WorldWideScience

Sample records for qualified nine-color intracellular

  1. Qualified Census Tracts

    Data.gov (United States)

    Department of Housing and Urban Development — A Qualified Census Tract (QCT) is any census tract (or equivalent geographic area defined by the Census Bureau) in which at least 50% of households have an income...

  2. The Fabrication of Qualified Citizens

    DEFF Research Database (Denmark)

    Andrade-Molina, Melissa

    2017-01-01

    a rhizomatic analytical move, a historization of the present is deployed to map the fabrication of the desired qualified citizen in Chile. The analysis evidences the (re)production of dominant narratives about the “qualified citizen” are and have been entangled with the functioning of school geometry...

  3. Qualifying for Career Planning

    Directory of Open Access Journals (Sweden)

    Aleksandra Kovač

    1996-12-01

    Full Text Available The need for education in various spheres of life, which allows a person to fully develop his/her personality, is more and more presen t also in our society. The discussed educational programme Discovering Vocational Goals and Forming Strategies· to Realize them qualifies people for autonomous career planning by offer­ ing them one of the possible ways of making decisions in relation to profession and job. In the course of the realization of the programme the participants are given the chance to think about the kind of work that would suit their interests, capabilities and individual characteristics as weel as the needs in certain environments (possible employers. They are encouraged to make an actual plan, stating how and when the desired objective will be reached. In 1995 the pro­ gramme was being carried out within nine semi­ nars organized at the Employment Offices in Maribor and Ptuj. During that time the pro­ gramme has also been evaluated by Doba - Education Office, which was funded by the Ministry of Work, Family and Social Matters. The paper includes the content of the programme as well as the assessments of the research project. The presented data show that the participants learned the method of career planning and acqu­ired greater self-confidence and motivation to solve their job problem actively.

  4. Qualified Health Plan (QHP) Landscape

    Data.gov (United States)

    U.S. Department of Health & Human Services — QHP Landscape Files present basic information about certified Qualified Health Plans and Stand-alone Dental Plans for individuals-families and small businesses...

  5. Partners in qualified equipment supply

    International Nuclear Information System (INIS)

    Rygg, D.E.; O'Hare, G.J.

    1993-01-01

    Industry initiatives have been taken to improve procurement practices and commercial dedication programs, formation of procurement engineering groups, emphasis on product quality, and increased engineering involvement in procurement and maintenance of qualified equipment. This poses new challenges for many licensees in terms of resources, product knowledge, and access to information normally held proprietary by equipment suppliers. Alternative approaches to future licensee/Westinghouse relationships which will allow licensees to adapt to the changing environment are discussed. 2 figs

  6. 7 CFR 4290.110 - Qualified management.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 15 2010-01-01 2010-01-01 false Qualified management. 4290.110 Section 4290.110... Qualifications for the RBIC Program Organizing A Rbic § 4290.110 Qualified management. An Applicant must show, to the satisfaction of the Secretary, that its current or proposed management team is qualified and has...

  7. ISS qualified thermal carrier equipment

    Science.gov (United States)

    Deuser, Mark S.; Vellinger, John C.; Jennings, Wm. M.

    2000-01-01

    Biotechnology is undergoing a period of rapid and sustained growth, a trend which is expected to continue as the general population ages and as new medical treatments and products are conceived. As pharmaceutical and biomedical companies continue to search for improved methods of production and, for answers to basic research questions, they will seek out new avenues of research. Space processing on the International Space Station (ISS) offers such an opportunity! Space is rapidly becoming an industrial laboratory for biotechnology research and processing. Space bioprocessing offers exciting possibilities for developing new pharmaceuticals and medical treatments, which can be used to benefit mankind on Earth. It also represents a new economic frontier for the private sector. For over eight years, the thermal carrier development team at SHOT has been working with government and commercial sector scientists who are conducting microgravity experiments that require thermal control. SHOT realized several years ago that the hardware currently being used for microgravity thermal control was becoming obsolete. It is likely that the government, academic, and industrial bioscience community members could utilize SHOT's hardware as a replacement to their current microgravity thermal carrier equipment. Moreover, SHOT is aware of several international scientists interested in utilizing our space qualified thermal carrier. SHOT's economic financing concept could be extremely beneficial to the international participant, while providing a source of geographic return for their particular region. Beginning in 2000, flight qualified thermal carriers are expected to be available to both the private and government sectors. .

  8. Qualified Empathy - Project Report 2015-16

    DEFF Research Database (Denmark)

    Austring, Bennye Düranc; Sørensen, Merete Cornet

    2016-01-01

    UCSJ-delen af den afsluttende rapport fra Nordplus-projektet "Qualified Empathy", der blev gennemført sammen med Metropolia, Helsinki, og NTNU, Trondheim, i 2015 og 15.......UCSJ-delen af den afsluttende rapport fra Nordplus-projektet "Qualified Empathy", der blev gennemført sammen med Metropolia, Helsinki, og NTNU, Trondheim, i 2015 og 15....

  9. Nominal Group as Qualifier to "Someone"

    Science.gov (United States)

    Sujatna, Eva Tuckyta Sari; Wahyuni, Sri

    2017-01-01

    The paper titled "Nominal Group as Qualifier to 'Someone'" investigated types of qualifiers which are embedded to the head "someone" in a nominal group. This research was conducted in the light of Systemic Functional Linguistics analysis. The data was analyzed, classified then described using descriptive qualitative method.…

  10. Highly qualified in the Czech Republic

    Directory of Open Access Journals (Sweden)

    Renata Čuhlová

    2017-05-01

    Full Text Available The paper aims to analyse the current situation of the highly qualified in the Czech Republic applying the ISCO qualification. Previous studies suggest that educational attainment has an important impact on labour market performance and national competitiveness. Data analyses approved that highly qualified workforce positively influences economic situation of the EU–15 Member States they work in. However, this hypothesis was disapproved for the EU–10+3 Member States joining the EU after 2004. The difference can be explained by the various stage in convergence process since some New Member States are still in transformation period. We positively appraise the significant growth of fraction of the highly qualified in the Czech Republic since 2004, especially in the major group Professionals involving the most qualified workforce. The same trend happened also for the highly qualified foreigners in the Czech Republic, however their attracting still remains a challenge.

  11. Confidence in leadership among the newly qualified.

    Science.gov (United States)

    Bayliss-Pratt, Lisa; Morley, Mary; Bagley, Liz; Alderson, Steven

    2013-10-23

    The Francis report highlighted the importance of strong leadership from health professionals but it is unclear how prepared those who are newly qualified feel to take on a leadership role. We aimed to assess the confidence of newly qualified health professionals working in the West Midlands in the different competencies of the NHS Leadership Framework. Most respondents felt confident in their abilities to demonstrate personal qualities and work with others, but less so at managing or improving services or setting direction.

  12. Stochastic models of intracellular transport

    KAUST Repository

    Bressloff, Paul C.; Newby, Jay M.

    2013-01-01

    mechanisms for intracellular transport: passive diffusion and motor-driven active transport. Diffusive transport can be formulated in terms of the motion of an overdamped Brownian particle. On the other hand, active transport requires chemical energy, usually

  13. Supply Inventory Management: Evaluation of the Defense Supply Center Columbus Qualified Products List and Qualified Manufacturers List Program

    National Research Council Canada - National Science Library

    2002-01-01

    ... was transitioned from the Military Departments. The Defense Supply Center Columbus currently has management responsibility for over 300 Qualified Products Lists and 4 Qualified Manufacturers Lists...

  14. 26 CFR 1.936-10 - Qualified investments.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 10 2010-04-01 2010-04-01 false Qualified investments. 1.936-10 Section 1.936... (CONTINUED) INCOME TAXES Possessions of the United States § 1.936-10 Qualified investments. (a) In general. [Reserved] (b) Qualified investments in Puerto Rico. [Reserved] (c) Qualified investment in certain...

  15. Insecticide resistance and intracellular proteases.

    Science.gov (United States)

    Wilkins, Richard M

    2017-12-01

    Pesticide resistance is an example of evolution in action with mechanisms of resistance arising from mutations or increased expression of intrinsic genes. Intracellular proteases have a key role in maintaining healthy cells and in responding to stressors such as pesticides. Insecticide-resistant insects have constitutively elevated intracellular protease activity compared to corresponding susceptible strains. This increase was shown for some cases originally through biochemical enzyme studies and subsequently putatively by transcriptomics and proteomics methods. Upregulation and expression of proteases have been characterised in resistant strains of some insect species, including mosquitoes. This increase in proteolysis results in more degradation products (amino acids) of intracellular proteins. These may be utilised in the resistant strain to better protect the cell from stress. There are changes in insect intracellular proteases shortly after insecticide exposure, suggesting a role in stress response. The use of protease and proteasome inhibitors or peptide mimetics as synergists with improved application techniques and through protease gene knockdown using RNA interference (possibly expressed in crop plants) may be potential pest management strategies, in situations where elevated intracellular proteases are relevant. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  16. Solar array qualification through qualified analysis

    Science.gov (United States)

    Zijdemans, J.; Cruijssen, H. J.; Wijker, J. J.

    1991-04-01

    To achieve qualification is in general a very expensive exercise. For solar arrays this is done by a dedicated test program through which final qualification is achieved. Due to severe competition on the solar array market, cheaper means are looked for to achieve a qualified product for the customers. One of the methods is to drastically limit the environmental test program and to qualify the solar-array structure against its environmental loads by analysis. Qualification by analysis is possible. The benefits are that a significant amount of development effort can be saved in case such a powerful tool is available. Extensive testing can be avoided thus saving time and money.

  17. 17 CFR 240.3b-8 - Definitions of “Qualified OTC Market Maker, Qualified Third Market Maker” and “Qualified Block...

    Science.gov (United States)

    2010-04-01

    ... Market Maker, Qualified Third Market Makerâ and âQualified Block Positionerâ. 240.3b-8 Section 240.3b-8... “Qualified Block Positioner”. For the purposes of Regulation U under the Act (12 CFR part 221): (a) The term... inventory turnover in such security. (c) The term Qualified Block Positioner means a dealer who (1) is a...

  18. 46 CFR 390.9 - Qualified withdrawals.

    Science.gov (United States)

    2010-10-01

    ..., construction or reconstruction its fair market value. The party must obtain the prior written permission of the... appraisal to be the fair market value of the vessel, at the time of the acquisition, or the actual cost...: (i) The acquisition, construction or reconstruction of a qualified agreement vessel; (ii) The...

  19. Profitability of Qualified-Labour-Power Production

    Science.gov (United States)

    Baldino, Roberto Ribeiro; Cabral, Tânia Cristina Baptista

    2015-01-01

    In Baldino and Cabral (2013) we introduced the concept of qualified labour-power as the commodity produced by the school system. In the present article we outline a quantitative model to evaluate the profit rate of educational programmes. We compare a medical school programme with a teacher education programme at a public university in Brazil,…

  20. 13 CFR 108.110 - Qualified management.

    Science.gov (United States)

    2010-01-01

    ... 13 Business Credit and Assistance 1 2010-01-01 2010-01-01 false Qualified management. 108.110... management. An Applicant must show, to the satisfaction of SBA, that its current or proposed management team... or proposed management team has sufficient qualifications, SBA will consider information provided by...

  1. The UK system of recognising qualified experts

    International Nuclear Information System (INIS)

    Bines, W.

    2002-01-01

    EURATOM Basic Safety Standards (BSS) Directives have long included requirements for the involvement of qualified experts, the definition of which has scarcely changed since at least 1976. The Directive requirement, in the definition of qualified expert,, for competent authorities to recognise the capacity to act as a qualified expert has been interpreted by Member States in widely differing ways, ranging from the minimalist or case by case to the highly detailed and prescriptive. In the United Kingdom (UK), the qualified expert for occupational radiation protection is the radiation protection adviser and the competent authority is the Health and Safety Executive (HSE). The Ionising Radiations Regulations 1985, which largely implemented the 1980 BSS Directive, required an employer to appoint one or more radiation protection advisers for the purpose of advising him as to the observance of these Regulations and other health and safety matters in connection with ionising radiation. The Regulations addressed the question of recognition by forbidding an employer to appoint a person as a radiation protection adviser unless: that person was suitably qualified and experienced; the employer had notified the Health and Safety Executive in writing of the intended appointment at least 28 days in advance, giving the name of the person and particulars of his qualifications and experience and the scope of the advice he would be required to give; and the employer had received from HSE an acknowledgement in writing of the notification. This system allowed HSE to follow up and query any apparently unsuitable potential appointments while applying a light overall administrative touch. The Approved Code of Practice supporting the Regulations included advice on the qualifications, experience and qualities that the employer should look for in a suitable radiation protection adviser

  2. 33 CFR 154.1026 - Qualified individual and alternate qualified individual.

    Science.gov (United States)

    2010-07-01

    ... alternate must: (1) Be located in the United States; (2) Speak fluent English; (3) Be familiar with the... facility may designate an organization to fulfill the role of the qualified individual and the alternate...

  3. 33 CFR 155.1026 - Qualified individual and alternate qualified individual.

    Science.gov (United States)

    2010-07-01

    ... familiar with the implementation of the vessel response plan; and (4) Be trained in the responsibilities of... vessel may designate an organization to fulfill the role of the qualified individual and alternate...

  4. Reflections on the newly qualified social worker's journey : From university training to qualified practice

    OpenAIRE

    Walker, Clare

    2014-01-01

    This qualitative research study explores the experience of graduating social workers making the transition from university training into work as qualified social work practitioners. Most studies in this area look at the practice readiness of the newly qualified professional. This study looks at the participants’ experience in the work place. How do participants experience this journey of transition? What skills, particularly reflective practice and supervision, learned in training, are import...

  5. The practical skills of newly qualified nurses.

    Science.gov (United States)

    Danbjørg, Dorthe Boe; Birkelund, Regner

    2011-02-01

    This paper reports the findings from a study of newly qualified nurses and which subjects the nurses regarded as the most important in order to be able to live up to the requirements of clinical practice, and how they experience their potential for developing practical and moral skills, after the decrease in practical training. A qualitative approach guided the research process and the analysis of the data. The data was collected by participant observation and qualitative interviews with four nurses as informants. The conclusions made in this study are based on the statements and the observations of the newly qualified nurses. Our findings are discussed in relation to the Aristotelian concept and other relevant literature. The main message is that the newly qualified nurses did not feel equipped when they finished their training. This could be interpreted as a direct consequence of the decrease in practical training. Our study also underlines that the way nursing theory is perceived and taught is problematic. The interviews revealed that the nurses think that nursing theories should be applied directly in practice. This misunderstanding is probably also applicable to the teachers of the theories. Copyright © 2010 Elsevier Ltd. All rights reserved.

  6. Stochastic models of intracellular transport

    KAUST Repository

    Bressloff, Paul C.

    2013-01-09

    The interior of a living cell is a crowded, heterogenuous, fluctuating environment. Hence, a major challenge in modeling intracellular transport is to analyze stochastic processes within complex environments. Broadly speaking, there are two basic mechanisms for intracellular transport: passive diffusion and motor-driven active transport. Diffusive transport can be formulated in terms of the motion of an overdamped Brownian particle. On the other hand, active transport requires chemical energy, usually in the form of adenosine triphosphate hydrolysis, and can be direction specific, allowing biomolecules to be transported long distances; this is particularly important in neurons due to their complex geometry. In this review a wide range of analytical methods and models of intracellular transport is presented. In the case of diffusive transport, narrow escape problems, diffusion to a small target, confined and single-file diffusion, homogenization theory, and fractional diffusion are considered. In the case of active transport, Brownian ratchets, random walk models, exclusion processes, random intermittent search processes, quasi-steady-state reduction methods, and mean-field approximations are considered. Applications include receptor trafficking, axonal transport, membrane diffusion, nuclear transport, protein-DNA interactions, virus trafficking, and the self-organization of subcellular structures. © 2013 American Physical Society.

  7. Characterization of Leptin Intracellular Trafficking

    Directory of Open Access Journals (Sweden)

    E Walum

    2009-12-01

    Full Text Available Leptin is produced by adipose tissue, and its concentration in plasma is related to the amount of fat in the body. The leptin receptor (OBR is a member of the class I cytokine receptor family and several different isoforms, produced by alternative mRNA splicing are found in many tissues, including the hypothalamus. The two predominant isoforms includes a long form (OBRl with an intracellular domain of 303 amino acids and a shorter form (OBRs with an intracellular domain of 34 amino acids. Since OBRl is mainly expressed in the hypotalamus, it has been suggested to be the main signalling form. The peripheral production of leptin by adipocyte tissue and its effects as a signal of satiety in the central nervous system imply that leptin gains access to regions of the brain regulating in energy balance by crossing the blood-brain barrier. In an attempt to characterize the intracellular transport of leptin, we have followed binding internalization and degradation of leptin in HEK293 cells. We have also monitored the intracellular transport pathway of fluorescent conjugated leptin in HEK293 cells. Phenylarsine oxide, a general inhibitor of endocytosis, as well as incubation at mild hypertonic conditions, prevented the uptake of leptin, confirming a receptor-mediated internalization process. When internalized, 125I-leptin was rapidly accumulated inside the cells and reached a maximum after 10 min. After 70 minutes about 40-50% of total counts in each time point were found in the medium as TCA-soluble material. Leptin sorting, at the level of early endosomes, did not seem to involve recycling endosomes, since FITC-leptin was sorted from Cy3- transferrin containing compartments at 37°C. At 45 minutes of continuos internalization, FITC-leptin appeared mainly accumulated in late endocytic structures colocalizing with internalized rhodamine coupled epidermial growth factor (EGF and the lysosomal marker protein lamp-1. The transport of leptin was also shown

  8. 26 CFR 1.132-9 - Qualified transportation fringes.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 2 2010-04-01 2010-04-01 false Qualified transportation fringes. 1.132-9... Qualified transportation fringes. (a) Table of contents. This section contains a list of the questions and answers in § 1.132-9. (1) General rules. Q-1. What is a qualified transportation fringe? Q-2. What is...

  9. 21 CFR 516.141 - Qualified expert panels.

    Science.gov (United States)

    2010-04-01

    ... choose members for the qualified expert panel in accordance with selection criteria listed in paragraph... whether the proposed qualified expert panel meets the selection criteria prior to the panel beginning its... Committee Act, as amended, 5 U.S.C. App. (b) Criteria for the selection of a qualified expert panel. (1) A...

  10. 12 CFR 621.4 - Audit by qualified public accountant.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 6 2010-01-01 2010-01-01 false Audit by qualified public accountant. 621.4... REQUIREMENTS General Rules § 621.4 Audit by qualified public accountant. (a) Each institution shall, at least annually, have its financial statements audited by a qualified public accountant in accordance with...

  11. 11 CFR 9002.11 - Qualified campaign expense.

    Science.gov (United States)

    2010-01-01

    ... 11 Federal Elections 1 2010-01-01 2010-01-01 false Qualified campaign expense. 9002.11 Section 9002.11 Federal Elections FEDERAL ELECTION COMMISSION PRESIDENTIAL ELECTION CAMPAIGN FUND: GENERAL ELECTION FINANCING DEFINITIONS § 9002.11 Qualified campaign expense. (a) Qualified campaign expense means...

  12. 11 CFR 9032.9 - Qualified campaign expense.

    Science.gov (United States)

    2010-01-01

    ... 11 Federal Elections 1 2010-01-01 2010-01-01 false Qualified campaign expense. 9032.9 Section 9032.9 Federal Elections FEDERAL ELECTION COMMISSION PRESIDENTIAL ELECTION CAMPAIGN FUND: PRESIDENTIAL PRIMARY MATCHING FUND DEFINITIONS § 9032.9 Qualified campaign expense. (a) Qualified campaign expense...

  13. 30 CFR 75.155 - Qualified hoisting engineer; qualifications.

    Science.gov (United States)

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Qualified hoisting engineer; qualifications. 75... Persons § 75.155 Qualified hoisting engineer; qualifications. (a)(1) A person is a qualified hoisting engineer within the provisions of subpart O of this part, for the purpose of operating a steam-driven hoist...

  14. 26 CFR 1.30-1 - Definition of qualified electric vehicle and recapture of credit for qualified electric vehicle.

    Science.gov (United States)

    2010-04-01

    ... qualified electric vehicle. A qualified electric vehicle is a motor vehicle that meets the requirements of section 30(c). Accordingly, a qualified electric vehicle does not include any motor vehicle that has ever been used (for either personal or business use) as a non-electric vehicle. (b) Recapture of credit for...

  15. Electron Microscopy of Intracellular Protozoa

    Science.gov (United States)

    1988-12-20

    Classification) " ELECTRON MICROSCOPY OF INTRACELLULAR PROTOZOA 12. PERSONAL AUTHOR(S) Aikawa, Masamichi 13a. TYPE OF REPORT I13b. TIME COVERED 114...authors suggest that anti-CS protein antibody is important in reducing the prevalence of malaria with increasing age among persons in such areas and... Hygine 33, 220-226. 0Giudice, G.D., Engers, H.D., Tougne, C., Biro, S.S., Weiss, N., Verdini, A.S., Pessi, A., Degremont, A.A., Freyvogel, T.A., Lambert

  16. Pathogenic mechanisms of intracellular bacteria.

    Science.gov (United States)

    Niller, Hans Helmut; Masa, Roland; Venkei, Annamária; Mészáros, Sándor; Minarovits, Janos

    2017-06-01

    We wished to overview recent data on a subset of epigenetic changes elicited by intracellular bacteria in human cells. Reprogramming the gene expression pattern of various host cells may facilitate bacterial growth, survival, and spread. DNA-(cytosine C5)-methyltransferases of Mycoplasma hyorhinis targeting cytosine-phosphate-guanine (CpG) dinucleotides and a Mycobacterium tuberculosis methyltransferase targeting non-CpG sites methylated the host cell DNA and altered the pattern of gene expression. Gene silencing by CpG methylation and histone deacetylation, mediated by cellular enzymes, also occurred in M. tuberculosis-infected macrophages. M. tuberculosis elicited cell type-specific epigenetic changes: it caused increased DNA methylation in macrophages, but induced demethylation, deposition of euchromatic histone marks and activation of immune-related genes in dendritic cells. A secreted transposase of Acinetobacter baumannii silenced a cellular gene, whereas Mycobacterium leprae altered the epigenotype, phenotype, and fate of infected Schwann cells. The 'keystone pathogen' oral bacterium Porphyromonas gingivalis induced local DNA methylation and increased the level of histone acetylation in host cells. These epigenetic changes at the biofilm-gingiva interface may contribute to the development of periodontitis. Epigenetic regulators produced by intracellular bacteria alter the epigenotype and gene expression pattern of host cells and play an important role in pathogenesis.

  17. Space-Qualified Traveling-Wave Tube

    Science.gov (United States)

    Wilson, Jeffrey D.; Krawczyk, Richard; Simons, Rainee N.; Williams, Wallace D.; Robbins, Neal R.; Dibb, Daniel R.; Menninger, William L.; Zhai, Xiaoling; Benton, Robert T.

    2010-01-01

    The L-3 Communications Electron Technologies, Inc. Model 999HA traveling-wave tube (TWT), was developed for use as a high-power microwave amplifier for high-rate transmission of data and video signals from deep space to Earth (see figure). The 999HA is a successor to the 999H a non-space qualified TWT described in High-Power, High-Efficiency Ka-Band Traveling-Wave Tube (LEW-17900-1), NASA Tech Briefs, Vol. 31, No. 2 (February 2007), page 32. Operating in the 31.8-to-32.3 GHz frequency band, the 999HA has been shown to generate 252 W of continuous- wave output power at 62 percent overall power efficiency a 75-percent increase in output power over the 999H. The mass of the 999HA is 35 percent less than that of the 999H. Moreover, taking account of the elimination of a Faraday cage that is necessary for operation of the 999H but is obviated by a redesign of high-voltage feed-throughs for the 999HA, the overall reduction in mass becomes 57 percent with an 82 percent reduction in volume. Through a series of rigorous tests, the 999HA has been qualified for operation aboard spacecraft with a lifetime exceeding seven years. Offspring of the 999HA will fly on the Kepler and Lunar Reconnaissance Orbiter missions.

  18. Qualifying cogeneration in Texas and Louisiana

    International Nuclear Information System (INIS)

    Jenkins, S.C.; Cabe, R.; Stauffaeher, J.J.

    1992-01-01

    This paper reports that cogeneration of electricity and useful thermal energy by industrials along the Gulf Coast grew significantly more rapidly than in other parts of the country during and immediately following World War II as a result of the concentration of chemical and plastics processing facilities there. In 1982, Texas passed its version of PURPA, the Public Utility Regulatory Act (PURA) and designated those non-utility generators from which public utilities must purchase electricity as Qualifying Cogenerators. In 1991, there were nearly 7,500 MW of QF power generated for inside-the-fence use or firm capacity sale to utilities, with the two largest utilities in Texas purchasing over half that amount

  19. Qualified market access and inter-disciplinarity

    Directory of Open Access Journals (Sweden)

    Lisa Herzog

    2014-05-01

    Full Text Available This note offers reflections on qualified market access (QMA—the practice of linking trade agreements to values such as human rights, labour standards, or environmental protection. This idea has been suggested by political theorists as a way of fulfilling our duties to the global poor and of making the global economic system more just, and it has influenced a number of concrete policies, such as European Union (EU trade policies. Yet, in order to assess its merits tout court, different perspectives and disciplines need to be brought together, such as international law, economics, political science, and philosophy. It is also worth reflecting on existing practices, such as those of the EU. This note summarises some insights about QMA by drawing such research together and considers the areas in which further research is needed, whilst reflecting also on the merits of interdisciplinary exchanges on such topics.

  20. M2 qualify laser beam propagation

    International Nuclear Information System (INIS)

    Abdelhalim, Bencheikh; Mohamed, Bouafia

    2010-01-01

    One of the most important properties of a laser resonator is the highly collimated or spatially coherent nature of the laser output beam. Laser beam diameter and quality factor M 2 are significant parameters in a wide range of laser applications. This is because the spatial beam quality determines how closely the beam can be focused or how well the beam propagates over long distances without significant dispersion. In the present paper we have used three different methods to qualify the spatial structure of a laser beam propagating in free space, the results are obtained and discussed, and we have found that the Wigner distribution function is a powerful tool which allows a global characterization of any kind of beam

  1. MR imaging of intracellular and extracellular deoxyhemoglobin

    International Nuclear Information System (INIS)

    Janick, P.A.; Grossman, R.I.; Asakura, T.

    1989-01-01

    MR imaging was performed on varying concentrations of intracellular and extracellular deoxyhemoglobin as well as varying proportions of deoxyhemoglobin and oxyhemoglobin in vitro at 1.5T with use of standard spin-echo and gradient-refocused spin sequences. This study indicates that susceptibility-induced T2 shortening occurs over a broad range of intracellular deoxyhemoglobin concentrations (maximal at hematocrits between 20% and 45%), reflecting diffusional effects at the cellular level. T2* gradient-echo imaging enhances the observed hypointensity in images of intracellular deoxyhemoglobin. The characteristic MR appearance of acute hemotomas can be modeled by the behavior of intracellular and extracellular deoxyhemoglobin and oxyhemoglobin

  2. Dynamics of intracellular information decoding

    International Nuclear Information System (INIS)

    Kobayashi, Tetsuya J; Kamimura, Atsushi

    2011-01-01

    A variety of cellular functions are robust even to substantial intrinsic and extrinsic noise in intracellular reactions and the environment that could be strong enough to impair or limit them. In particular, of substantial importance is cellular decision-making in which a cell chooses a fate or behavior on the basis of information conveyed in noisy external signals. For robust decoding, the crucial step is filtering out the noise inevitably added during information transmission. As a minimal and optimal implementation of such an information decoding process, the autocatalytic phosphorylation and autocatalytic dephosphorylation (aPadP) cycle was recently proposed. Here, we analyze the dynamical properties of the aPadP cycle in detail. We describe the dynamical roles of the stationary and short-term responses in determining the efficiency of information decoding and clarify the optimality of the threshold value of the stationary response and its information-theoretical meaning. Furthermore, we investigate the robustness of the aPadP cycle against the receptor inactivation time and intrinsic noise. Finally, we discuss the relationship among information decoding with information-dependent actions, bet-hedging and network modularity

  3. Dynamics of intracellular information decoding.

    Science.gov (United States)

    Kobayashi, Tetsuya J; Kamimura, Atsushi

    2011-10-01

    A variety of cellular functions are robust even to substantial intrinsic and extrinsic noise in intracellular reactions and the environment that could be strong enough to impair or limit them. In particular, of substantial importance is cellular decision-making in which a cell chooses a fate or behavior on the basis of information conveyed in noisy external signals. For robust decoding, the crucial step is filtering out the noise inevitably added during information transmission. As a minimal and optimal implementation of such an information decoding process, the autocatalytic phosphorylation and autocatalytic dephosphorylation (aPadP) cycle was recently proposed. Here, we analyze the dynamical properties of the aPadP cycle in detail. We describe the dynamical roles of the stationary and short-term responses in determining the efficiency of information decoding and clarify the optimality of the threshold value of the stationary response and its information-theoretical meaning. Furthermore, we investigate the robustness of the aPadP cycle against the receptor inactivation time and intrinsic noise. Finally, we discuss the relationship among information decoding with information-dependent actions, bet-hedging and network modularity.

  4. Secretome of obligate intracellular Rickettsia

    Science.gov (United States)

    Gillespie, Joseph J.; Kaur, Simran J.; Rahman, M. Sayeedur; Rennoll-Bankert, Kristen; Sears, Khandra T.; Beier-Sexton, Magda; Azad, Abdu F.

    2014-01-01

    The genus Rickettsia (Alphaproteobacteria, Rickettsiales, Rickettsiaceae) is comprised of obligate intracellular parasites, with virulent species of interest both as causes of emerging infectious diseases and for their potential deployment as bioterrorism agents. Currently, there are no effective commercially available vaccines, with treatment limited primarily to tetracycline antibiotics, although others (e.g. josamycin, ciprofloxacin, chloramphenicol, and azithromycin) are also effective. Much of the recent research geared toward understanding mechanisms underlying rickettsial pathogenicity has centered on characterization of secreted proteins that directly engage eukaryotic cells. Herein, we review all aspects of the Rickettsia secretome, including six secretion systems, 19 characterized secretory proteins, and potential moonlighting proteins identified on surfaces of multiple Rickettsia species. Employing bioinformatics and phylogenomics, we present novel structural and functional insight on each secretion system. Unexpectedly, our investigation revealed that the majority of characterized secretory proteins have not been assigned to their cognate secretion pathways. Furthermore, for most secretion pathways, the requisite signal sequences mediating translocation are poorly understood. As a blueprint for all known routes of protein translocation into host cells, this resource will assist research aimed at uniting characterized secreted proteins with their apposite secretion pathways. Furthermore, our work will help in the identification of novel secreted proteins involved in rickettsial ‘life on the inside’. PMID:25168200

  5. HYPERTHERMIA, INTRACELLULAR FREE CALCIUM AND CALCIUM IONOPHORES

    NARCIS (Netherlands)

    STEGE, GJJ; WIERENGA, PK; KAMPINGA, HH; KONINGS, AWT

    1993-01-01

    It is shown that heat-induced increase of intracellular calcium does not correlate with hyperthermic cell killing. Six different cell lines were investigated; in four (EAT, HeLa S3, L5178Y-R and L5178Y-S) heat treatments killing 90% of the cells did not affect the levels of intracellular free

  6. 49 CFR 604.7 - Qualified human service organizations.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 7 2010-10-01 2010-10-01 false Qualified human service organizations. 604.7... organizations. (a) A recipient may provide charter service to a qualified human service organization (QHSO) for... disabilities; or (3) With low income. (b) If an organization serving persons described in paragraph (a) of this...

  7. 26 CFR 1.1092(c)-1 - Qualified covered calls.

    Science.gov (United States)

    2010-04-01

    ... lowest qualified benchmark is determined using the adjusted applicable stock price, as defined in § 1... (CONTINUED) INCOME TAXES Wash Sales of Stock Or Securities § 1.1092(c)-1 Qualified covered calls. (a) In.... Under section 1092(d)(3)(B)(i)(I), stock is personal property if the stock is part of a straddle that...

  8. 26 CFR 1.860F-1 - Qualified liquidations.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 9 2010-04-01 2010-04-01 false Qualified liquidations. 1.860F-1 Section 1.860F-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Real Estate Investment Trusts § 1.860F-1 Qualified liquidations. A plan of...

  9. 42 CFR 436.128 - Coverage for certain qualified aliens.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 4 2010-10-01 2010-10-01 false Coverage for certain qualified aliens. 436.128... Mandatory Coverage of the Categorically Needy § 436.128 Coverage for certain qualified aliens. The agency... § 440.255(c) of this chapter to those aliens described in § 436.406(c) of this subpart. [55 FR 36820...

  10. 17 CFR 210.12-09 - Valuation and qualifying accounts.

    Science.gov (United States)

    2010-04-01

    ... period Column C—Additions (1)—Charged to costs and expenses (2)—Charged to other accounts—describe Column... qualifying accounts and reserves by descriptive title. Group (a) those valuation and qualifying accounts... accounts. 210.12-09 Section 210.12-09 Commodity and Securities Exchanges SECURITIES AND EXCHANGE COMMISSION...

  11. 49 CFR 192.283 - Plastic pipe: Qualifying joining procedures.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 3 2010-10-01 2010-10-01 false Plastic pipe: Qualifying joining procedures. 192... Materials Other Than by Welding § 192.283 Plastic pipe: Qualifying joining procedures. (a) Heat fusion... for making plastic pipe joints by a heat fusion, solvent cement, or adhesive method, the procedure...

  12. 26 CFR 1.7704-3 - Qualifying income.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 13 2010-04-01 2010-04-01 false Qualifying income. 1.7704-3 Section 1.7704-3 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES General Actuarial Valuations § 1.7704-3 Qualifying income. (a) Certain investment income—(1...

  13. 46 CFR 391.6 - Tax treatment of qualified withdrawals.

    Science.gov (United States)

    2010-10-01

    ... accounting whereby (1) payments shall reduce the basis of the property on the day such payments are actually... 46 Shipping 8 2010-10-01 2010-10-01 false Tax treatment of qualified withdrawals. 391.6 Section...-469 FEDERAL INCOME TAX ASPECTS OF THE CAPITAL CONSTRUCTION FUND § 391.6 Tax treatment of qualified...

  14. 26 CFR 25.2701-4 - Accumulated qualified payments.

    Science.gov (United States)

    2010-04-01

    ... compound interest from the due date of the payment at a rate not less than the appropriate discount rate is... retained interest conferring a distribution right that was previously valued as a qualified payment right (a “qualified payment interest”), the taxable estate or taxable gifts of the individual holding the...

  15. 20 CFR 404.1018b - Medicare qualified government employment.

    Science.gov (United States)

    2010-04-01

    ... AND DISABILITY INSURANCE (1950- ) Employment, Wages, Self-Employment, and Self-Employment Income Work Excluded from Employment § 404.1018b Medicare qualified government employment. (a) General. The work of a... 20 Employees' Benefits 2 2010-04-01 2010-04-01 false Medicare qualified government employment. 404...

  16. 26 CFR 1.405-1 - Qualified bond purchase plans.

    Science.gov (United States)

    2010-04-01

    ... purchase plan must conform to the definition of a pension plan in paragraph (b)(1)(i) of § 1.401-1, or the.... Accordingly, even though a qualified bond purchase plan is designed as a pension plan, it need not provide... apply in a nondiscriminatory manner. (ii) A qualified bond purchase plan which is designed as a pension...

  17. 26 CFR 52.4682-2 - Qualifying sales.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 17 2010-04-01 2010-04-01 false Qualifying sales. 52.4682-2 Section 52.4682-2... TAXES (CONTINUED) ENVIRONMENTAL TAXES § 52.4682-2 Qualifying sales. (a) In general—(1) Special rules applicable to certain sales. Special rules apply to sales of ODCs in the following cases: (i) Under section...

  18. The qualified possession turn into ownership

    Directory of Open Access Journals (Sweden)

    Popov Danica

    2011-01-01

    Full Text Available Possession is prima facie evidence of ownership. Possession is ninetents of the law, means that possession is good against all other, except the true owner. The possession ripens into ownership if it is qualified and by effluxion of time. In Serbian law there are two kinds of adverse possession ripens into ownership. The first one is named ordinary and second one extraordinary adverse possession. Ordinary possession need to be legal, conscientious and genuine. Extraordinary possession is only conscientious, but in a wide sense. Adverse possession destroys the title of the owner and vests it in possessor. An occupation of land inconsistent with the right of the true owner: the possession of those against whom a right action has accured to the true owner. It is actual possession in the absence of possession by the rightful owner and without lawful title. If the adverse possession continues, the effect at the expiration of the prescribed period is that not only the remedy but the title of former owner is extinguished. The person in adverse possession gains a new possessory title which cannot, normally exceed in extent of duration the interest of the former owner.

  19. Intracellular localization of Arabidopsis sulfurtransferases.

    Science.gov (United States)

    Bauer, Michael; Dietrich, Christof; Nowak, Katharina; Sierralta, Walter D; Papenbrock, Jutta

    2004-06-01

    Sulfurtransferases (Str) comprise a group of enzymes widely distributed in archaea, eubacteria, and eukaryota which catalyze the transfer of a sulfur atom from suitable sulfur donors to nucleophilic sulfur acceptors. In all organisms analyzed to date, small gene families encoding Str proteins have been identified. The gene products were localized to different compartments of the cells. Our interest concerns the localization of Str proteins encoded in the nuclear genome of Arabidopsis. Computer-based prediction methods revealed localization in different compartments of the cell for six putative AtStrs. Several methods were used to determine the localization of the AtStr proteins experimentally. For AtStr1, a mitochondrial localization was demonstrated by immunodetection in the proteome of isolated mitochondria resolved by one- and two-dimensional gel electrophoresis and subsequent blotting. The respective mature AtStr1 protein was identified by mass spectrometry sequencing. The same result was obtained by transient expression of fusion constructs with the green fluorescent protein in Arabidopsis protoplasts, whereas AtStr2 was exclusively localized to the cytoplasm by this method. Three members of the single-domain AtStr were localized in the chloroplasts as demonstrated by transient expression of green fluorescent protein fusions in protoplasts and stomata, whereas the single-domain AtStr18 was shown to be cytoplasmic. The remarkable subcellular distribution of AtStr15 was additionally analyzed by transmission electron immunomicroscopy using a monospecific antibody against green fluorescent protein, indicating an attachment to the thylakoid membrane. The knowledge of the intracellular localization of the members of this multiprotein family will help elucidate their specific functions in the organism.

  20. Intracellular calcium homeostasis and signaling.

    Science.gov (United States)

    Brini, Marisa; Calì, Tito; Ottolini, Denis; Carafoli, Ernesto

    2013-01-01

    Ca(2+) is a universal carrier of biological information: it controls cell life from its origin at fertilization to its end in the process of programmed cell death. Ca(2+) is a conventional diffusible second messenger released inside cells by the interaction of first messengers with plasma membrane receptors. However, it can also penetrate directly into cells to deliver information without the intermediation of first or second messengers. Even more distinctively, Ca(2+) can act as a first messenger, by interacting with a plasma membrane receptor to set in motion intracellular signaling pathways that involve Ca(2+) itself. Perhaps the most distinctive property of the Ca(2+) signal is its ambivalence: while essential to the correct functioning of cells, Ca(2+) becomes an agent that mediates cell distress, or even (toxic) cell death, if its concentration and movements inside cells are not carefully tuned. Ca(2+) is controlled by reversible complexation to specific proteins, which could be pure Ca(2+) buffers, or which, in addition to buffering Ca(2+), also decode its signal to pass it on to targets. The most important actors in the buffering of cell Ca(2+) are proteins that transport it across the plasma membrane and the membrane of the organelles: some have high Ca(2+) affinity and low transport capacity (e.g., Ca(2+) pumps), others have opposite properties (e.g., the Ca(2+) uptake system of mitochondria). Between the initial event of fertilization, and the terminal event of programmed cell death, the Ca(2+) signal regulates the most important activities of the cell, from the expression of genes, to heart and muscle contraction and other motility processes, to diverse metabolic pathways involved in the generation of cell fuels.

  1. 11 CFR 9004.4 - Use of payments; examples of qualified campaign expenses and non-qualified campaign expenses.

    Science.gov (United States)

    2010-01-01

    ... 11 Federal Elections 1 2010-01-01 2010-01-01 false Use of payments; examples of qualified campaign expenses and non-qualified campaign expenses. 9004.4 Section 9004.4 Federal Elections FEDERAL ELECTION COMMISSION PRESIDENTIAL ELECTION CAMPAIGN FUND: GENERAL ELECTION FINANCING ENTITLEMENT OF ELIGIBLE CANDIDATES...

  2. Nanoparticles for intracellular-targeted drug delivery

    International Nuclear Information System (INIS)

    Paulo, Cristiana S O; Pires das Neves, Ricardo; Ferreira, Lino S

    2011-01-01

    Nanoparticles (NPs) are very promising for the intracellular delivery of anticancer and immunomodulatory drugs, stem cell differentiation biomolecules and cell activity modulators. Although initial studies in the area of intracellular drug delivery have been performed in the delivery of DNA, there is an increasing interest in the use of other molecules to modulate cell activity. Herein, we review the latest advances in the intracellular-targeted delivery of short interference RNA, proteins and small molecules using NPs. In most cases, the drugs act at different cellular organelles and therefore the drug-containing NPs should be directed to precise locations within the cell. This will lead to the desired magnitude and duration of the drug effects. The spatial control in the intracellular delivery might open new avenues to modulate cell activity while avoiding side-effects.

  3. Biological synthesis and characterization of intracellular gold ...

    Indian Academy of Sciences (India)

    thods of reduction of metal ions using plants or microorganisms are often ... have several advantages over bacteria, they are often pre- ferred. ... in static condition for a period of 7 days. ... work was focused on the production of intracellular gold.

  4. Mycobacterium intracellulare Infection Mimicking Progression of Scleroderma

    DEFF Research Database (Denmark)

    Krabbe, Simon; Engelhart, Merete; Thybo, Sören

    2017-01-01

    This case report describes a patient with scleroderma who developed Mycobacterium intracellulare infection, which for more than a year mimicked worsening of her connective tissue disorder. The patient was diagnosed with scleroderma based on puffy fingers that developed into sclerodactyly, abnormal......, unfortunately with significant scarring. Immunodeficiency testing was unremarkable. In summary, an infection with Mycobacterium intracellulare was mistaken for an unusually severe progression of scleroderma....

  5. Supporting Qualified Database for Uncertainty Evaluation

    International Nuclear Information System (INIS)

    Petruzzi, A.; Fiori, F.; Kovtonyuk, A.; Lisovyy, O.; D'Auria, F.

    2013-01-01

    Uncertainty evaluation constitutes a key feature of BEPU (Best Estimate Plus Uncertainty) process. The uncertainty can be the result of a Monte Carlo type analysis involving input uncertainty parameters or the outcome of a process involving the use of experimental data and connected code calculations. Those uncertainty methods are discussed in several papers and guidelines (IAEA-SRS-52, OECD/NEA BEMUSE reports). The present paper aims at discussing the role and the depth of the analysis required for merging from one side suitable experimental data and on the other side qualified code calculation results. This aspect is mostly connected with the second approach for uncertainty mentioned above, but it can be used also in the framework of the first approach. Namely, the paper discusses the features and structure of the database that includes the following kinds of documents: 1. The 'RDS-facility' (Reference Data Set for the selected facility): this includes the description of the facility, the geometrical characterization of any component of the facility, the instrumentations, the data acquisition system, the evaluation of pressure losses, the physical properties of the material and the characterization of pumps, valves and heat losses; 2. The 'RDS-test' (Reference Data Set for the selected test of the facility): this includes the description of the main phenomena investigated during the test, the configuration of the facility for the selected test (possible new evaluation of pressure and heat losses if needed) and the specific boundary and initial conditions; 3. The 'QP' (Qualification Report) of the code calculation results: this includes the description of the nodalization developed following a set of homogeneous techniques, the achievement of the steady state conditions and the qualitative and quantitative analysis of the transient with the characterization of the Relevant Thermal-Hydraulics Aspects (RTA); 4. The EH (Engineering Handbook) of the input nodalization

  6. Supporting qualified database for uncertainty evaluation

    Energy Technology Data Exchange (ETDEWEB)

    Petruzzi, A.; Fiori, F.; Kovtonyuk, A.; D' Auria, F. [Nuclear Research Group of San Piero A Grado, Univ. of Pisa, Via Livornese 1291, 56122 Pisa (Italy)

    2012-07-01

    Uncertainty evaluation constitutes a key feature of BEPU (Best Estimate Plus Uncertainty) process. The uncertainty can be the result of a Monte Carlo type analysis involving input uncertainty parameters or the outcome of a process involving the use of experimental data and connected code calculations. Those uncertainty methods are discussed in several papers and guidelines (IAEA-SRS-52, OECD/NEA BEMUSE reports). The present paper aims at discussing the role and the depth of the analysis required for merging from one side suitable experimental data and on the other side qualified code calculation results. This aspect is mostly connected with the second approach for uncertainty mentioned above, but it can be used also in the framework of the first approach. Namely, the paper discusses the features and structure of the database that includes the following kinds of documents: 1. The' RDS-facility' (Reference Data Set for the selected facility): this includes the description of the facility, the geometrical characterization of any component of the facility, the instrumentations, the data acquisition system, the evaluation of pressure losses, the physical properties of the material and the characterization of pumps, valves and heat losses; 2. The 'RDS-test' (Reference Data Set for the selected test of the facility): this includes the description of the main phenomena investigated during the test, the configuration of the facility for the selected test (possible new evaluation of pressure and heat losses if needed) and the specific boundary and initial conditions; 3. The 'QR' (Qualification Report) of the code calculation results: this includes the description of the nodalization developed following a set of homogeneous techniques, the achievement of the steady state conditions and the qualitative and quantitative analysis of the transient with the characterization of the Relevant Thermal-Hydraulics Aspects (RTA); 4. The EH (Engineering

  7. Aging techniques and qualified life for safety system components

    International Nuclear Information System (INIS)

    Weaver, W.W.

    1980-01-01

    Presently, the qualified life objective for Class IE safety system components in nuclear power plants is somewhat of a subjective engineering judgment. When the desired qualified life is ascertained, there are other choices that must be made (which may be influenced by the desired qualified life) such as selecting the aging procedure to use in the qualification process. Adding complexity to the situation is the fact that there are some limitations in aging techniques at the present time. This article presents (1) a discussion of the limitations in aging procedures, (2) the general philosophy of qualification, and (3) a proposed method for specifying a desired qualified life, which uses a probabilistic approach. The probabilistic approach proposed in item 3 can be applied to natural aging programs and eventually to accelerated aging once the present technical difficulties are overcome

  8. The technical qualified expert on radiation protection in Spain

    International Nuclear Information System (INIS)

    Marco Arboli, M.; Rodriguez Suarez, M.

    2002-01-01

    Education and training is considered an important tool for promoting safety culture and improving the level of competence of workers. In the different disciplines involved in Radiation Protection (PR), training programmes are being revised to ensure effective protection of individuals. In the European Union framework, the new normative and its adaptation in the Member States, as well as the past actions in each country, make the community to conclude that harmonization of educational programmes and European recognition of the qualification must be reached. An important point of these actions is the definition of the European Qualified Expert. Article 38 of the EURATOM Basic Safety Standards imposes requirements on training and education of the qualified experts and their exchange within the European Union. Since then, there has been many initiatives to seek to harmonization of the qualified expert requirements. In the Spanish education system, there has been a standard training and competent authority recognition of the high level qualified experts on radiation protection since 1986, whose programme is continuously being updating and improving. Taking into account the European actions to define the qualified expert training requirements, it has been designed an educational course for the Technical Qualified Expert on Radiation Protection. This pilot course has being carried out during May, 2002. The results of this project is shown in this paper. (Author)

  9. Efficient intracellular delivery and improved biocompatibility of colloidal silver nanoparticles towards intracellular SERS immuno-sensing.

    Science.gov (United States)

    Bhardwaj, Vinay; Srinivasan, Supriya; McGoron, Anthony J

    2015-06-21

    High throughput intracellular delivery strategies, electroporation, passive and TATHA2 facilitated diffusion of colloidal silver nanoparticles (AgNPs) are investigated for cellular toxicity and uptake using state-of-art analytical techniques. The TATHA2 facilitated approach efficiently delivered high payload with no toxicity, pre-requisites for intracellular applications of plasmonic metal nanoparticles (PMNPs) in sensing and therapeutics.

  10. Investigating Internalization and Intracellular Trafficking of GPCRs

    DEFF Research Database (Denmark)

    Foster, Simon R; Bräuner-Osborne, Hans

    2017-01-01

    for signal transduction. One of the major mechanisms for GPCR regulation involves their endocytic trafficking, which serves to internalize the receptors from the plasma membrane and thereby attenuate G protein-dependent signaling. However, there is accumulating evidence to suggest that GPCRs can signal...... independently of G proteins, as well as from intracellular compartments including endosomes. It is in this context that receptor internalization and intracellular trafficking have attracted renewed interest within the GPCR field. In this chapter, we will review the current understanding and methodologies...

  11. Cationic polymers for intracellular delivery of proteins

    NARCIS (Netherlands)

    Coué, G.M.J.P.C.; Engbersen, Johannes F.J.; Samal, Sangram; Dubruel, Peter

    2015-01-01

    Many therapeutic proteins exert their pharmaceutical action inside the cytoplasm or onto individual organelles inside the cell. Intracellular protein delivery is considered to be the most direct, fastest and safest approach for curing gene-deficiency diseases, enhancing vaccination and triggering

  12. Molecular detection and characterization of sustainable intracellular ...

    African Journals Online (AJOL)

    3Centre for Biopolymer and Bio-Molecular Research, Athlone College of Technology, Republic of Ireland. ... cells was associated with the elongation of micro-villar extension that ... Keywords: Intracellular contaminants, cell cultures, bacteria culture, pre-clinical studies. ... production work involving culture technology.

  13. Spatial Cytoskeleton Organization Supports Targeted Intracellular Transport

    Science.gov (United States)

    Hafner, Anne E.; Rieger, Heiko

    2018-03-01

    The efficiency of intracellular cargo transport from specific source to target locations is strongly dependent upon molecular motor-assisted motion along the cytoskeleton. Radial transport along microtubules and lateral transport along the filaments of the actin cortex underneath the cell membrane are characteristic for cells with a centrosome. The interplay between the specific cytoskeleton organization and the motor performance realizes a spatially inhomogeneous intermittent search strategy. In order to analyze the efficiency of such intracellular search strategies we formulate a random velocity model with intermittent arrest states. We evaluate efficiency in terms of mean first passage times for three different, frequently encountered intracellular transport tasks: i) the narrow escape problem, which emerges during cargo transport to a synapse or other specific region of the cell membrane, ii) the reaction problem, which considers the binding time of two particles within the cell, and iii) the reaction-escape problem, which arises when cargo must be released at a synapse only after pairing with another particle. Our results indicate that cells are able to realize efficient search strategies for various intracellular transport tasks economically through a spatial cytoskeleton organization that involves only a narrow actin cortex rather than a cell body filled with randomly oriented actin filaments.

  14. Biological synthesis and characterization of intracellular gold ...

    Indian Academy of Sciences (India)

    In the present study, Aspergillus fumigatus was used for the intracellular synthesis of gold nanoparticles. Stable nanoparticles were produced when an aqueous solution of chloroauric acid (HAuCl4) was reduced by A. fumigatus biomass as the reducing agent. Production of nanoparticles was confirmed by the colour ...

  15. Optimizing Nanoelectrode Arrays for Scalable Intracellular Electrophysiology.

    Science.gov (United States)

    Abbott, Jeffrey; Ye, Tianyang; Ham, Donhee; Park, Hongkun

    2018-03-20

    Electrode technology for electrophysiology has a long history of innovation, with some decisive steps including the development of the voltage-clamp measurement technique by Hodgkin and Huxley in the 1940s and the invention of the patch clamp electrode by Neher and Sakmann in the 1970s. The high-precision intracellular recording enabled by the patch clamp electrode has since been a gold standard in studying the fundamental cellular processes underlying the electrical activities of neurons and other excitable cells. One logical next step would then be to parallelize these intracellular electrodes, since simultaneous intracellular recording from a large number of cells will benefit the study of complex neuronal networks and will increase the throughput of electrophysiological screening from basic neurobiology laboratories to the pharmaceutical industry. Patch clamp electrodes, however, are not built for parallelization; as for now, only ∼10 patch measurements in parallel are possible. It has long been envisioned that nanoscale electrodes may help meet this challenge. First, nanoscale electrodes were shown to enable intracellular access. Second, because their size scale is within the normal reach of the standard top-down fabrication, the nanoelectrodes can be scaled into a large array for parallelization. Third, such a nanoelectrode array can be monolithically integrated with complementary metal-oxide semiconductor (CMOS) electronics to facilitate the large array operation and the recording of the signals from a massive number of cells. These are some of the central ideas that have motivated the research activity into nanoelectrode electrophysiology, and these past years have seen fruitful developments. This Account aims to synthesize these findings so as to provide a useful reference. Summing up from the recent studies, we will first elucidate the morphology and associated electrical properties of the interface between a nanoelectrode and a cellular membrane

  16. 26 CFR 1.42-6 - Buildings qualifying for carryover allocations.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 1 2010-04-01 2010-04-01 true Buildings qualifying for carryover allocations. 1... INCOME TAXES Credits Against Tax § 1.42-6 Buildings qualifying for carryover allocations. (a) Carryover... carryover allocation may only be made with respect to a qualified building. A qualified building is any...

  17. 26 CFR 48.4041-19 - Exemption for qualified methanol and ethanol fuel.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 16 2010-04-01 2010-04-01 true Exemption for qualified methanol and ethanol....4041-19 Exemption for qualified methanol and ethanol fuel. (a) In general. Under section 4041(b)(2... or use of qualified methanol or ethanol fuel. (b) Qualified methanol or ethanol fuel defined. For...

  18. 26 CFR 1.43-4 - Qualified enhanced oil recovery costs.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 1 2010-04-01 2010-04-01 true Qualified enhanced oil recovery costs. 1.43-4... TAXES Credits Against Tax § 1.43-4 Qualified enhanced oil recovery costs. (a) Qualifying costs—(1) In... “qualified enhanced oil recovery costs” if the amounts are paid or incurred with respect to an asset which is...

  19. Qualified operator training in the simulated control room environment

    International Nuclear Information System (INIS)

    Ionescu, Teodor; Studineanu, Emil; Radulescu, Catalina; Bolocan, Gabriel

    2005-01-01

    Full text: Mainly designed for the training of the Cernavoda NPP Unit 2 operators, the virtual simulated environment allows the training of the already qualified operators for Cernavoda NPP Unit 1, adding to the already trained knowledge, the differences which has occurred in the Unit 2 design. Using state-of-the-art computers and displays and qualified software, the virtual simulated panels could offer a viable alternative to classic hardware-based training. This approach allows quick training of the new procedures required by the new configuration of the re-designed operator panels in the main control room of Cernavoda NPP Unit 2. (authors)

  20. Qualified operator training in the simulated control room environment

    International Nuclear Information System (INIS)

    Ionescu, Teodor; Studineanu, Emil; Radulescu, Catalina; Bolocan, Gabriel

    2005-01-01

    Mainly designed for the training of the Cernavoda NPP Unit 2 operators, the virtual simulated environment allows the training of the already qualified operators for Cernavoda NPP Unit 1, adding to the already trained knowledge, the differences which have occurred in the Unit 2 design. Using state-of-the-art computers and displays and qualified software, the virtual simulated panels could offer a viable alternative to classic hardware-based training. This approach allows quick training of the new procedures required by the new configuration of the re-designed operator panels in the main control room of Cernavoda NPP Unit 2. (authors)

  1. Accident data study of concrete construction companies' similarities and differences between qualified and non-qualified workers in Spain.

