Science.gov

Sample records for igi nyats rk

  1. Legacy of the IGY

    SciTech Connect

    Friedman, H.

    1983-08-09

    The success of the International Geophysical Year (IGY), which ended in 1958, has prompted contemporary geoscientists to consider the possibility of a second-generation IGY, tentatively designated the International Geosphere-Biosphere Program (IGBP). Unlike the IGY, which was planned to run for less than two years, the IGBP must cover one or two decades due to the many geosphere-biosphere cycles which encompass periods on at least that scale. Attention is given to interdisciplinary coincidence of interest among the fields of research covered.

  2. My IGY in Antarctica

    NASA Technical Reports Server (NTRS)

    Bentley, Charles

    2012-01-01

    Dr Charles Bentley is the A.P. Crary Professor Emeritus of Geophysics, Department of Geology and Geophysics, University of Wisconsin-Madison. Dr. Bentley joined the Arctic Institute of North America in 1956 to participate in International Geophysical Year (IGY)-related activities in the Antarctic. He wintered over consecutively in 1957 and 1958 at Byrd Station, a station in the interior of West Antarctica that housed 24 men each winter - 12 Navy support people and 12 civilian scientists/technicians. During the austral summers, he also participated in over-snow traverses, first as co-leader, then leader (the other coleader went home after the first year). These traverses consisted of six men and three vehicles, and lasted several months. These traverses covered more than 1609 kilometers (1000 miles) of largely unmapped and unphotographed terrain. During these traverses, connections to Byrd Station were by radio (daily, when the transmission conditions were good enough) and roughly every 2 weeks by resupply flight.

  3. An ELISA for quantifying quail IgY and characterizing maternal IgY transfer to egg yolk in several quail strains.

    PubMed

    Murai, Atsushi; Kakiuchi, Misako; Hamano, Takahito; Kobayashi, Misato; Tsudzuki, Masaoki; Nakano, Mikiharu; Matsuda, Yoichi; Horio, Fumihiko

    2016-07-01

    In avian species, maternal blood immunoglobulin Y (IgY) is transferred to the egg yolks of maturing oocytes, but the mechanism underlying this transfer is unknown. To gain insight into the mechanism of maternal IgY transfer in quail, we established an enzyme-linked immunosorbent assay (ELISA) for the quantitation of quail IgY. We characterized strain differences in blood and egg yolk IgY concentrations and exogenously injected IgY-Fc uptakes into egg yolks. A specific rabbit polyclonal antibody to quail IgY was raised for the ELISA. Blood and egg yolk IgY concentrations were determined in six quail strains (one inbred strain, L; four closed population strains, AWE, DB, PS, WE; one commercial strain, Commercial). The birds were also injected with digoxigenin-labeled quail IgY-Fc, and its uptakes into laid eggs were compared. The strain difference in blood and egg yolk IgY concentrations was at most 2.5-fold, between PS and AWE. The rank order of IgY concentrations was AWE, Commercial, DB, L≥WE≥PS. A significant positive correlation (|R|=0.786) between individual blood IgY and egg yolk IgY and the concentrated egg yolk IgY (1.5-2-fold) against blood IgY was observed. Interestingly, there was a significant inverse correlation (|R|=0.452) between injected IgY-Fc uptakes and the blood IgY concentration, implying competition of the injected IgY-Fc and blood IgY in the process of IgY uptake into egg yolks. In conclusion, we successfully determined blood and egg yolk IgY concentrations in various quail strains by a quail IgY-specific ELISA. The concentrated egg yolk IgY against the blood IgY and the inverse relationship of exogenous IgY-Fc uptake against the blood IgY supports the existence of a selective IgY transport mechanism in avian maturing oocytes. PMID:27269788

  4. Chronobiological studies of chicken IgY: monitoring of infradian, circadian and ultradian rhythms of IgY in blood and yolk of chickens.

    PubMed

    He, Jin-Xin; Thirumalai, Diraviyam; Schade, Rüdiger; Zhang, Xiao-Ying

    2014-08-15

    IgY is the functional equivalent of mammalian IgG found in birds, reptiles and amphibians. Many of its biological aspects have been explored with different approaches. In order to evaluate the rhythmicity of serum and yolk IgY, four chickens were examined and reared under the same conditions. To monitor biological oscillations of IgY in yolk and serum, the eggs and blood samples were collected over a 60 day period and the rhythm of yolk and serum IgY was determined by direct-ELISA. Results indicated that, there is a significant circaseptan rhythm in yolk IgY and circaquattran rhythm in serum IgY. The serum IgY concentration reached a peak in the morning, decreased to a minimum during the daytime and increased again at night revealing a significant circadian rhythm was superimposed by an ultradian rhythm. These data are suited to address the controversies concerning the IgY concentration in egg yolk and blood of laying hens. In addition, this study raised new questions, if the different rhythms in yolk and serum are concerned. PMID:24998020

  5. The IGY of 1957-58: Its Place in Science, Politics, and History

    NASA Astrophysics Data System (ADS)

    Needell, A.

    2006-12-01

    The International Geophysical Year (IGY) took place during a momentous period of change in the relationship between science, society and its governing institutions. Its origins, structure, goals, and execution reflect the powerful social, cultural and ideological forces that were shaping national and international politics of the period. The IGY also contributed in important ways to the evolution science-government relations throughout the world. This presentation will focus on the social and political contexts within which the IGY was planned and carried in the United States, exploring the significance for the IGY of contemporary academic exchanges over the social aspects of science during the tumultuous 1950s and 60s.

  6. Generation and application of anti-ouabain IgY antibodies.

    PubMed

    Zhang, Ming Juan; Yang, Jun; Zhu, Can Zhan; Duan, Zong Ming; Niu, Xiao Lin

    2011-12-01

    Ouabain is a bioactive hapten and is very difficult to be accurately quantified because of the lack of useful reagents. Furthermore, where ouabain is produced in the adrenal glands has not been identified. In this study, ouabain-BSA was generated for immunizing the laying hens to generate ouabain-specific IgY antibodies in chicken eggs. The anti-ouabain IgY antibodies were detected in eggs 1 week after the last immunization and their concentrations increased with time. The highest concentrations of anti-ouabain IgY antibodies reached at 1:10,240 for ELISA 5 weeks after immunization and maintained for 4 weeks in chicken eggs. Following PEG precipitation, an average of 8.5 mg of anti-ouabain IgY antibodies with a purity of 87.6% was achieved from a single egg. Further analysis revealed that the anti-ouabain IgY antibodies had little immunoreactivity to hydrocortisone, dexamethasone, cedilanid, and digoxin, indicating their high specificity, and the purified IgY antibodies effectively detected endogenous ouabain in the cytoplasm of cells predominately in the zona reticularis of rat and human adrenal glands, indicating their high immunoreactivity. Given that IgY has an unique structure and bioactive features, the generated anti-ouabain IgY antibodies may be used as a new reagent for accurately quantifying ouabain in biological studies. PMID:21785972

  7. Fluorescence polarization immunoassay using IgY antibodies for detection of valnemulin in swine tissue

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Immunoglobulin Y (IgY) is derived from egg yolk and has been identified as a cheap and high-yield immunoreagent. The application of IgY in immunoassay for the detection of chemical contaminants in food samples has rarely been reported. In this work, we describe a rapid and sensitive fluorescence p...

  8. The in vitro and in vivo efficacy of hen IgY against Vibrio parahaemolyticus and Vibrio vulnificus.

    PubMed

    Kassim, Neema; Mtenga, Adelard B; Shim, Won-Bo; Chung, Duck-Hwa

    2012-10-01

    The inhibitory effect of IgY against Vibrio parahaemolyticus and Vibrio vulnificus responsible for seafood-borne diseases was investigated in this study. Water-soluble fractions (WSF) of protein containing IgYs were isolated from the egg yolk of hens initially immunized with formalin inactivated V. parahaemolyticus or V. vulnificus. Protein, total and specific IgY contents of the WSF were determined. The inhibitory and protective effects of IgYs on the growth of V. parahaemolyticus and V. vulnificus were assayed in liquid medium and in mice. IgYs showed high affinity to their corresponding antigens with high titer from day 28 onwards. Protein contents and total IgY concentrations remained stable throughout the immunization period, whereas specific IgY concentrations increased steadily and reached a plateau at day 49. Specific IgY powder (150 mg/ml) significantly inhibited further multiplication of both V. parahaemolyticus and V. vulnificus in liquid medium as compared with the control IgY. The bacteria count in mice feces was lower in mice pretreated with specific IgYs than in those pretreated with PBS or control IgY. Higher survival of mice was observed in the experimental groups pretreated with either anti-V. parahaemolyticus (75% survival) or anti-V. vulnificus (87% survival) IgYs, compared with those in the control groups pretreated with PBS or nonspecific IgY. All mice in the control groups died within three days after bacteria inoculation; hence, the protective effect of specific IgYs against infection caused by V. parahaemolyticus and V. vulnificus was demonstrated. PMID:23075795

  9. Some Legacies from the IGY to the IHY

    NASA Astrophysics Data System (ADS)

    Helliwell, R.

    One thing we learned from the IGY-sponsored Whistlers-West program was to urge operators of field stations to watch for unexpected events in the incoming data. Several such serendipitous discoveries of fundamental importance to magnetospheric physics were found during (as well as after) the IGY period 1957-58. One example is the Trimpi effect, which is the perturbation of the amplitude and the related phase of the signal from a vlf transmitter (eg. NAA, NSS, NPG) caused by a lightning whistler that scatters energetic electrons out of the radiation belt into the ionosphere. These precipitated electrons create a temporary patch of ionization that alters the local conductivity of the E region and causes scattering of the subionosphere vlf signal. Another example is the unexpected triggering of chorus-like emissions by the dashes of morse code signals from communication stations of the U.S. Navy [e.g. NPG, NSS, NAA]). This finding led to the development of a vlf (1-30 kHz) wave injection facility at Siple Station, Antarctica. This facility was operated for 15 years, using two broadband transmitters in sequence. The latter and more flexible transmitter could transmit simultaneously on two different carrier frequencies, each of which could be modulated in both amplitude and frequency. The results of these controlled experiments greatly increased our knowledge of many non-linear WPI effects that collectively have been called the Coherent Wave Instability (CWI), as reviewed in Helliwell, R.A., A tale of two magnetospheres, J. Atmos. Solar Terr. Physics, 1107- 1116, 2001. An example of a possible future study for the IHY is a search for whistler-mode (WM) signals within a CME (coronal mass ejection). It is predicated on the well- known observation of high intensity spontaneous chorus in the equatorial plane of the magnetosphere beyond the plasmapause, where no triggering signals from the ground are seen; suggesting that a CME may constitute a somewhat similar environment

  10. DNA-designed avian IgY antibodies: novel tools for research, diagnostics and therapy.

    PubMed

    Cova, Lucyna

    2005-12-01

    We have recently demonstrated, using the duck Hepatitis B virus (DHBV) model, closely related to human HBV, that following DNA immunization of breeding ducks with a plasmid encoding the targeted protein, specific and biologically active IgY (egg yolk immunoglobulines) are vertically transmitted from their serum into the egg yolk from which they can be extracted and purified. Thus an egg can be considered as a small "factory" for antibody production, since about 60-100 mg of purified IgY can be obtained from each egg yolk of a DNA-immunized duck. One of the major advantages of this new method of "DNA-designed" IgY antibodies is their production via immunization with a gene vector that expresses a corresponding antibody in situ in the cells of an avian host. Therefore this approach allows direct generation of antibodies from plasmid DNA and avoids the costly and tedious preparation of purified antigens required for conventional antibody production. In addition, duck IgY are of remarkable high affinity, avidity and are highly neutralizing. Moreover, the epitope pattern of IgY generated by DNA immunization of ducks is closely related to that observed in viral infection. Such duck IgY are also of particular value as immunodiagnostic tools, since they do not cross-react serologically with mammalian immunoglobulins and complement. Because IgY are resistant to the gastric barrier, the recently described DNA-designed IgY specific to H. pylori Urease B can be of particular interest for passive immunotherapy of gastrointestinal tract infections. Another interesting application is the recent generation in our laboratory of DNA-designed IgY antibodies specific to HBsAg mutants. These antibodies are currently being used to design new diagnostic assay for detection of HBV mutants that are undetectable by actual tests. Moreover, this approach allowing a quick and inexpensive production of a new generation of antibodies will provide pertinent tools to link the fields of genomics

  11. Egg yolk IgY antibodies: A therapeutic intervention against group A rotavirus in calves.

    PubMed

    Vega, C; Bok, M; Saif, L; Fernandez, F; Parreño, V

    2015-12-01

    Bovine group A rotavirus (RVA) is considered the major cause of diarrhea in intensively reared neonatal calves. Chicken egg yolk antibodies (IgY) are efficient in protecting neonatal calves from RVA diarrhea; however, the value of this intervention in calves once diarrhea has appeared is unclear. The aim of the present study was to evaluate the application of RVA-specific IgY as a passive treatment in those cases. The experimental groups were: G1=RVA-specific IgY treatment; G2=no Ab treatment; and G3=colostrum deprived+no Ab treatment. IgY treatment significantly reduced virus shedding, diarrhea duration and severity compared to G2 and G3 calves. However, it caused a partial suppression of systemic Ab responses to RVA that could be associated with less severe diarrhea. The oral treatment with IgY for 7days was associated with significantly higher antibody secreting cell responses in the calves compared with other groups of animals. PMID:26679788

  12. Efficacy of specific egg yolk immunoglobulin (IgY) to bovine mastitis caused by Staphylococcus aureus.

    PubMed

    Zhen, Yu-Hong; Jin, Li-Ji; Li, Xiao-Yu; Guo, Jie; Li, Zhi; Zhang, Bao-Jing; Fang, Rui; Xu, Yong-Ping

    2009-02-01

    The objective of this study was to estimate the efficacy of specific egg yolk immunoglobulin (IgY) to bovine mastitis caused by Staphylococcus aureus. Eighteen lactating cows with clinical mastitis and 18 lactating cows with experimental mastitis (1 quarter per cow) were randomly assigned to three treatments: IgY (20mg/ml) infusion, penicillin (100mg/ml) infusion and no infusion. Treatments for clinical mastitis and experimental mastitis were performed by a 6-day course of intramammary infusion with a dosage of 10ml at an interval of 12h. Milk samples were collected at morning milking time for testing color, clot, somatic cell counts (SCC) and bacterial count. For most of the cows treated with IgY and penicillin, the milk color and clot recovered to normal form during the therapy course. The milk SCCs and bacterial counts of treated cows decreased compared to those of untreated cows (p<0.05). The cure rates by IgY for experimental and clinical mastitis were 83.3% and 50%, respectively, and those by penicillin were 66.7% and 33.3%, respectively. These results showed the potential of specific IgY to be an alternative therapy for mastitis caused by S. aureus. PMID:18774241

  13. Fluorescence polarization immunoassay using IgY antibodies for detection of valnemulin in swine tissue.

    PubMed

    Zhang, Huiyan; Mi, Tiejun; Khan, Oleg Yu; Sheng, Yajie; Eremin, Sergei A; Beier, Ross C; Zhang, Suxia; Shen, Jianzhong; Wang, Zhanhui

    2015-10-01

    Immunoglobulin Y (IgY) is derived from egg yolk and has been identified as a cheap and high-yield immunoreagent. The application of IgY in immunoassays for the detection of chemical contaminants in food samples has rarely been reported. In this work, we describe a rapid and sensitive fluorescence polarization immunoassay (FPIA) for valnemulin (VAL) using IgY which was produced using a previously prepared immunogen. Three fluorophore-labeled VAL tracers were synthesized and the sensitivity of the best tracer (VAL-DTAF) in the optimized FPIA with antibody IgY100 demonstrated an IC50 value of 12 ng mL(-1) in buffer. After evaluation of several extraction procedures, acidified acetonitrile was selected to extract VAL from swine tissue. The recoveries of VAL in spiked swine tissue at three levels (50, 100, and 200 μg kg(-1)) were higher than 79% with coefficients of variation (CVs) lower than 12%. The limit of detection (LOD) of the FPIA in swine tissue was 26 μg kg(-1) and was lower than the maximum residue limit (MRL) of VAL set by the European Union. The study showed that IgY could be a good substitute for IgG when developing a high-throughput assay for chemical residues. PMID:26277187

  14. A comparative evaluation of six principal IgY antibody extraction methods.

    PubMed

    Ren, Hao; Yang, Wenjing; Thirumalai, Diraviyam; Zhang, Xiaoying; Schade, Rüdiger

    2016-03-01

    Egg yolk has been considered a promising source of antibodies. Our study was designed to compare six principal IgY extraction methods (water dilution, polyethylene glycol [PEG] precipitation, caprylic acid extraction, chloroform extraction, phenol extraction, and carrageenan extraction), and to assess their relative extraction efficiencies and the purity of the resulting antibodies. The results showed that the organic solvents (chloroform or phenol) minimised the lipid ratio in the egg yolk. The water dilution, PEG precipitation and caprylic acid extraction methods resulted in high yields, and antibodies purified with PEG and carrageenan exhibited high purity. Our results indicate that phenol extraction would be more suitable for preparing high concentrations of IgY for non-therapeutic usage, while the water dilution and carrageenan extraction methods would be more appropriate for use in the preparation of IgY for oral administration. PMID:27031600

  15. IGI (the Italian Grid initiative) and its impact on the Astrophysics community

    NASA Astrophysics Data System (ADS)

    Pasian, F.; Vuerli, C.; Taffoni, G.

    IGI - the Association for the Italian Grid Infrastructure - has been established as a consortium of 14 different national institutions to provide long term sustainability to the Italian Grid. Its formal predecessor, the Grid.it project, has come to a close in 2006; to extend the benefits of this project, IGI has taken over and acts as the national coordinator for the different sectors of the Italian e-Infrastructure present in EGEE. IGI plans to support activities in a vast range of scientificdisciplines - e.g. Physics, Astrophysics, Biology, Health, Chemistry, Geophysics, Economy, Finance - and any possible extensions to other sectors such as Civil Protection, e-Learning, dissemination in Universities and secondary schools. Among these, the Astrophysics community is active as a user, by porting applications of various kinds, but also as a resource provider in terms of computing power and storage, and as middleware developer.

  16. Eliciting antigen-specific egg-yolk IgY with naked DNA.

    PubMed

    Romito, M; Viljoen, G J; Du Plessis, D H

    2001-09-01

    Immunization with naked DNA was used to elicit chicken egg yolk antibodies (IgY). Layer hens were inoculated with plasmid DNA encoding the enhanced green fluorescent protein, the fusion protein of Newcastle disease virus, and VP2 of African horse sickness virus. IgY was extracted from egg yolks by polyethylene glycol precipitation. Specific antibodies were present in the yolks of eggs from hens immunized with each of the three different plasmids. This approach to raising polyclonal antibodies obviates the need to produce and purify large quantities of proteins for immunization and can potentially yield large amounts of diagnostically or therapeutically useful reagents. PMID:11570510

  17. Pre-IGY Ionosphere Over Washington D.C

    NASA Astrophysics Data System (ADS)

    Rice, D. D.; Sojka, J. J.; Eccles, J. V.; Hunsucker, R. D.

    2012-12-01

    A data recovery study has been sponsored by the NSF to determine how successfully the ionosphere during a pre-IGY era can be inferred from archived ionogram films. This pilot study targets the Washington, DC ionosonde WA938 located at Ft. Belvoir in Fairfax, VA (38.7° N, -77.1° E). The focus of the study is 1951, 61 years ago, or about 5 1/2 solar cycles ago. The ionosonde was a model C-3 designed by the National Bureau of Standards (NBS). Ionograms were taken at approximately six per hour, but not uniformly spaced in time. These were recorded on an extended frame 35 mm film. Between 2-4 weeks of ionograms were recorded on a single film reel. These films were archived at the NOAA's National Geophysical Data Center (NGDC), also known as a World Data Center . Over the past five years, NGDC has been able to digitize several months from selected years of these films. These digitized ionogram images are the starting point for the ionospheric data analysis for this study. SEC has developed an image processing technique called the Expert System for Ionogram Reduction (ESIR), which has been patented [Sojka et al. 2009]. This software was developed specifically to recognize and invert an ionogram from a photographic image, producing an equivalent ionospheric electron density profile. The recognition of both virtual height and frequency axes in these ionogram photos is discussed. We demonstrate how we can validate and calibrate these scales independent of the ionosonde's virtual height and frequency markings. Examples during several months of 1951 of the automated ESIR ionogram reduction will be provided. These examples will be presented in the context of how the mid-latitude ionosphere over Washington DC in 1951 compares with the present-day ionosphere. Limitations in the data extraction are discussed from a point of view of how they might affect confidence in the inferred long-term trends in the ionosphere. Reference: Sojka J. J., D. C. Thompson, D. D. Rice (2009

  18. Effect of topical anti-Streptococcus mutans IgY gel on quantity of S. mutans on rats' tooth surface.

    PubMed

    Bachtiar, Endang W; Afdhal, Anggraeni; Meidyawati, Ratna; Soejoedono, Retno D; Poerwaningsih, Erni

    2016-06-01

    This study aims to evaluate the effect of anti-Streptococcus mutans IgY gel on quantity of S. mutans on rats' tooth surface. Sprague Dawley rats were exposed intra-orally with S. mutans Xc and were fed a caries-inducing diet 2000. The 24 rats were divided into four groups: group A had their teeth coated with IgY gel; group B received sterilized water as a control; group C had their teeth coated with IgY gel starting on the 29(th) day; and group D had their teeth coated with a gel without IgY. Plaque samples were swabbed from the anterior teeth for S. mutans colony quantification, and saliva was collected to measure immunoreactivity by enzyme-linked immunosorbent assay. The results indicated that the quantity of S. mutans in rats treated with IgY gel showed significant difference compared with the controls. After coating with IgY anti-S. mutans gel, the mean immunoreactivity in rat saliva was higher than that of the no treatment group. In conclusion, topical application with anti-S. mutans IgY gel reduced the quantity of S. mutans on the tooth surface. PMID:27352970

  19. Passive immunization with hyperimmune egg-yolk IgY as prophylaxis and therapy for poultry diseases--A review.

    PubMed

    Gadde, U; Rathinam, T; Lillehoj, Hyun S

    2015-12-01

    Passive immunization with pathogen-specific egg yolk antibodies (IgY) is emerging as a potential alternative to antibiotics for the treatment and prevention of various human and animal diseases. Laying hens are an excellent source of high-quality polyclonal antibodies, which can be collected noninvasively from egg yolks. The use of IgY offers several advantages in that it is environmentally friendly, nontoxic, and reduces the numbers of animals required for antibody production. This paper reviews the use of IgY antibodies in the treatment and prevention of enteric pathogen infections in poultry. Brief descriptions of the production, structure, and properties of IgY are also presented. Some limitations of the technology and future perspectives are discussed. PMID:26568433

  20. IGY+50, the IPY, and the electronic Geophysical Year (eGY)

    NASA Astrophysics Data System (ADS)

    Barton, C.; Baker, D. N.

    2004-12-01

    During the International Geophysical Year (1957-1958), member countries established geophysical observatories around the world. These nations were pursuing major IGY objectives - to collect geophysical data as widely as possible and to provide free access to these data for all scientists around the globe. By the beginning of the 21st century, we have achieved an unparalleled ability to acquire data and attained a good understanding of traditional regions - the troposphere, the magnetosphere, and other such "spheres". Much of the new and important science now is coming from the study of the boundaries between these regions and of coupling between geophysical domains. Thus, we need to make data available in a readily accessible form and in much greater quantities to a wider range of scientists than ever before. Several major international initiatives - notably the International Polar Year - have been proposed to commemorate and to follow on from the original IGY. As an important part of IGY+50, an International Union of Geodesy and Geophysics (IUGG) task group has recognized that a key achievement of the IGY was the establishment of a system of data centers and physical observatories. It has been agreed that for the 50th anniversary of IGY, scientific societies should promote the establishment of a system of Virtual Observatories. This can provide a forward impetus to geophysics n this century similar to that provided by the IGY fifty years ago. The proposed electronic Geophysical Year (eGY) concept embraces all available and upcoming geophysical data (e.g., atmospheric, geomagnetic, gravity, ionospheric, magnetospheric, etc.) and would organize them into a series of virtual geophysical observatories "deployed" in cyberspace. This concept implies a free access to all available data through the Internet and World Wide Web, taking advantage of existing networking hardware and software technologies (e.g., Internet, XML, Semantic Web, etc.). The eGY can be smoothly

  1. The chicken yolk sac IgY receptor, a mammalian mannose receptor family member, transcytoses IgY across polarized epithelial cells.

    PubMed

    Tesar, Devin B; Cheung, Evelyn J; Bjorkman, Pamela J

    2008-04-01

    In mammals the transfer of passive immunity from mother to young is mediated by the MHC-related receptor FcRn, which transports maternal IgG across epithelial cell barriers. In birds, maternal IgY in egg yolk is transferred across the yolk sac to passively immunize chicks during gestation and early independent life. The chicken yolk sac IgY receptor (FcRY) is the ortholog of the mammalian phospholipase A2 receptor, a mannose receptor family member, rather than an FcRn or MHC homolog. FcRn and FcRY both exhibit ligand binding at the acidic pH of endosomes and ligand release at the slightly basic pH of blood. Here we show that FcRY expressed in polarized mammalian epithelial cells functioned in endocytosis, bidirectional transcytosis, and recycling of chicken FcY/IgY. Confocal immunofluorescence studies demonstrated that IgY binding and endocytosis occurred at acidic but not basic pH, mimicking pH-dependent uptake of IgG by FcRn. Colocalization studies showed FcRY-mediated internalization via clathrin-coated pits and transport involving early and recycling endosomes. Disruption of microtubules partially inhibited apical-to-basolateral and basolateral-to-apical transcytosis, but not recycling, suggesting the use of different trafficking machinery. Our results represent the first cell biological evidence of functional equivalence between FcRY and FcRn and provide an intriguing example of how evolution can give rise to systems in which similar biological requirements in different species are satisfied utilizing distinct protein folds. PMID:18256279

  2. IgY stability in eggs stored at room temperature or at +4°C.

    PubMed

    Nilsson, E; Stålberg, J; Larsson, A

    2012-01-01

    1. The aim of this study was to investigate the stability of immunoglobulin-Y (IgY) in stored eggs from immunised hens. 2. Eggs from individual hens were randomised and stored for up to one month at room temperature, or for up to 6 months at +4°C. IgY was extracted from the egg yolks and the antibody activities were tested by ELISA. 3. There was no significant reduction in antibody titres with egg storage under these conditions. 4. Egg yolks of immunised chickens provide an inexpensive source of large amounts of polyclonal antibodies for use in immunotherapy and immunoassays. By collecting eggs from different immunised hens and pooling their yolks, it should be possible to reduce batch-to-batch variation. PMID:22404803

  3. A novel isolation method for hen egg yolk antibody, "IgY".

    PubMed

    Hatta, H; Kim, M; Yamamoto, T

    1990-10-01

    A method for isolation of egg yolk immunoglobulin, IgG, a livetin protein, was investigated. Several natural gums (carrageenan and xanthan gum) were found to be effective for removal of yolk lipoprotein as a precipitate. The effect was pronounced with lambda-carrageenan and the lipid content in the supernatant after removal of the resulting precipitate was less than 0.4% of that of egg yolk. IgY remained in this supernatant, with a yield of 86%, and was isolated by chromatography on a column of DEAE-Sephacel followed by salting-out with sodium sulfate. IgY thus isolated was almost pure (98%) and the yield was 70 to 100 mg per egg. PMID:1368596

  4. Biological and Immunogenicity Property of IgY Anti S. mutans ComD

    PubMed Central

    Bachtiar, E.W.; Bachtiar, B.M.; Soejoedono, R.D.; Wibawan, I.W.; Afdhal, A.

    2016-01-01

    Objective: This study aims to elucidate the effect of IgY anti ComD on the biological properties of Streptococcus mutans. (S. mutans) ComD is an interspecies quorum-sensing signaling receptor that plays an important role in biofilm formation by S. mutans. Materials and Methodology: Egg yolk IgY was produced by the immunization of chickens with a DNA vaccine containing the ComD DNA coding region. We evaluated the effect of the antibody on biofilm formation by S. mutans isolated from subjects with or without dental caries. We also assessed the immunoreactivity of the antibody against all isolates, and analyzed the protein profile of S. mutans by SDS-PAGE. Results: The ComD antibody was successfully induced in the hens’ eggs. It inhibited biofilm formation by all S. mutans isolates. In addition, the expression of some protein bands was affected after exposure to the antibody. Conclusion: IgY anti-S. mutans ComD reduces biofilm formation by this bacterium and alters the protein profile of S. mutans. PMID:27386013

  5. Protective effects of chicken egg yolk antibody (IgY) against experimental Vibrio splendidus infection in the sea cucumber (Apostichopus japonicus).

    PubMed

    Li, Xiaoyu; Jing, Kailin; Wang, Xitao; Li, Yuan; Zhang, Meixia; Li, Zhen; Xu, Le; Wang, Lili; Xu, Yongping

    2016-01-01

    Vibrio splendidus is one of the most harmful pathogens associated with skin ulceration syndrome in the sea cucumber (Apostichopus japonicus) due to its high virulence and frequency of appearance. The objective of this study was to determine the effectiveness of chicken egg yolk antibody (IgY) against V. splendidus infection in the sea cucumber. Whole V. splendidus cells were used as an immunogen to immunize 20 White Leghorn hens (25 weeks old). IgY was produced from egg yolks obtained from these immunized hens using water dilution, two-step salt precipitation and ultrafiltration. The purity of the IgY produced was approximately 83%. Enzyme-linked Immunosorbent Assay indicated a high specificity for IgY with a maximum antibody titer of 320,000. The growth of V. splendidus in liquid medium was significantly inhibited by IgY in a dose-dependent manner at concentrations ranging from 1 to 10 mg/mL. The protective effects of IgY were evaluated in sea cucumber by intraperitoneally injecting anti-V. splendidus IgY antibodies (10 mg/mL) or immersing the sea cucumber in aqueous IgY (1 g/L) after an intraperitoneal injection of V. splendidus. Intraperitoneal injection resulted in an 80% survival while immersion resulted in a 75% survival during the 11-day experimental period. The survival rates were significantly higher than the positive control and the non-specific IgY group (P < 0.05). As well, the bacterial burden in the respiratory tree, intestine and coelomic liquid was significantly (P < 0.05) lower in sea cucumber treated with specific IgY than those treated with non-specific IgY. The phagocytosis of coelomocytes for V. splendidus in the presence of specific IgY was significantly (P < 0.05) higher than that obtained with non-specific IgY or without IgY, suggesting that specific IgY enhanced phagocytic activity. The current work suggests that specific IgY has potential for protecting sea cucumbers against V. splendidus infection. PMID:26592708

  6. Moving Beyond IGY: An Electronic Geophysical Year (eGY) Concept

    NASA Astrophysics Data System (ADS)

    Baker, D. N.; Barton, C. E.; Rodger, A. S.; Thompson, B. J.; Fraser, B.; Papitashvili, V.

