Work on PETS Developed at CIEMAT

CIEMAT has been working on the RF power extractor so-called PETS (Power Extraction and Transfer Structure) for the CLIC Test Facility 3 (CTF3) since 2007. The first contribution has been installed at the Test Beam Line (TBL). Additionally, a new PETS configuration is presently under fabrication at CIEMAT and will be installed in the Test Module at CTF3. This paper describes the PETS prototypes design, fabrication and assembly techniques. The characterization of the devices with low RF power is also described.Comment: 9 pages, 9 figures, 3 tables, 10 references. Work presented in the LCWS1

Influence of Water Factor on the Incidence (XI, XIV, XIV) Classes of Diseases Among Adult Population in the Rural Taxons

Purpose of research: correlation analysis influence of some indicators of drinking water quality (total hardness, dry residue, Cl-, SO4-, salts of Ca, Mg, F, Al, Fe, Zn, Cu, Mn, nitrogen ammonia, nitrites, pH, nitrates, oxidation) to the incidence (XI, XIV, XIV) classes of diseases among adult population in the rural taxons of Dnipropetrovsk region for 2008 - 2013 years. Materials and Methods. Statistical processing and analysis results of research carried out by the medical statistical methods. Evaluation of the relationship between variables was carried out by coefficients of Spearman's rank correlation (r). Research methods: sanitary-toxicological, physical-chemical (for definition indicators of potable water quality from decentralized water supply sources); medico - statistical (mathematical processing obtained quantitative indicators, methods of variation statis­tics). Results of research and Discussion. In the local water sources from some rural taxons was shown, that salt arthropathy among adults was weakly correlated with all inorganic substances, except: pH + F + oxidability (r=0.01-0.28, p < 0.05). A similar trend was observed between salt arthropathy, which was correlated with all inorganic substances, except nitrate + oxidability (r= 0.03, p < 0.05). Weak correlation had been shown between a combined effect in water pH + F + nitrates and incidence of the stones of kidney and ureter diseases at the adult population (r=0.03, p <0.05)

Technical Specification for the CLIC Two-Beam Module

A high-energy (0.5-3 TeV centre-of-mass), highluminosity Compact Linear Collider (CLIC) is being studied at CERN [1]. The CLIC main linacs, 21-km long each, are composed of 2-m long two beam modules. This paper presents their current layout, the main requirements for the different sub-systems (alignment, supporting, stabilization, cooling and vacuum) as well as the status of their integration

Human condensin function is essential for centromeric chromatin assembly and proper sister kinetochore orientation.

Condensins I and II in vertebrates are essential ATP-dependent complexes necessary for chromosome condensation in mitosis. Condensins depletion is known to perturb structure and function of centromeres, however the mechanism of this functional link remains elusive. Depletion of condensin activity is now shown to result in a significant loss of loading of CENP-A, the histone H3 variant found at active centromeres and the proposed epigenetic mark of centromere identity. Absence of condensins and/or CENP-A insufficiency produced a specific kinetochore defect, such that a functional mitotic checkpoint cannot prevent chromosome missegregation resulting from improper attachment of sister kinetochores to spindle microtubules. Spindle microtubule-dependent deformation of both inner kinetochores and the HEC1/Ndc80 microtubule-capturing module, then results in kinetochore separation from the Aurora B pool and ensuing reduced kinase activity at centromeres. Moreover, recovery from mitosis-inhibition by monastrol revealed a high incidence of merotelic attachment that was nearly identical with condensin depletion, Aurora B inactivation, or both, indicating that the Aurora B dysfunction is the key defect leading to chromosome missegregation in condensin-depleted cells. Thus, beyond a requirement for global chromosome condensation, condensins play a pivotal role in centromere assembly, proper spatial positioning of microtubule-capturing modules and positioning complexes of the inner centromere versus kinetochore plates

A new assay for measuring chromosome instability (CIN) and identification of drugs that elevate CIN in cancer cells.

