Optogenetic cytosol acidification of mammalian cells using an inward proton-pumping rhodopsin.

Ion gradients are a universal form of energy, information storage and conversion in living cells. Advances in optogenetics inspire the development of novel tools towards control of different cellular processes with light. Rhodopsins are perspective tools for optogenetic manipulation of ion gradients in cells and subcellular compartments, controlling pH of the cytosol and intracellular organelles. The key step of the development of new optogenetic tools is evaluation of their efficiency. Here, we used a high-throughput quantitative method for comparing efficiency of proton-pumping rhodopsins in Escherichia coli cells. This approach allowed us to show that an inward proton pump xenorhodopsin from Nanosalina sp. (NsXeR) is a powerful tool for optogenetic control of pH of mammalian subcellular compartments. Further, we demonstrate that NsXeR can be used for fast optogenetic acidification of the cytosol of mammalian cells. This is the first evidence of optogenetic cytosol acidification by an inward proton pump at physiological pH values. Our approach offers unique opportunities to study cellular metabolism at normal and pathological conditions and might help to understand the role of pH dysregulation in cellular dysfunctions.

Effect of Synthetic Organoselenium Drug on the Degree of Pathological Changes in the Organs of White Mice Immunized with Tularemia and Brucellosis Vaccines.

We studied the effect of the organoselenium compound 2,6-dipyridinium-9-selenium-bicyclo[ 3,3,1]nonan dibromide (974zh) on the severity of pathological changes in the organs of experimental animals immunized with live tularemia and brucellosis vaccines. It was found that 974zh reduced reactogenicity of vaccines for experimental animals. Our findings indicate the prospects for further studies of the effects of 974zh on the functional state of experimental animals.

Immunogenicity of Different Forms of Middle East Respiratory Syndrome S Glycoprotein.

The Middle East respiratory syndrome coronavirus (MERS-CoV) was identified in 2012 during the first Middle East respiratory syndrome (MERS) outbreaks. MERS-CoV causes an acute lower-respiratory infection in humans, with a fatality rate of ~35.5%. Currently, there are no registered vaccines or means of therapeutic protection against MERS in the world. The MERS-CoV S glycoprotein plays the most important role in the viral life cycle (virus internalization). The S protein is an immunodominant antigen and the main target for neutralizing antibodies. In the present study, the immunogenicities of five different forms of the MERS-CoV S glycoprotein were compared: the full-length S glycoprotein, the full-length S glycoprotein with the transmembrane domain of the G glycoprotein of VSV (S-G), the receptor-binding domain (RBD) of the S glycoprotein, the membrane-fused RBD (the RBD fused with the transmembrane domain of the VSV G glycoprotein (RBD-G)), and the RBD fused with Fc of human IgG1 (RBD-Fc). Recombinant vectors based on human adenoviruses type 5 (rAd5) were used as delivery vehicles. Vaccination with all of the developed rAd5 vectors elicited a balanced Th1/Th2 response in mice. The most robust humoral immune response was induced after the animal had been vaccinated with the membrane-fused RBD (rAd5-RBD-G). Only immunization with membrane forms of the glycoprotein (rAd5-S, rAd5-S-G, and rAd5-RBD-G) elicited neutralizing antibodies among all vaccinated animals. The most significant cellular immune response was induced after vaccination of the animals with the full-length S (rAd5-S). These investigations suggest that the full-length S and the membrane form of the RBD (RBD-G) are the most promising vaccine candidates among all the studied forms of S glycoprotein.

Features of emotional and social behavioral phenotypes of calsyntenin2 knockout mice.