    Science.gov (United States)

    López-Arquillos, Antonio; Rubio-Romero, Juan Carlos; Gibb, Alistair

    2015-01-01

    The aim of this paper is to discuss findings from an analysis of accidents in concrete construction companies in Spain and to compare the accident rates of qualified and non-qualified workers. A total of 125,021 accidents between 2003 and 2008 involving both blue-collar and white-collar workers were analysed, comparing the variables of occupation, age, company staff, length of service, location of the accident, together with the severity of the accidents. Results showed that lack of experience in the first month is more significant in non-qualified workers and experienced supervisors and that head injuries are more likely to lead to fatalities. The most remarkable similarity was that fatal accidents to and from the worksite are a problem common to both groups of workers.

  2. 31 CFR Appendix A to Part 500 - Qualifying International Institutions

    Science.gov (United States)

    2010-07-01

    ... Pt. 500, App. A Appendix A to Part 500—Qualifying International Institutions Asian Development Bank (ADB) Food and Agricultural Organization (FAO) International Bank for Reconstruction and Development (IBRD, the “World Bank”) International Civil Aviation Organization (ICAO) International Development...

  3. 77 FR 26175 - Section 42 Qualified Contract Provisions

    Science.gov (United States)

    2012-05-03

    ... indebtedness under general principles of Federal income tax law, and (3) upon the sale of the building, are... building for sale to the general public at the determined qualified contract price upon receipt of a... because of variations under State laws concerning the terms of a bona fide contract and methods for...

  4. 77 FR 9846 - Source of Income From Qualified Fails Charges

    Science.gov (United States)

    2012-02-21

    ... temporary regulations noted that no trading practice existed at that time for fails charges on securities other than Treasuries, but that if a fails charge trading practice pertaining to other securities was... sources within the United States, and the income from the qualified fails charge is treated as effectively...

  5. 9 CFR 77.36 - Interstate movement from qualified herds.

    Science.gov (United States)

    2010-01-01

    ... date of movement. If a group of captive cervids from a qualified herd is being moved interstate... unclassified herd. (4) Captive cervids being moved interstate for the purpose of exhibition only may be moved... being moved directly from a classified herd, the captive cervid must be isolated from all other animals...

  6. Validation of mentorship model for newly qualified professional ...

    African Journals Online (AJOL)

    Newly qualified professional nurses (NQPNs) allocated to community health care services require the use of validated model to practice independently. Validation was done to adapt and assess if the model is understood and could be implemented by NQPNs and mentors employed in community health care services.

  7. 36 CFR 908.13 - Rights of Qualified Persons.

    Science.gov (United States)

    2010-07-01

    ... POLICY AND PROCEDURES TO FACILITATE THE RETENTION OF DISPLACED BUSINESSES AND RESIDENTS IN THE... leasing plan which are, in the Corporation's opinion, major. (c) Each Qualified Person on the List, who is... opportunity to occupy Newly Developed Space shall not be limited to the square on which its previous location...

  8. 42 CFR 416.26 - Qualifying for an agreement.

    Science.gov (United States)

    2010-10-01

    ..., where State law requires licensure, the ASC complies with State licensure requirements; and (3) The ASC.... (c) Acceptance of the ASC as qualified to furnish ambulatory surgical services. If CMS determines... its authorized representative; and (2) File them with CMS. (e) Acceptance by CMS. If CMS accepts the...

  9. 22 CFR 19.6-2 - Qualifying court order.

    Science.gov (United States)

    2010-04-01

    ... principal's Foreign Service retirement benefit or survivor benefit. If a court directs or implies that a... Foreign Relations DEPARTMENT OF STATE PERSONNEL BENEFITS FOR SPOUSES AND FORMER SPOUSES OF PARTICIPANTS IN... the Secretary of State. A qualifying court order must— (1) Be consistent with the terms of the Act and...

  10. 34 CFR 300.18 - Highly qualified special education teachers.

    Science.gov (United States)

    2010-07-01

    ... 34 Education 2 2010-07-01 2010-07-01 false Highly qualified special education teachers. 300.18... SPECIAL EDUCATION AND REHABILITATIVE SERVICES, DEPARTMENT OF EDUCATION ASSISTANCE TO STATES FOR THE... special education teachers. (a) Requirements for special education teachers teaching core academic...

  11. 24 CFR 266.100 - Qualified housing finance agency (HFA).

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 2 2010-04-01 2010-04-01 false Qualified housing finance agency (HFA). 266.100 Section 266.100 Housing and Urban Development Regulations Relating to Housing and Urban Development (Continued) OFFICE OF ASSISTANT SECRETARY FOR HOUSING-FEDERAL HOUSING COMMISSIONER, DEPARTMENT OF HOUSING AND URBAN DEVELOPMENT MORTGAGE...

  12. 26 CFR 1.465-27 - Qualified nonrecourse financing.

    Science.gov (United States)

    2010-04-01

    ... partnership; incidental property. (i) X is a limited liability company that is classified as a partnership for.... (i) UTP1 and UTP2, both limited liability companies classified as partnerships, are the only general... as qualified nonrecourse financing. (4) Partnership liability. For purposes of section 465(b)(6) and...

  13. 26 CFR 54.4980B-3 - Qualified beneficiaries.

    Science.gov (United States)

    2010-04-01

    ... the day before the qualifying event (that is, the divorce or death) will have been by reason of D's... event, and the reason for the individual's lack of actual coverage (such as the individual's having..., the individual is covered under the group health plan by reason of another individual's election of...

  14. 30 CFR 75.153 - Electrical work; qualified person.

    Science.gov (United States)

    2010-07-01

    ... Mine Safety and Health Districts at regular intervals, or as demand requires. (d) A score of at least... Section 75.153 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR COAL MINE SAFETY AND HEALTH MANDATORY SAFETY STANDARDS-UNDERGROUND COAL MINES Qualified and Certified Persons § 75...

  15. 26 CFR 1.41-2 - Qualified research expenses.

    Science.gov (United States)

    2010-04-01

    ... Credits Against Tax § 1.41-2 Qualified research expenses. (a) Trade or business requirement—(1) In general... research and experimental expenses) are not necessarily paid or incurred in carrying on a trade or business for purposes of section 41. A research expense must relate to a particular trade or business being...

  16. Dynamics of gradient formation by intracellular shuttling

    Energy Technology Data Exchange (ETDEWEB)

    Berezhkovskii, Alexander M. [Mathematical and Statistical Computing Laboratory, Division of Computational Bioscience, Center for Information Technology, National Institutes of Health, Bethesda, Maryland 20892 (United States); Shvartsman, Stanislav Y. [Department of Chemical and Biological Engineering and Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey 08544 (United States)

    2015-08-21

    A number of important cellular functions rely on the formation of intracellular protein concentration gradients. Experimental studies discovered a number of mechanisms for the formation of such gradients. One of the mechanisms relies on the intracellular shuttling of a protein that interconverts between the two states with different diffusivities, under the action of two enzymes, one of which is localized to the plasma membrane, whereas the second is uniformly distributed in the cytoplasm. Recent work reported an analytical solution for the steady state gradient in this mechanism, obtained in the framework of a one-dimensional reaction-diffusion model. Here, we study the dynamics in this model and derive analytical expressions for the Laplace transforms of the time-dependent concentration profiles in terms of elementary transcendental functions. Inverting these transforms numerically, one can obtain time-dependent concentration profiles of the two forms of the protein.

  17. Leishmania hijacking of the macrophage intracellular compartments.

    Science.gov (United States)

    Liévin-Le Moal, Vanessa; Loiseau, Philippe M

    2016-02-01

    Leishmania spp., transmitted to humans by the bite of the sandfly vector, are responsible for the three major forms of leishmaniasis, cutaneous, diffuse mucocutaneous and visceral. Leishmania spp. interact with membrane receptors of neutrophils and macrophages. In macrophages, the parasite is internalized within a parasitophorous vacuole and engages in a particular intracellular lifestyle in which the flagellated, motile Leishmania promastigote metacyclic form differentiates into non-motile, metacyclic amastigote form. This phenomenon is induced by Leishmania-triggered events leading to the fusion of the parasitophorous vacuole with vesicular members of the host cell endocytic pathway including recycling endosomes, late endosomes and the endoplasmic reticulum. Maturation of the parasitophorous vacuole leads to the intracellular proliferation of the Leishmania amastigote forms by acquisition of host cell nutrients while escaping host defense responses. © 2015 FEBS.

  18. Reduction of intracellular glutathione content and radiosensitivity

    International Nuclear Information System (INIS)

    Vos, O.; Schans, G.P. van der; Roos-Verheij, W.S.D.

    1986-05-01

    The intracellular glutathione (GSH) content in HeLa, CHO and V79 cells was reduced by incubating the cells in growth medium containing buthionine sulfoximine (BSO) or diethyl maleate (DEM). Clonogenicity, single strand DNA breaks (ssb) and double strand DNA breaks (dsb) were used as criteria for radiation induced damage after X- or γ irradiation. In survival experiments DEM gave a slightly larger sensitization although it gave a smaller reduction of the intracellular GSH. In general, sensitization was larger for dsb than for ssb, also the reduction of the OER was generally larger for dsb than for ssb. This may be due to the higher dose rate in case of dsb experiments resulting in a higher rate of radiochemical oxygen consumption. In general, no effect was found on post-irradiation repair of ssb and dsb. (Auth.)

  19. Reduction of intracellular glutathione content and radiosensitivity

    International Nuclear Information System (INIS)

    Vos, O.; Schans, G.P. van der; Roos-Verheij, W.S.D.

    1986-01-01

    The intracellular glutathione (GSH) content of HeLa, CHO and V79 cells was reduced by incubating the cells in growth medium containing buthionine sulphoximine or diethyl maleate (DEM). Clonogenicity, single-strand DNA breaks (ssb) and double-strand DNA breaks (dsb) were used as criteria for radiation-induced damage after X- or γ-irradiation. In survival experiments, DEM gave a slightly larger sensitization although it gave a smaller reduction of the intracellular GSH. In general, sensitization was larger for dsb than for ssb, also the reduction of the o.e.r. was generally larger for dsb than for ssb. This may be due to the higher dose rate in case of dsb experiments resulting in a higher rate of radiochemical oxygen consumption. In general, no effect was found on post-irradiation repair of ssb and dsb. (author)

  20. Intracellular mechanisms of solar water disinfection

    Science.gov (United States)

    Castro-Alférez, María; Polo-López, María Inmaculada; Fernández-Ibáñez, Pilar

    2016-12-01

    Solar water disinfection (SODIS) is a zero-cost intervention measure to disinfect drinking water in areas of poor access to improved water sources, used by more than 6 million people in the world. The bactericidal action of solar radiation in water has been widely proven, nevertheless the causes for this remain still unclear. Scientific literature points out that generation of reactive oxygen species (ROS) inside microorganisms promoted by solar light absorption is the main reason. For the first time, this work reports on the experimental measurement of accumulated intracellular ROS in E. coli during solar irradiation. For this experimental achievement, a modified protocol based on the fluorescent probe dichlorodihydrofluorescein diacetate (DCFH-DA), widely used for oxidative stress in eukaryotic cells, has been tested and validated for E. coli. Our results demonstrate that ROS and their accumulated oxidative damages at intracellular level are key in solar water disinfection.

  1. Intracellular serpins, firewalls and tissue necrosis.

    Science.gov (United States)

    Marciniak, Stefan J; Lomas, David A

    2008-02-01

    Luke and colleagues have recently attributed a new role to a member of the serpin superfamily of serine proteinase inhibitors. They have used Caenorhabditis elegans to show that an intracellular serpin is crucial for maintaining lysosomal integrity. We examine the role of this firewall in preventing necrosis and attempt to integrate this with current theories of stress-induced protein degradation. We discuss how mutant serpins cause disease either through polymerization or now, perhaps, by unleashing necrosis.

  2. Fluorescent nanoparticles for intracellular sensing: A review

    International Nuclear Information System (INIS)

    Ruedas-Rama, Maria J.; Walters, Jamie D.; Orte, Angel; Hall, Elizabeth A.H.

    2012-01-01

    Highlights: ► Analytical applications of fluorescent nanoparticles (NPs) in intracellular sensing. ► Critical review on performance of QDots, metal NPs, silica NPs, and polymer NPs. ► Highlighted potential of fluorescence lifetime imaging microscopy (FLIM). - Abstract: Fluorescent nanoparticles (NPs), including semiconductor NPs (Quantum Dots), metal NPs, silica NPs, polymer NPs, etc., have been a major focus of research and development during the past decade. The fluorescent nanoparticles show unique chemical and optical properties, such as brighter fluorescence, higher photostability and higher biocompatibility, compared to classical fluorescent organic dyes. Moreover, the nanoparticles can also act as multivalent scaffolds for the realization of supramolecular assemblies, since their high surface to volume ratio allow distinct spatial domains to be functionalized, which can provide a versatile synthetic platform for the implementation of different sensing schemes. Their excellent properties make them one of the most useful tools that chemistry has supplied to biomedical research, enabling the intracellular monitoring of many different species for medical and biological purposes. In this review, we focus on the developments and analytical applications of fluorescent nanoparticles in chemical and biological sensing within the intracellular environment. The review also points out the great potential of fluorescent NPs for fluorescence lifetime imaging microscopy (FLIM). Finally, we also give an overview of the current methods for delivering of fluorescent NPs into cells, where critically examine the benefits and liabilities of each strategy.

  3. Fluorescent nanoparticles for intracellular sensing: A review

    Energy Technology Data Exchange (ETDEWEB)

    Ruedas-Rama, Maria J., E-mail: mjruedas@ugr.esmailto [Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Campus Cartuja, 18071, Granada (Spain); Walters, Jamie D. [Department of Chemical Engineering and Biotechnology, University of Cambridge, Tennis Court Road, Cambridge, UK CB2 1QT (United Kingdom); Orte, Angel [Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Campus Cartuja, 18071, Granada (Spain); Hall, Elizabeth A.H., E-mail: lisa.hall@biotech.cam.ac.uk [Department of Chemical Engineering and Biotechnology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QT (United Kingdom)

    2012-11-02

    Highlights: Black-Right-Pointing-Pointer Analytical applications of fluorescent nanoparticles (NPs) in intracellular sensing. Black-Right-Pointing-Pointer Critical review on performance of QDots, metal NPs, silica NPs, and polymer NPs. Black-Right-Pointing-Pointer Highlighted potential of fluorescence lifetime imaging microscopy (FLIM). - Abstract: Fluorescent nanoparticles (NPs), including semiconductor NPs (Quantum Dots), metal NPs, silica NPs, polymer NPs, etc., have been a major focus of research and development during the past decade. The fluorescent nanoparticles show unique chemical and optical properties, such as brighter fluorescence, higher photostability and higher biocompatibility, compared to classical fluorescent organic dyes. Moreover, the nanoparticles can also act as multivalent scaffolds for the realization of supramolecular assemblies, since their high surface to volume ratio allow distinct spatial domains to be functionalized, which can provide a versatile synthetic platform for the implementation of different sensing schemes. Their excellent properties make them one of the most useful tools that chemistry has supplied to biomedical research, enabling the intracellular monitoring of many different species for medical and biological purposes. In this review, we focus on the developments and analytical applications of fluorescent nanoparticles in chemical and biological sensing within the intracellular environment. The review also points out the great potential of fluorescent NPs for fluorescence lifetime imaging microscopy (FLIM). Finally, we also give an overview of the current methods for delivering of fluorescent NPs into cells, where critically examine the benefits and liabilities of each strategy.

  4. Intracellular Cholesterol Trafficking and Impact in Neurodegeneration

    Directory of Open Access Journals (Sweden)

    Fabian Arenas

    2017-11-01

    Full Text Available Cholesterol is a critical component of membrane bilayers where it plays key structural and functional roles by regulating the activity of diverse signaling platforms and pathways. Particularly enriched in brain, cholesterol homeostasis in this organ is singular with respect to other tissues and exhibits a heterogeneous regulation in distinct brain cell populations. Due to the key role of cholesterol in brain physiology and function, alterations in cholesterol homeostasis and levels have been linked to brain diseases and neurodegeneration. In the case of Alzheimer disease (AD, however, this association remains unclear with evidence indicating that either increased or decreased total brain cholesterol levels contribute to this major neurodegenerative disease. Here, rather than analyzing the role of total cholesterol levels in neurodegeneration, we focus on the contribution of intracellular cholesterol pools, particularly in endolysosomes and mitochondria through its trafficking via specialized membrane domains delineated by the contacts between endoplasmic reticulum and mitochondria, in the onset of prevalent neurodegenerative diseases such as AD, Parkinson disease, and Huntington disease as well as in lysosomal disorders like Niemann-Pick type C disease. We dissect molecular events associated with intracellular cholesterol accumulation, especially in mitochondria, an event that results in impaired mitochondrial antioxidant defense and function. A better understanding of the mechanisms involved in the distribution of cholesterol in intracellular compartments may shed light on the role of cholesterol homeostasis disruption in neurodegeneration and may pave the way for specific intervention opportunities.

  5. A bacteriophage endolysin that eliminates intracellular streptococci

    Science.gov (United States)

    Shen, Yang; Barros, Marilia; Vennemann, Tarek; Gallagher, D Travis; Yin, Yizhou; Linden, Sara B; Heselpoth, Ryan D; Spencer, Dennis J; Donovan, David M; Moult, John; Fischetti, Vincent A; Heinrich, Frank; Lösche, Mathias; Nelson, Daniel C

    2016-01-01

    PlyC, a bacteriophage-encoded endolysin, lyses Streptococcus pyogenes (Spy) on contact. Here, we demonstrate that PlyC is a potent agent for controlling intracellular Spy that often underlies refractory infections. We show that the PlyC holoenzyme, mediated by its PlyCB subunit, crosses epithelial cell membranes and clears intracellular Spy in a dose-dependent manner. Quantitative studies using model membranes establish that PlyCB interacts strongly with phosphatidylserine (PS), whereas its interaction with other lipids is weak, suggesting specificity for PS as its cellular receptor. Neutron reflection further substantiates that PlyC penetrates bilayers above a PS threshold concentration. Crystallography and docking studies identify key residues that mediate PlyCB–PS interactions, which are validated by site-directed mutagenesis. This is the first report that a native endolysin can traverse epithelial membranes, thus substantiating the potential of PlyC as an antimicrobial for Spy in the extracellular and intracellular milieu and as a scaffold for engineering other functionalities. DOI: http://dx.doi.org/10.7554/eLife.13152.001 PMID:26978792

  6. Competence of newly qualified registered nurses from a nursing college

    Directory of Open Access Journals (Sweden)

    BG Morolong

    2005-09-01

    Full Text Available The South African education and training system, through its policy of outcomesbased education and training, has made competency a national priority. In compliance to this national requirement of producing competent learners, the South African Nursing Council ( 1999 B require that the beginner professional nurse practitioners and midwives have the necessary knowledge, skills, attitudes and values which will enable them to render efficient professional service. The health care system also demands competent nurse practitioners to ensure quality in health care. In the light of competency being a national priority and a statutory demand, the research question that emerges is, how competent are the newly qualified registered nurses from a specific nursing college in clinical nursing education? A quantitative, non-experimental contextual design was used to evaluate the competence of newly qualified registered nurses from a specific nursing college. The study was conducted in two phases. The first phase dealt with the development of an instrument together with its manual through the conceptualisation process. The second phase focused on the evaluation of the competency of newly qualified nurses using the instrument based on the steps of the nursing process. A pilot study was conducted to test the feasibility of the items of the instrument. During the evaluation phase, a sample of twenty-six newly qualified nurses was selected by simple random sampling from a target population of thirty-six newly qualified registered nurses. However, six participants withdrew from the study. Data was collected in two general hospitals where the newly qualified registered nurses were working. Observation and questioning were used as data collection techniques in accordance with the developed instrument. Measures were taken to ensure internal validity and reliability of the results. To protect the rights of the participants, the researcher adhered to DENOSA’S (1998

  7. Qualified expert training: the ERPC (European Radiation Protection Course)

    International Nuclear Information System (INIS)

    Deboodt, P.; Bourguignon, M.; Juhel, T.; Lirsac, N.; Luciani, A.; Marco, M.; Schmitt-Hanning, A. M.; Van der Steen, J.

    2003-01-01

    Since 2000, a training for the qualified experts is organised by several European partners in France at Saclay near Pais. The objective of this training is to deliver the theoretical knowledge needed to be recognised as a qualified expert in radiation protection according to the European requirements. The training delivered in English, includes four independent modules; basics, occupational exposure in nuclear and industrial applications, medical exposures (patients and workers), radiation protection of the members of the public and the environment (4 or 5 weeks each). A written examination is organised at the end of each module. Lectures, practical works, exercises and visits are given by European lecturers. Students have the possibility to register in one or several module during one or several years. Participants could be students or professionals from all European countries satisfying the pre requite defined by the European board. A certificate validating the successfully passed modules will be delivered by the INSTN (National Institute of Nuclear Sciences and Technologies)

  8. Simulation used to qualify nuclear waste glass for disposal

    International Nuclear Information System (INIS)

    Reimus, T.W.; Kuhn, W.L.

    1987-07-01

    A hypothetical vitrification system was simulated errors associated with controlling and predicting the composition of the nuclear waste glass produced in the system. The composition of the glass must fall within certain limits to qualify for permanent geologic disposal. The estimated error in predicting the concentrations of various constituents in the glass was 2% to 8%, depending on the strategy for sampling and analyzing the feed and on the assumed magnitudes of the process uncertainties. The estimated error in controlling the glass composition was 2% to 9%, depending on the strategy for sampling and analyzing the waste and on the assumed magnitudes of the uncertainties. This work demonstrates that simulation techniques can be used to assist in qualifying nuclear waste glass for disposal. 3 refs., 2 figs., 4 tabs

  9. Qualified expert training: the ERPC (European Radiation Protection Course)

    Energy Technology Data Exchange (ETDEWEB)

    Deboodt, P.; Bourguignon, M.; Juhel, T.; Lirsac, N.; Luciani, A.; Marco, M.; Schmitt-Hanning, A. M.; Van der Steen, J.

    2003-07-01

    Since 2000, a training for the qualified experts is organised by several European partners in France at Saclay near Pais. The objective of this training is to deliver the theoretical knowledge needed to be recognised as a qualified expert in radiation protection according to the European requirements. The training delivered in English, includes four independent modules; basics, occupational exposure in nuclear and industrial applications, medical exposures (patients and workers), radiation protection of the members of the public and the environment (4 or 5 weeks each). A written examination is organised at the end of each module. Lectures, practical works, exercises and visits are given by European lecturers. Students have the possibility to register in one or several module during one or several years. Participants could be students or professionals from all European countries satisfying the pre requite defined by the European board. A certificate validating the successfully passed modules will be delivered by the INSTN (National Institute of Nuclear Sciences and Technologies).

  10. Azithromycin effectiveness against intracellular infections of Francisella

    Directory of Open Access Journals (Sweden)

    Mann Barbara J

    2010-04-01

    Full Text Available Abstract Background Macrolide antibiotics are commonly administered for bacterial respiratory illnesses. Azithromycin (Az is especially noted for extremely high intracellular concentrations achieved within macrophages which is far greater than the serum concentration. Clinical strains of Type B Francisella (F. tularensis have been reported to be resistant to Az, however our laboratory Francisella strains were found to be sensitive. We hypothesized that different strains/species of Francisella (including Type A may have different susceptibilities to Az, a widely used and well-tolerated antibiotic. Results In vitro susceptibility testing of Az confirmed that F. tularensis subsp. holarctica Live Vaccine Strain (LVS (Type B was not sensitive while F. philomiragia, F. novicida, and Type A F. tularensis (NIH B38 and Schu S4 strain were susceptible. In J774A.1 mouse macrophage cells infected with F. philomiragia, F. novicida, and F. tularensis LVS, 5 μg/ml Az applied extracellularly eliminated intracellular Francisella infections. A concentration of 25 μg/ml Az was required for Francisella-infected A549 human lung epithelial cells, suggesting that macrophages are more effective at concentrating Az than epithelial cells. Mutants of RND efflux components (tolC and ftlC in F. novicida demonstrated less sensitivity to Az by MIC than the parental strain, but the tolC disc-inhibition assay demonstrated increased sensitivity, indicating a complex role for the outer-membrane transporter. Mutants of acrA and acrB mutants were less sensitive to Az than the parental strain, suggesting that AcrAB is not critical for the efflux of Az in F. novicida. In contrast, F. tularensis Schu S4 mutants ΔacrB and ΔacrA were more sensitive than the parental strain, indicating that the AcrAB may be important for Az efflux in F. tularensis Schu S4. F. novicida LPS O-antigen mutants (wbtN, wbtE, wbtQ and wbtA were found to be less sensitive in vitro to Az compared to the wild

  11. Gender Inequalities in Highly Qualified Professions: A Social Psychological Analysis

    OpenAIRE

    Santos, Maria Helena; Amâncio, Lígia

    2016-01-01

    Research in social and political psychology contributes towards understanding the persistence of job market gender segregation prevailing in recent decades, the consequences for those involved and their reactions when having to cope with gender inequality. Within the framework of the literature on shared ideologies that justify and legitimize discrimination against women, this article focuses on Portugal and analyses the particular case of women in two highly qualified professions traditional...

  12. Results from Consultancy Meeting on Qualified Technical Centres

    International Nuclear Information System (INIS)

    Novakovic, Mladen

    2017-01-01

    Qualified Technical Centres (QTC) - Current Situation: •Many Member States (MS) need help dealing with DSRS; •Most seek assistance, which can include funding, through the IAEA; •Technical assistance is provided by various governmental organization and private companies (service providers = SP). QTC - Method of Delivery Numerous MS requests are pressing the IAEA’s capacity to assist: •Use qualified internal and external experts/contractors; •Limited pool for DSRS and need to expand; •Dedicated and specialized equipment; •Mobile hot cell; •Mobile tool kit; •Field operations; •Coordinated approach with Nuclear Security and Safety. Qualified Technical Centres for the Management of DSRS: •The challenge is to scale up the support for safe and secure management of DSRS; •Strengthen existing capacity of some MS to support others and make support more sustainable; •Expands the support that can be readily provided and meet the many needs of MS

  13. Dietary habits of obese patients qualified for bariatric procedures.

    Science.gov (United States)

    Jastrzebska-Mierzyńska, Marta; Ostrowska, Lucyna; Hady, Hady Razak; Dadan, Jacek

    2014-01-01

    Patients with obesity, including morbid obesity, commit numerous dietary mistakes. They prefer high-energy diets, but of poor nutritional value. Patients qualified for bariatric procedures show deficiencies in vitamins and minerals due to insufficient intake of vegetables, fruit and whole grain products. Analysis of dietary habits in morbidly obese patients prepared for bariatric surgery, including assessment of eating style and frequency of consumption of certain products. The study group contained 39 people aged 18 - 65 years, who were surveyed with a questionnaire elaborated in the Department of Clinical Dietetics and Nutrition, Medical University of Bialystok. The following factors were assessed: number of meals, snacking between meals and eating at night, types of snacks eaten, and frequency of consumption of certain foods. Results were analyzed using Statistica 9.0. The majority of surveyed men (41.7%) ate three meals a day, whereas most women (40.7%) had at least 4 meals a day. Nearly 85% of the respondents admitted snacking between meals, mainly eating fruit, sweets and sandwiches. Whole grain cereal, milk and dairy products, fish, fruit, vegetables and pulses appeared in diet of patients qualified for treatment of obesity very rarely. Dietary habits of obese patients qualified for bariatric procedures are not consistent with recommendations. Therefore, these patients should receive nutritional education in order to foster proper eating habits that will help in the postoperative nutrition.

  14. Factor Structure of the Air Force Officer Qualifying Test: Analysis and Comparison

    National Research Council Canada - National Science Library

    Carreta, Thomas

    1998-01-01

    The Air Force Officer Qualifying Test (AFOQT) is used to qualify men and women for commissions in the Air Force, classify them for pilot and navigator jobs, and award Reserve Officer Training Corps (ROTC) scholarships...

  15. 47 CFR 59.4 - Definition of “qualifying carrier”.

    Science.gov (United States)

    2010-10-01

    ... (CONTINUED) INFRASTRUCTURE SHARING § 59.4 Definition of “qualifying carrier”. For purposes of this part, the term “qualifying carrier” means a telecommunications carrier that: (a) Lacks economies of scale or...

  16. Low-Income Housing Tax Credit (LIHTC) Qualified Census Tract (QCT)

    Data.gov (United States)

    Department of Housing and Urban Development — It allows to generate tables for Low-Income Housing Tax Credit (LIHTC) Qualified Census Tracts (QCT) and for Difficult Development Areas (DDA). LIHTC Qualified...

  17. Intracellular bacteria: the origin of dinoflagellate toxicity.

    Science.gov (United States)

    Silva, E S

    1990-01-01

    Dinoflagellate blooms of the same species have been registered either as toxic or nontoxic and, in the latter case, toxicity may be of different types. A hypothesis has been formulated according to which the bacteria having in some way taken part in the toxin formation are either inside the dinoflagellate cell or in the nutritive liquid. The presence of intracellular bacteria in those microorganisms has been studied mainly in material from cultures, a few from the sea, and several strains were isolated from different species. Experiments with crossed inoculations have shown that the bacterial strain from Gonyaulax tamarensis caused the cells of some other species to become toxic. From nontoxic clonal cultures of Prorocentrum balticum, Glenodinium foliaceum, and Gyrodinium instriatum, after inoculation of that bacterial strain, cultures were obtained whose cell extracts showed the same kind of toxicity as G. tamarensis. No toxic action could be found in the extracts of the bacterial cells form the assayed strains. The interference of intracellular bacteria in the metabolism of dinoflagellates must be the main cause of their toxicity.

  18. Fluorescent nanoparticles for intracellular sensing: a review.

    Science.gov (United States)

    Ruedas-Rama, Maria J; Walters, Jamie D; Orte, Angel; Hall, Elizabeth A H

    2012-11-02

    Fluorescent nanoparticles (NPs), including semiconductor NPs (Quantum Dots), metal NPs, silica NPs, polymer NPs, etc., have been a major focus of research and development during the past decade. The fluorescent nanoparticles show unique chemical and optical properties, such as brighter fluorescence, higher photostability and higher biocompatibility, compared to classical fluorescent organic dyes. Moreover, the nanoparticles can also act as multivalent scaffolds for the realization of supramolecular assemblies, since their high surface to volume ratio allow distinct spatial domains to be functionalized, which can provide a versatile synthetic platform for the implementation of different sensing schemes. Their excellent properties make them one of the most useful tools that chemistry has supplied to biomedical research, enabling the intracellular monitoring of many different species for medical and biological purposes. In this review, we focus on the developments and analytical applications of fluorescent nanoparticles in chemical and biological sensing within the intracellular environment. The review also points out the great potential of fluorescent NPs for fluorescence lifetime imaging microscopy (FLIM). Finally, we also give an overview of the current methods for delivering of fluorescent NPs into cells, where critically examine the benefits and liabilities of each strategy. Copyright © 2012 Elsevier B.V. All rights reserved.

  19. Mechanisms of cellular invasion by intracellular parasites.

    Science.gov (United States)

    Walker, Dawn M; Oghumu, Steve; Gupta, Gaurav; McGwire, Bradford S; Drew, Mark E; Satoskar, Abhay R

    2014-04-01

    Numerous disease-causing parasites must invade host cells in order to prosper. Collectively, such pathogens are responsible for a staggering amount of human sickness and death throughout the world. Leishmaniasis, Chagas disease, toxoplasmosis, and malaria are neglected diseases and therefore are linked to socio-economical and geographical factors, affecting well-over half the world's population. Such obligate intracellular parasites have co-evolved with humans to establish a complexity of specific molecular parasite-host cell interactions, forming the basis of the parasite's cellular tropism. They make use of such interactions to invade host cells as a means to migrate through various tissues, to evade the host immune system, and to undergo intracellular replication. These cellular migration and invasion events are absolutely essential for the completion of the lifecycles of these parasites and lead to their for disease pathogenesis. This review is an overview of the molecular mechanisms of protozoan parasite invasion of host cells and discussion of therapeutic strategies, which could be developed by targeting these invasion pathways. Specifically, we focus on four species of protozoan parasites Leishmania, Trypanosoma cruzi, Plasmodium, and Toxoplasma, which are responsible for significant morbidity and mortality.

  20. [Intracellular signaling mechanisms in thyroid cancer].

    Science.gov (United States)

    Mondragón-Terán, Paul; López-Hernández, Luz Berenice; Gutiérrez-Salinas, José; Suárez-Cuenca, Juan Antonio; Luna-Ceballos, Rosa Isela; Erazo Valle-Solís, Aura

    2016-01-01

    Thyroid cancer is the most common malignancy of the endocrine system, the papillary variant accounts for 80-90% of all diagnosed cases. In the development of papillary thyroid cancer, BRAF and RAS genes are mainly affected, resulting in a modification of the system of intracellular signaling proteins known as «protein kinase mitogen-activated» (MAPK) which consist of «modules» of internal signaling proteins (Receptor/Ras/Raf/MEK/ERK) from the cell membrane to the nucleus. In thyroid cancer, these signanling proteins regulate diverse cellular processes such as differentiation, growth, development and apoptosis. MAPK play an important role in the pathogenesis of thyroid cancer as they are used as molecular biomarkers for diagnostic, prognostic and as possible therapeutic molecular targets. Mutations in BRAF gene have been correlated with poor response to treatment with traditional chemotherapy and as an indicator of poor prognosis. To review the molecular mechanisms involved in intracellular signaling of BRAF and RAS genes in thyroid cancer. Molecular therapy research is in progress for this type of cancer as new molecules have been developed in order to inhibit any of the components of the signaling pathway (RET/PTC)/Ras/Raf/MEK/ERK; with special emphasis on the (RET/PTC)/Ras/Raf section, which is a major effector of ERK pathway. Copyright © 2016 Academia Mexicana de Cirugía A.C. Publicado por Masson Doyma México S.A. All rights reserved.

  1. The Netherlands: Quality of work and employment of low-qualified workers

    NARCIS (Netherlands)

    Gouw, P.

    2013-01-01

    Based on the Netherlands Working Condition Survey (NEA, TNO) and the available literature, it seems that low-qualified workers experience more labour market problems than middle- or high-qualified workers. The trend in the 2000-2007 period is that the relative number of low qualified workers has

  2. 29 CFR 779.386 - Restaurants may qualify as exempt 13(a)(2) establishments.

    Science.gov (United States)

    2010-07-01

    ... 29 Labor 3 2010-07-01 2010-07-01 false Restaurants may qualify as exempt 13(a)(2) establishments... Service Establishments Restaurants and Establishments Providing Food and Beverage Service § 779.386 Restaurants may qualify as exempt 13(a)(2) establishments. (a) A restaurant may qualify as an exempt retail or...

  3. 6 CFR 25.4 - Designation of qualified anti-terrorism technologies.

    Science.gov (United States)

    2010-01-01

    ... 6 Domestic Security 1 2010-01-01 2010-01-01 false Designation of qualified anti-terrorism... REGULATIONS TO SUPPORT ANTI-TERRORISM BY FOSTERING EFFECTIVE TECHNOLOGIES § 25.4 Designation of qualified anti-terrorism technologies. (a) General. The Under Secretary may Designate as a Qualified Anti-Terrorism...

  4. 12 CFR 619.9270 - Qualified Public Accountant or External Auditor.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 6 2010-01-01 2010-01-01 false Qualified Public Accountant or External Auditor... § 619.9270 Qualified Public Accountant or External Auditor. A qualified public accountant or external... constituted State authority, identifying such person as a certified public accountant; (b) Is licensed to...

  5. 49 CFR 192.285 - Plastic pipe: Qualifying persons to make joints.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 3 2010-10-01 2010-10-01 false Plastic pipe: Qualifying persons to make joints... Materials Other Than by Welding § 192.285 Plastic pipe: Qualifying persons to make joints. (a) No person may make a plastic pipe joint unless that person has been qualified under the applicable joining procedure...

  6. 10 CFR 451.4 - What is a qualified renewable energy facility.

    Science.gov (United States)

    2010-01-01

    ... 10 Energy 3 2010-01-01 2010-01-01 false What is a qualified renewable energy facility. 451.4 Section 451.4 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION RENEWABLE ENERGY PRODUCTION INCENTIVES § 451.4 What is a qualified renewable energy facility. In order to qualify for an incentive payment under...

  7. 26 CFR 1.468B-2 - Taxation of qualified settlement funds and related administrative requirements.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 6 2010-04-01 2010-04-01 false Taxation of qualified settlement funds and... Taken § 1.468B-2 Taxation of qualified settlement funds and related administrative requirements. (a) In...) of this section is in lieu of any other taxation of the income of a qualified settlement fund under...

  8. Nanobodies: Chemical Functionalization Strategies and Intracellular Applications

    Science.gov (United States)

    Schumacher, Dominik; Helma, Jonas; Schneider, Anselm F. L.; Leonhardt, Heinrich

    2018-01-01

    Abstract Nanobodies can be seen as next‐generation tools for the recognition and modulation of antigens that are inaccessible to conventional antibodies. Due to their compact structure and high stability, nanobodies see frequent usage in basic research, and their chemical functionalization opens the way towards promising diagnostic and therapeutic applications. In this Review, central aspects of nanobody functionalization are presented, together with selected applications. While early conjugation strategies relied on the random modification of natural amino acids, more recent studies have focused on the site‐specific attachment of functional moieties. Such techniques include chemoenzymatic approaches, expressed protein ligation, and amber suppression in combination with bioorthogonal modification strategies. Recent applications range from sophisticated imaging and mass spectrometry to the delivery of nanobodies into living cells for the visualization and manipulation of intracellular antigens. PMID:28913971

  9. Drosophila VAMP7 regulates Wingless intracellular trafficking.

    Science.gov (United States)

    Gao, Han; He, Fang; Lin, Xinhua; Wu, Yihui

    2017-01-01

    Drosophila Wingless (Wg) is a morphogen that determines cell fate during development. Previous studies have shown that endocytic pathways regulate Wg trafficking and signaling. Here, we showed that loss of vamp7, a gene required for vesicle fusion, dramatically increased Wg levels and decreased Wg signaling. Interestingly, we found that levels of Dally-like (Dlp), a glypican that can interact with Wg to suppress Wg signaling at the dorsoventral boundary of the Drosophila wing, were also increased in vamp7 mutant cells. Moreover, Wg puncta in Rab4-dependent recycling endosomes were Dlp positive. We hypothesize that VAMP7 is required for Wg intracellular trafficking and the accumulation of Wg in Rab4-dependent recycling endosomes might affect Wg signaling.

  10. Intracellular pH in sperm physiology.

    Science.gov (United States)

    Nishigaki, Takuya; José, Omar; González-Cota, Ana Laura; Romero, Francisco; Treviño, Claudia L; Darszon, Alberto

    2014-08-01

    Intracellular pH (pHi) regulation is essential for cell function. Notably, several unique sperm ion transporters and enzymes whose elimination causes infertility are either pHi dependent or somehow related to pHi regulation. Amongst them are: CatSper, a Ca(2+) channel; Slo3, a K(+) channel; the sperm-specific Na(+)/H(+) exchanger and the soluble adenylyl cyclase. It is thus clear that pHi regulation is of the utmost importance for sperm physiology. This review briefly summarizes the key components involved in pHi regulation, their characteristics and participation in fundamental sperm functions such as motility, maturation and the acrosome reaction. Copyright © 2014 Elsevier Inc. All rights reserved.

  11. Nanobodies: Chemical Functionalization Strategies and Intracellular Applications.

    Science.gov (United States)

    Schumacher, Dominik; Helma, Jonas; Schneider, Anselm F L; Leonhardt, Heinrich; Hackenberger, Christian P R

    2018-02-23

    Nanobodies can be seen as next-generation tools for the recognition and modulation of antigens that are inaccessible to conventional antibodies. Due to their compact structure and high stability, nanobodies see frequent usage in basic research, and their chemical functionalization opens the way towards promising diagnostic and therapeutic applications. In this Review, central aspects of nanobody functionalization are presented, together with selected applications. While early conjugation strategies relied on the random modification of natural amino acids, more recent studies have focused on the site-specific attachment of functional moieties. Such techniques include chemoenzymatic approaches, expressed protein ligation, and amber suppression in combination with bioorthogonal modification strategies. Recent applications range from sophisticated imaging and mass spectrometry to the delivery of nanobodies into living cells for the visualization and manipulation of intracellular antigens. © 2017 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

  12. Intracellular Signalling by C-Peptide

    Directory of Open Access Journals (Sweden)

    Claire E. Hills

    2008-01-01

    Full Text Available C-peptide, a cleavage product of the proinsulin molecule, has long been regarded as biologically inert, serving merely as a surrogate marker for insulin release. Recent findings demonstrate both a physiological and protective role of C-peptide when administered to individuals with type I diabetes. Data indicate that C-peptide appears to bind in nanomolar concentrations to a cell surface receptor which is most likely to be G-protein coupled. Binding of C-peptide initiates multiple cellular effects, evoking a rise in intracellular calcium, increased PI-3-kinase activity, stimulation of the Na+/K+ ATPase, increased eNOS transcription, and activation of the MAPK signalling pathway. These cell signalling effects have been studied in multiple cell types from multiple tissues. Overall these observations raise the possibility that C-peptide may serve as a potential therapeutic agent for the treatment or prevention of long-term complications associated with diabetes.

  13. Intracellular sphingosine releases calcium from lysosomes.

    Science.gov (United States)

    Höglinger, Doris; Haberkant, Per; Aguilera-Romero, Auxiliadora; Riezman, Howard; Porter, Forbes D; Platt, Frances M; Galione, Antony; Schultz, Carsten

    2015-11-27

    To elucidate new functions of sphingosine (Sph), we demonstrate that the spontaneous elevation of intracellular Sph levels via caged Sph leads to a significant and transient calcium release from acidic stores that is independent of sphingosine 1-phosphate, extracellular and ER calcium levels. This photo-induced Sph-driven calcium release requires the two-pore channel 1 (TPC1) residing on endosomes and lysosomes. Further, uncaging of Sph leads to the translocation of the autophagy-relevant transcription factor EB (TFEB) to the nucleus specifically after lysosomal calcium release. We confirm that Sph accumulates in late endosomes and lysosomes of cells derived from Niemann-Pick disease type C (NPC) patients and demonstrate a greatly reduced calcium release upon Sph uncaging. We conclude that sphingosine is a positive regulator of calcium release from acidic stores and that understanding the interplay between Sph homeostasis, calcium signaling and autophagy will be crucial in developing new therapies for lipid storage disorders such as NPC.

  14. [Limbic encephalitis with antibodies against intracellular antigens].

    Science.gov (United States)

    Morita, Akihiko; Kamei, Satoshi

    2010-04-01

    Limbic encephalitis is a paraneoplastic syndrome that is often associated with small cell lung cancer (SCLC), breast cancer, testicular tumors, teratoma, Hodgkin's lymphoma and thymoma. The common clinical manifestations of limbic encephalitis are subacute onset, cognitive dysfunction, seizures and psychiatric symptoms. Paraneoplastic neurological disorders are considered to occur because of cytotoxic T cell responses and antibodies against target neuronal proteins that are usually expressed by an underlying tumor. The main intracellular antigens related to limbic encephalitis are Hu, Ma2, and less frequently CV2/CRMP5 and amphiphysin. The anti-Hu antibody, which is involved in cerebellar degeneration and extensive or multifocal encephalomyelitis such as limbic encephalitis is closely associated with a history of smoking and SCLC. The anti-Ma2 antibody is associated with encephalitis of the limbic system, hypothalamus and brain-stem. For this reason, some patients with limbic encephalitis have sleep disorders (including REM sleep abnormalities), severe hypokinesis and gaze palsy in addition to limbic dysfunction. In men aged less than 50 years, anti-Ma2 antibody encephalitis is almost always associated with testicular germ-cell tumors that are occasionally difficult to detect. In older men and women, the most common tumors are non-SCLC and breast cancer. Limbic encephalitis associated with cell-surface antigens (e.g., voltage-gated potassium channels, NMDA receptors) is mediated by antibodies and often improves after a reduction in the antibody titer and after tumor resection. Patients with antibodies against intracellular antigens, except for those with anti-Ma2 antibodies and testicular tumors, are less responsive. Early diagnosis and treatment with immunotherapy, tumor resection or both are important for improving or stabilizing the condition of limbic encephalitis.

  15. Registration of qualified radiation protection experts in the Netherlands

    International Nuclear Information System (INIS)

    Abrahamse, J.C.; Kops, J.A.M.M.; Vermeulen, A.M.T.I.; Zuur, C.

    2002-01-01

    In Euratom Directive 96/29 (1) the Euratom Basic Safety Standards are given. The member states of the EC have to implement this directive in their national legislation. In the Netherlands this is done in the new radiation protection legislation (Radiation Protection Decree 2002 (2)), which came into force in the first half of 2002. In this new regulation is stated that radiological activities must be done by or under supervision of a qualified radiation protection expert. With respect to the qualified expert is stated that: the qualified expert must be registered in a notified registration system; rules will be formulated with respect to the demanded qualification levels of the experts. These demands can be different for different tasks rules will be formulated for the notification of the register, the way to become registered, the costs and so on. Mid 2001 a project is started to prepare the registration system. Information was gained from and discussions took place with the important parties on the radiological playing field in the Netherlands (employers, organisations of experts and education institutes). Furthermore information is obtained from certification institutes. It was decided to accept as starting point the education levels obtained from the radiation protection education institutes, which are approved at the moment. At a later phase will be evaluated whether this system needs to be changed. In section 2 of this paper the radiation protection system in the Netherlands will be discussed in more detail. Section 2 deals with the target group and in section 3 the outlines of the registration system to be developed are discussed

  16. Professional paradox: identity formation in qualified doctors pursuing further training.

    Science.gov (United States)

    Chan, Mercedes; Pratt, Dan; Poole, Gary; Sidhu, Ravi

    2018-03-01

    Many newly qualified specialists and subspecialists pursue additional training. Although their motivations are many, the pursuit of further training as an alternative to unemployment is an emerging trend. Paradoxically, doctors continue as trainees with a consultant's credentials, and without the guarantee of eventual employment. This study explores seven doctors' experiences, the effects of further training on their professional identity formation (PIF), and how these effects are reconciled on a personal and professional level. This phenomenological study involved interviews with seven qualified Canadian specialists (three were female) who pursued additional training in response to a lack of available positions in their respective specialties. Template analysis generated theoretical constructs of influences on their PIF, and characteristics of their lived experiences. Four themes shaped PIF: setting and context; language and communication; responsibilities and privileges; and participants' visions of their future selves. Professional identity formation (PIF) continued to develop in further training, but was inconsistently affirmed by participants' communities of practice. Four major themes characterised training experiences: prescription; managing multiple masters; limiting access to others and community ties; and constantly questioning the value of extra training. Qualified doctors traverse professional paradoxes as they seek further education with no guarantee of employment and provide consultant-level care as 'trainees'. An identity dissonance emerges that may continue until a clear identity is prescribed for them. Although disruptive to these doctors' PIF and personal and professional lives, the long-term effects of additional training are unknown. Its utility and influence on securing employment and future job satisfaction are areas for further research. © 2017 John Wiley & Sons Ltd and The Association for the Study of Medical Education.

  17. Qualifying works of the Department of nuclear chemistry (1963 - 2006)

    International Nuclear Information System (INIS)

    Kuruc, J.

    2007-01-01

    In this review qualifying works (theses - bachelor, master, PhD., DrSc., habilitation and inauguration theses) elaborated at the Department of nuclear chemistry, Faculty of Natural Chemistry, Comenius University in Bratislava during forty years (from origin of the Section of Nuclear chemistry in 1963 up to 2006 are presented. During this time, in totally, 3 bachelor theses, 265 master theses, 24 PhD. (CSc.) and 10 PhD. dissertanions, 2 DrSc. dissertanions as well as 8 habilitation and one inauguration these were defended (author)

  18. Qualified equipment for spare parts and modifications in Belgium

    International Nuclear Information System (INIS)

    Berthe, J.

    1993-01-01

    This paper describes the procedure followed for the procurement of components for spare parts and systems modifications, the legal and quality assurance environment and the specific measures taken to cope with various codes and standards. For pressure components, the regulation aspects of the American code used during the construction phase of plants had to be adapted to the Belgian context and to the actual industry situation. Obsolescence of products is treated either by qualification of a new product or by design modifications, or by replacement with an already qualified product. In these cases, the work is handled by the utility's engineering organization. 2 figs

  19. Hiring and keeping qualified personnel in a growing regulatory organisation

    International Nuclear Information System (INIS)

    Marchildon, P.

    1993-01-01

    Although no new nuclear stations are currently committed in Canada, the regulatory authority, the Atomic Energy Control Board (AECB), has started, in 1989, an expansion program which, when completed in 1994 will represent a 60 pc increase in its workforce. The new personnel resources are being used to improve the AECB effectiveness in a number of regulatory activities. This paper reviews some of the key factors which relate to the hiring and integration of the new AECB employees: selection process, timing, fees, scholarship program, subsequent training of qualified personnel, woman quotas. 2 figs

  20. Bank Officers’ Perceptions and Uses of Qualified Audit Reports

    DEFF Research Database (Denmark)

    Sormunen, Nina

    2014-01-01

    Purpose – The purpose of this study is to provide insights into the perceptions and uses of qualified audit reports in financial statements of small- and medium-sized enterprises (SMEs). As there is a long-standing debate on the usefulness of auditor’s going-concern reports, this study aims....... Practical implications – Regulators have taken the action to improve the auditor’s reporting model, and the findings provided by this study are important because they provide a deeper understanding of the perceptions and uses of audit reports from smaller companies. The study also contributes knowledge...

  1. Single-cell intracellular nano-pH probes†

    OpenAIRE

    Özel, Rıfat Emrah; Lohith, Akshar; Mak, Wai Han; Pourmand, Nader

    2015-01-01

    Within a large clonal population, such as cancerous tumor entities, cells are not identical, and the differences between intracellular pH levels of individual cells may be important indicators of heterogeneity that could be relevant in clinical practice, especially in personalized medicine. Therefore, the detection of the intracellular pH at the single-cell level is of great importance to identify and study outlier cells. However, quantitative and real-time measurements of the intracellular p...

  2. 30 CFR 203.43 - To which production do I apply the RSV earned from qualified deep wells or qualified phase 1...

    Science.gov (United States)

    2010-07-01

    ... in water between 200 and 400 meters deep, you begin drilling an original deep well with a perforated... 200 meters deep; (ii) May 18, 2007, for an RSV earned by a qualified deep well on a lease that is located entirely in water more than 200 meters deep; or (iii) The date that the first qualified well that...

  3. Statistically qualified neuro-analytic failure detection method and system

    Science.gov (United States)

    Vilim, Richard B.; Garcia, Humberto E.; Chen, Frederick W.

    2002-03-02

    An apparatus and method for monitoring a process involve development and application of a statistically qualified neuro-analytic (SQNA) model to accurately and reliably identify process change. The development of the SQNA model is accomplished in two stages: deterministic model adaption and stochastic model modification of the deterministic model adaptation. Deterministic model adaption involves formulating an analytic model of the process representing known process characteristics, augmenting the analytic model with a neural network that captures unknown process characteristics, and training the resulting neuro-analytic model by adjusting the neural network weights according to a unique scaled equation error minimization technique. Stochastic model modification involves qualifying any remaining uncertainty in the trained neuro-analytic model by formulating a likelihood function, given an error propagation equation, for computing the probability that the neuro-analytic model generates measured process output. Preferably, the developed SQNA model is validated using known sequential probability ratio tests and applied to the process as an on-line monitoring system. Illustrative of the method and apparatus, the method is applied to a peristaltic pump system.

  4. Spatial factors as contextual qualifiers of information seeking

    Directory of Open Access Journals (Sweden)

    R. Savolainen

    2006-01-01

    Full Text Available Introduction. This paper investigates the ways in which spatial factors have been approached in information seeking studies. The main attention was focused on studies discussing information seeking on the level of source selection and use. Method. Conceptual analysis of about 100 articles and books thematizing spatial issues of information seeking. Due to research economy, the main attention was paid to studies on everyday life information seeking. Results. Three major viewpoints were identified with regard to the degree of objectivity of spatial factors. The objectifying approach conceives of spatial factors as external and entity-like qualifiers that primarly constrain information seeking. The realistic-pragmatic approach emphasizes the ways in which the availabilty of information sources in different places such as daily work environments orient information seeking. The perspectivist approach focuses on how people subjectively assess the significance of various sources by means of spatial constructs such as information horizons. Conclusion. Spatial factors are centrally important contextual qualifiers of information seeking. There is a need to further explore the potential of the above viewpoints by relating the spatial and temporal factors of information seeking.

  5. Strategy for development modes competent performance in teachers qualified alternative

    Directory of Open Access Journals (Sweden)

    Sailí Rodríguez Fuentes

    2017-12-01

    Full Text Available Introduction: the process of professionalization of the teacher is a subject of contemporary conjuncture, which is debated and investigated by researchers from dissimilar edges. Objective: to contribute to the professionalization process of qualified teachers of Technical and Vocational Education through a pedagogical strategy. Materials and methods: The materialist dialectic is used as a general method, allowing the study of the object as a process, the determination of its components, as well as its contradictions. theoretical methods: the historical and logical to delve into the regularities and qualities of reality in the polytechnic centers. Essential empirical methods were used: the documentary analysis that included the professional model; study plans, to obtain authentic and reliable information for research, interviews with students with the purpose of analyzing the manifestations of the professionalization process of the students and the direction of the process by the teachers, the observation of teaching activities allowed to diagnose the object , the demonstration of the problem, as well as the results of the practice of the strategy to be implemented. Results: a pedagogical strategy is designed and implemented to perfect the competent professional performance modes in the qualified professors. Conclusions: it contributed to the improvement of the professionalization process in favor of the competent professional modes of action.

