Pharmacological Analysis of GABA Receptor and Sigma1R Chaperone Interaction: Research Report I-Investigation of the Anxiolytic, Anticonvulsant and Hypnotic Effects of Allosteric GABA Receptors' Ligands.

Two groups of facts have been established in previous drug development studies of the non-benzodiazepine anxiolytic fabomotizole. First, fabomotizole prevents stress-induced decrease in binding ability of the GABA receptor's benzodiazepine site. Second, fabomotizole is a Sigma1R chaperone agonist, and exposure to Sigma1R antagonists blocks its anxiolytic effect.

Involvement of Chaperone Sigma1R in the Anxiolytic Effect of Fabomotizole.

Sigma-1 receptor (chaperone Sigma1R) is an intracellular protein with chaperone functions, which is expressed in various organs, including the brain. Sigma1R participates in the regulation of physiological mechanisms of anxiety (Su, T. P.

The New Dipeptide TSPO Ligands: Design, Synthesis and Structure-Anxiolytic Activity Relationship.

The translocator protein (TSPO, 18 kDa) plays an important role in the synthesis of neurosteroids by promoting the transport of cholesterol from the outer to the inner mitochondrial membrane, which is the rate-limiting step in neurosteroidogenesis. Stimulation of TSPO by appropriate ligands increases the level of neurosteroids. The present study describes the design, synthesis and investigation of anxiolytic-like effects of a series of -acyl-tryptophanyl-containing dipeptides.

Crystal structure and receptor-interacting residues of MYDGF - a protein mediating ischemic tissue repair.

Myeloid-derived growth factor (MYDGF) is a paracrine-acting protein that is produced by bone marrow-derived monocytes and macrophages to protect and repair the heart after myocardial infarction (MI). This effect can be used for the development of protein-based therapies for ischemic tissue repair, also beyond the sole application in heart tissue. Here, we report the X-ray structure of MYDGF and identify its functionally relevant receptor binding epitope.

A Novel Dipeptide Ligand of TSPO.

On the basis of the first dipeptide ligand of TSPO, N-carbobenzoxy-L-tryptophanyl-L-isoleucine amide (GD-23), which was obtained by us earlier, we synthesized a new dipeptide, N-phenylpropionyl-L-tryptophanyl-L-leucine amide (GD-102). GD-102 exhibited anxiolytic activity in the open field test in BALB/c mice and in the elevated plus maze test in ICR mice. The minimum effective dose of GD-102 was one order of magnitude lower than that of GD-23.

Optimizing NBE PK/PD assays using the Gyrolab Affinity Software; conveniently within the bioanalyst's existing workflow.

Aim: The fully automated microfluidics-based Gyrolab is a popular instrument for the bioanalysis of protein therapeutics; requiring minimal sample and reagent volumes. Gyros offers affinity software for determining binding affinity in solution using a high-throughput method and miniaturized reactions.

Results: Using this affinity software, multiple CTGF-targeting reagents were characterized on the Gyrolab after <100% target coverage was seen in a cynomolgus pharmacokinetic/PD study dosed with anti-CTGF antibodies.

Maximizing in vivo target clearance by design of pH-dependent target binding antibodies with altered affinity to FcRn.

Antibodies with pH-dependent binding to both target antigens and neonatal Fc receptor (FcRn) provide an alternative tool to conventional neutralizing antibodies, particularly for therapies where reduction in antigen level is challenging due to high target burden. However, the requirements for optimal binding kinetic framework and extent of pH dependence for these antibodies to maximize target clearance from circulation are not well understood. We have identified a series of naturally-occurring high affinity antibodies with pH-dependent target binding properties.

[The adaptive changes of homeostasis under the influence of high-intensity physical exercises].

The present article was designed to report the results of the investigation into various non-specific adaptive reactions of the organism (NARO) based on the determination of the ratio of lymphocyte to neutrophil levels in the blood of the athletes under conditions of high-intensity training. We evaluated the frequency of different types of adaptive responses in the biathlon skiers at different stages of their annual training cycle. The study has demonstrated the relationship between metabolic changes under the influence of training and the type of adaptive responses.

A high-throughput scintillation proximity assay for sphingosine-1-phosphate lyase.

The emergence of sphingosine-1-phosphate lyase (SPL) as a promising therapeutic target for inflammatory diseases has heightened interest in the identification of small molecules that modulate its activity. The enzymatic activity of SPL is typically measured using radiometric or fluorescence-based assays that require a lipid extraction step, or by direct quantitation of reaction products using mass spectrometry (MS). To facilitate testing large numbers of compounds to identify SPL modulators, we developed a robust scintillation proximity assay (SPA) that is compatible with high-throughput screening (HTS).

The oxidase activity of vascular adhesion protein-1 (VAP-1) is essential for function.

Vascular adhesion protein-1 (VAP-1) has been implicated in the pathogenesis of inflammatory diseases and is suggested to play a role in immune cell trafficking. It is not clear whether this effect is mediated by the oxidase activity or by other features of the protein such as direct adhesion. In order to study the role of VAP-1 oxidase activity in vivo, we have generated mice carrying an oxidase activity-null VAP-1 protein.

