[Pharmacogenetic aspects of the dopaminergic system in clozapine pharmacodynamics]. / Farmakogeneticheskie aspekty dofaminergicheskoi sistemy v farmakodinamike klozapina.

A systematic review of association studies on the role of single nucleotide variants (SNVs) of the dopaminergic system genes on the effectiveness of clozapine in schizophrenia has been perfromed. A search of literature was conducted in PubMed, MedLine, Web of Science Core Collection (Clarivate Analytics), Web of Science, Russian Science Citation Index, Scopus, Scientific Research, Google Scholar, Oxford Press, e-Library from 1995-2019. Association studies of 53 SNPs of genes encoding dopamine receptor isoforms (DRD1), dopamine transporter (SCL6A3) and catechol-O- methyltransferase (COMT), and the nature of their association with the therapeutic response to clozapine were analyzed. The results of SNPs studies of DRD1 and COMT genes are the most controversial. This can be explained by the heterogeneity of the samples and the lack of standardization of methods for evaluating the effectiveness of treatment in the context of association studies. The clear population specificity of the association of some SNPs of DRD1, DRD2 and DRD3 genes with the response to clozapine therapy has been shown. Most of the identified associations are haplotype specific. The obtained regularities of the effect of SNPs of dopaminergic system genes on the effectiveness of clozapine therapy should be considered in an individual approach to treatment of schizophrenia.

[New morphometric criteria in the study of pathogenesis of normal-tension glaucoma]. / Novye morfometricheskie kriterii v izuchenii patogeneza glaukomy normal'nogo davleniya.

INTRODUCTION: Studying the factors that contribute to the disturbance of transmembrane pressure gradient is a topical task in the research of pathogenesis of normal-tension glaucoma (NTG). PURPOSE: To measure and compare the thickness and depth of lamina cribrosa (LCT and LCD), as well as optic nerve subarachnoid space width (ONSASW) in patients with NTG and healthy individuals. MATERIAL AND METHODS: The first group included 12 patients (23 eyes) aged from 58 to 74 years (average age 66.8±3.2 years) with NTG who all had normal intraocular pressure and arterial hypotension. The second (control) group consisted of 11 healthy individuals (22 eyes) aged from 51 to 69 years (average age 56.2±4.2 years). All patients underwent structural and functional assessment of the optic nerve head using OCT RTVue-100 (Optovue, USA), as well as Humphrey Visual Field Analyzer (HFA II 745i, Germany-USA) and our modification of Frequency Doubling Technology perimetry. In all subjects the central corneal thickness (CCT) was measured by Pentacam HR (Oculus, Germany), the LCT and LCD were measured by EDI (Enhanced Depth Imaging) mode of Topcon 3D OCT 2000 (Japan). To measure the ONSASW we used a cross-sectional image of the optic nerve taken 3 mm behind the eyeball with Magnetic Resonance Imaging (Siemens Magnetom Symphony 1.5 T, Germany). RESULTS: Statistically significant difference was found between the 1st and 2nd groups in the average LCT (217.60±36.92 and 345.86±33.29 µm respectively; p=0.0000), LCD (435.00±86.31 and 367.31±87.00 µm, respectively; p=0.014) and ONSASW (1.27±0.13 and 1.44±0.19 mm respectively; p=0.004); the difference wasn't significant in the average CCT (543.26±31.52 and 557.50±24.92 µm respectively; p=0.11). CONCLUSION: Patients with NTG had significantly higher value of the LCD with significantly lower values of the LCT and ONSASW compared with healthy individuals, which confirms the importance of these morphometric criteria in the study of NTG pathogenesis.

[The Spatial Variability of Methane Emission from Grass-Moss Fens of the Subtaiga and Forest-Steppe of Western Siberia].

Emission of methane from the grass-moss fens of the Western Siberia subtaiga was studied using a static chamber method. It was established that a median half of the interquartile range of the specific flow of CH4 in the studied wetland ecosystems constituted 4.9 ± 2.9 mg of CH4/(m2 x h). It was shown that such a high spatial variability of emission is caused mainly by the difference in the level of fen waters. It was found that, in these observations, a higher level of the water stand correlates with lower emission values. The causes of this phenomenon are discussed, and recommendations for conducting field studies for estimating the regional flow are given.

[Optimization of staged treatment in patients with extensive combined face defects].

Free autotransplantation of revascularized flaps allow reconstructing soft tissue integrity with simultaneous bone defects elimination in patients with extensive combined face defects, but for eligible reconstruction of facial proportions one may need additional surgical procedures. The study included 161 patients who underwent free autotransplantation of revascularized flaps. Main pitfalls of the method are revealed and reviewed in the paper. When corresponding to deformity type free autotransplantation of revascularized flaps results in full functional and esthetic rehabilitation.