    2003-12-01

    During the International Geophysical Year (1957-1958), member countries established many new geophysical observatories pursuing the major IGY objectives - to collect geophysical data as widely as possible and to provide free access to these data for all scientists around the globe. Today, geophysics has attained a rather good understanding within traditional regions, i.e., the atmosphere, ionosphere, magnetosphere, and other such geospheres. At the same time, it has become clear that much of the new and important science is coming from the studies of interfaces and coupling between geospheres. Thus, if geophysical data are made `'transparently'' available to a much wider range of scientists and students than to those who do the observations, then new and exciting discoveries can be expected. An International Association of Geomagnetic and Aeronomy (IAGA) task force, recognizing that a key achievement of the IGY was the establishment of a worldwide system of data centers and physical observatories, proposes that for the 50th anniversary of IGY, the worldwide scientific community should endorse and promote an electronic Geophysical Year (eGY) initiative. The proposed eGY concept would both commemorate the IGY in 2007-2008 and provide a forward impetus to geophysics in 21st century, similar to that provided by the IGY fifty years ago. The IAGA task force strongly advocates: (1) Securing permission and release of existing data; (2) Creating access to information; and (3) Conversion of relevant analog data to digital form. The eGY concept embraces all available and upcoming geophysical data (e.g., atmospheric, ionospheric, geomagnetic, gravity, etc.) through the establishment of a series of virtual geophysical observatories now being `'deployed'' in cyberspace. The eGY concept is modern, global, and timely; it is attractive, pragmatic, and affordable. The eGY is based on the existing and continually developing computing/networking technologies (e.g., XML, Semantic Web

  7. Transient developmental expression of IgY and secretory component like protein in the gut of the axolotl (Ambystoma mexicanum).

    PubMed

    Fellah, J S; Iscaki, S; Vaerman, J P; Charlemagne, J

    1992-01-01

    We previously reported that a primitive vertebrate, the Mexican axolotl (Amphibian, Urodela) synthesizes two classes of immunoglobulins. IgM are present in serum early in the development, and represent the bulk of specific antibody synthesis after an antigenic challenge. IgY occur in the serum later during the development, and are relatively insensitive to immunization. We demonstrate in the present work, using immunofluorescence with specific Mabs, that IgY are expressed in the gut epithelium, as secretory molecules. Secretory IgY are well expressed in the stomach and intestinal mucosae of young animals from 1 month after hatching to the seventh month. Thereafter, IgY progressively disappear from the gut and become readily detectable in the serum of 9-month-old preadult immunologically mature animals. Axolotl IgY are closely associated in the gut to secretory component-like (SC) molecules that are well-recognized by antisera to the SC of different mammalian species. This is the first description, in a primitive tetrapode, of an immunoglobulin class that could be the physiological counterpart of mammalian IgA. PMID:1627950

  8. Reproduction of Meloidogyne incognita Race 3 on Flue-cured Tobacco Homozygous for Rk1 and/or Rk2 Resistance Genes.

    PubMed

    Pollok, Jill R; Johnson, Charles S; Eisenback, J D; Reed, T David

    2016-06-01

    Most commercial tobacco cultivars possess the Rk1 resistance gene to races 1 and 3 of Meloidogyne incognita and race 1 of Meloidogyne arenaria, which has caused a shift in population prevalence in Virginia tobacco fields toward other species and races. A number of cultivars now also possess the Rk2 gene for root-knot resistance. Experiments were conducted in 2013 to 2014 to examine whether possessing both Rk1 and Rk2 increases resistance to a variant of M. incognita race 3 compared to either gene alone. Greenhouse trials were arranged in a completely randomized design with Coker 371-Gold (C371G; susceptible), NC 95 and SC 72 (Rk1Rk1), T-15-1-1 (Rk2Rk2), and STNCB-2-28 and NOD 8 (Rk1Rk1 and Rk2Rk2). Each plant was inoculated with 5,000 root-knot nematode eggs; data were collected 60 d postinoculation. Percent galling and numbers of egg masses and eggs were counted, the latter being used to calculate the reproductive index on each host. Despite variability, entries with both Rk1 and Rk2 conferred greater resistance to a variant of M. incognita race 3 than plants with Rk1 or Rk2 alone. Entries with Rk1 alone were successful in reducing root galling and nematode reproduction compared to the susceptible control. Entry T-15-1-1 did not reduce galling compared to the susceptible control but often suppressed reproduction. PMID:27418700

  9. Reproduction of Meloidogyne incognita Race 3 on Flue-cured Tobacco Homozygous for Rk1 and/or Rk2 Resistance Genes

    PubMed Central

    Pollok, Jill R.; Johnson, Charles S.; Eisenback, J. D.; Reed, T. David

    2016-01-01

    Most commercial tobacco cultivars possess the Rk1 resistance gene to races 1 and 3 of Meloidogyne incognita and race 1 of Meloidogyne arenaria, which has caused a shift in population prevalence in Virginia tobacco fields toward other species and races. A number of cultivars now also possess the Rk2 gene for root-knot resistance. Experiments were conducted in 2013 to 2014 to examine whether possessing both Rk1 and Rk2 increases resistance to a variant of M. incognita race 3 compared to either gene alone. Greenhouse trials were arranged in a completely randomized design with Coker 371-Gold (C371G; susceptible), NC 95 and SC 72 (Rk1Rk1), T-15-1-1 (Rk2Rk2), and STNCB-2-28 and NOD 8 (Rk1Rk1 and Rk2Rk2). Each plant was inoculated with 5,000 root-knot nematode eggs; data were collected 60 d postinoculation. Percent galling and numbers of egg masses and eggs were counted, the latter being used to calculate the reproductive index on each host. Despite variability, entries with both Rk1 and Rk2 conferred greater resistance to a variant of M. incognita race 3 than plants with Rk1 or Rk2 alone. Entries with Rk1 alone were successful in reducing root galling and nematode reproduction compared to the susceptible control. Entry T-15-1-1 did not reduce galling compared to the susceptible control but often suppressed reproduction. PMID:27418700

  10. [Agglutination of hen egg-yolk immunoglobulins (IgY) against Salmonella enterica, serovar enteritidis].

    PubMed

    Terzolo, H R; Sandoval, V E; Caffer, M I; Terragno, R; Alcain, A

    1998-01-01

    Two groups of 6 laying hens were used to produce IgY. In the vaccinated group (V), hens were injected by intramuscular route with two doses of a Salmonella enterica serovar Enteritidis bacterin at 20-day interval. In the control group (T) hens remained unvaccinated. Four IgY extractions were performed on the egg production of both groups. The first two extractions were carried out using the yolks obtained from the eggs produced during the 4th and 5th post-vaccination week (extracts 1V and 1T) and the other two using the ones from the 6th, 7th and 8th week (2V and 2T). Starting from the extracts 1V and 1T other products were obtained by freezing-thawing (1V-A and 1T-A) and simple (1V-B and 1T-B) or double (1V-C and 1T-C) flow capillary dialysis concentration. All these products were compared using an ELISA test specific for the detection of chicken antibodies against flagellar antigens of S. Enteritidis. In this test, V extracts were positive whereas T extracts were negative. The extract 1V was more positive than the extract 2V. The extract 1V-C was the most positive and was therefore selected to be used as an antiserum in the agglutination tests. This extract contained 1.9 g/dl of total proteins, 0.028 g/dl of triglycerides and 0.012 g/dl of cholesterol and showed an electrophoretic pattern characteristic of IgY. The 1T-C extract was used as a negative control in the agglutination tests. Slide somatic and tube flagellar agglutination tests were simultaneously carried out using both IgY extracts and a standard rabbit anti-Salmonella (IgG) sera. Overall 367 strains from the Enterobacteriaceae family were tested together with two other strains belonging to the Vibrionaceae family. The 1V-C extract specifically agglutinated S. Enteritidis strains in the same way as the rabbit sera. This extract also agglutinated other Salmonella strains antigenically related to S. Enteritidis. Salmonella which did not share somatic or flagellar antigens with S. Enteritidis, other

  11. Ulcer disease prophylaxis in koi carp by bath immersion with chicken egg yolk containing anti-Aeromonas salmonicida IgY.

    PubMed

    Gan, Hongjian; He, Haiwen; Sato, Atsushi; Hatta, Hajime; Nakao, Miki; Somamoto, Tomonori

    2015-04-01

    Ulcer disease, caused by atypical Aeromonas salmonicida, is a serious concern in ornamental koi carp, because it induces skin ulceration, disfiguring ornamental fish and causing economic loses. The present study aimed to establish a novel prophylaxis with chicken egg yolk immunoglobulin, IgY, against ulcer disease and to assess its feasibility in the ornamental fish industry. Addition of egg yolk powder containing anti-A. salmonicida IgY to rearing water provided significant protection against an A. salmonicida bath infection, whereas administration of non-specific IgY did not. Consecutive immersion of fish into rearing water containing specific IgY completely prevented ulcer disease resulting from cohabitation infection, indicating that this prophylaxis could prevent infection from such type of contact. Thus, passive immunization induced by immersing fish into aquarium water containing specific IgY is a prospective prophylaxis against diseases caused by pathogens that invade the skin and gills. PMID:25687817

  12. Production and characterization of egg yolk antibody (IgY) against recombinant VP8-S2 antigen.

    PubMed

    Nasiri, K; Nassiri, M R; Tahmoorespur, M; Haghparast, A; Zibaee, S

    2016-01-01

    Bovine Rotavirus and Bovine Coronavirus are the most important causes of diarrhea in newborn calves and in some other species such as pigs and sheep. VP8 subunit of rotavirus is the major determinant of the viral infectivity and neutralization. Spike glycoprotein of coronavirus is responsible for induction of neutralizing antibody response. Studies showed that immunoglobulin of egg yolk (IgY) from immunized hens has been identified to be a convenient source for specific antibodies for using in immunotherapy and immunodiagnostic to limit the infections. In this study, chimeric VP8-S2 gene was designed using by computational techniques. The chimeric VP8-S2 gene was cloned and sub-cloned into pGH and pET32a (+) vectors. Then, recombinant pET32a-VP8-S2 vector was transferred into E. coli BL21 CodonPlus (DE3). The expressed protein was purified by Ni-NTA chromatography column. Hens were immunized with the purified VP8-S2 protein three times. IgY was purified from egg yolks using polyethylene glycol precipitation method. Activity and specificity of anti-VP8-S2 IgY were detected by dot-blotting, Western-blotting and indirect ELISA. We obtained anti-VP8-S2 IgY by immunizing hens with the recombinant VP8-S2 protein. The anti-VP8-S2 IgY was showed to bind specifically to the chimeric VP8-S2 protein by dot-blotting, Western-blotting analyses and indirect ELISA. The result of this study indicated that such construction can be useful to investigate as candidates for development of detection methods for simultaneous diagnosis of both infections. Specific IgY against the recombinant VP8-S2 could be recommended as a candidate for passive immunization against bovine rotavirus and bovine coronavirus. PMID:27487500

  13. View of the IGY from a high school physics classroom (1959-1961)

    NASA Astrophysics Data System (ADS)

    Michels, D.

    Convening a worldwide collaborative scientific effort can be expected to produce many advantages including improved focus and definition of scientific goals through negotiated agreements, the coordination of various investigative approaches, and synchro nization of observations at widespread sites and with diverse techniques. Collateral advantages might include improved leveraging of national resources, especially for small or less developed countries. Not to be overlooked, however, is the immense potential for gains in public enthusiasm, support and educational opportunities. This talk will focus on personal experiences and retrospective observations of secondary school physics classes taught during the IGY extension years, 1959-1961 as new and unexpected images and data flooded into world view and every physics lesson could be linked somehow, to the morning's headlines. Twenty-five years later our Nation, it seems, is again ripe for a sea change in physical science education, and we find ourselves organizing traditional physics topics around the fascinating framework of Sun-Earth Connection physics. We will discuss educational lessons learned from the IGY and briefly mention plans for applying them in the IHY.

  14. Production of anti-SAG1 IgY antibody against Toxoplasma gondii parasites and evaluation of antibody activity by ELISA method.

    PubMed

    Cakir-Koc, Rabia

    2016-08-01

    Chicken egg yolk antibody, also known as immunoglobulin Y (IgY), is the predominant class of serum immunoglobulin in birds. IgY has many advantages over mammalian antibodies, such as enhanced immunogenicity conserved mammalian proteins exhibited in birds due to their phylogenetic distance, non-invasive rapid, and economical collection system. However, there are limited studies about IgY production against Toxoplasma, which is a worldwide veterinary and public health problem. In this study, the production of specific IgY antibodies against the surface antigen 1 (SAG1) protein of Toxoplasma gondii and the determination of antibody activity via the enzyme-linked immunosorbent assay (ELISA) method were conducted. According to ELISA, Western blot, and NanoDrop results, specific and higher amounts of IgY antibody against SAG1 were obtained with this study. Considering the advantages of IgY and importance of SAG1 for the diagnosis of toxoplasmosis, it is expected that anti-SAG1 IgY will play an increasing role and gain commercial value in research, diagnostics, and immunotherapy against toxoplasmosis in the future. PMID:27079459

  15. Induction of Passive Immunity in Broiler Chickens Against Eimeria acervulina by Hyperimmune Egg Yolk IgY

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The protective effect of hyperimmune IgY fraction of egg yolk (SC) prepared from hens hyperimmunized with multiple species of Eimeria oocysts, on experimental coccidiosis was evaluated in young broilers. Chickens were continuously fed from hatch with a standard diet containing SC or a non-suppleme...

  16. Induction of passive immunity in broiler chickens against Eimeria acervulina by hyperimmune egg yolk IgY

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The protective effect of hyperimmune IgY fraction of egg yolk (SC) prepared from hens hyperimmunized with multiple species of Eimeria oocysts, on experimental coccidiosis was evaluated in young broilers. Chickens were continuously fed from hatch with a standard diet containing SC or a non-suppleme...

  17. Inhibitory effects of rHP-NAP IgY against Helicobacter pylori attachment to AGS cell line.

    PubMed

    Borhani, Katayoun; Mohabati Mobarez, Ashraf; Khabiri, Ali Reza; Behmanesh, Mehrdad; Khoramabadi, Nima

    2016-08-01

    Helicobacter pylori is a major human pathogen related to gastric adenocarcinoma and gastroduodenal diseases. Treatment of H. pylori infections is complicated by the rise of antibiotic resistance, necessitating investigation of alternative therapies. One such alternative is passive immunization by oral administration of antibacterial immunoglobulin. In the present study, chicken immunoglobulin (IgY) was used for passive immunotherapy against a major virulence factor of H. pylori, namely recombinant HP-Nap protein. Recombinant HP-Nap was prepared and used to immunize hens. IgY was purified from the eggs by polyethylene glycol precipitation method with a total IgY-HP-NAP yield of 30 mg per egg. The inhibitory effect of specific IgY on H. pylori attachment was investigated in AGS cell line infected by the bacteria. The results demonstrate the potent effect of IgY- HP-NAP in inhibition of H. pylori attachment to the AGS cells. PMID:27265677

  18. Isolation and Characterization of IgM and IgY Antibodies from Plasma of Magellanic Penguins (Spheniscus magellanicus).

    PubMed

    Bizelli, Camila C; Silva, A Sandriana R; da Costa, Jessica D; Vanstreels, Ralph E T; Atzingen, Marina V; Santoro, Marcelo L; Fernandes, Irene; Catão-Dias, José L; Faquim-Mauro, Eliana L

    2015-03-01

    Infectious diseases such as aspergillosis, avian malaria, and viral infections are significant threats to the conservation of penguins, leading to morbidity and mortality of these birds both in captivity and in the wild. The immune response to such infectious diseases is dependent on different mechanisms mediated by cells and soluble components such as antibodies. Antibodies or immunoglobulins are glycoproteins that have many structural and functional features that mediate distinct effector immune functions. Three distinct classes of antibodies have been identified in birds: immunoglobulin A (IgA), immunoglobulin M (IgM), and immunoglobulin Y (IgY). In this study we aim to establish an efficient laboratory method to obtain IgM and IgY antibodies from plasma samples of healthy adult Magellanic penguins (Spheniscus magellanicus). The protocol was developed combining plasma delipidation, sequential precipitation with caprylic acid and ammonium sulfate, and size-exclusion chromatography. The efficiency of the protocol and the identity of the purified IgM and IgY antibodies were confirmed through enzyme-linked immunosorbent assay, Western blotting, one-dimensional and two-dimensional polyacrylamide gel electrophoresis, and lectin binding assay. Structural and physicochemical properties of IgM and IgY from Magellanic penguins were consistent with those of other avian species. This purification protocol will allow for more detailed studies on the humoral immunity of penguins and for the development of high specificity serologic assays to test Magellanic penguins for infectious pathogens. PMID:26292539

  19. Conceptualizing Relations between Instructional Guidance Infrastructure (IGI) and Teachers' Beliefs about Mathematics Instruction: Regulative, Normative, and Cultural-Cognitive Considerations

    ERIC Educational Resources Information Center

    Hopkins, Megan; Spillane, James P.

    2015-01-01

    Scholars have become increasingly interested in what is often referred to as the instructional guidance infrastructure (IGI). Research has identified the characteristics of infrastructures that make them more or less influential in guiding teachers' instruction, such as alignment, authority, and prescriptiveness. Although these are important, a…

  20. Development of indirect competitive ELISA using egg yolk-derived immunoglobulin (IgY) for the detection of Gentamicin residues.

    PubMed

    He, Jinxin; Hu, Jianjun; Thirumalai, Diraviyam; Schade, Ruediger; Du, Enqi; Zhang, Xiaoying

    2016-01-01

    Gentamicin (Gent) is an aminoglycoside antibiotic being used in livestock sector. Gent residues could cause some genetic disorders by nonsense mutations. This study aimed to develop IgY-based ELISA for the detection of Gent in animal products. Gent was conjugated with Bovine serum albumin (BSA) by carbodiimide method for further immunization in the laying chickens. PEG-6000 extraction method was employed to extract IgY from the egg yolk. The titer of anti-Gent-IgY attained the peak of 1:256,000 after the 5(th) booster immunization. Checkerboard titration confirmed that, anti-Gent IgY in 1:2,000 dilution could give an Optical Density (OD) 1.0 at 2 µg mL(-1) of Gent-OVA coating concentration. IgY-based indirect competitive ELISA (Ic-ELISA) showed that, the IC50 value of anti-Gent IgY was 2.69 ng mL(-1) and regression curve equation was y = -16.27x + 56.97 (R(2) = 0.95, n = 3), confirming that, the detection limit (LOD, IC10 value) was 0.01 ng mL(-1). Recoveries from fresh milk, pork and chicken samples were ranged from 69.82% to 94.32%, with relative standard deviation lower than 10.88%. Our results suggested that generated anti-Gent IgY antibodies can be used in routine screening analysis of Gent residues in food samples. PMID:26513166

  1. Effect of Chicken Egg Yolk Antibodies (IgY) against Diarrhea in Domesticated Animals: A Systematic Review and Meta-Analysis

    PubMed Central

    Diraviyam, Thirumalai; Zhao, Bin; Wang, Yuan; Schade, Ruediger; Michael, Antonysamy; Zhang, Xiaoying

    2014-01-01

    Background IgY antibodies are serum immunoglobulin in birds, reptiles and amphibians, and are transferred from serum to egg yolk to confer passive immunity to their embryos and offspring. Currently, the oral passive immunization using chicken IgY has been focused as an alternative to antibiotics for the treatment and control of diarrhea in animals and humans. This systematic review was focused to determine the effect of IgY in controlling and preventing diarrhea in domesticated animals including Piglets, Mice, Poultry and Calves. Methods and Results Previous research reports focused on treatment effect of Chicken IgY against diarrhea were retrieved from different electronic data bases (MEDLINE, EMBASE, SPRINGER-LINK, WILEY, AGRICOLA, MEDWELL Journals, Scientific Publish, Chinese articles from Core periodicals in 2012). A total of 61 studies in 4 different animal classes met the inclusion criteria. Data on study characteristics and outcome measures were extracted. The pooled relative risk (RR) of 49 studies of different animals [Piglets – 22; Mice – 14; Poultry – 7 and Calves – 6] in meta-analyses revealed that, IgY significantly reduced the risk of diarrhea in treatment group when compare to the placebo. However, the 95% confidence intervals of the majority of studies in animal class piglets and calves embrace RR of one. The same results were obtained in sub group analyses (treatment regiment – prophylactic or therapeutic; pathogen type – bacterial or viral). Perhaps, this inconsistency in the effect of IgY at the individual study level and overall effect measures could be influenced by the methodological heterogeneity. Conclusion The present systematic review (SR) and meta-analysis demonstrated the beneficial effect of IgY. This supports the opinion that IgY is useful for prophylaxis and treatment. However, more intensive studies using the gold standard animal experiments with the focus to use IgY alone or in combination with other alternative

  2. IGY to IPY, the U.S. Antarctic oversnow and airborne geophysical-glaciological research program from 1957 to 1964 from the view of a young graduate student

    USGS Publications Warehouse

    Behrendt, John C.

    2007-01-01

    When 12 countries established scientific stations in Antarctica for the 1957-58 (IGY), the Cold War was at its height, seven countries had made claims in Antarctica, and the Antarctic Treaty was in the future. The only major field project of the U.S. IGY Antarctic program was series of oversnow traverses, starting in 1957, making seismic reflection ice soundings (and other geophysical measurements) and glaciological studies. The U.S.S.R. and France made similar traverses coordinated through the IGY. Although geology and topographic mapping were not part of the IGY program because of the claims issue and the possibility of mineral resources, the oversnow traverse parties did geologic work, during which unknown mountains were discovered. The oversnow traverses continued through 1966 and resulted in an excellent first approximation of the snow surface elevation, ice thickness and bed topography of Antarctica, as well as the mean annual temperature of that era and snow accumulation.

  3. SymRK and the nodule vascular system

    PubMed Central

    Sanchez-Lopez, Rosana; Jáuregui, David; Quinto, Carmen

    2012-01-01

    Symbiotic legume-rhizobia relationship leads to the formation of nitrogen-fixing nodules. Successful nodulation depends on the expression and cross-talk of a batttery of genes, among them SymRK (symbiosis receptor-like kinase), a leucine-rich repeat receptor-like kinase. SymRK is required for the rhizobia invasion of root hairs, as well as for the infection thread and symbiosome formation. Using immunolocalization and downregulation strategies we have recently provided evidence of a new function of PvSymRK in nodulation. We have found that a tight regulation of PvSymRK expression is required for the accurate development of the vascular bundle system in Phaseolus vulgaris nodules. PMID:22580688

  4. Apoptotic Killing of Breast Cancer Cells by IgYs Produced Against a Small 21 Aminoacid Epitope of the Human TRAIL-2 Receptor.

    PubMed

    Amirijavid1, Shaghayegh; Entezari, Maliheh; Movafagh, Abolfazl; Hashemi, Mehrdad; Mosavi-Jarahi, Alireza; Dehghani, Hossein

    2016-01-01

    TRAIL, tumor necrosis factor (TNF)-related apoptosis-inducing ligand belongs to one of important cytokine superfamilIES, tumor necrosis factor (TNFα) . TRAIL-2 receptor agonists activate several cell signaling pathways in cells in different manners and could lead to apoptosis or necrosis. Agonistic egg yolk antibodies like IgY which have been developed in a selective manner could activate TRAIL death receptors such as TRAIL-2 (DR5) and thus apoptosis signaling. We here investigated induction of apoptosis in human breast cancer cells (MCF7 cell line) by an IgY produced against an 21 aminoacid epitope of the human TRAIL-2 receptor. As the first step a small peptide of 21 aminoacids choosen from the extracellular domain of DR5 protein was produced with a peptide synthesizer. After control assays and confirmation of the correct amino acid sequence, it was injected to hens immunized to achieve high affinity IgYs. At the next step, the produced IgYs were extracted and examined for specificity against DR5 protein by ELISA assay. Subsequently, the anticancer effect of such IgYs was determined by MTT assay in the MCF7 human breast cancer cell line. The produced peptides successfully immunized hens and the produced antibodies which accumulated in egg yolk specifically recognized the DR5 protein. IgYs exerted significant toxicity and killed MCF7 cells as shown by MTT assay. PMID:27165241

  5. Chicken egg yolk antibodies (IgY) modulate the intestinal mucosal immune response in a mouse model of Salmonella typhimurium infection.

    PubMed

    Li, Xiaoyu; Yao, Ying; Wang, Xitao; Zhen, Yuhong; Thacker, Philip A; Wang, Lili; Shi, Ming; Zhao, Junjun; Zong, Ying; Wang, Ni; Xu, Yongping

    2016-07-01

    This study determined the effects of chicken egg yolk antibodies (IgY) on immune responses in the intestinal mucosal of mice infected with Salmonella typhimurium. Sixty, 28-day-old mice were divided into 4 groups and treated with streptomycin or sterile water for 2days followed by 1day without treatment. The control group was unchallenged whereas the mice in the other three groups were treated twice with 10(9)CFUmL(-1)S. typhimurium. For the next 3days, control mice continued to receive no treatment whereas the mice in the remaining three groups were orally administered with 20mgmL(-1) of specific IgY, 20mgmL(-1) of nonspecific IgY or PBS. S. typhimurium activated gut-associated lymphoid tissue, increasing the release of IFN-γ and TNF-α in the mucosa and increased the number of activated T-lymphocytes and cytotoxic T-γδ. Specific IgY attenuated the increase in IFN-γ and TNF-α and the decrease in IL-10. S. typhimurium induced mobilization of CD8(+) and CD8(+) TCRγδ T cells in the epithelium and CD4(+) and CD8(+) T cells in the lamina propria reflecting an inflammatory process that was attenuated by IgY. These results suggest that specific IgY modulates intestinal mucosal immune responses during a S. typhimurium infection. PMID:27214338

  6. IgY against enterotoxigenic Escherichia coli administered by hydrogel-carbon nanotubes composites to prevent neonatal diarrhoea in experimentally challenged piglets.

    PubMed

    Alustiza, Fabrisio; Bellingeri, Romina; Picco, Natalia; Motta, Carlos; Grosso, Maria C; Barbero, Cesar A; Acevedo, Diego F; Vivas, Adriana

    2016-06-14

    In previous studies, the applicability of polymeric hydrogels for the protection of egg yolk immunoglobulin (IgY) against simulated gastric conditions was established. Thereafter, the performance of the hydrogels was improved with the addition of chitosan wrapped carbon nanotubes and the in vitro toxicity for porcine intestinal cells of these nanocomposites was assessed. The objective of the present study was to evaluate in vivo the protective efficacy of the nanocomoposite matrix for IgY when the immunoglobulin is used against enterotoxigenic Escherichia coli (ETEC) in challenged piglets. Groups of piglets orally challenged with 10(11)CFU/mL of ETEC were treated with non-protected and protected IgY. The clinical response of each group was monitored and evaluated in terms of dehydration, rectal temperature, faecal consistency score and body weight gain. Blood parameters and histological aspects were also studied. The results showed that treatment of infected piglets with protected IgY reduced significantly the severity of diarrhea. Non-protected IgY group show a lower recovery rate. Blood parameters and histological aspects were normal in both groups. Collectively, these results support previous in vitro studies showing that the nanocomposites can be an effective method of IgY protection against gastric inactivation. PMID:27166825

  7. Anti-Pseudomonas aeruginosa IgY antibodies promote bacterial opsonization and augment the phagocytic activity of polymorphonuclear neutrophils.

    PubMed

    Thomsen, Kim; Christophersen, Lars; Jensen, Peter Østrup; Bjarnsholt, Thomas; Moser, Claus; Høiby, Niels

    2016-07-01

    Moderation of polymorphonuclear neutrophils (PMNs) as part of a critical defense against invading pathogens may offer a promising therapeutic approach to supplement the antibiotic eradication of Pseudomonas aeruginosa infection in non-chronically infected cystic fibrosis (CF) patients. We have observed that egg yolk antibodies (IgY) harvested from White leghorn chickens that target P. aeruginosa opsonize the pathogen and enhance the PMN-mediated respiratory burst and subsequent bacterial killing in vitro. The effects on PMN phagocytic activity were observed in different Pseudomonas aeruginosa strains, including clinical isolates from non-chronically infected CF patients. Thus, oral prophylaxis with anti-Pseudomonas aeruginosa IgY may boost the innate immunity against Pseudomonas aeruginosa in the CF setting by facilitating a rapid and prompt bacterial clearance by PMNs. PMID:26901841

  8. Growth inhibition of Staphylococcus aureus and escherichia coli strains by neutralizing IgY antibodies from ostrich egg yolk

    PubMed Central

    Tobias, Fernando Luiz; Garcia, Luize Néli Nunes; Kanashiro, Milton Masahiko; Medina-Acosta, Enrique; Brom-de-Luna, João Gato; de Almeida, Claudia Maria Costa; Azevedo Junior, Romildo Rocha; Lemos, Môsar; Vieira-da-Motta, Olney

    2012-01-01

    Ostrich raising around the world have some key factors and farming profit depend largely on information and ability of farmers to rear these animals. Non fertilized eggs from ostriches are discharged in the reproduction season. Staphylococcus aureus and Escherichia coli are microorganisms involved in animal and human diseases. In order to optimize the use of sub products of ostrich raising, non fertilized eggs of four selected birds were utilized for development of polyclonal IgY antibodies. The birds were immunized (200ug/animal) with purified recombinant staphylococcal enterotoxin C (recSEC) and synthetic recRAP, both derived from S. aureus, and recBFPA and recEspB involved in E. coli pathogenicity, diluted in FCA injected in the braquial muscle. Two subsequent immunization steps with 21 days intervals were repeated in 0,85% saline in FIA. Blood and eggs samples were collected before and after immunization steps. Egg yolk immunoglobulins were purified by precipitation with 19% sodium sulfate and 20% ammonium sulphate methodologies. Purified IgY 50μL aliquots were incubated in 850μL BHI broth containing 50μL inoculums of five strains of S. aureus and five strains of E.coli during four hours at 37°C. Growth inhibition was evaluated followed by photometry reading (DO550nm). Egg yolk IgY preparation from hiperimmunized birds contained antibodies that inhibited significantly (p<0,05) growth of strains tested. Potential use of ostrich IgY polyclonal antibodies as a diagnostic and therapeutic tool is proposed for diseased animals. PMID:24031862

  9. Antarctica and Its Ice Sheet: Knowledge Gained During the IGY/IGC

    NASA Astrophysics Data System (ADS)

    Bentley, C. R.

