenzyme under the influence of cyclic AMP. Circular dichroism spectra of the phosphorylated and non-phosphorylated forms of protein kinase were studied. The relative content of the secondary structure elements in proteins was estimated and conformational changes were detected in the enzyme upon its interaction with cycli AMP. The anti-conformation of the cyclic nucleotide fixed in the complex with the phospho form of the regulatory subunit is suggested.

Protein kinase A: regulation and receptor-mediated delivery of antisense oligonucleotides and cytotoxic drugs.

Protein kinases help regulate eukaryotic cell division. We investigated the regulation of cAMP-dependent protein kinase A (PKA) and casein kinase (CK) type I activity in normal cells and in cancer. To assess this activity in biopsies we suggest a new parameter--the ratio of CK activity and total PKA activity divided by cAMP concentration: CK/PKA/cAMP. In 98 samples of colon mucosa in normal, inflamed, polyp, and adenocarcinoma cells, we found this parameter to be fairly constant in normal conditions and increased 10-fold in colon cancer; the ratio does not depend on the place of biopsy or the patient's age or sex. Experiments with model systems of concanavalin A-stimulated lymphocytes and regenerating rat liver showed that in normal cell proliferation the parameter increases 2-3-fold, as compared to a 30-fold increase in cancer. Unlike normal cells, malignant cells show CK activation and decrease of cAMP; therefore, PKA activity decreases. This suggests a correlation of CK and PKA activity and significant damage to their regulation at pathological changes of tissue proliferation. To further study concerted CK and PKA regulation we used monoclonal antibodies (mAbs) against cAMP-dependent protein kinase regulatory subunit RKII beta. We produced 11 antibodies in three groups: inhibiting, which block cAMP binding with RII beta and inhibit holoenzyme formation (RS6); activating, which enhance cAMP binding and do not affect holoenzyme formation (RS28); and neutral (RS17). To investigate mAb influence on protein kinase regulation in live cells we permeabilized pheochromocytoma PC12 by digitonin. When used at 5-microM concentration for 5 min, digitonin allowed us to deliver mAb into PC12 cells at 30-34-nM concentration, leaving 68-75% viable cells. Protein kinase activity was measured within 0.5 and 4 h after incorporation of mAbs into cells. After 30 min incorporation, mAb RS6 blocked PKA activation in PC12 cells under the influence of cAMP; other mAbs showed no effect. mAb RS6 caused a 4-fold increase of free C subunit activity 4 h after incorporation. mAb RS38 decreased R2C2 activity and did not influence C subunit activity. The change of free C subunit activity caused by mAb incorporation was followed by a synchronized, well-balanced change of CK type I activity, which suggests a correlation between the two phosphorylation systems of cell proteins.

Nanoscale iron(III) oxyhydroxy aggregates formed in the presence of functional water-soluble polymers: models for iron(III) biomineralisation processes.

Superparamagnetic clusters of iron(III) oxyhydroxide in the form of poorly crystalline ferrihydrite (formally, 5Fe2O3 x 9H2O) have been synthesised in the presence of the polymers polyvinyl alcohol (PVA), polyacrylic acid (PAA) and alginic acid. The solutions have been characterised by viscosity studies and the resultant arrays isolated from these solutions have been imaged under an electron microscope and their magnetic properties determined by 57Fe-Mössbauer spectroscopic studies at 293 and 77 K and magnetisation measurements at 293 and 5 K. The magnetic data show that the iron(III) oxyhydroxy particles are superparamagnetic. All preparations show hysteretic behaviour with coercive fields being approximately half or less than half of that of ferrihydrite (3.4 kOe) and values of magnetic moment per iron particle less than that of ferrihydrite. Nanoscale aggregates (2-4 nm) are formed in the presence of PVA and PAA while, with alginic acid, extended branch-like structures are observed, their formation being facilitated by the comparatively rigid polysaccharide chain, a process related to iron biomineralisation in diverse biological systems.

[Influence of histone kinase phosphorylating lysine-rich histones, on the physico-chemical properties of normal hepatocyte chromatin and after partial hepatectomy]. / Vliianie gistonkinazy, fosforiliruiushchei bogatye lizinom fraktsii gistonov, na fiziko-khimicheskie svoistva khromatina gepatotsitov v norme i posle chastichnoi gepatéktomii.

