[Intravitreal aflibercept in high retinal pigment epithelial detachments]. / B L.N., Kh Iu.Iu., Z A.S., B A.S., K N.V. Intravitreal'noe vvedenie Aflibertsepta pri vysokikh otsloikakh retinal'nogo pigmentnogo épiteliia.

PURPOSE: To study the effectiveness of intravitreal aflibercept in high pigment epithelium detachment (PED) in eyes with age-related macular degeneration (AMD). MATERIAL AND METHODS: This study included 16 eyes of 16 patients (10 female and 6 male) aged 52 to 82 years (mean age 68.56±2.31 years). Examination of patients included determination of best-corrected visual acuity (BCVA), biomicroscopy with Goldmann lens, photographic recording of the ocular fundus, optical coherence tomography (OCT) and OCT-angiography. Results of the treatment were evaluated 1 month after the third injection of aflibercept. Among the analyzed parameters were: BCVA, maximum PED height, resorption of perifocal neuroepithelium detachment (NED), expressiveness of hyperreflective content of PED. RESULTS: One months into the treatment, after the third injection, mean BCVA improved by 0.09 to 0.36±0.04, height of PED decreased in average by 225.3±44.6 µm to 326.88±25.13 µm. Complete resolution of PED was not achieved in any of the cases. Resulting mean height of perifocal PED was 61.25±12.87 µm. Full resorption of PED was achieved in 4 eyes. Expressiveness of hyperreflective content of PED higher than 50% was observed in 13 eyes (81.25%). CONCLUSION: The results of intravitreal aflibercept in AMD patients with high PED have proved its effectiveness in reducing maximum PED height, improving resorption of perifocal NED and BCVA. Optically 'empty' PEDs had higher sensitivity to intravitreal aflibercept than hyperreflective PEDs. Statistically significant dependence of BCVA on the height and content of PED was absent.

Effect of Mifepristone on Corticosteroid Production in Vitro by Adrenal Glands of Rats with Streptozotocin Diabetes.

Changes in pregnenolone and corticosterone production by the adrenal glands of normoglycemic rats receiving the course of intraperitoneal mifepristone or NaCl administration were co-directed, but differed in magnitude. In rats with hyperglycemia, corticosteroid production increased after NaCl administration over 5 days and returned to the initial values after 15-day administration. On the contrary, pregnenolone and corticosterone production was suppressed after 5-day course of mifepristone, but significantly increased after mifepristone administration for 15 days. Intraperitoneal mifepristone administration almost completely abolished the response of rat adrenal glands with normo- and hyperglycemia to ACTH in vitro; this effect did not depend on the duration of administration.

Effects of Intraperitoneal Administration of Mifepristone on Glucocorticoid Status of Experimental Animals.

We studied the content of corticosterone and its precursors in the adrenal glands, corticosterone in blood serum and daily urine of rats, and activity of first and second isoforms of 11ß-hydroxysteroid dehydrogenase in the liver and kidneys of rats after 15 daily intraperitoneal injections of 0.9% NaCl or glucocorticoid receptor blocker mifepristone in 0.9% NaCl. Daily injections of NaCl reduced the levels of pregnenolone, progesterone, and corticosterone in the adrenal glands, increased corticosterone excretion with urine, enhanced activity of the first isoform of 11ß-hydroxysteroid dehydrogenase in the liver and reduction in activity of the second isoform of this enzyme in the kidneys. These changes are typical manifestations of chronic stress. Mifepristone restored pregnenolone content in the adrenal glands and increase in corticosterone concentration in the blood. Under these conditions, activity of the first isoform of 11ß-hydroxysteroid dehydrogenase in the liver did not change, and a decrease in activity of the second isoform of the enzyme in the kidneys was less pronounced. The results suggest that mifepristone abolished the stress-mediated increase in activity of the first isoform of 11ß-hydroxysteroid dehydrogenase in the liver and reduced local production of glucocorticoid hormones and their metabolic effects in hepatocytes.

[Diversity of the Key Biphenyl Destruction Genes in the Microbial Community of the Anadyr Bay Coastal Sediments].

Biphenyl 2,3-dioxygenase is the key enzyme involved in the bacterial destruction of biphenyl and polychlo- rinated biphenyls (PCBs), which are highly stable toxic compounds. The diversity of bphA1 genes encoding the biphenyl 2,3-dioxygenase a subunit of biphenyl-decomposing bacteria from the microbial community of the Bering Sea coastal sediments (the Anadyr port area) was studied. The enrichment culture was obtained by the incubation of bottom sediments samples with biphenyl as the only carbon source. It was followed by total DNA extraction and PCR analysis with degenerate primers specific to the bacterial biphenyl 2,3-dioxygenase a subunit genes. Subsequent cloning of the PCR products led to the identification of three types of aromatic dioxygenase genes, which appeared to be phylogenetically close to the genes of the biphenyl/toluene dioxygenase and 3-phenylpropionate dioxygenase subfamilies of the Actinomycetales bacteria.