[Modulation of rat myometrium contractile activity by peptidoglycan of Staphylococcus aureus cell wall].

Peptidoglycan of Staphylococcus aureus cell wall impact on the both estrogen-treated and pseudopragnant rat myometrium contractile activity dynamics was studied. The results of experiments shown that rat myomatrium sensitivity to peptidoglican depends on the hormonal background. Peptidoglycan modified the myometrium contractile activity of both estrogen-treated and pseudopragnant rats. In estrogen-treated rat myometrium peptidoglycan reduces frequency and duration of contractions (elongated the uterine cycle) while in the pseudopregnant rat myometrium it increased the amplitude as well as the duration and the freqwency (deshortened the uterine cycle). During the experiments we found that peptidoglycan has stronger uterotonic effect than prostaglandin F2α. In this connection we conclude that the changes ofmyometral contractile activity after peptidoglycan action may have negative effects for fertility and course of pregnancy.

[Comparative effects of menthol and icilin on the induced contraction of the smooth muscles of the vas deferens of normal and castrated rats].

Non-specific TRPM8 agonist menthol was shown to inhibit voltage- and agonist-evoked contractions of the smooth muscle (SM) of rat vas deferens. Here we compared the action of menthol with the action of more specific TRPM8 agonist icilin on depolarization- (60 mM KCl), carbachol-(CCh) and noradrenalin-(Nor)-evoked contractions of the SM strips from the prostatic and epididymal portions of the vas deferens of normal and castrated (60-137 days) rats. Inhibitory action of menthol (100 microM) and icilin (10 microM) on the amplitude of KCl-, CCh- and Nor-induced contractions of normal as well as castrated rats was similar consisting about 50%, despite castration per se strongly potentiated CCh- and Nor-evoked contractions compared to the control animals. In the epididymal portion of the control animals menthol suppressed KC 1- and CCh-evoked contractions by 46 +/- 5% and 32 +/- 3% and icilin by only 14 +/- 4% and 6 +/- 7%, respectively, whilst after castration both compounds became virtually ineffective. Considering that TRPM8 may localize in the sarcolemma and sarcoplasmic reticulum (SR) membrane and that menthol can also block voltage-gated calcium channels (VGCCs), our data indicate that in the prostatic portion TRPM8 modulates contractility by primarily decreasing the SR Ca2+ stores content, whilst in the epididymal one by both decreasing the SR filling and supporting Ca2+ entry. Drop in the circulation androgens as a result of castration changes the menthol- and icilin-mediated modulation of the rat vas deferens SM contractility via the decrease of the expression of L-type VGCCs and increase of the expression of TRPM8.

[Urothelium-dependent modulation of urinary bladder smooth muscle contractions by menthol].

TRPM8 cold receptor/channel is considered amongst the variety of receptors that support and modulate sensory function of urothelium, although the information regarding this is still quite contradictory. Here we have studied the effects of nonspecific TRPM8 activator menthol on the contractions of the smooth muscle strips of the rat bladder with intact and removed urothelium, and assessed the expression in them of TRPM8 mRNA using semi-quantitative RT-PCR. Menthol (100 microM) decreased the basal tone and the amplitude of spontaneous contractions only in the strips with intact urothelium. Irrespective of the presence of urothelium it similarly inhibited (by approximately 45 %) the contractions evoked by high-potassium depolarization. Contractions induced by muscarinic agonist carbachol (1 microM) were inhibited by menthol much stronger (by approximately 63%) if the urothelium was present than without it (by approximately 12%). Expression of TRPM8 mRNA in urothelium was not detected, whilst in detrusor smooth muscle it was found very low. We conclude that modulation of contractile responses by menthol is most likely explained by its blocking action on voltage-gated calcium channels ofdetrusor smooth muscle cells (SMC) and by menthol-stimulated release from urothelium of some factor(s) with relaxant effects on SMCs. Stimulation of the secretion of these factors from urothelial cells most likely involves menthol-induced, TRPM8-independent mobilization of calcium.

[Expression of the cold receptor TRPM8 in the smooth muscles of the seminal ejaculatory ducts in rats].

By means of real-time RT-PCR and immunofluorscent staining combined with confocal microscopy we show for the first time the expression ofmRNA and protein ofthe cold/menthol-sensitive cationic channel, TRPM8, in the smooth muscle cells (SMC) from the epididimal and prostatic portions of the rat vas deferens. Expression of TRPM8 mRNA correlated with the expression ofmRNA for androgen receptor (AR): orchidectomy of the animals resulted in the enhancement of the expression of both mRNAs, which likely reflects specific for the vas deferens compensatory response to the decreasing levels of circulating androgens. TRPM8 protein in the SMC from both parts of the vas deferens primarily localized outside the plasma membrane (PM), however, in the SMC from prostatic portion we observed higher TRPM8 protein targeting specifically the endoplasmic reticulum and PM, where it can form functional cold/menthol-sensitive cationic channel capable of modulating contractile activity of the smooth muscle.