    2006-12-01

    At the end of World War II, the interior of Antarctica, with the exception of the mountains south of the Ross Ice Shelf, was still terra incognita. It was described simply as a nearly continuous high plateau. Even less was known about the ice thickness; the eminent glacial geologist, Richard Foster Flint, believed it "unlikely that the ice thickness exceeds 2000 feet except very locally; probably its average thickness is considerably less." Then in the late 1940's and early 1950's, seismic sounding in Greenland by the Expéditions Polaires Françaises and in Queen Maud Land by the Norwegian-British-Swedish Antarctic Expedition, 1949-52, revealed that, inland of the coastal mountains, the beds in both regions lie close to sea level. This led to a reappraisal of the Antarctic ice sheet, such that the prescient glaciologist, Robert P. Sharp, could predict, on the eve of the IGY, that "between 3000 and 4000 meters of ice will be found" in East Antarctica and that "work during IGY will establish an average thickness for Antarctic inland ice in excess of 1600 m." Seismic and gravity soundings on oversnow traverses conducted by eight countries during the IGY and the succeeding IGC showed Sharp to be basically correct, but there were major surprises, such as the vast Gamburtsev Subglacial Mountains, completely hidden by the ice in central East Antarctica, and the equally vast Byrd Subglacial Basin beneath much of the West Antarctic ice sheet, so deep that roughly half the ice in the region lies below sea level. There were major discoveries on and above the surface too, such as the huge size of the Filchner/Ronne Ice Shelf, and the very existence of the Ellsworth and Pensacola Mountains, the former including the highest peak on the continent. Further, the fundamental difference between the crustal structures of East and West Antarctica became clear. A summary paper published in 1960, looking primarily at West Antarctica where the main U.S. activity lay, could conclude that

  10. Free and open exchange of data: The legacy of IGY and the goal of eGY

    NASA Astrophysics Data System (ADS)

    Baker, D. N.

    2006-05-01

    The series of International Polar Years leading up to the International Geophysical Year (IGY) in 1957-1958 taught scientists that the free and open exchange of data between nations was cost effective and scientifically beneficial. The IGY also led to the development of a world-wide network of data centers that have facilitated and fostered research begun in the IGY. We now have achieved an unparalleled ability to acquire data and have attained a good understanding of traditional regions - the troposphere, the magnetosphere, and other such "spheres". Much of the new and important science is presently coming from the study of the boundaries between these regions and of coupling between geophysical domains. The past 15 years have seen the development of new, cost effective ways to acquire, store, and exchange data. We have the potential to expand the free and open exchange of data by allowing working scientists to access and manipulate data from large interdisciplinary data centers as well as from small, previously isolated, research groups. The key to this new technology requires adoption of a few community-developed standards for data storage and description. It has been agreed that for the 50th anniversary of IGY, scientific societies should promote the establishment of a system of Virtual Observatories. This can provide a forward impetus to geophysics in this century similar to that provided by the IGY fifty years ago. The electronic Geophysical Year (eGY) concept embraces all available and upcoming geophysical data (e.g., oceanographic, seismic, atmospheric, geomagnetic, gravity, ionospheric, magnetospheric, etc.) and would organize them into a series of virtual geophysical observatories "deployed" in cyberspace. This concept implies a free access to all available data through the Internet and World Wide Web, taking advantage of existing networking hardware and software technologies (e.g., Internet, XML, Semantic Web, etc.). The eGY can be smoothly incorporated

  11. A half century perspective on the International Geophysical Year (IGY) - A Template for the International Polar Year 2007 (IPY 2007)?

    NASA Astrophysics Data System (ADS)

    Behrendt, J. C.

    2003-12-01

    In 1956 I sailed for Antarctica to spend 18 months as a graduate student participating in geophysical-glaciological investigations, as part of the 18-month IGY. This led to a career in geophysics, which has taken me to all of the continents and oceans. As we approach the IPY 2007, the changes in technology and our understanding of the earth over the past half century are breathtaking to contemplate. Although 70 countries participated in IGY, the disciplines were restricted to geophysics. Originally the Third Polar Year, the name was changed to IGY in 1952, at the suggestion of Sydney Chapman. The geographical area comprised the entire earth. The highest priority was given to "problems requiring concurrent synoptic observations at many points involving cooperative observations by many stations." One category was reserved for research on topics such as ocean levels, weather patterns, and the distribution of glacier ice "to establish basic information for subsequent comparison at later epochs." IPY 2007 seems such an epoch. A major international efforts was concentrated in Antarctica, although only 12 counties participated. Glaciology, seismology, auroral studies, ionospheric soundings, magnetic field measurements, and other solar-terrestrial, and meteorological observations comprised the scientific station activities. The only major field activities away from the stations were the oversnow geophysical-glaciological traverses, which made seismic measurements of ice thickness and other ice properties; gravity and magnetic anomaly profiles; and determination of snow accumulation and mean annual temperature. The most intensive of the oversnow traverse programs were those of the U.S. and USSR. Geology and topographic mapping were excluded from the Antarctica because of potential complications due to territorial claims and the possibility of mineral resource discoveries. Despite this, significant geologic findings, such as the discovery of the Dufek intrusion, were made by

  12. An unintended consequence of the IGY: Eisenhower, Sputnik, the Founding of NASA

    NASA Astrophysics Data System (ADS)

    Launius, Roger D.

    2010-07-01

    On October 4, 1957, the Soviet Union launched the first Earth-circling artificial satellite and the United States responded by taking numerous actions aimed at "remediating" a Cold War crisis. This included the establishment of a separate civilian space agency charged with the conduct of an official program of scientific and technological space exploration, consolidation of Department of Defense space activities, the passage of the National Defense Education Act, the creation of a Presidential Science Advisor, and a host of lesser actions. The politics of these changes is fascinating, and has been interpreted as an appropriate political response to a unique crisis situation. Interest groups, all for differing reasons, prodded national leaders to undertake large-scale efforts, something the president thought unnecessarily expensive and once set in place impossible to dismantle. But was the Sputnik crisis truly a crisis in any real sense? Was it made into one by interest groups who used it for their own ends? This paper will trace briefly some of the major themes associated with the IGY and Sputnik and describe the political construction of the crisis as it emerged in 1957-1958. It will also discuss something about the transformation of federal science and technology that took place in response to this "crisis" and how it set in train a series of processes and policies that did not unravel until the end of the Cold War.

  13. An Unintended Consequence of the IGY: Eisenhower, Sputnik, and the Founding of NASA

    NASA Astrophysics Data System (ADS)

    Launius, Roger

    2007-04-01

    In October 1957 the Soviet Union launched the first Earth-circling artificial satellite and the crisis that resulted led to numerous actions in the United States aimed at ``remediating'' a Cold War crisis. This included the establishment of a separate civilian space agency charged with the conduct of an official program of scientific and technological space exploration, consolidation of Department of Defense space activities, the passage of the National Defense Education Act, the creation of a Presidential Science Advisor, and a host of lesser actions. The politics of these changes is fascinating, and has been interpreted as an appropriate political response to a unique crisis situation. Interest groups, all for differing reasons, prodded national leaders to undertake large-scale efforts, something the president thought unnecessarily expensive and once set in place impossible to dismantle. But was the Sputnik crisis truly a crisis in any real sense? Was it made into one by interest groups who used it for their own ends? This paper will trace briefly some of the major themes associated with the IGY and Sputnik and describe the political construction of the crisis as it emerged in 1957-1958. It will also discuss something about the transformation of federal science and technology that took place in the aftermath of the ``crisis'' and how it set in train a series of processes and policies that did not unravel until the end of the Cold War.

  14. IGY@50: A Revolution in Opportunities for Public Engagement and Education

    NASA Astrophysics Data System (ADS)

    Haines-Stiles, G.; Rabello-Soares, C.

    2006-05-01

    The International Geophysical Year of 1957-1958 revolutionized Earth and Space science. But the "official" IGY films, sponsored by the National Academies, debuted almost two years after field research was completed. For 2007-2009, communications revolutions such as the Internet and satellite voice and video enable contemporaneous public engagement via media, at science centers, and at home, and opportunities at all levels of formal education impossible in 1957. Real-time communication to both Poles, video podcasts from explorers of ice sheets and near-Earth space, and inquiry-based educational modules for schools and community centers, provide practical but inspirational ways to engage, inform and inspire millions across Earth in new ways, supporting the outreach goals of all four International Science Years. Examples of live video connections to NSF's Amundsen-Scott South Pole Station, and to an Alaskan sounding rocket range, together with evaluation data on a rock-concert inspired "tour" of science centers, schools and community venues by NASA researchers and engineers, provide possible models for the upcoming ISY's. A calendar of polar field campaigns, NASA launches and events (such as arrival of the Lunar Reconnaissance Orbiter at the Moon in Fall 2008, and the landing of Mars Phoenix in Spring 2008), and earth and space science related anniversaries - such as the launch of Explorer 1 on January 31, 1958 - offer a framework for integrating outreach, engagement and education strategies common to all four Science Years.

  15. Cloning and characterization of a SnRK2 gene from Jatropha curcas L.

    PubMed

    Chun, J; Li, F-S; Ma, Y; Wang, S-H; Chen, F

    2014-01-01

    Although the SnRK2 class of Ser/Thr protein kinases is critical for signal transduction and abiotic stress resistance in plants, there have been no studies to examine SnRK2 in Jatropha curcas L. In the present study, JcSnRK2 was cloned from J. curcas using the rapid amplification of cDNA end technique and characterized. The JcSnRK2 genomic sequence is 2578 base pairs (bp), includes 10 exons and 9 introns, and the 1017-bp open reading frame encodes 338 amino acids. JcSnRK2 was transcribed in all examined tissues, with the highest transcription rate observed in the roots, followed by the leaves and stalks; the lowest rate was observed in flowers and seeds. JcSnRK2 expression increased following abscisic acid treatment, salinity, and drought stress. During a 48-h stress period, the expression of JcSnRK2 showed 2 peaks and periodic up- and downregulation. JcSnRK2 was rapidly activated within 1 h under salt and drought stress, but not under cold stress. Because of the gene sequence and expression similarity of JcSnRK2 to AtSnRK2.8, primarily in the roots, an eukaryotic expression vector containing the JcSnRK2 gene (pBI121-JcSnRK2) was constructed and introduced to the Arabidopsis AtSnRK2.8 mutant snf2.8. JcSnRK2-overexpressing plants exhibited higher salt and drought tolerance, further demonstrating the function of JcSnRK2 in the osmotic stress response. J. curcas is highly resistant to extreme salt and drought conditions and JcSnRK2 was found to be activated under these conditions. Thus, JcSnRK2 is potential candidate for improving crop tolerance to salt and drought stress. PMID:25526217

  16. Egg yolk IgY: protection against rotavirus induced diarrhea and modulatory effect on the systemic and mucosal antibody responses in newborn calves.

    PubMed

    Vega, C; Bok, M; Chacana, P; Saif, L; Fernandez, F; Parreño, V

    2011-08-15

    Bovine rotavirus (BRV) is an important cause of diarrhea in newborn calves. Local passive immunity is the most efficient protective strategy to control the disease. IgY technology (the use of chicken egg yolk immunoglobulins) is an economic and practical alternative to prevent BRV diarrhea in dairy calves. The aim of this study was to evaluate the protection and immunomodulation induced by the oral administration of egg yolk enriched in BRV specific IgY to experimentally BRV infected calves. All calves in groups Gp 1, 2 and 3 received control colostrum (CC; BRV virus neutralization Ab titer - VN=65,536; ELISA BRV IgG(1)=16,384) prior to gut closure. After gut closure, calves received milk supplemented with 6% BRV-immune egg yolk [(Gp 1) VN=2048; ELISA IgY Ab titer=4096] or non-immune control egg yolk [(Gp 2) VN<4; ELISA IgY Ab titer<4] twice a day, for 14 days. Calves receiving CC only or colostrum deprived calves (CD) fed antibody (Ab) free milk served as controls (Gp 3 and 4, respectively). Calves were inoculated with 10(5.85)focus forming units (FFU) of virulent BRV IND at 2 days of age. Control calves (Gp 3 and 4) and calves fed control IgY (Gp 2) were infected and developed severe diarrhea. Around 80% calves in Gp 1 (IgY 4096) were infected, but they showed 80% (4/5) protection against BRV diarrhea. Bovine RV-specific IgY Ab were detected in the feces of calves in Gp 1, indicating that avian antibodies (Abs) remained intact after passage through the gastrointestinal tract. At post infection day 21, the duodenum was the major site of BRV specific antibody secreting cells (ASC) in all experimental groups. Mucosal ASC responses of all isotypes were significantly higher in the IgY treated groups, independently of the specificity of the treatment, indicating that egg yolk components modulated the immune response against BRV infection at the mucosal level. These results indicate that supplementing newborn calves' diets for the first 14 days of life with egg yolk

  17. Heterologous Overexpression of Poplar SnRK2 Genes Enhanced Salt Stress Tolerance in Arabidopsis thaliana

    PubMed Central

    Song, Xueqing; Yu, Xiang; Hori, Chiaki; Demura, Taku; Ohtani, Misato; Zhuge, Qiang

    2016-01-01

    Subfamily 2 of SNF1-related protein kinase (SnRK2) plays important roles in plant abiotic stress responses as a global positive regulator of abscisic acid signaling. In the genome of the model tree Populus trichocarpa, 12 SnRK2 genes have been identified, and some are upregulated by abiotic stresses. In this study, we heterologously overexpressed the PtSnRK2 genes in Arabidopsis thaliana and found that overexpression of PtSnRK2.5 and PtSnRK2.7 genes enhanced stress tolerance. In the PtSnRK2.5 and PtSnRK2.7 overexpressors, chlorophyll content, and root elongation were maintained under salt stress conditions, leading to higher survival rates under salt stress compared with those in the wild type. Transcriptomic analysis revealed that PtSnRK2.7 overexpression affected stress-related metabolic genes, including lipid metabolism and flavonoid metabolism, even under normal growth conditions. However, the stress response genes reported to be upregulated in Arabidopsis SRK2C/SnRK2.6 and wheat SnRK2.8 overexpressors were not changed by PtSnRK2.7 overexpression. Furthermore, PtSnRK2.7 overexpression widely and largely influenced the transcriptome in response to salt stress; genes related to transport activity, including anion transport-related genes, were characteristically upregulated, and a variety of metabolic genes were specifically downregulated. We also found that the salt stress response genes were greatly upregulated in the PtSnRK2.7 overexpressor. Taken together, poplar subclass 2 PtSnRK2 genes can modulate salt stress tolerance in Arabidopsis, through the activation of cellular signaling pathways in a different manner from that by herbal subclass 2 SnRK2 genes. PMID:27242819

  18. Do r/K reproductive strategies apply to human differences?

    PubMed

    Rushton, J P

    1988-01-01

    This article discusses the r/K theory of Social Biology and how it relates to humans. The symbols r and K originate in the mathematics of population biology and refer to 2 ends of a continuum in which a compensatory exchange occurs between gamete production (the r-strategy) and longevity (the K-strategy). Both across and within species, r and K strategists differ in a suite of correlated characteristics. Humans are the most K of all. K's supposedly have a longer gestation period, a higher birthweight, a more delayed sexual maturation, a lower sex drive, and a longer life. Studies providing evidence for the expected covariation among K attributes are presented. Additional evidence for r/K theory comes from the comparison of human population known to differ in gamete production. The pattern of racial differences observed to occur in sexual behavior has also been found to exist on numerous other indices of K. For instance, there are racial differences in brain size, intelligence, and maturation rate, among others. The findings suggest that, on the average, Mongoloids are more K than Caucasoids, who in turn, are more K than Negroids. Recently conducted studies have extended the data in favor of r/K theory, and further research is currently underway, including whether r/K attributes underlie individual and social class differences in health and longevity. PMID:3241997

  19. Genome-wide analysis of SnRK gene family in Brachypodium distachyon and functional characterization of BdSnRK2.9.

    PubMed

    Wang, Lianzhe; Hu, Wei; Sun, Jiutong; Liang, Xiaoyu; Yang, Xiaoyue; Wei, Shuya; Wang, Xiatian; Zhou, Yi; Xiao, Qiang; Yang, Guangxiao; He, Guangyuan

    2015-08-01

    The sucrose non-fermenting 1 (SNF1)-related protein kinases (SnRKs) play key roles in plant signaling pathways including responses to biotic and abiotic stresses. Although SnRKs have been systematically studied in Arabidopsis and rice, there is no information concerning SnRKs in the new Poaceae model plant Brachypodium distachyon. In the present study, a total of 44 BdSnRKs were identified and classified into three subfamilies, including three members of BdSnRK1, 10 of BdSnRK2 and 31 of BdSnRK3 (CIPK) subfamilies. Phylogenetic reconstruction, chromosome distribution and synteny analyses suggested that BdSnRK family had been established before the dicot-monocot lineage parted, and had experienced rapid expansion during the process of plant evolution since then. Expression analysis of the BdSnRK2 subfamily showed that the majority of them could respond to abiotic stress and related signal molecules treatments. Protein-protein interaction and co-expression analyses of BdSnRK2s network showed that SnRK2s might be involved in biological pathway different from that of dicot model plant Arabidopsis. Expression of BdSnRK2.9 in tobacco resulted in increased tolerance to drought and salt stresses through activation of NtABF2. Taken together, comprehensive analyses of BdSnRKs would provide a basis for understanding of evolution and function of BdSnRK family. PMID:26089150

  20. PLGA nanoparticle formulation of RK-33: an RNA helicase inhibitor against DDX3

    PubMed Central

    Bol, Guus Martinus; Khan, Raheela; van Voss, Marise Rosa Heerma; Tantravedi, Saritha; Korz, Dorian

    2016-01-01

    Background The DDX3 helicase inhibitor RK-33 is a newly developed anticancer agent that showed promising results in preclinical research (Bol et al. EMBO Mol Med, 7(5):648–649, 2015). However, due to the physicochemical and pharmacological characteristics of RK-33, we initiated development of alternative formulations of RK-33 by preparing sustained release nanoparticles that can be administered intravenously. Methods In this study, RK-33 was encapsulated in poly(lactic-co-glycolic acid) (PLGA), one of the most well-developed biodegradable polymers, using the emulsion solvent evaporation method. Results Hydrodynamic diameter of RK-33-PLGA nanoparticles was about 245 nm with a negative charge, and RK-33-PLGA nanoparticles had a payload of 1.4 % RK-33. RK-33 was released from the PLGA nanoparticles over 7 days (90 ± 5.7 % released by day 7) and exhibited cytotoxicity to human breast carcinoma MCF-7 cells in a time-dependent manner. Moreover, RK-33-PLGA nanoparticles were well tolerated, and systemic retention of RK-33 was markedly improved in normal mice. Conclusions PLGA nanoparticles have a potential as a parenteral formulation of RK-33. PMID:26330329

  1. Anti-Pseudomonas aeruginosa IgY Antibodies Induce Specific Bacterial Aggregation and Internalization in Human Polymorphonuclear Neutrophils

    PubMed Central

    Thomsen, K.; Christophersen, L.; Bjarnsholt, T.; Jensen, P. Ø.; Moser, C.

    2015-01-01

    Polymorphonuclear neutrophils (PMNs) are essential cellular constituents in the innate host response, and their recruitment to the lungs and subsequent ubiquitous phagocytosis controls primary respiratory infection. Cystic fibrosis pulmonary disease is characterized by progressive pulmonary decline governed by a persistent, exaggerated inflammatory response dominated by PMNs. The principal contributor is chronic Pseudomonas aeruginosa biofilm infection, which attracts and activates PMNs and thereby is responsible for the continuing inflammation. Strategies to prevent initial airway colonization with P. aeruginosa by augmenting the phagocytic competence of PMNs may postpone the deteriorating chronic biofilm infection. Anti-P. aeruginosa IgY antibodies significantly increase the PMN-mediated respiratory burst and subsequent bacterial killing of P. aeruginosa in vitro. The mode of action is attributed to IgY-facilitated formation of immobilized bacteria in aggregates, as visualized by fluorescence microscopy and the induction of increased bacterial hydrophobicity. Thus, the present study demonstrates that avian egg yolk immunoglobulins (IgY) targeting P. aeruginosa modify bacterial fitness, which enhances bacterial killing by PMN-mediated phagocytosis and thereby may facilitate a rapid bacterial clearance in airways of people with cystic fibrosis. PMID:25895968

  2. Anti-Interleukin-1 Beta/Tumor Necrosis Factor-Alpha IgY Antibodies Reduce Pathological Allergic Responses in Guinea Pigs with Allergic Rhinitis

    PubMed Central

    Wei-xu, Hu; Wen-yun, Zhou; Xi-ling, Zhu; Zhu, Wen; Li-hua, Wu; Xiao-mu, Wu; Hui-ping, Wei; Wen-ding, Wang; Dan, He; Qin, Xiang

    2016-01-01

    This study aims to determine whether the combined blockade of IL-1β and TNF-α can alleviate the pathological allergic inflammatory reaction in the nasal mucosa and lung tissues in allergic rhinitis (AR) guinea pigs. Healthy guinea pigs treated with saline were used as the healthy controls. The AR guinea pigs were randomly divided into (1) the AR model group treated with intranasal saline; (2) the 0.1% nonspecific IgY treatment group; (3) the 0.1% anti-TNF-α IgY treatment group; (4) the 0.1% anti-IL-1β IgY treatment group; (5) the 0.1% combined anti-IL-1β and TNF-α IgY treatment group; and (6) the fluticasone propionate treatment group. The inflammatory cells were evaluated using Wright's staining. Histopathology was examined using hematoxylin-eosin staining. The results showed that the number of eosinophils was significantly decreased in the peripheral blood, nasal lavage fluid, and bronchoalveolar lavage fluid (P < 0.05), and eosinophil, neutrophil, and lymphocyte infiltration and edema were significantly reduced or absent in the nasal mucosa and lung tissues (P < 0.05) in the combined 0.1% anti-IL-1β- and TNF-α IgY-treated guinea pigs. The data suggest that topical blockade of IL-1β and TNF-α could reduce pathological allergic inflammation in the nasal mucosa and lung tissues in AR guinea pigs. PMID:27046957

  3. Production of specific IgY Helicobacter pylori recombinant OipA protein and assessment of its inhibitory effects towards attachment of H. pylori to AGS cell line

    PubMed Central

    Borhani, Katayoun; Behmanesh, Mehrdad; Khoramabadi, Nima

    2015-01-01

    Purpose The common triple therapy for Helicobacter pylori is challenged by the increasing cases of antibiotic resistant infections, raising the need to explore alternative therapies. Oral administration of egg yolk immunoglobulin Y (IgY) has been previously reported as a means of passive immunization therapy for H. pylori infections. In this work, we investigated the inhibitory effect of IgY on the attachment of H. pylori to AGS cell line. Materials and Methods Recombinant OipA was prepared. Hens were immunized with recombinant protein three times. IgY was purified from egg yolks of immunized hens using polyethylene glycol precipitation method. The inhibitory effect of the specific immunoglobulin was evaluated in AGS cell line infected with H. pylori. Results The presence of recombinant OipA (30 kD) was confirmed via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Immunization of hens was confirmed using enzyme-linked immunosorbent assay. The purified IgY from egg yolks were assessed using SDS-PAGE and confirmed by western blot. Conclusion The results showed that IgY-OipA had inhibitory effect on attachment of H. pylori to AGS cell line and may be utilized as a therapeutic or prophylaxis material. PMID:26273576

  4. Chicken egg yolk anti-asialoGM1 immunoglobulin (IgY): an inexpensive glycohistochemical probe for localization of T-antigen in human colorectal adenocarcinomas.

    PubMed

    Sriram, V; Jebaraj, C E; Yogeeswaran, G

    1999-07-01

    A egg yolk polyclonal IgY has been prepared by immunization of white leghorn chickens with small unilamellar liposomal asialoGM1. The newly prepared anti-asialoGM1 IgY has been characterized to be specific toward the terminal carbohydrate moiety of asialoGM1, and has no cross reactivity to its sialylated counterpart (ganglioside, GM1) as evidenced by immunochromatographic studies. General glycohistochemical methods along with antigen specific lectin and immunohistochemical staining using anti-asialoGM1 IgY were used to study the expression of Thomsen-Friedenreich (T-) disaccharide antigen in human colorectal adenocarcinoma tissues. The expression of T-antigen in colon cancer tissue was detected by two T-disaccharide specific probes, chicken anti-T-yolk antibody (IgY) and Artocarpus integrifolia lectin (AIL) and was found to be more pronounced in both the secreted mucin as well as the cytoplasmic mucin deposits. These immunochemical detection methods for T-antigen showed a weaker correlation with other glycostaining methods using, alcian-blue/periodic acid-Schiff (AB-PAS) and high iron diamine (HID). However, a general enzymatic staining for galactose and galactosamine containing glycoconjugates, by galactose oxidase-Schiff method, showed a good correlation with T-antigen detection. While the T-beta specific anti-asialoGM1 could localize T-antigen in 11 of 13 (84%) human colorectal adenocarcinoma tissue sections tested, the T-alpha specific AIL could localize the T-antigen in only 6 of the tissues (46%). These observations confirm previously reported findings, of the prevalence of T-beta conformation in colon cancer, that binds significantly more with the anti-asialoGM1 IgY than with the T-alpha specific AIL. Hence, both anti-T IgY and the AIL immunohistochemical probes may have useful diagnostic value because of the ease of preparation and cost effectiveness, but the T-beta specific anti-asialoGM1 probe (IgY) would have a better prognostic value in colon

  5. Differential Activation of the Wheat SnRK2 Family by Abiotic Stresses

    PubMed Central

    Zhang, Hongying; Li, Weiyu; Mao, Xinguo; Jing, Ruilian; Jia, Hongfang

    2016-01-01

    Plant responses to stress occur via abscisic acid (ABA) dependent or independent pathways. Sucrose non-fermenting1-related protein kinase 2 (SnRK2) play a key role in plant stress signal transduction pathways. It is known that some SnRK2 members are positive regulators of ABA signal transduction through interaction with group A type 2C protein phosphatases (PP2Cs). Here, 10 SnRK2s were isolated from wheat. Based on phylogenetic analysis using kinase domains or the C-terminus, the 10 SnRK2s were divided into three subclasses. Expression pattern analysis revealed that all TaSnRK2s were involved in the responses to PEG, NaCl, and cold stress. TaSnRK2s in subclass III were strongly induced by ABA. Subclass II TaSnRK2s responded weakly to ABA, whereas TaSnRK2s in subclass I were not activated by ABA treatment. Motif scanning in the C-terminus indicated that motifs 4 and 5 in the C-terminus were unique to subclass III. We further demonstrate the physical and functional interaction between TaSnRK2s and a typical group A PP2C (TaABI1) using Y2H and BiFC assays. The results showed that TaABI1 interacted physically with subclass III TaSnRK2s, while having no interaction with subclasses I and II TaSnRK2s. Together, these findings indicated that subclass III TaSnRK2s were involved in ABA regulated stress responses, whereas subclasses I and II TaSnRK2s responded to various abiotic stressors in an ABA-independent manner. PMID:27066054

  6. From the IGY to the IHY: A Changing View of the Van Allen Radiation Belts

    NASA Astrophysics Data System (ADS)

    Hudson, M. K.

    2006-12-01

    with the choice of solar maximum for the IGY. Still, much remains to be done in terms of predictive capability. The need for such capability gave rise to establishing the National Space Weather Program a decade ago, while the fundamental science questions of solar-terrestrial connectivity remain drivers for NASA's Living With a Star program and other focused projects such as the NSF-sponsored Center for Integrated Space Weather Modeling. As measurements of emerging solar active regions have greatly improved, soon to be given a STERIO view, computer modeling capability has developed beyond the wildest expectations of Professor Van Allen and colleagues who launched the first Geiger counter into space on a satellite with no data storage device (once called a tape recorder). Results from recent simulations attempting both to describe particle acceleration processes quantitatively and predict impact on the near Earth space environment will be presented.

  7. IgY14 and SuperMix immunoaffinity separations coupled with liquid chromatography-mass spectrometry for human plasma proteomic biomarker discovery

    SciTech Connect

    Shi, Tujin; Zhou, Jianying; Gritsenko, Marina A.; Hossain, Mahmud; Camp, David G.; Smith, Richard D.; Qian, Weijun

    2012-02-01

    Interest in the application of advanced proteomics technologies to human blood plasma- or serum-based clinical samples for the purpose of discovering disease biomarkers continues to grow; however, the enormous dynamic range of protein concentrations in these types of samples (often >10 orders of magnitude) represents a significant analytical challenge, particularly for detecting low-abundance candidate biomarkers. In response, immunoaffinity separation methods for depleting multiple high- and moderate-abundance proteins have become key tools for enriching low-abundance proteins and enhancing detection of these proteins in plasma proteomics. Herein, we describe IgY14 and tandem IgY14-Supermix separation methods for removing 14 high-abundance and up to 60 moderate-abundance proteins, respectively, from human blood plasma and highlight their utility when combined with liquid chromatography-tandem mass spectrometry for interrogating the human plasma proteome.

  8. Health Information in Turkish (Türkçe): MedlinePlus

    MedlinePlus

    ... Videos & Tools You Are Here: Home → Multiple Languages → Turkish (Türkçe) URL of this page: https://www.nlm.nih.gov/medlineplus/languages/turkish.html Health Information in Turkish (Türkçe) To use ...

  9. Development and evaluation of IgY ImmunoCapture PCR ELISA for detection of Staphylococcus aureus enterotoxin A devoid of protein A interference.

    PubMed

    Reddy, Prakash; Ramlal, Shylaja; Sripathy, Murali Harishchandra; Batra, Harsh Vardhan

    2014-06-01

    In the present study, a sensitive and specific IgY mediated ImmunoCapture-PCR-ELISA (IC-PCR-ELISA) was developed for the detection of staphylococcal enterotoxin A (SEA) from culture supernatants and suspected contaminated samples. Due to the virtue of avian immunoglobulins (IgY) to have the least affinity towards staphylococcal protein A (SpA) responsible for false positives, we employed anti-SEA IgY for capture of SEA toxin and revealed with SEA specific rabbit antibodies conjugated to a 524bp DNA marker. Biotin-11-dUTP was incorporated during PCR amplification and post PCR analysis was performed by PCR-ELISA. Unlike IgG immunocapture, IgY mediated immunocapture of SEA was free from false positives due to protein A. The developed assay was specific to SEA except for minor cross reactivity with staphylococcal enterotoxin E (SEE). Several raw milk samples were evaluated for the presence of SEA with and without enrichment. Three samples were found to be positive for SEA after enrichment for 8h. Though IC-PCR-ELISA for SEA showed 100% correlation with PCR analysis for sea gene, the assay was unique in terms of sensitivity of detecting ~10pg/ml of SEA toxin from spiked milk samples. Result of IC-PCR-ELISA was further confirmed by conventional methods of isolation and characterization. The presented method can be very useful for rapid analysis of milk samples for SEA contamination and can be further extended for detection of multiple SE's in different wells of same PCR plate using common DNA substrate. PMID:24941881

  10. Mouse-Specific Tandem IgY7-SuperMix Immunoaffinity Separations for Improved LC-MS/MS Coverage of the Plasma Proteome

    SciTech Connect

    Zhou, Jianying; Petritis, Brianne O.; Petritis, Konstantinos; Norbeck, Angela D.; Weitz, Karl K.; Moore, Ronald J.; Camp, David G.; Kulkarni, Rohit N.; Smith, Richard D.; Qian, Weijun

    2009-09-01

    We report on a customized mouse specific SuperMix immunoaffinity column and strategy for separating low abundance proteins from high and moderate abundance proteins in mouse plasma. When applied in tandem with a mouse IgY7 column that removes the seven most abundant proteins in blood, the SuperMix column captures >100 additional moderate abundance proteins, thus allowing significant enrichment of low abundance proteins in the flow-through fraction. A side-by-side comparison of results obtained from 2D-LC-MS/MS analyses of flow-through samples from IgY7 and SuperMix columns revealed a nearly two-fold improvement in the overall proteome coverage. Detection of low abundance proteins was also enhanced, as evidenced by a more than two-fold increase in the coverage of cytokines, growth factors, and other low abundance proteins. Moreover, the tandem separations are automated, reproducible, and allow effective identification of protein abundance differences from LC-MS/MS analyses. Considering the overall reproducibility and increased sensitivity using the IgY7-SuperMix separation system, we anticipate broad applications of this strategy for biomarker discovery using mouse models.

