[Testing of the Russian-language version of the Thought, Language and Communication Scale]. / Aprobatsiya russkoyazychnogo varianta shkaly Thought, Language, Communication Scale.

OBJECTIVE: To analyze the possibilities of using the Russian version of the Thought, Language and Communication Scale (TLC) for early recognition of thought disorders in patients at clinical high-risk for schizophrenia. MATERIAL AND METHODS: For the main group, the study included 30 adolescent male patients (19.2±2.2 years) hospitalized with the first depressive episode (ICD-10: F32.1, F32.2, F32.28, F32.8), who demonstrated attenuated schizophrenic symptoms (ASS) in the structure of the depression, which made it possible to attribute the patients to the group of clinical high-risk for schizophrenia. The control group consisted of 27 mentally healthy adolescent males (20.0±2.3 years). In both groups, the severity of thought impairment was assessed using the TLC scale. Psychopathological, psychometric and statistical methods were used. RESULTS: The median values of the severity of thought impairment using the TLC scale were 20 points [19.75; 26] in the main group, 10.5 points [9.25; 13] in the control group, with a high degree of statistical significance (p<0.001). The most significant differences (p<0.001) were found in following parameters: Incoherence (2 [1; 3] vs 1 [0; 1]), Tangentiality (2 [2; 2] vs 1 [0; 2]), Derailment (2 [1.25; 2] vs 1 [0.5; 2]), Illogical thinking (2 [2; 2.75] vs 0 [0; 1]), Loss of goal (1 [0; 2] vs 0 [0; 0]) and Blocking (1 [0; 1] vs 0 [0; 0] accordingly). CONCLUSION: Specific, not related to depression, disorders of thinking in patients of the clinical group, which indicates signs of disorganization of thinking and suggests the beginning of the endogenous process of the schizophrenic pole were found. The results show that the TLC scale can be used to detect early cognitive disorders in patients at risk of schizophrenia.

[The relation to the humor and laugh in patients with schizophrenia].

This study provides the data on gelotophobia (the fear of being laughed at), gelotophilia (the joy of being laughed at) and katagelasticism (the joy of laughing at others) in patients with schizophrenia in comparison with controls. Gelotophobia was significantly higher and katagelasticism was significantly lower in psychiatric patients than in controls. The negative correlation was found between gelotophobia and the duration of illness among patients with paranoid schizophrenia. Gelotophilia and katagelasticism were correlated positively both in patients and controls. In patients with schizotypal disorder, gelotophobia was also correlated with katagelasticism. High scores on katagelasticism and gelotophilia typical for men in the control group became typical for women in schizophrenia group.

[Amino acids content in the blood serum of patients with multiple sclerosis].

Content of amino acids in the blood serum using ion-exchange chromatography and a quantitative crystalloscopic "closed drop" method was measured in 105 patients with multiple sclerosis of different severity. The relation between the dynamics of content of glutamic, asparagine acids and glycine and the disease severity was determined. The highest level of glutamic acid was found in the patients with severe disease course. The dynamics of quantitative changes in the amino acid content was accompanied by the disturbance of blood serum structuration and appearance of selective micromorphotypes. The positive correlation between the severity of motor and coordination disorders and the increase in levels of glutamic acid and glycine in the severe cases was detected.

Bridged oligonucleotides as molecular probes for investigation of enzyme-substrate interaction and allele-specific analysis of DNA.

The efficiency of enzymatic conversion of DNA complexes containing non-nucleotide inserts has been studied. T4 DNA ligase and Taq DNA polymerase have been included in the study as examples of widely used DNA-dependent enzymes. A series of substrate DNA complexes have been formed using native oligonucleotides and bridged ones bearing non-nucleotide inserts based on phosphodiesters of di-, tetra-, or hexaethylene glycol, 1,5-pentanediol, 1,10-decanediol, and 3-hydroxy-2(hydroxymethyl)-tetrahydrofuran. The perturbation in DNA located far from the site of the enzyme action had almost no influence on the substrate properties of the complex, while insertion near this site significantly deteriorated them. The use of a series of modified duplexes allows one to locate the position of the enzyme-binding site on DNA substrate with the accuracy of 1-2 nucleotides. The presence of a non-nucleotide insert in the complex has been also shown to enhance the efficiency of single mismatch discrimination upon both template-directed ligation and extension of oligonucleotides.