[Modulation of the smooth muscle contractions of the rat vas deferens by TRPM8 channel agonist menthol].

TRPM8 is nonselective, Ca2- permeable cationic channel, which is activated by innocuous cold and by chemical drugs imitators of cooling, menthol, icilin and cucalyptol. TRPM8 expression was detected in the smooth muscle cells of the rat vas deference with preferential localization of the TRPM8 protein to the membrane of sarcoplasmic reticulum (SR). In the present work we have studied the effects of TRPM8 channel agonist, menthol, on the contractions of the smooth muscle strips of the epididimal and prostatic portions of the rat vas deferens evoked by potassium rich (KCl) Krebs solution and by muscarinic or adrenergic agonists carbachol (CCh) or noradrenalin (Nor). Menthol (0.1-1 mmol/l) per se virtually unaffected the basal tone, but inhibited in a dose-dependent manner KCl-, CCh- and Nor-evoked contractions of both parts of the vas deference by 30-50%. Blockade of the Ca2+ -ATPase of the SR with cyclopiazonic acid (CPA, 10 micromol/l) enhanced inhibitory action of menthol on KCl-induced contractions, but slightly decreased inhibition by menthol of agonist-induced ones. Nonspecific TRPM8 blocker, capsazepine (10 micromol/l), did not eliminate, but augmented inhibitory action of menthol on all types of contractions. Our data propose that menthol inhibits contractions via two mechanisms: partial blockade of Ca2+ entry via the voltage-gated, L-type calcium channels and a decrease of the calcium storage capacity of the SR. The latter mechanism at least in part is mediated by the SR-resident TRPM8 channel, which by activation of menthol leads to the enhancement of passive leak of Ca2+ from the SR and reduction in the amount of the releasable calcium during activation of contractions.

Internet resources integrating many small-molecule databases.

()New data, tools and services recently made available on the web server (http://cactus.nci.nih.gov) of the Computer-Aided Drug Design (CADD) Group, NCI, NIH, developed in the context of chemoinformatics and drug development work, are presented. These tools are designed for searching for structures in very large databases of small molecules. One of them is a web service-the Chemical Structure Look-up Service (CSLS)-for very rapid structure look-up in an aggregated collection of more than 80 databases comprising more than 27 million unique structures at the time of this writing. CSLS contains pointers to the entries in toxicology-related databases, catalogues of commercially available samples, drugs, assay results data sets, and databases in several other categories. CSLS allows the user to find out very rapidly in which one(s) of all these databases a given structure occurs independent of the representation of the input structure, by making use of InChIs as well as new CACTVS hashcode-based identifiers. These latter, calculable, identifiers are designed to take into account tautomerism, different resonance structures drawn for charged species, and presence of additional fragments. They make possible fine-tunable yet rapid compound identification and database overlap analyses in very large compound collections.

[Transfer factor of immune reactivity to diphtheria-tetanus anatoxin modulates the action of neurotransmitters in the intestinal smooth muscle].

Transfer factor (TF) of immune reactivity (10(-5) - 10(-3) mg/ml) to diphtheria-tetanus anatoxin modulates slow waves and spontaneous contractile activity of non-atropinized smooth muscle stripes (SMS) of guinea-pig taenia coli. TF (10(-4) mg/ml) transforms slow waves into stable depolarization and tonic contraction. After SMS atropinization, the substance acts in the same way. In the presence of methylene blue (10(-5) M), a guanylatecyclase blocker, FT induces transitory increase of SMS muscle tone, which is followed by their stable relaxation. ATP and UTP, purinoceptors agonists, evoke substantial hyperpolarization of smooth muscle cells membrane and their relaxation. FT enhances post-inhibitory excitation in SMS. In the presence of acetylcholine (10(-5) M) FT (10(-4) mg/ml) transforms the inhibitory ATP action on tonic contraction into excitative. This substance (10(-5), 10(-4) mg/ml) enhances Ca2+ mobilization from ryanodine-sensitive calcium store, inhibits the release of these cations from IP3-sensitive calcium store of sarcoplasmic reticulum. TF demolishes the inhibitory actions of sodium nitroprusside (nitric oxide donor), and noradrenaline in taenia coli smooth muscles.

[Effects of cell-binding protein A of Staphylococcus aureus on the level of intracellular calcium ions and actomyosin ATP-ase activity in the smooth muscles].