  11. Mouse-Specific Tandem IgY7-SuperMix Immunoaffinity Separations for Improved LC-MS/MS Coverage of the Plasma Proteome

    PubMed Central

    Zhou, Jian-Ying; Petritis, Brianne O.; Petritis, Konstantinos; Norbeck, Angela D.; Weitz, Karl K.; Moore, Ronald J.; Camp, David G.; Kulkarni, Rohit N.; Smith, Richard D.; Qian, Wei-Jun

    2009-01-01

    We report on a mouse specific SuperMix immunoaffinity separation system for separating low abundance proteins from high and moderate abundance proteins in mouse plasma. When applied in tandem with a mouse IgY7 column that removes the seven most abundant proteins in plasma, the SuperMix column captures more than 100 additional moderate abundance proteins, thus allowing significant enrichment of low abundance proteins in the flow-through fraction. A side-by-side comparison of results obtained from 2D-LC-MS/MS analyses of flow-through samples from IgY7 and SuperMix columns revealed a nearly two-fold improvement in the overall proteome coverage. Detection of low abundance proteins was also enhanced, as evidenced by a more than two-fold increase in the coverage of cytokines, growth factors, and other low abundance proteins. Moreover, the tandem separations are automated, reproducible, and allow effective identification of protein abundance differences from LC-MS/MS analyses. Considering the overall reproducibility and increased sensitivity using the IgY7-SuperMix separation system, we anticipate broad applications of this strategy for biomarker discovery using mouse models. PMID:19722698

  12. Preparation of immunoglobulin Y (IgY) against lipopolysaccharide using gel chromatography from the yolks of eggs laid by immunized hens.

    PubMed

    Ma, Siyuan; Zhang, Yaping

    2010-10-01

    The objective is to prevent and treat injuries caused by lipopolysaccharide (LPS) from gram negative bacteria in animals and humans, we produced antibodies against LPS from egg yolk. LPS from E. coli (O111:B4) mixed with Freund's Adjuvant was used as the immunogen to immunize Roman hens. Immunized eggs were collected, and immunoglobulin Y (IgY) was purified using a water solution, salt precipitation and gel chromatography. The molecular weight and purity were determined by SDS-PAGE, the antibody titer by noncompetitive enzyme-linked immunosorbent assay (ELISA) and antibody activity against LPS by the mortality of mice intraperitoneally injected with LPS or LPS-IgY solutions. IgY against LPS showed two protein bands at 68 and 26 kDa on the gel; the antibody titer was almost 1:25,600. After incubation with LPS, IgY decreased the mortality of mice challenged with LPS. This study provided an efficient way to produce high-titer egg yolk antibodies, which could attenuate lethal effects of LPS, by immunizing hens. Furthermore, the LPS antibody was purified well using a water solution, salting-out and gel chromatography. PMID:20798981

  13. A transfectant RK13 cell line permissive to classical caprine scrapie prion propagation.

    PubMed

    Dassanayake, Rohana P; Zhuang, Dongyue; Truscott, Thomas C; Madsen-Bouterse, Sally A; O'Rourke, Katherine I; Schneider, David A

    2016-03-01

    To assess scrapie infectivity associated with caprine-origin tissues, bioassay can be performed using kids, lambs or transgenic mice expressing caprine or ovine prion (PRNP) alleles, but the incubation periods are fairly long. Although several classical ovine scrapie prion permissive cell lines with the ability to detect brain-derived scrapie prion have been available, no classical caprine scrapie permissive cell line is currently available. Therefore, the aims of this study were to generate a rabbit kidney epithelial cell line (RK13) stably expressing caprine wild-type PRNP (cpRK13) and then to assess permissiveness of cpRK13 cells to classical caprine scrapie prion propagation. The cpRK13 and plasmid control RK13 (pcRK13) cells were incubated with brain-derived classical caprine scrapie inocula prepared from goats or ovinized transgenic mice (Tg338, express ovine VRQ allele) infected with caprine scrapie. Significant PrP(Sc) accumulation, which is indicative of scrapie prion propagation, was detected by TSE ELISA and immunohistochemistry in cpRK13 cells inoculated with classical caprine scrapie inocula. Western blot analysis revealed the typical proteinase K-resistant 3 PrP(res) isoforms in the caprine scrapie prion inoculated cpRK13 cell lysate. Importantly, PrP(Sc) accumulation was not detected in similarly inoculated pcRK13 cells, whether by TSE ELISA, immunohistochemistry, or western blot. These findings suggest that caprine scrapie prions can be propagated in cpRK13 cells, thus this cell line may be a useful tool for the assessment of classical caprine prions in the brain tissues of goats. PMID:27216989

  14. Simultaneous explanation of the RK and R (D (*)) puzzles

    NASA Astrophysics Data System (ADS)

    Bhattacharya, Bhubanjyoti; Datta, Alakabha; London, David; Shivashankara, Shanmuka

    2015-03-01

    At present, there are several hints of lepton flavor non-universality. The LHCb Collaboration has measured RK ≡ B (B+ →K+μ+μ-) / B (B+ →K+e+e-), and the BaBar Collaboration has measured R (D (*)) ≡ B (B bar →D (*) +τ-νbarτ) / B (B bar →D (*) +ℓ-νbarℓ) (ℓ = e , μ). In all cases, the experimental results differ from the standard model predictions by 2- 3 σ. Recently, an explanation of the RK puzzle was proposed in which new physics (NP) generates a neutral-current operator involving only third-generation particles. Now, assuming the scale of NP is much larger than the weak scale, this NP operator must be made invariant under the full SU (3)C × SU (2)L × U(1)Y gauge group. In this Letter, we note that, when this is done, a new charged-current operator can appear, and this can explain the R (D (*)) puzzle. A more precise measurement of the double ratio R (D) / R (D*) can rule out this model.

  15. Detection of Clonorchis sinensis Circulating Antigen in Sera from Chinese Patients by Immunomagnetic Bead ELISA Based on IgY

    PubMed Central

    Nie, Ge; Wang, Ting; Lu, Shengjun; Liu, Wenqi; Li, Yonglong; Lei, Jiahui

    2014-01-01

    Background Clonorchiasis, caused by Clonorchis sinensis, is widely distributed in Southeast Asia including China. Clonorchiasis is included in control programs of neglected tropical diseases by World Health Organization (WHO) because it is one of the major health problems in most endemic areas. Diagnosis of clonorchiasis plays a key role in the control programs. However, so far, there is no satisfactory method for clonorchiasis because of low sensitivity, poor practicality and high false positivity of available diagnostic tools. Methodology/Principal Findings We developed an immunomagnetic bead enzyme-linked immunosorbent assay (ELISA) based on IgY (egg yolk immunoglobulin) against cysteine proteinase of C. sinensis for detection of circulating antigen in serum samples of patients infected with C. sinensis. The polyclonal IgY, coated with magnetic beads, was used as a capture antibody and a monoclonal IgG labeled with horseradish peroxidase as a detection antibody in the IgY-based immunomagnetic bead ELISA system (IgY-IMB-ELISA). The results showed that the sensitivity of IgY-IMB-ELISA was 93.3% (14 of 15) in cases of heavy infection (5000 to 9999 eggs per gram feces, i.e, EPG 5000–9999), 86.7% (13 of 15) in cases of moderate infection (EPG 1000–4999) and 75.0% (9 of 12) in cases of light infection (EPG <1000) of clonorchiasis. Together 36 of total 42 (85.7%) serum samples of human clonorchiasis gave a positive reaction. There was a significant correlation between ELISA optical density and egg counts (EPG) with a correlation coefficient of 0.83 in total 42 patients. There were no positive results in patients with trichinosis (n = 10) or cysticercosis (n = 10). Cross-reactivity was 6.7% (2 of 30) with schistosomiasis japonica and 10.0% (3 of 30) with paragonimiasis, respectively. No positive reaction was found in 20 healthy persons. Conclusions Our findings suggest that IgY-IMB-ELISA appears to be a sensitive and specific assay for detection of

  16. The IGY and the Satellite Race: A Reconsideration of a Cold War Crisis that Never Should Have Been

    NASA Astrophysics Data System (ADS)

    Launius, R. D.

    2006-05-01

    In October 1957 the Soviet Union launched the first Earth-circling artificial satellite and the crisis that resulted led to numerous actions in the United States aimed at "remediating" a Cold War crisis. This included the establishment of a separate civilian space agency charged with the conduct of an official program of scientific and technological space exploration, consolidation of Department of Defense space activities, the passage of the National Defense Education Act, the creation of a Presidential Science Advisor, and a host of lesser actions. The politics of these changes is fascinating, and has been interpreted as an appropriate political response to a unique crisis situation. Interest groups, all for differing reasons, prodded national leaders to undertake large-scale efforts, something the president thought unnecessarily expensive and once set in place impossible to dismantle. But was the Sputnik crisis truly a crisis in any real sense? Was it made into one by interest groups who used it for their own ends? This paper will trace briefly some of the major themes associated with the IGY and Sputnik and describe the political construction of the crisis as it emerged in 1957- 1958. It will also discuss something about the transformation of federal science and technology that took place in the aftermath of the "crisis" and how it set in train a series of processes and policies that did not unravel until the end of the Cold War.

  17. Quantitative phosphoproteomics of protein kinase SnRK1 regulated protein phosphorylation in Arabidopsis under submergence.

    PubMed

    Cho, Hsing-Yi; Wen, Tuan-Nan; Wang, Ying-Tsui; Shih, Ming-Che

    2016-04-01

    SNF1 RELATED PROTEIN KINASE 1 (SnRK1) is proposed to be a central integrator of the plant stress and energy starvation signalling pathways. We observed that the Arabidopsis SnRK1.1 dominant negative mutant (SnRK1.1 (K48M) ) had lower tolerance to submergence than the wild type, suggesting that SnRK1.1-dependent phosphorylation of target proteins is important in signalling pathways triggered by submergence. We conducted quantitative phosphoproteomics and found that the phosphorylation levels of 57 proteins increased and the levels of 27 proteins decreased in Col-0 within 0.5-3h of submergence. Among the 57 proteins with increased phosphorylation in Col-0, 38 did not show increased phosphorylation levels in SnRK1.1 (K48M) under submergence. These proteins are involved mainly in sugar and protein synthesis. In particular, the phosphorylation of MPK6, which is involved in regulating ROS responses under abiotic stresses, was disrupted in the SnRK1.1 (K48M) mutant. In addition, PTP1, a negative regulator of MPK6 activity that directly dephosphorylates MPK6, was also regulated by SnRK1.1. We also showed that energy conservation was disrupted in SnRK1.1 (K48M) , mpk6, and PTP1 (S7AS8A) under submergence. These results reveal insights into the function of SnRK1 and the downstream signalling factors related to submergence. PMID:27029354

  18. Quantitative phosphoproteomics of protein kinase SnRK1 regulated protein phosphorylation in Arabidopsis under submergence

    PubMed Central

    Cho, Hsing-Yi; Wen, Tuan-Nan; Wang, Ying-Tsui; Shih, Ming-Che

    2016-01-01

    SNF1 RELATED PROTEIN KINASE 1 (SnRK1) is proposed to be a central integrator of the plant stress and energy starvation signalling pathways. We observed that the Arabidopsis SnRK1.1 dominant negative mutant (SnRK1.1 K48M) had lower tolerance to submergence than the wild type, suggesting that SnRK1.1-dependent phosphorylation of target proteins is important in signalling pathways triggered by submergence. We conducted quantitative phosphoproteomics and found that the phosphorylation levels of 57 proteins increased and the levels of 27 proteins decreased in Col-0 within 0.5–3h of submergence. Among the 57 proteins with increased phosphorylation in Col-0, 38 did not show increased phosphorylation levels in SnRK1.1 K48M under submergence. These proteins are involved mainly in sugar and protein synthesis. In particular, the phosphorylation of MPK6, which is involved in regulating ROS responses under abiotic stresses, was disrupted in the SnRK1.1 K48M mutant. In addition, PTP1, a negative regulator of MPK6 activity that directly dephosphorylates MPK6, was also regulated by SnRK1.1. We also showed that energy conservation was disrupted in SnRK1.1 K48M, mpk6, and PTP1 S7AS8A under submergence. These results reveal insights into the function of SnRK1 and the downstream signalling factors related to submergence. PMID:27029354

  19. Molecular Mimicry Regulates ABA Signaling by SnRK2 Kinases and PP2C Phosphatases

    SciTech Connect

    Soon, Fen-Fen; Ng, Ley-Moy; Zhou, X. Edward; West, Graham M.; Kovach, Amanda; Tan, M.H. Eileen; Suino-Powell, Kelly M.; He, Yuanzheng; Xu, Yong; Chalmers, Michael J.; Brunzelle, Joseph S.; Zhang, Huiming; Yang, Huaiyu; Jiang, Hualiang; Li, Jun; Yong, Eu-Leong; Cutler, Sean; Zhu, Jian-Kang; Griffin, Patrick R.; Melcher, Karsten; Xu, H. Eric

    2014-10-02

    Abscisic acid (ABA) is an essential hormone for plants to survive environmental stresses. At the center of the ABA signaling network is a subfamily of type 2C protein phosphatases (PP2Cs), which form exclusive interactions with ABA receptors and subfamily 2 Snfl-related kinase (SnRK2s). Here, we report a SnRK2-PP2C complex structure, which reveals marked similarity in PP2C recognition by SnRK2 and ABA receptors. In the complex, the kinase activation loop docks into the active site of PP2C, while the conserved ABA-sensing tryptophan of PP2C inserts into the kinase catalytic cleft, thus mimicking receptor-PP2C interactions. These structural results provide a simple mechanism that directly couples ABA binding to SnRK2 kinase activation and highlight a new paradigm of kinase-phosphatase regulation through mutual packing of their catalytic sites.

  20. Detection of Shiga toxin-producing Escherichia coli by sandwich enzyme-linked immunosorbent assay using chicken egg yolk IgY antibodies

    PubMed Central

    Parma, Y. R.; Chacana, P. A.; Lucchesi, P. M. A.; Rogé, A.; Granobles Velandia, C. V.; Krüger, A.; Parma, A. E.; Fernández-Miyakawa, M. E.

    2012-01-01

    Enterohemorrhagic Escherichia coli (EHEC), a subset of Shiga toxin producing E. coli (STEC) is associated with a spectrum of diseases that includes diarrhea, hemorrhagic colitis and a life-threatening hemolytic-uremic syndrome (HUS). Regardless of serotype, Shiga toxins (Stx1 and/or Stx2) are uniformly expressed by all EHEC, and so exploitable targets for laboratory diagnosis of these pathogens. In this study, a sandwich ELISA for determination of Shiga toxin (Stx) was developed using anti-Stx2B subunit antibodies and its performance was compared with that of the Vero cell assay and a commercial immunoassay kit. Chicken IgY was used as capture antibody and a HRP-conjugated rabbit IgG as the detection antibody. The anti-Stx2B IgY was harvested from eggs laid by hens immunized with a recombinant protein fragment. Several parameters were tested in order to optimize the sandwich ELISA assay, including concentration of antibodies, type and concentration of blocking agent, and incubation temperatures. Supernatants from 42 STEC strains of different serotypes and stx variants, including stx2EDL933, stx2vha, stx2vhb, stx2g, stx1EDL933, and stx1d were tested. All Stx variants were detected by the sandwich ELISA, with a detection limit of 115 ng/ml Stx2. Twenty three strains negative for stx genes, including different bacteria species, showed no activity in Vero cell assay and produced negative results in ELISA, except for two strains. Our results show that anti-Stx2B IgY sandwich ELISA could be used in routine diagnosis as a rapid, specific and economic method for detection of Shiga toxin-producing E. coli. PMID:22919675

  1. Oral administration of specific yolk antibodies (IgY) may prevent Pseudomonas aeruginosa infections in patients with cystic fibrosis: a phase I feasibility study.

    PubMed

    Kollberg, Hans; Carlander, David; Olesen, Hanne; Wejåker, Per-Erik; Johannesson, Marie; Larsson, Anders

    2003-06-01

    Respiratory infection is the major cause of morbidity and mortality in cystic fibrosis (CF) patients. Chronic Pseudomonas aeruginosa (PA) infections ultimately occur in virtually all patients. It is impossible to eradicate PA when a patient has been chronically colonized. Immunotherapy with specific egg-yolk antibodies (IgY) may be an alternative to antibiotics for the prevention of PA infections. We wanted to determine if treatment with specific IgY can prolong the period between the first and the second PA colonization? And long-term, can the treatment diminish the number of positive PA cultures and postpone the onset of chronic colonization? CF patients gargled daily with an IgY-antibody preparation, purified from eggs of hens immunized with PA bacteria. They were compared to a group of patients who did not gargle with the preparation. Both groups had their first colonization with PA eradicated by antibiotics. The basic treatment was essentially the same in both groups. In the initial study, the period between the first and second colonization with PA was significantly prolonged for the treated vs. the control group (Kaplan-Meier P = 0.015, Breslow test). In the prolonged study, the treated group had only 2.5 sputum cultures positive for PA per 100 months of observation, and none of these patients became chronically colonized with PA. No adverse events were reported. In the control group, 13.7 cultures per 100 months of observation were positive for PA, and 5 (24%) patients became chronically colonized with PA. This feasibility study shows that antipseudomonal IgY has the potential to effectively prevent PA colonization without any severe adverse effects. A phase III study should be initiated. PMID:12746939

  2. Replacement of a Björk-Shiley Delrin Aortic Valve Still Functioning after 25 Years

    PubMed Central

    Badak, M. Ismail; Ozkisacik, Erdem Ali; Boga, Mehmet; Gurcun, Ugur; Discigil, Berent

    2004-01-01

    We report the case of a patient who had undergone implantation of a Björk-Shiley Delrin valve in the aortic position 25 years earlier and who now presented with severe mitral stenosis. The patient underwent mitral valve replacement and aortic valve re-replacement. We review the justification for prophylactic replacement of Björk-Shiley Delrin heart valves. PMID:15562853

  3. Rk1, a ginsenoside, is a new blocker of vascular leakage acting through actin structure remodeling.

    PubMed

    Maeng, Yong-Sun; Maharjan, Sony; Kim, Jeong-Hun; Park, Jeong-Hill; Suk Yu, Young; Kim, Young-Myoung; Kwon, Young-Guen

    2013-01-01

    Endothelial barrier integrity is essential for vascular homeostasis and increased vascular permeability and has been implicated in many pathological processes, including diabetic retinopathy. Here, we investigated the effect of Rk1, a ginsenoside extracted from sun ginseng, on regulation of endothelial barrier function. In human retinal endothelial cells, Rk1 strongly inhibited permeability induced by VEGF, advanced glycation end-product, thrombin, or histamine. Furthermore, Rk1 significantly reduced the vessel leakiness of retina in a diabetic mouse model. This anti-permeability activity of Rk1 is correlated with enhanced stability and positioning of tight junction proteins at the boundary between cells. Signaling experiments revealed that Rk1 induces phosphorylation of myosin light chain and cortactin, which are critical regulators for the formation of the cortical actin ring structure and endothelial barrier. These findings raise the possibility that ginsenoside Rk1 could be exploited as a novel prototype compound for the prevention of human diseases that are characterized by vascular leakage. PMID:23894330

  4. SnRK1A-interacting negative regulators modulate the nutrient starvation signaling sensor SnRK1 in source-sink communication in cereal seedlings under abiotic stress.

    PubMed

    Lin, Chien-Ru; Lee, Kuo-Wei; Chen, Chih-Yu; Hong, Ya-Fang; Chen, Jyh-Long; Lu, Chung-An; Chen, Ku-Ting; Ho, Tuan-Hua David; Yu, Su-May

    2014-02-01

    In plants, source-sink communication plays a pivotal role in crop productivity, yet the underlying regulatory mechanisms are largely unknown. The SnRK1A protein kinase and transcription factor MYBS1 regulate the sugar starvation signaling pathway during seedling growth in cereals. Here, we identified plant-specific SnRK1A-interacting negative regulators (SKINs). SKINs antagonize the function of SnRK1A, and the highly conserved GKSKSF domain is essential for SKINs to function as repressors. Overexpression of SKINs inhibits the expression of MYBS1 and hydrolases essential for mobilization of nutrient reserves in the endosperm, leading to inhibition of seedling growth. The expression of SKINs is highly inducible by drought and moderately by various stresses, which is likely related to the abscisic acid (ABA)-mediated repression of SnRK1A under stress. Overexpression of SKINs enhances ABA sensitivity for inhibition of seedling growth. ABA promotes the interaction between SnRK1A and SKINs and shifts the localization of SKINs from the nucleus to the cytoplasm, where it binds SnRK1A and prevents SnRK1A and MYBS1 from entering the nucleus. Our findings demonstrate that SnRK1A plays a key role regulating source-sink communication during seedling growth. Under abiotic stress, SKINs antagonize the function of SnRK1A, which is likely a key factor restricting seedling vigor. PMID:24569770

  5. Electrochemical immunosensor based on ensemble of nanoelectrodes for immunoglobulin IgY detection: application to identify hen's egg yolk in tempera paintings.

    PubMed

    Bottari, Fabio; Oliveri, Paolo; Ugo, Paolo

    2014-02-15

    A nanostructured electrochemical biosensor for detecting proteins of interest in work of art, in particular in tempera paintings, is presented. To determine egg yolk we focus here on the determination of immunoglobulin IgY. The transducers are nanoelectrode ensembles (NEEs), prepared via membrane templated electroless deposition of gold. Because of their geometrical and diffusion characteristics, NEEs are characterized by significantly low detection limits, moreover they display the capability of capturing proteins by interaction with the polycarbonate membrane of the NEE. At first, the proteic component of the paint is extracted by ultrasonication in an aqueous buffer, then IgY is captured by incubation on the NEE. The immunoglobulin is detected by treatment with anti-IgY labeled with horse radish peroxidase (Anti-IgY-HRP). The binding of the Anti-IgY-HRP is detected by recording the electrocatalytic signal caused by addition of H2O2 and methylene blue. The sensor detection capabilities are tested by analyzing both paint models, prepared in the lab, and real samples, from paintings of the XVIII-XX century. Multivariate exploratory analysis is applied to classify the voltammetric patterns, confirming the capability to differentiate egg-yolk tempera from other kind of tempera binders as well as from acrylic or oil paints. PMID:24096186

  6. CAD techniques applied to diesel engine design. Extension of the RK range. [Ruston diesels

    SciTech Connect

    Sinha, S.K.; Buckthorpe, D.E.

    1980-01-01

    Rustion Diesels Ltd. produce three ranges of engines, the AP range covering engine powers from 500 to 1400 bhp (350 to 1000 kW electrical), the RK range covering 1410 to 4200 bhp (1 to 3 MW electrical), and the AT range covering 1650 to 4950 bhp (1-2 to 3-5 MW electrical). The AT engine range is available at speeds up to 600 rev/min, whereas the AP and RK ranges cover engine speeds from 600 to 1000 rev/min. The design philosophy and extension of the RK range of engines are investigated. This is a 251 mm (ten inch) bore by 305mm (twelve inch) stroke engine and is available in 6-cylinder in-line form and 8-, 12-, and 16-cylinder vee form. The RK engine features a cast-iron crankcase and bedplate design with a forged alloy-steel crankshaft. Combustion-chamber components consist of a cast-iron cylinder head and liner, steel exhaust and inlet valves, and a single-piece aluminium piston. The durability and reliability of RK engines have been fully proven in service with over 30 years' experience in numerous applications for power generation, reaction, and marine propulsion.

  7. Molecular Insights into the Enigmatic Metabolic Regulator, SnRK1.

    PubMed

    Emanuelle, Shane; Doblin, Monika S; Stapleton, David I; Bacic, Antony; Gooley, Paul R

    2016-04-01

    Sucrose non-fermenting-1 (SNF1)-related kinase 1 (SnRK1) lies at the heart of metabolic homeostasis in plants and is crucial for normal development and response to stress. Evolutionarily related to SNF1 in yeast and AMP-activated kinase (AMPK) in mammals, SnRK1 acts protectively to maintain homeostasis in the face of fluctuations in energy status. Despite a conserved function, the structure and regulation of the plant kinase differ considerably from its relatively well-understood opisthokont orthologues. In this review, we highlight the known plant-specific modes of regulation involving SnRK1 together with new insights based on a 3D molecular model of the kinase. We also summarise how these differences from other orthologues may be specific adaptations to plant metabolism, and offer insights into possible avenues of future inquiry into this enigmatic enzyme. PMID:26642889

  8. Complete Genome Sequence of Noncytopathic Bovine Viral Diarrhea Virus 1 Contaminating a High-Passage RK-13 Cell Line

    PubMed Central

    Nam, Bora; Li, Ganwu; Zheng, Ying; Zhang, Jianqiang; Shuck, Kathleen M.; Timoney, Peter J.

    2015-01-01

    A high-passage rabbit kidney RK-13 cell line (HP-RK-13[KY], originally derived from the ATCC CCL-37 cell line) used in certain laboratories worldwide is contaminated with noncytopathic bovine viral diarrhea virus (ncpBVDV). On complete genome sequence analysis, the virus strain was found to belong to BVDV group 1b. PMID:26430037

  9. Structural basis for basal activity and autoactivation of abscisic acid (ABA) signaling SnRK2 kinases

    SciTech Connect

    Ng, Ley-Moy; Soon, Fen-Fen; Zhou, X. Edward; West, Graham M.; Kovach, Amanda; Suino-Powell, Kelly M.; Chalmers, Michael J.; Li, Jun; Yong, Eu-Leong; Zhu, Jian-Kang; Griffin, Patrick R.; Melcher, Karsten; Xu, H. Eric

    2014-10-02

    Abscisic acid (ABA) is an essential hormone that controls plant growth, development, and responses to abiotic stresses. Central for ABA signaling is the ABA-mediated autoactivation of three monomeric Snf1-related kinases (SnRK2.2, -2.3, and -2.6). In the absence of ABA, SnRK2s are kept in an inactive state by forming physical complexes with type 2C protein phosphatases (PP2Cs). Upon relief of this inhibition, SnRK2 kinases can autoactivate through unknown mechanisms. Here, we report the crystal structures of full-length Arabidopsis thaliana SnRK2.3 and SnRK2.6 at 1.9- and 2.3-{angstrom} resolution, respectively. The structures, in combination with biochemical studies, reveal a two-step mechanism of intramolecular kinase activation that resembles the intermolecular activation of cyclin-dependent kinases. First, release of inhibition by PP2C allows the SnRK2s to become partially active because of an intramolecular stabilization of the catalytic domain by a conserved helix in the kinase regulatory domain. This stabilization enables SnRK2s to gain full activity by activation loop autophosphorylation. Autophosphorylation is more efficient in SnRK2.6, which has higher stability than SnRK2.3 and has well-structured activation loop phosphate acceptor sites that are positioned next to the catalytic site. Together, these data provide a structural framework that links ABA-mediated release of PP2C inhibition to activation of SnRK2 kinases.

  10. High resolution fluorescence microscopy evidence on the transport of immunoglobulins. Differences between mammalian IgG, F(ab')2 and avian IgY.

    PubMed

    Sevcik, Carlos; Salazar, Víctor; Díaz, Patricia; D'Suze, Gina; Vázquez, Hilda

    2013-03-01

    We describe the subcellular localization of horse F(ab')(2) and IgG, and ostrich IgY labeled with fluorescein isothiocyanate (FITC) administered IV to mice. We used wide field high sensitivity fluorescence microscopy deblurred by 3-dimensional blind deconvolution of kidney, liver, lungs and brain sections. Sections were obtained from mice sacrificed 15 min, 1 or 5 h after receiving FITC-immunoproteins, counter-stained with DAPI (4',6'-diamidino-2-phenylindole) and Evans blue. FITC-IgG and its fractions are rapidly taken up and extravasated by vascular endothelium. FITC-IgG and FITC-F(ab')(2) appear to be quickly secreted by glomeruli endothelium and to be reabsorbed along all nephron segments. FITC-IgG and FITC-F(ab')(2) appeared 15 min after IV injection within bronchial, alveolar and bile duct epithelium. Hepatocytes were loaded with fluorescence after 15 min of administration. Fluorescence was absent from brain slices, except for the endothelium of some vessels in brain ventricles which appeared intensely fluorescent. Fluorescence appeared in intracellular vesicles which conferred the tissues a glowing foamy aspect for up to 5 h after inoculation. Arterial elastic layers were intensely green after horse FITC-Ig inoculation. Ostrich FITC-IgY behaved completely differently to horse Ig's; only 1 h after injection it was possible to observe small brightly green scarce vesicles in vascular endothelium of arteries, interstitial kidney capillaries between nephron tubules and were also scarce in glomeruli endothelium; FITC-IgY appeared only in hepatic sinusoids in the liver. No IgY was seen in bronchial and alveolar endothelium, in bile ducts or in hepatocytes. PMID:23159396

  11. SMiRK: an Automated Pipeline for miRNA Analysis

    PubMed Central

    Milholland, Brandon; Gombar, Saurabh; Suh, Yousin

    2015-01-01

    Background Micro RNAs (miRNAs), important regulators of cell function, can be interrogated by high-throughput sequencing in a rapid and cost-effective manner. However, the tremendous amount of data generated by such methods is not easily analyzed. In order to extract meaningful information and draw biological conclusions from miRNA data, many challenges in quality control, alignment, normalization, and analysis must be overcome. Typically, these would only be possible with the dedicated efforts of a specialized computational biologist for a sustained period of time. Results Here, we present SMiRK, an automated pipeline that allows such tasks to be completed with minimal time and without dedicated bioinformatics personnel. SMiRK’s flexibility also allows experienced users to exert more control, if they wish. We describe how SMiRK automatically normalizes the data, removes low-information miRNAs, and produces heatmaps of the processed data. We give details on SMiRK’s implementation and use cases for novice and advanced users. As a demonstration of its capabilities, SMiRK was used to rapidly and automatically analyze a dataset taken from the literature. Conclusion SMiRK is a useful and efficient tool that can be used by investigators at multiple skill levels. Those who lack bioinformatics training can use it to easily and automatically analyze their data, while those with experience will find it beneficial to not need to write tools from scratch. PMID:26613105

  12. Draft Genome Sequence of Ustilago trichophora RK089, a Promising Malic Acid Producer

    PubMed Central

    Zambanini, Thiemo; Buescher, Joerg M.; Meurer, Guido; Blank, Lars M.