  6. LIPID SYNTHESIS, INTRACELLULAR TRANSPORT, AND SECRETION

    Science.gov (United States)

    Stein, Olga; Stein, Yechezkiel

    1967-01-01

    In the mammary glands of lactating albino mice injected intravenously with 9, 10-oleic acid-3H or 9, 10-palmitic acid-3H, it has been shown that the labeled fatty acids are incorporated into mammary gland glycerides. The labeled lipid in the mammary gland 1 min after injection was in esterified form (> 95%), and the radioautographic reaction was seen over the rough endoplasmic reticulum and over lipid droplets, both intracellular and intraluminal. At 10–60 min after injection, the silver grains were concentrated predominantly over lipid droplets. There was no concentration of radioactivity over the granules in the Golgi apparatus, at any time interval studied. These findings were interpreted to indicate that after esterification of the fatty acid into glycerides in the rough endoplasmic reticulum an in situ aggregation of lipid occurs, with acquisition of droplet form. The release of the lipid into the lumen proceeds directly and not through the Golgi apparatus, in contradistinction to the mode of secretion of casein in the mammary gland or of lipoprotein in the liver. The presence of strands of endoplasmic reticulum attached to intraluminal lipid droplets provides a structural counterpart to the milk microsomes described in ruminant milk. PMID:6033535

  7. Endothelial remodelling and intracellular calcium machinery.

    Science.gov (United States)

    Moccia, F; Tanzi, F; Munaron, L

    2014-05-01

    Rather being an inert barrier between vessel lumen and surrounding tissues, vascular endothelium plays a key role in the maintenance of cardiovascular homeostasis. The de-endothelialization of blood vessels is regarded as the early event that results in the onset of severe vascular disorders, including atherosclerosis, acute myocardial infarction, brain stroke, and aortic aneurysm. Restoration of the endothelial lining may be accomplished by the activation of neighbouring endothelial cells (ECs) freed by contact inhibition and by circulating endothelial progenitor cells (EPCs). Intracellular Ca(2+) signalling is essential to promote wound healing: however, the molecular underpinnings of the Ca(2+) response to injury are yet to be fully elucidated. Similarly, the components of the Ca(2+) toolkit that drive EPC incorporation into denuded vessels are far from being fully elucidated. The present review will survey the current knowledge on the role of Ca(2+) signalling in endothelial repair and in EPC activation. We propose that endothelial regeneration might be boosted by intraluminal release of specific Ca(2+) channel agonists or by gene transfer strategies aiming to enhance the expression of the most suitable Ca(2+) channels at the wound site. In this view, connexin (Cx) channels/hemichannels and store-operated Ca(2+) entry (SOCE) stand amid the most proper routes to therapeutically induce the regrowth of denuded vessels. Cx stimulation might trigger the proliferative and migratory behaviour of ECs facing the lesion site, whereas activation of SOCE is likely to favour EPC homing to the wounded vessel.

  8. Intracellular events regulating cross-presentation

    Directory of Open Access Journals (Sweden)

    Peter eCresswell

    2012-06-01

    Full Text Available Cross-presentation plays a fundamental role in the induction of CD8-T cell immunity. However, although more than three decades have passed since its discovery, surprisingly little is known about the exact mechanisms involved. Here we give an overview of the components involved at different stages of this process. First, antigens must be internalized into the cross-presenting cell. The involvement of different receptors, method of antigen uptake, and nature of the antigen can influence intracellular trafficking and access to the cross-presentation pathway. Once antigens access the endocytic system, different requirements for endosomal/phagosomal processing arise, such as proteolysis and reduction of disulfide bonds. The majority of cross-presented peptides are generated by proteasomal degradation. Therefore, antigens must cross a membrane barrier in a manner analogous to the fate of misfolded proteins in the endoplasmic reticulum (ER that are retrotranslocated into the cytosol for degradation. Indeed, some components of the ER-associated degradation (ERAD machinery have been implicated in cross-presentation. Further complicating the matter, endosomal and phagosomal compartments have been suggested as alternative sites to the ER for loading of peptides on MHC class I molecules. Finally, the antigen presenting cells involved, particularly dendritic cell subsets and their state of maturation, influence the efficiency of cross-presentation.

  9. Intracellular recording from a spider vibration receptor.

    Science.gov (United States)

    Gingl, Ewald; Burger, Anna-M; Barth, Friedrich G

    2006-05-01

    The present study introduces a new preparation of a spider vibration receptor that allows intracellular recording of responses to natural mechanical or electrical stimulation of the associated mechanoreceptor cells. The spider vibration receptor is a lyriform slit sense organ made up of 21 cuticular slits located on the distal end of the metatarsus of each walking leg. The organ is stimulated when the tarsus receives substrate vibrations, which it transmits to the organ's cuticular structures, reducing the displacement to about one tenth due to geometrical reasons. Current clamp recording was used to record action potentials generated by electrical or mechanical stimuli. Square pulse stimulation identified two groups of sensory cells, the first being single-spike cells which generated only one or two action potentials and the second being multi-spike cells which produced bursts of action potentials. When the more natural mechanical sinusoidal stimulation was applied, differences in adaptation rate between the two cell types remained. In agreement with prior extracellular recordings, both cell types showed a decrease in the threshold tarsus deflection with increasing stimulus frequency. Off-responses to mechanical stimuli have also been seen in the metatarsal organ for the first time.

  10. On the Computing Potential of Intracellular Vesicles.

    Science.gov (United States)

    Mayne, Richard; Adamatzky, Andrew

    2015-01-01

    Collision-based computing (CBC) is a form of unconventional computing in which travelling localisations represent data and conditional routing of signals determines the output state; collisions between localisations represent logical operations. We investigated patterns of Ca2+-containing vesicle distribution within a live organism, slime mould Physarum polycephalum, with confocal microscopy and observed them colliding regularly. Vesicles travel down cytoskeletal 'circuitry' and their collisions may result in reflection, fusion or annihilation. We demonstrate through experimental observations that naturally-occurring vesicle dynamics may be characterised as a computationally-universal set of Boolean logical operations and present a 'vesicle modification' of the archetypal CBC 'billiard ball model' of computation. We proceed to discuss the viability of intracellular vesicles as an unconventional computing substrate in which we delineate practical considerations for reliable vesicle 'programming' in both in vivo and in vitro vesicle computing architectures and present optimised designs for both single logical gates and combinatorial logic circuits based on cytoskeletal network conformations. The results presented here demonstrate the first characterisation of intracelluar phenomena as collision-based computing and hence the viability of biological substrates for computing.

  11. Modeling HIV-1 intracellular replication: two simulation approaches

    NARCIS (Netherlands)

    Zarrabi, N.; Mancini, E.; Tay, J.; Shahand, S.; Sloot, P.M.A.

    2010-01-01

    Many mathematical and computational models have been developed to investigate the complexity of HIV dynamics, immune response and drug therapy. However, there are not many models which consider the dynamics of virus intracellular replication at a single level. We propose a model of HIV intracellular

  12. Intracellular Drug Bioavailability: Effect of Neutral Lipids and Phospholipids.

    Science.gov (United States)

    Treyer, Andrea; Mateus, André; Wiśniewski, Jacek R; Boriss, Hinnerk; Matsson, Pär; Artursson, Per

    2018-06-04

    Intracellular unbound drug concentrations are the pharmacologically relevant concentrations for targets inside cells. Intracellular drug concentrations are determined by multiple processes, including the extent of drug binding to intracellular structures. The aim of this study was to evaluate the effect of neutral lipid (NL) and phospholipid (PL) levels on intracellular drug disposition. The NL and/or PL content of 3T3-L1 cells were enhanced, resulting in phenotypes (in terms of morphology and proteome) reminiscent of adipocytes (high NL and PL) or mild phospholipidosis (only high PL). Intracellular bioavailability ( F ic ) was then determined for 23 drugs in these cellular models and in untreated wild-type cells. A higher PL content led to higher intracellular drug binding and a lower F ic . The induction of NL did not further increase drug binding but led to altered F ic due to increased lysosomal pH. Further, there was a good correlation between binding to beads coated with pure PL and intracellular drug binding. In conclusion, our results suggest that PL content is a major determinant of drug binding in cells and that PL beads may constitute a simple alternative to estimating this parameter. Further, the presence of massive amounts of intracellular NLs did not influence drug binding significantly.

  13. Analysis of Intracellular Metabolites from Microorganisms: Quenching and Extraction Protocols.

    Science.gov (United States)

    Pinu, Farhana R; Villas-Boas, Silas G; Aggio, Raphael

    2017-10-23

    Sample preparation is one of the most important steps in metabolome analysis. The challenges of determining microbial metabolome have been well discussed within the research community and many improvements have already been achieved in last decade. The analysis of intracellular metabolites is particularly challenging. Environmental perturbations may considerably affect microbial metabolism, which results in intracellular metabolites being rapidly degraded or metabolized by enzymatic reactions. Therefore, quenching or the complete stop of cell metabolism is a pre-requisite for accurate intracellular metabolite analysis. After quenching, metabolites need to be extracted from the intracellular compartment. The choice of the most suitable metabolite extraction method/s is another crucial step. The literature indicates that specific classes of metabolites are better extracted by different extraction protocols. In this review, we discuss the technical aspects and advancements of quenching and extraction of intracellular metabolite analysis from microbial cells.

  14. Analysis of Intracellular Metabolites from Microorganisms: Quenching and Extraction Protocols

    Directory of Open Access Journals (Sweden)

    Farhana R. Pinu

    2017-10-01

    Full Text Available Sample preparation is one of the most important steps in metabolome analysis. The challenges of determining microbial metabolome have been well discussed within the research community and many improvements have already been achieved in last decade. The analysis of intracellular metabolites is particularly challenging. Environmental perturbations may considerably affect microbial metabolism, which results in intracellular metabolites being rapidly degraded or metabolized by enzymatic reactions. Therefore, quenching or the complete stop of cell metabolism is a pre-requisite for accurate intracellular metabolite analysis. After quenching, metabolites need to be extracted from the intracellular compartment. The choice of the most suitable metabolite extraction method/s is another crucial step. The literature indicates that specific classes of metabolites are better extracted by different extraction protocols. In this review, we discuss the technical aspects and advancements of quenching and extraction of intracellular metabolite analysis from microbial cells.

  15. Dimensions of taste qualifying didactic reflections on home economics education

    DEFF Research Database (Denmark)

    Wistoft, Karen; Qvortrup, Lars

    Aim: Traditionally and worldwide home economics education has been concerned with taste and flavorings based training associated with students cooking in the school kitchen. In contrast new educational research on food knowledge in the Danish public school shows that taste is used as a didactic...... review on children, learning, food and taste followed by analyses in a value reflective pedagogy perspective as well as quantitative and qualitative research in home economic education. This has been combined with systems theory developed by the German sociologist Niklas Luhmann providing concepts...... element and "overtakes" more traditional didactic elements like motivation and active participation. The purpose of this proposal is to present a systematic model for qualifying reflections on taste in home economics and food education. The objective is to identify four dimensions of taste that can...

  16. Information Support of Highly-Qualified Archers’ Training

    Directory of Open Access Journals (Sweden)

    А. П. Власов

    2017-03-01

    Full Text Available The objective is to ground the ways of improving the level of technical preparedness of highly-qualified archers by means of video and multimedia technologies. Materials and methods: The research studied motor actions of 25 athletes of the national team of Ukraine in archery when they were performing training exercises indoors (exercising area and outdoors (archery range. Each athlete’s technique of executing shooting exercises was video recorded in sequence from different angles (from above, from behind, from the front, from the left and the right sides depending on the shooting line with a digital video camera Sony DCR-XR150E. The processing of the video materials was done with the Dartfish Connekt software package. Results: The research grounds the need to use video and multimedia technologies in the process of training of highly-qualified archers. Such methods aim at improving the skills of both coaches and athletes by helping them better understand all aspects of the training exercises execution. Multimedia means offer a delayed-in-time opportunity to watch the video recording of the training after its preliminary computer processing. It enables the coach and the athlete to focus on the analysis of the execution of the entire exercise and its elements. The paper provides examples of the use of video technologies in archers’ training practice. Conclusions: Prospects of the use of the state-of-the-art computer technologies in sports training are infinite. Computer software and hardware systems should be available for most coaches. The task of the latter is to learn to use and implement them for the benefit of themselves and athletes. The task of the scientific groups and experts is to find a most convenient way for the coach to use the innovative technologies. The task of the software developer is to model intellectual programs to help the coach effectively analyze and evaluate his archers or team when preparing them for the most

  17. Attitude to e-learning among newly qualified doctors.

    Science.gov (United States)

    Goh, Jason; Clapham, Michael

    2014-02-01

    e-Learning plays an increasingly important role in medical education. Much research has focused on the evaluation of individual modules among medical students or more senior trainee doctors. We studied the attitude of newly qualified foundation level-1 doctors (FY1s) towards a blended learning programme to gain insight into the perceived role of e-learning in relation to classroom and experiential learning. The blended learning strategy consisted of weekly 3-hour sessions of lectures and flexible e-learning sessions. A questionnaire survey was conducted among 54 per cent (37/69) of FY1 doctors, towards the end of their first year post qualification. The majority of FY1s had to carry out additional e-learning outside of work. When asked where was best to carry out e-learning, 54 per cent preferred to e-learn both at work and at home, whereas 38 per cent preferred to e-learn outside of work exclusively. An equal preference for a classroom-only strategy and a blended programme was reported. Seventy-three per cent of the FY1s thought that e-learning should not be part of their compulsory weekly teaching programme. Fifty-four per cent of FY1s thought that e-learning had been useful for their education and training in their FY1 year. The e-learning package cited as being most useful was the safe prescribing e-programme, pioneered locally. Newly qualified doctors value e-learning as an adjunct to experiential and lecture-based teaching, and most prefer it as part of a blended learning programme at work or at home. Medical educators must place equal emphasis on the delivery and administration of e-learning as well as on the course design. © 2014 John Wiley & Sons Ltd.

  18. RERTR program progress in qualifying reduced-enrichment fuels

    International Nuclear Information System (INIS)

    Snelgrove, James L.

    1983-01-01

    In order to provide the technical means for reducing the enrichment of uranium used to fuel research and test reactors, the U.S. Reduced Enrichment Research and Test Reactor (RERTR) Program has been engaged in the development and testing of higher-uranium-density fuels than had been used previously. This fuel development effort included work to increase the density of fuels which were being used at the time the Program began and work on a fuel with the potential for much higher density. The ultimate goal of the fuel development and testing phase of the Program is to 'qualify' the fuel for use. A fuel is considered qualified when a sufficient data base for the fuel exists that it can be approved by regulating bodies for use in reactors. To convert a core to the use of reduced-enrichment fuel it is necessary to show that the core will behave properly during normal and off-normal operating conditions and to show that the fuel will behave properly to a reasonable margin beyond the conditions expected during normal operation. It is this latter area that this paper will address. The main characteristics to be considered in evaluating the performance of a fuel are its swelling, its blister-threshold temperature, and its metallurgical appearance. Data for the qualification of the reduced-enrichment fuels being developed by the RERTR Program are obtained from examination of miniature fuel plates (miniplates) which successfully pass the irradiation screening tests and from examinations of full-sized fuel elements. This paper will summarize the miniplate data reported in other papers presented during this meeting and will give the status of full-sized element irradiations. Finally, the current status of qualification of the various fuel types will be discussed and some projections of the future will be given

  19. Imaging and controlling intracellular reactions: Lysosome transport as a function of diameter and the intracellular synthesis of conducting polymers

    Science.gov (United States)

    Payne, Christine

    2014-03-01

    Eukaryotic cells are the ultimate complex environment with intracellular chemical reactions regulated by the local cellular environment. For example, reactants are sequestered into specific organelles to control local concentration and pH, motor proteins transport reactants within the cell, and intracellular vesicles undergo fusion to bring reactants together. Current research in the Payne Lab in the School of Chemistry and Biochemistry at Georgia Tech is aimed at understanding and utilizing this complex environment to control intracellular chemical reactions. This will be illustrated using two examples, intracellular transport as a function of organelle diameter and the intracellular synthesis of conducting polymers. Using single particle tracking fluorescence microscopy, we measured the intracellular transport of lysosomes, membrane-bound organelles, as a function of diameter as they underwent transport in living cells. Both ATP-dependent active transport and diffusion were examined. As expected, diffusion scales with the diameter of the lysosome. However, active transport is unaffected suggesting that motor proteins are insensitive to cytosolic drag. In a second example, we utilize intracellular complexity, specifically the distinct micro-environments of different organelles, to carry out chemical reactions. We show that catalase, found in the peroxisomes of cells, can be used to catalyze the polymerization of the conducting polymer PEDOT:PSS. More importantly, we have found that a range of iron-containing biomolecules are suitable catalysts with different iron-containing biomolecules leading to different polymer properties. These experiments illustrate the advantage of intracellular complexity for the synthesis of novel materials.

  20. Intracellular Localization of Arabidopsis Sulfurtransferases1

    Science.gov (United States)

    Bauer, Michael; Dietrich, Christof; Nowak, Katharina; Sierralta, Walter D.; Papenbrock, Jutta

    2004-01-01

    Sulfurtransferases (Str) comprise a group of enzymes widely distributed in archaea, eubacteria, and eukaryota which catalyze the transfer of a sulfur atom from suitable sulfur donors to nucleophilic sulfur acceptors. In all organisms analyzed to date, small gene families encoding Str proteins have been identified. The gene products were localized to different compartments of the cells. Our interest concerns the localization of Str proteins encoded in the nuclear genome of Arabidopsis. Computer-based prediction methods revealed localization in different compartments of the cell for six putative AtStrs. Several methods were used to determine the localization of the AtStr proteins experimentally. For AtStr1, a mitochondrial localization was demonstrated by immunodetection in the proteome of isolated mitochondria resolved by one- and two-dimensional gel electrophoresis and subsequent blotting. The respective mature AtStr1 protein was identified by mass spectrometry sequencing. The same result was obtained by transient expression of fusion constructs with the green fluorescent protein in Arabidopsis protoplasts, whereas AtStr2 was exclusively localized to the cytoplasm by this method. Three members of the single-domain AtStr were localized in the chloroplasts as demonstrated by transient expression of green fluorescent protein fusions in protoplasts and stomata, whereas the single-domain AtStr18 was shown to be cytoplasmic. The remarkable subcellular distribution of AtStr15 was additionally analyzed by transmission electron immunomicroscopy using a monospecific antibody against green fluorescent protein, indicating an attachment to the thylakoid membrane. The knowledge of the intracellular localization of the members of this multiprotein family will help elucidate their specific functions in the organism. PMID:15181206

  1. Intracellular signaling by diffusion: can waves of hydrogen peroxide transmit intracellular information in plant cells?

    DEFF Research Database (Denmark)

    Vestergaard, Christian L.; Flyvbjerg, Henrik; Møller, Ian Max

    2012-01-01

    of the physical and biochemical conditions in plant cells. As model system, we use a H(2)O(2) signal originating at the plasma membrane (PM) and spreading through the cytosol. We consider two maximally simple types of signals, isolated pulses and harmonic oscillations. First we consider the basic limits......Amplitude- and frequency-modulated waves of Ca(2+) ions transmit information inside cells. Reactive Oxygen Species (ROS), specifically hydrogen peroxide, have been proposed to have a similar role in plant cells. We consider the feasibility of such an intracellular communication system in view...

  2. New perspective in the assessment of total intracellular magnesium

    Directory of Open Access Journals (Sweden)

    Azzurra Sargenti

    2014-01-01

    Full Text Available Magnesium (Mg is essential for biological processes, but its cellular homeostasis has not been thoroughly elucidated, mainly because of the inadequacy of the available techniques to map intracellular Mg distribution. Recently, particular interest has been raised by a new family of fluorescent probes, diaza-18-crown-hydroxyquinoline (DCHQ, that shows remarkably high affinity and specificity for Mg, thus permitting the detection of the total intracellular Mg. The data obtained by fluori- metric and cytofluorimetric assays performed with DCHQ5 are in good agreement with atomic absorption spectroscopy, confirming that DCHQ5 probe allows both qualitative and quantitative determination of total intracellular Mg.

  3. Newly qualified teachers' visions of science learning and teaching

    Science.gov (United States)

    Roberts, Deborah L.

    2011-12-01

    This study investigated newly qualified teachers' visions of science learning and teaching. The study also documented their preparation in an elementary science methods course. The research questions were: What educational and professional experiences influenced the instructor's visions of science learning and teaching? What visions of science learning and teaching were promoted in the participants' science methods course? What visions of science learning and teaching did these newly qualified teachers bring with them as they graduated from their teacher preparation program? How did these visions compare with those advocated by reform documents? Data sources included participants' assignments, weekly reflections, and multi-media portfolio finals. Semi-structured interviews provided the emic voice of participants, after graduation but before they had begun to teach. These data were interpreted via a combination of qualitative methodologies. Vignettes described class activities. Assertions supported by excerpts from participants' writings emerged from repeated review of their assignments. A case study of a typical participant characterized weekly reflections and final multi-media portfolio. Four strands of science proficiency articulated in a national reform document provided a framework for interpreting activities, assignments, and interview responses. Prior experiences that influenced design of the methods course included an inquiry-based undergraduate physics course, participation in a reform-based teacher preparation program, undergraduate and graduate inquiry-based science teaching methods courses, participation in a teacher research group, continued connection to the university as a beginning teacher, teaching in diverse Title 1 schools, service as the county and state elementary science specialist, participation in the Carnegie Academy for the Scholarship of Teaching and Learning, service on a National Research Council committee, and experience teaching a

  4. Intracellular trafficking of new anticancer therapeutics: antibody–drug conjugates

    Science.gov (United States)

    Kalim, Muhammad; Chen, Jie; Wang, Shenghao; Lin, Caiyao; Ullah, Saif; Liang, Keying; Ding, Qian; Chen, Shuqing; Zhan, Jinbiao

    2017-01-01

    Antibody–drug conjugate (ADC) is a milestone in targeted cancer therapy that comprises of monoclonal antibodies chemically linked to cytotoxic drugs. Internalization of ADC takes place via clathrin-mediated endocytosis, caveolae-mediated endocytosis, and pinocytosis. Conjugation strategies, endocytosis and intracellular trafficking optimization, linkers, and drugs chemistry present a great challenge for researchers to eradicate tumor cells successfully. This inventiveness of endocytosis and intracellular trafficking has given considerable momentum recently to develop specific antibodies and ADCs to treat cancer cells. It is significantly advantageous to emphasize the endocytosis and intracellular trafficking pathways efficiently and to design potent engineered conjugates and biological entities to boost efficient therapies enormously for cancer treatment. Current studies illustrate endocytosis and intracellular trafficking of ADC, protein, and linker strategies in unloading and also concisely evaluate practically applicable ADCs. PMID:28814834

  5. Intracellular trafficking of new anticancer therapeutics: antibody-drug conjugates.

    Science.gov (United States)

    Kalim, Muhammad; Chen, Jie; Wang, Shenghao; Lin, Caiyao; Ullah, Saif; Liang, Keying; Ding, Qian; Chen, Shuqing; Zhan, Jinbiao

    2017-01-01

    Antibody-drug conjugate (ADC) is a milestone in targeted cancer therapy that comprises of monoclonal antibodies chemically linked to cytotoxic drugs. Internalization of ADC takes place via clathrin-mediated endocytosis, caveolae-mediated endocytosis, and pinocytosis. Conjugation strategies, endocytosis and intracellular trafficking optimization, linkers, and drugs chemistry present a great challenge for researchers to eradicate tumor cells successfully. This inventiveness of endocytosis and intracellular trafficking has given considerable momentum recently to develop specific antibodies and ADCs to treat cancer cells. It is significantly advantageous to emphasize the endocytosis and intracellular trafficking pathways efficiently and to design potent engineered conjugates and biological entities to boost efficient therapies enormously for cancer treatment. Current studies illustrate endocytosis and intracellular trafficking of ADC, protein, and linker strategies in unloading and also concisely evaluate practically applicable ADCs.

  6. EVIDENCE FOR THE MACROPHAGE INDUCING GENE IN MYCOBACTERIUM INTRACELLULARE

    Science.gov (United States)

    Background: The Mycobacterium avium Complex (MAC) includes the species M. avium (MA), M. intracellulare (MI), and possibly others. Organisms belonging to the MAC are phylogenetically closely related, opportunistic pathogens. The macrophage inducing gene (mig) is the only well-des...

  7. Data for automated, high-throughput microscopy analysis of intracellular bacterial colonies using spot detection

    DEFF Research Database (Denmark)

    Ernstsen, Christina Lundgaard; Login, Frédéric H.; Jensen, Helene Halkjær

    2017-01-01

    Quantification of intracellular bacterial colonies is useful in strategies directed against bacterial attachment, subsequent cellular invasion and intracellular proliferation. An automated, high-throughput microscopy-method was established to quantify the number and size of intracellular bacteria...

  8. Intracellular sodium hydrogen exchange inhibition and clinical myocardial protection.

    Science.gov (United States)

    Mentzer, Robert M; Lasley, Robert D; Jessel, Andreas; Karmazyn, Morris

    2003-02-01

    Although the mechanisms underlying ischemia/reperfusion injury remain elusive, evidence supports the etiologic role of intracellular calcium overload and oxidative stress induced by reactive oxygen species. Activation of the sodium hydrogen exchanger (NHE) is associated with intracellular calcium accumulation. Inhibition of the NHE-1 isoform may attenuate the consequences of this injury. Although there is strong preclinical and early clinical evidence that NHE inhibitors may be cardioprotective, definitive proof of this concept in humans awaits the results of ongoing clinical trials.

  9. 26 CFR 1.1092(c)-3 - Qualifying over-the-counter options.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 11 2010-04-01 2010-04-01 true Qualifying over-the-counter options. 1.1092(c)-3...-counter options. (a) In general. Under section 1092(c)(4)(B)(i), an equity option is not a qualified covered call option unless it is traded on a national securities exchange that is registered with the...

  10. Predictors of Self-Directed Learning for Low-Qualified Employees: A Multi-Level Analysis

    Science.gov (United States)

    Raemdonck, Isabel; van der Leeden, Rien; Valcke, Martin; Segers, Mien; Thijssen, Jo

    2012-01-01

    Purpose: This study aims to examine which variables at the level of the individual employee and at the company level are predictors of self-directed learning in low-qualified employees. Methodology: Results were obtained from a sample of 408 low-qualified employees from 35 different companies. The companies were selected from the energy sector,…

  11. 26 CFR 20.2056A-2 - Requirements for qualified domestic trust.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 14 2010-04-01 2010-04-01 false Requirements for qualified domestic trust. 20.2056A-2 Section 20.2056A-2 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) ESTATE AND GIFT TAXES ESTATE TAX; ESTATES OF DECEDENTS DYING AFTER AUGUST 16, 1954 Taxable Estate § 20.2056A-2 Requirements for qualified...

  12. 29 CFR 779.382 - May qualify as exempt 13(a)(2) establishments.

    Science.gov (United States)

    2010-07-01

    ... STANDARDS ACT AS APPLIED TO RETAILERS OF GOODS OR SERVICES Exemptions for Certain Retail or Service Establishments Hotels and Motels § 779.382 May qualify as exempt 13(a)(2) establishments. A hotel or motel establishment may qualify as an exempt retail or service establishment under section 13(a)(2) of the Act...

  13. 77 FR 76941 - Defense Federal Acquisition Regulation Supplement: New Qualifying Country-Poland (DFARS Case 2012...

    Science.gov (United States)

    2012-12-31

    ... discriminatory barriers to procurements of supplies and services produced by industrial enterprises of the other....003 is amended in paragraph (10), the definition of ``qualifying country'' by adding in alphabetical... place; 0 b. In paragraph (a), the definition of ``qualifying country'', by adding, in alphabetical order...

  14. The Learning Intentions of Low-Qualified Employees: A Multilevel Approach

    Science.gov (United States)

    Kyndt, Eva; Dochy, Filip; Onghena, Patrick; Baert, Herman

    2013-01-01

    In the continuously changing society and knowledge-intensive economy, the demand for the recurrent updating of competencies is coming to the fore for all employees, including low-qualified employees. Employees are considered low qualified when they do not have a starter qualification for higher education. Although many educational studies have…

  15. 7 CFR 785.8 - Reports by qualifying States receiving mediation grant funds.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 7 2010-01-01 2010-01-01 false Reports by qualifying States receiving mediation grant... SERVICE AGENCY, DEPARTMENT OF AGRICULTURE SPECIAL PROGRAMS CERTIFIED STATE MEDIATION PROGRAM § 785.8 Reports by qualifying States receiving mediation grant funds. (a) Annual report by certified State...

  16. 26 CFR 1.127-1 - Amounts received under a qualified educational assistance program.

    Science.gov (United States)

    2010-04-01

    ... employee under a qualified educational assistance program described in § 1.127-2, or (2) The value of... Income § 1.127-1 Amounts received under a qualified educational assistance program. (a) Exclusion from.... (c) Amounts received under a nonqualified program. Any amount received under an educational...

  17. 48 CFR 225.872-7 - Industrial security for qualifying countries.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 3 2010-10-01 2010-10-01 false Industrial security for... Agreements and Coordination 225.872-7 Industrial security for qualifying countries. The required procedures... qualifying country sources are in the DoD Industrial Security Regulation DoD 5220.22-R (implemented for the...

  18. Creating Highly Qualified Teachers: Maximizing University Resources to Provide Professional Development in Rural Areas

    Science.gov (United States)

    Mollenkopf, Dawn L.

    2009-01-01

    The "highly qualified teacher" requirement of No Child Left Behind has put pressure on rural school districts to recruit and retain highly qualified regular and special education teachers. If necessary, they may utilize uncertified, rural teachers with provisional certification; however, these teachers may find completing the necessary…

  19. 26 CFR 1.883-4T - Qualified shareholder stock ownership test (temporary).

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 9 2010-04-01 2010-04-01 false Qualified shareholder stock ownership test... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Foreign Corporations § 1.883-4T Qualified shareholder stock... the stock; (D) If the individual directly owns an interest in a corporation, partnership, trust...

  20. 41 CFR 102-80.135 - Who is a qualified fire protection engineer?

    Science.gov (United States)

    2010-07-01

    ... protection engineer? 102-80.135 Section 102-80.135 Public Contracts and Property Management Federal Property... qualified fire protection engineer? A qualified fire protection engineer is defined as an individual with a thorough knowledge and understanding of the principles of physics and chemistry governing fire growth...

  1. 75 FR 1831 - Seeks Qualified Candidates for the Advisory Committee on Reactor Safeguards

    Science.gov (United States)

    2010-01-13

    ... NUCLEAR REGULATORY COMMISSION Seeks Qualified Candidates for the Advisory Committee on Reactor... Regulatory Commission (NRC) seeks qualified candidates for the Advisory Committee on Reactor Safeguards (ACRS... amended. ACRS provides independent expert advice on matters related to the safety of existing and proposed...

  2. 14 CFR 1214.1104 - Evaluation and ranking of highly qualified candidates.

    Science.gov (United States)

    2010-01-01

    ... 14 Aeronautics and Space 5 2010-01-01 2010-01-01 false Evaluation and ranking of highly qualified candidates. 1214.1104 Section 1214.1104 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION... ranking of highly qualified candidates. (a) A selection board consisting of discipline experts, and such...

  3. 12 CFR 617.7125 - How should a qualified lender determine the effective interest rate?

    Science.gov (United States)

    2010-01-01

    ... effective interest rate? 617.7125 Section 617.7125 Banks and Banking FARM CREDIT ADMINISTRATION FARM CREDIT SYSTEM BORROWER RIGHTS Disclosure of Effective Interest Rates § 617.7125 How should a qualified lender determine the effective interest rate? (a) A qualified lender must calculate the effective interest rate on...

  4. 7 CFR 636.18 - Technical services provided by qualified personnel not affiliated with USDA.

    Science.gov (United States)

    2010-01-01

    ... HABITAT INCENTIVES PROGRAM § 636.18 Technical services provided by qualified personnel not affiliated with... affiliated with USDA. 636.18 Section 636.18 Agriculture Regulations of the Department of Agriculture... provided by qualified personnel not affiliated with USDA may include, but is not limited to: Conservation...

  5. 7 CFR 1466.11 - Technical services provided by qualified personnel not affiliated with USDA.

    Science.gov (United States)

    2010-01-01

    ... not affiliated with USDA. 1466.11 Section 1466.11 Agriculture Regulations of the Department of... OPERATIONS ENVIRONMENTAL QUALITY INCENTIVES PROGRAM Contracts and Payments § 1466.11 Technical services provided by qualified personnel not affiliated with USDA. (a) NRCS may use the services of qualified TSPs...

  6. 13 CFR 126.204 - May a qualified HUBZone SBC have affiliates?

    Science.gov (United States)

    2010-01-01

    ... affiliates? 126.204 Section 126.204 Business Credit and Assistance SMALL BUSINESS ADMINISTRATION HUBZONE PROGRAM Requirements to be a Qualified HUBZone SBC § 126.204 May a qualified HUBZone SBC have affiliates? A concern may have affiliates provided that the aggregate size of the concern and all of its...

  7. 26 CFR 1.401-7 - Forfeitures under a qualified pension plan.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 5 2010-04-01 2010-04-01 false Forfeitures under a qualified pension plan. 1...) INCOME TAX (CONTINUED) INCOME TAXES Pension, Profit-Sharing, Stock Bonus Plans, Etc. § 1.401-7 Forfeitures under a qualified pension plan. (a) General rules. In the case of a trust forming a part of a...

  8. 12 CFR 574.8 - Qualified stock issuances by undercapitalized savings associations or holding companies.

    Science.gov (United States)

    2010-01-01

    ... voting stock of the issuing undercapitalized savings association or savings and loan holding company. (8... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Qualified stock issuances by undercapitalized... SUPERVISION, DEPARTMENT OF THE TREASURY ACQUISITION OF CONTROL OF SAVINGS ASSOCIATIONS § 574.8 Qualified stock...

  9. 26 CFR 1.989(a)-1 - Definition of a qualified business unit.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 10 2010-04-01 2010-04-01 false Definition of a qualified business unit. 1.989... definition of the term “qualified business unit” (QBU) within the meaning of section 989. (2) Effective date... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Export Trade Corporations § 1.989(a)-1 Definition of a...

  10. Newly qualified teachers´ possibilities to get foothold in a lifelong career course

    DEFF Research Database (Denmark)

    Krøjgaard, Frede; Frederiksen, Lisbeth Angela Lunde

    Keyword: Induction program, newly qualified teachers, NQT, retention, professional development In Contrary to many other countries in Europe Denmark does not have any kind of national program regarding teacher induction program (TIP) or support in general to newly qualified teachers what so ever...

  11. 26 CFR 1.414(r)-1 - Requirements applicable to qualified separate lines of business.

    Science.gov (United States)

    2010-04-01

    ... lines of business. 1.414(r)-1 Section 1.414(r)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT... Plans, Etc. § 1.414(r)-1 Requirements applicable to qualified separate lines of business. (a) In general. Section 414(r) prescribes the conditions under which an employer is treated as operating qualified...

  12. 8 CFR 214.14 - Alien victims of certain qualifying criminal activity.

    Science.gov (United States)

    2010-01-01

    ... 8 Aliens and Nationality 1 2010-01-01 2010-01-01 false Alien victims of certain qualifying criminal activity. 214.14 Section 214.14 Aliens and Nationality DEPARTMENT OF HOMELAND SECURITY IMMIGRATION REGULATIONS NONIMMIGRANT CLASSES § 214.14 Alien victims of certain qualifying criminal activity. (a...

  13. Qualified-agent virtue ethics | van Zyl | South African Journal of ...

    African Journals Online (AJOL)

    Qualified-agent virtue ethics provides an account of right action in terms of the virtuous agent. It has become one of the most popular, but also most frequently criticized versions of virtue ethics. Many of the objections rest on the mistaken assumption that proponents of qualified-agent virtue ethics share the same view when it ...

  14. 26 CFR 1.181-3T - Qualified film or television production (temporary).

    Science.gov (United States)

    2010-04-01

    ... season of programming. (3) Exception for certain sexually explicit productions. A production does not... 26 Internal Revenue 3 2010-04-01 2010-04-01 false Qualified film or television production... (continued) § 1.181-3T Qualified film or television production (temporary). (a) In general. The term...

  15. 26 CFR 1.1293-1 - Current taxation of income from qualified electing funds.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 11 2010-04-01 2010-04-01 true Current taxation of income from qualified electing funds. 1.1293-1 Section 1.1293-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE... Losses § 1.1293-1 Current taxation of income from qualified electing funds. (a) In general. [Reserved] (1...

  16. 78 FR 56872 - City of Barre, Vermont; Notice of Preliminary Determination of a Qualifying Conduit Hydropower...

    Science.gov (United States)

    2013-09-16

    ..., Vermont; Notice of Preliminary Determination of a Qualifying Conduit Hydropower Facility and Soliciting... construct a qualifying conduit hydropower facility, pursuant to section 30 of the Federal Power Act, as amended by section 4 of the Hydropower Regulatory Efficiency Act of 2013 (HREA). The Nelson Street 17 kW...

  17. 78 FR 66355 - Pleasant Grove City, UT; Notice of Preliminary Determination of a Qualifying Conduit Hydropower...

    Science.gov (United States)

    2013-11-05

    ... City, UT; Notice of Preliminary Determination of a Qualifying Conduit Hydropower Facility and... Grove City, Utah (Pleasant Grove) filed a notice of intent to construct a qualifying conduit hydropower facility, pursuant to section 30 of the Federal Power Act, as amended by section 4 of the Hydropower...

  18. 78 FR 61987 - Corbett Water District; Notice of Preliminary Determination of a Qualifying Conduit Hydropower...

    Science.gov (United States)

    2013-10-09

    ... District; Notice of Preliminary Determination of a Qualifying Conduit Hydropower Facility and Soliciting... construct a qualifying conduit hydropower facility, pursuant to section 30 of the Federal Power Act, as amended by section 4 of the Hydropower Regulatory Efficiency Act of 2013 (HREA). The Corbett Hydroelectric...

  19. 78 FR 53752 - City of Sandpoint, Idaho; Notice of Preliminary Determination of a Qualifying Conduit Hydropower...

    Science.gov (United States)

    2013-08-30

    ... Sandpoint, Idaho; Notice of Preliminary Determination of a Qualifying Conduit Hydropower Facility and... intent to construct a qualifying conduit hydropower facility, pursuant to section 30 of the Federal Power Act, as amended by section 4 of the Hydropower Regulatory Efficiency Act of 2013 (HREA). The Little...

  20. 78 FR 61985 - City of Astoria, Oregon; Notice of Preliminary Determination of a Qualifying Conduit Hydropower...

    Science.gov (United States)

    2013-10-09

    ..., Oregon; Notice of Preliminary Determination of a Qualifying Conduit Hydropower Facility and Soliciting... of intent to construct a qualifying conduit hydropower facility, pursuant to section 30 of the Federal Power Act, as amended by section 4 of the Hydropower Regulatory Efficiency Act of 2013 (HREA). The...

  1. 78 FR 63176 - Notice of Preliminary Determination of a Qualifying Conduit Hydropower Facility and Soliciting...

    Science.gov (United States)

    2013-10-23

    ... Preliminary Determination of a Qualifying Conduit Hydropower Facility and Soliciting Comments and Motions To... of intent to construct a qualifying conduit hydropower facility, pursuant to section 30 of the Federal Power Act, as amended by section 4 of the Hydropower Regulatory Efficiency Act of 2013 (HREA). The...

  2. 78 FR 62350 - FFP Qualified Hydro 14, LLC; Western Minnesota Municipal Power Agency; Notice Announcing...

    Science.gov (United States)

    2013-10-18

    .... 14491-000] FFP Qualified Hydro 14, LLC; Western Minnesota Municipal Power Agency; Notice Announcing... applications were filed by FFP Qualified Hydro 14, LLC for Project No. 13579-002 and Western Minnesota... regular business day. See 18 CFR 385.2001(a)(2) (2013). On October 21, 2013, at 10:00 a.m. (Eastern Time...

  3. Attempting to Predict Success in the Qualifying Round of the International Chemistry Olympiad

    Science.gov (United States)

    Urhahne, Detlef; Ho, Lok Hang; Parchmann, Ilka; Nick, Sabine

    2012-01-01

    The aim of this study was trying to predict success in the qualifying round for the International Chemistry Olympiad (IChO) on the basis of the expectancy-value model of achievement motivation by Eccles et al. The investigation with 52 participants, including 14 females, was conducted during the third of four qualifying rounds of the IChO in…

  4. 12 CFR 617.7130 - What initial disclosures must a qualified lender make to a borrower?

    Science.gov (United States)

    2010-01-01

    ... lender make to a borrower? 617.7130 Section 617.7130 Banks and Banking FARM CREDIT ADMINISTRATION FARM... must a qualified lender make to a borrower? (a) Required disclosures—in general. A qualified lender... certificates that borrowers are required to purchase are at risk and may only be retired at the discretion of...

  5. 26 CFR 1.401(a)-11 - Qualified joint and survivor annuities.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 5 2010-04-01 2010-04-01 false Qualified joint and survivor annuities. 1.401(a...)-11 Qualified joint and survivor annuities. (a) General rule—(1) Required provisions. A trust, to...), which is a part of a plan providing for the payment of benefits in any form of a life annuity (as...

  6. 26 CFR 11.401(a)-11 - Qualified joint and survivor annuities.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 14 2010-04-01 2010-04-01 false Qualified joint and survivor annuities. 11.401... SECURITY ACT OF 1974 § 11.401(a)-11 Qualified joint and survivor annuities. (a) In general—(1) General rule... annuity (i.e., an annuity requiring survival of the participant or his spouse as a condition for payment...

  7. 76 FR 2029 - Small Business Investment Companies-Energy Saving Qualified Investments

    Science.gov (United States)

    2011-01-12

    ... 3245-AF86 Small Business Investment Companies--Energy Saving Qualified Investments AGENCY: U.S. Small... Administration (SBA) is setting forth the new defined terms, ``Energy Saving Qualified Investment'' and ``Energy Saving Activities'', for the Small Business Investment Company (SBIC) Program. The new definitions are...

  8. Qualifying concrete for a low-level waste repository

    International Nuclear Information System (INIS)

    Philipose, K.E.

    1990-06-01

    A waste repository for the belowground disposal of low-level radioactive waste, labelled IRUS (Intrusion Resistant Underground Structure), is planned at Chalk River Nuclear Laboratories. It relies greatly on the durability of concrete for a minimum of 500 years of service life. A research program based on laboratory testing to design a durable concrete and predict its useful engineered service life is in progress. Durability of concrete depends on its resistance of deterioration from both internal and external causes. Since the rate of degradation depends to a major extent on the rate of ingress of aggressive ions into concrete, laboratory testing is in progress to establish the diffusion rates of ions, especially chlorides, sulphate and carbonate ions. A total of 1000 concrete specimens and 500 paste specimens are being exposed at 22 and 45 degrees C to twenty-five different combinations of corrosive agents, including CO 2 . Procedures to measure the ionic profile and to determine the factors controlling diffusion of ions in the various concretes have been developed. The paper presents the initial results from the research program and the longevity predictions to qualify concretes for the IRUS waste repository, based on twelve months of diffusion testing on laboratory specimens

  9. Life cycle evaluation of spaceflight qualified nickel-hydrogen batteries

    Energy Technology Data Exchange (ETDEWEB)

    Coates, D.K.; Brill, J.N. [Eagle-Picher Industries, Inc., Joplin, MO (United States). Advanced Systems Operation

    1995-12-31

    Life cycle test results are summarized from more than 300 spaceflight qualified nickel-hydrogen (NiH{sub 2}) battery cells currently on life test. Cells ranging in size from 4 ampere-hours (Ah) to 120 Ah are being tested under a variety of conditions to support current NiH{sub 2} battery applications. Results to date include 55,600 accelerated LEO cycles at 30% DOD; 102,840 accelerated LEO cycles at 15% DOD; 44,900 cycles under a real-time LEO profile; 44,100 cycles in real-time LEO; 30 accelerated GEO eclipse seasons and 7 real-time GEO eclipse seasons, both at 75% DOD maximum. Alternative separator materials have completed more than 40,000 charge/discharge cycles in accelerated LEO testing and advanced design electrocatalytic hydrogen electrodes have completed more than 16,000 cycles in real-time LEO testing. Common pressure vessel cell designs have completed 18,000 cycles in real-time LEO testing at 45% DOD.

  10. Gender Inequalities in Highly Qualified Professions: A Social Psychological Analysis

    Directory of Open Access Journals (Sweden)

    Maria Helena Santos

    2016-06-01

    Full Text Available Research in social and political psychology contributes towards understanding the persistence of job market gender segregation prevailing in recent decades, the consequences for those involved and their reactions when having to cope with gender inequality. Within the framework of the literature on shared ideologies that justify and legitimize discrimination against women, this article focuses on Portugal and analyses the particular case of women in two highly qualified professions traditionally carried out by men – politics and medicine. Drawing on the results of quantitative and qualitative studies, our analytical approach demonstrates how while a majority of participants show awareness of the existence of gender inequality in these markedly masculine professions, meritocratic individualism and personal attributions to discrimination are the recurring explanations rather than any gender-based account. These results allow us to highlight the relevance of gender-based analysis as an ideology and furthermore to argue that ignoring this perspective not only diminishes individual responsibility for social change but also perpetuates gender asymmetries.

  11. Uptake and intracellular activity of AM-1155 in phagocytic cells.

    Science.gov (United States)

    Yamamoto, T; Kusajima, H; Hosaka, M; Fukuda, H; Oomori, Y; Shinoda, H

    1996-01-01

    The uptake and intracellular activity of AM-1155 in murine J774.1 macrophages and human polymorphonuclear leukocytes were investigated. AM-1155 penetrated phagocytic cells rapidly and reversibly, although the penetration process was not affected by metabolic inhibitors such as sodium fluoride, cyanide m-chlorophenylhydrazone, or ouabain or by nucleoside transport system inhibitors such as adenosine. The intracellular concentration-to-extracellular concentration ratio of AM-1155 in both cell types of phagocytes ranged from 5 to 7. These ratios were almost equal to those for sparfloxacin. The intracellular activity of AM-1155 in J774.1 macrophages, examined with Staphylococcus aureus 209P as a test bacterium, was dependent on the extracellular concentration. AM-1155 at a concentration of 1 microgram/ml reduced the number of viable cells of S. aureus ingested by more than 90%. The intracellular activity of AM-1155 was more potent than those of sparfloxacin, ofloxacin, ciprofloxacin, flomoxef, and erythromycin. These results suggest that the potent intracellular activity of AM-1155 might mainly be due to the high intracellular concentration and its potent in vitro activity. PMID:9124835

  12. Epithelial Cell Gene Expression Induced by Intracellular Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Xianglu Li

    2009-01-01

    Full Text Available HEp-2 cell monolayers were cocultured with intracellular Staphylococcus aureus, and changes in gene expression were profiled using DNA microarrays. Intracellular S. aureus affected genes involved in cellular stress responses, signal transduction, inflammation, apoptosis, fibrosis, and cholesterol biosynthesis. Transcription of stress response and signal transduction-related genes including atf3, sgk, map2k1, map2k3, arhb, and arhe was increased. In addition, elevated transcription of proinflammatory genes was observed for tnfa, il1b, il6, il8, cxcl1, ccl20, cox2, and pai1. Genes involved in proapoptosis and fibrosis were also affected at transcriptional level by intracellular S. aureus. Notably, intracellular S. aureus induced strong transcriptional down-regulation of several cholesterol biosynthesis genes. These results suggest that epithelial cells respond to intracellular S. aureus by inducing genes affecting immunity and in repairing damage caused by the organism, and are consistent with the possibility that the organism exploits an intracellular environment to subvert host immunity and promote colonization.

  13. Federally qualified health center dental clinics: financial information.

    Science.gov (United States)

    Bailit, Howard L; Devitto, Judy; Myne-Joslin, Ronnie; Beazoglou, Tryfon; McGowan, Taegan

    2013-01-01

    Federally Qualified Health Center (FQHC) dental clinics are a major component of the dental safety net system, providing care to 3.75 million patients annually. This study describes the financial and clinical operations of a sample of FQHCs. In cooperation with the National Network for Oral Health Access, FQHC dental clinics that could provide 12 months of electronic dental record information were asked to participate in the study. Based on data from 28 dental clinics (14 FQHCs), 50 percent of patients were under 21 years of age. The primary payers were Medicaid (72.4 percent) and sliding-scale/self-pay patients (17.5 percent). Sites averaged 3.1 operatories, 0.66 dental hygienists, and 1.9 other staff per dentist. Annually, each FTE dentist and hygienist provided 2,801 and 2,073 patient visits, respectively. Eighty percent of services were diagnostic, preventive, and restorative. Patient care accounted for 82 percent of revenues, and personnel (64.2 percent) and central administration (13.4 percent) accounted for most expenses. Based on a small convenience sample of FQHC dental clinics, this study presents descriptive data on their clinical and financial operations. Compared with data from the UDS (Uniform Data System) report, study FQHCs were larger in terms of space, staff, and patients served. However, there was substantial variation among clinics for almost all measures. As the number and size of FQHC dental clinics increase, the Health Resources and Services Administration needs to provide them access to comparative data that they can use to benchmark their operations. © 2013 American Association of Public Health Dentistry.

  14. An experience of qualified preventive screening: shiraz smart screening software.

    Science.gov (United States)

    Islami Parkoohi, Parisa; Zare, Hashem; Abdollahifard, Gholamreza

    2015-01-01

    Computerized preventive screening software is a cost effective intervention tool to address non-communicable chronic diseases. Shiraz Smart Screening Software (SSSS) was developed as an innovative tool for qualified screening. It allows simultaneous smart screening of several high-burden chronic diseases and supports reminder notification functionality. The extent in which SSSS affects screening quality is also described. Following software development, preventive screening and annual health examinations of 261 school staff (Medical School of Shiraz, Iran) was carried out in a software-assisted manner. To evaluate the quality of the software-assisted screening, we used quasi-experimental study design and determined coverage, irregular attendance and inappropriateness proportions in relation with the manual and software-assisted screening as well as the corresponding number of requested tests. In manual screening method, 27% of employees were covered (with 94% irregular attendance) while by software-assisted screening, the coverage proportion was 79% (attendance status will clear after the specified time). The frequency of inappropriate screening test requests, before the software implementation, was 41.37% for fasting plasma glucose, 41.37% for lipid profile, 0.84% for occult blood, 0.19% for flexible sigmoidoscopy/colonoscopy, 35.29% for Pap smear, 19.20% for mammography and 11.2% for prostate specific antigen. All of the above were corrected by the software application. In total, 366 manual screening and 334 software-assisted screening tests were requested. SSSS is an innovative tool to improve the quality of preventive screening plans in terms of increased screening coverage, reduction in inappropriateness and the total number of requested tests.

  15. Intracellular disposition of chitosan nanoparticles in macrophages: intracellular uptake, exocytosis, and intercellular transport

    Directory of Open Access Journals (Sweden)

    Jiang LQ

    2017-08-01

    Full Text Available Li Qun Jiang,1 Ting Yu Wang,1 Thomas J Webster,2 Hua-Jian Duan,1 Jing Ying Qiu,1 Zi Ming Zhao,1 Xiao Xing Yin,1,* Chun Li Zheng3,* 1Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, School of Pharmacy, Xuzhou Medical University, Xuzhou, People’s Republic of China; 2Department of Chemical Engineering, Northeastern University, Boston, MA, USA; 3School of Pharmacy, China Pharmaceutical University, Nanjing, People’s Republic of China *These authors contributed equally to this work Abstract: Biodegradable nanomaterials have been widely used in numerous medical fields. To further improve such efforts, this study focused on the intracellular disposition of chitosan nanoparticles (CsNPs in macrophages, a primary cell of the mononuclear phagocyte system (MPS. Such interactions with the MPS determine the nanoparticle retention time in the body and consequently play a significant role in their own clinical safety. In this study, various dye-labeled CsNPs (about 250 nm were prepared, and a murine macrophage cell line (RAW 264.7 was selected as a model macrophage. The results showed two mechanisms of macrophage incorporation of CsNPs, ie, a clathrin-mediated endocytosis pathway (the primary and phagocytosis. Following internalization, the particles partly dissociated in the cells, indicating cellular digestion of the nanoparticles. It was proved that, after intracellular uptake, a large proportion of CsNPs were exocytosed within 24 h; this excretion induced a decrease in fluorescence intensity in cells by 69%, with the remaining particles possessing difficulty being cleared. Exocytosis could be inhibited by both wortmannin and vacuolin-1, indicating that CsNP uptake was mediated by lysosomal and multivesicular body pathways, and after exocytosis, the reuptake of CsNPs by neighboring cells was verified by further experiments. This study, thus, elucidated the fate of CsNPs in macrophages as well as identified cellular disposition

  16. 'Practising under your own Pin'- a description of the transition experiences of newly qualified midwives.

    Science.gov (United States)

    Avis, Mark; Mallik, Maggie; Fraser, Diane M

    2013-11-01

    Transition experiences of newly qualified midwives were examined in depth during the third phase of a UK evaluation study of midwifery education. The fitness to practise and the retention of newly qualified nursing and midwifery graduates are pressing concerns for health care managers. The advantages of preceptorship are reported in the literature but the content and timing of schemes remain unclear. A semi-structured diary was kept for up to 6 months by 35 newly qualified midwives in 18 work sites covering all countries in the UK. The preceptor and supervisor of midwives for each newly qualified midwife completed short questionnaires about their preceptee's performance, and a further sub-sample of newly qualified midwives and preceptors participated in a semi-structured interview. Data were analysed to elicit aspects of newly qualified midwives transition experiences. Findings confirm that structured preceptorship schemes are not widely available. Newly qualified midwives primarily obtained transition support from members of the midwifery team. Although perceived as competent, there is no demarcation point in becoming confident to practise as a registered practitioner. Implications for managers include the importance of a supportive culture within clinical teams for successful transition and the introduction of structured preceptorship schemes facilitated by appropriate rotation patterns. © 2012 John Wiley & Sons Ltd.

  17. Relevance of intracellular polarity to accuracy of eukaryotic chemotaxis

    International Nuclear Information System (INIS)

    Hiraiwa, Tetsuya; Nishikawa, Masatoshi; Shibata, Tatsuo; Nagamatsu, Akihiro; Akuzawa, Naohiro

    2014-01-01

    Eukaryotic chemotaxis is usually mediated by intracellular signals that tend to localize at the front or back of the cell. Such intracellular polarities frequently require no extracellular guidance cues, indicating that spontaneous polarization occurs in the signal network. Spontaneous polarization activity is considered relevant to the persistent motions in random cell migrations and chemotaxis. In this study, we propose a theoretical model that connects spontaneous intracellular polarity and motile ability in a chemoattractant solution. We demonstrate that the intracellular polarity can enhance the accuracy of chemotaxis. Chemotactic accuracy should also depend on chemoattractant concentration through the concentration-dependent correlation time in the polarity direction. Both the polarity correlation time and the chemotactic accuracy depend on the degree of responsiveness to the chemical gradient. We show that optimally accurate chemotaxis occurs at an intermediate responsiveness of intracellular polarity. Experimentally, we find that the persistence time of randomly migrating Dictyostelium cells depends on the chemoattractant concentration, as predicted by our theory. At the optimum responsiveness, this ameboid cell can enhance its chemotactic accuracy tenfold. (paper)

  18. Intracellular calcium levels can regulate Importin-dependent nuclear import

    International Nuclear Information System (INIS)

    Kaur, Gurpreet; Ly-Huynh, Jennifer D.; Jans, David A.