Immune-active substance of Staphylococcus aureus, cell-bound protein A (CBPA), enhances the acetylcholine- or hyperpotassium (K+) Krebs solution-evoked excitation in Taenia coli smooth muscles. CBPA increases caffeine- and carbachole-evoked Ca2+ signals in smooth muscle cells suspension, loaded with indo-1, and also caffeine- and acetylcholine-evoked contraction in smooth muscles slices. Against a background of CBPA-suppressed action of sodium nitroprusside, ATP evokes the membrane depolarization. CBPA in small concentrations potentiates ATPase (Mg2+,Ca2+-; Mg2+- and Mg2+- in the presence of EGTA) activity of actomyosin in the smooth muscles.

[Effects of Staphylococcus aureus cell-bound protein A on adenosine triphosphate and nitric oxide inhibitory actions in smooth muscles].

Cell-bound protein A (CBPA), an immune-active substance of Staphylococcus aureus was ascertained to depolarize membrane of taenia coli smooth muscle (SM) cells, depress ATP inhibiting action (or uridinetriphosphate (UTP)) and sodium nitroprusside (SNP). ATP or UTP-induced membrane hyperpolarization increased during first minutes of CBPA exposure. Bacterial substance enhanced and then inhibited fast component of nicotine-induced relaxation of histamine-activated smooth muscles. This enhancement was inhibited by N(omega)-nitro-Larginine, a NO-syntase blocker. CBPA decreased ATP inhibiting action upon histamine-induced contraction, but enhanced cholinergic SM excitation. All these processes are reversible.

[Molecular mechanisms of receptor-dependent signaling in the cells of the longitudinal and circular intestinal smooth muscles].

The G-protein-dependent intracellular signal cascades of excitation in longitudinal and circular intestinal smooth muscles (SM) are compared and summarized in the present review. The key mechanism of excitation in longitudinal SM is an activation of electro-mechanical coupling, in that G-proteins, phospholipase A2, arachidonic acid, membrane-bound cyclic adenosine diphosphoribose and Ca2+ are involved. We observed the role of arachidonic acid-activated chorine and voltage-dependent Ca2+ -channels (L-type) in Ca2+ mobilization in these muscle cells. In contrast to longitudinal, the main mechanism of agonist-induced excitation in circular SM is connected with activation of key methabotropical processes. The role of Rho-kinase in mechanisms of Ca2+ --sensitization of contractile apparatus in SM is also shown in this review. A comparative analysis of involvement of different links of signal cascades in initial and sustained phases of contraction in longitudinal and circular SM are also reviewed.

[Transfer factor modulates inhibitory action of neurotransmitters on intestinal smooth muscles]. / Faktor perenosu moduliuie hal'mivnu diiu neiromediatoriv na hladen'ki m'iazy kyshechnyka.

It has been shown that Transfer factor (TF) to Staphilococcus aureus antigens blocked ATP-induced component of the inhibitory junction potential in taenia coli smooth muscle from guinea-pig, and converted an inhibitory action of exogenous ATP into an exciting one (instead of the hyperpolarization of the smooth muscle, TF induced its depolarization). TF at 10-6 mg/ml converted the relaxing effects of sodium nitroprusside (nitric oxide donor) into exciting ones in smooth muscle strips. Higher concentrations (10-5-10-3 mg/ml) of TF slightly amplified the relaxing effect of sodium nitroprusside. Both a- and b-adrenergic activation in taenia coli smooth muscle were not sensitive to that agent.

Solution structure of the p53 regulatory domain of the p19Arf tumor suppressor protein.

Arf is a tumor suppressor that regulates p53 function and is a frequent target for loss in human cancers. Through two novel mechanisms, Arf inhibits the oncoprotein Hdm2, a negative regulator of p53. (1) Arf inhibits the E3 ubiquitin ligase activity of Hdm2 that leads to p53 degradation, and (2) Arf sequesters Hdm2 within nucleoli. These activities of Arf promote p53-mediated cell cycle arrest and apoptosis. Fundamental to these processes are interactions between Arf and Hdm2. Here we show that a peptide containing the 37 N-terminal amino acids of mouse Arf (mArfN37) localizes to nucleoli, sequesters Hdm2 within nucleoli, and causes cell cycle arrest. Circular dichroism and NMR spectroscopy show that mArfN37 is largely unstructured under aqueous conditions; however, the peptide adopts two alpha-helices (helix 1, residues 4-14; and helix 2, residues 20-29) in 2,2,2-trifluoroethanol (TFE). Each helix contains an amino acid motif that is repeated twice in mArfN37, once in each helix. The two helices, however, do not interact but are connected by an apparently flexible linker. The repeated motif contains Arg residues spaced by a hydrophobic segment that may be involved in Hdm2 recognition and binding. The RRPR nucleolar localization signal, contained within residues 31-34, appears to be disordered under all conditions. The identification of two Arf structural modules suggests that short peptides containing the repeated motif may function as Arf mimics and may allow the design of small molecule Arf mimics in the future.