Calsyntenin-2 (Clstn2) is the synaptic protein that belongs to the super family of cadherins, playing an important role in learning and memory. We recently reported that Clstn2 knockout mice (Clstn2-KO) have a deficit of GABAergic interneurons coupled with hyperactivity and deficient spatial memory. Given, that impaired functioning of GABA receptors is linked to several psychopathologies, including anxiety and autism, we sought to further characterize Clstn2-KO mice with respect to emotional and social behavior. Clstn2-KO males and females were tested in the elevated plus-maze (EPM), open field (OF), forced swim test, social affiliation and recognition test, social transmission of food preference (STFP), dyadic social interactions and marble burying test. Clstn2-KO mice demonstrated high exploration and hyperactivity in the dimly lit EPM that affect anxiety parameters. In contrast, in a more adverse situation in the OF have increased emotionality in Clstn2-KO males, not females. Assessment of hyperactivity for prolong period in the OF showed that Clstn2-KO animals were able to decline their hyperactivity, but their ambulation still remained higher than in WT littermates. Additionally, Clstn2-KO mice expressed stereotyped behavior. Strikingly, analysis of social behavior identified deficient social motivation and social recognition only in Clstn2-KO males, but not in females. Further analysis of social communication in the STFP and direct observation of agonistic interactions confirmed the reduced social behavior in Clstn2-KO males. Altogether, current results showed Clstn2 gene and sex interactions on socio-emotional performance in mice, suggesting a possible role of calsyntenin2 in psychopathological mechanisms of autism.

[Can the local energy minimization refine the PDB structures of different resolution universally?].

The local energy minimization was statistically validated as the refinement strategy for PDB structure pairs of different resolution. Thirteen pairs of structures with the only difference in resolution were extracted from PDB, and the structures of 11 identical proteins obtained by different X-ray diffraction techniques were represented. The distribution of RMSD value was calculated for these pairs before and after the local energy minimization of each structure. The MMFF94 field was used for energy calculations, and the quasi-Newton method was used for local energy minimization. By comparison of these two RMSD distributions, the local energy minimization was proved to statistically increase the structural differences in pairs so that it cannot be used for refinement purposes. To explore the prospects of complex refinement strategies based on energy minimization, randomized structures were obtained by moving the initial PDB structures as far as the minimized structures had been moved in a multidimensional space of atomic coordinates. For these randomized structures, the RMSD distribution was calculated and compared with that for minimized structures. The significant differences in their mean values proved the energy surface of the protein to have only few minima near the conformations of different resolution obtained by X-ray diffraction for PDB. Some other results obtained by exploring the energy surface near these conformations are also presented. These results are expected to be very useful for the development of new protein refinement strategies based on energy minimization.

Acoustic aspects of the formation of speech in children in the third year of life.

This study covers the third year of life as part of a longitudinal investigation of the establishment of speech in Russian children performed on the basis of listener, phonetic, and instrumented acoustical analysis. The present report addresses the establishment of those additional acoustic and phonetic characteristics in children's speech which allow speech recognition. This is the first instrumented analysis in Russian children with statistical assessment of the dynamics of vowel formants in children's words, of the establishment of characteristics (stress, lack of stress), opposition (palatalization, lack of palatalization of consonants), and voice onset time for plosive consonants. The results showed that recognition of children's words by listeners with a high probability of success resulted from the formation of a system of acoustically stable properties in the children's speech which together provide informational adequacy for verbal communication.

[Diagnostics and treatment of insomnia with concomitant arterial hypertension. Use of somnol as an adjuvant remedy]. / Diagnostika i lechenie insomnii pri arterial'noi gipertenzii. Primenenie Somnola kak ad'iuvantnogo sredstva.

Based upon the conception of "circular-type addiction" scientists mark out psychological (alexithimia), psychopathological (anxiety and depressive disorders), neurological (insomnia) and somatic (circulator disturbances) levels in pathogenesis of arterial hypertension. Sleep disorder have been noted to be a key symptom in phenomenology of arterial hypertension therefore it is recommended giving Somnol as an adjuvant remedy for treatment of arterial hypertension.

[The acoustic aspect of the speech development in children during the third year of life]. / Akusticheskii aspekt formirovaniia rechi rebenka na tret'em godu zhizni.