    2014-01-01

    Highlights: • High intracellular calcium inhibits Impα/β1- or Impβ1-dependent nuclear protein import. • The effect of Ca 2+ on nuclear import does not relate to changes in the nuclear pore. • High intracellular calcium can result in mislocalisation of Impβ1, Ran and RCC1. - Abstract: We previously showed that increased intracellular calcium can modulate Importin (Imp)β1-dependent nuclear import of SRY-related chromatin remodeling proteins. Here we extend this work to show for the first time that high intracellular calcium inhibits Impα/β1- or Impβ1-dependent nuclear protein import generally. The basis of this relates to the mislocalisation of the transport factors Impβ1 and Ran, which show significantly higher nuclear localization in contrast to various other factors, and RCC1, which shows altered subnuclear localisation. The results here establish for the first time that intracellular calcium modulates conventional nuclear import through direct effects on the nuclear transport machinery

  19. Intracellular transport of fat-soluble vitamins A and E.

    Science.gov (United States)

    Kono, Nozomu; Arai, Hiroyuki

    2015-01-01

    Vitamins are compounds that are essential for the normal growth, reproduction and functioning of the human body. Of the 13 known vitamins, vitamins A, D, E and K are lipophilic compounds and are therefore called fat-soluble vitamins. Because of their lipophilicity, fat-soluble vitamins are solubilized and transported by intracellular carrier proteins to exert their actions and to be metabolized properly. Vitamin A and its derivatives, collectively called retinoids, are solubilized by intracellular retinoid-binding proteins such as cellular retinol-binding protein (CRBP), cellular retinoic acid-binding protein (CRABP) and cellular retinal-binding protein (CRALBP). These proteins act as chaperones that regulate the metabolism, signaling and transport of retinoids. CRALBP-mediated intracellular retinoid transport is essential for vision in human. α-Tocopherol, the main form of vitamin E found in the body, is transported by α-tocopherol transfer protein (α-TTP) in hepatic cells. Defects of α-TTP cause vitamin E deficiency and neurological disorders in humans. Recently, it has been shown that the interaction of α-TTP with phosphoinositides plays a critical role in the intracellular transport of α-tocopherol and is associated with familial vitamin E deficiency. In this review, we summarize the mechanisms and biological significance of the intracellular transport of vitamins A and E. © 2014 The Authors. Traffic published by John Wiley & Sons Ltd.

  20. Exploring anti-bacterial compounds against intracellular Legionella.

    Directory of Open Access Journals (Sweden)

    Christopher F Harrison

    Full Text Available Legionella pneumophila is a ubiquitous fresh-water bacterium which reproduces within its erstwhile predators, environmental amoeba, by subverting the normal pathway of phagocytosis and degradation. The molecular mechanisms which confer resistance to amoeba are apparently conserved and also allow replication within macrophages. Thus, L. pneumophila can act as an 'accidental' human pathogen and cause a severe pneumonia known as Legionnaires' disease. The intracellular localisation of L. pneumophila protects it from some antibiotics, and this fact must be taken into account to develop new anti-bacterial compounds. In addition, the intracellular lifestyle of L. pneumophila may render the bacteria susceptible to compounds diminishing bacterial virulence and decreasing intracellular survival and replication of this pathogen. The development of a single infection cycle intracellular replication assay using GFP-producing L. pneumophila and Acanthamoebacastellanii amoeba is reported here. This fluorescence-based assay allows for continuous monitoring of intracellular replication rates, revealing the effect of bacterial gene deletions or drug treatment. To examine how perturbations of the host cell affect L. pneumophila replication, several known host-targeting compounds were tested, including modulators of cytoskeletal dynamics, vesicle scission and Ras GTPase localisation. Our results reveal a hitherto unrealized potential antibiotic property of the β-lactone-based Ras depalmitoylation inhibitor palmostatin M, but not the closely related inhibitor palmostatin B. Further characterisation indicated that this compound caused specific growth inhibition of Legionella and Mycobacterium species, suggesting that it may act on a common bacterial target.

  1. Surveillance for Intracellular Antibody by Cytosolic Fc Receptor TRIM21

    Directory of Open Access Journals (Sweden)

    William A. McEwan

    2016-11-01

    Full Text Available TRIM21 has emerged as an atypical Fc receptor that is broadly conserved and widely expressed in the cytoplasm of mammalian cells. Viruses that traffic surface-bound antibodies into the cell during infection recruit TRIM21 via a high affinity interaction between Fc and TRIM21 PRYSPRY domain. Following binding of intracellular antibody, TRIM21 acts as both antiviral effector and sensor for innate immune signalling. These activities serve to reduce viral replication by orders of magnitude in vitro and contribute to host survival during in vivo infection. Neutralization occurs rapidly after detection and requires the activity of the ubiquitin-proteasome system. The microbial targets of this arm of intracellular immunity are still being identified: TRIM21 activity has been reported following infection by several non-enveloped viruses and intracellular bacteria. These findings extend the sphere of influence of antibodies to the intracellular domain and have broad implications for immunity. TRIM21 has been implicated in the chronic auto-immune condition systemic lupus erythematosus and is itself an auto-antigen in Sjögren’s syndrome. This review summarises our current understanding of TRIM21’s role as a cytosolic Fc receptor and briefly discusses pathological circumstances where intracellular antibodies have been described, or are hypothesized to occur, and may benefit from further investigations of the role of TRIM21.

  2. Advances in genetic manipulation of obligate intracellular bacterial pathogens

    Directory of Open Access Journals (Sweden)

    Paul eBeare

    2011-05-01

    Full Text Available Infections by obligate intracellular bacterial pathogens result in significant morbidity and mortality worldwide. These bacteria include Chlamydia spp., which causes millions of cases of sexually transmitted disease and blinding trachoma annually, and members of the α-proteobacterial genera Anaplasma, Ehrlichia, Orientia and Rickettsia, agents of serious human illnesses including epidemic typhus. Coxiella burnetii, the agent of human Q fever, has also been considered a prototypical obligate intracellular bacterium, but recent host cell-free (axenic growth has rescued it from obligatism. The historic genetic intractability of obligate intracellular bacteria has severely limited molecular dissection of their unique lifestyles and virulence factors involved in pathogenesis. Host cell restricted growth is a significant barrier to genetic transformation that can make simple procedures for free-living bacteria, such as cloning, exceedingly difficult. Low transformation efficiency requiring long term culture in host cells to expand small transformant populations is another obstacle. Despite numerous technical limitations, the last decade has witnessed significant gains in genetic manipulation of obligate intracellular bacteria including allelic exchange. Continued development of genetic tools should soon enable routine mutation and complementation strategies for virulence factor discovery and stimulate renewed interest in these refractory pathogens. In this review, we discuss the technical challenges associated with genetic transformation of obligate intracellular bacteria and highlight advances made with individual genera.

  3. Exploring Anti-Bacterial Compounds against Intracellular Legionella

    Science.gov (United States)

    Harrison, Christopher F.; Kicka, Sébastien; Trofimov, Valentin; Berschl, Kathrin; Ouertatani-Sakouhi, Hajer; Ackermann, Nikolaus; Hedberg, Christian; Cosson, Pierre; Soldati, Thierry; Hilbi, Hubert

    2013-01-01

    Legionella pneumophila is a ubiquitous fresh-water bacterium which reproduces within its erstwhile predators, environmental amoeba, by subverting the normal pathway of phagocytosis and degradation. The molecular mechanisms which confer resistance to amoeba are apparently conserved and also allow replication within macrophages. Thus, L. pneumophila can act as an ‘accidental’ human pathogen and cause a severe pneumonia known as Legionnaires’ disease. The intracellular localisation of L. pneumophila protects it from some antibiotics, and this fact must be taken into account to develop new anti-bacterial compounds. In addition, the intracellular lifestyle of L. pneumophila may render the bacteria susceptible to compounds diminishing bacterial virulence and decreasing intracellular survival and replication of this pathogen. The development of a single infection cycle intracellular replication assay using GFP-producing L. pneumophila and Acanthamoeba castellanii amoeba is reported here. This fluorescence-based assay allows for continuous monitoring of intracellular replication rates, revealing the effect of bacterial gene deletions or drug treatment. To examine how perturbations of the host cell affect L. pneumophila replication, several known host-targeting compounds were tested, including modulators of cytoskeletal dynamics, vesicle scission and Ras GTPase localisation. Our results reveal a hitherto unrealized potential antibiotic property of the β-lactone-based Ras depalmitoylation inhibitor palmostatin M, but not the closely related inhibitor palmostatin B. Further characterisation indicated that this compound caused specific growth inhibition of Legionella and Mycobacterium species, suggesting that it may act on a common bacterial target. PMID:24058631

  4. Intracellular calcium levels can regulate Importin-dependent nuclear import

    Energy Technology Data Exchange (ETDEWEB)

    Kaur, Gurpreet; Ly-Huynh, Jennifer D.; Jans, David A., E-mail: David.Jans@monash.edu

    2014-07-18

    Highlights: • High intracellular calcium inhibits Impα/β1- or Impβ1-dependent nuclear protein import. • The effect of Ca{sup 2+} on nuclear import does not relate to changes in the nuclear pore. • High intracellular calcium can result in mislocalisation of Impβ1, Ran and RCC1. - Abstract: We previously showed that increased intracellular calcium can modulate Importin (Imp)β1-dependent nuclear import of SRY-related chromatin remodeling proteins. Here we extend this work to show for the first time that high intracellular calcium inhibits Impα/β1- or Impβ1-dependent nuclear protein import generally. The basis of this relates to the mislocalisation of the transport factors Impβ1 and Ran, which show significantly higher nuclear localization in contrast to various other factors, and RCC1, which shows altered subnuclear localisation. The results here establish for the first time that intracellular calcium modulates conventional nuclear import through direct effects on the nuclear transport machinery.

  5. Intracellular renin disrupts chemical communication between heart cells. Pathophysiological implications

    Directory of Open Access Journals (Sweden)

    Walmor eDe Mello

    2015-01-01

    Full Text Available The influence of intracellular renin on the process of chemical communication between cardiac cells was investigated in cell pairs isolated from the left ventricle of adult Wistar Kyoto rats. The enzyme together with Lucifer yellow CH was dialyzed into one cell of the pair using the whole cell clamp technique. The diffusion of the dye in the dialyzed and in non-dialyzed cell was followed by measuring the intensity of fluorescence in both cells as a function of time. The results indicated that; 1 under normal conditions, Lucifer Yellow flows from cell-to-cell through gap junctions; 2 the intracellular dialysis of renin (100nM disrupts chemical communication-an effect enhanced by simultaneous administration of angiotensinogen (100nM; 3 enalaprilat (10-9M administered to the cytosol together with renin reduced drastically the uncoupling action of the enzyme; 4 aliskiren (10-8M inhibited the effect of renin on chemical communication;5 the possible role of intracellular renin independently of angiotensin II (Ang II was evaluated including the increase of the inward calcium current elicited by the enzyme and the possible role of oxidative stress on the disruption of cell communication; 6 the possible harmful versus the beneficial effect of intracellular renin during myocardial infarction was discussed;7 the present results indicate that intracellular renin due to internalization or in situ synthesis, causes a severe impairment of chemical communication in the heart resulting in derangement of metabolic cooperation with serious consequences for heart function.

  6. Self-organization of intracellular gradients during mitosis

    Directory of Open Access Journals (Sweden)

    Fuller Brian G

    2010-01-01

    Full Text Available Abstract Gradients are used in a number of biological systems to transmit spatial information over a range of distances. The best studied are morphogen gradients where information is transmitted over many cell lengths. Smaller mitotic gradients reflect the need to organize several distinct events along the length of the mitotic spindle. The intracellular gradients that characterize mitosis are emerging as important regulatory paradigms. Intracellular gradients utilize intrinsic auto-regulatory feedback loops and diffusion to establish stable regions of activity within the mitotic cytosol. We review three recently described intracellular mitotic gradients. The Ran GTP gradient with its elaborate cascade of nuclear transport receptors and cargoes is the best characterized, yet the dynamics underlying the robust gradient of Ran-GTP have received little attention. Gradients of phosphorylation have been observed on Aurora B kinase substrates both before and after anaphase onset. In both instances the phosphorylation gradient appears to result from a soluble gradient of Aurora B kinase activity. Regulatory properties that support gradient formation are highlighted. Intracellular activity gradients that regulate localized mitotic events bare several hallmarks of self-organizing biologic systems that designate spatial information during pattern formation. Intracellular pattern formation represents a new paradigm in mitotic regulation.

  7. Quantifying intracellular hydrogen peroxide perturbations in terms of concentration

    Directory of Open Access Journals (Sweden)

    Beijing K. Huang

    2014-01-01

    Full Text Available Molecular level, mechanistic understanding of the roles of reactive oxygen species (ROS in a variety of pathological conditions is hindered by the difficulties associated with determining the concentration of various ROS species. Here, we present an approach that converts fold-change in the signal from an intracellular sensor of hydrogen peroxide into changes in absolute concentration. The method uses extracellular additions of peroxide and an improved biochemical measurement of the gradient between extracellular and intracellular peroxide concentrations to calibrate the intracellular sensor. By measuring peroxiredoxin activity, we found that this gradient is 650-fold rather than the 7–10-fold that is widely cited. The resulting calibration is important for understanding the mass-action kinetics of complex networks of redox reactions, and it enables meaningful characterization and comparison of outputs from endogenous peroxide generating tools and therapeutics across studies.

  8. Simple Recovery of Intracellular Gold Nanoparticles from Peanut Seedling Roots.

    Science.gov (United States)

    Raju, D; Mehta, Urmil J; Ahmad, Absar

    2015-02-01

    Fabrication of inorganic nanomaterials via a biological route witnesses the formation either extracellularly, intracellulary or both. Whereas extracellular formation of these nanomaterials is cherished owing to their easy and economical extraction and purification processes; the intracellular formation of nanomaterials, due to the lack of a proper recovery protocol has always been dreaded, as the extraction processes used so far were tedious, costly, time consuming and often resulting in very low recovery. The aim of the present study was to overcome the problems related with the extraction and recovery of intracellularly synthesized inorganic nanoparticles, and to devise a method to increasing the output, the shape, size, composition and dispersal of nanoparticles is not altered. Water proved to be much better system as it provided well dispersed, stable gold nanoparticles and higher recovery. This is the first report, where intracellular nanoparticles have been recovered using a very cost-effective and eco-friendly approach.

  9. 26 CFR 1.221-1 - Deduction for interest paid on qualified education loans after December 31, 2001.

    Science.gov (United States)

    2010-04-01

    ... Individuals § 1.221-1 Deduction for interest paid on qualified education loans after December 31, 2001. (a) In... interest paid by the taxpayer during the taxable year on a qualified education loan. See paragraph (b)(4... paid on qualified education loans after December 31, 2001, in taxable years ending after December 31...

  10. 26 CFR 1.221-2 - Deduction for interest due and paid on qualified education loans before January 1, 2002.

    Science.gov (United States)

    2010-04-01

    ... Individuals § 1.221-2 Deduction for interest due and paid on qualified education loans before January 1, 2002... interest due and paid by the taxpayer during the taxable year on a qualified education loan. The deduction is allowed only with respect to interest due and paid on a qualified education loan during the first...

  11. 26 CFR 1.6050S-4 - Information reporting for payments of interest on qualified education loans.

    Science.gov (United States)

    2010-04-01

    ... on qualified education loans. 1.6050S-4 Section 1.6050S-4 Internal Revenue INTERNAL REVENUE SERVICE... Information reporting for payments of interest on qualified education loans. (a) Electronic furnishing of... payments of interest on qualified education loans (furnisher) to the individual to whom it is required to...

  12. 26 CFR 1.414(r)-7 - Determination of the employees of an employer's qualified separate lines of business.

    Science.gov (United States)

    2010-04-01

    ... same department, or at the same location, to the same qualified separate line of business would be an... qualified separate lines of business. 1.414(r)-7 Section 1.414(r)-7 Internal Revenue INTERNAL REVENUE...-Sharing, Stock Bonus Plans, Etc. § 1.414(r)-7 Determination of the employees of an employer's qualified...

  13. 9 CFR 85.6 - Interstate movement of pseudorabies vaccinate swine, except swine from qualified negative gene...

    Science.gov (United States)

    2010-01-01

    ... vaccinate swine, except swine from qualified negative gene-altered vaccinated herds, not known to be..., except swine from qualified negative gene-altered vaccinated herds, not known to be infected with or exposed to pseudorabies. Pseudorabies vaccinate swine, except swine from qualified negative gene-altered...

  14. Tethering factors as organizers of intracellular vesicular traffic.

    Science.gov (United States)

    Yu, I-Mei; Hughson, Frederick M

    2010-01-01

    Intracellular trafficking entails the budding, transport, tethering, and fusion of transport vesicles and other membrane carriers. Here we review recent progress toward a mechanistic understanding of vesicle tethering. The known tethering factors are large complexes important for one or more intracellular trafficking pathways and are capable of interacting directly with many of the other principal components of the cellular trafficking machinery. Our review emphasizes recent developments in the in vitro reconstitution of vesicle tethering and the structural characterization of multisubunit tethering factors. The combination of these and other approaches has led to exciting progress toward understanding how these essential nanomachines work.

  15. Autophagic clearance of bacterial pathogens: molecular recognition of intracellular microorganisms.

    Science.gov (United States)

    Pareja, Maria Eugenia Mansilla; Colombo, Maria I

    2013-01-01

    Autophagy is involved in several physiological and pathological processes. One of the key roles of the autophagic pathway is to participate in the first line of defense against the invasion of pathogens, as part of the innate immune response. Targeting of intracellular bacteria by the autophagic machinery, either in the cytoplasm or within vacuolar compartments, helps to control bacterial proliferation in the host cell, controlling also the spreading of the infection. In this review we will describe the means used by diverse bacterial pathogens to survive intracellularly and how they are recognized by the autophagic molecular machinery, as well as the mechanisms used to avoid autophagic clearance.

  16. Space Qualified Non-Destructive Evaluation and Structural Health Monitoring Technology, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — Encouraged by Phase I accomplishments, the proposed Phase II program will significantly mature and align the development of a Space Qualified Non-Destructive...

  17. Space Qualified Non-Destructive Evaluation and Structural Health Monitoring Technology, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — NextGen Aeronautics is proposing an innovative space qualified non-destructive evaluation and health monitoring technology. The technology is built on concepts...

  18. Virginia Tech team qualifies as DARPA Urban Challenge semi-finalist

    OpenAIRE

    Crumbley, Liz

    2007-01-01

    "VictorTango," a team of Virginia Tech engineering and geography students, will travel to Victorville, Calif., for the national qualifying rounds of the Urban Challenge autonomous vehicle competition, sponsored by the Defense Advanced Research Projects Agency (DARPA).

  19. 49 CFR 604.15 - Registration of qualified human service organizations.

    Science.gov (United States)

    2010-10-01

    ... low income, advanced age, or with disabilities), shall register on FTA's charter registration Web site... service organization and whether the qualified human service organization is exempt from taxation under...

  20. 77 FR 23373 - Small Business Investment Companies-Energy Saving Qualified Investments

    Science.gov (United States)

    2012-04-19

    ... appropriate for inclusion in the definition. Furthermore, if a Small Business generates revenues solely from... definition of Energy Saving Qualified Investment by adding a presumption that a Small Business will be...

  1. Regulatory guides for qualifying the calculation methodology of Furnas by CNEN

    International Nuclear Information System (INIS)

    1987-10-01

    Regulatory guides are presented which will be used for qualifying the calculation methodology of FURNAS by CNEN, in the areas of Neutronics, Thermohydraulics, Accident Analysis and Fuel Rod Performance, as applied to Angra 1 NPP. (Author) [pt

  2. 77 FR 13009 - Regulation of Fuels and Fuel Additives: Identification of Additional Qualifying Renewable Fuel...

    Science.gov (United States)

    2012-03-05

    ... Regulation of Fuels and Fuel Additives: Identification of Additional Qualifying Renewable Fuel Pathways Under the Renewable Fuel Standard Program AGENCY: Environmental Protection Agency (EPA). ACTION: Withdrawal... Renewable Fuel Standard program regulations. Because EPA received adverse comment, we are withdrawing the...

  3. Assessment of demand for and supply of qualified manpower for the nuclear industry

    International Nuclear Information System (INIS)

    Morelle, J.

    1993-01-01

    The OECD Nuclear Energy Agency recently published a study which presents the results of a pioneering survey of the demand for and the supply of qualified manpower in various sectors of the nuclear industry (including medicine), and in the related areas of regulation and education in 12 OECD countries. The current manpower situation is presented and the future demand is reviewed. Present and future activities of OECD countries to ensure a balance between supply and demand of qualified manpower are discussed

  4. 49 CFR 40.281 - Who is qualified to act as a SAP?

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 1 2010-10-01 2010-10-01 false Who is qualified to act as a SAP? 40.281 Section... § 40.281 Who is qualified to act as a SAP? To be permitted to act as a SAP in the DOT drug and alcohol.... (2) You must be knowledgeable about the SAP function as it relates to employer interests in safety...

  5. Doctors qualifying from United Kingdom medical schools during the calendar years 1977 and 1983.

    Science.gov (United States)

    Parkhouse, J; Parkhouse, H F

    1989-01-01

    At the conclusion of undergraduate medical education in the United Kingdom most students pass a university qualifying examination and obtain a degree in medicine and surgery. Some students pass an external non-university qualifying examination in medicine as an alternative to obtaining a degree, and some do both. The degree may be obtained in the same year as the non-university qualifying examination, or in a different year. Some students from a medical school intake qualify in a later year than expected, for various reasons. Data from university, Health Department and other sources may relate to the academic year, the calendar year, or a fixed date such as 30 September. It is not a simple exercise, therefore, to define the exact number of people who qualify to practise medicine, for the first time, in any given 'year'. In counting qualifiers from individual medical schools, the problems are further compounded by the movement of students between the preclinical and clinical stages of the course, particularly from Oxford and Cambridge to London teaching hospitals. This paper analyses the situation for the calendar years 1977 and 1983, showing a decline in the number of students obtaining double (i.e. both university and non-university) qualifications. The number of UK graduates not registering with the General Medical Council to practise, at least for a time, in the UK was small, and the population base compiled for Medical Career Research Group studies was reasonably accurate in each of the 2 years examined.

  6. Essential attributes and qualifiers of primary health care

    Directory of Open Access Journals (Sweden)

    Andréa Silvia Walter de Aguiar

    2012-06-01

    variables that can assess the quality of a system or health service and rated according to their characteristics in structure, process and outcome. The evaluation of the process includes the quality of services provided by health professionals individually or in groups and refer to professional qualifications, organization and coordination of the work process of teams. The evaluation of the structure includes environmental conditions and equipment in which the services are provided and the results are evaluated starting from the verification of changes in health status of a population that can be attributed to the care process.Among the theoretical and conceptual landmarks of the PHC highlights the publication “Primary Care: Balancing Health Needs, Services, and Technology”, by Professor Barbara Starfield, in 1998, translated into Portuguese and published in Brazil in 2002. The book provides evidence on the role of PHC in health systems, evidence of its impact on population health, and compares the cost-effectiveness between countries with different forms and different degrees of implementation of this strategy, and propose a structure for measure it and set its attributes(1.The views of PHC, centered in the individual and in the population, provided the normative basis for evaluating it in a health system and contributed to the construction of the evaluation framework proposed by Starfield(1.The author also proposed a framework for evaluating the PHC which considered the concepts of the essential attributes and derivative measures of structure (capacity and process (performance.The essential and exclusive attributes of the PHC include: access / care on first contact, longitudinality, integrality and coordination of care. A high level of reach of essential attributes of the PHC results in three additional aspects, denominated derivatives, which qualify the actions and services at this level of care(1,4.The aspects qualifiers are: centered on the family

  7. The epithelial cell cytoskeleton and intracellular trafficking. I. Shiga toxin B-subunit system: retrograde transport, intracellular vectorization, and more.

    Science.gov (United States)

    Johannes, Ludger

    2002-07-01

    Many intracellular transport routes are still little explored. This is particularly true for retrograde transport between the plasma membrane and the endoplasmic reticulum. Shiga toxin B subunit has become a powerful tool to study this pathway, and recent advances on the molecular mechanisms of transport in the retrograde route and on its physiological function(s) are summarized. Furthermore, it is discussed how the study of retrograde transport of Shiga toxin B subunit allows one to design new methods for the intracellular delivery of therapeutic compounds.

  8. Monitoring intracellular oxidative events using dynamic spectral unmixing microscopy

    Science.gov (United States)

    There is increasing interest in using live-cell imaging to monitor not just individual intracellular endpoints, but to investigate the interplay between multiple molecular events as they unfold in real time within the cell. A major impediment to simultaneous acquisition of multip...

  9. FLIPR assays of intracellular calcium in GPCR drug discovery

    DEFF Research Database (Denmark)

    Hansen, Kasper Bø; Bräuner-Osborne, Hans

    2009-01-01

    Fluorescent dyes sensitive to changes in intracellular calcium have become increasingly popular in G protein-coupled receptor (GPCR) drug discovery for several reasons. First of all, the assays using the dyes are easy to perform and are of low cost compared to other assays. Second, most non...

  10. Intracellular localization of Na + /H + antiporter from Malus zumi ...

    African Journals Online (AJOL)

    In this study, we examined the intracellular localization of the product of Na+/H+ antiporter gene (MzNHX1) cloned from Malus zumi. Analysis using yeast cells expressing a fusion protein of MzNHX1 and green fluorescent protein confirmed the localization of MzNHX1 on the tonoplast.

  11. Intracellular pH gradients in migrating cells

    DEFF Research Database (Denmark)

    Martin, Christine; Pedersen, Stine Helene Falsig; Schwab, Albrecht

    2011-01-01

    might function as such unevenly distributed regulators as they modulate the interaction of focal adhesion proteins and components of the cytoskeleton in vitro. However, an intracellular pH (pH(i)) gradient reflecting a spatial asymmetry of protons has not been shown so far. One major regulator of p...

  12. Structural rearrangement of the intracellular domains during AMPA receptor activation

    DEFF Research Database (Denmark)

    Zachariassen, Linda Grønborg; Katchan, Ljudmila; Jensen, Anna Guldvang

    2016-01-01

    -clamp fluorometry of the double- and single-insert constructs showed that both the intracellular C-terminal domain (CTD) and the loop region between the M1 and M2 helices move during activation and the CTD is detached from the membrane. Our time-resolved measurements revealed unexpectedly complex fluorescence...

  13. The interferon response to intracellular DNA: why so many receptors?

    Science.gov (United States)

    Unterholzner, Leonie

    2013-11-01

    The detection of intracellular DNA has emerged to be a key event in the innate immune response to viruses and intracellular bacteria, and during conditions of sterile inflammation and autoimmunity. One of the consequences of the detection of DNA as a 'stranger' and a 'danger' signal is the production of type I interferons and pro-inflammatory cytokines. Much work has been dedicated to the elucidation of the signalling cascades that activate this DNA-induced gene expression programme. However, while many proteins have been proposed to act as sensors for intracellular DNA in recent years, none has been met with universal acceptance, and a theory linking all the recent observations is, as yet, lacking. This review presents the evidence for the various interferon-inducing DNA receptors proposed to date, and examines the hypotheses that might explain why so many different receptors appear to be involved in the innate immune recognition of intracellular DNA. Copyright © 2013 Elsevier GmbH. All rights reserved.

  14. Manipulation of Host Cholesterol by Obligate Intracellular Bacteria

    Directory of Open Access Journals (Sweden)

    Dhritiman Samanta

    2017-05-01

    Full Text Available Cholesterol is a multifunctional lipid that plays important metabolic and structural roles in the eukaryotic cell. Despite having diverse lifestyles, the obligate intracellular bacterial pathogens Chlamydia, Coxiella, Anaplasma, Ehrlichia, and Rickettsia all target cholesterol during host cell colonization as a potential source of membrane, as well as a means to manipulate host cell signaling and trafficking. To promote host cell entry, these pathogens utilize cholesterol-rich microdomains known as lipid rafts, which serve as organizational and functional platforms for host signaling pathways involved in phagocytosis. Once a pathogen gains entrance to the intracellular space, it can manipulate host cholesterol trafficking pathways to access nutrient-rich vesicles or acquire membrane components for the bacteria or bacteria-containing vacuole. To acquire cholesterol, these pathogens specifically target host cholesterol metabolism, uptake, efflux, and storage. In this review, we examine the strategies obligate intracellular bacterial pathogens employ to manipulate cholesterol during host cell colonization. Understanding how obligate intracellular pathogens target and use host cholesterol provides critical insight into the host-pathogen relationship.

  15. Engineering of obligate intracellular bacteria: progress, challenges and paradigms

    Science.gov (United States)

    Over twenty years have passed since the first report of genetic manipulation of an obligate intracellular bacterium. Through progress interspersed by bouts of stagnation, microbiologists and geneticists have developed approaches to genetically manipulate obligates. A brief overview of the current ge...

  16. Galectin-3 guides intracellular trafficking of some human serotransferrin glycoforms

    DEFF Research Database (Denmark)

    Carlsson, Carl Michael; Bengtson, Per; Cucak, Helena

    2013-01-01

    these transferrin glycoforms differently after preloading with exogenously added galectin-3. In all, this study provides the first evidence of a functional role for transferrin glycans, in intracellular trafficking after uptake. Moreover, the galectin-3 bound glycoform increased in cancer, suggesting...

  17. Dihydroceramide biology - Structure-specific metabolism and intracellular localization

    NARCIS (Netherlands)

    Kok, JW; NikolovaKarakashian, M; Klappe, K; Alexander, C; Merrill, AH

    1997-01-01

    This study utilized fluorescent analogs to characterize the intracellular transport and metabolism of dihydroceramide (DN-Cer), an intermediate in de novo sphingolipid biosynthesis, When 6-[N-(7-nitro-2,1,3-benzoxadiazol-4-yl) amino]hexanoyl-DH-Cer (C-6-NBD-DH-Cer) was incubated with HT29, NRK, BHK,

  18. CONTRIBUTIONS OF INTRACELLULAR IONS TO Kv CHANNEL VOLTAGE SENSOR DYNAMICS.

    Directory of Open Access Journals (Sweden)

    Samuel eGoodchild

    2012-06-01

    Full Text Available Voltage sensing domains of Kv channels control ionic conductance through coupling of the movement of charged residues in the S4 segment to conformational changes at the cytoplasmic region of the pore domain, that allow K+ ions to flow. Conformational transitions within the voltage sensing domain caused by changes in the applied voltage across the membrane field are coupled to the conducting pore region and the gating of ionic conductance. However, several other factors not directly linked to the voltage dependent movement of charged residues within the voltage sensor impact the dynamics of the voltage sensor, such as inactivation, ionic conductance, intracellular ion identity and block of the channel by intracellular ligands. The effect of intracellular ions on voltage sensor dynamics is of importance in the interpretation of gating current measurements and the physiology of pore/voltage sensor coupling. There is a significant amount of variability in the reported kinetics of voltage sensor deactivation kinetics of Kv channels attributed to different mechanisms such as open state stabilization, immobilization and relaxation processes of the voltage sensor. Here we separate these factors and focus on the causal role that intracellular ions can play in allosterically modulating the dynamics of Kv voltage sensor deactivation kinetics. These considerations are of critical importance in understanding the molecular determinants of the complete channel gating cycle from activation to deactivation.

  19. Biomineralization Patterns of Intracellular Carbonatogenesis in Cyanobacteria: Molecular Hypotheses

    Directory of Open Access Journals (Sweden)

    Jinhua Li

    2016-02-01

    Full Text Available The recent discovery of intracellular carbonatogenesis in several cyanobacteria species has challenged the traditional view that this process was extracellular and not controlled. However, a detailed analysis of the size distribution, chemical composition and 3-D-arrangement of carbonates in these cyanobacteria is lacking. Here, we characterized these features in Candidatus Gloeomargarita lithophora C7 and Candidatus Synechococcus calcipolaris G9 by conventional transmission electron microscopy, tomography, ultramicrotomy, and scanning transmission X-ray microscopy (STXM. Both Ca. G. lithophora C7 and Ca. S. calcipolaris G9 formed numerous polyphosphate granules adjacent or engulfing Ca-carbonate inclusions when grown in phosphate-rich solutions. Ca-carbonates were scattered within Ca. G. lithophora C7 cells under these conditions, but sometimes arranged in one or several chains. In contrast, Ca-carbonates formed at cell septa in Ca. S. calcipolaris G9 and were segregated equally between daughter cells after cell division, arranging as distorted disks at cell poles. The size distribution of carbonates evolved from a positively to a negatively skewed distribution as particles grew. Conventional ultramicrotomy did not preserve Ca-carbonates explaining partly why intracellular calcification has been overlooked in the past. All these new observations allow discussing with unprecedented insight some nucleation and growth processes occurring in intracellularly calcifying cyanobacteria with a particular emphasis on the possible involvement of intracellular compartments and cytoskeleton.

  20. Antibody- and TRIM21-dependent intracellular restriction of Salmonella enterica.

    Science.gov (United States)

    Rakebrandt, Nikolas; Lentes, Sabine; Neumann, Heinz; James, Leo C; Neumann-Staubitz, Petra

    2014-11-01

    TRIM21 ('tripartite motif-containing protein 21', Ro52) is a ubiquitously expressed cytosolic Fc receptor, which has a potent role in protective immunity against nonenveloped viruses. TRIM21 mediates intracellular neutralisation of antibody-coated viruses, a process called ADIN (antibody-dependent intracellular neutralisation). Our results reveal a similar mechanism to fight bacterial infections. TRIM21 is recruited to the intracellular pathogen Salmonella enterica in epithelial cells early in infection. TRIM21 does not bind directly to S. enterica, but to antibodies opsonising it. Most importantly, bacterial restriction is dependent on TRIM21 as well as on the opsonisation state of the bacteria. Finally, Salmonella and TRIM21 colocalise with the autophagosomal marker LC3, and intracellular defence is enhanced in starved cells suggesting an involvement of the autophagocytic pathway. Our data extend the protective role of TRIM21 from viruses to bacteria and thereby strengthening the general role of ADIN in cellular immunity. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  1. Cytoplasmic tail of coronavirus spike protein has intracellular

    Indian Academy of Sciences (India)

    https://www.ias.ac.in/article/fulltext/jbsc/042/02/0231-0244. Keywords. Coronavirus spike protein trafficking; cytoplasmic tail signal; endoplasmic reticulum–Golgi intermediate complex; lysosome. Abstract. Intracellular trafficking and localization studies of spike protein from SARS and OC43 showed that SARS spikeprotein is ...

  2. Facilitating Intracellular Drug Delivery by Ultrasound-Activated Microbubbles

    NARCIS (Netherlands)

    Lammertink, BHA

    2017-01-01

    The goal of this thesis was to investigate the combination of ultrasound and microbubbles (USMB) for intracellular delivery of (model) drugs in vitro. We have focused on clinically approved drugs, i.e. cisplatin, and microbubbles, i.e. SonoVue™, to facilitate clinical translation. In addition, model

  3. Single-cell intracellular nano-pH probes†

    Science.gov (United States)

    Özel, Rıfat Emrah; Lohith, Akshar; Mak, Wai Han; Pourmand, Nader

    2016-01-01

    Within a large clonal population, such as cancerous tumor entities, cells are not identical, and the differences between intracellular pH levels of individual cells may be important indicators of heterogeneity that could be relevant in clinical practice, especially in personalized medicine. Therefore, the detection of the intracellular pH at the single-cell level is of great importance to identify and study outlier cells. However, quantitative and real-time measurements of the intracellular pH of individual cells within a cell population is challenging with existing technologies, and there is a need to engineer new methodologies. In this paper, we discuss the use of nanopipette technology to overcome the limitations of intracellular pH measurements at the single-cell level. We have developed a nano-pH probe through physisorption of chitosan onto hydroxylated quartz nanopipettes with extremely small pore sizes (~100 nm). The dynamic pH range of the nano-pH probe was from 2.6 to 10.7 with a sensitivity of 0.09 units. We have performed single-cell intracellular pH measurements using non-cancerous and cancerous cell lines, including human fibroblasts, HeLa, MDA-MB-231 and MCF-7, with the pH nanoprobe. We have further demonstrated the real-time continuous single-cell pH measurement capability of the sensor, showing the cellular pH response to pharmaceutical manipulations. These findings suggest that the chitosan-functionalized nanopore is a powerful nano-tool for pH sensing at the single-cell level with high temporal and spatial resolution. PMID:27708772

  4. Single-cell intracellular nano-pH probes.

    Science.gov (United States)

    Özel, Rıfat Emrah; Lohith, Akshar; Mak, Wai Han; Pourmand, Nader

    2015-01-01

    Within a large clonal population, such as cancerous tumor entities, cells are not identical, and the differences between intracellular pH levels of individual cells may be important indicators of heterogeneity that could be relevant in clinical practice, especially in personalized medicine. Therefore, the detection of the intracellular pH at the single-cell level is of great importance to identify and study outlier cells. However, quantitative and real-time measurements of the intracellular pH of individual cells within a cell population is challenging with existing technologies, and there is a need to engineer new methodologies. In this paper, we discuss the use of nanopipette technology to overcome the limitations of intracellular pH measurements at the single-cell level. We have developed a nano-pH probe through physisorption of chitosan onto hydroxylated quartz nanopipettes with extremely small pore sizes (~100 nm). The dynamic pH range of the nano-pH probe was from 2.6 to 10.7 with a sensitivity of 0.09 units. We have performed single-cell intracellular pH measurements using non-cancerous and cancerous cell lines, including human fibroblasts, HeLa, MDA-MB-231 and MCF-7, with the pH nanoprobe. We have further demonstrated the real-time continuous single-cell pH measurement capability of the sensor, showing the cellular pH response to pharmaceutical manipulations. These findings suggest that the chitosan-functionalized nanopore is a powerful nano-tool for pH sensing at the single-cell level with high temporal and spatial resolution.

  5. Legionella pneumophila transcriptome during intracellular multiplication in human macrophages

    Directory of Open Access Journals (Sweden)

    Sebastien P Faucher

    2011-04-01

    Full Text Available Legionella pneumophila is the causative agent of Legionnaires’ disease, an acute pulmonary infection. L. pneumophila is able to infect and multiply in both phagocytic protozoa, such as Acanthamoeba castellanii, and mammalian professional phagocytes. The best-known L. pneumophila virulence determinant is the Icm/Dot Type IVB secretion system (TFBSS, which is used to translocate more than 150 effector proteins to host cells. While the transcriptional response of Legionella to the intracellular environment of A. castellanii has been investigated, much less is known about the Legionella transcriptional response inside human macrophages. In this study, the transcriptome of L. pneumophila was monitored during exponential and post-exponential phase in rich AYE broth as well as during infection of human cultured macrophages. This was accomplished with microarrays and an RNA amplification procedure called SCOTS to detect small amounts of mRNA from low numbers of intracellular bacteria. Among the genes induced intracellularly are those involved in amino acid biosynthetic pathways leading to L-arginine, L-histidine and L-proline as well as many transport systems involved in amino acid and iron uptake. Gene involved in catabolism of glycerol is also induced during intracellular growth and could be used as a carbon source. The genes encoding the Icm/Dot system are not differentially expressed inside cells compared to control bacteria grown in rich broth, but the genes encoding several translocated effectors are strongly induced. Moreover, we used the transcriptome data to predict previously unrecognized Icm/Dot effector genes based on their expression pattern and confirmed translocation for three candidates. This study provides a comprehensive view of how L. pneumophila responds to the human macrophage intracellular environment.

  6. Microsporidia are natural intracellular parasites of the nematode Caenorhabditis elegans.

    Science.gov (United States)

    Troemel, Emily R; Félix, Marie-Anne; Whiteman, Noah K; Barrière, Antoine; Ausubel, Frederick M

    2008-12-09

    For decades the soil nematode Caenorhabditis elegans has been an important model system for biology, but little is known about its natural ecology. Recently, C. elegans has become the focus of studies of innate immunity and several pathogens have been shown to cause lethal intestinal infections in C. elegans. However none of these pathogens has been shown to invade nematode intestinal cells, and no pathogen has been isolated from wild-caught C. elegans. Here we describe an intracellular pathogen isolated from wild-caught C. elegans that we show is a new species of microsporidia. Microsporidia comprise a large class of eukaryotic intracellular parasites that are medically and agriculturally important, but poorly understood. We show that microsporidian infection of the C. elegans intestine proceeds through distinct stages and is transmitted horizontally. Disruption of a conserved cytoskeletal structure in the intestine called the terminal web correlates with the release of microsporidian spores from infected cells, and appears to be part of a novel mechanism by which intracellular pathogens exit from infected cells. Unlike in bacterial intestinal infections, the p38 MAPK and insulin/insulin-like growth factor (IGF) signaling pathways do not appear to play substantial roles in resistance to microsporidian infection in C. elegans. We found microsporidia in multiple wild-caught isolates of Caenorhabditis nematodes from diverse geographic locations. These results indicate that microsporidia are common parasites of C. elegans in the wild. In addition, the interaction between C. elegans and its natural microsporidian parasites provides a system in which to dissect intracellular intestinal infection in vivo and insight into the diversity of pathogenic mechanisms used by intracellular microbes.

  7. Experience in initial training required for the recognition of the qualified RP expert in Spain

    International Nuclear Information System (INIS)

    Rodriguez Suarez, M.; Marco Arboli, M.; Menarguez, J.

    2003-01-01

    An important point of the actions inside the European framework to achieve the harmonisation of the training programmes and recognition was included in the European directive 96/29/Euratom which includes definition and specific tasks of the European Qualified Expert on Radiation Protection (RP). Basic syllabus for training of those experts was developed in the communication 98/C 133/03 concerning BSS applications. Although, in the Spanish education system, the training and recognition requirements of the high level qualified experts on RP was defined since 1977, until 2001, the figure of the Technical Qualified Expert on RP does not appear in the legal framework. In December 2002, a new regulation of the Spanish Regulatory Body, CSN, about qualifications to obtain the recognition of RP Expert in Spain (both high qualified and technical RP experts) was published. Concerning the qualified expert on RP, (RP Officer), responsible of the RP Service, which takes charge of the effective protection and advise radioactive and nuclear facilities in Rp aspects,has to be authorised by the regulatory body. to obtain the RP officer diploma, conceded by the CSN, an initial training of 300 hours and a three-year minimum experience are required (for X-ray installation a 6-month experience is enough). The technical qualified expert on RP is the worker who carried out the tasks in the a RP Service under the supervision of the RP officer. A Technician Qualified Expert on RP does not need an specific accreditation of the Regulatory Body, but an initial RP training and a three-month minimum experience are required and has hold a certificate by the RP officer. Continuous training is also required and as well has to receive a certificate from the RP officer. Since 1977, The Institute for energy Studies has been implementing specific training courses for those professionals who want to obtain the diploma of RP officer (high degree qualified RP expert), conceded by the CSN. Since then

  8. Enhanced intracellular delivery and antibacterial efficacy of enrofloxacin-loaded docosanoic acid solid lipid nanoparticles against intracellular Salmonella.

    Science.gov (United States)

    Xie, Shuyu; Yang, Fei; Tao, Yanfei; Chen, Dongmei; Qu, Wei; Huang, Lingli; Liu, Zhenli; Pan, Yuanhu; Yuan, Zonghui

    2017-01-23

    Enrofloxacin-loaded docosanoic acid solid lipid nanoparticles (SLNs) with different physicochemical properties were developed to enhance activity against intracellular Salmonella. Their cellular uptake, intracellular elimination and antibacterial activity were studied in RAW 264.7 cells. During the experimental period, SLN-encapsulated enrofloxacin accumulated in the cells approximately 27.06-37.71 times more efficiently than free drugs at the same extracellular concentration. After incubation for 0.5 h, the intracellular enrofloxacin was enhanced from 0.336 to 1.147 μg/mg of protein as the sizes of nanoparticles were increased from 150 to 605 nm, and from 0.960 to 1.147 μg/mg of protein when the charge was improved from -8.1 to -24.9 mv. The cellular uptake was more significantly influenced by the size than it was by the charge, and was not affected by whether the charge was positive or negative. The elimination of optimal SLN-encapsulated enrofloxacin from the cells was significantly slower than that of free enrofloxacin after removing extracellular drug. The inhibition effect against intracellular Salmonella CVCC541 of 0.24 and 0.06 μg/mL encapsulated enrofloxacin was stronger than 0.6 μg/mL free drug after all of the incubation periods and at 48 h, respectively. Docosanoic acid SLNs are thus considered as a promising carrier for intracellular bacterial treatment.

  9. Using neutrosophic graph cut segmentation algorithm for qualified rendering image selection in thyroid elastography video.

    Science.gov (United States)

    Guo, Yanhui; Jiang, Shuang-Quan; Sun, Baiqing; Siuly, Siuly; Şengür, Abdulkadir; Tian, Jia-Wei

    2017-12-01

    Recently, elastography has become very popular in clinical investigation for thyroid cancer detection and diagnosis. In elastogram, the stress results of the thyroid are displayed using pseudo colors. Due to variation of the rendering results in different frames, it is difficult for radiologists to manually select the qualified frame image quickly and efficiently. The purpose of this study is to find the qualified rendering result in the thyroid elastogram. This paper employs an efficient thyroid ultrasound image segmentation algorithm based on neutrosophic graph cut to find the qualified rendering images. Firstly, a thyroid ultrasound image is mapped into neutrosophic set, and an indeterminacy filter is constructed to reduce the indeterminacy of the spatial and intensity information in the image. A graph is defined on the image and the weight for each pixel is represented using the value after indeterminacy filtering. The segmentation results are obtained using a maximum-flow algorithm on the graph. Then the anatomic structure is identified in thyroid ultrasound image. Finally the rendering colors on these anatomic regions are extracted and validated to find the frames which satisfy the selection criteria. To test the performance of the proposed method, a thyroid elastogram dataset is built and totally 33 cases were collected. An experienced radiologist manually evaluates the selection results of the proposed method. Experimental results demonstrate that the proposed method finds the qualified rendering frame with 100% accuracy. The proposed scheme assists the radiologists to diagnose the thyroid diseases using the qualified rendering images.

  10. Regulation of dopamine transporter trafficking by intracellular amphetamine

    DEFF Research Database (Denmark)

    Kahlig, Kristopher M; Lute, Brandon J; Wei, Yuqiang

    2006-01-01

    -induced cell surface DAT redistribution may result in long-lasting changes in DA homeostasis. The molecular mechanism by which AMPH induces trafficking is not clear. Because AMPH is a substrate, we do not know whether extracellular AMPH stimulates trafficking through its interaction with DAT and subsequent...... alteration in DAT function, thereby triggering intracellular signaling or whether AMPH must be transported and then act intracellularly. In agreement with our previous studies, extracellular AMPH caused cytosolic redistribution of the wild-type human DAT (WT-hDAT). However, AMPH did not induce cytosolic...... redistribution in an uptake-impaired hDAT (Y335A-hDAT) that still binds AMPH. The divalent cation zinc (Zn(2+)) inhibits WT-hDAT activity, but it restores Y335A-hDAT uptake. Coadministration of Zn(2+) and AMPH consistently reduced WT-hDAT trafficking but stimulated cytosolic redistribution of Y335A...

  11. Pico gauges for minimally invasive intracellular hydrostatic pressure measurements

    DEFF Research Database (Denmark)

    Knoblauch, Jan; Mullendore, Daniel L.; Jensen, Kaare Hartvig

    2014-01-01

    Intracellular pressure has a multitude of functions in cells surrounded by a cell wall or similar matrix in all kingdoms of life. The functions include cell growth, nastic movements, and penetration of tissue by parasites. The precise measurement of intracellular pressure in the majority of cells......, however, remains difficult or impossible due to their small size and/or sensitivity to manipulation. Here, we report on a method that allows precise measurements in basically any cell type over all ranges of pressure. It is based on the compression of nanoliter and picoliter volumes of oil entrapped...... in the tip of microcapillaries, which we call pico gauges. The production of pico gauges can be accomplished with standard laboratory equipment, and measurements are comparably easy to conduct. Example pressure measurements are performed on cells that are difficult or impossible to measure with other methods....

  12. Intracellular compartmentalization of skeletal muscle glycogen metabolism and insulin signalling

    DEFF Research Database (Denmark)

    Prats Gavalda, Clara; Gomez-Cabello, Alba; Vigelsø Hansen, Andreas

    2011-01-01

    The interest in skeletal muscle metabolism and insulin signalling has increased exponentially in recent years as a consequence of their role in the development of type 2 diabetes mellitus. Despite this, the exact mechanisms involved in the regulation of skeletal muscle glycogen metabolism...... and insulin signalling transduction remain elusive. We believe that one of the reasons is that the role of intracellular compartmentalization as a regulator of metabolic pathways and signalling transduction has been rather ignored. This paper briefly reviews the literature to discuss the role of intracellular...... compartmentalization in the regulation of skeletal muscle glycogen metabolism and insulin signalling. As a result, a hypothetical regulatory mechanism is proposed by which cells could direct glycogen resynthesis towards different pools of glycogen particles depending on the metabolic needs. Furthermore, we discuss...

  13. Intracellular Chemistry: Integrating Molecular Inorganic Catalysts with Living Systems.

    Science.gov (United States)

    Ngo, Anh H; Bose, Sohini; Do, Loi H

    2018-03-23

    This concept article focuses on the rapid growth of intracellular chemistry dedicated to the integration of small-molecule metal catalysts with living cells and organisms. Although biological systems contain a plethora of biomolecules that can deactivate inorganic species, researchers have shown that small-molecule metal catalysts could be engineered to operate in heterogeneous aqueous environments. Synthetic intracellular reactions have recently been reported for olefin hydrogenation, hydrolysis/oxidative cleavage, azide-alkyne cycloaddition, allylcarbamate cleavage, C-C bond cross coupling, and transfer hydrogenation. Other promising targets for new biocompatible reaction discovery will also be discussed, with a special emphasis on how such innovations could lead to the development of novel technologies and chemical tools. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Chelation of intracellular calcium blocks insulin action in the adipocyte

    International Nuclear Information System (INIS)

    Pershadsingh, H.A.; Shade, D.L.; Delfert, D.M.; McDonald, J.M.

    1987-01-01

    The hypothesis that intracellular Ca 2+ is an essential component of the intracellular mechanism of insulin action in the adipocyte was evaluated. Cells were loaded with the Ca 2+ chelator quin-2, by preincubating them with quin-2 AM, the tetrakis(acetoxymethyl) ester of quin-2. Quin-2 loading inhibited insulin-stimulated glucose transport without affecting basal activity. The ability of insulin to stimulate glucose uptake in quin-2-loaded cells could be partially restored by preincubating cells with buffer supplemented with 1.2 mM CaCl 2 and the Ca 2+ ionophore A23187. These conditions had no effect on basal activity and omission of CaCl 2 from the buffer prevented the restoration of insulin-stimulated glucose uptake by A23187. Quin-2 loading also inhibited insulin-stimulated glucose oxidation and the ability of insulin to inhibit cAMP-stimulated lipolysis without affecting their basal activities. Incubation of cells with 100 μM quin-2 or quin-2 AM had no effect on intracellular ATP concentration or the specific binding of 125 I=labeled insulin to adipocytes. These findings suggest that intracellular Ca 2+ is an essential component in the coupling of the insulin-activated receptor complex to cellular physiological/metabolic machinery. Furthermore, differing quin-2 AM dose-response profiles suggest the presence of dual Ca 2+ -dependent pathways in the adipocyte. One involves insulin stimulation of glucose transport and oxidation, whereas the other involves the antilipolytic action of insulin

  15. Intracellular Detection of Viral Transcription and Replication Using RNA FISH

    Science.gov (United States)

    2016-05-26

    Chapter 14. Intracellular detection of viral transcription and replication using RNA FISH i. Summary/Abstract Many hemorrhagic fever viruses...only allow entirely new investigations into the replication of these viruses, but also how this method can be applied to any virus with a known...localization, TurboFISH, hemorrhagic fever virus replication 1. Introduction RNA FISH was developed as a method to visualize cellular RNA by binding a

  16. Mycobacterium avium-intracellulare: a rare cause of subacromial bursitis.

    Science.gov (United States)

    Sinha, Raj; Tuckett, John; Hide, Geoff; Dildey, Petra; Karsandas, Alvin

    2015-01-01

    Septic subacromial bursitis is an uncommon disorder with only a few reported cases in the literature. The most common causative organism is Staphylococcus aureus. We report the case of a 61-year-old female with a septic subacromial bursitis where the causative organism was found to be Mycobacterium avium-intracellulare (MAI). The diagnosis was only made following a biopsy, and we use this case to highlight the importance of recognising the need to consider a biopsy and aspiration in atypical situations.

  17. Bullous pemphigoid antigen localization suggests an intracellular association with hemidesmosomes

    DEFF Research Database (Denmark)

    Westgate, G E; Weaver, A C; Couchman, J R

    1985-01-01

    immunofluorescent staining for BPA is linear at the basement membrane zone (BMZ) of skin and many other epithelial tissues. At higher magnification however, we observed a punctate staining pattern for BPA which was regular in appearance and suggested localization of BPA to discrete structures at the BMZ. Subsequent...... intracellularly both in vivo and in vitro. We suggest that BPA is not normally a lamina lucida component, but that it may form part of a linkage between the cytoskeleton and the basement membrane....

  18. NAD+-Glycohydrolase Promotes Intracellular Survival of Group A Streptococcus.

    Directory of Open Access Journals (Sweden)

    Onkar Sharma

    2016-03-01

    Full Text Available A global increase in invasive infections due to group A Streptococcus (S. pyogenes or GAS has been observed since the 1980s, associated with emergence of a clonal group of strains of the M1T1 serotype. Among other virulence attributes, the M1T1 clone secretes NAD+-glycohydrolase (NADase. When GAS binds to epithelial cells in vitro, NADase is translocated into the cytosol in a process mediated by streptolysin O (SLO, and expression of these two toxins is associated with enhanced GAS intracellular survival. Because SLO is required for NADase translocation, it has been difficult to distinguish pathogenic effects of NADase from those of SLO. To resolve the effects of the two proteins, we made use of anthrax toxin as an alternative means to deliver NADase to host cells, independently of SLO. We developed a novel method for purification of enzymatically active NADase fused to an amino-terminal fragment of anthrax toxin lethal factor (LFn-NADase that exploits the avid, reversible binding of NADase to its endogenous inhibitor. LFn-NADase was translocated across a synthetic lipid bilayer in vitro in the presence of anthrax toxin protective antigen in a pH-dependent manner. Exposure of human oropharyngeal keratinocytes to LFn-NADase in the presence of protective antigen resulted in cytosolic delivery of NADase activity, inhibition of protein synthesis, and cell death, whereas a similar construct of an enzymatically inactive point mutant had no effect. Anthrax toxin-mediated delivery of NADase in an amount comparable to that observed during in vitro infection with live GAS rescued the defective intracellular survival of NADase-deficient GAS and increased the survival of SLO-deficient GAS. Confocal microscopy demonstrated that delivery of LFn-NADase prevented intracellular trafficking of NADase-deficient GAS to lysosomes. We conclude that NADase mediates cytotoxicity and promotes intracellular survival of GAS in host cells.

  19. Molecular evolution, intracellular organization, and the quinary structure of proteins.

    OpenAIRE

    McConkey, E H

    1982-01-01

    High-resolution two-dimensional polyacrylamide gel electrophoresis shows that at least half of 370 denatured polypeptides from hamster cells and human cells are indistinguishable in terms of isoelectric points and molecular weights. Molecular evolution may have been more conservative for this set of proteins than sequence studies on soluble proteins have implied. This may be a consequence of complexities of intracellular organization and the numerous macromolecular interactions in which most ...

  20. Control of intracellular heme levels: Heme transporters and heme oxygenases

    OpenAIRE

    Khan, Anwar A.; Quigley, John G.

    2011-01-01

    Heme serves as a co-factor in proteins involved in fundamental biological processes including oxidative metabolism, oxygen storage and transport, signal transduction and drug metabolism. In addition, heme is important for systemic iron homeostasis in mammals. Heme has important regulatory roles in cell biology, yet excessive levels of intracellular heme are toxic; thus, mechanisms have evolved to control the acquisition, synthesis, catabolism and expulsion of cellular heme. Recently, a number...