[Enhancement of hybridization analysis efficacy by means of controlled fragmentation of analyzed DNA].

The influence of fragmentation of analyzed DNA amplicon on the efficacy of specific sequence detection by means of heterophase hybridization analysis was investigated. The detection of DNA sequence was carried out colorimetrically after introduction of biotin label into the oligonucleotide probe immobilized on a solid support upon its limited elongation in the complex with the analyzed DNA using Taq polymerase. Two simple and reproducible approaches to DNA analyte fragmentation were suggested. They are based on the formation apurinic/apyrimidinic sites in DNA followed by their degradation upon the thermal treatment. Apurinization of DNA was achieved with a mild acidic treatment. The apyrimidinic sites were formed when DNA fragment containing dTMP and dUMP residues in various ratios was treated with uracil-DNA-glycosylase (UDG). DNA analytes pretreated by one of these approaches can be used without additional purification for hybridization analysis of DNA with the use of Taq polymerase. The efficacy of hybridization analysis is shown to be higher in the case of the fragmented DNA in comparison with the native DNA amplicon. The use of fragmented DNA analytes allows utilizing bridged oligonucleotides as highly selective probes with reduced hybridization properties.

Hybridization of the bridged oligonucleotides with DNA: thermodynamic and kinetic studies.

Hybridization properties of oligonucleotides containing non-nucleotide inserts designed on the basis of synthetic abasic sites, oligomethylene diols or oligoethylene glycols have been characterized. The influence of the inserts which generate extrahelical anucleotidic bulges on thermodynamics, kinetics of hybridization of bridged oligonucleotide with DNA has been studied by UV-melting and stopped-flow techniques. Circular dichroism spectrometry data show that anucleotidic bulges in the middle of the duplex does not alter the B-form helix conformation. Nevertheless, the insert induces destabilization of the duplex structure, caused mostly by the considerable enhancement of the dissociation rates. Free energy increments for the extrahelical anucleotidic bulges can be described in the nearest-neighbor approximation. The thermodynamic effect of the insert lengthening obeys a simple Jacobson-Stockmayer entropy extrapolation. Independently of the insert type, the free energy term is directly proportional to the logarithm of the number of bonds between the oligonucleotide fragments. The behavior of hydrophobic inserts formed by 10-hydroxydecyl-1-phospate units is an exception to the rule.

[Universal method for single nucleotide substitution identification].

A new approach to the identification of point mutations by allele-specific PCR was proposed. The mutation R408W of the human phenylalanine hydroxylase gene was used as a model. A high specificity of the approach was achieved by the use of primers partially complementary to the genomic DNA. Polyethylene glycol covalently attached to one of the allele-specific primers provides for the differential identification of the PCR products due to a change in electrophoretic mobility.

Ribonuclease activity of the peptides with alternating arginine and leucine residues conjugated to tetrathymidilate.

RNA cleaving conjugates have been prepared by attachment of oligodeoxyribonucleotide TTTT to peptides containing arginine, leucine, proline and serine residues. The highest activity was displayed by the conjugates containing peptides with alternating arginine and leucine residues (LR)4G-amide. Ribonuclease activity of the conjugates pep-T4 decreases in the order T4-(LR)4G > T4-(LR)2G > T4-(LLRR)2G > T4-(LR)2PRLRG > S2R3-Hmda-T4 > or = R5 double dagger (LR)3. According to CD spectra, the free peptide (LR)4G-amide in water solution at neutral pH and physiological ionic strength has no pronounced secondary structure whereas conjugated to oligonucleotide it acquires a folding similar to alpha-helix.

[Detection of the DeltaF508 trinucleotide deletion in the human CFTR gene by UV immobilization on nylon of a complex of a PCR fragment with a highly specific probe]. / Vyiavlenie trinukleotidnoi deletsii DeltaF508 v gene CFTR cheloveka putem UF-immobilizasii na kaprone kompleksa PTsR-fragmenta s vysokospetsifichnym zondom.