    2016-01-01

    The basidiomycetous smut fungus Ustilago trichophora RK089 produces malate from glycerol. De novo genome sequencing revealed a 20.7-Mbp genome (301 gap-closed contigs, 246 scaffolds). A comparison to the genome of Ustilago maydis 521 revealed all essential genes for malate production from glycerol contributing to metabolic engineering for improving malate production. PMID:27469969

  13. Draft Genome Sequence of Ustilago trichophora RK089, a Promising Malic Acid Producer.

    PubMed

    Zambanini, Thiemo; Buescher, Joerg M; Meurer, Guido; Wierckx, Nick; Blank, Lars M

    2016-01-01

    The basidiomycetous smut fungus Ustilago trichophora RK089 produces malate from glycerol. De novo genome sequencing revealed a 20.7-Mbp genome (301 gap-closed contigs, 246 scaffolds). A comparison to the genome of Ustilago maydis 521 revealed all essential genes for malate production from glycerol contributing to metabolic engineering for improving malate production. PMID:27469969

  14. The complex becomes more complex: protein-protein interactions of SnRK1 with DUF581 family proteins provide a framework for cell- and stimulus type-specific SnRK1 signaling in plants

    PubMed Central

    Nietzsche, Madlen; Schießl, Ingrid; Börnke, Frederik

    2014-01-01

    In plants, SNF1-related kinase (SnRK1) responds to the availability of carbohydrates as well as to environmental stresses by down-regulating ATP consuming biosynthetic processes, while stimulating energy-generating catabolic reactions through gene expression and post-transcriptional regulation. The functional SnRK1 complex is a heterotrimer where the catalytic α subunit associates with a regulatory β subunit and an activating γ subunit. Several different metabolites as well as the hormone abscisic acid (ABA) have been shown to modulate SnRK1 activity in a cell- and stimulus-type specific manner. It has been proposed that tissue- or stimulus-specific expression of adapter proteins mediating SnRK1 regulation can at least partly explain the differences observed in SnRK1 signaling. By using yeast two-hybrid and in planta bi-molecular fluorescence complementation assays we were able to demonstrate that proteins containing the domain of unknown function (DUF) 581 could interact with both isoforms of the SnRK1α subunit (AKIN10/11) of Arabidopsis. A structure/function analysis suggests that the DUF581 is a generic SnRK1 interaction module and co-expression with DUF581 proteins in plant cells leads to reallocation of the kinase to specific regions within the nucleus. Yeast two-hybrid analyses suggest that SnRK1 and DUF581 proteins share common interaction partners inside the nucleus. The analysis of available microarray data implies that expression of the 19 members of the DUF581 encoding gene family in Arabidopsis is differentially regulated by hormones and environmental cues, indicating specialized functions of individual family members. We hypothesize that DUF581 proteins could act as mediators conferring tissue- and stimulus-type specific differences in SnRK1 regulation. PMID:24600465

  15. SnRK1 Phosphorylation of AL2 Delays Cabbage Leaf Curl Virus Infection in Arabidopsis

    PubMed Central

    Shen, Wei; Dallas, Mary Beth; Goshe, Michael B.

    2014-01-01

    ABSTRACT Geminivirus AL2/C2 proteins play key roles in establishing infection and causing disease in their plant hosts. They are involved in viral gene expression, counter host defenses by suppressing transcriptional gene silencing, and interfere with the host signaling involved in pathogen resistance. We report here that begomovirus and curtovirus AL2/C2 proteins interact strongly with host geminivirus Rep-interacting kinases (GRIKs), which are upstream activating kinases of the protein kinase SnRK1, a global regulator of energy and nutrient levels in plants. We used an in vitro kinase system to show that GRIK-activated SnRK1 phosphorylates recombinant AL2/C2 proteins from several begomoviruses and to map the SnRK1 phosphorylation site to serine-109 in the AL2 proteins of two New World begomoviruses: Cabbage Leaf Curl Virus (CaLCuV) and Tomato mottle virus. A CaLCuV AL2 S109D phosphomimic mutation did not alter viral DNA levels in protoplast replication assays. In contrast, the phosphomimic mutant was delayed for symptom development and viral DNA accumulation during infection of Arabidopsis thaliana, demonstrating that SnRK1 contributes to host defenses against CaLCuV. Our observation that serine-109 is not conserved in all AL2/C2 proteins that are SnRK1 substrates in vitro suggested that phosphorylation of viral proteins by plant kinases contributes to the evolution of geminivirus-host interactions. IMPORTANCE Geminiviruses are single-stranded DNA viruses that cause serious diseases in many crops. Dicot-infecting geminiviruses carry genes that encode multifunctional AL2/C2 proteins that are essential for infection. However, it is not clear how AL2/C2 proteins are regulated. Here, we show that the host protein kinase SnRK1, a central regulator of energy balance and nutrient metabolism in plants, phosphorylates serine-109 in AL2 proteins of three subgroups of New World begomoviruses, resulting in a delay in viral DNA accumulation and symptom appearance. Our results

  16. miRNAs mediate SnRK1-dependent energy signaling in Arabidopsis

    PubMed Central

    Confraria, Ana; Martinho, Cláudia; Elias, Alexandre; Rubio-Somoza, Ignacio; Baena-González, Elena

    2013-01-01

    The SnRK1 protein kinase, the plant ortholog of mammalian AMPK and yeast Snf1, is activated by the energy depletion caused by adverse environmental conditions. Upon activation, SnRK1 triggers extensive transcriptional changes to restore homeostasis and promote stress tolerance and survival partly through the inhibition of anabolism and the activation of catabolism. Despite the identification of a few bZIP transcription factors as downstream effectors, the mechanisms underlying gene regulation, and in particular gene repression by SnRK1, remain mostly unknown. microRNAs (miRNAs) are 20–24 nt RNAs that regulate gene expression post-transcriptionally by driving the cleavage and/or translation attenuation of complementary mRNA targets. In addition to their role in plant development, mounting evidence implicates miRNAs in the response to environmental stress. Given the involvement of miRNAs in stress responses and the fact that some of the SnRK1-regulated genes are miRNA targets, we postulated that miRNAs drive part of the transcriptional reprogramming triggered by SnRK1. By comparing the transcriptional response to energy deprivation between WT and dcl1-9, a mutant deficient in miRNA biogenesis, we identified 831 starvation genes misregulated in the dcl1-9 mutant, out of which 155 are validated or predicted miRNA targets. Functional clustering analysis revealed that the main cellular processes potentially co-regulated by SnRK1 and miRNAs are translation and organelle function and uncover TCP transcription factors as one of the most highly enriched functional clusters. TCP repression during energy deprivation was impaired in miR319 knockdown (MIM319) plants, demonstrating the involvement of miR319 in the stress-dependent regulation of TCPs. Altogether, our data indicates that miRNAs are components of the SnRK1 signaling cascade contributing to the regulation of specific mRNA targets and possibly tuning down particular cellular processes during the stress response

  17. Detergent-like activity and alpha-helical structure of warnericin RK, an anti-Legionella peptide.

    PubMed

    Verdon, Julien; Falge, Mirjam; Maier, Elke; Bruhn, Heike; Steinert, Michael; Faber, Cornelius; Benz, Roland; Héchard, Yann

    2009-10-01

    Warnericin RK is the first antimicrobial peptide known to be active against Legionella pneumophila, a pathogen bacterium that is responsible for severe pneumonia. Strikingly, this peptide displays a very narrow range of antimicrobial activity, almost limited to the Legionella genus, and a hemolytic activity. A similar activity has been described for delta-lysin, a well-known hemolytic peptide of Staphylococci that has not been described as antimicrobial. In this study we aimed to understand the mode of action of warnericin RK and to explain its particular target specificity. We found that warnericin RK permeabilizes artificial membranes in a voltage-independent manner. Osmotic protection experiments on erythrocytes showed that warnericin RK does not form well-defined pores, suggesting a detergent-like mode of action, as previously described for delta-lysin at high concentrations. Warnericin RK also permeabilized Legionella cells, and these cells displayed a high sensitivity to detergents. Depending on the detergent used, Legionella was from 10- to 1000-fold more sensitive than the other bacteria tested. Finally, the structure of warnericin RK was investigated by means of circular dichroism and NMR spectroscopy. The peptide adopted an amphiphilic alpha-helical structure, consistent with the proposed mode of action. We conclude that the specificity of warnericin RK toward Legionella results from both the detergent-like mode of action of the peptide and the high sensitivity of these bacteria to detergents. PMID:19804724

  18. Detergent-Like Activity and α-Helical Structure of Warnericin RK, an Anti-Legionella Peptide

    PubMed Central

    Verdon, Julien; Falge, Mirjam; Maier, Elke; Bruhn, Heike; Steinert, Michael; Faber, Cornelius; Benz, Roland; Héchard, Yann

    2009-01-01

    Abstract Warnericin RK is the first antimicrobial peptide known to be active against Legionella pneumophila, a pathogen bacterium that is responsible for severe pneumonia. Strikingly, this peptide displays a very narrow range of antimicrobial activity, almost limited to the Legionella genus, and a hemolytic activity. A similar activity has been described for δ-lysin, a well-known hemolytic peptide of Staphylococci that has not been described as antimicrobial. In this study we aimed to understand the mode of action of warnericin RK and to explain its particular target specificity. We found that warnericin RK permeabilizes artificial membranes in a voltage-independent manner. Osmotic protection experiments on erythrocytes showed that warnericin RK does not form well-defined pores, suggesting a detergent-like mode of action, as previously described for δ-lysin at high concentrations. Warnericin RK also permeabilized Legionella cells, and these cells displayed a high sensitivity to detergents. Depending on the detergent used, Legionella was from 10- to 1000-fold more sensitive than the other bacteria tested. Finally, the structure of warnericin RK was investigated by means of circular dichroism and NMR spectroscopy. The peptide adopted an amphiphilic α-helical structure, consistent with the proposed mode of action. We conclude that the specificity of warnericin RK toward Legionella results from both the detergent-like mode of action of the peptide and the high sensitivity of these bacteria to detergents. PMID:19804724

  19. Fatty acid composition modulates sensitivity of Legionella pneumophila to warnericin RK, an antimicrobial peptide.

    PubMed

    Verdon, Julien; Labanowski, Jérome; Sahr, Tobias; Ferreira, Thierry; Lacombe, Christian; Buchrieser, Carmen; Berjeaud, Jean-Marc; Héchard, Yann

    2011-04-01

    Warnericin RK is an antimicrobial peptide, produced by a Staphyloccocus warneri strain, described to be specifically active against Legionella, the pathogenic bacteria responsible for Legionnaires' disease. Warnericin RK is an amphiphilic alpha-helical peptide, which possesses a detergent-like mode of action. Two others peptides, δ-hemolysin I and II, produced by the same S. warneri strain, are highly similar to S. aureus δ-hemolysin and also display anti-Legionella activity. It has been recently reported that S. aureus δ-hemolysin activity on vesicles is likewise related to phospholipid acyl-chain structure, such as chain length and saturation. As staphylococcal δ-hemolysins were highly similar, we thus hypothesized that fatty acid composition of Legionella's membrane might influence the sensitivity of the bacteria to warnericin RK. Relationship between sensitivity to the peptide and fatty acid composition was then followed in various conditions. Cells in stationary phase, which were already described as less resistant than cells in exponential phase, displayed higher amounts of branched-chain fatty acids (BCFA) and short chain fatty acids. An adapted strain, able to grow at a concentration 33 fold higher than minimal inhibitory concentration of the wild type (i.e. 1μM), was isolated after repeated transfers of L. pneumophila in the presence of increased concentrations of warnericin RK. The amount of BCFA was significantly higher in the adapted strain than in the wild type strain. Also, a transcriptomic analysis of the wild type and adapted strains showed that two genes involved in fatty acid biosynthesis were repressed in the adapted strain. These genes encode enzymes involved in desaturation and elongation of fatty acids respectively. Their repression was in agreement with the decrease of unsaturated fatty acids and fatty acid chain length in the adapted strain. Conclusively, our results indicate that the increase of BCFA and the decrease of fatty acid

  20. Characterization of a common wheat (Triticum aestivum L.) TaSnRK2.7 gene involved in abiotic stress responses

    PubMed Central

    Zhang, Hongying; Mao, Xinguo; Jing, Ruilian; Chang, Xiaoping; Xie, Huimin

    2011-01-01

    Sucrose non-fermenting-1-related protein kinase 2 (SnRK2) plays a key role in the plant stress signalling transduction pathway via phosphorylation. Here, a SnRK2 member of common wheat, TaSnRK2.7, was cloned and characterized. Southern blot analysis suggested that the common wheat genome contains three copies of TaSnRK2.7. Subcellular localization showed the presence of TaSnRK2.7 in the cell membrane, cytoplasm, and nucleus. Expression patterns revealed that TaSnRK2.7 is expressed strongly in roots, and responds to polyethylene glycol, NaCl, and cold stress, but not to abscisic acid (ABA) application, suggesting that TaSnRK2.7 might participate in non-ABA-dependent signal transduction pathways. TaSnRK2.7 was transferred to Arabidopsis under the control of the CaMV-35S promoter. Function analysis showed that TaSnRK2.7 is involved in carbohydrate metabolism, decreasing osmotic potential, enhancing photosystem II activity, and promoting root growth. Its overexpression results in enhanced tolerance to multi-abiotic stress. Therefore, TaSnRK2.7 is a multifunctional regulatory factor in plants, and has the potential to be utilized in transgenic breeding to improve abiotic stress tolerance in crop plants. PMID:21030389

  1. Mechanisms of regulation of SNF1/AMPK/SnRK1 protein kinases

    PubMed Central

    Crozet, Pierre; Margalha, Leonor; Confraria, Ana; Rodrigues, Américo; Martinho, Cláudia; Adamo, Mattia; Elias, Carlos A.; Baena-González, Elena

    2014-01-01

    The SNF1 (sucrose non-fermenting 1)-related protein kinases 1 (SnRKs1) are the plant orthologs of the budding yeast SNF1 and mammalian AMPK (AMP-activated protein kinase). These evolutionarily conserved kinases are metabolic sensors that undergo activation in response to declining energy levels. Upon activation, SNF1/AMPK/SnRK1 kinases trigger a vast transcriptional and metabolic reprograming that restores energy homeostasis and promotes tolerance to adverse conditions, partly through an induction of catabolic processes and a general repression of anabolism. These kinases typically function as a heterotrimeric complex composed of two regulatory subunits, β and γ, and an α-catalytic subunit, which requires phosphorylation of a conserved activation loop residue for activity. Additionally, SNF1/AMPK/SnRK1 kinases are controlled by multiple mechanisms that have an impact on kinase activity, stability, and/or subcellular localization. Here we will review current knowledge on the regulation of SNF1/AMPK/SnRK1 by upstream components, post-translational modifications, various metabolites, hormones, and others, in an attempt to highlight both the commonalities of these essential eukaryotic kinases and the divergences that have evolved to cope with the particularities of each one of these systems. PMID:24904600

  2. Mechanisms of regulation of SNF1/AMPK/SnRK1 protein kinases.

    PubMed

    Crozet, Pierre; Margalha, Leonor; Confraria, Ana; Rodrigues, Américo; Martinho, Cláudia; Adamo, Mattia; Elias, Carlos A; Baena-González, Elena

    2014-01-01

    The SNF1 (sucrose non-fermenting 1)-related protein kinases 1 (SnRKs1) are the plant orthologs of the budding yeast SNF1 and mammalian AMPK (AMP-activated protein kinase). These evolutionarily conserved kinases are metabolic sensors that undergo activation in response to declining energy levels. Upon activation, SNF1/AMPK/SnRK1 kinases trigger a vast transcriptional and metabolic reprograming that restores energy homeostasis and promotes tolerance to adverse conditions, partly through an induction of catabolic processes and a general repression of anabolism. These kinases typically function as a heterotrimeric complex composed of two regulatory subunits, β and γ, and an α-catalytic subunit, which requires phosphorylation of a conserved activation loop residue for activity. Additionally, SNF1/AMPK/SnRK1 kinases are controlled by multiple mechanisms that have an impact on kinase activity, stability, and/or subcellular localization. Here we will review current knowledge on the regulation of SNF1/AMPK/SnRK1 by upstream components, post-translational modifications, various metabolites, hormones, and others, in an attempt to highlight both the commonalities of these essential eukaryotic kinases and the divergences that have evolved to cope with the particularities of each one of these systems. PMID:24904600

  3. Accelerated solution of non-linear flow problems using Chebyshev iteration polynomial based RK recursions

    SciTech Connect

    Lorber, A.A.; Carey, G.F.; Bova, S.W.; Harle, C.H.

    1996-12-31

    The connection between the solution of linear systems of equations by iterative methods and explicit time stepping techniques is used to accelerate to steady state the solution of ODE systems arising from discretized PDEs which may involve either physical or artificial transient terms. Specifically, a class of Runge-Kutta (RK) time integration schemes with extended stability domains has been used to develop recursion formulas which lead to accelerated iterative performance. The coefficients for the RK schemes are chosen based on the theory of Chebyshev iteration polynomials in conjunction with a local linear stability analysis. We refer to these schemes as Chebyshev Parameterized Runge Kutta (CPRK) methods. CPRK methods of one to four stages are derived as functions of the parameters which describe an ellipse {Epsilon} which the stability domain of the methods is known to contain. Of particular interest are two-stage, first-order CPRK and four-stage, first-order methods. It is found that the former method can be identified with any two-stage RK method through the correct choice of parameters. The latter method is found to have a wide range of stability domains, with a maximum extension of 32 along the real axis. Recursion performance results are presented below for a model linear convection-diffusion problem as well as non-linear fluid flow problems discretized by both finite-difference and finite-element methods.

  4. The rK39 strip test is non-predictor of clinical status for kala-azar

    PubMed Central

    Singh, Dharmendra P; Sundar, Shyam; Mohapatra, Tribhuban M

    2009-01-01

    Background The rK39 strip test is reported to be simple, sensitive, specific, non-invasive and economical test. Since this method is supposed to be patient friendly, it may easily be accepted for sero-epidemiological surveys. An attempt was made to evaluate the role of rK39 strip test in pre and post treatment phases of Kala azar, as a diagnostic and prognostic marker, in addition to other laboratory investigations, in order to evaluate its role in sero-epidemiological surveys. Findings A total of 210 cases were selected for the study. One hundred clinically and parasitologically confirmed cases were corroborated with other hematological profiles. The formol-gel test was included along with well matched control group comprising of normal endemic controls (50), non-endemic normals (20) and other febrile cases (40). All groups were tested by rK39 strip test. Fifty Kala azar cases were followed up after completion of successful treatment. They were subjected to rK39 strip test after 0, 90 and 180 days of completion of successful treatment. The rK39 showed sensitivity, specificity, PPV, NPV, and diagnostic accuracy of 98% (95% CI 91.7-100), 100%, 100%, 90% (95% CI 66-100) and 98% (95% CI 92.6-100) respectively. All the 50 cured followed up cases showed positive result by rK39 strip test even after 180 days of completion of successful treatment. Conclusion The test seems an ideal qualitative test for the diagnosis of kala-azar. But for sero-epidemiological studies the test may be used with other parameters. Alternatively a quantitative ELISA using rK39 antigen may be used. PMID:19772616

  5. Björk-Jarabak cephalometric analysis on CBCT synthesized cephalograms with different dentofacial sagittal skeletal patterns

    PubMed Central

    Rodriguez-Cardenas, Yalil Augusto; Arriola-Guillen, Luis Ernesto; Flores-Mir, Carlos

    2014-01-01

    OBJECTIVE: The objective of this study was to evaluate the Björk and Jabarak cephalometric analysis generated from cone-beam computed tomography (CBCT) synthesized lateral cephalograms in adults with different sagittal skeletal patterns. METHODS: The sample consisted of 46 CBCT synthesized cephalograms obtained from patients between 16 and 40 years old. A Björk and Jarabak cephalometric analysis among different sagittal skeletal classes was performed. Analysis of variance (ANOVA), multiple range test of Tukey, Kruskal-Wallis test, and independent t-test were used as appropriate. RESULTS: In comparison to the standard values: Skeletal Class III had increased gonial and superior gonial angles (P < 0.001). This trend was also evident when sex was considered. For Class I males, the sella angle was decreased (P = 0.041), articular angle increased (P = 0.027) and gonial angle decreased (P = 0.002); whereas for Class III males, the gonial angle was increased (P = 0.012). For Class I females, the articular angle was increased (P = 0.029) and the gonial angle decreased (P = 0.004). Björk's sum and Björk and Jabarak polygon sum showed no significant differences. The facial biotype presented in the three sagittal classes was mainly hypodivergent and neutral. CONCLUSIONS: In this sample, skeletal Class III malocclusion was strongly differentiated from the other sagittal classes, specifically in the mandible, as calculated through Björk and Jarabak analysis. PMID:25628079

  6. Molecular evolution, characterization, and expression analysis of SnRK2 gene family in Pak-choi (Brassica rapa ssp. chinensis)

    PubMed Central

    Huang, Zhinan; Tang, Jun; Duan, Weike; Wang, Zhen; Song, Xiaoming; Hou, Xilin

    2015-01-01

    The sucrose non-fermenting 1-related protein kinase 2 (SnRK2) family members are plant-specific serine/threonine kinases that are involved in the plant response to abiotic stress and abscisic acid (ABA)-dependent plant development. Further understanding of the evolutionary history and expression characteristics of these genes will help to elucidate the mechanisms of the stress tolerance in Pak-choi, an important green leafy vegetable in China. Thus, we investigated the evolutionary patterns, footprints and conservation of SnRK2 genes in selected plants and later cloned and analyzed SnRK2 genes in Pak-choi. We found that this gene family was preferentially retained in Brassicas after the Brassica-Arabidopsis thaliana split. Next, we cloned and sequenced 13 SnRK2 from both cDNA and DNA libraries of stress-induced Pak-choi, which were under conditions of ABA, salinity, cold, heat, and osmotic treatments. Most of the BcSnRK2s have eight exons and could be divided into three groups. The subcellular localization predictions suggested that the putative BcSnRK2 proteins were enriched in the nucleus. The results of an analysis of the expression patterns of the BcSnRK2 genes showed that BcSnRK2 group III genes were robustly induced by ABA treatments. Most of the BcSnRK2 genes were activated by low temperature, and the BcSnRK2.6 genes responded to both ABA and low temperature. In fact, most of the BcSnRK2 genes showed positive or negative regulation under ABA and low temperature treatments, suggesting that they may be global regulators that function at the intersection of multiple signaling pathways to play important roles in Pak-choi stress responses. PMID:26557127

  7. Molecular evolution, characterization, and expression analysis of SnRK2 gene family in Pak-choi (Brassica rapa ssp. chinensis).

    PubMed

    Huang, Zhinan; Tang, Jun; Duan, Weike; Wang, Zhen; Song, Xiaoming; Hou, Xilin

    2015-01-01

    The sucrose non-fermenting 1-related protein kinase 2 (SnRK2) family members are plant-specific serine/threonine kinases that are involved in the plant response to abiotic stress and abscisic acid (ABA)-dependent plant development. Further understanding of the evolutionary history and expression characteristics of these genes will help to elucidate the mechanisms of the stress tolerance in Pak-choi, an important green leafy vegetable in China. Thus, we investigated the evolutionary patterns, footprints and conservation of SnRK2 genes in selected plants and later cloned and analyzed SnRK2 genes in Pak-choi. We found that this gene family was preferentially retained in Brassicas after the Brassica-Arabidopsis thaliana split. Next, we cloned and sequenced 13 SnRK2 from both cDNA and DNA libraries of stress-induced Pak-choi, which were under conditions of ABA, salinity, cold, heat, and osmotic treatments. Most of the BcSnRK2s have eight exons and could be divided into three groups. The subcellular localization predictions suggested that the putative BcSnRK2 proteins were enriched in the nucleus. The results of an analysis of the expression patterns of the BcSnRK2 genes showed that BcSnRK2 group III genes were robustly induced by ABA treatments. Most of the BcSnRK2 genes were activated by low temperature, and the BcSnRK2.6 genes responded to both ABA and low temperature. In fact, most of the BcSnRK2 genes showed positive or negative regulation under ABA and low temperature treatments, suggesting that they may be global regulators that function at the intersection of multiple signaling pathways to play important roles in Pak-choi stress responses. PMID:26557127

  8. Genome-wide identification and homeolog-specific expression analysis of the SnRK2 genes in Brassica napus guard cells.

    PubMed

    Yoo, Mi-Jeong; Ma, Tianyi; Zhu, Ning; Liu, Lihong; Harmon, Alice C; Wang, Qiaomei; Chen, Sixue

    2016-05-01

    Sucrose non-fermenting-1-related protein kinase 2 (SnRK2) proteins constitute a small plant-specific serine/threonine kinase family involved in abscisic acid (ABA) signaling and plant responses to biotic and abiotic stresses. Although SnRK2s have been well-studied in Arabidopsis thaliana, little is known about SnRK2s in Brassica napus. Here we identified 30 putative sequences encoding 10 SnRK2 proteins in the B. napus genome and the expression profiles of a subset of 14 SnRK2 genes in guard cells of B. napus. In agreement with its polyploid origin, B. napus maintains both homeologs from its diploid parents. The results of quantitative real-time PCR (qRT-PCR) and reanalysis of RNA-Seq data showed that certain BnSnRK2 genes were commonly expressed in leaf tissues in different varieties of B. napus. In particular, qRT-PCR results showed that 12 of the 14 BnSnRK2s responded to drought stress in leaves and in ABA-treated guard cells. Among them, BnSnRK2.4 and BnSnRK2.6 were of interest because of their robust responsiveness to ABA treatment and drought stress. Notably, BnSnRK2 genes exhibited up-regulation of different homeologs, particularly in response to abiotic stress. The homeolog expression bias in BnSnRK2 genes suggests that parental origin of genes might be responsible for efficient regulation of stress responses in polyploids. This work has laid a foundation for future functional characterization of the different BnSnKR2 homeologs in B. napus and its parents, especially their functions in guard cell signaling and stress responses. PMID:26898295

  9. Placing Intelligence into an Evolutionary Framework or How "g" Fits into the "r-K" Matrix of Life-History Traits Including Longevity

    ERIC Educational Resources Information Center

    Rushton, J. Philippe

    2004-01-01

    First, I describe why intelligence (Spearman's "g") can only be fully understood through "r-K" theory, which places it into an evolutionary framework along with brain size, longevity, maturation speed, and several other life-history traits. The "r-K" formulation explains why IQ predicts longevity and also why the gap in mortality rates between…

  10. Ectopic Expression of Vaccinia Virus E3 and K3 Cannot Rescue Ectromelia Virus Replication in Rabbit RK13 Cells

    PubMed Central

    Peng, Chen; Rothenburg, Stefan; Hersperger, Adam R.

    2015-01-01

    As a group, poxviruses have been shown to infect a wide variety of animal species. However, there is individual variability in the range of species able to be productively infected. In this study, we observed that ectromelia virus (ECTV) does not replicate efficiently in cultured rabbit RK13 cells. Conversely, vaccinia virus (VACV) replicates well in these cells. Upon infection of RK13 cells, the replication cycle of ECTV is abortive in nature, resulting in a greatly reduced ability to spread among cells in culture. We observed ample levels of early gene expression but reduced detection of virus factories and severely blunted production of enveloped virus at the cell surface. This work focused on two important host range genes, named E3L and K3L, in VACV. Both VACV and ECTV express a functional protein product from the E3L gene, but only VACV contains an intact K3L gene. To better understand the discrepancy in replication capacity of these viruses, we examined the ability of ECTV to replicate in wild-type RK13 cells compared to cells that constitutively express E3 and K3 from VACV. The role these proteins play in the ability of VACV to replicate in RK13 cells was also analyzed to determine their individual contribution to viral replication and PKR activation. Since E3L and K3L are two relevant host range genes, we hypothesized that expression of one or both of them may have a positive impact on the ability of ECTV to replicate in RK13 cells. Using various methods to assess virus growth, we did not detect any significant differences with respect to the replication of ECTV between wild-type RK13 compared to versions of this cell line that stably expressed VACV E3 alone or in combination with K3. Therefore, there remain unanswered questions related to the factors that limit the host range of ECTV. PMID:25734776

  11. Ectopic expression of vaccinia virus E3 and K3 cannot rescue ectromelia virus replication in rabbit RK13 cells.

    PubMed

    Hand, Erin S; Haller, Sherry L; Peng, Chen; Rothenburg, Stefan; Hersperger, Adam R

    2015-01-01

    As a group, poxviruses have been shown to infect a wide variety of animal species. However, there is individual variability in the range of species able to be productively infected. In this study, we observed that ectromelia virus (ECTV) does not replicate efficiently in cultured rabbit RK13 cells. Conversely, vaccinia virus (VACV) replicates well in these cells. Upon infection of RK13 cells, the replication cycle of ECTV is abortive in nature, resulting in a greatly reduced ability to spread among cells in culture. We observed ample levels of early gene expression but reduced detection of virus factories and severely blunted production of enveloped virus at the cell surface. This work focused on two important host range genes, named E3L and K3L, in VACV. Both VACV and ECTV express a functional protein product from the E3L gene, but only VACV contains an intact K3L gene. To better understand the discrepancy in replication capacity of these viruses, we examined the ability of ECTV to replicate in wild-type RK13 cells compared to cells that constitutively express E3 and K3 from VACV. The role these proteins play in the ability of VACV to replicate in RK13 cells was also analyzed to determine their individual contribution to viral replication and PKR activation. Since E3L and K3L are two relevant host range genes, we hypothesized that expression of one or both of them may have a positive impact on the ability of ECTV to replicate in RK13 cells. Using various methods to assess virus growth, we did not detect any significant differences with respect to the replication of ECTV between wild-type RK13 compared to versions of this cell line that stably expressed VACV E3 alone or in combination with K3. Therefore, there remain unanswered questions related to the factors that limit the host range of ECTV. PMID:25734776

  12. TaSnRK2.4, an SNF1-type serine/threonine protein kinase of wheat (Triticum aestivum L.), confers enhanced multistress tolerance in Arabidopsis.