The influence of a specific histone kinase, phosphorylating lysin-rich histone H1, H2a, H2b on the physico-chemical properties of chromatin from hepatocytes of normal and hepatectomized guinea pigs has been investigated. A cytochemical method has been used which permits to obtain information about the physico-chemical properties of the chromatin in situ, i.e. without its isolation. This approach allows us to evaluate changes in chromatin properties in cell cultures as well as in the intact organism. It is found that the specific histone kinase changes the properties of chromatin in non-dividing cells bringing about an increase of acridine orange binding to the level characteristic for hepatocytes after partial hepatectomy. At the same time the chromatin properties in activated hepatocytes are not changed under the action of the histone kinase. It is concluded that the specific histone kinase, phosphorylating lysine-rich histones can play an important role in the course of chromatin activation in cells stimulated to proliferation.

[Phosphorylation of lysine-rich histones by swine brain histokinase]. / Fosforilirovanie bogatykh lizinom gistonov gistokinazoi iz mozga svin'i

The phosphorylation of lysine-rich histones F1, F2a2 and F2b of calf thymus has been investigated using homogeneous histone kinase from pig brain. 32P-labelled phosphopeptides from tryptic digests of corresponding histones were obtained. According to N-terminal analysis and the quantitative determination of amino acid composition of the obtained radioactive peptides the sites of phosphorylation were identified in the primary structure of lysine-rich histones, namely, Ser-38 for the polypeptide chain of histone F1, Ser-19 or 18 for histone F2a2 and Ser-14 and 36 for histone F2b. Thus, the high specificity of brain histone kinase in vitro was demonstrated.

[Immuno-microbiological aspects of chronic bacterial prostatitis pathogenesis]. / Immunomikrobiologicheskie aspekty patogeneza khronicheskogo bakterial'nogo prostatita.

The paper deals with a role of the factors of the body's antiinfective resistance system in the pathogenesis of chronic bacterial prostatitis and with the identification of an etiological factor of the disease. Long-term studies have revealed that chronic bacterial prostatitis runs in the presence of impairments in the antiinfective resistance system, in failures of T lymphocytes, phagocytic activity, and opsonization in particular. Gram-positive cocci, predominantly Staphylococcus and Enterococcus alone or as part of 2-component associations have been found to be the leading agents in the etiology of chronic bacterial prostatitis.

[Human microflora in the norm and in pathology studied with laser fluorescence]. / Indikatsiia mikroflory cheloveka v norme i patologii s pomoshch'iu lazernoi fluorestsentsii.

On the basis of experimental and clinical data the use of laser fluorescence for indication of microflora in normal and pathological states has been substantiated. Standardized intensity characteristics of fluorescence integrally reflect the presence and activity of the total microflora and its changes in dysbiosis and pyo-inflammatory diseases. The possibility of its use in clinical practice is shown.

[Monitoring of etiological agents of acute enteric infections in the Moscow region]. / Monitoring vozbuditelei ostrykh kishechnykh infektsii v regione Moskovskoi oblasti.

The fecal microflora of patients with acute enteric infections (AEI) has been examined on the territory of the Moscow region. The pathogens of high, moderate and low priority levels have been detected. As revealed in this study, shigellae, salmonellae, enteropathogenic and enteroinvasive Escherichia are the etiological agents of bacterial diarrheas on the territory of the Moscow region. The dynamics of the etiological agents of AEI in the region has been established.

[Cyclic nucleotides in the regulation system of cell proliferation and differentiation]. / Tsiklicheskie nukleotidy v sisteme reguliatsii kletochnoi proliferatsii i differentsirovki.