  1. Evaluation of Intracellular Signaling Downstream Chimeric Antigen Receptors.

    Directory of Open Access Journals (Sweden)

    Hannah Karlsson

    Full Text Available CD19-targeting CAR T cells have shown potency in clinical trials targeting B cell leukemia. Although mainly second generation (2G CARs carrying CD28 or 4-1BB have been investigated in patients, preclinical studies suggest that third generation (3G CARs with both CD28 and 4-1BB have enhanced capacity. However, little is known about the intracellular signaling pathways downstream of CARs. In the present work, we have analyzed the signaling capacity post antigen stimulation in both 2G and 3G CARs. 3G CAR T cells expanded better than 2G CAR T cells upon repeated stimulation with IL-2 and autologous B cells. An antigen-driven accumulation of CAR+ cells was evident post antigen stimulation. The cytotoxicity of both 2G and 3G CAR T cells was maintained by repeated stimulation. The phosphorylation status of intracellular signaling proteins post antigen stimulation showed that 3G CAR T cells had a higher activation status than 2G. Several proteins involved in signaling downstream the TCR were activated, as were proteins involved in the cell cycle, cell adhesion and exocytosis. In conclusion, 3G CAR T cells had a higher degree of intracellular signaling activity than 2G CARs which may explain the increased proliferative capacity seen in 3G CAR T cells. The study also indicates that there may be other signaling pathways to consider when designing or evaluating new generations of CARs.

  2. Fatty Acid Signaling: The New Function of Intracellular Lipases

    Directory of Open Access Journals (Sweden)

    Zuzana Papackova

    2015-02-01

    Full Text Available Until recently, intracellular triacylglycerols (TAG stored in the form of cytoplasmic lipid droplets have been considered to be only passive “energy conserves”. Nevertheless, degradation of TAG gives rise to a pleiotropic spectrum of bioactive intermediates, which may function as potent co-factors of transcription factors or enzymes and contribute to the regulation of numerous cellular processes. From this point of view, the process of lipolysis not only provides energy-rich equivalents but also acquires a new regulatory function. In this review, we will concentrate on the role that fatty acids liberated from intracellular TAG stores play as signaling molecules. The first part provides an overview of the transcription factors, which are regulated by fatty acids derived from intracellular stores. The second part is devoted to the role of fatty acid signaling in different organs/tissues. The specific contribution of free fatty acids released by particular lipases, hormone-sensitive lipase, adipose triacylglycerol lipase and lysosomal lipase will also be discussed.

  3. Estimating the biophysical properties of neurons with intracellular calcium dynamics.

    Science.gov (United States)

    Ye, Jingxin; Rozdeba, Paul J; Morone, Uriel I; Daou, Arij; Abarbanel, Henry D I

    2014-06-01

    We investigate the dynamics of a conductance-based neuron model coupled to a model of intracellular calcium uptake and release by the endoplasmic reticulum. The intracellular calcium dynamics occur on a time scale that is orders of magnitude slower than voltage spiking behavior. Coupling these mechanisms sets the stage for the appearance of chaotic dynamics, which we observe within certain ranges of model parameter values. We then explore the question of whether one can, using observed voltage data alone, estimate the states and parameters of the voltage plus calcium (V+Ca) dynamics model. We find the answer is negative. Indeed, we show that voltage plus another observed quantity must be known to allow the estimation to be accurate. We show that observing both the voltage time course V(t) and the intracellular Ca time course will permit accurate estimation, and from the estimated model state, accurate prediction after observations are completed. This sets the stage for how one will be able to use a more detailed model of V+Ca dynamics in neuron activity in the analysis of experimental data on individual neurons as well as functional networks in which the nodes (neurons) have these biophysical properties.

  4. Extracellular and Intracellular Mechanisms Mediating Metastatic Activity of Exogenous Osteopontin

    Science.gov (United States)

    Mandelin, Jami; Lin, Emme C. K.; Hu, Dana D.; Knowles, Susan K.; Do, Kim-Anh; Wang, Xuemei; Sage, E. Helene; Smith, Jeffrey W.; Arap, Wadih; Pasqualini, Renata

    2009-01-01

    BACKGROUND Osteopontin affects several steps of the metastatic cascade. Despite direct correlation with metastasis in experimental systems and in patient studies, the extracellular and intracellular basis for these observations remains unsolved. We used human melanoma and sarcoma cell lines to evaluate the effects of soluble osteopontin on metastasis. METHODS Exogenous osteopontin or negative controls, including a site-directed mutant osteopontin, were used in functional assays in vitro, ex vivo, and in vivo designed to test extracellular and intracellular mechanisms involved in experimental metastasis. RESULTS In the extracellular environment, we confirm that soluble osteopontin is required for its pro-metastatic effects; this phenomenon is specific, RGD-dependent, and evident in experimental models of metastasis. In the intracellular environment, osteopontin initially induces rapid Tyr-418 dephosphorylation of c-Src, with decreases in actin stress fibers and increased binding to the vascular endothelium. This heretofore undescribed Tyr dephosphorylation is followed by a tandem c-Src phosphorylation after tumor cell attachment to the metastatic site. CONCLUSION Our results reveal a complex molecular interaction as well as a dual role for osteopontin in metastasis that is dependent on whether tumor cells are in circulation or attached. Such context-dependent functional insights may contribute to anti-metastasis strategies. PMID:19224553

  5. New intracellular activities of matrix metalloproteinases shine in the moonlight.

    Science.gov (United States)

    Jobin, Parker G; Butler, Georgina S; Overall, Christopher M

    2017-11-01

    Adaption of a single protein to perform multiple independent functions facilitates functional plasticity of the proteome allowing a limited number of protein-coding genes to perform a multitude of cellular processes. Multifunctionality is achievable by post-translational modifications and by modulating subcellular localization. Matrix metalloproteinases (MMPs), classically viewed as degraders of the extracellular matrix (ECM) responsible for matrix protein turnover, are more recently recognized as regulators of a range of extracellular bioactive molecules including chemokines, cytokines, and their binders. However, growing evidence has convincingly identified select MMPs in intracellular compartments with unexpected physiological and pathological roles. Intracellular MMPs have both proteolytic and non-proteolytic functions, including signal transduction and transcription factor activity thereby challenging their traditional designation as extracellular proteases. This review highlights current knowledge of subcellular location and activity of these "moonlighting" MMPs. Intracellular roles herald a new era of MMP research, rejuvenating interest in targeting these proteases in therapeutic strategies. This article is part of a Special Issue entitled: Matrix Metalloproteinases edited by Rafael Fridman. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Intracellular Hg(0) Oxidation in Desulfovibrio desulfuricans ND132.

    Science.gov (United States)

    Wang, Yuwei; Schaefer, Jeffra K; Mishra, Bhoopesh; Yee, Nathan

    2016-10-03

    The disposal of elemental mercury (Hg(0)) wastes in mining and manufacturing areas has caused serious soil and groundwater contamination issues. Under anoxic conditions, certain anaerobic bacteria can oxidize dissolved elemental mercury and convert the oxidized Hg to neurotoxic methylmercury. In this study, we conducted experiments with the Hg-methylating bacterium Desulfovibrio desulfuricans ND132 to elucidate the role of cellular thiols in anaerobic Hg(0) oxidation. The concentrations of cell-surface and intracellular thiols were measured, and specific fractions of D. desulfuricans ND132 were examined for Hg(0) oxidation activity and analyzed with extended X-ray absorption fine structure (EXAFS) spectroscopy. The experimental data indicate that intracellular thiol concentrations are approximately six times higher than those of the cell wall. Cells reacted with a thiol-blocking reagent were severely impaired in Hg(0) oxidation activity. Spheroplasts lacking cell walls rapidly oxidized Hg(0) to Hg(II), while cell wall fragments exhibited low reactivity toward Hg(0). EXAFS analysis of spheroplast samples revealed that multiple different forms of Hg-thiols are produced by the Hg(0) oxidation reaction and that the local coordination environment of the oxidized Hg changes with reaction time. The results of this study indicate that Hg(0) oxidation in D. desulfuricans ND132 is an intracellular process that occurs by reaction with thiol-containing molecules.

  7. A longitudinal, mixed methods investigation of newly qualified nurses' workplace stressors and stress experiences during transition.

    Science.gov (United States)

    Halpin, Yvonne; Terry, Louise M; Curzio, Joan

    2017-11-01

    To investigate transition in newly qualified nurses through an exploration of their stressors and stress experiences during their first 12 months postqualifying. Globally, thousands of new nurses qualify annually. They are crucial for the profession and healthcare service delivery. Work-related stress has multiple serious consequences, yet there is a lack of robust, empirical evidence that directly analyses newly qualified nurses and the stress they feel and experience in the workplace. Understanding what causes newly qualified nurses' stress is vital to retaining and nurturing this vital component of the workforce. Longitudinal, explanatory sequential mixed methods, cohort study. At the point of qualification (n = 288), 6 months postqualifying (n = 107) and 12 months postqualifying (n = 86), newly qualified nurses completed the Nursing Stress Scale, with 14 completing a one-to-one interview at 12 months postqualifying. Data were collected from 2010 - 2012. Inferential statistics, "thematic analysis" and "side-by-side comparisons in a discussion" were used for analysis. Workload was consistently the highest reported stressor with inadequate staffing and managing multiple role demands given as explanations. Incivility within the workplace was a noted stressor. Conversely, being part of "a good team" provided a civil, supportive, facilitative work environment. Entering nurse education with previous healthcare experience had a mediating effect on the reported frequency of stressors. Newly qualified nurses encounter multiple work-related stressors over their first 12 months postqualifying, which are intrinsically entwined with their transition. Employing organizations need to be more proactive in managing their workload and addressing workplace incivility. © 2017 John Wiley & Sons Ltd.

  8. Nursing educator perspectives of overseas qualified nurses' intercultural clinical communication: barriers, enablers and engagement strategies.

    Science.gov (United States)

    Philip, Susan; Manias, Elizabeth; Woodward-Kron, Robyn

    2015-09-01

    To understand the intercultural communication experiences and associated communication training needs of overseas qualified nurses in the Australian healthcare system from the unique perspectives of nurse educators teaching in accredited bridging programmes. Overseas qualified nurses are an integral part of the nursing workforce in migration destination countries. Communication training needs are more complex when there are cultural, ethnic and language differences between nurses, other health professionals and patients. A qualitative, exploratory research design using semi-structured interviews. All (nine) organisations involved in conducting the Australian Health Practitioner Regulation Agency approved preregistration bridging programmes for overseas qualified nurses within the state of Victoria, Australia, were involved in the study. Participants were 12 nurse educators employed in these organisations. Thematic analysis was undertaken. Three macro themes emerged about the overseas qualified nurses' intercultural communication: (1) pre-existing barriers and enablers to intercultural communication, for example, nurses' reluctance to engage in communicative strategies that build rapport with patients, (2) transitional behaviours and impact on communication, including maintenance of perceived cultural hierarchies between health professionals and (3) development of communicative competence, including expanding one's repertoire of conversational gambits. The findings point to the domains and causes of communication challenges facing overseas qualified nurses in new healthcare settings as well as strategies that the nurse educators and nurses can adopt. Communication cannot be merely regarded as a skill that can be taught in a didactic programme. Comprehensive understanding is needed about the sociocultural dimensions of these nurses' orientation, which can impact on how they communicate in their new healthcare settings. The findings can act as triggers for discussion

  9. Intracellular diffusion restrictions in isolated cardiomyocytes from rainbow trout

    Directory of Open Access Journals (Sweden)

    Birkedal Rikke

    2009-12-01

    Full Text Available Abstract Background Restriction of intracellular diffusion of adenine nucleotides has been studied intensively on adult rat cardiomyocytes. However, their cause and role in vivo is still uncertain. Intracellular membrane structures have been suggested to play a role. We therefore chose to study cardiomyocytes from rainbow trout (Oncorhynchus mykiss, which are thinner and have fewer intracellular membrane structures than adult rat cardiomyocytes. Previous studies suggest that trout permeabilized cardiac fibers also have diffusion restrictions. However, results from fibers may be affected by incomplete separation of the cells. This is avoided when studying permeabilized, isolated cardiomyocytes. The aim of this study was to verify the existence of diffusion restrictions in trout cardiomyocytes by comparing ADP-kinetics of mitochondrial respiration in permeabilized fibers, permeabilized cardiomyocytes and isolated mitochondria from rainbow trout heart. Experiments were performed at 10, 15 and 20°C in the absence and presence of creatine. Results Trout cardiomyocytes hypercontracted in the solutions used for mammalian cardiomyocytes. We developed a new solution in which they retained their shape and showed stable steady state respiration rates throughout an experiment. The apparent ADP-affinity of permeabilized cardiomyocytes was different from that of fibers. It was higher, independent of temperature and not increased by creatine. However, it was still about ten times lower than in isolated mitochondria. Conclusions The differences between fibers and cardiomyocytes suggest that results from trout heart fibers were affected by incomplete separation of the cells. However, the lower ADP-affinity of cardiomyocytes compared to isolated mitochondria indicate that intracellular diffusion restrictions are still present in trout cardiomyocytes despite their lower density of intracellular membrane structures. The lack of a creatine effect indicates that

  10. Roles and Responsibilities, and Education and Training Requirements for Clinically Qualified Medical Physicists (Russian Edition)

    International Nuclear Information System (INIS)

    2014-01-01

    This publication addresses the shortfall of well trained and clinically qualified medical physicists working in radiation medicine. The roles, responsibilities and clinical training requirements of medical physicists have not always been well defined or well understood by health care professionals, health authorities and regulatory agencies. To fill this gap, this publication provides recommendations for the academic education and clinical training of clinically qualified medical physicists, including recommendations for their accreditation certification and registration, along with continuous professional development. The goal is to establish criteria that support the harmonization of education and clinical training worldwide

  11. Target language use in Modern Language classrooms: perception and change among newly qualified teachers in Scotland

    OpenAIRE

    Lynch, Michael Patrick

    2015-01-01

    In this thesis I investigate the practices and perceptions of some Newly Qualified Teachers (NQTs) of modern foreign languages (MFL) in Scotland in relation to how they use the target language (L2). I seek to answer the questions “In what different ways do student teachers of modern languages use the target language in Scottish secondary school classrooms?’, ‘What reasons do they give for how they use it?” and “In what way(s), if any, do newly qualified teachers of modern language...

  12. Going beyond "highly-qualified personnel" to "value-added personnel"

    Science.gov (United States)

    Nantel, Marc; Fox, Bradley; Gelsthorpe, Leanne; Hamilton, Roxy; Marshall, Robert; Williamson, Guida

    2005-10-01

    Employers clamour for more "Highly-Qualified Personnel" (HQP) to hire, and photonics is no exception. The challenge facing all new graduates lies in what actually constitutes a Highly-Qualified Person. Yesterday, an HQP was often understood to be a graduate with a Bachelor's, Master's or Ph.D. degree. Today, industry is demanding that an HQP be either a university or college graduate with strong academic standing AND a sound business and communications skill set. In this paper, we introduce the concept of "Value-Added Personnel" (VAP), which marries both scientific and "soft-skills" aspects, and we present a new program in Ontario to produce VAPs.

  13. DMPD: Intracellular DNA sensors in immunity. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 18573338 Intracellular DNA sensors in immunity. Takeshita F, Ishii KJ. Curr Opin Im...munol. 2008 Aug;20(4):383-8. Epub 2008 Jun 23. (.png) (.svg) (.html) (.csml) Show Intracellular DNA sensors ...in immunity. PubmedID 18573338 Title Intracellular DNA sensors in immunity. Authors Takeshita F, Ishii KJ. P

  14. DMPD: NOD-like receptors (NLRs): bona fide intracellular microbial sensors. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 18585455 NOD-like receptors (NLRs): bona fide intracellular microbial sensors. Shaw...tml) (.csml) Show NOD-like receptors (NLRs): bona fide intracellular microbial sensors. PubmedID 18585455 Ti...tle NOD-like receptors (NLRs): bona fide intracellular microbial sensors. Authors

  15. 78 FR 35155 - Establishing a List of Qualifying Pathogens Under the Food and Drug Administration Safety and...

    Science.gov (United States)

    2013-06-12

    ... is thought to be a chromosomally encoded mutation in the target enzyme, although alternative... of qualifying pathogens. E. Candida Species Candida species are fungi (specifically, yeast) that are...

  16. 26 CFR 26.2652-2 - Special election for qualified terminable interest property.

    Science.gov (United States)

    2010-04-01

    ... executor) may elect to treat the trust as two separate trusts, one of which has a zero inclusion ratio by... leaving $4 million in trust for the benefit of T's surviving spouse, S. On January 16, 1995, T's executor filed T's Form 706 on which the executor elects to treat the entire trust as qualified terminable...

  17. 26 CFR 1.405-2 - Deduction of contributions to qualified bond purchase plans.

    Science.gov (United States)

    2010-04-01

    ...), (9), and (10) and 404(e) shall also apply. (c) Accrual method taxpayers. In the case of a taxpayer using the accrual method of accounting, a contribution to a qualified bond purchase plan will be deemed paid on the last day of the year of accrual if— (1) During the taxable year of accrual the taxpayer...

  18. 12 CFR 24.6 - Examples of qualifying public welfare investments.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 1 2010-01-01 2010-01-01 false Examples of qualifying public welfare investments. 24.6 Section 24.6 Banks and Banking COMPTROLLER OF THE CURRENCY, DEPARTMENT OF THE TREASURY... finance small businesses or small farms, including minority- and women-owned small businesses or small...

  19. 48 CFR 809.270 - Qualified products for convenience/labor-saving foods.

    Science.gov (United States)

    2010-10-01

    ... convenience/labor-saving foods. 809.270 Section 809.270 Federal Acquisition Regulations System DEPARTMENT OF... 809.270 Qualified products for convenience/labor-saving foods. (a) Each VISN Nutrition and Food Service representative is authorized to establish a common VISN QPL for convenience and labor-saving foods...

  20. Return to Learning, Return to Work: Helping Low-Qualified Adults Out of Unemployment. Briefing Note

    Science.gov (United States)

    Cedefop - European Centre for the Development of Vocational Training, 2013

    2013-01-01

    Since 2008, the question uppermost in policymakers' minds has understandably been the explosive growth of youth unemployment. But the crisis has also severely affected low-qualified adults, who face the highest rate of unemployment across Europe. A recent Cedefop study confirms that work-based training programmes can address the particular needs…

  1. Indiana Teachers' Perspectives on Testing Accommodations for Limited English Proficient Students Taking the Graduation Qualifying Exam

    Science.gov (United States)

    Hetler, Angela Dawn

    2010-01-01

    This qualitative case study examines teachers' perspectives on testing accommodations for Limited English Proficient (LEP) students taking Indiana's Graduation Qualifying Exam (GQE). The Indiana Department of Education (IDOE) states that the purpose of testing accommodations is to "level the playing field" between LEP students and their…

  2. 78 FR 33233 - Patient Protection and Affordable Care Act; Establishment of Exchanges and Qualified Health Plans...

    Science.gov (United States)

    2013-06-04

    ... employee choice environment and to make the systems and operational changes required for SHOP enrollment... requested that HHS clarify how the transitional employee choice policy would affect the employer... qualified employees and their dependents and implements a transitional policy regarding employees' choice of...

  3. 38 CFR 21.6519 - Eligibility of qualified veterans for employment and counseling services.

    Science.gov (United States)

    2010-07-01

    ... veterans for employment and counseling services. 21.6519 Section 21.6519 Pensions, Bonuses, and Veterans... employment and counseling services. (a) General. A qualified veteran for whom vocational rehabilitation and achievenment of a vocational goal are reasonably feasible may be provided the employment and counseling...

  4. 6 CFR 25.6 - Procedures for designation of qualified anti-terrorism technologies.

    Science.gov (United States)

    2010-01-01

    ...-terrorism technologies. 25.6 Section 25.6 Domestic Security DEPARTMENT OF HOMELAND SECURITY, OFFICE OF THE SECRETARY REGULATIONS TO SUPPORT ANTI-TERRORISM BY FOSTERING EFFECTIVE TECHNOLOGIES § 25.6 Procedures for designation of qualified anti-terrorism technologies. (a) Application Procedure. Any person, firm or other...

  5. 26 CFR 1.1275-6 - Integration of qualifying debt instruments.

    Science.gov (United States)

    2010-04-01

    ... § 1.483-4 or § 1.1275-4(c) (certain contingent payment debt instruments issued for nonpublicly traded... the qualifying debt instrument. A financial instrument that hedges currency risk is not a § 1.1275-6... denominated in pounds, the swap hedges only interest rate risk, not currency risk. Therefore, the transaction...

  6. Preparing Special Educators Highly Qualified in Content: Alternative Route Certification for Unlicensed Teachers in Rural Georgia

    Science.gov (United States)

    Childre, Amy L.

    2014-01-01

    The shortage of highly qualified special educators is most pronounced in rural schools serving populations characterized by poverty, low achievement, disability, and cultural diversity. The result is often untrained teachers serving students with the greatest education needs. This article describes efforts by a university in rural middle Georgia…

  7. 26 CFR 1.25-4T - Qualified mortgage credit certificate program (Temporary).

    Science.gov (United States)

    2010-04-01

    ... designed to ensure that only those individuals who could not otherwise purchase a residence will benefit... may be issued in connection with the qualified rehabilitation of a residence part of the cost of which... surrendered to other issuers during the calendar year, (v) The date and amount of any previous elections under...

  8. 26 CFR 1.48-12 - Qualified rehabilitated building; expenditures incurred after December 31, 1981.

    Science.gov (United States)

    2010-04-01

    ..., DEPARTMENT OF THE TREASURY INCOME TAX INCOME TAXES Rules for Computing Credit for Investment in Certain Depreciable Property § 1.48-12 Qualified rehabilitated building; expenditures incurred after December 31, 1981... section 48(g) and this section) is section 38 property. Property that is section 38 property by reason of...

  9. 47 CFR Appendix A to Part 400 - Minimum Grant Awards Available to Qualifying States

    Science.gov (United States)

    2010-10-01

    ... ADMINISTRATION, DEPARTMENT OF COMMERCE, AND NATIONAL HIGHWAY TRAFFIC SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION E-911 GRANT PROGRAM Pt. 400, App. A Appendix A to Part 400—Minimum Grant Awards Available to Qualifying States State name Minimum E-911grant award Alabama $686,230.25 Alaska 500,000.00 American Samoa...

  10. An Exploration of Changes in Thinking in the Transition from Student Teacher to Newly Qualified Teacher

    Science.gov (United States)

    Haggarty, Linda; Postlethwaite, Keith

    2012-01-01

    For newly qualified teachers (NQTs), the induction period of support is an important phase which has the potential to deepen learning that has already taken place in initial teacher education (ITE) as well as preparing the NQT for future learning. A particularly crucial time in the induction process is the first term of teaching, when NQTs are…

  11. 40 CFR 35.4190 - How does my group identify a qualified technical advisor?

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 1 2010-07-01 2010-07-01 false How does my group identify a qualified technical advisor? 35.4190 Section 35.4190 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GRANTS...: (1) Demonstrated knowledge of hazardous or toxic waste issues, relocation issues, redevelopment...

  12. 26 CFR 1.856-9 - Treatment of certain qualified REIT subsidiaries.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 9 2010-04-01 2010-04-01 false Treatment of certain qualified REIT subsidiaries. 1.856-9 Section 1.856-9 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Real Estate Investment Trusts § 1.856-9 Treatment of certain...

  13. Tricks of the trade: time management tips for newly qualified doctors.

    Science.gov (United States)

    Offiah, Gozie; Doherty, Eva

    2018-03-01

    The transition from medical student to doctor is an important milestone. The discovery that their time is no longer their own and that the demands of their job are greater than the time they have available is extremely challenging. At a recent surgical boot camp training programme, 60 first-year surgical trainees who had just completed their internship were invited to reflect on the lessons learnt regarding effective time management and to recommend tips for their newly qualified colleagues. They were asked to identify clinical duties that were considered urgent and important using the time management matrix and the common time traps encountered by newly qualified doctors. The surgical trainees identified several practical tips that ranged from writing a priority list to working on relationships within the team. These tips are generic and so applicable to all newly qualified medial doctors. We hope that awareness of these tips from the outset as against learning them through experience will greatly assist newly qualified doctors. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  14. Pedagogical characteristics of the systems for assessing the technical and tactical skills of qualified volleyball players

    Directory of Open Access Journals (Sweden)

    Мykhailo Oliinyk

    2017-12-01

    Full Text Available Purpose: on the basis of the analysis of the special literature and their own experimental studies, to determine the characteristics of the systems for assessing the technical and tactical skills of qualified volleyball players. Material & Methods: study was conducted on the basis of the material of the national team of Zaporozhye State Medical University on volleyball. Analysis and generalization of the data of scientific and methodical literature are used; registration, analysis and interpretation of indicators of technical and tactical actions of qualified volleyball players in training and competitive processes; pedagogical observations; algorithms for calculating the quantitative and qualitative indicators of technical and tactical skill in volleyball; methods of mathematical statistics. Results: data on the existence of a strong statistical relationship between the systems of assessment of technical and tactical skills of qualified volleyball players, taking into account the specificity of the game role. Conclusion: shows the specifics of the application of various systems for assessing the technical and tactical skills of qualified volleyball players, taking into account the factors of the game role, noted the methodological features of special analysis and interpretation of indicators of technical and tactical actions in the adversarial process.

  15. 40 CFR 73.81 - Qualified conservation measures and renewable energy generation.

    Science.gov (United States)

    2010-07-01

    ...)) without increasing the use by the customer of any fuel other than qualified renewable energy, industrial waste heat, or, pursuant to paragraph (b)(5) of this section, industrial waste gases; (ii) Is... pursuant to § 73.82(c); or (5) Utilization of industrial waste gases, unless the applicant has certified...

  16. Encounters of Newly Qualified Teachers with Micro-Politics in Primary Schools in Zimbabwe

    Science.gov (United States)

    Magudu, Snodia; Gumbo, Mishack

    2017-01-01

    This article demonstrates, through the example of Zimbabwe, the complexities of micro-political learning during induction. It reports on the experiences of ten newly qualified teachers with micro-politics or power relations in their schools during induction and locates these experiences within the broader context of their professional development.…

  17. Skills upgrading for newly qualified surgeon: Is the district hospital in ...

    African Journals Online (AJOL)

    With adequate surgical activity in these settings, the trainee's confidence is boosted to the point of independence. This skill acquisition is a continuous process, especially so for those who qualify from an exit training program like the one at the University of Nairobi (UON). The continuity is maintained for two years after.

  18. 26 CFR 1.1202-2 - Qualified small business stock; effect of redemptions.

    Science.gov (United States)

    2010-04-01

    ... purchased from the decedent's estate, beneficiary (whether by bequest or lifetime gift), heir, surviving.... Stock acquired by a taxpayer is not qualified small business stock if, in one or more purchases during... corporation purchases (directly or indirectly) more than a de minimis amount of its stock from the taxpayer or...

  19. 26 CFR 301.6361-1 - Collection and administration of qualified taxes.

    Science.gov (United States)

    2010-04-01

    ... chapter 1 (and the civil and criminal sanctions provided by subtitle F, or by title 18 of the United States Code (relating to crimes and criminal procedure), with respect to such collection and... for tax of another State or political subdivision—(i) In general. A credit allowable under a qualified...

  20. 76 FR 71450 - Deduction for Qualified Film and Television Production Costs; Correction

    Science.gov (United States)

    2011-11-18

    ... Deduction for Qualified Film and Television Production Costs; Correction AGENCY: Internal Revenue Service... Register on Wednesday, October 19, 2011 (76 FR 64816) relating to deductions for the cost of producing film and television productions. DATES: This correction is effective on November 18, 2011, and is...

  1. 76 FR 64879 - Deduction for Qualified Film and Television Production Costs

    Science.gov (United States)

    2011-10-19

    ... Deduction for Qualified Film and Television Production Costs AGENCY: Internal Revenue Service (IRS... regulations relating to deductions for the costs of producing film and television productions. Those temporary... production costs. (a) * * * (1) * * * (ii) [The text of this proposed amendment to Sec. 1.181- 1(a)(1)(ii) is...

  2. 42 CFR 405.2462 - Payment for rural health clinic and Federally qualified health center services.

    Science.gov (United States)

    2010-10-01

    ... integral and subordinate part of a hospital, skilled nursing facility or home health agency participating... 42 Public Health 2 2010-10-01 2010-10-01 false Payment for rural health clinic and Federally qualified health center services. 405.2462 Section 405.2462 Public Health CENTERS FOR MEDICARE & MEDICAID...

  3. Newly Qualified Teachers' Professional Digital Competence: Implications for Teacher Education

    Science.gov (United States)

    Gudmundsdottir, Greta Björk; Hatlevik, Ove Edvard

    2018-01-01

    The professional digital competence (PDC) of teachers is of growing importance in classrooms, now that digital resources and digital media are becoming important parts of teachers' everyday practice. This study explores how newly qualified teachers are prepared to use information and communication technology (ICT) in their initial teacher…

  4. The Utilisation of Highly Qualified Personnel. Venice Conference, 25th-27th October 1971.

    Science.gov (United States)

    Organisation for Economic Cooperation and Development, Paris (France).

    This document reports the highlights of an international conference designed to study the utilization of highly qualified personnel. Following the opening addresses by various participants, discussion documents and basic reports are presented. Four areas of concern include the state of employment and employment policy, the conditions and aims of…

  5. 77 FR 462 - Regulation of Fuels and Fuel Additives: Identification of Additional Qualifying Renewable Fuel...

    Science.gov (United States)

    2012-01-05

    ... sources--qualifying as cellulosic biofuel: [cir] Thermochemical pyrolysis. [cir] Thermochemical... covercrops, algal oil, biogenic waste oils/fats/greases, non-food grade corn oil, Canola/rapeseed oil, and... from annual covercrops, algal oil, biogenic waste oils/fats/greases, or non- food grade corn oil using...

  6. 49 CFR 213.305 - Designation of qualified individuals; general qualifications.

    Science.gov (United States)

    2010-10-01

    ... college level engineering program, supplemented by special on the job training emphasizing the techniques... engineering program, supplemented by special on the job training emphasizing the techniques to be employed in... 49 Transportation 4 2010-10-01 2010-10-01 false Designation of qualified individuals; general...

  7. 47 CFR 13.201 - Qualifying for a commercial operator license or endorsement.

    Science.gov (United States)

    2010-10-01

    ... class of license or endorsement specified below must pass, or otherwise receive credit for, the... endorsement. 13.201 Section 13.201 Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL COMMERCIAL RADIO OPERATORS Examination System § 13.201 Qualifying for a commercial operator license or endorsement...

  8. 3 CFR - Medicare Demonstration To Test Medical Homes in Federally Qualified Health Centers

    Science.gov (United States)

    2010-01-01

    ... buy equipment, modernize clinic facilities, expand into new facilities, and adopt or expand the use of... interdisciplinary teams to treat the “whole patient” and focus on chronic disease management to reduce the use of costlier providers of care, such as emergency rooms and hospitals. Federally qualified health centers...

  9. 42 CFR 423.884 - Requirements for qualified retiree prescription drug plans.

    Science.gov (United States)

    2010-10-01

    ... actuary who is a member of the American Academy of Actuaries. Applicants may use qualified outside actuaries, including (but not limited to) actuaries employed by the plan administrator or an insurer providing benefits under the plan. If an applicant uses an outside actuary, the attestation can be submitted...

  10. 26 CFR 1.381(c)(25)-1 - Deficiency dividend of a qualified investment entity.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 4 2010-04-01 2010-04-01 false Deficiency dividend of a qualified investment entity. 1.381(c)(25)-1 Section 1.381(c)(25)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Insolvency Reorganizations § 1.381(c)(25...

  11. 3 CFR 13495 - Executive Order 13495 of January 30, 2009. Nondisplacement of Qualified Workers Under Service...

    Science.gov (United States)

    2010-01-01

    ... contractor and its subcontractors shall, except as otherwise provided herein, in good faith offer those... by law, regulations in the Federal Acquisition Regulation to provide for inclusion of the contract.... Nondisplacement of Qualified Workers Under Service Contracts 13495 Order 13495 Presidential Documents Executive...

  12. 76 FR 58460 - Information Collection; Qualified Products List for Long-Term Retardant for Wildland Firefighting

    Science.gov (United States)

    2011-09-21

    ... adequate types and quantities of qualified fire chemical products available to accomplish fire management... made from these lists. To initiate an evaluation, product manufacturers (or authorized suppliers) enter... Substances and Their Threshold Planning Quantities''. A risk assessment, performed at manufacturer expense...

  13. The Importance of School Leaders' Engagement in Socialising Newly Qualified Teachers into the Teaching Profession

    Science.gov (United States)

    Engvik, Gunnar; Emstad, Anne Berit

    2017-01-01

    This article focuses on the importance of school leaders' commitment to socialising newly qualified teachers (NQTs) into the teaching profession. Framed by a social constructivist perspective, the article is based on four challenges novice teachers face as described by four school leaders. The aim is to illuminate how school leaders have…

  14. 26 CFR 1.163-11T - Allocation of certain prepaid qualified mortgage insurance premiums (temporary).

    Science.gov (United States)

    2010-04-01

    ... insurance premiums (temporary). 1.163-11T Section 1.163-11T Internal Revenue INTERNAL REVENUE SERVICE... insurance premiums (temporary). (a) Allocation—(1) In general. As provided in section 163(h)(3)(E), premiums... section applies whether the qualified mortgage insurance premiums are paid in cash or are financed...

  15. Transient fluctuations of intracellular zinc ions in cell proliferation

    Energy Technology Data Exchange (ETDEWEB)

    Li, Yuan [Division of Human Nutrition, Department of Preventive Medicine and Community Health, The University of Texas Medical Branch, Galveston, TX 77555 (United States); Maret, Wolfgang, E-mail: womaret@utmb.edu [Division of Human Nutrition, Department of Preventive Medicine and Community Health, The University of Texas Medical Branch, Galveston, TX 77555 (United States); Department of Anesthesiology, The University of Texas Medical Branch, Galveston, TX 77555 (United States)

    2009-08-15

    Zinc is essential for cell proliferation, differentiation, and viability. When zinc becomes limited for cultured cells, DNA synthesis ceases and the cell cycle is arrested. The molecular mechanisms of actions of zinc are believed to involve changes in the availability of zinc(II) ions (Zn{sup 2+}). By employing a fluorescent Zn{sup 2+} probe, FluoZin-3 acetoxymethyl ester, intracellular Zn{sup 2+} concentrations were measured in undifferentiated and in nerve growth factor (NGF)-differentiated rat pheochromocytoma (PC12) cells. Intracellular Zn{sup 2+} concentrations are pico- to nanomolar in PC12 cells and are higher in the differentiated than in the undifferentiated cells. When following cellular Zn{sup 2+} concentrations for 48 h after the removal of serum, a condition that is known to cause cell cycle arrest, Zn{sup 2+} concentrations decrease after 30 min but, remarkably, increase after 1 h, and then decrease again to about one half of the initial concentration. Cell proliferation, measured by an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, decreases after both serum starvation and zinc chelation. Two peaks of Zn{sup 2+} concentrations occur within one cell cycle: one early in the G1 phase and the other in the late G1/S phase. Thus, fluctuations of intracellular Zn{sup 2+} concentrations and established modulation of phosphorylation signaling, via an inhibition of protein tyrosine phosphatases at commensurately low Zn{sup 2+} concentrations, suggest a role for Zn{sup 2+} in the control of the cell cycle. Interventions targeted at these picomolar Zn{sup 2+} fluctuations may be a way of controlling cell growth in hyperplasia, neoplasia, and diseases associated with aberrant differentiation.

  16. Polymeric gel nanoparticle pH sensors for intracellular measurements

    OpenAIRE

    Almdal, Kristoffer; Andresen, Thomas Lars; Benjaminsen, Rikke Vicki; Christensen, Nynne Meyn; Henriksen, Jonas Rosager; Sun, Honghao

    2011-01-01

    Precise measurements of pH in cells and intracellular compartments are of importance to both the fundamental understanding of metabolism and to the development of drugs that are released from the endosomes-lysome pathway. We have developed polymer gel nanoparticles as carriers of covalently bound fluorophores for ratiometric measurements of pH. One pH insensitive fluorophore serves as a reference while one or more pH sensitive fluorophores serve to give the desired pH dependence of the output...

  17. Intracellular transport driven by cytoskeletal motors: General mechanisms and defects

    Science.gov (United States)

    Appert-Rolland, C.; Ebbinghaus, M.; Santen, L.

    2015-09-01

    Cells are the elementary units of living organisms, which are able to carry out many vital functions. These functions rely on active processes on a microscopic scale. Therefore, they are strongly out-of-equilibrium systems, which are driven by continuous energy supply. The tasks that have to be performed in order to maintain the cell alive require transportation of various ingredients, some being small, others being large. Intracellular transport processes are able to induce concentration gradients and to carry objects to specific targets. These processes cannot be carried out only by diffusion, as cells may be crowded, and quite elongated on molecular scales. Therefore active transport has to be organized. The cytoskeleton, which is composed of three types of filaments (microtubules, actin and intermediate filaments), determines the shape of the cell, and plays a role in cell motion. It also serves as a road network for a special kind of vehicles, namely the cytoskeletal motors. These molecules can attach to a cytoskeletal filament, perform directed motion, possibly carrying along some cargo, and then detach. It is a central issue to understand how intracellular transport driven by molecular motors is regulated. The interest for this type of question was enhanced when it was discovered that intracellular transport breakdown is one of the signatures of some neuronal diseases like the Alzheimer. We give a survey of the current knowledge on microtubule based intracellular transport. Our review includes on the one hand an overview of biological facts, obtained from experiments, and on the other hand a presentation of some modeling attempts based on cellular automata. We present some background knowledge on the original and variants of the TASEP (Totally Asymmetric Simple Exclusion Process), before turning to more application oriented models. After addressing microtubule based transport in general, with a focus on in vitro experiments, and on cooperative effects in the

  18. Dependence of cerebral arterial contractions on intracellularly stored Ca++.

    Science.gov (United States)

    Sasaki, T; Kassell, N F; Zuccarello, M

    1986-01-01

    The purpose of the present study was to evaluate the dependence of the arterial contractions induced by different vasoactive agents upon intracellularly stored calcium in canine versus monkey cerebral arteries. The potency for inducing contractions in Ca++-free media was in the order of 9,11-epithio-11,12-metano-thromboxane A2 (STXA2) greater than prostaglandin F2 alpha (PGF2 alpha) much greater than serotonin greater than K+ in canine basilar arteries, and STXA2 greater than PGF2 alpha much greater than serotonin = K+ in monkey basilar arteries.

  19. Oxygen effect and intracellular oxygen content (adaptation hypothesis)

    Energy Technology Data Exchange (ETDEWEB)

    Yarmonenko, S P; Ehpshtejn, I M [Akademiya Meditsinskikh Nauk SSSR, Moscow. Onkologicheskij Tsentr

    1977-01-01

    Experimental data indicating that a radiomodifying action of hypoxia is dependent on the ''prehistory'' of the irradiated object are considered. This dependence manifests itself in a decreased protective action of acute hypoxia on the hypoxia-adapted objects. To explain this a hypothesis is proposed connecting a degree of cell radiosensitivity modification, determined by the oxygen effect, with the intracellular oxygen content. The latter, in accord with current ideas, is regulated by variations in the diffusion resistance to oxygen shown by the cytoplasmic membranes depending on the energy level of the cell and the degree of its oxygenation.

  20. Oxygen effect and intracellular oxygen content (adaptation hypothesis)

    International Nuclear Information System (INIS)

    Yarmonenko, S.P.; Ehpshtejn, I.M.

    1977-01-01

    Experimental data indicating that a radiomodifying action of hypoxia is dependent on the ''prehistory'' of the irradiated object are considered. This dependence manifests itself in a decreased protective action of acute hypoxia on the hypoxia-adapted objects. To explain this a hypothesis is proposed connecting a degree of cell radiosensitivity modification, determined by the oxygen effect, with the intracellular oxygen content. The latter, in accord with current ideas, is regulated by variations in the diffusion resistance to oxygen shown by the cytoplasmic membranes depending on the energy level of the cell and the degree of its oxygenation

  1. Subcellular site and nature of intracellular cadmium in plants

    International Nuclear Information System (INIS)

    Wagner, G.J.

    1979-01-01

    The mechanisms underlying heavy metal accumulation, toxicity, and tolerance in higher plants are poorly understood. Since subcellular processes are undoubtedly involved in all these phenomena, it is of interest to study the extent, subcellular site and nature of intracellularly accumulated cadmium in higher plants. Whole plants supplied 109 CdCl 2 or 112 CdSO 4 accumulated Cd into roots and aerial tissues. Preparation of protoplasts from aerial tissues followed by subcellular fractionation of the protoplasts to obtain intact vacuoles, chloroplasts and cytosol revealed the presence of Cd in the cytosol but not in vacuoles or chloroplasts. No evidence was obtained for the production of volatile Cd complexes in tobacco

  2. Job satisfaction of overseas-qualified nurses working in Australian hospitals.

    Science.gov (United States)

    Timilsina Bhandari, K K; Xiao, L D; Belan, I

    2015-03-01

    The aims of this study were to explore factors associated with the job satisfaction of overseas-qualified nurses working in public hospitals in South Australia and to compare whether factors associated with job satisfaction of overseas nurses from English-speaking backgrounds differed from those from non-English-speaking backgrounds. Overseas-qualified nurses have become an essential part of the nursing workforce in Australia. Although this nurse population has different expectations and values in relation to their jobs when compared with local nurses, studies on job satisfaction among overseas nurses are scarce. A cross-sectional survey using the Job Satisfaction of Overseas-Qualified Nurses questionnaire was conducted in five major public hospitals in South Australia. One hundred and fifty-one overseas-qualified nurses completed the questionnaire. Four factors were found to influence job satisfaction: Supportive work environment, interpersonal relationships, communication in English, and salary and salary-related benefits. Communication in English was the predominant factor that was associated with job satisfaction in nurses from non-English-speaking backgrounds. This group of nurses also showed a negative correlation between length of stay in Australia and satisfaction with their work environment. Participants' responses to open-ended questions revealed issues relating to discrimination and racism. Supportive work environment, interpersonal relationships, communication in English, and salary and salary-related benefits were major factors associated with job satisfaction in overseas-qualified nurses in this study. Nurses from non-English-speaking backgrounds faced additional challenges in communication in the workplace and in dealing with issues of discrimination and racism. Nurses from non-English-speaking backgrounds need to be supported early in their employment, especially with their communication skills. Consideration also needs to be given to the education

  3. 29 CFR 2520.104-46 - Waiver of examination and report of an independent qualified public accountant for employee...

    Science.gov (United States)

    2010-07-01

    ... qualified public accountant for employee benefit plans with fewer than 100 participants. 2520.104-46 Section... Requirements § 2520.104-46 Waiver of examination and report of an independent qualified public accountant for... public accountant to conduct an examination of the financial statements of the plan; (2) Include within...

  4. 12 CFR 617.7315 - What records must the qualified lender maintain on behalf of the CRC?

    Science.gov (United States)

    2010-01-01

    ... on behalf of the CRC? 617.7315 Section 617.7315 Banks and Banking FARM CREDIT ADMINISTRATION FARM... must the qualified lender maintain on behalf of the CRC? A qualified lender must maintain a complete file of all requests for CRC reviews, including participation in state mediation programs, the minutes...

  5. 26 CFR 20.2039-2 - Annuities under “qualified plans” and section 403(b) annuity contracts.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 14 2010-04-01 2010-04-01 false Annuities under âqualified plansâ and section 403(b) annuity contracts. 20.2039-2 Section 20.2039-2 Internal Revenue INTERNAL REVENUE SERVICE... AUGUST 16, 1954 Gross Estate § 20.2039-2 Annuities under “qualified plans” and section 403(b) annuity...

  6. Crystallographic study of FABP5 as an intracellular endocannabinoid transporter

    International Nuclear Information System (INIS)

    Sanson, Benoît; Wang, Tao; Sun, Jing; Wang, Liqun; Kaczocha, Martin; Ojima, Iwao; Deutsch, Dale; Li, Huilin

    2014-01-01

    FABP5 was recently found to intracellularly transport endocannabinoid signaling lipids. The structures of FABP5 complexed with two endocannabinoids and an inhibitor were solved. Human FABP5 was found to dimerize via a domain-swapping mechanism. This work will help in the development of inhibitors to raise endocannabinoid levels. In addition to binding intracellular fatty acids, fatty-acid-binding proteins (FABPs) have recently been reported to also transport the endocannabinoids anandamide (AEA) and 2-arachidonoylglycerol (2-AG), arachidonic acid derivatives that function as neurotransmitters and mediate a diverse set of physiological and psychological processes. To understand how the endocannabinoids bind to FABPs, the crystal structures of FABP5 in complex with AEA, 2-AG and the inhibitor BMS-309403 were determined. These ligands are shown to interact primarily with the substrate-binding pocket via hydrophobic interactions as well as a common hydrogen bond to the Tyr131 residue. This work advances our understanding of FABP5–endocannabinoid interactions and may be useful for future efforts in the development of small-molecule inhibitors to raise endocannabinoid levels

  7. Imaging the intracellular degradation of biodegradable polymer nanoparticles

    Directory of Open Access Journals (Sweden)

    Anne-Kathrin Barthel

    2014-10-01

    Full Text Available In recent years, the development of smart drug delivery systems based on biodegradable polymeric nanoparticles has become of great interest. Drug-loaded nanoparticles can be introduced into the cell interior via endocytotic processes followed by the slow release of the drug due to degradation of the nanoparticle. In this work, poly(L-lactic acid (PLLA was chosen as the biodegradable polymer. Although common degradation of PLLA has been studied in various biological environments, intracellular degradation processes have been examined only to a very limited extent. PLLA nanoparticles with an average diameter of approximately 120 nm were decorated with magnetite nanocrystals and introduced into mesenchymal stem cells (MSCs. The release of the magnetite particles from the surface of the PLLA nanoparticles during the intracellular residence was monitored by transmission electron microscopy (TEM over a period of 14 days. It was demonstrated by the release of the magnetite nanocrystals from the PLLA surface that the PLLA nanoparticles do in fact undergo degradation within the cell. Furthermore, even after 14 days of residence, the PLLA nanoparticles were found in the MSCs. Additionally, the ultrastructural TEM examinations yield insight into the long term intercellular fate of these nanoparticles. From the statistical analysis of ultrastructural details (e.g., number of detached magnetite crystals, and the number of nanoparticles in one endosome, we demonstrate the importance of TEM studies for such applications in addition to fluorescence studies (flow cytometry and confocal laser scanning microscopy.

  8. Fluorescent nanosensors for intracellular measurements: synthesis, characterisation, calibration and measurement

    Directory of Open Access Journals (Sweden)

    Arpan Shailesh Desai

    2014-01-01

    Full Text Available Measurement of intracellular acidification is important for understanding fundamental biological pathways as well as developing effective therapeutic strategies. Fluorescent pH nanosensors are an enabling technology for real-time monitoring of intracellular acidification. The physicochemical characteristics of nanosensors can be engineered to target specific cellular compartments and respond to external stimuli. Therefore nanosensors represent a versatile approach for probing biological pathways inside cells. The fundamental components of nanosensors comprise a pH-sensitive fluorophore (signal transducer and a pH-insensitive reference fluorophore (internal standard immobilised in an inert non-toxic matrix. The inert matrix prevents interference of cellular components with the sensing elements as well as minimizing potentially harmful effects of some fluorophores on cell function. Fluorescent nanosensors are synthesised using standard laboratory equipment and are detectable by non-invasive widely accessibly imaging techniques. The outcomes of studies employing this technology are dependent on reliable methodology for performing measurements. In particular special consideration must be given to conditions for sensor calibration, uptake conditions and parameters for image analysis. We describe procedures for: 1 synthesis and characterisation of polyacrylamide and silica based nanosensors 2 nanosensor calibration and 3 performing measurements using fluorescence microscopy.

  9. Modeling nanoparticle uptake and intracellular distribution using stochastic process algebras

    Energy Technology Data Exchange (ETDEWEB)

    Dobay, M. P. D., E-mail: maria.pamela.david@physik.uni-muenchen.de; Alberola, A. Piera; Mendoza, E. R.; Raedler, J. O., E-mail: joachim.raedler@physik.uni-muenchen.de [Ludwig-Maximilians University, Faculty of Physics, Center for NanoScience (Germany)

    2012-03-15

    Computational modeling is increasingly important to help understand the interaction and movement of nanoparticles (NPs) within living cells, and to come to terms with the wealth of data that microscopy imaging yields. A quantitative description of the spatio-temporal distribution of NPs inside cells; however, it is challenging due to the complexity of multiple compartments such as endosomes and nuclei, which themselves are dynamic and can undergo fusion and fission and exchange their content. Here, we show that stochastic pi calculus, a widely-used process algebra, is well suited for mapping surface and intracellular NP interactions and distributions. In stochastic pi calculus, each NP is represented as a process, which can adopt various states such as bound or aggregated, as well as be passed between processes representing location, as a function of predefined stochastic channels. We created a pi calculus model of gold NP uptake and intracellular movement and compared the evolution of surface-bound, cytosolic, endosomal, and nuclear NP densities with electron microscopy data. We demonstrate that the computational approach can be extended to include specific molecular binding and potential interaction with signaling cascades as characteristic for NP-cell interactions in a wide range of applications such as nanotoxicity, viral infection, and drug delivery.

  10. Modeling nanoparticle uptake and intracellular distribution using stochastic process algebras

    International Nuclear Information System (INIS)

    Dobay, M. P. D.; Alberola, A. Piera; Mendoza, E. R.; Rädler, J. O.

    2012-01-01

    Computational modeling is increasingly important to help understand the interaction and movement of nanoparticles (NPs) within living cells, and to come to terms with the wealth of data that microscopy imaging yields. A quantitative description of the spatio-temporal distribution of NPs inside cells; however, it is challenging due to the complexity of multiple compartments such as endosomes and nuclei, which themselves are dynamic and can undergo fusion and fission and exchange their content. Here, we show that stochastic pi calculus, a widely-used process algebra, is well suited for mapping surface and intracellular NP interactions and distributions. In stochastic pi calculus, each NP is represented as a process, which can adopt various states such as bound or aggregated, as well as be passed between processes representing location, as a function of predefined stochastic channels. We created a pi calculus model of gold NP uptake and intracellular movement and compared the evolution of surface-bound, cytosolic, endosomal, and nuclear NP densities with electron microscopy data. We demonstrate that the computational approach can be extended to include specific molecular binding and potential interaction with signaling cascades as characteristic for NP-cell interactions in a wide range of applications such as nanotoxicity, viral infection, and drug delivery.

  11. Modeling nanoparticle uptake and intracellular distribution using stochastic process algebras

    Science.gov (United States)

    Dobay, M. P. D.; Alberola, A. Piera; Mendoza, E. R.; Rädler, J. O.

    2012-03-01

    Computational modeling is increasingly important to help understand the interaction and movement of nanoparticles (NPs) within living cells, and to come to terms with the wealth of data that microscopy imaging yields. A quantitative description of the spatio-temporal distribution of NPs inside cells; however, it is challenging due to the complexity of multiple compartments such as endosomes and nuclei, which themselves are dynamic and can undergo fusion and fission and exchange their content. Here, we show that stochastic pi calculus, a widely-used process algebra, is well suited for mapping surface and intracellular NP interactions and distributions. In stochastic pi calculus, each NP is represented as a process, which can adopt various states such as bound or aggregated, as well as be passed between processes representing location, as a function of predefined stochastic channels. We created a pi calculus model of gold NP uptake and intracellular movement and compared the evolution of surface-bound, cytosolic, endosomal, and nuclear NP densities with electron microscopy data. We demonstrate that the computational approach can be extended to include specific molecular binding and potential interaction with signaling cascades as characteristic for NP-cell interactions in a wide range of applications such as nanotoxicity, viral infection, and drug delivery.

  12. Molecular characterization of a novel intracellular ADP-ribosyl cyclase.

    Directory of Open Access Journals (Sweden)

    Dev Churamani

    2007-08-01

    Full Text Available ADP-ribosyl cyclases are remarkable enzymes capable of catalyzing multiple reactions including the synthesis of the novel and potent intracellular calcium mobilizing messengers, cyclic ADP-ribose and NAADP. Not all ADP-ribosyl cyclases however have been characterized at the molecular level. Moreover, those that have are located predominately at the outer cell surface and thus away from their cytosolic substrates.Here we report the molecular cloning of a novel expanded family of ADP-ribosyl cyclases from the sea urchin, an extensively used model organism for the study of inositol trisphosphate-independent calcium mobilization. We provide evidence that one of the isoforms (SpARC1 is a soluble protein that is targeted exclusively to the endoplasmic reticulum lumen when heterologously expressed. Catalytic activity of the recombinant protein was readily demonstrable in crude cell homogenates, even under conditions where luminal continuity was maintained.Our data reveal a new intracellular location for ADP-ribosyl cyclases and suggest that production of calcium mobilizing messengers may be compartmentalized.

  13. Light irradiation helps magnetotactic bacteria eliminate intracellular reactive oxygen species.

    Science.gov (United States)

    Li, Kefeng; Wang, Pingping; Chen, Chuanfang; Chen, Changyou; Li, Lulu; Song, Tao

    2017-09-01

    Magnetotactic bacteria (MTB) demonstrate photoresponse. However, little is known about the biological significance of this behaviour. Magnetosomes exhibit peroxidase-like activity and can scavenge reactive oxygen species (ROS). Magnetosomes extracted from the Magnetospirillum magneticum strain AMB-1 show enhanced peroxidase-like activity under illumination. The present study investigated the effects of light irradiation on nonmagnetic (without magnetosomes) and magnetic (with magnetosomes) AMB-1 cells. Results showed that light irradiation did not affect the growth of nonmagnetic and magnetic cells but significantly increased magnetosome synthesis and reduced intracellular ROS level in magnetic cells. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to analyse the expression level of magnetosome formation-associated genes (mamA, mms6, mms13 and mmsF) and stress-related genes (recA, oxyR, SOD, amb0664 and amb2684). Results showed that light irradiation upregulated the expression of mms6, mms13 and mmsF. Furthermore, light irradiation upregulated the expression of stress-related genes in nonmagnetic cells but downregulated them in magnetic cells. Additionally, magnetic cells exhibited stronger phototactic behaviour than nonmagnetic ones. These results suggested that light irradiation could heighten the ability of MTB to eliminate intracellular ROS and help them adapt to lighted environments. This phenomenon may be related to the enhanced peroxidase-like activity of magnetosomes under light irradiation. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

  14. Cell fate reprogramming by control of intracellular network dynamics

    Science.gov (United States)

    Zanudo, Jorge G. T.; Albert, Reka

    Identifying control strategies for biological networks is paramount for practical applications that involve reprogramming a cell's fate, such as disease therapeutics and stem cell reprogramming. Although the topic of controlling the dynamics of a system has a long history in control theory, most of this work is not directly applicable to intracellular networks. Here we present a network control method that integrates the structural and functional information available for intracellular networks to predict control targets. Formulated in a logical dynamic scheme, our control method takes advantage of certain function-dependent network components and their relation to steady states in order to identify control targets, which are guaranteed to drive any initial state to the target state with 100% effectiveness and need to be applied only transiently for the system to reach and stay in the desired state. We illustrate our method's potential to find intervention targets for cancer treatment and cell differentiation by applying it to a leukemia signaling network and to the network controlling the differentiation of T cells. We find that the predicted control targets are effective in a broad dynamic framework. Moreover, several of the predicted interventions are supported by experiments. This work was supported by NSF Grant PHY 1205840.