    PubMed

    Mao, Xinguo; Zhang, Hongying; Tian, Shanjun; Chang, Xiaoping; Jing, Ruilian

    2010-03-01

    Osmotic stresses such as drought, salinity, and cold are major environmental factors that limit agricultural productivity worldwide. Protein phosphorylation/dephosphorylation are major signalling events induced by osmotic stress in higher plants. Sucrose non-fermenting 1-related protein kinase2 family members play essential roles in response to hyperosmotic stresses in Arabidopsis, rice, and maize. In this study, the function of TaSnRK2.4 in drought, salt, and freezing stresses in Arabidopsis was characterized. A translational fusion protein of TaSnRK2.4 with green fluorescent protein showed subcellular localization in the cell membrane, cytoplasm, and nucleus. To examine the role of TaSnRK2.4 under various environmental stresses, transgenic Arabidopsis plants overexpressing wheat TaSnRK2.4 under control of the cauliflower mosaic virus 35S promoter were generated. Overexpression of TaSnRK2.4 resulted in delayed seedling establishment, longer primary roots, and higher yield under normal growing conditions. Transgenic Arabidopsis overexpressing TaSnRK2.4 had enhanced tolerance to drought, salt, and freezing stresses, which were simultaneously supported by physiological results, including decreased rate of water loss, enhanced higher relative water content, strengthened cell membrane stability, improved photosynthesis potential, and significantly increased osmotic potential. The results show that TaSnRK2.4 is involved in the regulation of enhanced osmotic potential, growth, and development under both normal and stress conditions, and imply that TaSnRK2.4 is a multifunctional regulatory factor in Arabidopsis. Since the overexpression of TaSnRK2.4 can significantly strengthen tolerance to drought, salt, and freezing stresses and does not retard the growth of transgenic Arabidopsis plants under well-watered conditions, TaSnRK2.4 could be utilized in transgenic breeding to improve abiotic stresses in crops. PMID:20022921

  13. TaSnRK2.4, an SNF1-type serine/threonine protein kinase of wheat (Triticum aestivum L.), confers enhanced multistress tolerance in Arabidopsis

    PubMed Central

    Mao, Xinguo; Zhang, Hongying; Tian, Shanjun; Chang, Xiaoping; Jing, Ruilian

    2010-01-01

    Osmotic stresses such as drought, salinity, and cold are major environmental factors that limit agricultural productivity worldwide. Protein phosphorylation/dephosphorylation are major signalling events induced by osmotic stress in higher plants. Sucrose non-fermenting 1-related protein kinase2 family members play essential roles in response to hyperosmotic stresses in Arabidopsis, rice, and maize. In this study, the function of TaSnRK2.4 in drought, salt, and freezing stresses in Arabidopsis was characterized. A translational fusion protein of TaSnRK2.4 with green fluorescent protein showed subcellular localization in the cell membrane, cytoplasm, and nucleus. To examine the role of TaSnRK2.4 under various environmental stresses, transgenic Arabidopsis plants overexpressing wheat TaSnRK2.4 under control of the cauliflower mosaic virus 35S promoter were generated. Overexpression of TaSnRK2.4 resulted in delayed seedling establishment, longer primary roots, and higher yield under normal growing conditions. Transgenic Arabidopsis overexpressing TaSnRK2.4 had enhanced tolerance to drought, salt, and freezing stresses, which were simultaneously supported by physiological results, including decreased rate of water loss, enhanced higher relative water content, strengthened cell membrane stability, improved photosynthesis potential, and significantly increased osmotic potential. The results show that TaSnRK2.4 is involved in the regulation of enhanced osmotic potential, growth, and development under both normal and stress conditions, and imply that TaSnRK2.4 is a multifunctional regulatory factor in Arabidopsis. Since the overexpression of TaSnRK2.4 can significantly strengthen tolerance to drought, salt, and freezing stresses and does not retard the growth of transgenic Arabidopsis plants under well-watered conditions, TaSnRK2.4 could be utilized in transgenic breeding to improve abiotic stresses in crops. PMID:20022921

  14. The SnRK1 Energy Sensor in Plant Biotic Interactions.

    PubMed

    Hulsmans, Sander; Rodriguez, Marianela; De Coninck, Barbara; Rolland, Filip

    2016-08-01

    Our understanding of plant biotic interactions has grown significantly in recent years with the identification of the mechanisms involved in innate immunity, hormone signaling, and secondary metabolism. The impact of such interactions on primary metabolism and the role of metabolic signals in the response of the plants, however, remain far less explored. The SnRK1 (SNF1-related kinase 1) kinases act as metabolic sensors, integrating very diverse stress conditions, and are key in maintaining energy homeostasis for growth and survival. Consistently, an important role is emerging for these kinases as regulators of biotic stress responses triggered by viral, bacterial, fungal, and oomycete infections as well as by herbivory. While this identifies SnRK1 as a promising target for directed modification or selection for more quantitative and sustainable resistance, its central function also increases the chances of unwanted side effects on growth and fitness, stressing the need for identification and in-depth characterization of the mechanisms and target processes involved. VIDEO ABSTRACT. PMID:27156455

  15. Functional characterization of WalRK: A two-component signal transduction system from Bacillus anthracis.

    PubMed

    Dhiman, Alisha; Bhatnagar, Sonika; Kulshreshtha, Parul; Bhatnagar, Rakesh

    2014-01-01

    Two-component signal transduction systems (TCS), consisting of a sensor histidine protein kinase and its cognate response regulator, are an important mode of environmental sensing in bacteria. Additionally, they have been found to regulate virulence determinants in several pathogens. Bacillus anthracis, the causative agent of anthrax and a bioterrorism agent, harbours 41 pairs of TCS. However, their role in its pathogenicity has remained largely unexplored. Here, we show that WalRK of B. anthracis forms a functional TCS which exhibits some species-specific functions. Biochemical studies showed that domain variants of WalK, the histidine kinase, exhibit classical properties of autophosphorylation and phosphotransfer to its cognate response regulator WalR. Interestingly, these domain variants also show phosphatase activity towards phosphorylated WalR, thereby making WalK a bifunctional histidine kinase/phosphatase. An in silico regulon determination approach, using a consensus binding sequence from Bacillus subtilis, provided a list of 30 genes that could form a putative WalR regulon in B. anthracis. Further, electrophoretic mobility shift assay was used to show direct binding of purified WalR to the upstream regions of three putative regulon candidates, an S-layer protein EA1, a cell division ABC transporter FtsE and a sporulation histidine kinase KinB3. Our work lends insight into the species-specific functions and mode of action of B. anthracis WalRK. PMID:24490131

  16. Reconsideration of r/K Selection Theory Using Stochastic Control Theory and Nonlinear Structured Population Models

    PubMed Central

    Oizumi, Ryo; Kuniya, Toshikazu; Enatsu, Yoichi

    2016-01-01

    Despite the fact that density effects and individual differences in life history are considered to be important for evolution, these factors lead to several difficulties in understanding the evolution of life history, especially when population sizes reach the carrying capacity. r/K selection theory explains what types of life strategies evolve in the presence of density effects and individual differences. However, the relationship between the life schedules of individuals and population size is still unclear, even if the theory can classify life strategies appropriately. To address this issue, we propose a few equations on adaptive life strategies in r/K selection where density effects are absent or present. The equations detail not only the adaptive life history but also the population dynamics. Furthermore, the equations can incorporate temporal individual differences, which are referred to as internal stochasticity. Our framework reveals that maximizing density effects is an evolutionarily stable strategy related to the carrying capacity. A significant consequence of our analysis is that adaptive strategies in both selections maximize an identical function, providing both population growth rate and carrying capacity. We apply our method to an optimal foraging problem in a semelparous species model and demonstrate that the adaptive strategy yields a lower intrinsic growth rate as well as a lower basic reproductive number than those obtained with other strategies. This study proposes that the diversity of life strategies arises due to the effects of density and internal stochasticity. PMID:27336169

  17. Reconsideration of r/K Selection Theory Using Stochastic Control Theory and Nonlinear Structured Population Models.

    PubMed

    Oizumi, Ryo; Kuniya, Toshikazu; Enatsu, Yoichi

    2016-01-01

    Despite the fact that density effects and individual differences in life history are considered to be important for evolution, these factors lead to several difficulties in understanding the evolution of life history, especially when population sizes reach the carrying capacity. r/K selection theory explains what types of life strategies evolve in the presence of density effects and individual differences. However, the relationship between the life schedules of individuals and population size is still unclear, even if the theory can classify life strategies appropriately. To address this issue, we propose a few equations on adaptive life strategies in r/K selection where density effects are absent or present. The equations detail not only the adaptive life history but also the population dynamics. Furthermore, the equations can incorporate temporal individual differences, which are referred to as internal stochasticity. Our framework reveals that maximizing density effects is an evolutionarily stable strategy related to the carrying capacity. A significant consequence of our analysis is that adaptive strategies in both selections maximize an identical function, providing both population growth rate and carrying capacity. We apply our method to an optimal foraging problem in a semelparous species model and demonstrate that the adaptive strategy yields a lower intrinsic growth rate as well as a lower basic reproductive number than those obtained with other strategies. This study proposes that the diversity of life strategies arises due to the effects of density and internal stochasticity. PMID:27336169

  18. The LisRK Signal Transduction System Determines the Sensitivity of Listeria monocytogenes to Nisin and Cephalosporins

    PubMed Central

    Cotter, Paul D.; Guinane, Caitriona M.; Hill, Colin

    2002-01-01

    The Listeria monocytogenes two-component signal transduction system, LisRK, initially identified in strain LO28, plays a significant role in the virulence potential of this important food-borne pathogen. Here, it is shown that, in addition to its major contribution in responding to ethanol, pH, and hydrogen peroxide stresses, LisRK is involved in the ability of the cell to tolerate important antimicrobials used in food and in medicine, e.g., the lantibiotic nisin and the cephalosporin family of antibiotics. A ΔlisK mutant (lacking the LisK histidine kinase sensor component) displays significantly enhanced resistance to the lantibiotic nisin, a greatly enhanced sensitivity to the cephalosporins, and a large reduction in the expression of three genes thought to encode a penicillin-binding protein, another histidine kinase (other than LisK), and a protein of unknown function. Confirmation of the role of LisRK was obtained when the response regulator, LisR, was overexpressed using both constitutive and inducible (nisin-controlled expression) systems. Under these conditions we observed a reversion of the ΔlisK mutant to wild-type growth kinetics in the presence of nisin. It was also found that overexpression of LisR complemented the reduced expression of two of the aforementioned genes. These results demonstrate the important role of LisRK in the response of L. monocytogenes to a number of antimicrobial agents. PMID:12183229

  19. SnRK1-triggered switch of bZIP63 dimerization mediates the low-energy response in plants

    PubMed Central

    Mair, Andrea; Pedrotti, Lorenzo; Wurzinger, Bernhard; Anrather, Dorothea; Simeunovic, Andrea; Weiste, Christoph; Valerio, Concetta; Dietrich, Katrin; Kirchler, Tobias; Nägele, Thomas; Vicente Carbajosa, Jesús; Hanson, Johannes; Baena-González, Elena; Chaban, Christina; Weckwerth, Wolfram; Dröge-Laser, Wolfgang; Teige, Markus

    2015-01-01

    Metabolic adjustment to changing environmental conditions, particularly balancing of growth and defense responses, is crucial for all organisms to survive. The evolutionary conserved AMPK/Snf1/SnRK1 kinases are well-known metabolic master regulators in the low-energy response in animals, yeast and plants. They act at two different levels: by modulating the activity of key metabolic enzymes, and by massive transcriptional reprogramming. While the first part is well established, the latter function is only partially understood in animals and not at all in plants. Here we identified the Arabidopsis transcription factor bZIP63 as key regulator of the starvation response and direct target of the SnRK1 kinase. Phosphorylation of bZIP63 by SnRK1 changed its dimerization preference, thereby affecting target gene expression and ultimately primary metabolism. A bzip63 knock-out mutant exhibited starvation-related phenotypes, which could be functionally complemented by wild type bZIP63, but not by a version harboring point mutations in the identified SnRK1 target sites. DOI: http://dx.doi.org/10.7554/eLife.05828.001 PMID:26263501

  20. Regulation of Bacteriocin Production and Cell Death by the VicRK Signaling System in Streptococcus mutans

    PubMed Central

    Senadheera, D. B.; Cordova, M.; Ayala, E. A.; Chávez de Paz, L. E.; Singh, K.; Downey, J. S.; Svensäter, G.; Goodman, S. D.

    2012-01-01

    The VicRK two-component signaling system modulates biofilm formation, genetic competence, and stress tolerance in Streptococcus mutans. We show here that the VicRK modulates bacteriocin production and cell viability, in part by direct modulation of competence-stimulating peptide (CSP) production in S. mutans. Global transcriptome and real-time transcriptional analysis of the VicK-deficient mutant (SmuvicK) revealed significant modulation of several bacteriocin-related loci, including nlmAB, nlmC, and nlmD (P < 0.001), suggesting a role for the VicRK in producing mutacins IV, V, and VI. Bacteriocin overlay assays revealed an altered ability of the vic mutants to kill related species. Since a well-conserved VicR binding site (TGTWAH-N5-TGTWAH) was identified within the comC coding region, we confirmed VicR binding to this sequence using DNA footprinting. Overexpression of the vic operon caused growth-phase-dependent repression of comC, comDE, and comX. In the vic mutants, transcription of nlmC/cipB encoding mutacin V, previously linked to CSP-dependent cell lysis, as well as expression of its putative immunity factor encoded by immB, were significantly affected relative to the wild type (P < 0.05). In contrast to previous reports that proposed a hyper-resistant phenotype for the VicK mutant in cell viability, the release of extracellular genomic DNA was significantly enhanced in SmuvicK (P < 0.05), likely as a result of increased autolysis compared with the parent. The drastic influence of VicRK on cell viability was also demonstrated using vic mutant biofilms. Taken together, we have identified a novel regulatory link between the VicRK and ComDE systems to modulate bacteriocin production and cell viability of S. mutans. PMID:22228735

  1. Sucrose non-fermenting kinase 1 (SnRK1) coordinates metabolic and hormonal signals during pea cotyledon growth and differentiation.

    PubMed

    Radchuk, Ruslana; Emery, R J Neil; Weier, Diana; Vigeolas, Helene; Geigenberger, Peter; Lunn, John E; Feil, Regina; Weschke, Winfriede; Weber, Hans

    2010-01-01

    Seed development passes through developmental phases such as cell division, differentiation and maturation: each have specific metabolic demands. The ubiquitous sucrose non-fermenting-like kinase (SnRK1) coordinates and adjusts physiological and metabolic demands with growth. In protoplast assays sucrose deprivation and hormone supplementation, such as with auxin and abscisic acid (ABA), stimulate SnRK1-promoter activity. This indicates regulation by nutrients: hormonal crosstalk under conditions of nutrient demand and cell proliferation. SnRK1-repressed pea (Pisum sativum) embryos show lower cytokinin levels and deregulation of cotyledonary establishment and growth, together with downregulated gene expression related to cell proliferation, meristem maintenance and differentiation, leaf formation, and polarity. This suggests that at early stages of seed development SnRK1 regulates coordinated cotyledon emergence and growth via cytokinin-mediated auxin transport and/or distribution. Decreased ABA levels and reduced gene expression, involved in ABA-mediated seed maturation and response to sugars, indicate that SnRK1 is required for ABA synthesis and/or signal transduction at an early stage. Metabolic profiling of SnRK1-repressed embryos revealed lower levels of most organic and amino acids. In contrast, levels of sugars and glycolytic intermediates were higher or unchanged, indicating decreased carbon partitioning into subsequent pathways such as the tricarbonic acid cycle and amino acid biosynthesis. It is hypothesized that SnRK1 mediates the responses to sugar signals required for early cotyledon establishment and patterning. As a result, later maturation and storage activity are strongly impaired. Changes observed in SnRK1-repressed pea seeds provide a framework for how SnRK1 communicates nutrient and hormonal signals from auxins, cytokinins and ABA to control metabolism and development. PMID:19845880

  2. Quantitative phosphoproteomics reveals the role of the AMPK plant ortholog SnRK1 as a metabolic master regulator under energy deprivation.

    PubMed

    Nukarinen, Ella; Nägele, Thomas; Pedrotti, Lorenzo; Wurzinger, Bernhard; Mair, Andrea; Landgraf, Ramona; Börnke, Frederik; Hanson, Johannes; Teige, Markus; Baena-Gonzalez, Elena; Dröge-Laser, Wolfgang; Weckwerth, Wolfram

    2016-01-01

    Since years, research on SnRK1, the major cellular energy sensor in plants, has tried to define its role in energy signalling. However, these attempts were notoriously hampered by the lethality of a complete knockout of SnRK1. Therefore, we generated an inducible amiRNA::SnRK1α2 in a snrk1α1 knock out background (snrk1α1/α2) to abolish SnRK1 activity to understand major systemic functions of SnRK1 signalling under energy deprivation triggered by extended night treatment. We analysed the in vivo phosphoproteome, proteome and metabolome and found that activation of SnRK1 is essential for repression of high energy demanding cell processes such as protein synthesis. The most abundant effect was the constitutively high phosphorylation of ribosomal protein S6 (RPS6) in the snrk1α1/α2 mutant. RPS6 is a major target of TOR signalling and its phosphorylation correlates with translation. Further evidence for an antagonistic SnRK1 and TOR crosstalk comparable to the animal system was demonstrated by the in vivo interaction of SnRK1α1 and RAPTOR1B in the cytosol and by phosphorylation of RAPTOR1B by SnRK1α1 in kinase assays. Moreover, changed levels of phosphorylation states of several chloroplastic proteins in the snrk1α1/α2 mutant indicated an unexpected link to regulation of photosynthesis, the main energy source in plants. PMID:27545962

  3. Quantitative phosphoproteomics reveals the role of the AMPK plant ortholog SnRK1 as a metabolic master regulator under energy deprivation

    PubMed Central

    Nukarinen, Ella; Nägele, Thomas; Pedrotti, Lorenzo; Wurzinger, Bernhard; Mair, Andrea; Landgraf, Ramona; Börnke, Frederik; Hanson, Johannes; Teige, Markus; Baena-Gonzalez, Elena; Dröge-Laser, Wolfgang; Weckwerth, Wolfram

    2016-01-01

    Since years, research on SnRK1, the major cellular energy sensor in plants, has tried to define its role in energy signalling. However, these attempts were notoriously hampered by the lethality of a complete knockout of SnRK1. Therefore, we generated an inducible amiRNA::SnRK1α2 in a snrk1α1 knock out background (snrk1α1/α2) to abolish SnRK1 activity to understand major systemic functions of SnRK1 signalling under energy deprivation triggered by extended night treatment. We analysed the in vivo phosphoproteome, proteome and metabolome and found that activation of SnRK1 is essential for repression of high energy demanding cell processes such as protein synthesis. The most abundant effect was the constitutively high phosphorylation of ribosomal protein S6 (RPS6) in the snrk1α1/α2 mutant. RPS6 is a major target of TOR signalling and its phosphorylation correlates with translation. Further evidence for an antagonistic SnRK1 and TOR crosstalk comparable to the animal system was demonstrated by the in vivo interaction of SnRK1α1 and RAPTOR1B in the cytosol and by phosphorylation of RAPTOR1B by SnRK1α1 in kinase assays. Moreover, changed levels of phosphorylation states of several chloroplastic proteins in the snrk1α1/α2 mutant indicated an unexpected link to regulation of photosynthesis, the main energy source in plants. PMID:27545962

  4. Longitudinal impedance measurement of an RK-TBA induction accelerating gap

    SciTech Connect

    Eylon, S.; Henestroza, E.; Kim, J.-S.; Houck, T.L.; Westenskow, G.A.; Yu, S.S.

    1997-05-01

    Induction accelerating gap designs are being studied for Relativistic Klystron Two-Beam Accelerator (RK-TBA) applications. The accelerating gap has to satisfy the following major requirements: hold-off of the applied accelerating voltage pulse, low transverse impedance to limit beam breakup, low longitudinal impedance at the beam-modulation frequency to minimize power loss. Various gap geometries, materials and novel insulating techniques were explored to optimize the gap design. We report on the experimental effort to evaluate the rf properties of the accelerating gaps in a simple pillbox cavity structure. The experimental cavity setup was designed using the AMOS, MAFIA and URMEL numerical codes. Longitudinal impedance measurements above beam-tube cut-off frequency using a single-wire measuring system are presented.

  5. The legacy of the IGY

    NASA Astrophysics Data System (ADS)

    Friedman, Herbert

    We are now at the point of celebrating three milestones of international cooperation in sun—earth research: the 100th anniversary of the First International Polar Year (1882-1883) the 50th anniversary of the Second Polar Year (1932-1933) and the 25th anniversary of the International Geophysical Year (1957-1958). Credit for the concept of the First Polar Year goes to an Austrian Lieutenant, Karl Weyprecht. He expressed the philosophy of scientific cooperation in the following bold language delivered in a statement to the Hall of the Austrian Academy of Sciences on January 18, 1875:Purely geographical research and Arctic topography, which until now have stood in the foreground of all polar expeditions, must, with respect to the great scientific questions, recede into the background. The answers, though, will occur only when those nations pretending to aspire to the heights of contemporary, cultural endeavor decide, without regard to national rivalry, upon common measures. In order to secure decisive scientific results, we require a series of simultaneous expeditions whose aims must be, through dispersal over several points of the Arctic region and using identical instruments in line with identical instructions, to conduct a simultaneous, year-long series of observations. Only thereby shall we acquire the material for solutions to those great problems of nature that reside in the Arctic ice, and only then shall we earn the reward for those considerable resources that have hitherto been squandered in labor, endeavor, deprivation, and money in the polar region.

  6. Comparison of an rK39 dipstick rapid test with direct agglutination test and splenic aspiration for the diagnosis of kala-azar in Sudan.

    PubMed

    Veeken, Hans; Ritmeijer, Koert; Seaman, Jill; Davidson, Robert

    2003-02-01

    We compared an rK39 dipstick rapid test (Amrad ICT, Australia) with a direct agglutination test (DAT) and splenic aspirate for the diagnosis of kala-azar in 77 patients. The study was carried out under field conditions in an endemic area of north-east Sudan. The sensitivity of the rK39 test compared with splenic aspiration was 92% (46/50), the specificity 59% (16/27), and the positive predictive value 81% (46/57). Compared with the diagnostic protocol used by Médecins sans Frontières, the sensitivity of the rK39 test was 93% (50/54), the specificity 70% (16/23), and the positive predictive value 88% (50/57). Compared with splenic aspirates, the sensitivity of a DAT with a titre > or =1:400 was 100% (50/50), but its specificity only 55% (15/27) and the positive predictive value was 80% (50/62). Using a DAT titre > or =1:6400, the sensitivity was 84% (42/50), the specificity 85% (23/27) and the positive predictive value 91% (42/46). All four patients with DAT titre > or =1:6400 but negative splenic aspirate were also rK39 positive; we consider these are probably 'true' cases of kala-azar, i.e. false negative aspirates, rather than false DAT and rK39 seropositives. There were no false negative DATs (DAT titre < or =1:400 and aspirate positive), but there were four false negative rK39 tests (rK39 negative and aspirate positive). The rK39 dipstick is a good screening test for kala-azar; but further development is required before it can replace the DAT as a diagnostic test in endemic areas of the Sudan. PMID:12581443

  7. Molecular characterization of the mouse In(10)17Rk inversion and identification of a novel muscle-specific gene at the proximal breakpoint.

    PubMed Central

    Benson, Kathleen F; Chada, Kiran

    2002-01-01

    Chromosomal rearrangements provide an important resource for molecular characterization of mutations in the mouse. In(10)17Rk mice contain a paracentric inversion of approximately 50 Mb on chromosome 10. Homozygous In(10)17Rk mice exhibit a pygmy phenotype, suggesting that the distal inversion breakpoint is within the pygmy locus. The pygmy mutation, originally isolated in 1944, is an autosomal recessive trait causing a dwarf phenotype in homozygous mice and has been mapped to the distal region of chromosome 10. The pygmy phenotype has subsequently been shown to result from disruption of the Hmgi-c gene. To identify the In(10)17Rk distal inversion breakpoint, In(10)17Rk DNA was subjected to RFLP analysis with single copy sequences derived from the wild-type pygmy locus. This analysis localized the In(10)17Rk distal inversion breakpoint to intron 3 of Hmgi-c and further study determined that a fusion transcript between novel 5' sequence and exons 4 and 5 of Hmgi-c is created. We employed 5' RACE to isolate the 5' end of the fusion transcript and this sequence was localized to the proximal end of chromosome 10 between markers Cni-rs2 and Mtap7. Northern blot analysis of individual tissues of wild-type mice determined that the gene at the In(10)17Rk proximal inversion breakpoint is a novel muscle-specific gene and its disruption does not lead to a readily observable phenotype. PMID:11805063

  8. A meta-analysis of the diagnostic performance of the direct agglutination test and rK39 dipstick for visceral leishmaniasis

    PubMed Central

    Chappuis, François; Rijal, Suman; Soto, Alonso; Menten, Joris; Boelaert, Marleen

    2006-01-01

    Objective To compare the performance of the direct agglutination test and rK39 dipstick for the diagnosis of visceral leishmaniasis. Data sources Medline, citation tracking, January 1986 to December 2004. Selection criteria Original studies evaluating the direct agglutination test or the rK39 dipstick with clinical visceral leishmaniasis as target condition; adequate reference classification; and absolute numbers of true positive, true negative, false positive, and false negative observations available or derivable from the data presented. Results 30 studies evaluating the direct agglutination test and 13 studies evaluating the rK39 dipstick met the inclusion criteria. The combined sensitivity estimates of the direct agglutination test and the rK39 dipstick were 94.8% (95% confidence interval 92.7% to 96.4%) and 93.9% (87.7% to 97.1%), respectively. Sensitivity seemed higher and more homogenous in the studies carried out in South Asia. Specificity estimates were influenced by the type of controls. In phase III studies carried out on patients with clinically suspected disease, the estimated specificity of the direct agglutination test was 85.9% (72.3% to 93.4%) and of the rK39 dipstick was 90.6% (66.8% to 97.9%). Conclusion The diagnostic performance of the direct agglutination test and the rK39 dipstick for visceral leishmaniasis is good to excellent and seem comparable. PMID:16882683

  9. Regulation of Arabidopsis MAPKKK18 by ABI1 and SnRK2, components of the ABA signaling pathway.

    PubMed

    Tajdel, Małgorzata; Mituła, Filip; Ludwików, Agnieszka

    2016-04-01

    The plant hormone abscisic acid (ABA), a key regulator in many crucial developmental and physiological processes, recruits diverse components into precisely regulated signaling network. We recently discovered that MAPKKK18, an ABA-activated kinase, is regulated by the protein phosphatase type 2C (PP2C) ABI1 and the kinase SnRK2.6, both components of the ABA core signaling pathway. ABI1 acts to inhibit MAPKKK18 kinase activity, but also affects MAPKKK18 protein turnover via the ubiquitin-proteasome pathway. SnRK2.6 kinase also seems to be important for the regulation of MAPKKK18 function. In this review we summarize the mechanisms that are exclusively involved in MAPKKK18 kinase regulation and that ensure specificity in its activation. PMID:26852793

  10. The activity of SnRK1 is increased in Phaseolus vulgaris seeds in response to a reduced nutrient supply

    PubMed Central

    Coello, Patricia; Martínez-Barajas, Eleazar

    2014-01-01

    Phaseolus vulgaris seeds can grow and develop at the expense of the pod reserves after the fruits have been removed from the plant (Fountain etal., 1989). Because this process involves sensing the reduction of nutrients and the remobilisation of pod reserves, we investigated the effect on sucrose non-fermenting related kinase 1 (SnRK1) activity during this process. Bean fruits removed from the plant at 20 days after flowering (DAF) demonstrated active remobilisation of nutrients from the pod to the seeds. After 5 days, the pod dry weight was reduced by 50%. The process was characterized by a rapid degradation of starch, with the greatest decrease observed on day 1 after the fruits were removed. The pod nutrients were insufficient for the needs of all the seeds, and only some seeds continued their development. Those seeds exhibited a transient reduction in sucrose levels on day 1 after the fruits were removed. However, the normal level of sucrose was recovered, and the rate of starch synthesis was identical to that of a seed developed under normal conditions. Removing the fruits from the plant had no effect on the activity of SnRK1 in the pods, whereas in the seeds, the activity was increased by 35%. Simultaneously, a large reduction in seed sucrose levels was observed. The increase in SnRK1 activity was observed in both the cotyledon and embryo axes, but it was higher in the cotyledon. At 20–25 DAF, cotyledons actively accumulate storage materials. It is possible that the increase in SnRK1 activity observed in seeds developed in fruits that have been removed from the plant is part of the mechanism required for nutrient remobilisation under conditions of stress. PMID:24860586

  11. Functional analysis of duplicated Symbiosis Receptor Kinase (SymRK) genes during nodulation and mycorrhizal infection in soybean (Glycine max).