Deletion delta F508 has been revealed in PCR-amplified regions of human gene CFTR by color detection of the hybridization complex obtained by ligation of a tandem of short oligonucleotides on a DNA template followed by UV immobilization on nylon. The method allows reliable detection of the three-nucleotide deletion (insertion). The nonspecific signal depends on the nucleotide composition of the biotinylated tandem component. A significant level of the specific signal was achieved by using the PCR-amplified DNA fragments of different length (200-400 bp) irrespective of the position of the tandem-binding site in their sequences.

[A new approach to detect a particular DNA sequence by UV-immobilization of its hybridization complex with a highly specific probe resulting from ligation of a tandem of short oligonucleotides in solution]. / Novyi podkhod k vyiavleniiu analiziruemoi posledovatel'nosti DNK putem UF-immobilizatsii ee gibridizatsionnogo kompleksa s vysokospetsifichnym zondom, obrazuiushchimsia pri ligirovanii tandema korotkikh oligonukleotidov v rastvore.

A new approach was proposed for detecting amplified DNA fragments by hybridization with a highly selective oligonucleotide probe obtained by ligation of a tandem of three short oligonucleotides (pN8 + pN4 + pN8' Bio) in solution, with subsequent UV-immobilization of the hybridization product on a nylon membrane and its colorimetric detection with the streptavidin-alkaline phosphatase technique. Owing to the high selectivity of ligation, the 20-mer ligation product was detected on a membrane only when it was completely complementary to a template fragment. The results showed that any single-nucleotide substitution in the tetramer-binding site can be localized and identified with the use of all 12 possible tetramers.

Nuclease resistance and RNase H sensitivity of oligonucleotides bridged by oligomethylenediol and oligoethylene glycol linkers.

The properties of new chimeric oligodeoxynucleotides made of short sequences (tetramers, pentamers, octamers, and decamers) bridged by hexamethylenediol and hexaethylene glycol linkers have been investigated. These chimeric oligonucleotides showed an improved resistance toward snake venom 3'-phosphodiesterase, with an increased stability when a terminal 3'-3'-internucleotide phosphodiester bond is present. It also has been demonstrated that the hybrid complexes formed by bridged oligonucleotides and a complementary 20-mer RNA are able to elicit the activity of ribonuclease H (RNase H) from Escherichia coli. The substrate properties of chimeric oligonucleotides depend on the length of the oligonucleotide fragments bridged by linkers. Introduction of a nonnucleotide spacer into the native oligonucleotide only slightly hampers the extent of the RNA hydrolysis in the hybrid complexes, whereas a modification of the site of reaction is observed as a possible consequence of the steric disturbance due to the aliphatic linkers. Hence, these new chimeric oligonucleotides, namely, short oligonucleotide fragments bridged by nonnucleotide linkers, demonstrate a favorable combination of exonuclease resistance and high substrate activity toward RNase H. As a consequence, these chimeric oligonucleotides could be proposed as new, promising analogs to be used in the antisense strategy.

[The nature of stabilization of the tandem DNA duplex pTGGAGCTG (pCAGC + (Phn-NH-(CH2)3-NH)pTCCA) based on the UV-, CD-, and two-dimensional NMR spectroscopy data]. / Priroda stabilizatsii tandemnogo DNK-dupleksa pTGGAGCTG (pCAGC + (Phn -NH-(CH2)3-NH)pTCCA) po dannym UF-, KD- i dvumernoi IaMR-spektroskopii.

Properties and three-dimensional structure of the tandem DNA duplex pTGGAGCTG.(pCAGC+(PhnL)pTCCA) in aqueous solution, where L is an amino linker and Phn is an N-(2-hydroxyethyl)phenazinium residue, were studied spectrophotometrically and by two-dimensional 1H NMR spectroscopy (COSY and NOESY). When a tandem complex involving a Phn residue-bearing oligonucleotide is formed, the dye aromatic system intercalates into the double helix at the nick site and takes part in two stacking interactions: a strong one (3.5-4 A) with the T5-A12 base pair of its own duplex moiety and a weak one (4-5 A) with the C4-G13 pair of the adjoining duplex (mainly with the C4 base). This arrangement of the dye residue, providing a cross-interaction of the phenazinium moiety with the base pairs of the adjacent duplex structures, results in the stabilization of the whole tandem complex.

A new strategy of discrimination of a point mutation by tandem of short oligonucleotides.