  15. Transient light-induced intracellular oxidation revealed by redox biosensor

    Energy Technology Data Exchange (ETDEWEB)

    Kolossov, Vladimir L., E-mail: viadimer@illinois.edu [Institute for Genomic Biology, University of Illinois at Urbana-Champaign, 1206 W. Gregory Drive, Urbana, IL 61801 (United States); Beaudoin, Jessica N. [Institute for Genomic Biology, University of Illinois at Urbana-Champaign, 1206 W. Gregory Drive, Urbana, IL 61801 (United States); Department of Animal Sciences, University of Illinois at Urbana-Champaign, 1207 W. Gregory Drive, Urbana, IL 61801 (United States); Hanafin, William P. [Institute for Genomic Biology, University of Illinois at Urbana-Champaign, 1206 W. Gregory Drive, Urbana, IL 61801 (United States); DiLiberto, Stephen J. [Institute for Genomic Biology, University of Illinois at Urbana-Champaign, 1206 W. Gregory Drive, Urbana, IL 61801 (United States); Department of Animal Sciences, University of Illinois at Urbana-Champaign, 1207 W. Gregory Drive, Urbana, IL 61801 (United States); Kenis, Paul J.A. [Institute for Genomic Biology, University of Illinois at Urbana-Champaign, 1206 W. Gregory Drive, Urbana, IL 61801 (United States); Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, 600 S. Mathews Avenue, Urbana, IL 61801 (United States); Rex Gaskins, H. [Institute for Genomic Biology, University of Illinois at Urbana-Champaign, 1206 W. Gregory Drive, Urbana, IL 61801 (United States); Department of Animal Sciences, University of Illinois at Urbana-Champaign, 1207 W. Gregory Drive, Urbana, IL 61801 (United States); Department of Pathobiology, University of Illinois at Urbana-Champaign, 2001 S. Lincoln Avenue, Urbana, IL 61801 (United States); Division of Nutritional Sciences, University of Illinois at Urbana-Champaign, 905 S. Goodwin Avenue, Urbana, IL 61801 (United States)

    2013-10-04

    Highlights: •Time-resolved live cell imaging revealed light-induced oxidation. •Only the roGFP probe fused with glutaredoxin reveals photooxidation. •The transient oxidation is rapidly reduced by the cytosolic antioxidant system. •Intracellular photooxidation is media-dependent. •Oxidation is triggered exclusively by exposure to short wavelength excitation. -- Abstract: We have implemented a ratiometric, genetically encoded redox-sensitive green fluorescent protein fused to human glutaredoxin (Grx1-roGFP2) to monitor real time intracellular glutathione redox potentials of mammalian cells. This probe enabled detection of media-dependent oxidation of the cytosol triggered by short wavelength excitation. The transient nature of light-induced oxidation was revealed by time-lapse live cell imaging when time intervals of less than 30 s were implemented. In contrast, transient ROS generation was not observed with the parental roGFP2 probe without Grx1, which exhibits slower thiol-disulfide exchange. These data demonstrate that the enhanced sensitivity of the Grx1-roGFP2 fusion protein enables the detection of short-lived ROS in living cells. The superior sensitivity of Grx1-roGFP2, however, also enhances responsiveness to environmental cues introducing a greater likelihood of false positive results during image acquisition.

  16. Detection of ubiquitinated huntingtin species in intracellular aggregates

    Directory of Open Access Journals (Sweden)

    Katrin eJuenemann

    2015-01-01

    Full Text Available Protein conformation diseases, including polyglutamine diseases, result from the accumulation and aggregation of misfolded proteins. Huntington’s disease is one of nine diseases caused by an expanded polyglutamine repeat within the affected protein and is hallmarked by intracellular inclusion bodies composed of aggregated N-terminal huntingtin fragments and other sequestered proteins. Fluorescence microscopy and filter trap assay are conventional methods to study protein aggregates, but cannot be used to analyze the presence and levels of post-translational modifications of aggregated huntingtin such as ubiquitination. Ubiquitination of proteins can be a signal for degradation and intracellular localization, but also affects protein activity and protein-protein interactions. The function of ubiquitination relies on its mono- and polymeric isoforms attached to protein substrates. Studying the ubiquitination pattern of aggregated huntingtin fragments offers an important possibility to understand huntingtin degradation and aggregation processes within the cell. For the identification of aggregated huntingtin and its ubiquitinated species, solubilization of the cellular aggregates is mandatory. Here we describe methods to identify post-translational modifications such as ubiquitination of aggregated mutant huntingtin. This approach is specifically described for use with mammalian cell culture and is suitable to study other disease-related proteins prone to aggregate.

  17. Downregulation of transferrin receptor surface expression by intracellular antibody

    International Nuclear Information System (INIS)

    Peng Jilin; Wu Sha; Zhao Xiaoping; Wang Min; Li Wenhan; Shen Xin; Liu Jing; Lei Ping; Zhu Huifen; Shen Guanxin

    2007-01-01

    To deplete cellular iron uptake, and consequently inhibit the proliferation of tumor cells, we attempt to block surface expression of transferrin receptor (TfR) by intracellular antibody technology. We constructed two expression plasmids (scFv-HAK and scFv-HA) coding for intracellular single-chain antibody against TfR with or without endoplasmic reticulum (ER) retention signal, respectively. Then they were transfected tumor cells MCF-7 by liposome. Applying RT-PCR, Western blotting, immunofluorescence microscopy and immunoelectron microscope experiments, we insure that scFv-HAK intrabody was successfully expressed and retained in ER contrasted to the secreted expression of scFv-HA. Flow cytometric analysis confirmed that the TfR surface expression was markedly decreased approximately 83.4 ± 2.5% in scFv-HAK transfected cells, while there was not significantly decrease in scFv-HA transfected cells. Further cell growth and apoptosis characteristics were evaluated by cell cycle analysis, nuclei staining and MTT assay. Results indicated that expression of scFv-HAK can dramatically induce cell cycle G1 phase arrest and apoptosis of tumor cells, and consequently significantly suppress proliferation of tumor cells compared with other control groups. For First time this study demonstrates the potential usage of anti-TfR scFv-intrabody as a growth inhibitor of TfR overexpressing tumors

  18. Cyanobacteria perceive nitrogen status by sensing intracellular 2-oxoglutarate levels.

    Science.gov (United States)

    Muro-Pastor, M I; Reyes, J C; Florencio, F J

    2001-10-12

    The regulatory circuits that control nitrogen metabolism are relatively well known in several bacterial model groups. However, much less is understood about how the nitrogen status of the cell is perceived in vivo. In cyanobacteria, the transcription factor NtcA is required for regulation (activation or repression) of an extensive number of genes involved in nitrogen metabolism. In contrast, how NtcA activity is regulated is largely unknown. Assimilation of ammonium by most microorganisms occurs through the sequential action of two enzymes: glutamine synthetase (GS) and glutamate synthase. Interestingly, regulation of the expression of NtcA-dependent genes in the cyanobacterium Synechocystis sp. PCC 6803 is altered in mutants with modified levels of GS activity. Two types of mutants were analyzed: glnA null mutants that lack GS type I and gif mutants unable to inactivate GS in the presence of ammonium. Changes in the intracellular pools of 19 different amino acids and the keto acid 2-oxoglutarate were recorded in wild-type and mutant strains under different nitrogen conditions. Our data strongly indicate that the nitrogen status in cyanobacteria is perceived as changes in the intracellular 2-oxoglutarate pool.

  19. Optochemokine Tandem for Light-Control of Intracellular Ca2.

    Directory of Open Access Journals (Sweden)

    Katrin Feldbauer

    Full Text Available An optochemokine tandem was developed to control the release of calcium from endosomes into the cytosol by light and to analyze the internalization kinetics of G-protein coupled receptors (GPCRs by electrophysiology. A previously constructed rhodopsin tandem was re-engineered to combine the light-gated Ca2+-permeable cation channel Channelrhodopsin-2(L132C, CatCh, with the chemokine receptor CXCR4 in a functional tandem protein tCXCR4/CatCh. The GPCR was used as a shuttle protein to displace CatCh from the plasma membrane into intracellular areas. As shown by patch-clamp measurements and confocal laser scanning microscopy, heterologously expressed tCXCR4/CatCh was internalized via the endocytic SDF1/CXCR4 signaling pathway. The kinetics of internalization could be followed electrophysiologically via the amplitude of the CatCh signal. The light-induced release of Ca2+ by tandem endosomes into the cytosol via CatCh was visualized using the Ca2+-sensitive dyes rhod2 and rhod2-AM showing an increase of intracellular Ca2+ in response to light.

  20. Horizontal Transmission of Intracellular Insect Symbionts via Plants

    Directory of Open Access Journals (Sweden)

    Ewa Chrostek

    2017-11-01

    Full Text Available Experimental evidence is accumulating that endosymbionts of phytophagous insects may transmit horizontally via plants. Intracellular symbionts known for manipulating insect reproduction and altering fitness (Rickettsia, Cardinium, Wolbachia, and bacterial parasite of the leafhopper Euscelidius variegatus have been found to travel from infected insects into plants. Other insects, either of the same or different species can acquire the symbiont from the plant through feeding, and in some cases transfer it to their progeny. These reports prompt many questions regarding how intracellular insect symbionts are delivered to plants and how they affect them. Are symbionts passively transported along the insect-plant-insect path, or do they actively participate in the process? How widespread are these interactions? How does symbiont presence influence the plant? And what conditions are required for the new infection to establish in an insect? From an ecological, evolutionary, and applied perspective, this mode of horizontal transmission could have profound implications if occurring frequently enough or if new stable symbiont infections are established. Transmission of symbionts through plants likely represents an underappreciated means of infection, both in terms of symbiont epidemiology and the movement of symbionts to new host species.

  1. Transient light-induced intracellular oxidation revealed by redox biosensor

    International Nuclear Information System (INIS)

    Kolossov, Vladimir L.; Beaudoin, Jessica N.; Hanafin, William P.; DiLiberto, Stephen J.; Kenis, Paul J.A.; Rex Gaskins, H.

    2013-01-01

    Highlights: •Time-resolved live cell imaging revealed light-induced oxidation. •Only the roGFP probe fused with glutaredoxin reveals photooxidation. •The transient oxidation is rapidly reduced by the cytosolic antioxidant system. •Intracellular photooxidation is media-dependent. •Oxidation is triggered exclusively by exposure to short wavelength excitation. -- Abstract: We have implemented a ratiometric, genetically encoded redox-sensitive green fluorescent protein fused to human glutaredoxin (Grx1-roGFP2) to monitor real time intracellular glutathione redox potentials of mammalian cells. This probe enabled detection of media-dependent oxidation of the cytosol triggered by short wavelength excitation. The transient nature of light-induced oxidation was revealed by time-lapse live cell imaging when time intervals of less than 30 s were implemented. In contrast, transient ROS generation was not observed with the parental roGFP2 probe without Grx1, which exhibits slower thiol-disulfide exchange. These data demonstrate that the enhanced sensitivity of the Grx1-roGFP2 fusion protein enables the detection of short-lived ROS in living cells. The superior sensitivity of Grx1-roGFP2, however, also enhances responsiveness to environmental cues introducing a greater likelihood of false positive results during image acquisition

  2. Crystallographic study of FABP5 as an intracellular endocannabinoid transporter

    Energy Technology Data Exchange (ETDEWEB)

    Sanson, Benoît; Wang, Tao [Brookhaven National Laboratory, Upton, NY 11973-5000 (United States); Sun, Jing; Wang, Liqun; Kaczocha, Martin [Stony Brook University, Stony Brook, NY 11794-5213 (United States); Ojima, Iwao [Stony Brook University, Stony Brook, NY 1794-3400 (United States); Stony Brook University, Stony Brook, NY 11794-3400 (United States); Deutsch, Dale, E-mail: dale.deutsch@stonybrook.edu [Stony Brook University, Stony Brook, NY 11794-5213 (United States); Stony Brook University, Stony Brook, NY 11794-3400 (United States); Li, Huilin, E-mail: dale.deutsch@stonybrook.edu [Brookhaven National Laboratory, Upton, NY 11973-5000 (United States); Stony Brook University, Stony Brook, NY 11794-5213 (United States); Stony Brook University, Stony Brook, NY 11794-3400 (United States)

    2014-02-01

    FABP5 was recently found to intracellularly transport endocannabinoid signaling lipids. The structures of FABP5 complexed with two endocannabinoids and an inhibitor were solved. Human FABP5 was found to dimerize via a domain-swapping mechanism. This work will help in the development of inhibitors to raise endocannabinoid levels. In addition to binding intracellular fatty acids, fatty-acid-binding proteins (FABPs) have recently been reported to also transport the endocannabinoids anandamide (AEA) and 2-arachidonoylglycerol (2-AG), arachidonic acid derivatives that function as neurotransmitters and mediate a diverse set of physiological and psychological processes. To understand how the endocannabinoids bind to FABPs, the crystal structures of FABP5 in complex with AEA, 2-AG and the inhibitor BMS-309403 were determined. These ligands are shown to interact primarily with the substrate-binding pocket via hydrophobic interactions as well as a common hydrogen bond to the Tyr131 residue. This work advances our understanding of FABP5–endocannabinoid interactions and may be useful for future efforts in the development of small-molecule inhibitors to raise endocannabinoid levels.

  3. CIRRHOSIS INDUCES APOPTOSIS IN RENAL TISSUE THROUGH INTRACELLULAR OXIDATIVE STRESS

    Directory of Open Access Journals (Sweden)

    Keli Cristina Simões da SILVEIRA

    2015-03-01

    Full Text Available Background Renal failure is a frequent and serious complication in patients with decompensated cirrhosis. Objectives We aimed to evaluate the renal oxidative stress, cell damage and impaired cell function in animal model of cirrhosis. Methods Secondary biliary cirrhosis was induced in rats by ligation of the common bile duct. We measured TBARS, ROS and mitochondrial membrane potential in kidney as markers of oxidative stress, and activities of the antioxidant enzymes. Relative cell viability was determined by trypan blue dye-exclusion assay. Annexin V-PE was used with a vital dye, 7-AAD, to distinguish apoptotic from necrotic cells and comet assay was used for determined DNA integrity in single cells. Results In bile duct ligation animals there was significant increase in the kidney lipoperoxidation and an increase of the level of intracellular ROS. There was too an increase in the activity of all antioxidant enzymes evaluated in the kidney. The percentage viability was above 90% in the control group and in bile duct ligation was 64.66% and the dominant cell death type was apoptosis. DNA damage was observed in the bile duct ligation. There was a decreased in the mitochondrial membrane potential from 71.40% ± 6.35% to 34.48% ± 11.40% in bile duct ligation. Conclusions These results indicate that intracellular increase of ROS cause damage in the DNA and apoptosis getting worse the renal function in cirrhosis.

  4. A missing piece of the workforce puzzle. The experiences of internationally qualified nurses in New Zealand: a literature review.

    Science.gov (United States)

    Jenkins, Brittany Lauren; Huntington, Annette

    2015-01-01

    To analyse the literature regarding the context and experiences of internationally qualified registered nurses, particularly Filipino and Indian nurses, who have transitioned to New Zealand. Internationally qualified nurses are a significant proportion of the nursing workforce in many developed countries including New Zealand. This is increasingly important as populations age, escalating demand for nurses. Understanding the internationally qualified nurse experience is required as this could influence migration in a competitive labour market. Examination of peer-reviewed research, policy and discussion documents, and technical reports. A systematic literature search sought articles published between 2001 and 2014 using Google Scholar, CINAHL, and Medline. Articles were critically appraised for relevance, transferability, and methodological rigour. Fifty-one articles met inclusion criteria and demonstrate internationally qualified nurses face significant challenges transitioning into New Zealand. The internationally qualified nurse experience of transitioning into a new country is little researched and requires further investigation.

  5. Intracellular Trafficking Modulation by Ginsenoside Rg3 Inhibits Brucella abortus Uptake and Intracellular Survival within RAW 264.7 Cells.

    Science.gov (United States)

    Huy, Tran Xuan Ngoc; Reyes, Alisha Wehdnesday Bernardo; Hop, Huynh Tan; Arayan, Lauren Togonon; Min, WonGi; Lee, Hu Jang; Rhee, Man Hee; Chang, Hong Hee; Kim, Suk

    2017-03-28

    Ginsenoside Rg3, a saponin extracted from ginseng, has various pharmacological and biological activities; however, its effects against Brucella infection are still unclear. Herein, the inhibitory effects of ginsenoside Rg3 against intracellular parasitic Brucella infection were evaluated through bacterial infection, adherence assays, and LAMP-1 colocalization, as well as immunoblotting and FACS for detecting MAPK signaling proteins and F-actin polymerization, respectively. The internalization, intracellular growth, and adherence of Brucella abortus in Rg3-treated RAW 264.7 cells were significantly decreased compared with the Rg3-untreated control. Furthermore, an apparent reduction of F-actin content and intensity of F-actin fluorescence in Rg3-treated cells was observed compared with B. abortus -infected cells without treatment by flow cytometry analysis and confocal microscopy, respectively. In addition, treating cells with Rg3 decreased the phosphorylation of MAPK signaling proteins such as ERK 1/2 and p38 compared with untreated cells. Moreover, the colocalization of B. abortus -containing phagosomes with LAMP-1 was markedly increased in Rg3-treated cells. These findings suggest that ginsenoside Rg3 inhibits B. abortus infection in mammalian cells and can be used as an alternative approach in the treatment of brucellosis.

  6. Mechanism of H. pylori intracellular entry: an in vitro study

    Directory of Open Access Journals (Sweden)

    Hui eLiu

    2012-03-01

    Full Text Available The majority of H. pylori reside on gastric epithelial cell surfaces and in the overlying mucus, but a small fraction of H. pylori enter host epithelial and immune cells. To explore the role of the nudA invasin in host cell entry, a ΔnudA deletion derivative of strain J99 was constructed and transformants were verified by PCR and by fluorescence in situ hybridization. AGS cells were inoculated with either wild type (WT strain J99 or its ΔnudA mutant to determine the fraction of bacteria that were bound to the cells and inside these cells using the gentamicin protection assay. We observed no significant difference between either the density of H. pylori bound to AGS cell membranes or the density of intracellular H. pylori. To further explore this finding, separate chambers of each culture were fixed in glutaraldehyde for transmission electron microscopy (TEM and immunogold TEM. This addition to the classical gentamicin assay demonstrated that there were significantly more intracellular, and fewer membrane-bound, H. pylori in WT-infected AGS cells than in ΔnudA allele infected cells. Thus, the sum of intracellular and membrane-bound H. pylori was similar in the two groups. Since no other similar TEM study has been performed, it is at present unknown whether our observations can be reproduced by others Taken together however, our observations suggest that the classical gentamicin protection assay is not sufficiently sensitive to analyze H. pylori cell entry and that the addition of TEM to the test demonstrate that nudA plays a role in H. pylori entry into AGS cells in vitro. In addition, deletion of the invasin gene appears to limit H. pylori to the AGS cell surface, where it may be partly protected against gentamicin. In contrast, this specific environment may render H. pylori more vulnerable to host defense and therapeutic intervention, and less prone to trigger normal immune, carcinogenic, and other developmental response pathways.

  7. Intracellular Crosslinking of Filoviral Nucleoproteins with Xintrabodies Restricts Viral Packaging

    Directory of Open Access Journals (Sweden)

    Tamarand Lee Darling

    2017-09-01

    Full Text Available Viruses assemble large macromolecular repeat structures that become part of the infectious particles or virions. Ribonucleocapsids (RNCs of negative strand RNA viruses are a prime example where repetition of nucleoprotein (NP along the genome creates a core polymeric helical scaffold that accommodates other nucleocapsid proteins including viral polymerase. The RNCs are transported through the cytosol for packaging into virions through association with viral matrix proteins at cell membranes. We hypothesized that RNC would be ideal targets for crosslinkers engineered to promote aberrant protein–protein interactions, thereby blocking their orderly transport and packaging. Previously, we had generated single-domain antibodies (sdAbs against Filoviruses that have all targeted highly conserved C-terminal regions of NP known to be repetitively exposed along the length of the RNCs of Marburgvirus (MARV and Ebolavirus (EBOV. Our crosslinker design consisted of dimeric sdAb expressed intracellularly, which we call Xintrabodies (X- for crosslinking. Electron microscopy of purified NP polymers incubated with purified sdAb constructs showed NP aggregation occurred in a genus-specific manner with dimeric and not monomeric sdAb. A virus-like particle (VLP assay was used for initial evaluation where we found that dimeric sdAb inhibited NP incorporation into VP40-based VLPs whereas monomeric sdAb did not. Inhibition of NP packaging was genus specific. Confocal microscopy revealed dimeric sdAb was diffuse when expressed alone but focused on pools of NP when the two were coexpressed, while monomeric sdAb showed ambivalent partition. Infection of stable Vero cell lines expressing dimeric sdAb specific for either MARV or EBOV NP resulted in smaller plaques and reduced progeny of cognate virus relative to wild-type Vero cells. Though the impact was marginal at later time-points, the collective data suggest that viral replication can be reduced by crosslinking

  8. LDL Receptors as Gateways for Intracellular Porphyrin Uptake

    International Nuclear Information System (INIS)

    Novick, S.; Laster, B.; Quastel, M.

    2004-01-01

    Boronated compounds are currently being studied for possible use in Boron Neutron Capture Therapy (BNCT). We found that one of these agents, BOPP (tetrakis-carborane-carboxylate, esters of 2,4-bis (a,b- dihydroxyethyl) deuteroporphyrin IX), could also be labeled with indium (In-BOPP) and, therefore, could also be used potentially to transport high Z atoms into tumor cell DNA for AET (Auger Electron Therapy). In order to assess the uptake of these agents into cells, the role of the LDL receptor in the intracellular accumulation of BOPP and In-BOPP was investigated. Pre-incubation of V-79 Chinese hamster cells in medium containing delipidized fetal bovine serum (FBS) markedly increased the subsequent uptake of intracellular boron transported by both BOPP and In-BOPP when compared with cells that had been pre-incubated with medium containing 10% normal FBS (lipidized). The increased uptake was characterized by elevated levels of receptor, and greater affinity was shown for both BOPP and In-BOPP, although less marked with the latter. Positive cooperativity was demonstrated by sigmoid saturation curves, Scatchard analysis and Hill plots. Increasing the amount of LDL in the incubation medium had a relatively small effect on the total accumulation of either indium or boron atoms inside the cell. Furthermore, chemical acetylation of LDL did not decrease the intracellular uptake of either boron or indium transported by BOPP or In-BOPP. It is thus concluded that BOPP and In-BOPP preferentially enter the cells directly by way of the LDL receptor and that only a small fraction of these molecules are transported into the cells indirectly using serum LDLs as their carriers. These data suggest a novel way of bringing greater amounts of boron and indium (and perhaps other agents) into tissues. Porphyrins can be used to transport different agents into tumor cells because they are tumor affinic molecules. Tumors express a higher number of LDL receptors than do most normal tissues

  9. Examining educators of the deaf as "highly qualified" teachers: roles and responsibilities under IDEA and NCLB.

    Science.gov (United States)

    Luft, Pamela

    2008-01-01

    Educators of the deaf were long considered "highly qualified" if they obtained state licensure from approved deaf education programs. But the No Child Left Behind Act (2001) redefined qualifications based on core academic content areas, without recognizing disability-specific expertise. NCLB's reauthorization will provide opportunities for examining definitions of "highly qualified" and ensuring that both general and special educators are appropriately prepared. Under the Individuals With Disabilities Education Act, educators of the deaf are primarily responsible for supporting implementation of each assigned student's individualized education program. When done skillfully and knowledgeably, IEP execution maximizes learning outcomes, and therefore would support NCLB mandates for improved student achievement. Instead of academic attainment alone, the primary "qualification" of educators of the deaf should be training and expertise in providing communication, learning, and assistive technology supports that allow access to academic content and, ultimately, address deaf students' historical underachievement.

  10. Improvement of technique of a jerk of the qualified sportswomen in weightlifting

    Directory of Open Access Journals (Sweden)

    Oksana Solodka

    2016-12-01

    Full Text Available Purpose: the improvement of technical preparedness of the qualified female weight-lifters taking into account the group model characteristics of technique of a jerk. Material & Methods: analysis and synthesis of data of scientific and methodical literature, pedagogical experiment, method of mathematical statistics. Results: the revealed shortcomings of technique of the execution of a jerk of the qualified female weight-lifters were, considering the group model characteristics of technique of a jerk of the first group of weight categories. Complexes of compensatory exercises were offered for the purpose of correction. Conclusions: the performance of complexes of the compensatory exercises, which are directed to the improvement of competitive exercise of a jerk of bar, allowed to increase the number of successfully executed raising of bar, for 10,3–19,8%, according to initial indicators of number of rises, and also to receive positive shifts in motive structure of a jerk of bar.

  11. Poking cells for efficient vector-free intracellular delivery

    Science.gov (United States)

    Wang, Ying; Yang, Yang; Yan, Li; Kwok, So Ying; Li, Wei; Wang, Zhigang; Zhu, Xiaoyue; Zhu, Guangyu; Zhang, Wenjun; Chen, Xianfeng; Shi, Peng

    2014-07-01

    Techniques for introducing foreign molecules and materials into living cells are of great value in cell biology research. A major barrier for intracellular delivery is to cross the cell membrane. Here we demonstrate a novel platform utilizing diamond nanoneedle arrays to facilitate efficient vector-free cytosolic delivery. Using our technique, cellular membrane is deformed by an array of nanoneedles with a force on the order of a few nanonewtons. We show that this technique is applicable to deliver a broad range of molecules and materials into different types of cells, including primary neurons in adherent culture. Especially, for delivering plasmid DNAs into neurons, our technique produces at least eightfold improvement (~45% versus ~1-5%) in transfection efficiency with a dramatically shorter experimental protocol, when compared with the commonly used lipofection approach. It is anticipated that our technique will greatly benefit basic research in cell biology and also a wide variety of clinical applications.

  12. Subcellular site and nature of intracellular cadmium in plants

    International Nuclear Information System (INIS)

    Wagner, G.J.

    1979-01-01

    The mechanisms underlying heavy metal accumulation, toxicity and tolerance in higher plants are poorly understood. Since subcellular processes are undoubtedly involved in all these phenomena, it is of interest to study the extent of, subcellular site of and nature of intracellularly accumulated cadmium in higher plants. Whole plants supplied 109 CdCl 2 or 112 CdSO 4 accumulated Cd into roots and aerial tissues. Preparation of protoplasts from aerial tissue followed by subcellular fractionation of the protoplasts to obtain intact vacuoles, chloroplasts and cytosol revealed the presence of Cd in the cytosol but not in vacuoles or chloroplasts. Particulate materials containing other cell components were also labeled. Of the 109 Cd supplied to plants, 2 to 10% was recovered in both cytosol preparations and in particulate materials. Cytosol contained proteinaceous--Cd complexes, free metal and low molecular weight Cd complexes. Labeling of protoplasts gave similar results. No evidence was obtained for the production of volatile Cd complexes in tobacco

  13. The intracellular cholesterol landscape: dynamic integrator of the immune response

    Science.gov (United States)

    Fessler, Michael B.

    2016-01-01

    Cholesterol has typically been considered an exogenous, disease-related factor in immunity; however, recent literature suggests that a paradigm shift is in order. Sterols are now recognized to ligate several immune receptors. Altered flux through the mevalonic acid synthesis pathway also appears to be a required event in the antiviral interferon response of macrophages and in the activation, proliferation, and differentiation of T cells. In this review, evidence is discussed that suggests an intrinsic, ‘professional’ role for sterols and oxysterols in macrophage and T cell immunity. Host defense may have been the original selection pressure behind the development of mechanisms for intracellular cholesterol homeostasis. Functional coupling between sterol metabolism and immunity has fundamental implications for health and disease. PMID:27692616

  14. Extraction of intracellular protein from Glaciozyma antarctica for proteomics analysis

    Science.gov (United States)

    Faizura, S. Nor; Farahayu, K.; Faizal, A. B. Mohd; Asmahani, A. A. S.; Amir, R.; Nazalan, N.; Diba, A. B. Farah; Muhammad, M. Nor; Munir, A. M. Abdul

    2013-11-01

    Two preparation methods of crude extracts of psychrophilic yeast Glaciozyma antarctica were compared in order to obtain a good recovery of intracellular proteins. Extraction with mechanical procedures using sonication was found to be more effective for obtaining good yield compare to alkaline treatment method. The procedure is simple, rapid, and produce better yield. A total of 52 proteins were identified by combining both extraction methods. Most of the proteins identified in this study involves in the metabolic process including glycolysis pathway, pentose phosphate pathway, pyruyate decarboxylation and also urea cyle. Several chaperons were identified including probable cpr1-cyclophilin (peptidylprolyl isomerase), macrolide-binding protein fkbp12 and heat shock proteins which were postulate to accelerate proper protein folding. Characteristic of the fundamental cellular processes inferred from the expressed-proteome highlight the evolutionary and functional complexity existing in this domain of life.

  15. Intracellular pH in rat pancreatic ducts

    DEFF Research Database (Denmark)

    Novak, I; Hug, M; Greger, R

    1997-01-01

    In order to study the mechanism of H+ and HCO3- transport in a HCO3- secreting epithelium, pancreatic ducts, we have measured the intracellular pH (pHi) in this tissue using the pH sensitive probe BCECF. We found that exposures of ducts to solutions containing acetate/acetic acid or NH4+/NH3...... buffers (20 mmol/l) led to pHi changes in accordance with entry of lipid-soluble forms of the buffers, followed by back-regulation of pHi by duct cells. In another type of experiment, changes in extracellular pH of solutions containing HEPES or HCO3-/CO2 buffers led to significant changes in pHi that did....... Under some conditions, these exchangers can be invoked to regulate cell pH....

  16. Intracellular Signaling Mediators in the Circulatory and Ventilatory Systems

    CERN Document Server

    Thiriet, Marc

    2013-01-01

    The volumes in this authoritative series present a multidisciplinary approach to modeling and simulation of flows in the cardiovascular and ventilatory systems, especially multiscale modeling and coupled simulations. The cardiovascular and respiratory systems are tightly coupled, as their primary function is to supply oxygen to and remove carbon dioxide from the body's cells. Because physiological conduits have deformable and reactive walls, macroscopic flow behavior and prediction must be coupled to phenomenological models of nano- and microscopic events in a corrector scheme of regulated mechanisms when the vessel lumen caliber varies markedly. Therefore, investigation of flows of blood and air in physiological conduits requires an understanding of the biology, chemistry, and physics of these systems together with the mathematical tools to describe their functioning. Volume 4 is devoted to major sets of intracellular mediators that transmit signals upon stimulation of cell-surface receptors.  Activation of...

  17. Intracellular mediators of potassium-induced aldosterone secretion

    International Nuclear Information System (INIS)

    Ganguly, A.; Chiou, S.; Davis, J.S.

    1990-01-01

    We have investigated the intracellular messengers of potassium in eliciting aldosterone secretion in calf adrenal glomerulosa cells since there were unresolved issues relating to the role of phosphoinositides, cAMP and protein kinases. We observed no evidence of hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP 2 ) in 3 H-inositol labeled alf adrenal cells or increase of cAMP in response to potassium. Addition of calcium channel blocker, nitrendipine after stimulating adrenal glomerulosa cells with potassium, markedly inhibited aldosterone secretion. A calmodulin inhibitor (W-7) produced greater reduction of aldosterone secretion than an inhibitor of protein kinase C (H-7). These results suggest that a rise in cytosolic free calcium concentration through voltage-dependent calcium channel and calmodulin are the critical determinants of aldosterone secretion stimulated by potassium

  18. ATPase and GTPase Tangos Drive Intracellular Protein Transport.

    Science.gov (United States)

    Shan, Shu-Ou

    2016-12-01

    The GTPase superfamily of proteins provides molecular switches to regulate numerous cellular processes. The 'GTPase switch' paradigm, in which external regulatory factors control the switch of a GTPase between 'on' and 'off' states, has been used to interpret the regulatory mechanism of many GTPases. However, recent work unveiled a class of nucleotide hydrolases that do not adhere to this classical paradigm. Instead, they use nucleotide-dependent dimerization cycles to regulate key cellular processes. In this review article, recent studies of dimeric GTPases and ATPases involved in intracellular protein targeting are summarized. It is suggested that these proteins can use the conformational plasticity at their dimer interface to generate multiple points of regulation, thereby providing the driving force and spatiotemporal coordination of complex cellular pathways. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Evaluating Nanoparticle Sensor Design for Intracellular pH Measurements

    DEFF Research Database (Denmark)

    Benjaminsen, Rikke Vicki; Sun, Honghao; Henriksen, Jonas Rosager

    2011-01-01

    Particle-based nanosensors have over the last decade been designed for optical fluorescent-based ratiometric measurements of pH in living cells. However, quantitative and time-resolved intracellular measurements of pH in endosomes and lysosomes using particle nanosensors is challenging...... and there is a need to improve measurement methodology. In the present paper, we have successfully carried out time resolved pH measurements in endosomes and lyosomes in living cells using nanoparticle sensors and show the importance of sensor choice for successful quantification. We have studied two nanoparticle...... quantification of pH is an unfortunate result when measuring pH too close to the limit of the sensitive range of the sensors. Triple-labeled nanosensors with a pH measurement range of 3.2-7.0, which was synthesized by adding two pH-sensitive fluorophores with different pKa to each sensor, seem to be a solution...

  20. Intracellular Events and Cell Fate in Filovirus Infection

    Directory of Open Access Journals (Sweden)

    Elena Ryabchikova

    2011-08-01

    Full Text Available Marburg and Ebola viruses cause a severe hemorrhagic disease in humans with high fatality rates. Early target cells of filoviruses are monocytes, macrophages, and dendritic cells. The infection spreads to the liver, spleen and later other organs by blood and lymph flow. A hallmark of filovirus infection is the depletion of non-infected lymphocytes; however, the molecular mechanisms leading to the observed bystander lymphocyte apoptosis are poorly understood. Also, there is limited knowledge about the fate of infected cells in filovirus disease. In this review we will explore what is known about the intracellular events leading to virus amplification and cell damage in filovirus infection. Furthermore, we will discuss how cellular dysfunction and cell death may correlate with disease pathogenesis.

  1. Intracellular Transport and Kinesin Superfamily Proteins: Structure, Function and Dynamics

    Science.gov (United States)

    Hirokawa, N.; Takemura, R.

    Using various molecular cell biological and molecular genetic approaches, we identified kinesin superfamily proteins (KIFs) and characterized their significant functions in intracellular transport, which is fundamental for cellular morphogenesis, functioning, and survival. We showed that KIFs not only transport various membranous organelles, proteins complexes and mRNAs fundamental for cellular functions but also play significant roles in higher brain functions such as memory and learning, determination of important developmental processes such as left-right asymmetry formation and brain wiring. We also elucidated that KIFs recognize and bind to their specific cargoes using scaffolding or adaptor protein complexes. Concerning the mechanism of motility, we discovered the simplest unique monomeric motor KIF1A and determined by molecular biophysics, cryoelectron microscopy and X-ray crystallography that KIF1A can move on a microtubule processively as a monomer by biased Brownian motion and by hydolyzing ATP.

  2. Purification and characterization of an intracellular peroxidase from Streptomyces cyaneus.

    OpenAIRE

    Mliki, A; Zimmermann, W

    1992-01-01

    An intracellular peroxidase (EC 1.11.1.7) from Streptomyces cyaneus was purified to homogeneity. The enzyme had a molecular weight of 185,000 and was composed of two subunits of equal size. It had an isoelectric point of 6.1. The enzyme had a peroxidase activity toward o-dianisidine with a Km of 17.8 microM and a pH optimum of 5.0. It also showed catalase activity with a Km of 2.07 mM H2O2 and a pH optimum of 8.0. The purified enzyme did not catalyze C alpha-C beta bond cleavage of 1,3-dihydr...

  3. Measuring intracellular redox conditions using GFP-based sensors

    DEFF Research Database (Denmark)

    Björnberg, Olof; Ostergaard, Henrik; Winther, Jakob R

    2006-01-01

    Recent years have seen the development of methods for analyzing the redox conditions in specific compartments in living cells. These methods are based on genetically encoded sensors comprising variants of Green Fluorescent Protein in which vicinal cysteine residues have been introduced at solvent......-exposed positions. Several mutant forms have been identified in which formation of a disulfide bond between these cysteine residues results in changes of their fluorescence properties. The redox sensors have been characterized biochemically and found to behave differently, both spectroscopically and in terms...... of redox properties. As genetically encoded sensors they can be expressed in living cells and used for analysis of intracellular redox conditions; however, which parameters are measured depends on how the sensors interact with various cellular redox components. Results of both biochemical and cell...

  4. 2011 Rita Schaffer lecture: nanoparticles for intracellular nucleic acid delivery.

    Science.gov (United States)

    Green, Jordan J

    2012-07-01

    Nanoparticles are a promising technology for delivery of new types of therapeutics. A polymer library approach has allowed engineering of polymeric particles that are particularly effective for the delivery of DNA and siRNA to human cells. Certain chemical structural motifs, degradable linkages, hydrophobicity, and biophysical properties are key for successful intracellular delivery. Small differences to biomaterial structure, and especially the type of degradable linkage in the polymers, can be critical for successful delivery of siRNA vs. DNA. Furthermore, subtle changes to biomaterial structure can facilitate cell-type gene delivery specificity between human brain cancer cells and healthy cells as well as between human retinal endothelial cells and epithelial cells. These polymeric nanoparticles are effective for nucleic acid delivery in a broad range of human cell types and have applications to regenerative medicine, ophthalmology, and cancer among many other biomedical research areas.

  5. Control of intracellular heme levels: Heme transporters and Heme oxygenases

    Science.gov (United States)

    Khan, Anwar A.; Quigley, John G.

    2011-01-01

    Heme serves as a co-factor in proteins involved in fundamental biological processes including oxidative metabolism, oxygen storage and transport, signal transduction and drug metabolism. In addition, heme is important for systemic iron homeostasis in mammals. Heme has important regulatory roles in cell biology, yet excessive levels of intracellular heme are toxic; thus, mechanisms have evolved to control the acquisition, synthesis, catabolism and expulsion of cellular heme. Recently, a number of transporters of heme and heme synthesis intermediates have been described. Here we review aspects of heme metabolism and discuss our current understanding of heme transporters, with emphasis on the function of the cell-surface heme exporter, FLVCR. Knockdown of Flvcr in mice leads to both defective erythropoiesis and disturbed systemic iron homeostasis, underscoring the critical role of heme transporters in mammalian physiology. PMID:21238504

  6. An active matter analysis of intracellular Active Transport

    Science.gov (United States)

    Wang, Bo; Chen, Kejia; Bae, Sung Chul; Granick, Steve

    2012-02-01

    Tens of thousands of fluorescence-based trajectories at nm resolution have been analyzed, regarding active transport along microtubules in living cells. The following picture emerges. Directed motion to pre-determined locations is certainly an attractive idea, but cannot be pre-programmed as to do so would sacrifice adaptability. The polarity of microtubules is inadequate to identify these directions in cells, and no other mechanism is currently known. We conclude that molecular motors carry cargo through disordered intracellular microtubule networks in a statistical way, with loud cellular ``noise'' both in directionality and speed. Programmed random walks describe how local 1D active transport traverses crowded cellular space efficiently, rapidly, minimizing the energy waste that would result from redundant activity. The mechanism of statistical regulation is not yet understood, however.

  7. Variety in intracellular diffusion during the cell cycle

    DEFF Research Database (Denmark)

    Selhuber-Unkel, C.; Yde, P.; Berg-Sørensen, Kirstine

    2009-01-01

    During the cell cycle, the organization of the cytoskeletal network undergoes dramatic changes. In order to reveal possible changes of the viscoelastic properties in the intracellular space during the cell cycle we investigated the diffusion of endogenous lipid granules within the fission yeast...... Schizosaccharomyces Pombe using optical tweezers. The cell cycle was divided into interphase and mitotic cell division, and the mitotic cell division was further subdivided in its stages. During all stages of the cell cycle, the granules predominantly underwent subdiffusive motion, characterized by an exponent...... a that is also linked to the viscoelastic moduli of the cytoplasm. The exponent a was significantly smaller during interphase than during any stage of the mitotic cell division, signifying that the cytoplasm was more elastic during interphase than during division. We found no significant differences...

  8. Liposome-based Formulation for Intracellular Delivery of Functional Proteins

    Directory of Open Access Journals (Sweden)

    Benoît Chatin

    2015-01-01

    Full Text Available The intracellular delivery of biologically active protein represents an important emerging strategy for both fundamental and therapeutic applications. Here, we optimized in vitro delivery of two functional proteins, the β-galactosidase (β-gal enzyme and the anti-cytokeratin8 (K8 antibody, using liposome-based formulation. The guanidinium-cholesterol cationic lipid bis (guanidinium-tren-cholesterol (BGTC (bis (guanidinium-tren-cholesterol combined to the colipid dioleoyl phosphatidylethanolamine (DOPE (dioleoyl phosphatidylethanolamine was shown to efficiently deliver the β-gal intracellularly without compromising its activity. The lipid/protein molar ratio, protein amount, and culture medium were demonstrated to be key parameters affecting delivery efficiency. The protein itself is an essential factor requiring selection of the appropriate cationic lipid as illustrated by low K8 binding activity of the anti-K8 antibody using guanidinium-based liposome. Optimization of various lipids led to the identification of the aminoglycoside lipid dioleyl succinyl paromomycin (DOSP associated with the imidazole-based helper lipid MM27 as a potent delivery system for K8 antibody, achieving delivery in 67% of HeLa cells. Cryo-transmission electron microscopy showed that the structure of supramolecular assemblies BGTC:DOPE/β-gal and DOSP:MM27/K8 were different depending on liposome types and lipid/protein molar ratio. Finally, we observed that K8 treatment with DOSP:MM27/K8 rescues the cyclic adenosine monophosphate (cAMP-dependent chloride efflux in F508del-CFTR expressing cells, providing a new tool for the study of channelopathies.

  9. Vector-free intracellular delivery by reversible permeabilization.

    Directory of Open Access Journals (Sweden)

    Shirley O'Dea

    Full Text Available Despite advances in intracellular delivery technologies, efficient methods are still required that are vector-free, can address a wide range of cargo types and can be applied to cells that are difficult to transfect whilst maintaining cell viability. We have developed a novel vector-free method that uses reversible permeabilization to achieve rapid intracellular delivery of cargos with varying composition, properties and size. A permeabilizing delivery solution was developed that contains a low level of ethanol as the permeabilizing agent. Reversal of cell permeabilization is achieved by temporally and volumetrically controlling the contact of the target cells with this solution. Cells are seeded in conventional multi-well plates. Following removal of the supernatant, the cargo is mixed with the delivery solution and applied directly to the cells using an atomizer. After a short incubation period, permeabilization is halted by incubating the cells in a phosphate buffer saline solution that dilutes the ethanol and is non-toxic to the permeabilized cells. Normal culture medium is then added. The procedure lasts less than 5 min. With this method, proteins, mRNA, plasmid DNA and other molecules have been delivered to a variety of cell types, including primary cells, with low toxicity and cargo functionality has been confirmed in proof-of-principle studies. Co-delivery of different cargo types has also been demonstrated. Importantly, delivery occurs by diffusion directly into the cytoplasm in an endocytic-independent manner. Unlike some other vector-free methods, adherent cells are addressed in situ without the need for detachment from their substratum. The method has also been adapted to address suspension cells. This delivery method is gentle yet highly reproducible, compatible with high throughput and automated cell-based assays and has the potential to enable a broad range of research, drug discovery and clinical applications.

  10. Bioreducible Lipid-like Nanoparticles for Intracellular Protein Delivery

    Science.gov (United States)

    Arellano, Carlos Luis

    Protein-based therapy is one of the most direct ways to manipulate cell function and treat human disease. Although protein therapeutics has made its way to clinical practice, with five of the top fifteen global pharmaceuticals being peptide or protein-based drugs, one common limitation is that the effects of protein therapy are only achieved through the targeting of cell surface receptors and intracellular domains. Due to the impermeability of the cell membrane to most foreign materials, entire classes of potentially therapeutic proteins cannot thoroughly be studied without a safe and efficient method of transporting proteins into the cytosol. We report the use of a combinatorially-designed bioreducible lipid-like material (termed "lipidoid") - based protein delivery platform for the transfection of human cancer cell lines. Lipidoid nanoparticles are synthesized through a thin film dispersion method. The degradation of the bioreducible nanoparticles was observed when exposed to glutathione, a highly reductive compound present in the cytosol. We demonstrate that the nanoparticles are capable of transfecting a dose-dependent concentration of our model protein, beta-galactosidase into HeLa cells. Furthermore, formulations of the lipidoid containing the cytotoxic proteins saporin and RNase-A are both capable of inhibiting tumor cell proliferation as observed in in vitro treatment of different human cancer cell lines. There was no observed loss in protein activity after lyophilization and long--term storage, indicating the potential of pre-clinical applications. Overall, we demonstrate an effective approach to protein formulation and intracellular delivery. We believe that our formulations will lead to the study of a whole class of previously untapped therapeutics that may generate new solutions for previously untreatable diseases.

  11. Loperamide Restricts Intracellular Growth of Mycobacterium tuberculosis in Lung Macrophages.

    Science.gov (United States)

    Juárez, Esmeralda; Carranza, Claudia; Sánchez, Guadalupe; González, Mitzi; Chávez, Jaime; Sarabia, Carmen; Torres, Martha; Sada, Eduardo

    2016-12-01

    New approaches for improving tuberculosis (TB) control using adjunct host-directed cellular and repurposed drug therapies are needed. Autophagy plays a crucial role in the response to TB, and a variety of autophagy-inducing drugs that are currently available for various medical conditions may serve as an adjunct treatment in pulmonary TB. Here, we evaluated the potential of loperamide, carbamazepine, valproic acid, verapamil, and rapamycin to enhance the antimicrobial immune response to Mycobacterium tuberculosis (Mtb). Human monocyte-derived macrophages (MDMs) and murine alveolar cells (MACs) were infected with Mtb and treated with loperamide, carbamazepine, valproic acid, verapamil, and rapamycin in vitro. Balb/c mice were intraperitoneally administered loperamide, valproic acid, and verapamil, and MACs were infected in vitro with Mtb. The induction of autophagy, the containment of Mtb within autophagosomes and the intracellular Mtb burden were determined. Autophagy was induced by all of the drugs in human and mouse macrophages, and loperamide significantly increased the colocalization of microtubule-associated protein 1 light chain 3 with Mtb in MDMs. Carbamazepine, loperamide, and valproic acid induced microtubule-associated protein 1 light chain 3 and autophagy related 16- like protein 1 gene expression in MDMs and in MACs. Loperamide also induced a reduction in TNF-α production. Loperamide and verapamil induced autophagy, which was associated with a significant reduction in the intracellular growth of Mtb in MACs and alveolar macrophages. The intraperitoneal administration of loperamide and valproic acid induced autophagy in freshly isolated MACs. The antimycobacterial activity in MACs was higher after loperamide treatment and was associated with the degradation of p62. In conclusion, loperamide shows potential as an adjunctive therapy for the treatment of TB.

  12. Cannabidiol induces intracellular calcium elevation and cytotoxicity in oligodendrocytes.

    Science.gov (United States)

    Mato, Susana; Victoria Sánchez-Gómez, María; Matute, Carlos

    2010-11-01

    Heavy marijuana use has been linked to white matter histological alterations. However, the impact of cannabis constituents on oligodendroglial pathophysiology remains poorly understood. Here, we investigated the in vitro effects of cannabidiol, the main nonpsychoactive marijuana component, on oligodendrocytes. Exposure to cannabidiol induced an intracellular Ca(2+) rise in optic nerve oligodendrocytes that was not primarily mediated by entry from the extracellular space, nor by interactions with ryanodine or IP(3) receptors. Application of the mitochondrial protonophore carbonylcyanide-p-trifluoromethoxyphenylhydrazone (FCCP; 1 μM) completely prevented subsequent cannabidiol-induced Ca(2+) responses. Conversely, the increase in cytosolic Ca(2+) levels elicited by FCCP was reduced after previous exposure to cannabidiol, further suggesting that the mitochondria acts as the source of cannabidiol-evoked Ca(2+) rise in oligodendrocytes. n addition, brief exposure to cannabidiol (100 nM-10 μM) led to a concentration-dependent decrease of oligodendroglial viability that was not prevented by antagonists of CB(1), CB(2), vanilloid, A(2A) or PPARγ receptors, but was instead reduced in the absence of extracellular Ca(2+). The oligodendrotoxic effect of cannabidiol was partially blocked by inhibitors of caspase-3, -8 and -9, PARP-1 and calpains, suggesting the activation of caspase-dependent and -independent death pathways. Cannabidiol also elicited a concentration-dependent alteration of mitochondrial membrane potential, and an increase in reactive oxygen species (ROS) that was reduced in the absence of extracellular Ca(2+). Finally, cannabidiol-induced cytotoxicity was partially prevented by the ROS scavenger trolox. Together, these results suggest that cannabidiol causes intracellular Ca(2+) dysregulation which can lead to oligodendrocytes demise.

  13. Squalestatin alters the intracellular trafficking of a neurotoxic prion peptide

    Directory of Open Access Journals (Sweden)

    Williams Alun

    2007-11-01

    Full Text Available Abstract Background Neurotoxic peptides derived from the protease-resistant core of the prion protein are used to model the pathogenesis of prion diseases. The current study characterised the ingestion, internalization and intracellular trafficking of a neurotoxic peptide containing amino acids 105–132 of the murine prion protein (MoPrP105-132 in neuroblastoma cells and primary cortical neurons. Results Fluorescence microscopy and cell fractionation techniques showed that MoPrP105-132 co-localised with lipid raft markers (cholera toxin and caveolin-1 and trafficked intracellularly within lipid rafts. This trafficking followed a non-classical endosomal pathway delivering peptide to the Golgi and ER, avoiding classical endosomal trafficking via early endosomes to lysosomes. Fluorescence resonance energy transfer analysis demonstrated close interactions of MoPrP105-132 with cytoplasmic phospholipase A2 (cPLA2 and cyclo-oxygenase-1 (COX-1, enzymes implicated in the neurotoxicity of prions. Treatment with squalestatin reduced neuronal cholesterol levels and caused the redistribution of MoPrP105-132 out of lipid rafts. In squalestatin-treated cells, MoPrP105-132 was rerouted away from the Golgi/ER into degradative lysosomes. Squalestatin treatment also reduced the association between MoPrP105-132 and cPLA2/COX-1. Conclusion As the observed shift in peptide trafficking was accompanied by increased cell survival these studies suggest that the neurotoxicity of this PrP peptide is dependent on trafficking to specific organelles where it activates specific signal transduction pathways.

  14. Strategies for Retaining Highly Qualified & Experienced Technical Teachers in Teaching Profession in Katsina State, Nigeria

    OpenAIRE

    Saifullahi Kasim Tafida; Che Kum Clement; Md. Abu Raihan

    2015-01-01

    The objective of this study is to identify the strategies for retaining highly qualified & experienced technical teachers of technical and vocational educational institutions in Katsina State of Nigeria. Two research questions were formulated to guide the study. A 26 items survey questionnaire was developed and used to elicit responses from technical education administrators, technical education principals/vice principals and technical teachers in technical and vocational education institutio...

  15. Air Force Officer Qualifying Test Form T: Initial Item-, Test-, Factor-, and Composite-Level Analyses

    Science.gov (United States)

    2016-12-01

    used to qualify applicants for ROTC and OTS officer commissioning programs. The Pilot, Combat Systems Officer (CSO), and Air Battle Manager ( ABM ...AIR FORCE RESEARCH LABORATORY 711 HUMAN PERFORMANCE WING, AIRMAN SYSTEMS DIRECTORATE, WRIGHT-PATTERSON AIR FORCE BASE, OH 45433 AIR FORCE MATERIEL...Warfighter Interface Division Airman Systems Directorate This report is published in the interest of scientific and technical information exchange, and

  16. From space qualified fiber optic gyroscope to generic fiber optic solutions available for space application

    Science.gov (United States)

    Buret, Thomas; Ramecourt, David; Napolitano, Fabien

    2017-11-01

    The aim of this article is to present how the qualification of the Fiber Optic Gyroscope technology from IXSEA has been achieved through the qualification of a large range of optical devices and related manufacturing processes. These qualified optical devices and processes, that are now fully mastered by IXSEA through vertical integration of the technology, can be used for other space optical sensors. The example of the SWARM project will be discussed.

  17. Marriage and gender inequality: work–family arrangements of Portuguese and British highly qualified women

    OpenAIRE

    Rosa, Rodrigo; Lorga, Catarina; Lyonette, Clare

    2016-01-01

    This article examines work–family reconciliation processes in order to understand if, over the course of marital life, women become socially closer or further away from their partner. Drawing on work–life interviews with highly qualified women in Portugal and Britain, we compare these processes in two societies with different historical and social backgrounds. Findings reveal three main configurations of social (in)equality which emerge during married life: growing inequality i...

  18. Nutritional status assessment in colorectal cancer patients qualified to systemic treatment

    OpenAIRE

    Ziętarska, Monika; Krawczyk-Lipiec, Joanna; Kraj, Leszek; Zaucha, Renata; Małgorzewicz, Sylwia

    2017-01-01

    Aim of the study Cancer is usually associated with impaired nutritional status, which is one of the factors contributing to the deterioration of the results of surgery, chemotherapy, or radiotherapy. The aim of this study was the assessment of the nutritional status of patients with CRC qualified to chemotherapy. Material and methods Seventy-five persons aged 40–86 years with colorectal cancer were examined. To evaluate the nutritional status NRS 2002, SGA, SCRINIO Working Group classificatio...

  19. Nutritional status assessment in colorectal cancer patients qualified to systemic treatment

    OpenAIRE

    Monika Ziętarska; Joanna Krawczyk-Lipiec; Leszek Kraj; Renata Zaucha; Sylwia Małgorzewicz

    2017-01-01

    Aim of the study . Cancer is usually associated with impaired nutritional status, which is one of the factors contributing to the deterioration of the results of surgery, chemotherapy, or radiotherapy. The aim of this study was the assessment of the nutritional status of patients with CRC qualified to chemotherapy. Material and methods : Seventy-five persons aged 40–86 years with colorectal cancer were examined. To evaluate the nutritional status NRS 2002, SGA, SCRINIO Working Group...

  20. Experience with neutron flux monitoring systems qualified for post-accident monitoring

    International Nuclear Information System (INIS)

    Shugars, H.G.; Miller, J.F.

    1995-01-01

    In this paper we discuss the environmental requirements for excore neutron flux monitors that are qualified for use during and after postulated accidents in Pressurized Water Reactors (PWRs). We emphasize PWRs designed in the United States, which are similar to those used also in parts of Western Europe and Eastern Asia. We then discuss design features of the flux monitoring systems necessary to address the environmental, functional, and regulatory requirements, and the experience with these systems. (author). 9 refs, 2 figs

  1. Application of a qualified RETRAN model to plant transient evaluation support

    International Nuclear Information System (INIS)

    Sedano, P.G.; Mata, P.; Alcantud, F.; Serra, J.; Castrillo, F.