    PubMed

    Indrasumunar, Arief; Wilde, Julia; Hayashi, Satomi; Li, Dongxue; Gresshoff, Peter M

    2015-03-15

    Association between legumes and rhizobia results in the formation of root nodules, where symbiotic nitrogen fixation occurs. The early stages of this association involve a complex of signalling events between the host and microsymbiont. Several genes dealing with early signal transduction have been cloned, and one of them encodes the leucine-rich repeat (LRR) receptor kinase (SymRK; also termed NORK). The Symbiosis Receptor Kinase gene is required by legumes to establish a root endosymbiosis with Rhizobium bacteria as well as mycorrhizal fungi. Using degenerate primer and BAC sequencing, we cloned duplicated SymRK homeologues in soybean called GmSymRKα and GmSymRKβ. These duplicated genes have high similarity of nucleotide (96%) and amino acid sequence (95%). Sequence analysis predicted a malectin-like domain within the extracellular domain of both genes. Several putative cis-acting elements were found in promoter regions of GmSymRKα and GmSymRKβ, suggesting a participation in lateral root development, cell division and peribacteroid membrane formation. The mutant of SymRK genes is not available in soybean; therefore, to know the functions of these genes, RNA interference (RNAi) of these duplicated genes was performed. For this purpose, RNAi construct of each gene was generated and introduced into the soybean genome by Agrobacterium rhizogenes-mediated hairy root transformation. RNAi of GmSymRKβ gene resulted in an increased reduction of nodulation and mycorrhizal infection than RNAi of GmSymRKα, suggesting it has the major activity of the duplicated gene pair. The results from the important crop legume soybean confirm the joint phenotypic action of GmSymRK genes in both mycorrhizal and rhizobial infection seen in model legumes. PMID:25617765

  12. Measuring the levels of noise at the İstanbul Atatürk Airport and comparisons with model simulations.

    PubMed

    Sari, Deniz; Ozkurt, Nesimi; Akdag, Ali; Kutukoglu, Murat; Gurarslan, Aliye

    2014-06-01

    Airport noise and its impact on the surrounding areas are major issues in the aviation industry. The İstanbul Atatürk Airport is a major global airport with passenger numbers increasing rapidly per annum. The noise levels for day, evening and night times were modeled around the İstanbul Atatürk Airport according to the European Noise Directive using the actual data records for the year 2011. The "ECAC Doc. 29-Interim" method was used for the computation of the aircraft traffic noise. In the setting the noise model for the local airport topography was taken into consideration together with the noise source data, the airport loadings, features of aircraft and actual air traffic data. Model results were compared with long-term noise measurement values for calibration. According to calibration results, classifications of the aircraft type and flight tracks were revised. For noise model validation, the daily noise measurements at four additional locations were used during the verification period. The input data was re-edited only for these periods and the model was validated. A successful model performance was obtained in several zones around the airport. The validated noise model of the İstanbul Atatürk Airport can be now utilized both for determining the noise levels in the future and for producing new strategies which are about the land use planning, operational considerations for the air traffic management and the noise abatement procedures. PMID:23972900

  13. Characterization of anti-Legionella activity of warnericin RK and delta-lysin I from Staphylococcus warneri.

    PubMed

    Verdon, Julien; Berjeaud, Jean-Marc; Lacombe, Christian; Héchard, Yann

    2008-06-01

    Legionella pneumophila, the causative agent of Legionnaires' disease, is a waterborne bacteria. It can multiply in man-made water systems and infect people who inhale contaminated droplets. We have previously reported a Staphylococcus warneri strain that display an anti-Legionella activity. In this work, we characterized three anti-Legionella peptides that are produced by S. warneri. One peptide, warnericin RK, is original, while the two others are delta-lysin I and delta-lysin II, whose genes were previously described. Due to high sequence similarity of the two delta-lysins, further characterization was performed only on delta-lysin I. Warnericin RK and delta-lysin I displayed the same antibacterial spectrum, which is almost restricted to the Legionella genus. Also, both peptides have a hemolytic activity. These results led to the hypothesis that warnericin RK and delta-lysin I share a similar mode of action, and that Legionella should have a specific feature that may explain the high specificity of these antibacterial peptides. PMID:18339450

  14. MLK-3 activates the SAPK/JNK and p38/RK pathways via SEK1 and MKK3/6.

    PubMed Central

    Tibbles, L A; Ing, Y L; Kiefer, F; Chan, J; Iscove, N; Woodgett, J R; Lassam, N J

    1996-01-01

    Mixed lineage kinase-3 (MLK-3) is a 97 kDa serine/threonine kinase with multiple interaction domains, including a Cdc42 binding motif, but unknown function. Cdc42 and the related small GTP binding protein Rac1 can activate the SAPK/JNK and p38/RK stress-responsive kinase cascades, suggesting that MLK-3 may have a role in upstream regulation of these pathways. In support of this role, we demonstrate that MLK-3 can specifically activate the SAPK/JNK and p38/RK pathways, but has no effect on the activation of ERKs. Immunoprecipitated MLK-3 catalyzed the phosphorylation of SEK1 in vitro, and co-transfected MLK-3 induced phosphorylation of SEK1 and MKK3 at sites required for activation, suggesting direct regulation of these protein kinases. Furthermore, interactions between MLK-3 and SEK and MLK-3 and MKK6 were observed in co-precipitation experiments. Finally, kinase-dead mutants of MLK-3 blocked activation of the SAPK pathway by a newly identified mammalian analog of Ste20, germinal center kinase, but not by MEKK, suggesting that MLK-3 functions to activate the SAPK/JNK and p38/RK cascades in response to stimuli transduced by Ste20-like kinases. Images PMID:9003778

  15. An Initial Investigation of the Effects of Turbulence Models on the Convergence of the RK/Implicit Scheme

    NASA Technical Reports Server (NTRS)

    Swanson, R. C.; Rossow, C.-C.

    2008-01-01

    A three-stage Runge-Kutta (RK) scheme with multigrid and an implicit preconditioner has been shown to be an effective solver for the fluid dynamic equations. This scheme has been applied to both the compressible and essentially incompressible Reynolds-averaged Navier-Stokes (RANS) equations using the algebraic turbulence model of Baldwin and Lomax (BL). In this paper we focus on the convergence of the RK/implicit scheme when the effects of turbulence are represented by either the Spalart-Allmaras model or the Wilcox k-! model, which are frequently used models in practical fluid dynamic applications. Convergence behavior of the scheme with these turbulence models and the BL model are directly compared. For this initial investigation we solve the flow equations and the partial differential equations of the turbulence models indirectly coupled. With this approach we examine the convergence behavior of each system. Both point and line symmetric Gauss-Seidel are considered for approximating the inverse of the implicit operator of the flow solver. To solve the turbulence equations we use a diagonally dominant alternating direction implicit (DDADI) scheme. Computational results are presented for three airfoil flow cases and comparisons are made with experimental data. We demonstrate that the two-dimensional RANS equations and transport-type equations for turbulence modeling can be efficiently solved with an indirectly coupled algorithm that uses the RK/implicit scheme for the flow equations.

  16. N-Myristoylation Regulates the SnRK1 Pathway in Arabidopsis[W

    PubMed Central

    Pierre, Michèle; Traverso, José A.; Boisson, Bertrand; Domenichini, Séverine; Bouchez, David; Giglione, Carmela; Meinnel, Thierry

    2007-01-01

    Cotranslational and posttranslational modifications are increasingly recognized as important in the regulation of numerous essential cellular functions. N-myristoylation is a lipid modification ensuring the proper function and intracellular trafficking of proteins involved in many signaling pathways. Arabidopsis thaliana, like human, has two tightly regulated N-myristoyltransferase (NMT) genes, NMT1 and NMT2. Characterization of knockout mutants showed that NMT1 was strictly required for plant viability, whereas NMT2 accelerated flowering. NMT1 impairment induced extremely severe defects in the shoot apical meristem during embryonic development, causing growth arrest after germination. A transgenic plant line with an inducible NMT1 gene demonstrated that NMT1 expression had further effects at later stages. NMT2 did not compensate for NMT1 in the nmt1-1 mutant, but NMT2 overexpression resulted in shoot and root meristem abnormalities. Various data from complementation experiments in the nmt1-1 background, using either yeast or human NMTs, demonstrated a functional link between the developmental arrest of nmt1-1 mutants and the myristoylation state of an extremely small set of protein targets. We show here that protein N-myristoylation is systematically associated with shoot meristem development and that SnRK1 (for SNF1-related kinase) is one of its essential primary targets. PMID:17827350

  17. Assessment of mitral Björk-Shiley prosthetic dysfunction using digitised M mode echocardiography.

    PubMed Central

    Dawkins, K D; Cotter, L; Gibson, D G

    1984-01-01

    Digitised M mode echocardiograms were analysed in 22 patients with possible Björk-Shiley mitral prosthetic dysfunction. Patients with paraprosthetic mitral regurgitation had a significantly greater shortening fraction, an increased peak rate of dimension change during systole, and an increased peak velocity of circumferential fibre shortening than those with poor left ventricular function. Patients with a clotted prosthesis had lower values for shortening fraction and peak rate of dimension change during systole than patients with paraprosthetic regurgitation. In this latter group, the peak rate of dimension change during diastole and peak lengthening rate were greater than in either those patients with poor left ventricular function or those with a clotted prosthesis. In addition, the peak lengthening rate was greater in those with a clotted prosthesis than in those with poor left ventricular function. Thus M mode echocardiography is a useful method of assessing mitral prosthetic dysfunction and allows patients with paraprosthetic regurgitation to be distinguished from those with either poor left ventricular function or a clotted prosthesis. PMID:6691866

  18. The β-Subunit of the SnRK1 Complex Is Phosphorylated by the Plant Cell Death Suppressor Adi31[C][W][OA

    PubMed Central

    Avila, Julian; Gregory, Oliver G.; Su, Dongyin; Deeter, Taunya A.; Chen, Sixue; Silva-Sanchez, Cecilia; Xu, Shouling; Martin, Gregory B.; Devarenne, Timothy P.

    2012-01-01

    The protein kinase AvrPto-dependent Pto-interacting protein3 (Adi3) is a known suppressor of cell death, and loss of its function has been correlated with cell death induction during the tomato (Solanum lycopersicum) resistance response to its pathogen Pseudomonas syringae pv tomato. However, Adi3 downstream interactors that may play a role in cell death regulation have not been identified. We used a yeast two-hybrid screen to identify the plant SnRK1 (for Sucrose non-Fermenting-1-Related Protein Kinase1) protein as an Adi3-interacting protein. SnRK1 functions as a regulator of carbon metabolism and responses to biotic and abiotic stresses. SnRK1 exists in a heterotrimeric complex with a catalytic α-subunit (SnRK1), a substrate-interacting β-subunit, and a regulatory γ-subunit. Here, we show that Adi3 interacts with, but does not phosphorylate, the SnRK1 α-subunit. The ability of Adi3 to phosphorylate the four identified tomato β-subunits was also examined, and it was found that only the Galactose Metabolism83 (Gal83) β-subunit was phosphorylated by Adi3. This phosphorylation site on Gal83 was identified as serine-26 using a mutational approach and mass spectrometry. In vivo expression of Gal83 indicates that it contains multiple phosphorylation sites, one of which is serine-26. An active SnRK1 complex containing Gal83 as the β-subunit and sucrose nonfermenting4 as the γ-subunit was constructed to examine functional aspects of the Adi3 interaction with SnRK1 and Gal83. These assays revealed that Adi3 is capable of suppressing the kinase activity of the SnRK1 complex through Gal83 phosphorylation plus the interaction with SnRK1 and suggested that this function may be related to the cell death suppression activity of Adi3. PMID:22573803

  19. Two-Component System VicRK Regulates Functions Associated with Establishment of Streptococcus sanguinis in Biofilms

    PubMed Central

    Moraes, Julianna J.; Stipp, Rafael N.; Harth-Chu, Erika N.; Camargo, Tarsila M.; Höfling, José F.

    2014-01-01

    Streptococcus sanguinis is a commensal pioneer colonizer of teeth and an opportunistic pathogen of infectious endocarditis. The establishment of S. sanguinis in host sites likely requires dynamic fitting of the cell wall in response to local stimuli. In this study, we investigated the two-component system (TCS) VicRK in S. sanguinis (VicRKSs), which regulates genes of cell wall biogenesis, biofilm formation, and virulence in opportunistic pathogens. A vicK knockout mutant obtained from strain SK36 (SKvic) showed slight reductions in aerobic growth and resistance to oxidative stress but an impaired ability to form biofilms, a phenotype restored in the complemented mutant. The biofilm-defective phenotype was associated with reduced amounts of extracellular DNA during aerobic growth, with reduced production of H2O2, a metabolic product associated with DNA release, and with inhibitory capacity of S. sanguinis competitor species. No changes in autolysis or cell surface hydrophobicity were detected in SKvic. Reverse transcription-quantitative PCR (RT-qPCR), electrophoretic mobility shift assays (EMSA), and promoter sequence analyses revealed that VicR directly regulates genes encoding murein hydrolases (SSA_0094, cwdP, and gbpB) and spxB, which encodes pyruvate oxidase for H2O2 production. Genes previously associated with spxB expression (spxR, ccpA, ackA, and tpK) were not transcriptionally affected in SKvic. RT-qPCR analyses of S. sanguinis biofilm cells further showed upregulation of VicRK targets (spxB, gbpB, and SSA_0094) and other genes for biofilm formation (gtfP and comE) compared to expression in planktonic cells. This study provides evidence that VicRKSs regulates functions crucial for S. sanguinis establishment in biofilms and identifies novel VicRK targets potentially involved in hydrolytic activities of the cell wall required for these functions. PMID:25183732

  20. Cloning and characterization of a maize SnRK2 protein kinase gene confers enhanced salt tolerance in transgenic Arabidopsis.

    PubMed

    Ying, Sheng; Zhang, Deng-Feng; Li, Hui-Yong; Liu, Ying-Hui; Shi, Yun-Su; Song, Yan-Chun; Wang, Tian-Yu; Li, Yu

    2011-09-01

    SnRK2 (sucrose non-fermenting 1-related protein kinases 2) represents a unique family of protein kinase in regulating signaling transduction in plants. Although the regulatory mechanisms of SnRK2 have been well demonstrated in Arabidopsis thaliana, their functions in maize are still unknown. In our study, we cloned an SnRK2 gene from maize, ZmSAPK8, which encoded a putative homolog of the rice SAPK8 protein. ZmSAPK8 had two copies in the maize genome and harbored eight introns in its coding region. We demonstrated that ZmSAPK8 expressed differentially in various organs of maize plants and was up-regulated by high-salinity and drought treatment. A green fluorescent protein (GFP)-tagged ZmSAPK8 showed subcellular localization in the cell membrane, cytoplasm and nucleus. In vitro kinase assays indicated that ZmSAPK8 preferred Mn(2+) to Mg(2+) as cofactor for phosphorylation, and Ser-182 and Thr-183 in activation loop was important for its activity. Heterologous overexpression of ZmSAPK8 in Arabidopsis could significantly strengthen tolerance to salt stress. Under salt treatment, ZmSAPK8-overexpressed transgenic plants exhibited higher germination rate and proline content, low electrolyte leakage and higher survival rate than wild type. Further analysis indicated that transgenic plants showed increased transcription of the stress-related genes, RD29A, RD29B, RAB18, ABI1, DREB2A and P5CS1, under high-salinity conditions. The results demonstrated that ZmSAPK8 was involved in diverse stress signal transduction. Moreover, no obvious adverse effects on growth and development in the ZmSAPK8-overexpressed transgenic plants implied that ZmSAPK8 was potentially useful in transgenic breeding to improve salt tolerance in crops. PMID:21638061

  1. Comparative proteomic analysis of early salt stress-responsive proteins in roots of SnRK2 transgenic rice

    PubMed Central

    2012-01-01

    Background The rice roots are highly salt-sensitive organ and primary root growth is rapidly suppressed by salt stress. Sucrose nonfermenting 1-related protein kinase2 (SnRK2) family is one of the key regulator of hyper-osmotic stress signalling in various plant cells. To understand early salt response of rice roots and identify SnRK2 signaling components, proteome changes of transgenic rice roots over-expressing OSRK1, a rice SnRK2 kinase were investigated. Results Proteomes were analyzed by two-dimensional electrophoresis and protein spots were identified by LC-MS/MS from wild type and OSRK1 transgenic rice roots exposed to 150 mM NaCl for either 3 h or 7 h. Fifty two early salt -responsive protein spots were identified from wild type rice roots. The major up-regulated proteins were enzymes related to energy regulation, amino acid metabolism, methylglyoxal detoxification, redox regulation and protein turnover. It is noted that enzymes known to be involved in GA-induced root growth such as fructose bisphosphate aldolase and methylmalonate semialdehyde dehydrogenase were clearly down-regulated. In contrast to wild type rice roots, only a few proteins were changed by salt stress in OSRK1 transgenic rice roots. A comparative quantitative analysis of the proteome level indicated that forty three early salt-responsive proteins were magnified in transgenic rice roots at unstressed condition. These proteins contain single or multiple potential SnRK2 recognition motives. In vitro kinase assay revealed that one of the identified proteome, calreticulin is a good substrate of OSRK1. Conclusions Our present data implicate that rice roots rapidly changed broad spectrum of energy metabolism upon challenging salt stress, and suppression of GA signaling by salt stress may be responsible for the rapid arrest of root growth and development. The broad spectrum of functional categories of proteins affected by over-expression of OSRK1 indicates that OSRK1 is an upstream regulator of

  2. The sucrose non-fermenting-1-related (SnRK) family of protein kinases: potential for manipulation to improve stress tolerance and increase yield.

    PubMed

    Coello, Patricia; Hey, Sandra J; Halford, Nigel G

    2011-01-01

    Sucrose non-fermenting-1 (SNF1)-related protein kinases (SnRKs) take their name from their fungal homologue, SNF1, a global regulator of carbon metabolism. The plant family has burgeoned to comprise 38 members which can be subdivided into three sub-families: SnRK1, SnRK2, and SnRK3. There is now good evidence that this has occurred to allow plants to link metabolic and stress signalling in a way that does not occur in other organisms. The role of SnRKs, focusing in particular on abscisic acid-induced signalling pathways, salinity tolerance, responses to nutritional stress and disease, and the regulation of carbon metabolism and, therefore, yield, is reviewed here. The key role that SnRKs play at the interface between metabolic and stress signalling make them potential candidates for manipulation to improve crop performance in extreme environments. PMID:20974737

  3. The kilE locus of promiscuous IncP alpha plasmid RK2 is required for stable maintenance in Pseudomonas aeruginosa.

    PubMed Central

    Wilson, J W; Sia, E A; Figurski, D H

    1997-01-01

    Eight coordinately regulated operons constitute the kor regulon of the IncP alpha plasmid RK2. Three operons specify functions required for replication initiation, conjugative transfer, and control of gene expression. The functions of the other operons, including those of the four coregulated operons that compose the kilA, kilC, and kilE loci, have not been determined. Here, we present the first evidence that a kil determinant is involved in IncP plasmid maintenance. Elevation of KorC levels specifically to reduce the expression of the KorC-regulated kilC and kilE operons severely affected the maintenance of both the IncP alpha plasmid RK2lac and the IncP beta plasmid R751 in Pseudomonas aeruginosa but had little effect on plasmid maintenance in Escherichia coli. Precise deletion of the two kilE operons from RK2lac was achieved with the VEX mutagenesis system for large genomes. The resulting plasmid showed significant loss of stability in P. aeruginosa only. The defect could be complemented by reintroduction of kilE at a different position on the plasmid. The instability of the RK2lac delta kilE mutant did not result from a reduction in average plasmid copy number, reduced expression of kilC, decreased conjugative transfer, or loss of the korE regulator. We found that both the par and kilE loci are required for full stability of RK2lac in P. aeruginosa and that the par and kilE functions act independently. These results demonstrate a critical role for the kilE locus in the stable inheritance of RK2 in P. aeruginosa. PMID:9079921

  4. Evaluation of Chicken IgY Generated Against Canine Parvovirus Viral-Like Particles and Development of Enzyme-Linked Immunosorbent Assay and Immunochromatographic Assay for Canine Parvovirus Detection.

    PubMed

    He, Jinxin; Wang, Yuan; Sun, Shiqi; Zhang, Xiaoying

    2015-11-01

    Immunoglobulin Y (IgY) antibodies were generated against canine parvovirus virus-like particles (CPV-VLPs) antigen using chickens. Anti-CPV-VLPs-IgY was extracted from hen egg yolk and used for developing enzyme-linked immunosorbent assay (ELISA) and immunochromatographic assay (ICA) for the detection of CPV in dog feces. The cutoff negative values for anti-CPV-VLPs-IgY were determined using negative fecal samples (already confirmed by polymerase chain reaction [PCR]). In both ELISA and ICA, there was no cross-reaction with other diarrheal pathogens. Thirty-four fecal samples were collected from dogs with diarrhea, of which 26.47% were confirmed as CPV-positive samples by PCR, while 29.41% and 32.35% of the samples were found to be positive by ELISA and ICA, respectively. The developed ELISA and ICA exhibited 97.06% and 94.12% conformity with PCR. Higher sensitivity and specificity were observed for IgY-based ELISA and ICA. Thus, they could be suitable for routine use in the diagnosis of CPV in dogs. PMID:26469376

  5. Modeling of noise pollution and estimated human exposure around İstanbul Atatürk Airport in Turkey.

    PubMed

    Ozkurt, Nesimi; Sari, Deniz; Akdag, Ali; Kutukoglu, Murat; Gurarslan, Aliye

    2014-06-01

    The level of aircraft noise exposure around İstanbul Atatürk Airport was calculated according to the European Noise Directive. These calculations were based on the actual flight data for each flight in the year 2011. The study area was selected to cover of 25km radius centered on the Aerodrome Reference Point of the airport. The geographical data around İstanbul Atatürk Airport was used to prepare elevation, residential building, auxiliary building, hospital and school layers in SoundPlan software. It was found that 1.2% of the land area of İstanbul City exceeds the threshold of 55dB(A) during daytime. However, when the exceedance of threshold of 65dB(A)is investigated, the affected area is found quite small (0.2% of land area of city). About 0.3% of the land area of İstanbul City has noise levels exceeding 55dB(A) during night-time. Our results show that about 4% of the resident population was exposed to 55dB(A) or higher noises during daytime in İstanbul. When applying the second threshhold criteria, nearly 1% of the population is exposed to noise levels greater than 65dB(A). At night-time, 1.3% of the population is exposed to 55dB(A) or higher noise levels. PMID:23998505

  6. An rK28-Based Immunoenzymatic Assay for the Diagnosis of Canine Visceral Leishmaniasis in Latin America.

    PubMed

    Lauricella, Marta Alicia; Maidana, Cristina Graciela; Frias, Victoria Fragueiro; Romagosa, Carlo M; Negri, Vanesa; Benedetti, Ruben; Sinagra, Angel J; Luna, Concepcion; Tartaglino, Lilian; Laucella, Susana; Reed, Steven G; Riarte, Adelina R

    2016-07-01

    Direct observation of Leishmania parasites in tissue aspirates has shown low sensitivity for the detection of canine visceral leishmaniasis (VL). Therefore in the last quarter century immunoenzymatic tests have been developed to improve diagnosis. The purpose of this study was to develop a fast recombinant K28 antigen, naked-eye qualitative enzyme-linked immunosorbent assay (VL Ql-ELISA) and a quantitative version (VL Qt-ELISA), and to display it in a kit format, whose cutoff value (0.156) was selected as the most adequate one to differentiate reactive from nonreactive samples. Considering 167 cases and 300 controls, sensitivity was 91% for both assays and specificity was 100% and 98.7% in Ql-ELISA and Qt-ELISA, respectively. Positive predictive values were 100% and 97.4% for Ql-ELISA and Qt-ELISA, respectively, and negative predictive values were 95.2% for both ELISAs. Reagent stability, reliability studies, including periodic repetitions and retest of samples, cutoff selection, and comparison of rK28 ELISAs with rK39 immunochromatographic test, were the international criteria that supported the quality in both kits. The performance of both ELISA kits in this work confirmed their validity and emphasized their usefulness for low-to-medium complexity laboratories. PMID:27162270

  7. Effects of flavorings, storage conditions, and storage time on survival of Staphylococcus aureus in Sürk cheese.

    PubMed

    Masatcioğlu, Tuğrul M; Avşar, Yahya K

    2005-07-01

    The objectives of this study were to determine the cumulative effects of flavorings (chili pepper, thyme, mint, cumin, nutmeg, allspice, clove, cinnamon, black pepper, salt, and hot red pepper paste), storage conditions, and storage time on the survival of Staphylococcus aureus in Sürk cheese and to monitor the associated chemical changes. Sürk cheese, a traditional Turkish cheese, was produced by heating diluted nonfat yogurt and adding flavorings to the resultant acid-heat curd. The cheese was later inoculated with S. aureus, shaped conically, and stored aerobically for mold growth and anaerobically in olive oil for 30 days at room temperature. The moisture content of aerobically stored cheese decreased over time and led to increases in total solids, salt, salt-in-moisture, and ash content during ripening (P < 0.05). The presence or absence of the flavorings had no significant effect, whereas storage conditions and storage duration decreased the survival of S. aureus (P < 0.05). PMID:16013393

  8. Vancomycin susceptibility in methicillin-resistant Staphylococcus aureus is mediated by YycHI activation of the WalRK essential two-component regulatory system

    PubMed Central

    Cameron, David R.; Jiang, Jhih-Hang; Kostoulias, Xenia; Foxwell, Daniel J.; Peleg, Anton Y.

    2016-01-01

    The treatment of infections caused by methicillin-resistant Staphylococcus aureus is complicated by the emergence of strains with intermediate-level resistance to vancomycin (termed VISA). We have characterised a molecular pathway involved in the in vivo evolution of VISA mediated by the regulatory proteins YycH and YycI. In contrast to their function in other bacterial species, we report a positive role for these auxiliary proteins in regulation of the two-component regulator WalRK. Transcriptional profiling of yycH and yycI deletion mutants revealed downregulation of the ‘WalRK regulon’ including cell wall hydrolase genes atlA and sle1, with functional autolysis assays supporting these data by showing an impaired autolytic phenotype for each deletion strain. Using bacterial-two hybrid assays, we showed that YycH and YycI interact, and that YycHI also interacts with the sensor kinase WalK, forming a ternary protein complex. Mutation to YycH or YycI associated with clinical VISA strains had a deleterious impact on the YycHI/WalK complex, suggesting that the interaction is important for the regulation of WalRK. Taken together, we have described a novel antibiotic resistance strategy for the human pathogen S. aureus, whereby YycHI mutations are selected for in vivo leading to reduced WalRK activation, impaired cell wall turnover and ultimately reduced vancomycin efficacy. PMID:27600558

  9. Vancomycin susceptibility in methicillin-resistant Staphylococcus aureus is mediated by YycHI activation of the WalRK essential two-component regulatory system.

    PubMed

    Cameron, David R; Jiang, Jhih-Hang; Kostoulias, Xenia; Foxwell, Daniel J; Peleg, Anton Y

    2016-01-01

    The treatment of infections caused by methicillin-resistant Staphylococcus aureus is complicated by the emergence of strains with intermediate-level resistance to vancomycin (termed VISA). We have characterised a molecular pathway involved in the in vivo evolution of VISA mediated by the regulatory proteins YycH and YycI. In contrast to their function in other bacterial species, we report a positive role for these auxiliary proteins in regulation of the two-component regulator WalRK. Transcriptional profiling of yycH and yycI deletion mutants revealed downregulation of the 'WalRK regulon' including cell wall hydrolase genes atlA and sle1, with functional autolysis assays supporting these data by showing an impaired autolytic phenotype for each deletion strain. Using bacterial-two hybrid assays, we showed that YycH and YycI interact, and that YycHI also interacts with the sensor kinase WalK, forming a ternary protein complex. Mutation to YycH or YycI associated with clinical VISA strains had a deleterious impact on the YycHI/WalK complex, suggesting that the interaction is important for the regulation of WalRK. Taken together, we have described a novel antibiotic resistance strategy for the human pathogen S. aureus, whereby YycHI mutations are selected for in vivo leading to reduced WalRK activation, impaired cell wall turnover and ultimately reduced vancomycin efficacy. PMID:27600558

  10. The replication initiation protein of the broad-host-range plasmid RK2 is activated by the ClpX chaperone.

    PubMed

    Konieczny, I; Helinski, D R

    1997-12-23

    Initiation and control of replication of the broad-host-range plasmid RK2 requires two plasmid-encoded elements, the replication origin (oriV) and the initiation protein TrfA. Purified TrfA is largely in the form of a dimer; however, only the monomeric form of the protein can bind specifically to the direct repeats (iterons) at the RK2 origin. The largely dimeric form of wild-type TrfA is inactive in the initiation of replication of RK2 in an in vitro replication system reconstituted from purified components. However, preincubation of the TrfA protein with the ClpX molecular chaperone isolated from Escherichia coli activates the initiator protein for replication in the purified system. We further observed that ClpX, in an ATP-dependent reaction, greatly increases the proportion of TrfA monomers and, therefore, the ability of this protein to bind to iterons localized within RK2 origin. Finally, a copy-up mutant of the TrfA protein which is largely in the monomer form is active in the reconstituted in vitro replication system, and its activity is not affected by ClpX. PMID:9405620

  11. CovR and VicRK regulate cell surface biogenesis genes required for biofilm formation in Streptococcus mutans.

    PubMed

    Stipp, Rafael N; Boisvert, Heike; Smith, Daniel J; Höfling, José F; Duncan, Margaret J; Mattos-Graner, Renata O

    2013-01-01

    The two-component system VicRK and the orphan regulator CovR of Streptococcus mutans co-regulate a group of virulence genes associated with the synthesis of and interaction with extracellular polysaccharides of the biofilm matrix. Knockout mutants of vicK and covR display abnormal cell division and morphology phenotypes, although the gene function defects involved are as yet unknown. Using transcriptomic comparisons between parent strain UA159 with vicK (UAvic) or covR (UAcov) deletion mutants together with electrophoretic motility shift assays (EMSA), we identified genes directly regulated by both VicR and CovR with putative functions in cell wall/surface biogenesis, including gbpB, wapE, smaA, SMU.2146c, and lysM. Deletion mutants of genes regulated by VicR and CovR (wapE, lysM, smaA), or regulated only by VicR (SMU.2146c) or CovR (epsC) promoted significant alterations in biofilm initiation, including increased fragility, defects in microcolony formation, and atypical cell morphology and/or chaining. Significant reductions in mureinolytic activity and/or increases in DNA release during growth were observed in knockout mutants of smaA, wapE, lysM, SMU.2146c and epsC, implying roles in cell wall biogenesis. WapE and lysM mutations also affected cell hydrophobicity and sensitivity to osmotic or oxidative stress. Finally, vicR, covR and VicRK/CovR-targets (gbpB, wapE, smaA, SMU.2146c, lysM, epsC) are up-regulated in UA159 during biofilm initiation, in a sucrose-dependent manner. These data support a model in which VicRK and CovR coordinate cell division and surface biogenesis with the extracellular synthesis of polysaccharides, a process apparently required for formation of structurally stable biofilms in the presence of sucrose. PMID:23554881

  12. CovR and VicRK Regulate Cell Surface Biogenesis Genes Required for Biofilm Formation in Streptococcus mutans

    PubMed Central

    Stipp, Rafael N.; Boisvert, Heike; Smith, Daniel J.; Höfling, José F.; Duncan, Margaret J.; Mattos-Graner, Renata O.

    2013-01-01

    The two-component system VicRK and the orphan regulator CovR of Streptococcus mutans co-regulate a group of virulence genes associated with the synthesis of and interaction with extracellular polysaccharides of the biofilm matrix. Knockout mutants of vicK and covR display abnormal cell division and morphology phenotypes, although the gene function defects involved are as yet unknown. Using transcriptomic comparisons between parent strain UA159 with vicK (UAvic) or covR (UAcov) deletion mutants together with electrophoretic motility shift assays (EMSA), we identified genes directly regulated by both VicR and CovR with putative functions in cell wall/surface biogenesis, including gbpB, wapE, smaA, SMU.2146c, and lysM. Deletion mutants of genes regulated by VicR and CovR (wapE, lysM, smaA), or regulated only by VicR (SMU.2146c) or CovR (epsC) promoted significant alterations in biofilm initiation, including increased fragility, defects in microcolony formation, and atypical cell morphology and/or chaining. Significant reductions in mureinolytic activity and/or increases in DNA release during growth were observed in knockout mutants of smaA, wapE, lysM, SMU.2146c and epsC, implying roles in cell wall biogenesis. WapE and lysM mutations also affected cell hydrophobicity and sensitivity to osmotic or oxidative stress. Finally, vicR, covR and VicRK/CovR-targets (gbpB, wapE, smaA, SMU.2146c, lysM, epsC) are up-regulated in UA159 during biofilm initiation, in a sucrose-dependent manner. These data support a model in which VicRK and CovR coordinate cell division and surface biogenesis with the extracellular synthesis of polysaccharides, a process apparently required for formation of structurally stable biofilms in the presence of sucrose. PMID:23554881

  13. Phosphoproteomic identification of targets of the Arabidopsis sucrose nonfermenting-like kinase SnRK2.8 reveals a connection to metabolic processes

    PubMed Central

    Shin, Ryoung; Alvarez, Sophie; Burch, Adrien Y.; Jez, Joseph M.; Schachtman, Daniel P.