[Experience of Lucentis use in patients with wet age-related macular degeneration].

This article provides information about development and introduction of regional standard of specialized medical care for patients with neovascular age-related macular degeneration in medical practice in Tumen region. It discovered new opportunities for improvement of ophthalmologic care in the region.

Bioluminescent method for assaying multiple semicarbazide-sensitive amine oxidase (SSAO) family members in both 96- and 384-well formats.

Vascular adhesion protein-1 (VAP-1), also known as semicarbazide-sensitive amine oxidase (SSAO) or copper-containing amine oxidase (AOC3, EC 1.4.3.

Kinetic characterization and in vitro toxicity evaluation of a luciferase less susceptible to HPV chemical inhibition.

Enzyme-based in vitro toxicity assays are often susceptible to inhibition by test compounds. A mutant luciferase selected to be less susceptible to inhibition by chloroform (CNBLuc03-06) and other high production volume (HPV) chemicals, consisting of three point mutations was created and characterized. The mutant luciferase was less inhibited by chloroform, other HPV chemicals and common surfactant release reagents (Triton-X and SDS) compared to the wild-type.

Creating a mutant luciferase resistant to HPV chemical inhibition by random mutagenesis and colony-level screening.

Firefly luciferase covers a wide range of applications. One common usage of the bioluminescence assay is the measurement of intracellular concentration of adenosine triphosphate (ATP) for cell viability. However, inhibition of the enzyme reaction by chemicals in the assay has so far limited the application of luciferase for high production volume (HPV) chemical testing.

[A chamber for evaluating the free choice by laboratory rats and mice of a space containing tobacco smoke].

An original experimental setup has been designed that allows evaluation of the free preference of space containing tobacco smoke by small laboratory animals. For the laboratory mice (C57B1/6 and BALB/c) and rats (MNRA and MR) placed every day into this box, no emotional-stress reaction (ESR) caused by the environment novelty was observed on the 19th day of experiment. A difference in the free preference of space containing tobacco smoke was observed between inbred animals with active and passive ESR phenotypes.

Alteration of human leukotriene A4 hydrolase activity after site-directed mutagenesis: serine-415 is a regulatory residue.

Leukotriene A4 hydrolase (LTA-H) is a bifunctional protein that has aminopeptidase activity, but also contains an epoxide hydrolase activity that converts leukotriene (LT)A4 to LTB4. The lipid metabolic activity of this enzyme plays a central role in the control of polymorphonuclear leukocyte function and in the development of inflammation. LTA-H is widely spread in many mammalian tissues, although it appears to be inactive in many cases.

Isoprenaline can activate the acetylcholine-induced K+ current in canine atrial myocytes via Gs-derived betagamma subunits.

1. G protein betagamma subunits activate the acetylcholine-induced potassium current IK,ACh. There is no evidence of specificity at the level of the betagamma subunits.

Regulation of leukotriene A4 hydrolase activity in endothelial cells by phosphorylation.

Endothelial cells contain leukotriene (LT) A4 hydrolase (LTA-H) as detected by Northern and Western blotting, but several studies have been unable to detect the activity of this enzyme. Since LTA-H could play a key role in determining what biologically active lipids are generated by activated endothelium during the inflammatory process, we studied possible mechanisms by which this enzyme may be regulated. We find that LTA-H is phosphorylated under basal conditions in human endothelial cells and in this state does not exhibit epoxide hydrolase activity (i.

[The determination of the oxidation phenotype in inbred C57Bl/6 and BALB/c mice].

Antipyrine oxidation was studied in C57BL/6 and BALB/c inbred mice. It was found that C57BL/6 are weak oxidant but BALB/c are strong oxidants of antipyrine. Animals F1 hybrids inherited the high capacity of antipyrine oxidation.

[Neuropsychological studies in the surgery of epilepsy].

Comparison of the findings of neuropsychological examination with the EEG pattern, the results of psychopathological examination, and the character of the paroxysms showed the high informativeness of standardized neuropsychological diagnostic methods in determining the location of the dominant epileptic focus and predicting the outcomes of surgical treatment of patients with epilepsy as well as the transient character of possible disorders of high cortical functions caused by surgical interventions.

[Properties of creatine kinase from skeletal muscle mitochondria].

Creatine kinase from pigeon breast muscle was obtained in a homogeneous (as evidenced from polyacrylamide gel SDS electrophoresis) state. The molecular mass of the enzyme monomer is 43,000. Ultracentrifugation in a sucrose density gradient and gel filtration revealed that the enzyme is present in solution as a mixture of two major forms, i.

[Radiochromatographic study of the distribution of sidnocarb-14C and its metabolites in inbred mice].

The title study permitted detecting genetic differences in sydnocarb metabolism. It was established that the rate of sydnocarb oxidation in BALB/c mice was higher than in C57B1/6 mice.

Studies of the interaction of mitochondrial creatine kinase with the mitochondrial membrane.

In this study we have investigated the binding characteristics of creatine kinase (CK) with mitochondrial membrane. Creatine kinase was found to bind membranes by electrostatic forces. At physiological pH, the interaction seems to be between positively charged groups on the enzyme protein and negatively charged groups on the membrane.