BACKGROUND: Aneuploidy is a feature of most cancer cells that is often accompanied by an elevated rate of chromosome mis-segregation termed chromosome instability (CIN). While CIN can act as a driver of cancer genome evolution and tumor progression, recent findings point to the existence of a threshold level beyond which CIN becomes a barrier to tumor growth and therefore can be exploited therapeutically. Drugs known to increase CIN beyond the therapeutic threshold are currently few in number, and the clinical promise of targeting the CIN phenotype warrants new screening efforts. However, none of the existing methods, including the in vitro micronuclei (MNi) assay, developed to quantify CIN, is entirely satisfactory. METHODS: We have developed a new assay for measuring CIN. This quantitative assay for chromosome mis-segregation is based on the use of a non-essential human artificial chromosome (HAC) carrying a constitutively expressed EGFP transgene. Thus, cells that inherit the HAC display green fluorescence, while cells lacking the HAC do not. This allows the measurement of HAC loss rate by routine flow cytometry. RESULTS: Using the HAC-based chromosome loss assay, we have analyzed several well-known anti-mitotic, spindle-targeting compounds, all of which have been reported to induce micronuclei formation and chromosome loss. For each drug, the rate of HAC loss was accurately measured by flow cytometry as a proportion of non-fluorescent cells in the cell population which was verified by FISH analysis. Based on our estimates, despite their similar cytotoxicity, the analyzed drugs affect the rates of HAC mis-segregation during mitotic divisions differently. The highest rate of HAC mis-segregation was observed for the microtubule-stabilizing drugs, taxol and peloruside A. CONCLUSION: Thus, this new and simple assay allows for a quick and efficient screen of hundreds of drugs to identify those affecting chromosome mis-segregation. It also allows ranking of compounds with the same or similar mechanism of action based on their effect on the rate of chromosome loss. The identification of new compounds that increase chromosome mis-segregation rates should expedite the development of new therapeutic strategies to target the CIN phenotype in cancer cells

Contrasting roles of condensin I and condensin II in mitotic chromosome formation

© 2012. Published by The Company of Biologists LtdIn vertebrates, two condensin complexes exist, condensin I and condensin II, which have differing but unresolved roles in organizing mitotic chromosomes. To dissect accurately the role of each complex in mitosis, we have made and studied the first vertebrate conditional knockouts of the genes encoding condensin I subunit CAP-H and condensin II subunit CAP-D3 in chicken DT40 cells. Live-cell imaging reveals highly distinct segregation defects. CAP-D3 (condensin II) knockout results in masses of chromatin-containing anaphase bridges. CAP-H (condensin I)-knockout anaphases have a more subtle defect, with chromatids showing fine chromatin fibres that are associated with failure of cytokinesis and cell death. Super-resolution microscopy reveals that condensin-I-depleted mitotic chromosomes are wider and shorter, with a diffuse chromosome scaffold, whereas condensin-II-depleted chromosomes retain a more defined scaffold, with chromosomes more stretched and seemingly lacking in axial rigidity. We conclude that condensin II is required primarily to provide rigidity by establishing an initial chromosome axis around which condensin I can arrange loops of chromatin.This work was supported by an Australian Research Council discovery project [grant number DP110100784 to D.F.H., K.H.A.C. and W.C.E.]; National Health and Medical Research Council (NHMRC) project grants [APP1030358 and 546454]; an NHMRC RD Wright Fellowship to P.K.; an NHMRC Senior Research Fellowship to C.B.W.; an NHMRC Senior Principal Research Fellowship to K.H.A.C.; and by the Victorian Government’s Operational Infrastructure Support Progra

Human artificial chromosome with a conditional centromere for gene delivery and gene expression.

Human artificial chromosomes (HACs), which carry a fully functional centromere and are maintained as a single-copy episome, are not associated with random mutagenesis and offer greater control over expression of ectopic genes on the HAC. Recently, we generated a HAC with a conditional centromere, which includes the tetracycline operator (tet-O) sequence embedded in the alphoid DNA array. This conditional centromere can be inactivated, loss of the alphoid(tet-O) (tet-O HAC) by expression of tet-repressor fusion proteins. In this report, we describe adaptation of the tet-O HAC vector for gene delivery and gene expression in human cells. A loxP cassette was inserted into the tet-O HAC by homologous recombination in chicken DT40 cells following a microcell-mediated chromosome transfer (MMCT). The tet-O HAC with the loxP cassette was then transferred into Chinese hamster ovary cells, and EGFP transgene was efficiently and accurately incorporated into the tet-O HAC vector. The EGFP transgene was stably expressed in human cells after transfer via MMCT. Because the transgenes inserted on the tet-O HAC can be eliminated from cells by HAC loss due to centromere inactivation, this HAC vector system provides important novel features and has potential applications for gene expression studies and gene therapy

