The upgraded TOMAS device: A toroidal plasma facility for wall conditioning, plasma production, and plasma-surface interaction studies.

The Toroidal Magnetized System device has been significantly upgraded to enable development of various wall conditioning techniques, including methods based on ion and electron cyclotron (IC/EC) range of frequency plasmas, and to complement plasma-wall interaction research in tokamaks and stellarators. The toroidal magnetic field generated by 16 coils can reach its maximum of 125 mT on the toroidal axis. The EC system is operated at 2.45 GHz with up to 6 kW forward power. The IC system can couple up to 6 kW in the frequency range of 10 MHz-50 MHz. The direct current glow discharge system is based on a graphite anode with a maximum voltage of 1.5 kV and a current of 6 A. A load-lock system with a vertical manipulator allows exposure of material samples. A number of diagnostics have been installed: single- and triple-pin Langmuir probes for radial plasma profiles, a time-of-flight neutral particle analyzer capable of detecting neutrals in the energy range of 10 eV-1000 eV, and a quadrupole mass spectrometer and video systems for plasma imaging. The majority of systems and diagnostics are controlled by the Siemens SIMATIC S7 system, which also provides safety interlocks.

[Genetic characterization of Vibrio cholerae strains emerging in Russian Federation during 7th cholera pandemic].

AIM: Comparative molecular-genetic analysis of clinical Vibrio cholerae eltor biovariant strains isolated in Russia during various years. MATERIALS AND METHODS: Microbiological and biochemical methods were used for studies of 25 clinical strains of classic and eltor biovariant cholera, PCR testing and sequencing of various genes was also performed. RESULTS: Phenotypic and genetic analysis of clinical V. cholerae strains isolated in Russia during 7th cholera pandemic has confirmed that they belong to biovariant eltor. PCR testing of 21 isolates obtained from patients in 1970 - 2010 has shown that epidemic complications in Russia from 1993 were caused by altered V. cholerae biovariant eltor. Presence of classic cholera biovariant ctxB coding gene in cholera toxin coding CTX prophage is the genetic alteration of these variants, ctxB sequencing in altered variants has confirmed PCR data and shown 2 ctxB gene alleles (ctxB1 and ctxB7). Altered variants produced significantly more cholera toxin than typical strains. CONCLUSION: In 1970 - 2010 67.6% of clinical isolates were altered V. cholera biovariant eltor variants. These new variants were genetically diverse. Alteration of cholera eltor biovariant genome caused toxigenicity increase.

[Variation in the genome of CTXphi prophage of Vibrio cholerae biovar El Tor caused by Tn5-Mob transposon].

A key pathogenicity factor of the cholera etiologic agent is cholera toxin (CT) whose synthesis is encoded by the ctxAB operon forming apart of the CTXphi ptophage. Alterations in the virulent properties of the cholera vibrios are based on the variability of the CTXphi prophage containing the genes for ctxAB, zot, ace, cep, orfU, and psh in its core region. At the same time, the mechanism of the porophage genome reorganization needs further and more profound analysis. The goal of this work was to demonstrate that transposon Tn5-Mob (Kmr), when introduced into the chromosome of the V. cholera model strain MAK757 El Tor biovar containing two copies of the CTXphi prophage provoked a reorganization in the CTXphi prophage consisting in the deletion of zot, ace, cep, orfU genes. The level of the CT biosynthesis in the insertion mutants MAK757 chr::Tn5-Mob still retaining only the ctxAB operon, increased more than 2000 times as compared to that of the original strain. The enhanced CT production was shown to be associated with the altered structure of the chromosomal DNA region containing one copy of the ctxAB operon encoding this protein biosynthesis. The mutation in the CTXphi genome induced by Tn5-Mob was unstable. Among 600 isolated colonies obtained after dissemination of the MAK757 chr::Tn5-Mob transposant capable of CT overproduction in the full medium with no antibiotics, 5.8% gave clones that in parallel to the loss of Kmr marker, appeared to be deprived of the ctxAB operon thus becoming non-toxinogenic. The observed formation of the V. cholerae insertion mutants both capable of CT overproduction and non-toxinogenic ones, may be indicative of an important role played in the evolution of the cholera pathogen by the CTXphi genome variability induced by Tn elements. The plasmidless V. cholerae El Tor strain characterized by type II CT hyperproduction thus obtained in our experiments could be used for the production of this protein routinely applied to construct efficient cholera diagnostic and prophylactic preparations.