The current part of a Russian language acquisition longitudinal study based on auditory, phonetic and instrumental analysis is devoted to the third year of child's life. We examined the development of supplementary acoustic and phonetic features of the child's speech providing for the possibility for the speech to be recognized. The instrumental analysis and statistical processing of vowel formant dynamics as well as stress, palatalization and VOT development, has been performed for the first time in Russian children. We showed that the high probability of children words recognition by auditors was due to establishment of a system of acoustically stable features which, in combination with each other, provide for the informative sufficiency of a message.

[Diagnostics and treatment of sleep disorders in elderly people]. / Diagnostika i lechenie narusheniia sna u lits pozhilogo voztasta.

A meta-analysis was done of the published literature concerning the modern status of the issue of sleep disturbances and of findings from the author's investigations. Data are submitted obtained in a clinical examination of 61 elderly patient in a therapeutical hospital. In the above patients, high prevalence has been revealed of different sleep disturbances. Insomnia is often a sequela of psychophysiological derangements, which fact necessitates an exhaustive differential diagnosis and choice of remedies to be used in therapy of the above disorders. An experience is described of employment of zopiclon (Somnol Grindex) to deal with sleep problems in those patients in the involutional period. It is suggested that psychiatrist services be more actively used to give the necessary advice to those patients of the cardiological profile presenting with sleep disturbances.

[Combined therapy of experimental tumors with the vaccinal strain of Venezulean encephalomyelitis virus]. / Kombinirovannaia terapiia éksperimental'nykh opukholei vaktsinnym shtammom virusa venesuél'skogo éntsefalomielita.

The potential of therapy with vaccinal strain of Venezuelan encephalomyelitis virus (VEL) in conjunction with cytostatics (cyclophosphamide) or immunomodulators (T-activin) has been studied. It was found that VEL in conjunction with cyclophosphamide inhibited the antitumor action of the drugs while T-activin potentiated the same effects of the virus and its oncolysate.

Thalicosides A1-A3, minor cycloartane bisdesmosides from Thalictrum minus.

Three new cycloartane bisdesmosides, two of which are based on a new genin, were isolated from the above-ground parts of Thalictrum minus. Thalicosides A1-A3 (1-3) were characterized as 3-O-beta-D-galactopyranosyl-29-O-beta-D-glucopyranosyl-3beta,16beta++ +, 29-trihydroxy-22(S),25-epoxycycloartane (1); 3-O-alpha-L-arabinopyranosyl-29-O-beta-D-glucopyranosyl-3beta,1 6beta, 29,22(S)-tetrahydroxycycloart-24-ene (2); and 3-O-alpha-L-arabinopyranosyl-29-O-beta-D-glucopyranosyl-3beta,1 6beta, 29-trihydroxy-22(S),25-epoxycycloartane (3), respectively. The structural assignments of these new compounds were based on interpretation of spectroscopic data. Thalicoside A2 showed in vitro inhibition of the fungus Candida albicans and also activity against Staphylococcus aureus.

Squarroside C, a new cycloartene bisdesmoside from Thalictrum squarrosum.

Squarroside C (1), a new cycloartane 3,21-bisdesmoside, was isolated from the above-ground parts of Thalictrum squarrosum. The structure of 1 was established as 3-O-[O-alpha-L-rhamnopyranosyl-(1-->6)-beta-D-glucopyranosyl]-21-O-be ta-D-glucopyranosyl-21(S),22(S),23(R),3beta,21alpha,22beta, 30-tetrahydroxy-21,23-epoxycycloart-24-ene by 2D NMR spectroscopy and FABMS.

The elucidation of the structure of thalicoside F, A. minor oleanane glycoside from Thalictrum minus L.

A new triterpenoid saponin, thalicoside F was isolated from the above-ground part of Thalictrum minus L. (Ranunculaceae family) collected in Eastern Siberia (Russia). The structure of thalicoside F was elucidated by a combination of 1D- and 2D-NMR spectroscopy and mass spectroscopy, and was determined to be C47H74O17{3-beta-O-[alpha-L-rhamnopyranosyl-(1-->2)-beta-D- glucopyranosyl-(1-->4)-alpha-L-arabinopyranosyl]-11 alpha, 12 alpha-epoxyoleanane-28,13 beta-olide}.