[Comparative analysis of imipramine intake reactions in catatonic and wistar rats].

Chronic imipramine intake (7.5 mg/kg) leads to the stable decrease of excitable reactions to sound stimulant in Wistar rats and induced phase behavioral response in catatonic GC rats. Increased noradrenaline level in frontal cortex and striatum in Wistar animals was shown, whereas it didn't reveal noradrenaline level differences in any brain structures in GC rats. The higher blood corticosterone concentration was decreased under antidepressant reaction in GC rats. Differences between intact GC animals were found out: reduced triglyceride level, lesser body weight, and greater weight index of adrenals in comparison with Wistar rats. Various mechanisms of imipramine action in two rat strains were shown: influence on adrenergic brain system, taking part in the control of exiting behavior in Wistar rats and reaction in adrenals in GC rats.

Activity of 11ß-hydroxysteroid dehydrogenase in the adrenal glands, liver, and kidneys of rats with experimental diabetes.

We studied activity of the key enzyme of the pre-receptor metabolism of glucocorticoid hormones, 11ß-hydroxysteroid dehydrogenase, in rat adrenal glands, renal cortex and liver in the course of development of alloxan diabetes (9, 20, and 28 day). The enzyme activity was increased 3-4 fold in the adrenal glands throughout the experiment. At the same time, according to the adrenal gland level of corticosterone, its precursor 11-deoxycorticosterone and reversible metabolite 11-dehydrocorticosterone, activity of the second isoform of the enzyme dominated at the early stages of diabetes, and that of the first isoform, at later stages. In long-term diabetes (28 days), along with reduced synthesis of corticosterone and production of 11-dehydrocorticosterone in the adrenal glands, the extra-adrenal formation of corticosterone was activated as indicated by enhanced activity of the first isoform in the liver and that of the second isoform in the kidneys. These changes in activity of the enzyme isoforms promote local formation of corticosterone from its reversible metabolite in the liver and persisting hyperglycemia in diabetes.

Production of corticosteroid hormones in vitro by adrenals in rats with streptozotocin-induced diabetes.

We studied baseline and ACTH-stimulated in vitro production of corticosteroids by rat adrenals. Production of the basic corticosteroids pregnenolone (early precursor in corticosteroid synthesis), progesterone (intermediate precursor in synthesis of gluco- and mineralocorticoid hormones), and corticosterone (major glucocorticoid hormone in rodents) in animals with streptozotocin-induced diabetes was enhanced by 1.8-2.0 times in comparison with the control animals. Addition of ACTH to the incubation medium stimulated pregnenolone production by the adrenals equally in the control and experimental (diabetic) groups, while the increase in corticosterone production was less pronounced in the experimental group. Stimulation of corticosterone production in response to ACTH after saturation of the incubation medium with pregnenolone was also less pronounced in diabetic rats.

[Photosensitizing properties of 3,3'-diethylthiacarbocyanine in biological media].

The objective of the study is elucidation of perspectives of 3,3'-diathylcarbocyaine application as a photosensitizer for curing viral infections by photodynamic therapy. Lipid-containing bacteriophage PM-2 of Pseudoalteromonas espejiana was used as a model. The testing was carried out at a special installation modeling photodynamic exposure conditions towards a non-fractionated phage lysate. 3,3'-DECC demonstrated a rapid photo-bleaching when added tothe phage lysate but not to water. The initial rate of PM-2 phage photoinactivation was proportional to the square concentration of the dye in the range of 0.5-9 µmol/L. This confirms a hypothesis that the dimer is the principal photochemically active form of the dye. An improved ability to form dimers was found in the dye in the phage lysate (10-folds better than in the water). The dye formed a stable adduct with the bacteriophage material. This adduct had an extinction maximum at λ(max) = 594 nm and demonstrated the properties of a polymer (sedimentation under a low-speed centrifugation).

Effect of mifepristone on glucocorticoid receptor gene expression in the liver of rats with streptozotocin-induced diabetes.