μ-Opioid receptor 6-transmembrane isoform: A potential therapeutic target for new effective opioids

6TM MOR is a functional isoform of μ-opioid receptor.6TM MOR has a different cellular localization with respect to 7TM-MOR.6TM MOR may signal via different cellular pathways with respect to 7TM MOR.6TM MOR stimulation may mediate excitatory cellular effects.The μ-opioid receptor (MOR) is the primary target for opioid analgesics. MOR induces analgesia through the inhibition of second messenger pathways and the modulation of ion channels activity. Nevertheless, cellular excitation has also been demonstrated, and proposed to mediate reduction of therapeutic efficacy and opioid-induced hyperalgesia upon prolonged exposure to opioids. In this mini-perspective, we review the recently identified, functional MOR isoform subclass, which consists of six transmembrane helices (6TM) and may play an important role in MOR signaling. There is evidence that 6TM MOR signals through very different cellular pathways and may mediate excitatory cellular effects rather than the classic inhibitory effects produced by the stimulation of the major (7TM) isoform. Therefore, the development of 6TM and 7TM MOR selective compounds represents a new and exciting opportunity to better understand the mechanisms of action and the pharmacodynamic properties of a new class of opioids

Способы управления параметрами вибрации пассажирских вагонов

[For the English abstract and full text of the article please see the attached PDF-File (English version follows Russian version)].ABSTRACT When the car building plant in Tver began serial production of new generation passenger cars, its specialists had to face the facts of not always justified modification of, it would seem, already comfortable and safe designs. In particular, weakened vibration parameters during the construction of lounge cars, bars, laboratories were found out. The feasibility of reducing the vibration indicators of the lounge car by increasing the bending stiffness of the body and selecting the most rational vertical damper of the central suspension is given. Keywords: railway, car building, lounge car, vibration, smooth running, bending stiffness, design, testing, parameters’ control.Текст аннотации на англ. языке и полный текст статьи на англ. языке находится в прилагаемом файле ПДФ (англ. версия следует после русской версии).Когда вагоностроительный завод в Твери начал серийный выпуск пассажирских вагонов нового поколения, его специалистам пришлось столкнуться с фактами не всегда оправданной модификации, казалось бы, уже и без того более комфортных и безопасных конструкций. В частности, обнаружились ослабления вибрационных параметров при постройке вагонных салонов, баров, лабораторий. Дано обоснование возможности уменьшения показателей вибрации вагона-салона за счёт увеличения изгибной жесткости кузова и подбора наиболее рационального вертикального демпфера центрального подвешивания

Исследование прочности кузовов вагонов электропоезда нового поколения

Basing on computational-experimental methods, theoretical and experimental studies were carried out in Tver on carrying capacity of car bodies of electric trains of the new generation EG2Tv under the influence of various normative stresses. Based on the preliminary evaluation of stress-strain state individual components of the metal structure of bodies were changed. The results of static strength tests showed that the main indicators of strength of the body fully comply with the requirements of standards and thus the rationality of the decisions made is confirmed. Taking into account the positive results of the tests, the bodies were transferred for further tests, namely, to determine the parameters of flexural vibrations.В Твери на основе расчётно--экспериментальных методов проведены теоретические и экспериментальные исследования несущей способности кузовов вагонов электропоезда нового поколения ЭГ2Тв при воздействии различных нормативных нагрузок. На основании предварительной оценки напряжённо-деформированного состояния отдельные узлы металлоконструкции кузовов были изменены. Результаты прочностных статических испытаний показали, что по основным показателям прочности кузова полностью соответствуют требованиям нормативов и тем самым подтверждается рациональность принятых решений. С учётом положительных результатов испытаний кузова были переданы для дальнейших испытаний, а именно для определения параметров изгибных колебаний