[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.

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.

[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.

[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.

[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.

[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.

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.

[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.

Subcellular distribution of the R-subunits of cAMP-dependent protein kinase in LS-174T human colon carcinoma cells.

The analysis of the particulate preparations of LS-174T human colon carcinoma cells in confluent stage of growth revealed different distribution for regulatory subunits (R) of cAMP-dependent protein kinase (PKA) in the subcellular compartments. The LS-174T cell lysates were subjected to differential and discontinuous sucrose density gradient centrifugation. The obtained fractions were assayed for marker enzymes and photoaffinity labeled with 8-N3[32P]cAMP. The whole lysates and cytoplasmic fraction exhibited the presence of both RI alpha and RII alpha--subunits of PKA. The fractions exhibiting high activity of the marker enzymes for plasma membranes, Golgi apparatus and mitochondria contained mainly RII alpha. In the fractions of lysosomes and microsomes RI alpha and RII alpha were found in nearly equal amounts.

Chemical modification enhances the inhibitory effect of regulatory subunit antisense oligodeoxynucleotide of cAMP-dependent protein kinase type I on cell proliferation.

The analysis of the action of antisense oligodeoxynucleotide complementary to mRNA of the type I cAMP-dependent protein kinase regulatory subunit has revealed a stable inhibitory effect of the substance on Molt-4 leukemic cell proliferation. To enhance the efficiency of the oligodeoxynucleotide action, its molecule was subjected to terminal modifications. The hydrophobic radical-effected modification at the 5'-end was carried out at the last stage of automated synthesis using undecanol. At the 3'-end, the molecule was modified with an acridine residue. The substances synthesized were significantly more potent in inhibiting thymidine incorporation in the acid-insoluble fraction, compared to the nonmodified nucleotide. Examination of the cell cycle progression of Molt-4 cells has demonstrated a considerable shift of cells from G2/M stage to GI, the process being more effective in the case of modified oligonucleotides.

[Introduction of regulatory subunits of cAMP-dependent protein kinase type II in ZTZ cells with the use of erythrocyte ghosts]. / Vvedenie reguliatornoi sub''edinitsy cAMP-zavisimoi proteinkinazy II tipa v kletki ZTZ s primeneniem tenei éritrotsitov.

The dynamics of distribution of the regulatory subunit of cAMP-dependent protein kinase II following protein injection into 3T3 cells was studied. The cAMP-binding component of protein kinase was injected into the cells, using erythrocyte ghosts. The conditions for protein encapsulation into erythrocyte ghosts were elaborated. The optimal detergent concentrations, incubation time and conditions of vesicular closure following protein injection were selected. The above method provides for a high (50-55%) yield of the erythrocyte ghost-encapsulated protein with a minimum loss of enzymatic activity. Fusion of erythrocyte ghosts containing the labeled protein with 3T3 cells was carried out. Using the cytoradiography technique, the dynamics of distribution of the radiolabeled regulatory subunit within the cell was analyzed. It was demonstrated that after the regulatory subunit has reached the cytoplasm, the protein is translocated into the nucleus and is pooled there is the vicinity of the nucleoli.

[Mechanisms of the conduction of cellular regulatory signals]. / Mekhanizmy provedeniia reguliatornogo signala v kletku.

A rat isolated heart was used to demonstrate a protective effect of phosphocreatine (10 mM) in total normothermic ischemia (25 min) with subsequent reperfusion with the Krebs-Henzeleit solution and in cardioplegia in the St Thomas Hospital solution. The effect consisted in reduction in ischemic and reperfusion contraction and a 2-3-fold increase in cardiac output as compared with the control. A 20 per cent increase in the calcium content in the solution did not influence the phosphocreatine effect. Application of tocopheryl phosphate (0.1 microM) had no effect on the ischemic contraction but reduced diastolic pressure on reperfusion and provided a better maintenance of the cardiac parameters. The tocopheryl phosphate action increased when used in combination with phosphocreatine. The findings show that phosphocreatine slows down the development of ischemic damage and onset of irreversibility while antioxidants exert a protective effect by preventing a reperfusion damage to the heart.

Dynamic distribution of the regulatory subunit of cAMP-dependent protein kinase type II in NIH 3T3 cells.

Experiments on the introduction of the regulatory subunit of cAMP-dependent protein kinase type II (RII) into NIH 3T3 cells clearly demonstrated its translocation into the nucleus. The labelled protein was incorporated into erythrocyte ghosts and their fusion with the cells was carried out. The dynamics of distribution of the labelled RII in NIH 3T3 cells was studied by the method of historadiography. It was found that during the next few hours after its penetration into the cytoplasm, the protein translocates into the nucleus and concentrates in the immediate proximity to the nucleoli.