The effects of mifepristone on activity of the adrenocortical system, expression of glucocorticoid receptor gene, and tyrosine aminotransferase activity in the liver were studied in rats with streptozotocin-induced diabetes. Administration of glucocorticoid receptor blocker mifepristone to rats without diabetes was followed by a 1.9-fold increase in serum corticosterone concentration and a 1.2-fold increase in tyrosine aminotransferase activity in the liver in comparison with the baseline values. In rats with streptozotocin-induced diabetes, mifepristone produced a less pronounced increase in the corticosterone concentration (by 1.5 times) and more drastic increase in enzyme activity (by 1.7 times). Mifepristone administration did not change the content of glucocorticoid receptor mRNA in the liver of rats without diabetes, but increase this parameter by 1.4 times in rats with streptozotocin-induced diabetes. The enhanced expression of glucocorticoid receptor gene in the liver of rats with streptozotocin-induced diabetes correlated with increased activity of tyrosine aminotransferase after mifepristone treatment.

Prospects of using photostimulation of the visual analyzer in sports medicine.

Experimental studies of photostimulation of the visual analyzer by combinations of red, green, blue, and yellow colors in a pulsed mode were carried out. Significant improvement of a visual acuity is shown after photostimulation course in patients with macular degeneration and degenerative changes of the optic nerve.

Adrenocortical system response to induction of inflammation with silicon dioxide in rats with alloxan-induced diabetes mellitus.

Alloxan-induced diabetes mellitus in rats was characterized by persistent increase in blood levels of corticosterone, while chronic granulomatous inflammation induced by silicon dioxide and its combination with alloxan-induced diabetes mellitus were associated with transient increase in blood corticosterone level followed by gradual development of hypoadrenocorticism. The content of corticosterone in the adrenal glands of rats with alloxan-induced diabetes mellitus remained unchanged in the dynamics of the disease, but the level of progesterone decreased at the early terms of diabetes and then returned to the initial values. After administration of silicon dioxide to intact rats and to rats with diabetes mellitus, changes in hormone content in the adrenal glands were observed only at the initial stages of inflammation and consisted in elevation of corticosterone concentration against the background of reduced progesterone content.

Effect of imipramine on nerve excitability in GC rats.

Imipramine test (7.5 mg/kg) revealed a persistent positive reaction of Wistar rats, which manifested in reduced excitability of animals. Oral administration of imipramine solution was followed by unstable behavioral reactions in GC rats. Norepinephrine concentration in the cortical and limbic regions of these animals remained unchanged, while plasma corticosterone concentration decreased to the control level and did not differ from that in Wistar rats. Our results indicate that imipramine has a modulatory effect on destabilization of the adaptive system in catatonic GC rats.

[Application of the complex of DNA with the congo red anionic diazo dye for detection of nuclease-producing colonies of marine bacteria].

This study was aimed at the development of a method for detection of colonies of nuclease-secreting marine bacteria. The BAL nuclease-producing marine bacterium Pseudoalteromonas espejiana BAL-31 was used as the test object. A new method was developed involving the congo red (CR) anionic dye. The P. espejiana culture was plated on nutrient agar with CR and denatured DNA. In such media. CR was found to form complexes with DNA. After two days of incubation at 30 degrees C, halos were found around the P. espejiana colonies. No halos appeared when DNA was not introduced, when BAL nuclease was inactivated, or when the medium was inoculated with Escherichia coli. It was concluded that the halos around the colonies indicated nuclease excretion. The halos were shown to result from the coagulation of CR released after digestion of the CR-DNA complex by the nuclease. This method for detection of nuclease-producing colonies can probably be used for all marine bacteria and possibly for halophilic bacteria as well.

[A novel method of transfection of marine bacteria Pseudoalteromonas espejiana with deoxyribonucleic acid of bacteriophage PM2].

DNA of bacteriophage PM2 is a convenient test object for studying DNA-damaging genotoxic agents. The extent of DNA damage can be estimated by the ability of damaged DNA for transfection of host cells, marine bacterium Pseudoalteromonas espejiana (Pae), str. BAL-31. The efficiency of transfection of Pae lines maintained for long periods without freezing was found to be very low upon the use of a widely accepted transfection method developed by van der Schans et al. (1971). Such cultures grown in a medium with 10 mM Ca2+ standard for Pae contained cell aggregates and exopolymer material. Pae was found to be capable of growing in a medium without the calcium supplement in the presence of chelator EGTA (low-calcium medium, LCM). After growth in LCM, cells did not aggregate, cultures lacked the activity of nuclease BAL, and transfection efficiency of cells grown in LCM drastically increased. Based on these results, a novel procedure of transfection with an efficiency of 2 x 10(4)-2 x 10(5) infectious centers per microgram of PM2 DNA was developed.