    1989-01-01

    This paper presents the applicability and usefulness of a complete and well qualified plant transient code and model to support in depth evaluation of anomalous plant transients. Analyses of several operational and abnormal transients that ocurred during the first three cycles of Cofrentes (BWR-6) NPP are presented. This application demonstrated the need of a very detailed and adjusted simulation of the control systems as well as the convenience of having as complete as possible data adquisition system. (orig.)

  2. Application of a qualified RETRAN model to plant transient evaluation support

    International Nuclear Information System (INIS)

    Sedano, P.G.; Mata, P.; Alcantud, F.; Serra, J.

    1989-01-01

    This paper presents the applicability and usefulness of a complete and well qualified plant transient code and model to support in depth evaluation of anomalous plant transients. Analyses of several operational and abnormal transients occurred during the first three cycles of Cofrentes (BWR-6) NPP are presented. This application remarked the need of a very detailed and adjusted simulation of the control systems as well as the convenience of having an as complete as possible data acquisition system

  3. Capacity market design and renewable energy: Performance incentives, qualifying capacity, and demand curves

    Energy Technology Data Exchange (ETDEWEB)

    Botterud, Audun; Levin, Todd; Byers, Conleigh

    2018-01-01

    A review of capacity markets in the United States in the context of increasing levels of variable renewable energy finds substantial differences with respect to incentives for operational performance, methods to calculate qualifying capacity for variable renewable energy and energy storage, and demand curves for capacity. The review also reveals large differences in historical capacity market clearing prices. The authors conclude that electricity market design must continue to evolve to achieve cost-effective policies for resource adequacy.

  4. THE UNQUALIFIED MEDICAL PRACTITIONERS - Methods of Practice and Nexus with the Qualified Doctors

    OpenAIRE

    K.V. Narayana

    2006-01-01

    The private sector accounts for about 75 percent of outpatient as well as inpatient medical care in Andhra Pradesh. The presence of a large number of unqualified medical practitioners in the rural areas and urban slums indicate that they provide most of the outpatient services in the private sector. Given the huge quantum of services provided by the RMPs, the present study aims at identifying their number, characteristics and the nexus with the qualified doctors through a case study of one di...

  5. The research of tactics of the competitive activity of the qualified sportswomen in long jumps

    OpenAIRE

    Grebenyuk, Oleg

    2016-01-01

    Grebenyuk A. Purpose: to research features of competitive tactics of the qualified sportswomen in long jumps from the running start. Material & Methods: such methods were applied the in researches: analysis of scientific and methodical literature, pedagogical supervision, content analysis (analysis of protocols of competitions). Results of 152 jumpers in length, who executed 886 attempts, are analyzed for the solution of the stated purpose. Results: options of competitive tactics of the q...

  6. Nuclear magnetic resonance studies of intracellular ions in perfused from heart

    International Nuclear Information System (INIS)

    Burnstein, D.; Fossel, E.T.

    1987-01-01

    Intracellular sodium, potassium, and lithium were observed in a perfused frog heart by nuclear magnetic resonance (NMR) spectroscopy. A perfusate buffer containing the shift reagent, dysprosium tripolyphosphate, was used in combination with mathematical filtering or presaturation of the extracellular resonance to separate the intra- and extracellular sodium NMR signals. Addition of 10 μM ouabain to the perfusate, perfusion with a zero potassium, low-calcium buffer, and replacement of 66% of the perfusate sodium with lithium resulted in changes in the intracellular sodium levels. An increase of 45% in the intracellular sodium was observed when changing the pacing rate from 0 to 60 beats/min (with proportional changes for intermediate pacing rates). The ratio of intracellular potassium to sodium concentration was determined to be 2.3 by NMR, indicating that a substantial amount of the intracellular potassium is undetectable with these NMR method. In addition, intracellular lithium was observed during perfusion with a lithium-containing perfusate

  7. Can Social Robots Qualify for Moral Consideration? Reframing the Question about Robot Rights

    Directory of Open Access Journals (Sweden)

    Herman T. Tavani

    2018-03-01

    Full Text Available A controversial question that has been hotly debated in the emerging field of robot ethics is whether robots should be granted rights. Yet, a review of the recent literature in that field suggests that this seemingly straightforward question is far from clear and unambiguous. For example, those who favor granting rights to robots have not always been clear as to which kinds of robots should (or should not be eligible; nor have they been consistent with regard to which kinds of rights—civil, legal, moral, etc.—should be granted to qualifying robots. Also, there has been considerable disagreement about which essential criterion, or cluster of criteria, a robot would need to satisfy to be eligible for rights, and there is ongoing disagreement as to whether a robot must satisfy the conditions for (moral agency to qualify either for rights or (at least some level of moral consideration. One aim of this paper is to show how the current debate about whether to grant rights to robots would benefit from an analysis and clarification of some key concepts and assumptions underlying that question. My principal objective, however, is to show why we should reframe that question by asking instead whether some kinds of social robots qualify for moral consideration as moral patients. In arguing that the answer to this question is “yes,” I draw from some insights in the writings of Hans Jonas to defend my position.

  8. APPROACH TO SYNTHESIS OF PASSIVE INFRARED DETECTORS BASED ON QUASI-POINT MODEL OF QUALIFIED INTRUDER

    Directory of Open Access Journals (Sweden)

    I. V. Bilizhenko

    2017-01-01

    Full Text Available Subject of Research. The paper deals with synthesis of passive infra red (PIR detectors with enhanced detection capability of qualified intruder who uses different types of detection countermeasures: the choice of specific movement direction and disguise in infrared band. Methods. We propose an approach based on quasi-point model of qualified intruder. It includes: separation of model priority parameters, formation of partial detection patterns adapted to those parameters and multi channel signal processing. Main Results. Quasi-pointmodel of qualified intruder consisting of different fragments was suggested. Power density difference was used for model parameters estimation. Criteria were formulated for detection pattern parameters choice on the basis of model parameters. Pyroelectric sensor with nine sensitive elements was applied for increasing the signal information content. Multi-channel processing with multiple partial detection patterns was proposed optimized for detection of intruder's specific movement direction. Practical Relevance. Developed functional device diagram can be realized both by hardware and software and is applicable as one of detection channels for dual technology passive infrared and microwave detectors.

  9. Towards Qualifiable Code Generation from a Clocked Synchronous Subset of Modelica

    Directory of Open Access Journals (Sweden)

    Bernhard Thiele

    2015-01-01

    Full Text Available So far no qualifiable automatic code generators (ACGs are available for Modelica. Hence, digital control applications can be modeled and simulated in Modelica, but require tedious additional efforts (e.g., manual reprogramming to produce qualifiable target system production code. In order to more fully leverage the potential of a model-based development (MBD process in Modelica, a qualifiable automatic code generator is needed. Typical Modelica code generation is a fairly complex process which imposes a huge development burden to any efforts of tool qualification. This work aims at mapping a Modelica subset for digital control function development to a well-understood synchronous data-flow kernel language. This kernel language allows to resort to established compilation techniques for data-flow languages which are understood enough to be accepted by certification authorities. The mapping is established by providing a translational semantics from the Modelica subset to the synchronous data-flow kernel language. However, this translation turned out to be more intricate than initially expected and has given rise to several interesting issues that require suitable design decisions regarding the mapping and the language subset.

  10. TopWorx/GO switch new generation nuclear qualified proximity position sensors

    International Nuclear Information System (INIS)

    Merrifield, G.

    2011-01-01

    An overview of the benefits of installing TopWorx/GO Nuclear Qualified Proximity Position Sensors instead of traditional mechanical switches. Mechanical switches have been the standard for the nuclear industry for years, but that is only because of lack of competition. Because of multiple moving parts, three-piece design, and new low current control systems, mechanical switches are susceptible to a host of environmental factors that cause them to break and/or fail. TopWorx/GO Switch is a smaller, rugged, dependable, one-piece Stainless Steel proximity position sensor that that exceeds CANDU and Global Qualification Levels, easily replacing mechanical switches. It is the only position sensor to meet or exceed Westinghouse AP 1000 specifications as well. Nuclear facilities save money in maintenance reductions, extended PM's and reduction in 'Man REM' hours. This is achieved by not having to service and replace GO Switch position sensors as often as mechanical switches, since their qualified life is 100 years +1 Post Accident submerged. Plant safety is increased due to less switch failure, higher qualification testing levels, repeatability and longer qualified life. (author)

  11. HIGHLY QUALIFIED WORKING FORCE – KEY ELEMENT OF INNOVATIVE DEVELOPMENT MODEL

    Directory of Open Access Journals (Sweden)

    M. Avksientiev

    2014-12-01

    Full Text Available Highly qualified working force is a central element of intensive development model in modern society. The article surveys the experience of countries that managed to transform their economy to the innovative one. Ukrainian economy cannot stand aside processes that dominate the world economy trends, thus we are to use this experience to succeed in future. Today any government of the world is facing challenges that occur due to transformation of the economy into informational one. This type of economy causes its transformation form extensive to intensive one. The main reasons under that is limitation of nature resources, material factors of production. Thus this approach depends much on the quality of working force. Unfortunately in Ukraine there is a misbalance in specialist preparation. This puts additional pressure on the educational sphere also. In order to avoid this pressure we are to conduct reforms in education sphere. Nowadays, in the world views and concepts of governmental role in the social development are changing. This why, even at times of economic recession educational costs are not reduced under the new economical doctrine in the EU. Highly qualified specialists, while creating new products and services play role of engineers in XXI century. They are to lead their industries to world leading positions. From economic point of view, highly qualified specialists benefit society with higher income rates, taxation and thus, increasing the living standards in society. Thus, the majority if modern scientists prove the importance of highly trained working force for more effective economic development.

  12. A Thapsigargin-Resistant Intracellular Calcium Sequestering Compartment in Rat Brain

    Science.gov (United States)

    2000-03-31

    have a major impact on neuronal intracellular signaling. Most of the ER in neurons and glia appears to accumulate calcium by energy driven ion pumps...secretion of exocrine, endocrine, and neurocrine products, regulation of glycogenolysis and gluconeogenesis , intracellular transport, secretion of fluids...the RyRs [140]. Furthermore, the intracellular expression of these receptor-channels in neuronal ER is also reciprocal with RyRs located primarily in

  13. Qualified Coatings in Nuclear Power Plants. Commercial products; Qualified Coatings in Nuclear Power Plants. Commercial products. Pinturas homologadas en centrales nucleares. Productos comerciales

    Energy Technology Data Exchange (ETDEWEB)

    Barcena, J.; Nunez, B.; Romero, M.; Baladiam, M.

    2014-07-01

    Recently, the supplier of paints that were qualified for use in nuclear applications as protective coatings have ceased to supply in Spain the paints that was used in areas or components with special requirements for nuclear power plants (NPPs). This lack of the common commercial products called for the search for and homologation of other products. A study was performed on the current status of the homologation of commercial products for NPPs and on the codes and standards governing them. The criteria to be met have been defined and the results of the tests performed on the selected paints have been compared against the established criteria so as to allow the homologation of the paints. (Author)

  14. Influence of intracellular acidosis on contractile function in the working rat heart

    International Nuclear Information System (INIS)

    Jeffrey, F.M.H.; Malloy, C.R.; Radda, G.K.

    1987-01-01

    The decrease in myocardial contractility during ischemia, hypoxia, and extracellular acidosis has been attributed to intracellular acidosis. Previous studies of the relationship between pH and contractile state have utilized respiratory or metabolic acidosis to alter intracellular pH. The authors developed a model in the working perfused rat heart to study the effects of intracellular acidosis with normal external pH and optimal O 2 delivery. Intracellular pH and high-energy phosphates were monitored by 31 P nuclear magnetic resonance spectroscopy. Hearts were perfused to a steady state with a medium containing 10 mM NH 4 Cl. Acidosis induced a substantial decrease in aortic flow and stroke volume which was associated with little change in peak systolic pressure. It was concluded that (1) for the same intracellular acidosis the influence on tension development was more pronounced with a combined extra- and intracellular acidosis than with an isolated intracellular acidosis, and (2) stroke volume at constant preload was impaired by intracellular acidosis even though changes in developed pressure were minimal. These observations suggest that isolated intracellular acidosis has adverse effects on diastolic compliance and/or relaxation

  15. Subversion of the cytoskeleton by intracellular bacteria: lessons from Listeria, Salmonella, and Vibrio

    Science.gov (United States)

    de Souza Santos, Marcela; Orth, Kim

    2018-01-01

    Summary Entry into host cells and intracellular persistence by invasive bacteria are tightly coupled to the ability of the bacterium to disrupt the eukaryotic cytoskeletal machinery. Herein we review the main strategies used by three intracellular pathogens to harness key modulators of the cytoskeleton. Two of these bacteria, namely Listeria monocytogenes and Salmonella enterica serovar Typhimurium, exhibit quite distinct intracellular lifestyles, and therefore, provide a comprehensive panel for the understanding of the intricate bacteria-cytoskeleton interplay during infections. The emerging intracellular pathogen Vibrio parahaemolyticus is depicted as a developing model for the uncovering of novel mechanisms used to hijack the cytoskeleton. PMID:25440316

  16. Cationic Antimicrobial Peptide LL-37 Is Effective against both Extra- and Intracellular Staphylococcus aureus

    Science.gov (United States)

    Noore, Jabeen; Noore, Adly

    2013-01-01

    The increasing resistance of bacteria to conventional antibiotics and the challenges posed by intracellular bacteria, which may be responsible for chronic and recurrent infections, have driven the need for advanced antimicrobial drugs for effective elimination of both extra- and intracellular pathogens. The purpose of this study was to determine the killing efficacy of cationic antimicrobial peptide LL-37 compared to conventional antibiotics against extra- and intracellular Staphylococcus aureus. Bacterial killing assays and an infection model of osteoblasts and S. aureus were studied to determine the bacterial killing efficacy of LL-37 and conventional antibiotics against extra- and intracellular S. aureus. We found that LL-37 was effective in killing extracellular S. aureus at nanomolar concentrations, while lactoferricin B was effective at micromolar concentrations and doxycycline and cefazolin at millimolar concentrations. LL-37 was surprisingly more effective in killing the clinical strain than in killing an ATCC strain of S. aureus. Moreover, LL-37 was superior to conventional antibiotics in eliminating intracellular S. aureus. The kinetic studies further revealed that LL-37 was fast in eliminating both extra- and intracellular S. aureus. Therefore, LL-37 was shown to be very potent and prompt in eliminating both extra- and intracellular S. aureus and was more effective in killing extra- and intracellular S. aureus than commonly used conventional antibiotics. LL-37 could potentially be used to treat chronic and recurrent infections due to its effectiveness in eliminating not only extracellular but also intracellular pathogens. PMID:23274662

  17. Neutrophilic granulocytes reactive response in candida vulvovaginitis patients with intracellular microorganism persistence complications

    OpenAIRE

    YAKOVYCHUK NINA DMYTRIVNA; DJUIRIAK VALENTYNA STEPANIVNA

    2015-01-01

    Polymorphic neutrophilic granulocytes reactive response and body immune reactivity in general considerably decrease in patients suffering from candida vaginitis on the basis of intracellular microorganisms persistence.

  18. Changes in sickness absenteeism following the introduction of a qualifying day for sickness benefit--findings from Sweden Post

    DEFF Research Database (Denmark)

    Voss, M; Floderus, B; Diderichsen, F

    2001-01-01

    AIMS: In 1993, a qualifying day without sickness benefit was introduced to the Swedish sickness benefit system. The aim of the present study is to investigate sickness absenteeism before and after the introduction of the qualifying day, in the light of conditions inside and outside working life....... METHODS: The study was based on 1,952 female and 2,229 male employees of Sweden Post. Sickness absence was measured by sickness incidence one year before and one year after the introduction of the qualifying day (sick-leave events/person days at risk). Information about explanatory factors was collected...

  19. 26 CFR 1.188-1 - Amortization of certain expenditures for qualified on-the-job training and child care facilities.

    Science.gov (United States)

    2010-04-01

    ... qualified on-the-job training and child care facilities. 1.188-1 Section 1.188-1 Internal Revenue INTERNAL... qualified on-the-job training and child care facilities. (a) Allowance of deduction—(1) In general. Under... operation of a qualified on-the-job training or child care facility or are integrally related facilities...

  20. Comparison of the intracellular trafficking itinerary of ctla-4 orthologues.

    Directory of Open Access Journals (Sweden)

    Satdip Kaur

    Full Text Available CTLA-4 is an essential inhibitor of T cell immune responses. At steady state, most CTLA-4 resides in intracellular compartments due to constitutive internalisation mediated via a tyrosine based endocytic motif (YVKM within the cytoplasmic domain. This domain is highly conserved in mammals suggesting strong selective pressure. In contrast, the C-terminal domain varies considerably in non-mammals such as fish, xenopus and birds. We compared the ability of the C-terminus of these species to direct the trafficking of CTLA-4 with human CTLA-4. Using a chimeric approach, endocytosis was found to be conserved between human, xenopus and chicken CTLA-4 but was reduced substantially in trout CTLA-4, which lacks the conserved YXXM motif. Nevertheless, we identified an alternative YXXF motif in trout CTLA-4 that permitted limited endocytosis. Post-internalisation, CTLA-4 was either recycled or targeted for degradation. Human and chicken CTLA-4, which contain a YVKM motif, showed efficient recycling compared to xenopus CTLA-4 which contains a less efficient YEKM motif. Specific mutation of this motif in human CTLA-4 reduced receptor recycling. These findings suggest evolutionary development in the endocytic and recycling potential of CTLA-4, which may facilitate more refined functions of CTLA-4 within the mammalian immune system.

  1. Transepithelial Na+ transport and the intracellular fluids: a computer study.

    Science.gov (United States)

    Civan, M M; Bookman, R J

    1982-01-01

    Computer simulations of tight epithelia under three experimental conditions have been carried out, using the rheogenic nonlinear model of Lew, Ferreira and Moura (Proc. Roy. Soc. London. B 206:53-83, 1979) based largely on the formulation of Koefoed-Johnsen and Ussing (Acta Physiol. Scand. 42: 298-308. 1958). First, analysis of the transition between the short-circuited and open-circuited states has indicated that (i) apical Cl- permeability is a critical parameter requiring experimental definition in order to analyze cell volume regulation, and (ii) contrary to certain experimental reports, intracellular Na+ concentration (ccNa) is expected to be a strong function of transepithelial clamping voltage. Second, analysis of the effects of lowering serosal K+ concentration (csK) indicates that the basic model cannot simulate several well-documented observations; these defects can be overcome, at least qualitatively, by modifying the model to take account of the negative feedback interaction likely to exist between the apical Na+ permeability and ccNa. Third, analysis of the strongly supports the concept that osmotically induced permeability changes in the apical intercellular junctions play a physiological role in conserving the body's stores of NaCl. The analyses also demonstrate that the importance of Na+ entry across the basolateral membrane is strongly dependent upon transepithelial potential, cmNa and csK; under certain conditions, net Na+ entry could be appreciably greater across the basolateral than across the apical membrane.

  2. Viral evasion of intracellular DNA and RNA sensing

    Science.gov (United States)

    Chan, Ying Kai; Gack, Michaela U.

    2016-01-01

    The co-evolution of viruses with their hosts has led to the emergence of viral pathogens that are adept at evading or actively suppressing host immunity. Pattern recognition receptors (PRRs) are key components of antiviral immunity that detect conserved molecular features of viral pathogens and initiate signalling that results in the expression of antiviral genes. In this Review, we discuss the strategies that viruses use to escape immune surveillance by key intracellular sensors of viral RNA or DNA, with a focus on RIG-I-like receptors (RLRs), cyclic GMP–AMP synthase (cGAS) and interferon-γ (IFNγ)-inducible protein 16 (IFI16). Such viral strategies include the sequestration or modification of viral nucleic acids, interference with specific post-translational modifications of PRRs or their adaptor proteins, the degradation or cleavage of PRRs or their adaptors, and the sequestration or relocalization of PRRs. An understanding of viral immune-evasion mechanisms at the molecular level may guide the development of vaccines and antivirals. PMID:27174148

  3. Sigma-1 receptor: The novel intracellular target of neuropsychotherapeutic drugs

    Directory of Open Access Journals (Sweden)

    Teruo Hayashi

    2015-01-01

    Full Text Available Sigma-1 receptor ligands have been long expected to serve as drugs for treatment of human diseases such as neurodegenerative disorders, depression, idiopathic pain, drug abuse, and cancer. Recent research exploring the molecular function of the sigma-1 receptor started unveiling underlying mechanisms of the therapeutic activity of those ligands. Via the molecular chaperone activity, the sigma-1 receptor regulates protein folding/degradation, ER/oxidative stress, and cell survival. The chaperone activity is activated or inhibited by synthetic sigma-1 receptor ligands in an agonist-antagonist manner. Sigma-1 receptors are localized at the endoplasmic reticulum (ER membranes that are physically associated with the mitochondria (MAM: mitochondria-associated ER membrane. In specific types of neurons (e.g., those at the spinal cord, sigma-1 receptors are also clustered at ER membranes that juxtapose postsynaptic plasma membranes. Recent studies indicate that sigma-1 receptors, partly in sake of its unique subcellular localization, regulate the mitochondria function that involves bioenergetics and free radical generation. The sigma-1 receptor may thus provide an intracellular drug target that enables controlling ER stress and free radical generation under pathological conditions.

  4. Evaluating nanoparticle sensor design for intracellular pH measurements.

    Science.gov (United States)

    Benjaminsen, Rikke V; Sun, Honghao; Henriksen, Jonas R; Christensen, Nynne M; Almdal, Kristoffer; Andresen, Thomas L

    2011-07-26

    Particle-based nanosensors have over the past decade been designed for optical fluorescent-based ratiometric measurements of pH in living cells. However, quantitative and time-resolved intracellular measurements of pH in endosomes and lysosomes using particle nanosensors are challenging, and there is a need to improve measurement methodology. In the present paper, we have successfully carried out time-resolved pH measurements in endosomes and lyosomes in living cells using nanoparticle sensors and show the importance of sensor choice for successful quantification. We have studied two nanoparticle-based sensor systems that are internalized by endocytosis and elucidated important factors in nanosensor design that should be considered in future development of new sensors. From our experiments it is clear that it is highly important to use sensors that have a broad measurement range, as erroneous quantification of pH is an unfortunate result when measuring pH too close to the limit of the sensitive range of the sensors. Triple-labeled nanosensors with a pH measurement range of 3.2-7.0, which was synthesized by adding two pH-sensitive fluorophores with different pK(a) to each sensor, seem to be a solution to some of the earlier problems found when measuring pH in the endosome-lysosome pathway.

  5. [Hyperosmolarity: Intracellular effects and implication in dry eye disease].

    Science.gov (United States)

    Warcoin, E; Clouzeau, C; Brignole-Baudouin, F; Baudouin, C

    2016-09-01

    Dry eye disease is a multifactorial disease affecting the lacrimal functional unit and which has a significant impact on the quality of life of patients. This pathology works as a vicious circle at the ocular surface in which hyperosmolarity of the tear film plays a key role. This review intends to describe the different reported intracellular effects induced by hyperosmolarity in cells: alteration of cytoskeleton, cell cycle slowdown, adaptation mechanisms triggered as restoration of cell volume and accumulation of compatible osmolytes, the crucial role of the osmoprotectant factor Nuclear Factor of the Activated T cells-5 (NFAT5), apoptosis, as well as oxidative stress and inflammatory responses caused by this particular condition. Reported effects of hyperosmolarity in the experimental studies specific of dry eye disease concerning ocular surface cells will be described in parallel. Indeed, these data allow to understand a part of the pathophysiology of the disease, and specially the links between tear hyperosmolarity and inflammation of the ocular surface, the second key of the pathology phenomenon. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  6. Intracellular magnetophoresis of amyloplasts and induction of root curvature

    Science.gov (United States)

    Kuznetsov, O. A.; Hasenstein, K. H.

    1996-01-01

    High-gradient magnetic fields (HGMFs) were used to induce intracellular magnetophoresis of amyloplasts. The HGMFs were generated by placing a small ferromagnetic wedge into a uniform magnetic field or at the gap edge between two permanent magnets. In the vicinity of the tip of the wedge the dynamic factor of the magnetic field, delta(H2/2), was about 10(9) Oe2.cm-1, which subjected the amyloplasts to a force comparable to that of gravity. When roots of 2-d-old seedlings of flax (Linum usitatissimum L.) were positioned vertically and exposed to an HGMF, curvature away from the wedge was transient and lasted approximately 1 h. Average curvature obtained after placing magnets, wedge and seedlings on a 1-rpm clinostat for 2 h was 33 +/- 5 degrees. Roots of horizontally placed control seedlings without rotation curved about 47 +/- 4 degrees. The time course of curvature and changes in growth rate were similar for gravicurvature and for root curvature induced by HGMFs. Microscopy showed displacement of amyloplasts in vitro and in vivo. Studies with Arabidopsis thaliana (L.) Heynh. showed that the wild type responded to HGMFs but the starchless mutant TC7 did not. The data indicate that a magnetic force can be used to study the gravisensing and response system of roots.

  7. Post-translational Control of Intracellular Pathogen Sensing Pathways.

    Science.gov (United States)

    Chiang, Cindy; Gack, Michaela U

    2017-01-01

    Mammalian cells recognize virus-derived nucleic acids using a defined set of intracellular sensors including the DNA sensors cyclic GMP-AMP (cGAMP) synthase (cGAS) and interferon gamma (IFNγ)-inducible protein 16 (IFI16) as well as viral RNA receptors of the retinoic acid-inducible gene I (RIG-I)-like receptor (RLR) family. Following innate immune recognition, these sensors launch an immune response that is characterized by the transcriptional upregulation of many antiviral molecules, including proinflammatory cytokines, chemokines, and IFN-stimulated genes. Recent studies have demonstrated that the signal transduction initiated by these sensors is sophisticatedly regulated by post-translational modifications (PTMs) resulting in a robust yet 'tunable' cytokine response to maintain immune homeostasis. Here we summarize recent advances in our understanding of how PTMs and regulatory enzymes control the signaling activity of RLRs, cGAS, and IFI16 as well as their proximal adaptor proteins. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Iron in intracellular infection: to provide or to deprive?

    Directory of Open Access Journals (Sweden)

    Sandro eSilva-Gomes

    2013-12-01

    Full Text Available Due to their chemical versatility, transition metals were incorporated as cofactors for several basic metabolic pathways in living organisms. This same characteristic makes them potentially harmful, since they can be engaged in deleterious reactions like Fenton chemistry. As such, organisms have evolved highly specialized mechanisms to supply their own metal needs while keeping their toxic potential in check.This dual character comes into play in host-pathogen interactions, given that the host can either deprive the pathogen of these key nutrients or exploit them to induce toxicity towards the invading agent. Iron stands as the prototypic example of how a metal can be used to limit the growth of pathogens by nutrient deprivation, a mechanism widely studied in Mycobacterium infections. However, the host can also take advantage of iron-induced toxicity to control pathogen proliferation, as observed in infections caused by Leishmania. Whether we may harness either of the two pathways for therapeutical purposes is still ill-defined.In this review, we discuss how modulation of the host iron availability impacts the course of infections, focusing on those caused by two relevant intracellular pathogens, Mycobacterium and Leishmania.

  9. Into the intracellular logistics of cross-presentation

    Directory of Open Access Journals (Sweden)

    Charlotte eSadaka

    2012-02-01

    Full Text Available The induction of cytotoxic CD8+ T cell responses requires the presentation of antigenic peptides by MHC class I molecules (MHC I. MHC I usually present peptides derived from endogenous proteins. However, some subtypes of dendritic cells (DCs have developed the ability to efficiently present peptides derived from exogenous antigens on MHC I via a process called cross-presentation. Cross-presentation is intimately linked to the induction of anti-viral, -bacterial and -tumor cytotoxic T cell (CTL responses, as well as a wide variety of CTL-mediated diseases and transplant rejections. The molecular and cellular mechanisms underlying cross-presentation have been studied intensively since its original description, yet understanding of this process is incomplete and on the forefront of immunological research. Numerous pathways and models, some of them conflicting, have been described so far. Here, we review the various pathways reported as involved in cross-presentation, highlighting the complexity of this process. We also discuss in detail the different intracellular steps required, from antigen capture and routing, to processing and finally peptide loading, emphasizing the need for a better understanding of the cell biology of this phenomenon.

  10. The Chlamydia psittaci genome: a comparative analysis of intracellular pathogens.

    Science.gov (United States)

    Voigt, Anja; Schöfl, Gerhard; Saluz, Hans Peter

    2012-01-01

    Chlamydiaceae are a family of obligate intracellular pathogens causing a wide range of diseases in animals and humans, and facing unique evolutionary constraints not encountered by free-living prokaryotes. To investigate genomic aspects of infection, virulence and host preference we have sequenced Chlamydia psittaci, the pathogenic agent of ornithosis. A comparison of the genome of the avian Chlamydia psittaci isolate 6BC with the genomes of other chlamydial species, C. trachomatis, C. muridarum, C. pneumoniae, C. abortus, C. felis and C. caviae, revealed a high level of sequence conservation and synteny across taxa, with the major exception of the human pathogen C. trachomatis. Important differences manifest in the polymorphic membrane protein family specific for the Chlamydiae and in the highly variable chlamydial plasticity zone. We identified a number of psittaci-specific polymorphic membrane proteins of the G family that may be related to differences in host-range and/or virulence as compared to closely related Chlamydiaceae. We calculated non-synonymous to synonymous substitution rate ratios for pairs of orthologous genes to identify putative targets of adaptive evolution and predicted type III secreted effector proteins. This study is the first detailed analysis of the Chlamydia psittaci genome sequence. It provides insights in the genome architecture of C. psittaci and proposes a number of novel candidate genes mostly of yet unknown function that may be important for pathogen-host interactions.

  11. The mechanical environment modulates intracellular calcium oscillation activities of myofibroblasts.

    Directory of Open Access Journals (Sweden)

    Charles Godbout

    Full Text Available Myofibroblast contraction is fundamental in the excessive tissue remodeling that is characteristic of fibrotic tissue contractures. Tissue remodeling during development of fibrosis leads to gradually increasing stiffness of the extracellular matrix. We propose that this increased stiffness positively feeds back on the contractile activities of myofibroblasts. We have previously shown that cycles of contraction directly correlate with periodic intracellular calcium oscillations in cultured myofibroblasts. We analyze cytosolic calcium dynamics using fluorescent calcium indicators to evaluate the possible impact of mechanical stress on myofibroblast contractile activity. To modulate extracellular mechanics, we seeded primary rat subcutaneous myofibroblasts on silicone substrates and into collagen gels of different elastic modulus. We modulated cell stress by cell growth on differently adhesive culture substrates, by restricting cell spreading area on micro-printed adhesive islands, and depolymerizing actin with Cytochalasin D. In general, calcium oscillation frequencies in myofibroblasts increased with increasing mechanical challenge. These results provide new insight on how changing mechanical conditions for myofibroblasts are encoded in calcium oscillations and possibly explain how reparative cells adapt their contractile behavior to the stresses occurring in normal and pathological tissue repair.

  12. Distributed and dynamic intracellular organization of extracellular information.

    Science.gov (United States)

    Granados, Alejandro A; Pietsch, Julian M J; Cepeda-Humerez, Sarah A; Farquhar, Iseabail L; Tkačik, Gašper; Swain, Peter S

    2018-06-05

    Although cells respond specifically to environments, how environmental identity is encoded intracellularly is not understood. Here, we study this organization of information in budding yeast by estimating the mutual information between environmental transitions and the dynamics of nuclear translocation for 10 transcription factors. Our method of estimation is general, scalable, and based on decoding from single cells. The dynamics of the transcription factors are necessary to encode the highest amounts of extracellular information, and we show that information is transduced through two channels: Generalists (Msn2/4, Tod6 and Dot6, Maf1, and Sfp1) can encode the nature of multiple stresses, but only if stress is high; specialists (Hog1, Yap1, and Mig1/2) encode one particular stress, but do so more quickly and for a wider range of magnitudes. In particular, Dot6 encodes almost as much information as Msn2, the master regulator of the environmental stress response. Each transcription factor reports differently, and it is only their collective behavior that distinguishes between multiple environmental states. Changes in the dynamics of the localization of transcription factors thus constitute a precise, distributed internal representation of extracellular change. We predict that such multidimensional representations are common in cellular decision-making.

  13. Enlightening intracellular complexity of living cells with quantitative phase microscopy

    Science.gov (United States)

    Martinez Torres, C.; Laperrousaz, B.; Berguiga, L.; Boyer Provera, E.; Elezgaray, J.; Nicolini, F. E.; Maguer-Satta, V.; Arneodo, A.; Argoul, F.

    2016-03-01

    The internal distribution of refractive indices (RIs) of a living cell is much more complex than usually admitted in multi-shell models. The reconstruction of RI maps from single phase images has rarely been achieved for several reasons: (i) we still have very little knowledge of the impact of internal macromolecular complexes on the local RI and (ii) phase changes produced by light propagation through the sample are mixed with diffraction effects by internal cell bodies. We propose the implementation a 2D wavelet-based contour chain detection method to distinguish internal boundaries thanks to their greatest optical path difference gradients. These contour chains correspond to the highest image phase contrast and follow the local RI inhomogeneities linked to the intracellular structural intricacy. Their statistics and spatial distribution are morphological indicators for distinguishing cells of different origins and to follow their transformation in pathologic situations. We use this method to compare non adherent blood cells from primary and laboratory culture origins, in healthy and pathological situations (chronic myelogenous leukaemia). In a second part of this presentation, we concentrate on the temporal dynamics of the phase contour chains and we discuss the spectral decomposition of their dynamics in both health and disease.

  14. The Chlamydia psittaci genome: a comparative analysis of intracellular pathogens.

    Directory of Open Access Journals (Sweden)

    Anja Voigt

    Full Text Available Chlamydiaceae are a family of obligate intracellular pathogens causing a wide range of diseases in animals and humans, and facing unique evolutionary constraints not encountered by free-living prokaryotes. To investigate genomic aspects of infection, virulence and host preference we have sequenced Chlamydia psittaci, the pathogenic agent of ornithosis.A comparison of the genome of the avian Chlamydia psittaci isolate 6BC with the genomes of other chlamydial species, C. trachomatis, C. muridarum, C. pneumoniae, C. abortus, C. felis and C. caviae, revealed a high level of sequence conservation and synteny across taxa, with the major exception of the human pathogen C. trachomatis. Important differences manifest in the polymorphic membrane protein family specific for the Chlamydiae and in the highly variable chlamydial plasticity zone. We identified a number of psittaci-specific polymorphic membrane proteins of the G family that may be related to differences in host-range and/or virulence as compared to closely related Chlamydiaceae. We calculated non-synonymous to synonymous substitution rate ratios for pairs of orthologous genes to identify putative targets of adaptive evolution and predicted type III secreted effector proteins.This study is the first detailed analysis of the Chlamydia psittaci genome sequence. It provides insights in the genome architecture of C. psittaci and proposes a number of novel candidate genes mostly of yet unknown function that may be important for pathogen-host interactions.

  15. Erythrocytic ferroportin reduces intracellular iron accumulation, hemolysis, and malaria risk.

    Science.gov (United States)

    Zhang, De-Liang; Wu, Jian; Shah, Binal N; Greutélaers, Katja C; Ghosh, Manik C; Ollivierre, Hayden; Su, Xin-Zhuan; Thuma, Philip E; Bedu-Addo, George; Mockenhaupt, Frank P; Gordeuk, Victor R; Rouault, Tracey A

    2018-03-30

    Malaria parasites invade red blood cells (RBCs), consume copious amounts of hemoglobin, and severely disrupt iron regulation in humans. Anemia often accompanies malaria disease; however, iron supplementation therapy inexplicably exacerbates malarial infections. Here we found that the iron exporter ferroportin (FPN) was highly abundant in RBCs, and iron supplementation suppressed its activity. Conditional deletion of the Fpn gene in erythroid cells resulted in accumulation of excess intracellular iron, cellular damage, hemolysis, and increased fatality in malaria-infected mice. In humans, a prevalent FPN mutation, Q248H (glutamine to histidine at position 248), prevented hepcidin-induced degradation of FPN and protected against severe malaria disease. FPN Q248H appears to have been positively selected in African populations in response to the impact of malaria disease. Thus, FPN protects RBCs against oxidative stress and malaria infection. Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

  16. Microrheometric upconversion-based techniques for intracellular viscosity measurements

    Science.gov (United States)

    Rodríguez-Sevilla, Paloma; Zhang, Yuhai; de Sousa, Nuno; Marqués, Manuel I.; Sanz-Rodríguez, Francisco; Jaque, Daniel; Liu, Xiaogang; Haro-González, Patricia

    2017-08-01

    Rheological parameters (viscosity, creep compliance and elasticity) play an important role in cell function and viability. For this reason different strategies have been developed for their study. In this work, two new microrheometric techniques are presented. Both methods take advantage of the analysis of the polarized emission of an upconverting particle to determine its orientation inside the optical trap. Upconverting particles are optical materials that are able to convert infrared radiation into visible light. Their usefulness has been further boosted by the recent demonstration of their three-dimensional control and tracking by single beam infrared optical traps. In this work it is demonstrated that optical torques are responsible of the stable orientation of the upconverting particle inside the trap. Moreover, numerical calculations and experimental data allowed to use the rotation dynamics of the optically trapped upconverting particle for environmental sensing. In particular, the cytoplasm viscosity could be measured by using the rotation time and thermal fluctuations of an intracellular optically trapped upconverting particle, by means of the two previously mentioned microrheometric techniques.

  17. Characteristic features of intracellular pathogenic Leptospira in infected murine macrophages.

    Science.gov (United States)

    Toma, Claudia; Okura, Nobuhiko; Takayama, Chitoshi; Suzuki, Toshihiko

    2011-11-01

    Leptospira interrogans is a spirochaete responsible for a zoonotic disease known as leptospirosis. Leptospires are able to penetrate the abraded skin and mucous membranes and rapidly disseminate to target organs such as the liver, lungs and kidneys. How this pathogen escape from innate immune cells and spread to target organs remains poorly understood. In this paper, the intracellular trafficking undertaken by non-pathogenic Leptospira biflexa and pathogenic L. interrogans in mouse bone marrow-derived macrophages was compared. The delayed in the clearance of L. interrogans was observed. Furthermore, the acquisition of lysosomal markers by L. interrogans-containing phagosomes lagged behind that of L. biflexa-containing phagosomes, and although bone marrow-derived macrophages could degrade L. biflexa as well as L. interrogans, a population of L. interrogans was able to survive and replicate. Intact leptospires were found within vacuoles at 24 h post infection, suggesting that bacterial replication occurs within a membrane-bound compartment. In contrast, L. biflexa were completely degraded at 24 h post infection. Furthermore, L. interrogans but not L. biflexa, were released to the extracellular milieu. These results suggest that pathogenic leptospires are able to survive, replicate and exit from mouse macrophages, enabling their eventual spread to target organs. © 2011 Blackwell Publishing Ltd.

  18. Collective Dynamics of Intracellular Water in Living Cells

    International Nuclear Information System (INIS)

    Orecchini, A; Sebastiani, F; Paciaroni, A; Petrillo, C; Sacchetti, F; Jasnin, M; Francesco, A De; Zaccai, G; Moulin, M; Haertlein, M

    2012-01-01

    Water dynamics plays a fundamental role for the fulfillment of biological functions in living organisms. Decades of hydrated protein powder studies have revealed the peculiar dynamical properties of hydration water with respect to pure water, due to close coupling interactions with the macromolecule. In such a framework, we have studied coherent collective dynamics in protein and DNA hydration water. State-of-the-art neutron instrumentation has allowed us to observe the propagation of coherent density fluctuations within the hydration shell of the biomolecules. The corresponding dispersion curves resulted to be only slightly affected by the coupling with the macromolecules. Nevertheless, the effects of the interaction appeared as a marked increase of the mode damping factors, which suggested a destructuring of the water hydrogen-bond network. Such results were interpreted as the signature of a 'glassy' dynamical character of macromolecule hydration water, in agreement with indications from measurements of the density of vibrational states. Extending the investigations to living organisms at physiological conditions, we present here an in-vivo study of collective dynamics of intracellular water in Escherichia coli cells. The cells and water were fully deuterated to minimise the incoherent neutron scattering background. The water dynamics observed in the living cells is discussed in terms of the dynamics of pure bulk water and that of hydration water measured in powder samples.

  19. Heme requirement and intracellular trafficking in Trypanosoma cruzi epimastigotes

    International Nuclear Information System (INIS)

    Lara, F.A.; Sant'Anna, C.; Lemos, D.; Laranja, G.A.T.; Coelho, M.G.P.; Reis Salles, I.; Michel, A.; Oliveira, P.L.; Cunha-e-Silva, N.; Salmon, D.; Paes, M.C.

    2007-01-01

    Epimastigotes multiplies in the insect midgut by taking up nutrients present in the blood meal including heme bound to hemoglobin of red blood cell. During blood meal digestion by vector proteases in the posterior midgut, hemoglobin is clipped off into amino acids, peptides, and free heme. In this paper, we compared the heme and hemoglobin uptake kinetics and followed their intracellular trafficking. Addition of heme to culture medium increased epimastigote proliferation in a dose-dependent manner, while medium supplemented with hemoglobin enhanced growth after 3-day lag phase. Medium supplemented with globin-derived peptides stimulated cell proliferation in a dose-independent way. Using Palladium mesoporphyrin IX (Pd-mP) as a fluorescent heme-analog, we observed that heme internalization proceeded much faster than that observed by hemoglobin-rhodamine. Binding experiments showed that parasites accumulated the Pd-mP into the posterior region of the cell whereas hemoglobin-rhodamine stained the anterior region. Finally, using different specific inhibitors of ABC transporters we conclude that a P-glycoprotein homologue transporter is probably involved in heme transport through the plasma membrane

  20. Peripheral neuropathies associated with antibodies directed to intracellular neural antigens.

    Science.gov (United States)

    Antoine, J-C

    2014-10-01

    Antibodies directed to intracellular neural antigens have been mainly described in paraneoplastic peripheral neuropathies and mostly includes anti-Hu and anti-CV2/CRMP5 antibodies. These antibodies occur with different patterns of neuropathy. With anti-Hu antibody, the most frequent manifestation is sensory neuronopathy with frequent autonomic involvement. With anti-CV2/CRMP5 the neuropathy is more frequently sensory and motor with an axonal or mixed demyelinating and axonal electrophysiological pattern. The clinical pattern of these neuropathies is in keeping with the cellular distribution of HuD and CRMP5 in the peripheral nervous system. Although present in high titer, these antibodies are probably not directly responsible for the neuropathy. Pathological and experimental studies indicate that cytotoxic T-cells are probably the main effectors of the immune response. These disorders contrast with those in which antibodies recognize a cell surface antigen and are probably responsible for the disease. The neuronal cell death and axonal degeneration which result from T-cell mediated immunity explains why treating these disorders remains challenging. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  1. Intracellular fate of Ureaplasma parvum entrapped by host cellular autophagy.

    Science.gov (United States)

    Nishiumi, Fumiko; Ogawa, Michinaga; Nakura, Yukiko; Hamada, Yusuke; Nakayama, Masahiro; Mitobe, Jiro; Hiraide, Atsushi; Sakai, Norio; Takeuchi, Makoto; Yoshimori, Tamotsu; Yanagihara, Itaru

    2017-06-01

    Genital mycoplasmas, including Ureaplasma spp., are among the smallest human pathogenic bacteria and are associated with preterm birth. Electron microscopic observation of U. parvum showed that these prokaryotes have a regular, spherical shape with a mean diameter of 146 nm. U. parvum was internalized into HeLa cells by clathrin-mediated endocytosis and survived for at least 14 days around the perinuclear region. Intracellular U. parvum reached endosomes in HeLa cells labeled with EEA1, Rab7, and LAMP-1 within 1 to 3 hr. After 3 hr of infection, U. parvum induced the cytosolic accumulation of galectin-3 and was subsequently entrapped by the autophagy marker LC3. However, when using atg7 -/- MEF cells, autophagy was inadequate for the complete elimination of U. parvum in HeLa cells. U. parvum also colocalized with the recycling endosome marker Rab11. Furthermore, the exosomes purified from infected HeLa cell culture medium included U. parvum. In these purified exosomes ureaplasma lipoprotein multiple banded antigen, host cellular annexin A2, CD9, and CD63 were detected. This research has successfully shown that Ureaplasma spp. utilize the host cellular membrane compartments possibly to evade the host immune system. © 2017 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  2. Prion protein modulates glucose homeostasis by altering intracellular iron.

    Science.gov (United States)

    Ashok, Ajay; Singh, Neena

    2018-04-26

    The prion protein (PrP C ), a mainly neuronal protein, is known to modulate glucose homeostasis in mouse models. We explored the underlying mechanism in mouse models and the human pancreatic β-cell line 1.1B4. We report expression of PrP C on mouse pancreatic β-cells, where it promoted uptake of iron through divalent-metal-transporters. Accordingly, pancreatic iron stores in PrP knockout mice (PrP -/- ) were significantly lower than wild type (PrP +/+ ) controls. Silencing of PrP C in 1.1B4 cells resulted in significant depletion of intracellular (IC) iron, and remarkably, upregulation of glucose transporter GLUT2 and insulin. Iron overloading, on the other hand, resulted in downregulation of GLUT2 and insulin in a PrP C -dependent manner. Similar observations were noted in the brain, liver, and neuroretina of iron overloaded PrP +/+ but not PrP -/- mice, indicating PrP C -mediated modulation of insulin and glucose homeostasis through iron. Peripheral challenge with glucose and insulin revealed blunting of the response in iron-overloaded PrP +/+ relative to PrP -/- mice, suggesting that PrP C -mediated modulation of IC iron influences both secretion and sensitivity of peripheral organs to insulin. These observations have implications for Alzheimer's disease and diabetic retinopathy, known complications of type-2-diabetes associated with brain and ocular iron-dyshomeostasis.

  3. Intracellular Enzymes Contribution to the Biocatalytic Removal of Pharmaceuticals by Trametes hirsuta.

    Science.gov (United States)

    Haroune, Lounès; Saibi, Sabrina; Cabana, Hubert; Bellenger, Jean-Philippe

    2017-01-17

    The use of white rot fungi (WRF) for bioremediation of recalcitrant trace organic contaminants (TrOCs) is becoming greatly popular. Biosorption and lignin modifying enzymes (LMEs) are the most often reported mechanisms of action. Intracellular enzymes, such as cytochrome P450 (CYP450), have also been suggested to contribute. However, direct evidence of TrOCs uptake and intracellular transformation is lacking. The aim of this study was to evaluate the relative contribution of biosorption, extracellular LMEs activity, TrOCs uptake, and intracellular CYP450 on the removal of six nonsteroidal anti-inflammatories (NSAIs) by Trametes hirsuta. Results show that for most tested NSAIs, LMEs activity and biosorption failed to explain the observed removal. Most tested TrOCs are quickly taken up and intracellularly transformed. Fine characterization of intracellular transformation using ketoprofen showed that CYP450 is not the sole intracellular enzyme responsible for intracellular transformation. The contribution of CYP450 in further transformation of ketoprofen byproducts is also reported. These results illustrate that TrOCs transformation by WRF is a more complex process than previously reported. Rapid uptake of TrOCs and intracellular transformation through diverse enzymatic systems appears to be important components of WRF efficiency toward TrOCs.

  4. Determination of Intracellular Vitrification Temperatures for Unicellular Micro Organisms under Conditions Relevant for Cryopreservation.

    Science.gov (United States)

    Fonseca, Fernanda; Meneghel, Julie; Cenard, Stéphanie; Passot, Stéphanie; Morris, G John

    2016-01-01

    During cryopreservation ice nucleation and crystal growth may occur within cells or the intracellular compartment may vitrify. Whilst previous literature describes intracellular vitrification in a qualitative manner, here we measure the intracellular vitrification temperature of bacteria and yeasts under conditions relevant to cryopreservation, including the addition of high levels of permeating and nonpermeating additives and the application of rapid rates of cooling. The effects of growth conditions that are known to modify cellular freezing resistance on the intracellular vitrification temperature are also examined. For bacteria a plot of the activity on thawing against intracellular glass transition of the maximally freeze-concentrated matrix (Tg') shows that cells with the lowest value of intracellular Tg' survive the freezing process better than cells with a higher intracellular Tg'. This paper demonstrates the role of the physical state of the intracellular environment in determining the response of microbial cells to preservation and could be a powerful tool to be manipulated to allow the optimization of methods for the preservation of microorganisms.

  5. Molecular and biochemical characterization of a novel intracellular invertase from Aspergillus niger with transfructosylating activity

    NARCIS (Netherlands)

    Goosen, C.; Yuan, X.L.; Munster, J.M. van; Ram, A.F.J.; Maarel, M.J.E.C. van der; Dijkhuizen, L.

    2007-01-01

    A novel subfamily of putative intracellular invertase enzymes (glycoside hydrolase family 32) has previously been identified in fungal genomes. Here, we report phylogenetic, molecular, and biochemical characteristics of SucB, one of two novel intracellular invertases identified in Aspergillus niger.

  6. Worst-case prediction of normal operating containment temperatures for environmentally qualified equipment

    International Nuclear Information System (INIS)

    Krasnopoler, M.J.; Sundergill, J.E.

    1991-01-01

    Due to issues raised in NRC Information Notice No. 87-65, a southern US nuclear plant was concerned about thermal aging of environmentally qualified (EQ) equipment located in areas of elevated containment temperatures. A method to predict the worst-case monthly temperatures at various zones in the containment and calculate the qualified life using this monthly temperature was developed. Temperatures were predicted for twenty locations inside the containment. Concern about the qualified life of EQ equipment resulted from normal operating temperatures above 120F in several areas of the containment, especially during the summer. At a few locations, the temperature exceeded 140F. Also, NRC Information Notice No. 89-30 reported high containment temperatures at three other nuclear power plants. The predicted temperatures were based on a one-year containment temperature monitoring program. The monitors included permanent temperature monitors required by the Technical Specifications and temporary monitors installed specifically for this program. The temporary monitors were installed near EQ equipment in the expected worst-case areas based on design and operating experience. A semi-empirical model that combined physical and statistical models was developed. The physical model was an overall energy balance for the containment. The statistical model consists of several linear regressions that conservatively relate the monitor temperatures to the bulk containment temperature. The resulting semi-empirical model predicts the worst-case monthly service temperatures at the location of each of the containment temperature monitors. The monthly temperatures are the maximum expected because they are based on the historically worst-case atmospheric data

  7. [Cognitive emotion regulation of patients qualified for implantation of heart rhythm control device].

    Science.gov (United States)

    Ziętalewicz, Urszula; Jędrzejczyk, Jan; Mojkowski, Włodzimierz; Mojkowski, Dariusz

    2016-11-25

    The aim of the artificial heart stimulation is not only saving lives, but also improvement of the quality of life of patients with cardiac arrhythmias. One of the key dimensions of quality of life is psychological functioning. Until now, little research assess this dimension in patients before the implantation of the heart rhythm control device. The aim of the study was to assess the severity of depression and anxiety and the frequency of the used cognitive emotion regulation strategies and to examine the relationship between them. The study group consisted of 60 people qualified for pacemaker implantation (42 PM patients and 18 ICD): 15 women and 45 men ranging in age from 43 to 85. To assess cognitive emotion regulation strategies Cognitive Emotion Regulation Questionnaire was used, and to assess the severity of depression and anxiety - Mood Assessment Questionnaire. Patients with PM more often than patients with ICD use the strategy of Positive Reappraisal (U = 231.50, p = 0.045). There were no statistically significant differences in the frequency of use of other strategies and severity of depression and anxiety. In PM patients there are negative correlations between the severity of depression and anxiety and the use of Acceptance ( τ = -0.380), a Positive Reappraisal ( τ = -0.278), Positive Refocusing ( τ = -0.366) and between the level of anxiety and Putting into Perspective ( τ = -0.402). In ICD patients there was a positive relationship between anxiety and Cathastrophizing ( τ = 0.324). The severity of depression and anxiety, and emotion regulation strategies in patients qualified for PM implantation in comparison with patients qualified for ICD implantation are similar. Both groups of patients show a good adaptation of the psychological.

  8. IE Information Notice No. 85-82: Diesel generator differential protection relay not seismically qualified

    International Nuclear Information System (INIS)

    Jordan, E.L.

    1992-01-01

    Licensees of BWR and PWR nuclear generating stations have reported using a high-speed differential protection relay that has been declared by the manufacturer to be not seismically qualified for the service intended. The relay, GE Model 12CFD, which is being used for emergency diesel generator protection against electrical shorts and grounds at the generator output, has not been seismically qualified to operate in the de-energized state for this application. Generally, the relay operates in the de-energized position before EDG operation. However, it then operates in the energized position whenever the EDG provides an electrical output to essential Class IE components. The safety concern of this issue involves an inadvertent activation of the normally de-energized relay from a seismic event. Momentary activation of the relay, in the circuits examined, blocks the automatic start feature of the EDG during certain emergency plant conditions through operation of another relay in the system. The NRC review of the problems concluded that the relays should be seismically qualified in their energized and de-energized states to preclude their spurious activation during a seismic event. Additionally, review of GE's GER 3069 report, ''Relay Protection of Class IE Systems,'' shows that the 12CFD differential relays were successfully tested for 3.5 g in the energized modes and only 0.75 g in the de-energized mode. Review of GE's GER 3069 report also revealed that GE type IJD differential relays do meet seismic requirements in all modes of operation mentioned above and is the GE recommendation for replacement of the differential relay in question

  9. Intracellular nitrate of marine diatoms as a driver of anaerobic nitrogen cycling in sinking aggregates

    DEFF Research Database (Denmark)

    Kamp, Anja; Stief, Peter; Bristow, Laura A.

    2016-01-01

    % was recovered as nitrite. Hence, aggregate-associated diatoms accumulate nitrate from the surrounding water and sustain complex nitrogen transformations, including loss of fixed nitrogen, in anoxic, pelagic microniches. Additionally, it may be expected that intracellular nitrate not converted before...... store nitrate intracellularly, we explored the fate of intracellular nitrate and its availability for microbial metabolism within anoxic diatom-bacteria aggregates. The ubiquitous nitrate-storing diatom Skeletonema marinoi was studied as both axenic cultures and laboratory-produced diatom......-bacteria aggregates. Stable 15N isotope incubations under dark and anoxic conditions revealed that axenic S. marinoi is able to reduce intracellular nitrate to ammonium that is immediately excreted by the cells. When exposed to a light:dark cycle and oxic conditions, S. marinoi stored nitrate intracellularly...

  10. Data for automated, high-throughput microscopy analysis of intracellular bacterial colonies using spot detection.

    Science.gov (United States)

    Ernstsen, Christina L; Login, Frédéric H; Jensen, Helene H; Nørregaard, Rikke; Møller-Jensen, Jakob; Nejsum, Lene N

    2017-10-01

    Quantification of intracellular bacterial colonies is useful in strategies directed against bacterial attachment, subsequent cellular invasion and intracellular proliferation. An automated, high-throughput microscopy-method was established to quantify the number and size of intracellular bacterial colonies in infected host cells (Detection and quantification of intracellular bacterial colonies by automated, high-throughput microscopy, Ernstsen et al., 2017 [1]). The infected cells were imaged with a 10× objective and number of intracellular bacterial colonies, their size distribution and the number of cell nuclei were automatically quantified using a spot detection-tool. The spot detection-output was exported to Excel, where data analysis was performed. In this article, micrographs and spot detection data are made available to facilitate implementation of the method.