A new strategy based on the use of cooperative tandems of short oligonucleotide derivatives (TSOD) has been proposed to discriminate a "right" DNA target from a target containing a single nucleotide discrepancy. Modification of a DNA target by oligodeoxyribonucleotide reagents was used to characterize their interaction in the perfect and mismatched complexes. It is possible to detect any nucleotide changes in the binding sites of the target with the short oligonucleotide reagent. In the presence of flanking di-3',5'-N-(2-hydroxyethyl)phenazinium derivatives of short oligonucleotides (effectors) the tetranucleotide alkylating reagent modifies DNA target efficiently and site-specifically only in the perfect complex and practically does not modify it in the mismatched complex. It has been shown that TSOD is much more sensitive tool for the detection of a point mutation in DNA as compared to a longer oligonucleotides.

[Cleavage of RNA in hybrid duplexes by E. coli ribonuclease H. II. Substrate properties of nucleotides containing non-nucleotide linkers]. / Rasshcheplenie RNK v sostave gibridnykh dupleksov ribonukleazoi H E. coli. II. Substratnye svoistva oligonukleotidov, soderzhashchikh nenukleotidnye vstavki.

The 20-mer bridged oligodeoxynucleotides containing short oligomers joined by the hexamethylenediol and hexaethylene glycol linkers were shown to form complementary DNA/DNA and RNA/DNA complexes whose thermostability depends on the length and number of the nonnucleotide linkers. Hybrid complexes of the bridged oligonucleotides proved to be substrates for the E. coli ribonuclease H. The presence of one-three nonnucleotide linkers in a 20-mer decreased the hydrolysis efficacy only 1.2-1.4-fold. It is the composition of the RNA cleavage products that was influenced the most significantly by the nonnucleotide linkers. RNase H simultaneously hydrolyzed the RNA 3'-ends of each hybrid duplex involving a bridged oligonucleotide. The presence of an inverted 3'-3'-phosphodiester bond at the 3'-end of the oligodeoxyribonucleotide only slightly affected the RNase H activity.

[Possibility of correction of erythrocyte fatty acid composition in men with anamnesis of hereditary ischemic heart disease]. / Vozmozhnost' korrektsii sostava zhirnykh kislot eritrotsitov u muzhchin s otiagoshchennym anamnezom po ishemicheskoi bolezni serdtsa.

The results of 4-year monitoring of men with hereditary history of ischemic heart disease are presented. The examinees were divided into two groups: a group of diet correction with alimentary omega-3 polyunsaturated fatty acid (PUFA) extracted from Far Eastern sardine oil and a control group which did not take part in the preventive treatment. The composition of fatty acids of erythrocytes of men from the study and control groups was analyzed in the course of preventative treatment, 3, 6 months and 4 years after it. It has been proved that the progress of disorder in composition of fatty acids in men can be stopped by using alimentary omega-3 PUFA.

[Interaction of derivatives of short oligonucleotides with nucleic acids. VIII. Characteristics of target DNA modification by alkylating oligonucleotide derivatives in tandem complexes]. / Vzaimodeistvie proizvodnykh korotkikh oligonukleotidov s nukleinovymi kislotami. VIII. Osobennosti modifikatsii DNK-misheni v tandemnykh kompleksakh alkiliruiushchimi proizvodnymi oligonukleotidov.

The influence of effectors [octanucleotides and their 3',5'-di-N-(2-hydroxyethyl)phenazinium derivatives] on the modification of a target DNA by alkylating oligonucleotide derivatives forming duplexes of different stability with the target ws studied. It is shown that, being in tandem complexes immediately adjacent to the reactive group of an oligonucleotide reagent possessing a high hybridization capacity, the effector, on the one hand, enhances the stability of the reagent target duplex, and on the other hand, changes the site-specificity of alkylation and decreases the efficiency of the target modification at temperatures that provide a high extent of the target association with the reagent. Conversely, in the case of oligonucleotide reagents forming weak complexes with the target, effectors enhance both the stability of the target.reagent duplex and the extent of the target throughout the temperature range tested. The data indicate that the varying influence of effectors on the target modification by reagents with different hybridization capacities is due to conformational features of the target reagent duplexed regions. Increasing the rigidity of the target.reagent duplex reduces the efficiency of the target modification in tandem complexes.