[UHF-dielectrometry in the assessment of the structural organization of salt solutions and interstitial fluids in normal and cicatricial tissues]. / KVCh-diélektrometriia pri otsenke strukturnoi organizatsii rastvorov solei i tkanevykh zhidkostei zdorovoi i rubtsovoi tkani.

The complex dielectric permittivity of salt solutions with positive and negative salvation as well as healthy and cicatricially changed human skin in situ at the frequencies of 42 and 56.6 GHz was measured. The relation between the dielectric characteristics of water and diluted salt solutions and changes in their structural organization conditioned by different temperatures of samples and the type of salvation of electrolytes was studied. The differences in the dielectric characteristics of healthy and cicatricially changed skin are interpreted in terms of the dependence of the structural organization of interstitial fluids on the morphological and functional state of biological tissues.

[Photoinactivation of bacteriophage PM2 by cyanine dyes]. / Fotoinaktivatsiia bakteriofaga PM2 v prisutstvii tsianinovykh krasitelei.

Photoinactivation of the lipid-containing bacteriophage PM2 by visible light and cyanine dyes (carbo- and dicarbocyanines), aluminum phtalocyanine tetrasulfonate and methylene blue was studied. It was concluded that cyanine dye aggregates adsorbed on phage particles and oxygen are essential for phage photoinactivation.

[Sensitivity of Russian HIV-1 isolates to anti-HIV agents]. / Chuvstvitel'nost' k anti-VICh-preparatam izoliatov VICh-1, vydelennykh na territorii Rossii.

HIV strains isolated from HIV patients hospitalized at the Central Research Institute of Epidemiology in 1991-1993 are analyzed. HIV-1 strains isolated at different stages of the illness were referred to subtypes A and B. The biological properties of the isolates of both subtypes were virtually the same. The majority of the isolated HIV-1 strains were relatively resistant to azidothymidin, no matter whether the patients were previously treated with this drug or not. Study of strains repeatedly isolated from the same patients showed an increase of the cytodestructive characteristics of the isolates. Strains isolated from the same epidemiological chain were characterized by the same properties and sensitivity to the tested antiHIV agents. The resistance of the virus to antiHIV drugs with different mechanisms of action may be an indicator of the terminal stage of the illness.

Comparative study of thermostability and structure of close homologues--barnase and binase.

Parameters of heat denaturation and intrinsic fluorescence of barnase and its close homologue, binase in the pH region 2-6 have been determined. The barnase heat denaturation (pH 2.8-5.5) proceeds according to the "all-or-none" principle. Barnase denaturation temperature is lower than that of binase and this difference increases from 2.5 degrees C at pH 5 to 7 degrees C at pH 3. Enthalpy values of barnase and binase denaturation coincide only at pH 4.5-5.5, but as far as pH decreases the barnase denaturation enthalpy decreases significantly and in this respect it differs from binase. The fluorescence and CD techniques do not reveal any distinctions in the local environment of aromatic residues in the two proteins, and the obtained difference in the parameters of intrinsic fluorescence is due to fluorescence quenching of the barnase Trp94 by the His 18 residue, absent in binase. Secondary structures of both native and denaturated proteins also do not differ. Some differences in the barnase and binase electrostatic characteristics, revealed in the character of the dipole moments distribution, have been found.

[Comparison of the heat stability and structure close homologs--Bacillus amyloliquefaciens ribonuclease and Bacillus intermedius 7P ribonuclease]. / Sravneniee termostabil'nosti i struktury blizkikh gomologov-- ribonukleazy Bacillus amyloliquefaciens i ribonukleazy Bacillus intermedius 7P.

Parameters of heat denaturation and intrinsic fluorescence of barnase and its close homologue, binase, in the pH region 2-6 have been determined. Barnase heat denaturation (pH 2.8-5.5) proceeds according to the "all-or-none" principle. Barnase denaturation temperature is lower than that of binase and this difference increases from 2.5 degrees C at pH 5 to 7 degrees C at pH 3. Enthalpy values of barnase and binase denaturation coincide only at pH 4.5-5.5, but as the pH decreases the barnase denaturation enthalpy decreases significantly and in this respect it differs from binase. The fluorescence and CD techniques do not reveal any distinctions in the local environment of aromatic residues in the two proteins, and the obtained difference in the parameters of intrinsic fluorescence is due to fluorescence quenching of the barnase Trp-94 by the His-18 residue, which is absent in binase. Secondary structures of both native and denaturated proteins also do not differ. Some differences have been found in the barnase and binase electrostatic characteristics, revealed in the character of the dipole moment distribution.