The paper deals with certain aspects in the role of cyclic nucleotides in cell differentiation and proliferation. The content of cyclic nucleotides and activity of protein kinases are determined for the cell cycle stages of a synchronous plasmodium culture of true myxomycete Physarum polycephalum. Studies in the biological role of the brain cAMP-dependent protein kinase show that during phosphorylation of nuclei from the brain and subsequent isolation of chromatin its matrix activity is higher than the control one. When a regulatory subunit is added to the standard RNA-polymerase reaction, the total level of transcription increases for eucaryote RNA-polymerase. The constant of the regulatory subunit binding with chromatin from the brain is 10(10) M-1. Studies in translocation of tritium-labelled cAMP-dependent protein kinase and its subunits to interphase nuclei and in acception of these proteins by metaphase chromosomes in different cell cultures state that sAMP-dependent protein kinase penetrates into the nucleus in the dissociated state. It is established that disturbance of the regulatory subunits translocation into the nucleus and its acception by chromosomes when cell are transformed by SV40 virus is completely restored by adding cAMP. For cells, which suffered the malignant transformation of another type (spontaneous human cancer--KB, HeLa), and essential nuclear translocation and a complete absence of acception by chromosomes for the regulatory subunit are found. An assumption is made that compartmentation of cAMP-dependent protein kinase, in particular of its regulatory subunit, plays an important role in the organism vital activity and disturbances arising in neoplastic transformation are connected with changes in translocation and acception of the regulatory subunit in the nucleus.

[Monitoring of uropathogens and their susceptibility to antibiotics]. / Monitoring uropatogenov i ikh chuvstvitel'nosti k antibiotikam.

Samples of urine collected from patients with complicated urology infection and hospitalized to the Moscow Region Research Clinical Institute in 1986, 1991, 1995 and 1999 were analysed. Of 11,444 samples examined, bacteriuria was estimated in 7143 samples. 9786 strains (29 genus) of bacteria were isolated--56.9 per cent as mono culture and 43.1 per cent as associations. Susceptibility to 21 antibiotic was determined by disk diffusion method for 1607 strains; beta-lactamase production was determined in 198 strains, MIC was determined for 41 antibiotics. Gram-negative rods relative amount among pathogens decreased substantially (84.7 per cent in 1986 against 61.6 per cent in 1999), particularly Enterobacteriaceae (74.7 per cent in 1986 against 41.4 per cent in 1999). Nonfermenting Gram-negative rods (NFGNR) relative amount increased (10.8 per cent against 19.2 per cent), along with Gram-positive cocci (19.8 per cent against 64.2 per cent), particularly coagulasenegative staphylococci (CNS) (10.8 per cent against 35.9 per cent) and enterococci (5 per cent against 16.5 per cent) and candida and fungi (0.5 per cent in 1986 against 15.9 per cent in 1999). At the period 1986-1999 the main pathogens in urology infection were E. coli, Enterobacter spp., NFGNR (including P. aeruginosa), Staphylococcus, CNS, Enterococcus spp. The problem pathogens for urological department were the following: E. coli, Klebsiella spp., Enterobacter spp., Proteus spp., NFGNR including P. aeruginosa, CNS, Enterococcus spp., candida and fungi. At the period 1991-1997 Gram-negative pathogens susceptibility to amikacin, ofloxacin, ciprofloxacin, imipenem, ceftazidime, cefotaxime was not changed in general, Gram-positive cocci (staphylococci and enterococci) retained the same susceptibility to vancomicin, cefamandol and amoxyclave. Staphylococci were also susceptible to amikacin, imipenem, rifampicin, oxacillin, ciprofloxacin, and ofloxacin. Production of beta-lactamase was registered for 38.7 per cent of CNS, 26.5 per cent of E. coli, 38.5 per cent of K. pneumoniae, 25 per cent of P. mirabilis and 55.6 per cent of P. aeruginosa strains.

[Microflora in ENT and CNS diseases in patients living in the Moscow region]. / Mikroflora pri zabolevaniiakh Lor-organov i nervnoi sistemy u bol'nykh regiona Moskovskoi oblasti.

Patients with ENT and CNS inflammation living in the Moscow Region have been examined for pathogenic contamination of relevant clinical samples. The microflora changes were followed up, pathogenic agents of high and moderate priority were identified. It is shown that for Moscow Region, ENT and CNS purulent inflammation is associated primarily with coagulase-negative staphylococci, E. coli, meningococci, S. viridans and yeast fungi.