  11. Mycobacterium avium-intracellulare cellulitis occurring with septic arthritis after joint injection: a case report

    Directory of Open Access Journals (Sweden)

    Murdoch David M

    2007-02-01

    Full Text Available Abstract Background Cellulitis caused by Mycobacterium avium-intracellulare has rarely been described. Mycobacterium avium-intracellulare is a rare cause of septic arthritis after intra-articular injection, though the causative role of injection is difficult to ascertain in such cases. Case presentation A 57-year-old with rheumatoid arthritis treated with prednisone and azathioprine developed bilateral painful degenerative shoulder arthritis. After corticosteroid injections into both acromioclavicular joints, he developed bilateral cellulitis centered over the injection sites. Skin biopsy showed non-caseating granulomas, and culture grew Mycobacterium avium-intracellulare. Joint aspiration also revealed Mycobacterium avium-intracellulare infection. Conclusion Although rare, skin and joint infections caused by Mycobacterium avium-intracellulare should be considered in any immunocompromised host, particularly after intra-articular injection. Stains for acid-fast bacilli may be negative in pathologic samples even in the presence of infection; cultures of tissue specimens should always be obtained.

  12. Learning Styles of Pilots Currently Qualified in United States Air Force Aircraft

    Science.gov (United States)

    Kanske, Craig A.

    2001-01-01

    Kolb's Learning Style Inventory was used to identify the predominant learning styles of pilots currently qualified in United States Air Force aircraft. The results indicate that these pilots show a significant preference for facts and things over people and feelings. By understanding the preferred learning styles of the target population, course material can be developed that take advantage of the strengths of these learning styles. This information can be especially useful in the future design of cockpit resource management training. The training program can be developed to demonstrate both that there are different learning styles and that it is possible to take advantage of the relative strengths of each of these learning styles.

  13. [Qualified and emergency specialized surgical care for those with wounds to the extremities].

    Science.gov (United States)

    Iurkevich, V V; Fidarov, E Z; Bauér, V A

    1997-06-01

    Experience of organization of the surgical care in the military hospital to 438 wounded in extremities during armed conflict in Republic of Chechnya is generalized. Maximum reduction of stages of medical evacuation of the wounded in extremities, approaching of the qualified and urgent specialized surgical care directly to the region of battle actions, use of opportunities for it one-moment rendering corresponded to principles of the modern military-medical doctrine. Due to realization of the requirements of the doctrine life of many wounded ++ was saved, terms of treatment, medical and social rehabilitation are reduced. Besides lethality, treatment cost and numbers of transferring to the reserve from the Armed Forces were reduced.

  14. The rhythm and tempo of the game of highly qualified teams

    Directory of Open Access Journals (Sweden)

    V’yacheslav Mulik

    2015-06-01

    Full Text Available Purpose: to set indicators of rhythm and tempo of the game teams of high qualification. Material and Methods: analysis of the scientific-methodical literature, registration of technical-tactical actions, methods of mathematical statistics. The study of competitive activities was conducted with participating teams of world championship 2014. Results: the acticle shows indicators of the rhythm and tempo of the game of well-qualified teams. Conclusions: teams-winners have surpassed teams that concede in terms of indicators of passes the ball, shots at goal, the rhythm of the game, tempo of game.

  15. Les travailleurs statutaires peu qualifiés dans la modernisation des entreprises publiques

    OpenAIRE

    Cultiaux, John; Scius, Marc

    2012-01-01

    Cet article a pour objectif de présenter certains résultats issus de recherches-action menées entre 2001 et 2008 auprès de travailleurs statutaires peu qualifiés et de représentants syndicaux de deux entreprises publiques belges. Endossant un positionnement de cliniciens du travail, nous mettons en évidence les traits essentiels de l’expérience singulière de ces travailleurs mais aussi des circonstances dans lesquelles ils ont fait face aux changements. Observer la modernisation de leur point...

  16. Qualifying Elbow Meters for High Pressure Flow Measurements in an Operating Nuclear Power Plant

    International Nuclear Information System (INIS)

    Chan, A.M.; Maynard, K.J.; Ramundi, J.; Wiklung, E.

    2006-01-01

    To support the installation and use of elbow meters to measure the high pressure emergency coolant injection flow in an operating nuclear station, a test program was performed to qualify: (i) the 'hot' tapping procedure for field application and (ii) the use of elbow meters for accurate flow measurements over the full range of station ECI flow conditions. This paper describes the design conditions and major components of a flow loop used for the elbow meter calibrations. Typical test results are presented and discussed. (authors)

  17. Reporting From the Front Lines: Implementing Oregon's Alternative Payment Methodology in Federally Qualified Health Centers.

    Science.gov (United States)

    Cottrell, Erika K; Hall, Jennifer D; Kautz, Glenn; Angier, Heather; Likumahuwa-Ackman, Sonja; Sisulak, Laura; Keller, Sara; Cameron, David C; DeVoe, Jennifer E; Cohen, Deborah J

    Alternative payment models have been proposed as a way to facilitate patient-centered medical home model implementation, yet little is known about how payment reform translates into changes in care delivery. We conducted site visits, observed operations, and conducted interviews within 3 Federally Qualified Health Center organizations that were part of Oregon's Alternative Payment Methodology demonstration project. Data were analyzed using an immersion-crystallization approach. We identified several care delivery changes during the early stages of implementation, as well as challenges associated with this new model of payment. Future research is needed to further understand the implications of these changes.

  18. Development of viral nanoparticles for efficient intracellular delivery

    Science.gov (United States)

    Wu, Zhuojun; Chen, Kevin; Yildiz, Ibrahim; Dirksen, Anouk; Fischer, Rainer; Dawson, Philip E.; Steinmetz, Nicole F.

    2012-05-01

    Viral nanoparticles (VNPs) based on plant viruses such as Cowpea mosaic virus (CPMV) can be used for a broad range of biomedical applications because they present a robust scaffold that allows functionalization by chemical conjugation and genetic modification, thereby offering an efficient drug delivery platform that can target specific cells and tissues. VNPs such as CPMV show natural affinity to cells; however, cellular uptake is inefficient. Here we show that chemical modification of the CPMV surface with a highly reactive, specific and UV-traceable hydrazone linker allows bioconjugation of polyarginine (R5) cell penetrating peptides (CPPs), which can overcome these limitations. The resulting CPMV-R5 particles were taken up into a human cervical cancer cell line (HeLa) more efficiently than native particles. Uptake efficiency was dependent on the density of R5 peptides on the surface of the VNP; particles displaying 40 R5 peptides per CPMV (denoted as CPMV-R5H) interact strongly with the plasma membrane and are taken up into the cells via an energy-dependent mechanism whereas particles displaying 10 R5 peptides per CPMV (CPMV-R5L) are only slowly taken up. The fate of CPMV-R5 versus native CPMV particles within cells was evaluated in a co-localization time course study. It was indicated that the intracellular localization of CPMV-R5 and CPMV differs; CPMV remains trapped in Lamp-1 positive endolysosomes over long time frames; in contrast, 30-50% of the CPMV-R5 particles transitioned from the endosome into other cellular vesicles or compartments. Our data provide the groundwork for the development of efficient drug delivery formulations based on CPMV-R5.Viral nanoparticles (VNPs) based on plant viruses such as Cowpea mosaic virus (CPMV) can be used for a broad range of biomedical applications because they present a robust scaffold that allows functionalization by chemical conjugation and genetic modification, thereby offering an efficient drug delivery platform

  19. Photonic crystal fiber monitors for intracellular ice formation

    Science.gov (United States)

    Battinelli, Emily; Reimlinger, Mark; Wynne, Rosalind

    2012-04-01

    An all-silica steering wheel photonic crystal fiber (SW-PCF) device with real-time analysis for cellular temperature sensing is presented. Results are provided for water-filled SW-PCF fibers experiencing cooling down near -40°C. Cellular temperature sensors with fast response times are of interest particularly to the study of cryopreservation, which has been influential in applications such as tissue preservation, food quality control, genetic engineering, as well as drug discovery and in- vitro toxin testing. Results of this investigation are relevant to detection of intracellular ice formation (IIF) and better understanding cell freezing at very low temperatures. IIF detection is determined as a function of absorption occurring within the core of the SW-PCF. The SW-PCF has a 3.3μm core diameter, 125μm outer diameter and steering wheel-like air hole pattern with triangular symmetry, with a 20μm radius. One end of a 0.6m length of the SW-PCF is placed between two thermoelectric coolers, filled with ~0.1μL water. This end is butt coupled to a 0.5m length of single mode fiber (SMF), the distal end of the fiber is then inserted into an optical spectrum analyzer. A near-IR light source is guided through the fiber, such that the absorption of the material in the core can be measured. Spectral characteristics demonstrated by the optical absorption of the water sample were present near the 1300-1700nm window region with strongest peaks at 1350, 1410 and 1460nm, further shifting of the absorption peaks is possible at cryogenic temperatures making this device suitable for IIF monitoring applications.

  20. Two-dimensional polyacrylamide gel electrophoresis of intracellular proteins

    International Nuclear Information System (INIS)

    Ojima, N.; Sakamoto, T.; Yamashita, M.

    1996-01-01

    Since two-dimensional electrophoresis was established by O'Farrell for analysis of intracellular proteins of Escherichia coli, it has been applied to separation of proteins of animal cells and tissues, and especially to identification of stress proteins. Using this technique, proteins are separated by isoelectric focusing containing 8 m urea in the first dimension and by SDS-PAGE in the second dimension. The gels are stained with Coomassie Blue R-250 dye, followed by silver staining. In the case of radio-labeled proteins, the gels are dried and then autoradiographed. In order to identify a specific protein separated by two-dimensional electrophoresis, a technique determining the N-terminal amino acid sequence of the protein has been developed recently. After the proteins in the gel were electrotransferred to a polyvinylidene difluoride membrane, the membrane was stained for protein with Commassie Blue and a stained membrane fragment was applied to a protein sequencer. Our recent studies demonstrated that fish cells newly synthesized various proteins in response to heat shock, cold nd osmotic stresses. For example, when cellular proteins extracted from cold-treated rainbow trout cells were subjected to two-dimensional gel electrophoresis, the 70 kDa protein was found to be synthesized during the cold-treatment. N-Terminal sequence analysis showed that the cold-inducible protein was a homolog of mammalian valosin-containing protein and yeast cell division cycle gene product CDC48p. Furthermore, the sequence data were useful for preparing PCR primers and a rabbit antibody against a synthetic peptide to analyze a role for the protein in the function of trout cells and mechanisms for regulation

  1. Intracellular trafficking pathways of Cx43 gap junction channels.

    Science.gov (United States)

    Epifantseva, Irina; Shaw, Robin M

    2018-01-01

    Gap Junction (GJ) channels, including the most common Connexin 43 (Cx43), have fundamental roles in excitable tissues by facilitating rapid transmission of action potentials between adjacent cells. For instance, synchronization during each heartbeat is regulated by these ion channels at the cardiomyocyte cell-cell border. Cx43 protein has a short half-life, and rapid synthesis and timely delivery of those proteins to particular subdomains are crucial for the cellular organization of gap junctions and maintenance of intracellular coupling. Impairment in gap junction trafficking contributes to dangerous complications in diseased hearts such as the arrhythmias of sudden cardiac death. Of recent interest are the protein-protein interactions with the Cx43 carboxy-terminus. These interactions have significant impact on the full length Cx43 lifecycle and also contribute to trafficking of Cx43 as well as possibly other functions. We are learning that many of the known non-canonical roles of Cx43 can be attributed to the recently identified six endogenous Cx43 truncated isoforms which are produced by internal translation. In general, alternative translation is a new leading edge for proteome expansion and therapeutic drug development. This review highlights recent mechanisms identified in the trafficking of gap junction channels, involvement of other proteins contributing to the delivery of channels to the cell-cell border, and understanding of possible roles of the newly discovered alternatively translated isoforms in Cx43 biology. This article is part of a Special Issue entitled: Gap Junction Proteins edited by Jean Claude Herve. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Estrogen receptor of primary breast cancers: evidence for intracellular proteolysis

    International Nuclear Information System (INIS)

    Maaroufi, Younes; Lacroix, Marc; Lespagnard, Laurence; Journé, Fabrice; Larsimont, Denis; Leclercq, Guy

    2000-01-01

    Iodinated oestradiol-labeled oestrogen receptor (ER) isoforms devoid of amino-terminal ABC domains represent about two-thirds of the whole receptor population detected in cytosol samples from human breast cancers. This high frequency could not be ascribed to the expression of truncated mRNAs, or to the proteolysis of the native ER peptide at the time of homogenization or assay, suggesting an intracellular proteolysis. Free amino-terminal and ligand-binding domains maintained together within oligomeric structure(s); increase of ionic strength separated them. The amino-terminal region was consistently detected in the cell nucleus by specific immunohistochemistry leading to the concept of a potential intranuclear association between ER cleavage products and/or other regulatory proteins. We previously reported that about two-thirds of [ 125 I]oestradiol-labelled cytosolic ERs from breast cancer samples eluted as low-molecular-weight isoforms (≤ 37 kDa, size-exclusion fast pressure liquid chromatography [FPLC]). These isoforms failed to adsorb strongly to hydroxylapatite at high ionic strength, a property that was ascribed to receptors devoid of amino-terminal ABC domains. In view of recent data concerning intracellular proteolysis of several transcriptional regulators, the possibility of such behaviour for ER was assessed. The clinical significance of ER measurement in breast cancer cytosols is well established; approximately 50% of ER-positive cases respond to endocrine therapy. Whether such a poor correlation is related to a high proportion of cleaved ER is a question of prime importance. Failure of routine ER assays to discriminate between full-length and cleaved receptors led us to develop an oestradiol-binding assay based on hydroxylapatite adsorption. The aims of the present study were to demonstrate that hydroxylapatite adsorption assay easily identifies cleaved cytosolic ER forms and to assess the origin of such ER forms. Breast cancer cytosols classified as

  3. Intracellular accumulation of potent amiloride analogues by human neutrophils

    International Nuclear Information System (INIS)

    Simchowitz, L.; Woltersdorf, O.W. Jr.; Cragoe, E.J. Jr.

    1987-01-01

    The mechanism of uptake of a series of amiloride derivatives by human neutrophils was investigated using [ 14 C]amiloride and the 14 C-labeled 5-(1-hexahydroazepinyl)-6-bromo analogue (BrMM) which is approximately 500-fold more potent than the parent compound at inhibiting Na+/H+ exchange. At an external concentration of 2 microM, the influx of BrMM at 37 degrees C was rapid, reaching a steady state by approximately 20 min. The rate of BrMM uptake (approximately 25 mumol/liter.min) was approximately 90-fold faster than for the same concentration of amiloride, a finding which correlates with differences in lipid partitioning of the two compounds. Uptake was unrelated to specific binding to Na+/H+ exchange transport sites: influx of either drug was nonsaturable whereas amiloride- and BrMM-mediated inhibition of Na+/H+ countertransport obeyed Michaelis-Menten kinetics with apparent Ki values of approximately 75 and approximately 0.2 microM. Entry occurred exclusively via the neutral (uncharged) forms (pK'a 8.40-8.55). Influx was markedly pH-dependent: it was enhanced by extracellular alkalinization and reduced by acidification. Influx was, however, insensitive to large changes in membrane voltage, thereby implying the protonated (charged) species to be impermeant. About 75% of the total intracellular pool of amiloride, but only approximately 25% of BrMM, is contained within the lysosomes, an expected consequence of the partitioning and subsequent trapping of a weak base within this strongly acidic subcellular compartment. With BrMM, there was a relative approximately 60-fold enrichment in the internal/external water concentration ratio of the drug; the value for amiloride was much less, approximately 4. This disparity is consistent with substantial binding of BrMM to internal constituents, presumably to proteins and/or nucleic acids

  4. Cognitive functions of intracellular mechanisms for contextual amplification.

    Science.gov (United States)

    Phillips, William A

    2017-03-01

    Evidence for the hypothesis that input to the apical tufts of neocortical pyramidal cells plays a central role in cognition by amplifying their responses to feedforward input is reviewed. Apical tufts are electrically remote from the soma, and their inputs come from diverse sources including direct feedback from higher cortical regions, indirect feedback via the thalamus, and long-range lateral connections both within and between cortical regions. This suggests that input to tuft dendrites may amplify the cell's response to basal inputs that they receive via layer 4 and which have synapses closer to the soma. ERP data supporting this inference is noted. Intracellular studies of apical amplification (AA) and of disamplification by inhibitory interneurons targeted only at tufts are reviewed. Cognitive processes that have been related to them by computational, electrophysiological, and psychopathological studies are then outlined. These processes include: figure-ground segregation and Gestalt grouping; contextual disambiguation in perception and sentence comprehension; priming; winner-take-all competition; attention and working memory; setting the level of consciousness; cognitive control; and learning. It is argued that theories in cognitive neuroscience should not assume that all neurons function as integrate-and-fire point processors, but should use the capabilities of cells with distinct sites of integration for driving and modulatory inputs. Potentially 'unifying' theories that depend upon these capabilities are reviewed. It is concluded that evolution of the primitives of AA and disamplification in neocortex may have extended cognitive capabilities beyond those built from the long-established primitives of excitation, inhibition, and disinhibition. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. Radiation-induced adaptive response and intracellular signal transduction pathways

    International Nuclear Information System (INIS)

    Tachibana, Akira

    2009-01-01

    As an essential biological function, cells can sense the radiation even at low dose and respond to it, and which is one of bases of the radiation-induced adaptive response (AR) where effects caused by high dose radiation are reduced by prior exposure to low dose radiation (LDR). Here described are studies of AR in well established m5S cells on the intracellular signal transduction that involves sensing of LDR and transmitting of its signal within the cell network. The first signal for AR yielded by LDR on the cell membrane is exactly unknown though hydrogen peroxide and phorbol ester (PMA) can reportedly cause AR. As PMA activates protein kinase C (PKC) and its inhibitors suppress AR, participation of PKC in AR has been suggested and supported by studies showing PKCα activation by LDR. In addition, p38 mitogen-activated protein kinase (MAPK) is shown to participate in AR by those facts that the enzyme is activated by LDR, a p38 MAPK inhibitor suppresses AR, and PKC inhibitors suppress the enzyme activation, which also suggesting that the signaling from PKC to p38 MAPK can become operative by LDR. However, the possible reverse signaling is also suggested, and thus the activation of positive feedback mechanism is postulated in PKC/p38 MAPK/phospholipase δ1/ PKC pathway. Cells introduced with siRNA against Prkca gene (coding PKCs) produce reduced amount of the enzyme, particularly, of PKCα. In those cells, AR by 5 Gy X-ray is not observed and thereby PKCα is involved in AR. The signaling in AR is only partly elucidated at present as above, and more detailed studies including identification of more PKC subtypes and signaling to DNA repair system are considered necessary. (K.T.)

  6. Intracellular performance of tailored nanoparticle tracers in magnetic particle imaging

    Energy Technology Data Exchange (ETDEWEB)

    Arami, Hamed; Krishnan, Kannan M., E-mail: kannanmk@uw.edu [Department of Materials Science and Engineering, University of Washington, P.O. Box 352120, Seattle, Washington 98195-2120 (United States)

    2014-05-07

    Magnetic Particle Imaging (MPI) is a quantitative mass-sensitive, tracer-based imaging technique, with potential applications in various cellular imaging applications. The spatial resolution of MPI, in the first approximation, improves by decreasing the full width at half maximum (FWHM) of the field-derivative of the magnetization, dm/dH of the nanoparticle (NP) tracers. The FWHM of dm/dH depends critically on NPs’ size, size distribution, and their environment. However, there is limited information on the MPI performance of the NPs after their internalization into cells. In this work, 30 to 150 μg of the iron oxide NPs were incubated in a lysosome-like acidic buffer (0.2 ml, 20 mM citric acid, pH 4.7) and investigated by vibrating sample magnetometry, magnetic particle spectroscopy, transmission electron microscopy, and dynamic light scattering (DLS). The FWHM of the dm/dH curves of the NPs increased with incubation time and buffer to NPs ratio, consistent with a decrease in the median core size of the NPs from ∼20.1 ± 0.98 to ∼18.5 ± 3.15 nm. Further, these smaller degraded NPs formed aggregates that responded to the applied field by hysteretic reversal at higher field values and increased the FWHM. The rate of core size decrease and aggregation were inversely proportional to the concentration of the incubated NPs, due to their slower biodegradation kinetics. The results of this model experiment show that the MPI performance of the NPs in the acidic environments of the intracellular organelles (i.e., lysosomes and endosomes) can be highly dependent on their rate of internalization, residence time, and degradation.

  7. Intracellular magnesium concentrations in dogs with gastric dilatation-volvulus.

    Science.gov (United States)

    Bebchuk, T N; Hauptman, J G; Braselton, W E; Walshaw, R

    2000-11-01

    To quantify and compare intracellular magnesium concentrations (Mgi) in clinically normal dogs (control dogs) and dogs that have gastric dilatation-volvulus (GDV dogs) and to determine whether there is a difference in Mgi and serum magnesium concentrations (Mgs) between GDV dogs with and without cardiac arrhythmias. 41 control dogs and 21 GDV dogs. Rectus abdominis muscle specimens were obtained from control and GDV dogs for determination of Mgi. Blood samples were obtained from GDV dogs for determination of Mgs, and dogs were monitored for 48 hours for cardiac arrhythmias. Muscle specimens were frozen at -40 C, oven dried at 95 C, and digested with concentrated nitric acid. Multielemental analyses were performed by simultaneous/sequential inductively coupled plasma-atomic emission spectroscopy with fixed-cross flow nebulization. The Mg, was standardized to sulfur content to correct for the amount of fat and fascia in the muscle specimen. Mean (+/- SEM) values were recorded in parts per million (ppm). Results-There were no significant differences in Mgi between control (627 +/- 11.1 ppm) and GDV (597 +/- 20.5 ppm) dogs, in Mgi between GDV dogs with (590 +/- 34 ppm) and without (584 +/- 29 ppm) cardiac arrhythmias, and in Mgs between GDV dogs with (1.77 +/- 0.26 ppm) and without (1.51 +/- 0.09 ppm) cardiac arrhythmias. There was no correlation between Mgs and Mgi (R2 = 0.0001). Results indicate that Mg depletion is not pathophysiologically important in dogs with GDV and does not play a role in the cardiac arrhythmias detected in these patients.

  8. IE Information No. 86-104: Unqualified butt splice connectors identified in qualified penetrations

    International Nuclear Information System (INIS)

    Jordan, E.L.

    1992-01-01

    During an NRC equipment qualification (EQ) inspection at Dresden Nuclear Power Station, May 19--23, 1986, a deficiency was discovered concerning a lack of similarity between tested and installed nylon insulated butt splices in EQ qualified GE electrical penetrations. commonwealth Edison sent four sample splices removed from Quad Cities Nuclear Power Station to Wyle Laboratory to further substantiate their qualification for use in a harsh environment. These splices were identical to those installed at Dresden. During the testing performed at Wyle Laboratory December 4--5, 1986, all four samples exhibited excessive leakage currents to ground when exposed to a steam environment. Commonwealth Edison consequently declared the splices unqualified and shut down its Quad Cities Unit 1 to rework the splices by wrapping them with previously qualified tape. Dresden Unit 2 has similarly reworked the splices by wrapping them with tape. Duane Arnold Energy Center also has commenced a shutdown in order to make repairs. The short circuits that occurred appeared to start by condensation entering the splice between the wire insulation and the nylon tubing. The arcing caused insulation degradation that then allowed arcs to pass through the insulation to the enclosure

  9. The lived experience of newly qualified radiographers (1950-1985): An oral history of radiography

    International Nuclear Information System (INIS)

    Decker, Sola

    2009-01-01

    The preparedness for practice of newly qualified healthcare professionals (including radiographers) has been the focus of attention in recent years as the practice environment continues to place great demands on its workforce. This paper reports an aspect of the findings from an oral history project on the career history of radiographers conducted as an academic research investigating the changes that have occurred in radiography and the impact these have had on the profession and the practice of radiography. The main focus of this paper is the lived experience of the informants of the oral history project as newly qualified radiographers. The findings are discussed in the context of current practice environment and developments in radiography. Analysis of the textual materials generated from informants' oral historical accounts, suggest radiographers trained in this time frame perceived their training as 'fit for purpose. The findings further suggest there is a general norm of 'get-on-with-it' as a coping strategy which is still prevalent in the profession today. The paper concludes by drawing on lessons that can be learned from the lived experiences of radiographers' oral historical accounts.

  10. Learning beyond graduation: exploring newly qualified specialists' entrance into daily practice from a learning perspective.

    Science.gov (United States)

    Cuyvers, Katrien; Donche, Vincent; Van den Bossche, Piet

    2016-05-01

    The entrance of newly qualified medical specialists into daily practice is considered to be a stressful period in which curriculum support is absent. Although engaging in both personal and professional learning and development activities is recognized fundamental for lifelong professional competence, research on medical professionals' entrance into practice is scarce. This research aims to contribute to the framework of medical professionals' informal learning and outlines the results of an exploratory study on the nature of learning in daily practice beyond postgraduate training. Eleven newly qualified physicians from different specialized backgrounds participated in a phenomenographic study, using a critical incident method and a grounded theory approach. Results demonstrated that learning in the workplace is, to a large extent, informal and associated with a variety of learning experiences. Analysis shows that experiences related to diagnostics and treatments are important sources for learning. Furthermore, incidents related to communication, changing roles, policy and organization offer learning opportunities, and therefore categorized as learning experiences. A broad range of learning activities are identified in dealing with these learning experiences. More specifically, actively engaging in actions and interactions, especially with colleagues of the same specialty, are the most mentioned. Observing others, consulting written sources, and recognizing uncertainties, are also referred to as learning activities. In the study, interaction, solely or combined with other learning activities, are deemed as very important by specialists in the initial entrance into practice. These insights can be used to develop workplace structures to support the entrance into practice following postgraduate training.

  11. Factors influencing prospective mother with prenatal qualified doctor care among the reproductive women in Bangladesh.

    Science.gov (United States)

    Kiser, Humayun; Nasrin, Tasmina

    2018-12-01

    Maternal and child mortality are the key indicators of health and development of the country. Maternal and child health are interconnected to prenatal care. Consulting a doctor at the prenatal stage will not only ensure mother's and her unborn babies' safety, but also has a great influence to reduce the maternal and infant mortality. In this paper, an attempt has been made to analyze the status of prenatal care provided by the qualified doctor among pregnant mothers in Bangladesh. Data and required information of 8793 reproductive women were collected from the Bangladesh Demographic and Health Survey (BDHS) 2014. Logistic regression model has been used to identify the most significant determinants of the prenatal doctor visits. In this research, it is found that prenatal cares by a qualified doctor during pregnancy depend on several social and demographic characteristics of a woman. It is observed that women staying both urban and rural areas have similar behaviour of caring regarding their pregnancy related complications. Beside this Respondent's age, education, her husband's education and the number of ever born children have significant contribution on prenatal doctor visit. On the other hand, division, religion, husband's desire for children has no effect on it. Overall the model is able to predict 71.65% women into their appropriate group based on these factors.

  12. New technologies applied to radiation protection training: Rp course for technical qualified expert

    International Nuclear Information System (INIS)

    Llorente Herranz, Cristina; Rodriguez, M.; Marco Arboli, Marisa

    2008-01-01

    This paper shows the development and results of the first Course for technical qualified expert in radiation protection (RP) using the advantages of Communication and Information Technologies (CITs). This project in modality b-learning has born as a result of the RP specialised course demand, the necessity of working-private life balance and working-training time combination as well as the geographic widespread of those interested. The methodology selected has been b-learning (blended learning), which consists of a mixture of e-learning plus face-to-face learning. The RP course for technical qualified RP expert has been designed for 10 weeks using online learning methodology and 2 days of face-to-face learning in a radioactive facility to take account practical sessions and the final knowledge evaluation. A multidisciplinary team of experts has elaborated the RP programme and the docent material, following national normative. The material consists of: a) Multimedia material with theoretical content to be visualised online; b) Additional material to be un-load or printed; c) Practical exercises within multimedia content; d) Practical online and face-to-face sessions. (author)

  13. Psychological Properties and Stress Tolerance of High-Qualified Athletes Specializing in Athletic Walking

    Directory of Open Access Journals (Sweden)

    Vasyl Vasylyuk

    2018-04-01

    Full Text Available In the field of sports, there is a strong interest in the behavioral strategies of professional athletes, thus the study of the psychological characteristics of highly skilled athletes is a very topical issue. The article raises the question of the characteristics of the sports environment and the manifestations of stress among athletes. In the paper the psychological features of the personalities of highly qualified athletes who specialize in athletic walking on a scale of neuroticism and extraversion are investigated and the level of neuro-psychological stability of highly skilled athletes specializing in athletic walking is revealed. The main factors of stress in sports activities are: intense competition during a sporting contestation aimed at establishing a record or achieving victory over the rival; maximum physical and psychological stress during the contestation; systematic, long and intense training, which significantly affect the daily regime and everyday life. Two types of stress among athletes are singled out: social-emotional and training. A study of 12 highly qualified athletes who specialize in athletic walking showed that most of them (50 % have ambivert abilities. They are concordants and normostenics that are characterized by significant emotional stability, good adaptability, resistance to external influences. However, among the respondents there were some with low and below average neuropsychic stability.

  14. A space-qualified experiment integrating HTS digital circuits and small cryocoolers

    International Nuclear Information System (INIS)

    Silver, A.; Akerling, G.; Auten, R.

    1996-01-01

    High temperature superconductors (HTS) promise to achieve electrical performance superior to that of conventional electronics. For application in space systems, HTS systems must simultaneously achieve lower power, weight, and volume than conventional electronics, and meet stringent space qualification and reliability requirements. Most effort to date has focused on passive RF/microwave applications. However, incorporation of active microwave components such as amplifiers, mixers, and phase shifters, and on-board high data rate digital signal processing is limited by the power and weight of their spacecraft electronic and support modules. Absence of data on active HTS components will prevent their utilization in space. To validate the feasibility in space of HTS circuits and components based on Josephson junctions, one needs to demonstrate HTS circuits and critical supporting technologies, such as space-qualified packaging and interconnects, closed-cycle cryocooling, and interface electronics. This paper describes the packaging, performance, and space test plan of an integrated, space-qualified experimental package consisting of HTS Josephson junction circuits and all the supporting components for NRL's high temperature superconductor space experiment (HTSSE-II). Most of the technical challenges and approaches are equally applicable to passive and active RF/microwave and digital electronic components, and this experiment will provide valuable validation data

  15. Qualifying commercial grade instruments for use in nuclear power generating stations

    International Nuclear Information System (INIS)

    Lamothe, R.J.; Scally, C.R.

    1983-01-01

    Nuclear environmental qualification of instrumentation has been successfully accomplished by many commercial grade equipment manufacturers. This paper was prepared as a guide to those manufacturers who want some insight into a qualification program. The areas addressed are the regulations and documents, the qualification program, and a case history of a chart recorder qualifications. The principal standards relating to a nuclear qualification program are IEEE Std. 323-1974 IEEE Standard for Qualifying Class 1E Equipment for Nuclear Power Generating Stations, IEEE Std. 344-1975 IEEE Recommended Practices for Seismic Qualification of Class 1E Equipment for Nuclear Power Generating Stations and 10CFR50.49. Previously NUREG 0588 Interim Staff Position on Environmental Qualification of Safety-Related Equipment. These define the intent and purpose of the qualification. The qualification program itself consists of several distinct parts which require explanation, including the determination of qualified life, choice of test samples, selection of appropriate acceptance criteria, aging program, radiation testing, seismic testing, abnormal environment tests and others. The case history illustrates the qualification program and the thought processes involved

  16. A versatile electrical penetration design qualified to IEEE Std. 317-1983

    International Nuclear Information System (INIS)

    Lankenau, W.; Wetherill, T.M.

    1994-01-01

    Although worldwide demand for new construction of nuclear power stations has been on a decline, the available opportunities for the design and construction of qualified electrical penetrations continues to offer challenges, requiring a highly versatile design. Versatility is necessary in order to meet unique customer requirements within the constraints of a design basis qualified to IEEE Std. 317-1983. This paper summarizes such a versatile electrical penetration designed, built and tested to IEEE Std. 317-1983. The principal features are described including major materials of construction. Some of the design constraints such as sealing requirements, and conductor density (including numerical example) are discussed. The requirements for qualification testing of the penetration assembly to IEEE Std. 317-1983 are delineated in a general sense, and some typical test ranges for preconditioning, radiation exposure, and LOCA are provided. The paper concludes by describing ways in which this versatile design has been adapted to meet unique customer requirements in a variety of nuclear power plants

  17. Analysis of the knowledge and opinions of students and qualified dentists regarding the use of computers.

    Science.gov (United States)

    Castelló Castañeda, Coral; Ríos Santos, Jose Vicente; Bullón, Pedro

    2008-01-01

    Dentists are currently required to make multiple diagnoses and treatment decisions every day and the information necessary to achieve this satisfactorily doubles in volume every five years. Knowledge therefore rapidly becomes out of date, so that it is often impossible to remember established information and assimilate new concepts. This may result in a significant lack of knowledge in the future, which would jeopardize the success of treatments. To remedy this situation and to prevent it, we nowadays have access to modern computing systems, with an extensive data base, which helps us to retain the information necessary for daily practice and access it instantaneously. The objectives of this study are therefore to determine how widespread the use of computing is in this environment and to determine the opinion of students and qualified dentists as regards its use in Dentistry. 90 people were chosen to take part in the study, divided into the following groups (students) (newly qualified dentists) (experts). It has been demonstrated that a high percentage (93.30%) use a computer, but that their level of computing knowledge is predominantly moderate. The place where a computer is used most is the home, which suggests that the majority own a computer. Analysis of the results obtained for evaluation of computers in teaching showed that the participants thought that it saved a great deal of time and had great potential for providing an image (in terms of marketing) and they considered it a very innovative and stimulating tool.

  18. Exploring selection and recruitment processes for newly qualified nurses: a sequential-explanatory mixed-method study.

    Science.gov (United States)

    Newton, Paul; Chandler, Val; Morris-Thomson, Trish; Sayer, Jane; Burke, Linda

    2015-01-01

    To map current selection and recruitment processes for newly qualified nurses and to explore the advantages and limitations of current selection and recruitment processes. The need to improve current selection and recruitment practices for newly qualified nurses is highlighted in health policy internationally. A cross-sectional, sequential-explanatory mixed-method design with 4 components: (1) Literature review of selection and recruitment of newly qualified nurses; and (2) Literature review of a public sector professions' selection and recruitment processes; (3) Survey mapping existing selection and recruitment processes for newly qualified nurses; and (4) Qualitative study about recruiters' selection and recruitment processes. Literature searches on the selection and recruitment of newly qualified candidates in teaching and nursing (2005-2013) were conducted. Cross-sectional, mixed-method data were collected from thirty-one (n = 31) individuals in health providers in London who had responsibility for the selection and recruitment of newly qualified nurses using a survey instrument. Of these providers who took part, six (n = 6) purposively selected to be interviewed qualitatively. Issues of supply and demand in the workforce, rather than selection and recruitment tools, predominated in the literature reviews. Examples of tools to measure values, attitudes and skills were found in the nursing literature. The mapping exercise found that providers used many selection and recruitment tools, some providers combined tools to streamline process and assure quality of candidates. Most providers had processes which addressed the issue of quality in the selection and recruitment of newly qualified nurses. The 'assessment centre model', which providers were adopting, allowed for multiple levels of assessment and streamlined recruitment. There is a need to validate the efficacy of the selection tools. © 2014 John Wiley & Sons Ltd.

  19. Directed antigen delivery as a vaccine strategy for an intracellular bacterial pathogen

    Science.gov (United States)

    Bouwer, H. G. Archie; Alberti-Segui, Christine; Montfort, Megan J.; Berkowitz, Nathan D.; Higgins, Darren E.

    2006-03-01

    We have developed a vaccine strategy for generating an attenuated strain of an intracellular bacterial pathogen that, after uptake by professional antigen-presenting cells, does not replicate intracellularly and is readily killed. However, after degradation of the vaccine strain within the phagolysosome, target antigens are released into the cytosol for endogenous processing and presentation for stimulation of CD8+ effector T cells. Applying this strategy to the model intracellular pathogen Listeria monocytogenes, we show that an intracellular replication-deficient vaccine strain is cleared rapidly in normal and immunocompromised animals, yet antigen-specific CD8+ effector T cells are stimulated after immunization. Furthermore, animals immunized with the intracellular replication-deficient vaccine strain are resistant to lethal challenge with a virulent WT strain of L. monocytogenes. These studies suggest a general strategy for developing safe and effective, attenuated intracellular replication-deficient vaccine strains for stimulation of protective immune responses against intracellular bacterial pathogens. CD8+ T cell | replication-deficient | Listeria monocytogenes

  20. Ortholog-based screening and identification of genes related to intracellular survival.

    Science.gov (United States)

    Yang, Xiaowen; Wang, Jiawei; Bing, Guoxia; Bie, Pengfei; De, Yanyan; Lyu, Yanli; Wu, Qingmin

    2018-04-20

    Bioinformatics and comparative genomics analysis methods were used to predict unknown pathogen genes based on homology with identified or functionally clustered genes. In this study, the genes of common pathogens were analyzed to screen and identify genes associated with intracellular survival through sequence similarity, phylogenetic tree analysis and the λ-Red recombination system test method. The total 38,952 protein-coding genes of common pathogens were divided into 19,775 clusters. As demonstrated through a COG analysis, information storage and processing genes might play an important role intracellular survival. Only 19 clusters were present in facultative intracellular pathogens, and not all were present in extracellular pathogens. Construction of a phylogenetic tree selected 18 of these 19 clusters. Comparisons with the DEG database and previous research revealed that seven other clusters are considered essential gene clusters and that seven other clusters are associated with intracellular survival. Moreover, this study confirmed that clusters screened by orthologs with similar function could be replaced with an approved uvrY gene and its orthologs, and the results revealed that the usg gene is associated with intracellular survival. The study improves the current understanding of intracellular pathogens characteristics and allows further exploration of the intracellular survival-related gene modules in these pathogens. Copyright © 2018. Published by Elsevier B.V.

  1. HFE mRNA expression is responsive to intracellular and extracellular iron loading: short communication.

    Science.gov (United States)

    Mehta, Kosha J; Farnaud, Sebastien; Patel, Vinood B

    2017-10-01

    In liver hepatocytes, the HFE gene regulates cellular and systemic iron homeostasis by modulating cellular iron-uptake and producing the iron-hormone hepcidin in response to systemic iron elevation. However, the mechanism of iron-sensing in hepatocytes remain enigmatic. Therefore, to study the effect of iron on HFE and hepcidin (HAMP) expressions under distinct extracellular and intracellular iron-loading, we examined the effect of holotransferrin treatment (1, 2, 5 and 8 g/L for 6 h) on intracellular iron levels, and mRNA expressions of HFE and HAMP in wild-type HepG2 and previously characterized iron-loaded recombinant-TfR1 HepG2 cells. Gene expression was analyzed by real-time PCR and intracellular iron was measured by ferrozine assay. Data showed that in the wild-type cells, where intracellular iron content remained unchanged, HFE expression remained unaltered at low holotransferrin treatments but was upregulated upon 5 g/L (p HFE and HAMP expressions were elevated only at low 1 g/L treatment (p HFE (p HFE mRNA was independently elevated by extracellular and intracellular iron-excess. Thus, it may be involved in sensing both, extracellular and intracellular iron. Repression of HAMP expression under simultaneous intracellular and extracellular iron-loading resembles non-hereditary iron-excess pathologies.

  2. Intracellular Nitrate of Marine Diatoms as a Driver of Anaerobic Nitrogen Cycling in Sinking Aggregates

    Directory of Open Access Journals (Sweden)

    Anja Kamp

    2016-11-01

    Full Text Available Diatom-bacteria aggregates are key for the vertical transport of organic carbon in the ocean. Sinking aggregates also represent pelagic microniches with intensified microbial activity, oxygen depletion in the center, and anaerobic nitrogen cycling. Since some of the aggregate-forming diatom species store nitrate intracellularly, we explored the fate of intracellular nitrate and its availability for microbial metabolism within anoxic diatom-bacteria aggregates. The ubiquitous nitrate-storing diatom Skeletonema marinoi was studied as both axenic cultures and laboratory-produced diatom-bacteria aggregates. Stable 15N isotope incubations under dark and anoxic conditions revealed that axenic S. marinoi is able to reduce intracellular nitrate to ammonium that is immediately excreted by the cells. When exposed to a light:dark cycle and oxic conditions, S. marinoi stored nitrate intracellularly in concentrations > 60 mmol L-1 both as free-living cells and associated to aggregates. Intracellular nitrate concentrations exceeded extracellular concentrations by three orders of magnitude. Intracellular nitrate was used up within 2-3 days after shifting diatom-bacteria aggregates to dark and anoxic conditions. Thirty-one percent of the diatom-derived nitrate was converted to nitrogen gas, indicating that a substantial fraction of the intracellular nitrate pool of S. marinoi becomes available to the aggregate-associated bacterial community. Only 5% of the intracellular nitrate was reduced to ammonium, while 59% was recovered as nitrite. Hence, aggregate-associated diatoms accumulate nitrate from the surrounding water and sustain complex nitrogen transformations, including loss of fixed nitrogen, in anoxic, pelagic microniches. Additionally, it may be expected that intracellular nitrate not converted before the aggregates have settled onto the seafloor could fuel benthic nitrogen transformations.

  3. Dynamics of postirradiation intracellular cysteine and aspartic proteinases profiles in proliferating and nonproliferating mammalian cells

    International Nuclear Information System (INIS)

    Korbelik, M.; Osmak, M.; Suhar, A.; Turk, V.; Skrk, J.

    1990-01-01

    Dynamics of postirradiation intracellular cysteine and aspartic proteinases profiles were examined in proliferating and nonproliferating Chinese hamster fibroblasts (V 79). The results show that there are significant alterations in cysteine and aspartic intracellular proteinases activity already in the early postirradiation period, which are different in proliferating and nonproliferating cells. Irradiation of the cells examined to low doses and up to 15 Gy induced an increase in cysteine proteinases activity in the early postexposure period, while at higher irradiation doses applied, the activity of these proteinases was decreased. These observations suggest that intracellular proteinases are actively participating in process involving recovery from radiation injury or cell killing. (orig.) [de

  4. Mycobacterium intracellulare Pleurisy Identified on Liquid Cultures of the Pleural Fluid and Pleural Biopsy.

    Science.gov (United States)

    Lim, Jong Gu; O, Sei Won; Lee, Ki Dong; Suk, Dong Keun; Jung, Tae Young; Shim, Tae Sun; Chon, Gyu Rak

    2013-03-01

    Pleural effusion is a rare complication in non-tuberculous mycobacterial infection. We report a case of Mycobacterium intracellulare pleuritis with idiopathic pulmonary fibrosis in a 69-year-old man presenting with dyspnea. Pleural effusion revealed lymphocyte dominant exudate. M. intracellulare was identified using a polymerase chain reaction-restriction fragment length polymorphism method and liquid cultures of pleural effusion and pleural biopsy. After combination therapy for M. intracellulare pulmonary disease, the patient was clinically well at a 1-month follow-up.

  5. Concentration-dependent fluorescence live-cell imaging and tracking of intracellular nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Seo, Ji Hye; Joo, Sang-Woo [Department of Chemistry, Soongsil University, Seoul 156-743 (Korea, Republic of); Cho, Keunchang [Logos Biosystems, Incorporated, Anyang 431-070 (Korea, Republic of); Lee, So Yeong, E-mail: leeso@snu.ac.kr, E-mail: sjoo@ssu.ac.kr [Laboratory of Pharmacology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul 151-742 (Korea, Republic of)

    2011-06-10

    Using live-cell imaging techniques we investigated concentration-dependent intracellular movements of fluorescence nanoparticles (NPs) in real-time after their entry into HeLa cells via incubation. Intracellular particle traces appeared to be a mixture of both random and fairly unidirectional movements of the particles. At rather low concentrations of NPs, a majority of the non-random intracellular particle trajectories are assumed to mostly go along microtubule networks after endocytosis, as evidenced from the inhibition test with nocodazole. On the other hand, as the concentrations of NPs increased, random motions were more frequently observed inside the cells.

  6. Concentration-dependent fluorescence live-cell imaging and tracking of intracellular nanoparticles

    International Nuclear Information System (INIS)

    Seo, Ji Hye; Joo, Sang-Woo; Cho, Keunchang; Lee, So Yeong

    2011-01-01

    Using live-cell imaging techniques we investigated concentration-dependent intracellular movements of fluorescence nanoparticles (NPs) in real-time after their entry into HeLa cells via incubation. Intracellular particle traces appeared to be a mixture of both random and fairly unidirectional movements of the particles. At rather low concentrations of NPs, a majority of the non-random intracellular particle trajectories are assumed to mostly go along microtubule networks after endocytosis, as evidenced from the inhibition test with nocodazole. On the other hand, as the concentrations of NPs increased, random motions were more frequently observed inside the cells.

  7. Fluorescent probes and nanoparticles for intracellular sensing of pH values

    Science.gov (United States)

    Shi, Wen; Li, Xiaohua; Ma, Huimin

    2014-12-01

    Intracellular pH regulates a number of cell metabolism processes and its sensing is thus of great importance for cell studies. Among various methods, fluorescent probes have been widely used for sensing intracellular pH values because of their high sensitivity and spatiotemporal resolution capability. In this article, the development of fluorescent probes with good practicability in sensing intracellular pH values and pH variation during 2009 - 2014 is reviewed. These fluorescence probes are divided into two kinds: small molecules and nanoparticles. Photophysical properties, advantages/disadvantages and applications of the two kinds of probes are discussed in detail.

  8. Fluorescent probes and nanoparticles for intracellular sensing of pH values

    International Nuclear Information System (INIS)

    Shi, Wen; Li, Xiaohua; Ma, Huimin

    2014-01-01

    Intracellular pH regulates a number of cell metabolism processes and its sensing is thus of great importance for cell studies. Among various methods, fluorescent probes have been widely used for sensing intracellular pH values because of their high sensitivity and spatiotemporal resolution capability. In this article, the development of fluorescent probes with good practicability in sensing intracellular pH values and pH variation during 2009 − 2014 is reviewed. These fluorescence probes are divided into two kinds: small molecules and nanoparticles. Photophysical properties, advantages/disadvantages and applications of the two kinds of probes are discussed in detail. (topical review)

  9. Intracellular nitrate in sediments of an oxygen-deficient marine basin is linked to pelagic diatoms

    DEFF Research Database (Denmark)

    Kamp, Anja; Petro, Caitlin; Røy, Hans

    2018-01-01

    Intracellular nitrate is an important electron acceptor in oxygen-deficient aquatic environments, either for the nitrate-storing microbes themselves, or for ambient microbial communities through nitrate leakage. This study links the spatial distribution of intracellular nitrate with the abundance...... and identity of nitrate-storing microbes in sediments of the Bornholm Basin, an environmental showcase for severe hypoxia. Intracellular nitrate (up to 270 nmol cm−3 sediment) was detected at all 18 stations along a 35-km transect through the basin and typically extended as deep as 1.6 cm into the sediment...

  10. Engineering intracellular active transport systems as in vivo biomolecular tools.

    Energy Technology Data Exchange (ETDEWEB)

    Bachand, George David; Carroll-Portillo, Amanda

    2006-11-01

    applications. Further development could potentially enable selective capture of intracellular antigens, targeted delivery of therapeutic agents, or disruption of the transport systems and consequently the infection and pathogenesis cycle of biothreat agents.

  11. Rickettsia phylogenomics: unwinding the intricacies of obligate intracellular life.

    Directory of Open Access Journals (Sweden)

    Joseph J Gillespie

    Full Text Available BACKGROUND: Completed genome sequences are rapidly increasing for Rickettsia, obligate intracellular alpha-proteobacteria responsible for various human diseases, including epidemic typhus and Rocky Mountain spotted fever. In light of phylogeny, the establishment of orthologous groups (OGs of open reading frames (ORFs will distinguish the core rickettsial genes and other group specific genes (class 1 OGs or C1OGs from those distributed indiscriminately throughout the rickettsial tree (class 2 OG or C2OGs. METHODOLOGY/PRINCIPAL FINDINGS: We present 1823 representative (no gene duplications and 259 non-representative (at least one gene duplication rickettsial OGs. While the highly reductive (approximately 1.2 MB Rickettsia genomes range in predicted ORFs from 872 to 1512, a core of 752 OGs was identified, depicting the essential Rickettsia genes. Unsurprisingly, this core lacks many metabolic genes, reflecting the dependence on host resources for growth and survival. Additionally, we bolster our recent reclassification of Rickettsia by identifying OGs that define the AG (ancestral group, TG (typhus group, TRG (transitional group, and SFG (spotted fever group rickettsiae. OGs for insect-associated species, tick-associated species and species that harbor plasmids were also predicted. Through superimposition of all OGs over robust phylogeny estimation, we discern between C1OGs and C2OGs, the latter depicting genes either decaying from the conserved C1OGs or acquired laterally. Finally, scrutiny of non-representative OGs revealed high levels of split genes versus gene duplications, with both phenomena confounding gene orthology assignment. Interestingly, non-representative OGs, as well as OGs comprised of several gene families typically involved in microbial pathogenicity and/or the acquisition of virulence factors, fall predominantly within C2OG distributions. CONCLUSION/SIGNIFICANCE: Collectively, we determined the relative conservation and

  12. pH-sensitive intracellular photoluminescence of carbon nanotube-fluorescein conjugates in human ovarian cancer cells

    International Nuclear Information System (INIS)

    Chen, M T; Ishikawa, F N; Gundersen, M A; Gomez, L M; Vernier, P T; Zhou, C

    2009-01-01

    To add to the understanding of the properties of functionalized carbon nanotubes in biological applications, we report a monotonic pH sensitivity of the intracellular fluorescence emission of single-walled carbon nanotube-fluorescein carbazide (SWCNT-FC) conjugates in human ovarian cancer cells. Light-stimulated intracellular hydrolysis of the amide linkage and localized intracellular pH changes are proposed as mechanisms. SWCNT-FC conjugates may serve as intracellular pH sensors.

  13. DMPD: Ubiquitin: tool and target for intracellular NF-kappaB inhibitors. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 16982211 Ubiquitin: tool and target for intracellular NF-kappaB inhibitors. Wullaer...vg) (.html) (.csml) Show Ubiquitin: tool and target for intracellular NF-kappaB inhibitors. PubmedID 1698221...1 Title Ubiquitin: tool and target for intracellular NF-kappaB inhibitors. Author

  14. Vitamin D is positively associated with sperm motility and increases intracellular calcium in human spermatozoa

    DEFF Research Database (Denmark)

    Blomberg Jensen, Martin; Bjerrum, Poul J; Jessen, Torben E

    2011-01-01

    BACKGROUND The vitamin D receptor (VDR) is expressed in human spermatozoa, and VDR-knockout mice and vitamin D (VD) deficiency in rodents results in impaired fertility, low sperm counts and a low number of motile spermatozoa. We investigated the role of activated VD (1,25(OH)(2)D(3)) in human...... spermatozoa and whether VD serum levels are associated with semen quality. METHODS Cross-sectional association study of semen quality and VD serum level in 300 men from the general population, and in vitro studies on spermatozoa from 40 men to investigate the effects of VD on intracellular calcium, sperm......M). 1,25(OH)(2)D(3) increased intracellular calcium concentration in human spermatozoa through VDR-mediated calcium release from an intracellular calcium storage, increased sperm motility and induced the acrosome reaction in vitro. CONCLUSIONS 1,25(OH)(2)D(3) increased intracellular calcium...

  15. Functionalized linear poly(amidoamine)s are efficient vectors for intracellular protein delivery

    NARCIS (Netherlands)

    Coué, G.M.J.P.C.; Engbersen, Johannes F.J.

    2011-01-01

    An effective intracellular protein delivery system was developed based on functionalized linear poly(amidoamine)s (PAAs) that form self-assembled cationic nanocomplexes with oppositely charged proteins. Three differently functionalized PAAs were synthesized, two of these having repetitive disulfide

  16. Diffusive spatio-temporal noise in a first-passage time model for intracellular calcium release

    KAUST Repository

    Flegg, Mark B.; Rüdiger, Sten; Erban, Radek

    2013-01-01

    The intracellular release of calcium from the endoplasmic reticulum is controlled by ion channels. The resulting calcium signals exhibit a rich spatio-temporal signature, which originates at least partly from microscopic fluctuations. While

  17. Sphingosine-1-phosphate receptors: Zooming in on ligand-induced intracellular trafficking and its functional implications

    NARCIS (Netherlands)

    Verzijl, Dennis; Peters, Stephan L. M.; Alewijnse, Astrid E.

    2010-01-01

    Regulatory processes including receptor phosphorylation and intracellular trafficking, also referred to as receptor internalization, are important processes to terminate G protein-coupled receptor (GPCR) signaling. Compelling evidence now indicates that internalization of a receptor is not

  18. Conditions With High Intracellular Glucose Inhibit Sensing Through Glucose Sensor Snf3 in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Karhumaa, Kaisa; Wu, B.Q.; Kielland-Brandt, Morten

    2010-01-01

    as for amino acids. An alternating-access model of the function of transporter-like sensors has been previously suggested based on amino acid sensing, where intracellular ligand inhibits binding of extracellular ligand. Here we studied the effect of intracellular glucose on sensing of extracellular glucose...... through the transporter-like sensor Snf3 in yeast. Sensing through Snf3 was determined by measuring degradation of Mth1 protein. High intracellular glucose concentrations were achieved by using yeast strains lacking monohexose transporters which were grown on maltose. The apparent affinity...... of extracellular glucose to Snf3 was measured for cells grown in non-fermentative medium or on maltose. The apparent affinity for glucose was lowest when the intracellular glucose concentration was high. The results conform to an alternating-access model for transporter-like sensors. J. Cell. Biochem. 110: 920...

  19. Intracellular NAD(H) levels control motility and invasion of glioma cells.

    NARCIS (Netherlands)

    Horssen, R. van; Willemse, M.P.; Haeger, A.; Attanasio, F.; Guneri, T.; Schwab, A.; Stock, C.M.; Buccione, R.; Fransen, J.A.M.; Wieringa, B.

    2013-01-01

    Oncogenic transformation involves reprogramming of cell metabolism, whereby steady-state levels of intracellular NAD(+) and NADH can undergo dramatic changes while ATP concentration is generally well maintained. Altered expression of nicotinamide phosphoribosyltransferase (NAMPT), the rate-limiting

  20. Multi locus sequence typing of Chlamydiales: clonal groupings within the obligate intracellular bacteria Chlamydia trachomatis

    NARCIS (Netherlands)

    Pannekoek, Yvonne; Morelli, Giovanna; Kusecek, Barica; Morré, Servaas A.; Ossewaarde, Jacobus M.; Langerak, Ankie A.; van der Ende, Arie

    2008-01-01

    BACKGROUND: The obligate intracellular growing bacterium Chlamydia trachomatis causes diseases like trachoma, urogenital infection and lymphogranuloma venereum with severe morbidity. Several serovars and genotypes have been identified, but these could not be linked to clinical disease or outcome.