    2007-01-01

    SnRK2.8 is a member of the sucrose nonfermenting-related kinase family that is down-regulated when plants are deprived of nutrients and growth is reduced. When this kinase is over expressed in Arabidopsis, the plants grow larger. To understand how this kinase modulates growth, we identified some of the proteins that are phosphorylated by this kinase. A new phosphoproteomic method was used in which total protein from plants overexpressing the kinase was compared with total protein from plants in which the kinase was inactivated. Protein profiles were compared on two-dimensional gels following staining by a dye that recognizes phosphorylated amino acids. Candidate target proteins were confirmed with in vitro phosphorylation assays, using the kinase and target proteins that were purified from Escherichia coli. Seven target proteins were confirmed as being phosphorylated by SnRK2.8. Certain targets, such as 14-3-3 proteins, regulate as yet unidentified proteins, whereas other targets, such as glyoxalase I and ribose 5-phosphate isomerase, detoxify byproducts from glycolysis and catalyze one of the final steps in carbon fixation, respectively. Also, adenosine kinase and 60S ribosomal protein were confirmed as targets of SnRK2.8. Using mass spectrometry, we identified phosphorylated residues in the SnRK2.8, the 14-3-3κ, and the 14-3-3χ. These data show that the expression of SnRK2.8 is correlated with plant growth, which may in part be due to the phosphorylation of enzymes involved in metabolic processes. PMID:17404219

  14. Four Arabidopsis AREB/ABF transcription factors function predominantly in gene expression downstream of SnRK2 kinases in abscisic acid signalling in response to osmotic stress.

    PubMed

    Yoshida, Takuya; Fujita, Yasunari; Maruyama, Kyonoshin; Mogami, Junro; Todaka, Daisuke; Shinozaki, Kazuo; Yamaguchi-Shinozaki, Kazuko

    2015-01-01

    Under osmotic stress conditions such as drought and high salinity, the plant hormone abscisic acid (ABA) plays important roles in stress-responsive gene expression mainly through three bZIP transcription factors, AREB1/ABF2, AREB2/ABF4 and ABF3, which are activated by SNF1-related kinase 2s (SnRK2s) such as SRK2D/SnRK2.2, SRK2E/SnRK2.6 and SRK2I/SnRK2.3 (SRK2D/E/I). However, since the three AREB/ABFs are crucial, but not exclusive, for the SnRK2-mediated gene expression, transcriptional pathways governed by SRK2D/E/I are not fully understood. Here, we show that a bZIP transcription factor, ABF1, is a functional homolog of AREB1, AREB2 and ABF3 in ABA-dependent gene expression in Arabidopsis. Despite lower expression levels of ABF1 than those of the three AREB/ABFs, the areb1 areb2 abf3 abf1 mutant plants displayed increased sensitivity to drought and decreased sensitivity to ABA in primary root growth compared with the areb1 areb2 abf3 mutant. Genome-wide transcriptome analyses revealed that expression of downstream genes of SRK2D/E/I, which include many genes functioning in osmotic stress responses and tolerance such as transcription factors and LEA proteins, was mostly impaired in the quadruple mutant. Thus, these results indicate that the four AREB/ABFs are the predominant transcription factors downstream of SRK2D/E/I in ABA signalling in response to osmotic stress during vegetative growth. PMID:24738645

  15. The hybrid four-CBS-domain KINβγ subunit functions as the canonical γ subunit of the plant energy sensor SnRK1.

    PubMed

    Ramon, Matthew; Ruelens, Philip; Li, Yi; Sheen, Jen; Geuten, Koen; Rolland, Filip

    2013-07-01

    The AMPK/SNF1/SnRK1 protein kinases are a family of ancient and highly conserved eukaryotic energy sensors that function as heterotrimeric complexes. These typically comprise catalytic α subunits and regulatory β and γ subunits, the latter function as the energy-sensing modules of animal AMPK through adenosine nucleotide binding. The ability to monitor accurately and adapt to changing environmental conditions and energy supply is essential for optimal plant growth and survival, but mechanistic insight in the plant SnRK1 function is still limited. In addition to a family of γ-like proteins, plants also encode a hybrid βγ protein that combines the Four-Cystathionine β-synthase (CBS)-domain (FCD) structure in γ subunits with a glycogen-binding domain (GBD), typically found in β subunits. We used integrated functional analyses by ectopic SnRK1 complex reconstitution, yeast mutant complementation, in-depth phylogenetic reconstruction, and a seedling starvation assay to show that only the hybrid KINβγ protein that recruited the GBD around the emergence of the green chloroplast-containing plants, acts as the canonical γ subunit required for heterotrimeric complex formation. Mutagenesis and truncation analysis further show that complex interaction in plant cells and γ subunit function in yeast depend on both a highly conserved FCD and a pre-CBS domain, but not the GBD. In addition to novel insight into canonical AMPK/SNF/SnRK1 γ subunit function, regulation and evolution, we provide a new classification of plant FCD genes as a convenient and reliable tool to predict regulatory partners for the SnRK1 energy sensor and novel FCD gene functions. PMID:23551663

  16. Perivalvular leakage 25 years after initial mitral valve replacement with a Björk-Shiley prosthesis.

    PubMed

    Minami, Hiroya; Asada, Tatsuro; Gan, Kunio

    2008-09-01

    An 80-year-old woman had undergone initial mitral valve replacement using a Björk-Shiley mechanical valve owing to mitral stenosis 25 years earlier. Suddenly, she had anemia and an increased lactic dehydrogenase (LDH) level. Transesophageal echography (TEE) showed perivalvular leakage. In a redo operation, two side-by-side stitches of the valve on the posterior annulus were loosened without cutting and the sewing cuff at that site was floated over the annulus, leading to the perivalvular leakage. The valve was easily removed; and round, hard, degenerative calcified tissue composed of remnant mitral valve in the suture site during the initial operation was found just under the sewing cuff. After resection of this calcified round tissue, a 25-mm bioprosthesis was put in place. Her postoperative recovery was uneventful, and 47 days after surgery she was discharged without perivalvular leakage or anemia. PMID:18791673

  17. Trehalose-6-phosphate and SnRK1 kinases in plant development and signaling: the emerging picture

    PubMed Central

    Tsai, Allen Y.-L.; Gazzarrini, Sonia

    2014-01-01

    Carbohydrates, or sugars, regulate various aspects of plant growth through modulation of cell division and expansion. Besides playing essential roles as sources of energy for growth and as structural components of cells, carbohydrates also regulate the timing of expression of developmental programs. The disaccharide trehalose is used as an energy source, as a storage and transport molecule for glucose, and as a stress-responsive compound important for cellular protection during stress in all kingdoms. Trehalose, however, is found in very low amounts in most plants, pointing to a signaling over metabolic role for this non-reducing disaccharide. In the last decade, trehalose-6-phosphate (T6P), an intermediate in trehalose metabolism, has been shown to regulate embryonic and vegetative development, flowering time, meristem determinacy, and cell fate specification in plants. T6P acts as a global regulator of metabolism and transcription promoting plant growth and triggering developmental phase transitions in response to sugar availability. Among the T6P targets are members of the Sucrose-non-fermenting1-related kinase1 (SnRK1) family, which are sensors of energy availability and inhibit plant growth and development during metabolic stress to maintain energy homeostasis. In this review, we will discuss the opposite roles of the sugar metabolite T6P and the SnRK1 kinases in the regulation of developmental phase transitions in response to carbohydrate levels. We will focus on how these two global regulators of metabolic processes integrate environmental cues and interact with hormonal signaling pathways to modulate plant development. PMID:24744765

  18. Absolute Radiometric Calibration of the GÖKTÜRK-2 Satellite Sensor Using Tuz GÖLÜ (landnet Site) from Ndvi Perspective

    NASA Astrophysics Data System (ADS)

    Sakarya, Ufuk; Hakkı Demirhan, İsmail; Seda Deveci, Hüsne; Teke, Mustafa; Demirkesen, Can; Küpçü, Ramazan; Feray Öztoprak, A.; Efendioğlu, Mehmet; Fehmi Şimşek, F.; Berke, Erdinç; Zübeyde Gürbüz, Sevgi

    2016-06-01

    TÜBİTAK UZAY has conducted a research study on the use of space-based satellite resources for several aspects of agriculture. Especially, there are two precision agriculture related projects: HASSAS (Widespread application of sustainable precision agriculture practices in Southeastern Anatolia Project Region (GAP) Project) and AKTAR (Smart Agriculture Feasibility Project). The HASSAS project aims to study development of precision agriculture practice in GAP region. Multi-spectral satellite imagery and aerial hyperspectral data along with ground measurements was collected to analyze data in an information system. AKTAR aims to develop models for irrigation, fertilization and spectral signatures of crops in Inner Anatolia. By the end of the project precision agriculture practices to control irrigation, fertilization, pesticide and estimation of crop yield will be developed. Analyzing the phenology of crops using NDVI is critical for the projects. For this reason, absolute radiometric calibration of the Red and NIR bands in space-based satellite sensors is an important issue. The Göktürk-2 satellite is an earth observation satellite which was designed and built in Turkey and was launched in 2012. The Göktürk-2 satellite sensor has a resolution 2.5 meters in panchromatic and 5 meters in R/G/B/NIR bands. The absolute radiometric calibration of the Göktürk-2 satellite sensor was performed via the ground-based measurements - spectra-radiometer, sun photometer, and meteorological station- in Tuz Gölü cal/val site in 2015. In this paper, the first ground-based absolute radiometric calibration results of the Göktürk-2 satellite sensor using Tuz Gölü is demonstrated. The absolute radiometric calibration results of this paper are compared with the published cross-calibration results of the Göktürk-2 satellite sensor utilizing Landsat 8 imagery. According to the experimental comparison results, the Göktürk-2 satellite sensor coefficients for red and NIR bands

  19. Peptidoglycan metabolism is controlled by the WalRK (YycFG) and PhoPR two-component systems in phosphate-limited Bacillus subtilis cells.

    PubMed

    Bisicchia, Paola; Lioliou, Efthimia; Noone, David; Salzberg, Letal I; Botella, Eric; Hübner, Sebastian; Devine, Kevin M

    2010-02-01

    In Bacillus subtilis, the WalRK (YycFG) two-component system controls peptidoglycan metabolism in exponentially growing cells while PhoPR controls the response to phosphate limitation. Here we examine the roles of WalRK and PhoPR in peptidoglycan metabolism in phosphate-limited cells. We show that B. subtilis cells remain viable in a phosphate-limited state for an extended period and resume growth rapidly upon phosphate addition, even in the absence of a PhoPR-mediated response. Peptidoglycan synthesis occurs in phosphate-limited wild-type cells at approximately 27% the rate of exponentially growing cells, and at approximately 18% the rate of exponentially growing cells in the absence of PhoPR. In phosphate-limited cells, the WalRK regulon genes yocH, cwlO(yvcE), lytE and ydjM are expressed in a manner that is dependent on the WalR recognition sequence and deleting these genes individually reduces the rate of peptidoglycan synthesis. We show that ydjM expression can be activated by PhoP approximately P in vitro and that PhoP occupies its promoter in phosphate-limited cells. However, iseA(yoeB) expression cannot be repressed by PhoP approximately P in vitro, but can be repressed by non-phosphorylated WalR in vitro. Therefore, we conclude that peptidoglycan metabolism is controlled by both WalRK and PhoPR in phosphate-limited B. subtilis cells. PMID:20487291

  20. The SnRK2-APC/CTE regulatory module mediates the antagonistic action of gibberellic acid and abscisic acid pathways

    PubMed Central

    Lin, Qibing; Wu, Fuqing; Sheng, Peike; Zhang, Zhe; Zhang, Xin; Guo, Xiuping; Wang, Jiulin; Cheng, Zhijun; Wang, Jie; Wang, Haiyang; Wan, Jianmin

    2015-01-01

    Abscisic acid (ABA) and gibberellic acid (GA) antagonistically regulate many developmental processes and responses to biotic or abiotic stresses in higher plants. However, the molecular mechanism underlying this antagonism is still poorly understood. Here, we show that loss-of-function mutation in rice Tiller Enhancer (TE), an activator of the APC/CTE complex, causes hypersensitivity and hyposensitivity to ABA and GA, respectively. We find that TE physically interacts with ABA receptor OsPYL/RCARs and promotes their degradation by the proteasome. Genetic analysis also shows OsPYL/RCARs act downstream of TE in mediating ABA responses. Conversely, ABA inhibits APC/CTE activity by phosphorylating TE through activating the SNF1-related protein kinases (SnRK2s), which may interrupt the interaction between TE and OsPYL/RCARs and subsequently stabilize OsPYL/RCARs. In contrast, GA can reduce the level of SnRK2s and may promote APC/CTE-mediated degradation of OsPYL/RCARs. Thus, we propose that the SnRK2-APC/CTE regulatory module represents a regulatory hub underlying the antagonistic action of GA and ABA in plants. PMID:26272249

  1. Arabidopsis RZFP34/CHYR1, a Ubiquitin E3 Ligase, Regulates Stomatal Movement and Drought Tolerance via SnRK2.6-Mediated Phosphorylation[OPEN

    PubMed Central

    2015-01-01

    Abscisic acid (ABA) is a phytohormone that plays a fundamental role in plant development and stress response, especially in the regulation of stomatal closure in response to water deficit stress. The signal transduction that occurs in response to ABA and drought stress is mediated by protein phosphorylation and ubiquitination. This research identified Arabidopsis thaliana RING ZINC-FINGER PROTEIN34 (RZP34; renamed here as CHY ZINC-FINGER AND RING PROTEIN1 [CHYR1]) as an ubiquitin E3 ligase. CHYR1 expression was significantly induced by ABA and drought, and along with its corresponding protein, was expressed mainly in vascular tissues and stomata. Analysis of CHYR1 gain-of-function and loss-of-function plants revealed that CHYR1 promotes ABA-induced stomatal closure, reactive oxygen species production, and plant drought tolerance. Furthermore, CHYR1 interacted with SNF1-RELATED PROTEIN KINASE2 (SnRK2) kinases and could be phosphorylated by SnRK2.6 on the Thr-178 residue. Overexpression of CHYR1T178A, a phosphorylation-deficient mutant, interfered with the proper function of CHYR1, whereas CHYR1T178D phenocopied the gain of function of CHYR1. Thus, this study identified a RING-type ubiquitin E3 ligase that functions positively in ABA and drought responses and detailed how its ubiquitin E3 ligase activity is regulated by SnRK2.6-mediated protein phosphorylation. PMID:26508764

  2. Cloning of Gossypium hirsutum Sucrose Non-Fermenting 1-Related Protein Kinase 2 Gene (GhSnRK2) and Its Overexpression in Transgenic Arabidopsis Escalates Drought and Low Temperature Tolerance

    PubMed Central

    Bello, Babatunde; Zhang, Xueyan; Liu, Chuanliang; Yang, Zhaoen; Yang, Zuoren; Wang, Qianhua; Zhao, Ge; Li, Fuguang

    2014-01-01

    The molecular mechanisms of stress tolerance and the use of modern genetics approaches for the improvement of drought stress tolerance have been major focuses of plant molecular biologists. In the present study, we cloned the Gossypium hirsutum sucrose non-fermenting 1-related protein kinase 2 (GhSnRK2) gene and investigated its functions in transgenic Arabidopsis. We further elucidated the function of this gene in transgenic cotton using virus-induced gene silencing (VIGS) techniques. We hypothesized that GhSnRK2 participates in the stress signaling pathway and elucidated its role in enhancing stress tolerance in plants via various stress-related pathways and stress-responsive genes. We determined that the subcellular localization of the GhSnRK2-green fluorescent protein (GFP) was localized in the nuclei and cytoplasm. In contrast to wild-type plants, transgenic plants overexpressing GhSnRK2 exhibited increased tolerance to drought, cold, abscisic acid and salt stresses, suggesting that GhSnRK2 acts as a positive regulator in response to cold and drought stresses. Plants overexpressing GhSnRK2 displayed evidence of reduced water loss, turgor regulation, elevated relative water content, biomass, and proline accumulation. qRT-PCR analysis of GhSnRK2 expression suggested that this gene may function in diverse tissues. Under normal and stress conditions, the expression levels of stress-inducible genes, such as AtRD29A, AtRD29B, AtP5CS1, AtABI3, AtCBF1, and AtABI5, were increased in the GhSnRK2-overexpressing plants compared to the wild-type plants. GhSnRK2 gene silencing alleviated drought tolerance in cotton plants, indicating that VIGS technique can certainly be used as an effective means to examine gene function by knocking down the expression of distinctly expressed genes. The results of this study suggested that the GhSnRK2 gene, when incorporated into Arabidopsis, functions in positive responses to drought stress and in low temperature tolerance. PMID:25393623

  3. Transcriptional regulation of SlPYL, SlPP2C, and SlSnRK2 gene families encoding ABA signal core components during tomato fruit development and drought stress

    PubMed Central

    Sun, Liang; Wang, Yan-Ping; Chen, Pei; Ren, Jie; Ji, Kai; Li, Qian; Li, Ping; Dai, Sheng-Jie; Leng, Ping

    2011-01-01

    In order to characterize the potential transcriptional regulation of core components of abscisic acid (ABA) signal transduction in tomato fruit development and drought stress, eight SlPYL (ABA receptor), seven SlPP2C (type 2C protein phosphatase), and eight SlSnRK2 (subfamily 2 of SNF1-related kinases) full-length cDNA sequences were isolated from the tomato nucleotide database of NCBI GenBank. All SlPYL, SlPP2C, and SlSnRK2 genes obtained are homologous to Arabidopsis AtPYL, AtPP2C, and AtSnRK2 genes, respectively. Based on phylogenetic analysis, SlPYLs and SlSnRK2s were clustered into three subfamilies/subclasses, and all SlPP2Cs belonged to PP2C group A. Within the SlPYL gene family, SlPYL1, SlPYL2, SlPYL3, and SlPYL6 were the major genes involved in the regulation of fruit development. Among them, SlPYL1 and SlPYL2 were expressed at high levels throughout the process of fruit development and ripening; SlPYL3 was strongly expressed at the immature green (IM) and mature green (MG) stages, while SlPYL6 was expressed strongly at the IM and red ripe (RR) stages. Within the SlPP2C gene family, the expression of SlPP2C, SlPP2C3, and SlPP2C4 increased after the MG stage; SlPP2C1 and SlPP2C5 peaked at the B3 stage, while SlPP2C2 and SlPP2C6 changed little during fruit development. Within the SlSnRK2 gene family, the expression of SlSnRK2.2, SlSnRK2.3, SlSnRK2.4, and SlSnRK2C was higher than that of other members during fruit development. Additionally, most SlPYL genes were down-regulated, while most SlPP2C and SlSnRK2 genes were up-regulated by dehydration in tomato leaf. PMID:21873532

  4. Source/sink interactions underpin crop yield: the case for trehalose 6-phosphate/SnRK1 in improvement of wheat.

    PubMed

    Lawlor, David W; Paul, Matthew J

    2014-01-01

    Considerable interest has been evoked by the analysis of the regulatory pathway in carbohydrate metabolism and cell growth involving the non-reducing disaccharide trehalose (TRE). TRE is at small concentrations in mesophytes such as Arabidopsis thaliana and Triticum aestivum, excluding a role in osmoregulation once suggested for it. Studies of TRE metabolism, and genetic modification of it, have shown a very wide and more important role of the pathway in regulation of many processes in development, growth, and photosynthesis. It has now been established that rather than TRE, it is trehalose 6-phosphate (T6P) which has such profound effects. T6P is the intermediary in TRE synthesis formed from glucose-6-phosphate and UDP-glucose, derived from sucrose, by the action of trehalose phosphate synthase. The concentration of T6P is determined both by the rate of synthesis, which depends on the sucrose concentration, and also by the rate of breakdown by trehalose-6-phosphate phosphatase which produces TRE. Changing T6P concentrations by genetically modifying the enzymes of synthesis and breakdown has altered photosynthesis, sugar metabolism, growth, and development which affect responses to, and recovery from, environmental factors. Many of the effects of T6P on metabolism and growth occur via the interaction of T6P with the SnRK1 protein kinase system. T6P inhibits the activity of SnRK1, which de-represses genes encoding proteins involved in anabolism. Consequently, a large concentration of sucrose increases T6P and thereby inhibits SnRK1, so stimulating growth of cells and their metabolic activity. The T6P/SnRK1 mechanism offers an important new view of how the distribution of assimilates to organs, such as developing grains in cereal plants, is achieved. This review briefly summarizes the factors determining, and limiting, yield of wheat (particularly mass/grain which is highly conserved) and considers how T6P/SnRK1 might function to determine grain yield and might be

  5. Identification of variables and value optimization for optimum lipase production by Bacillus pumilus RK31 using statistical methodology.

    PubMed

    Kumar, Rakesh; Mahajan, Shivani; Kumar, Arun; Singh, Deepak

    2011-01-31

    In an effort to optimize the medium components, the statistical methodology was applied to achieve the optimum lipase production under shake flask conditions. The study was conducted in three steps on newly isolated Bacillus pumilus RK 31. In the first step, 12 different variables viz., Glucose, Olive oil, Yeast extract, Peptone, Tween 80, KH(2)PO(4), MgSO(4), NaNO(3), CaCl(2), Temperature, pH and Inoculum size were used to identify the most significant variables affecting lipase production using Plackett-Burman statistical design. Variance analysis showed that Olive oil, Tween 80 and KH(2)PO(4) played significant role in lipase production. In the second step, the values of the above-identified three variables were optimized by central composite design using three-level-three-factor approach. The optimum values of Olive oil, Tween 80 and KH(2)PO(4) were found to be 10.0ml/l, 5.0ml/l and 8.0g/l, respectively. KH(2)PO(4) was found to be responsible for maximum lipase production of 5.59IU/ml, experimental and 5.03IU/ml, predicted. In the third step, the optimum predicted values of the three factors and lipase production were verified by experimental approach. The amount of lipase produced in the designated medium was in agreement with that of predicted values by statistical method. PMID:20601261

  6. Impact of the ATATÜRK DAM Lake on Agro-Meteorological Aspects of the Southeastern Anatolia Region Using Remote Sensing and GIS Analysis

    NASA Astrophysics Data System (ADS)

    Ozcan, O.; Bookhagen, B.; Musaoglu, N.

    2012-07-01

    The Atatürk Dam is the fourth largest clay-cored rock fill dam in the world. It was constructed on the Euphrates River located in semi-arid Southeastern Turkey in the 1980s as the central component of a large-scale regional development project for the Southeastern Anatolia region (referred to as GAP). The construction began in 1983 and was completed in 1990. The dam and the hydroelectric power plant, which went into service after filling up the reservoir was accomplished in 1992. The Atatürk Dam, which has a height of 169 m, a total storage capacity of 48.7 million m3, and a surface area of about 817 km2 plays an important role in the development of Turkey's energy and agriculture sectors. In this study, the spatial and temporal impacts of the Atatürk Dam on agro-meteorological aspects of the Southeastern Anatolia region have been investigated. Change detection and environmental impacts due to water-reserve changes in Atatürk Dam Lake have been determined and evaluated using multi-temporal Landsat satellite imageries and meteorological datasets within a period of 1984 to 2011. These time series have been evaluated for three time periods. Dam construction period constitutes the first part of the study. Land cover/use changes especially on agricultural fields under the Atatürk Dam Lake and its vicinity have been identified between the periods of 1984 to 1992. The second period comprises the 10-year period after the completion of filling up the reservoir in 1992. At this period, Landsat and meteorological time-series analyses are examined to assess the impact of the Atatürk Dam Lake on selected irrigated agricultural areas. For the last 9-year period from 2002 to 2011, the relationships between seasonal water-reserve changes and irrigated plains under changing climatic factors primarily driving vegetation activity (monthly, seasonal, and annual fluctuations of rainfall rate, air temperature, humidity) on the watershed have been investigated using a 30-year

  7. Diagnostic tests for kala-azar: a multi-centre study of the freeze-dried DAT, rK39 strip test and KAtex in East Africa and the Indian subcontinent.

    PubMed

    Boelaert, M; El-Safi, S; Hailu, A; Mukhtar, M; Rijal, S; Sundar, S; Wasunna, M; Aseffa, A; Mbui, J; Menten, J; Desjeux, P; Peeling, R W

    2008-01-01

    Three diagnostic tests for visceral leishmaniasis (VL), the freeze-dried direct agglutination test (FD-DAT), the rK39 dipstick and a urine latex antigen test (KAtex), were evaluated for use in primary care in East Africa and the Indian subcontinent. Clinical suspects were prospectively recruited and tissue, blood and urine samples were taken. Direct microscopic examination of tissue smear, and FD-DAT, rK39 and KAtex were performed. Sensitivity and specificity with 95% credible intervals were estimated using Bayesian latent class analysis. On the Indian subcontinent both the FD-DAT and the rK39 strip test exceeded the 95% sensitivity and 90% specificity target, but not so in East Africa. Sensitivity of the FD-DAT was high in Ethiopia and Kenya but lower in Sudan, while its specificity was below 90% in Kenya. Sensitivity of the rK39 was below 80% in the three countries, and its specificity was only 70% in Ethiopia. KAtex showed moderate to very low sensitivity in all countries. FD-DAT and rK39 can be recommended for clinical practice on the Indian subcontinent. In East Africa, their clinical use should be carefully monitored. More work is needed to improve existing formats, and to develop better VL diagnostics. PMID:17942129

  8. Detecting broken struts of a Björk-Shiley heart valve using ultrasound: a feasibility study.

    PubMed

    van Neer, P L M J; Bouakaz, A; Vlaanderen, E; de Hart, J; van de Vosse, F N; van der Steen, A F W; de Jong, N

    2006-04-01

    The Björk-Shiley (BScc) mechanical heart valve has extensively been used in surgery from 1979 to 1986. There is, compared with equivalent valve types, increased occurrence of unexpected mechanical failure of the outlet strut of the valve, with a high incidence of mortality, when it occurs. Many approaches have been attempted to noninvasively determine BScc valve integrity. None of the approaches resulted in adequate assessment, mostly due to a lack of either sensitivity or specificity demonstrated in in vitro and/or in vivo studies. In our study we analyze leg movement of the BScc valves outlet strut during the cardiac cycle with ultrasound. For a broken strut, the movement of both legs will be significantly different, whereas the difference will be negligible for an intact strut. BScc valves were mounted in the mitral position in an in vitro pulse duplicator system. A focused single-element transducer was used to direct ultrasound on a leg of the outlet strut. Correlation-based time delay estimation was used to estimate differences in time of flight of the outlet strut echoes to determine outlet strut leg movement. The movement of an intact valve and a valve with a single-leg fracture with both ends grating against each other (SLF), the most difficult fracture to diagnose, has been studied. The results showed no significant difference in movement between both legs of the outlet strut of the intact BScc valve (amplitude of movement 9.2 microm +/- 0.1 microm). Whereas for the defective valve, the amplitude of movement of the broken leg of the SLF valve was 12 microm +/- 1.6 microm vs. 8.6 microm +/- 0.1 microm for the intact leg. In conclusion, the proposed method has shown to be feasible in vitro and has potentials for in vivo detection of BScc valve outlet strut fracture. PMID:16616597

  9. Discrepancies in the determination of sperm concentration using Bürker-Türk, Thoma and Makler counting chambers.

    PubMed

    Christensen, P; Stryhn, H; Hansen, C

    2005-03-01

    Determination of sperm concentration by use of a haemocytometer or counting chamber is an important step in semen evaluation and is also used for calibration or validation of instruments. Three experiments were carried out to determine the precision and accuracy of the Makler chamber, the Thoma haemocytometers (50 and 100 microm deep, TH-50, TH-100) and the Bürker-Türk (BT) haemocytometer. The first experiment confirmed that precision in sperm count by use of the haemocytometers (TH-50 and BT) can be increased if a higher number of sperm are counted. In contrast, the precision of the Makler chamber was relatively unaffected by the number of sperm counted and the coefficient of variation for this chamber was significantly (P < 0.05) higher than for the two haemocytometers. Experiment 2 confirmed the low precision of the Makler chamber and also showed that the TH-50 haemocytometer underestimated sperm concentration by approximately 25% in comparison to the Makler chamber and the BT haemocytometer. Experiment 3 demonstrated a slight underestimation of sperm count by the TH-100 haemocytometer in comparison to the BT haemocytometer, but both haemocytometers yielded acceptable precision (coefficients of variation were 10.4% and 9.4%, respectively). In comparison, the precision of the Makler chamber was significantly poorer (coefficient of variation 18.6%). When used for validation of a flow cytometric method which determines sperm concentration, the Makler chamber caused a higher degree of scattering of the points around the regression line than when the flow cytometric method was validated against the BT haemocytometer. It thus appears that the poor precision of the Makler chamber also affects the accuracy. It is concluded that duplicate counts by at least two technicians is recommended to achieve high precision but, that particular caution should be exerted with regard to the precision and accuracy of the used counting device. PMID:15710187

  10. Earthquake imprints on a lacustrine deltaic system: the Kürk Delta along the East Anatolian Fault (Turkey)

    NASA Astrophysics Data System (ADS)

    Hubert-Ferrari, Aurélia; El-Ouahabi, Meriam; Garcia-Moreno, David; Avsar, Ulas; Altinok, Sevgi; Schmidt, Sabine; Cagatay, Namik

    2016-04-01

    Delta contains a sedimentary record primarily indicative of water level changes, but particularly sensitive to earthquake shaking, which results generally in soft-sediment-deformation structures. The Kürk Delta adjacent to a major strike-slip fault displays this type of deformation (Hempton and Dewey, 1983) as well as other types of earthquake fingerprints that are specifically investigated. This lacustrine delta stands at the south-western extremity of the Hazar Lake and is bound by the East Anatolian Fault (EAF), which generated earthquakes of magnitude 7 in eastern Turkey. Water level changes and earthquake shaking affecting the Kurk Delta have been reevaluated combining geophysical data (seismic-reflection profiles and side-scan sonar), remote sensing images, historical data, onland outcrops and offshore coring. The history of water level changes provides a temporal framework regarding the sedimentological record. In addition to the commonly soft-sediment-deformation previously documented, the onland outcrops reveal a record of deformation (faults and clastic dykes) linked to large earthquake-induced liquefactions. The recurrent liquefaction structures can be used to obtain a paleoseismological record. Five event horizons were identified that could be linked to historical earthquakes occurring in the last 1000 years along the EAF. Sedimentary cores sampling the most recent subaqueous sedimentation revealed the occurrence of another type of earthquake fingerprint. Based on radionuclide dating (137Cs and 210Pb), two major sedimentary events were attributed to the 1874-1875 earthquake sequence along the EAF. Their sedimentological characteristics were inferred based X-ray imagery, XRD, LOI, grain-size distribution, geophysical measurements. The events are interpreted to be hyperpycnal deposits linked to post-seismic sediment reworking of earthquake-triggered landslides. A time constraint regarding this sediment remobilization process could be achieved thanks to

  11. A novel ARID DNA-binding protein interacts with SymRK and is expressed during early nodule development in Lotus japonicus.

    PubMed

    Zhu, Hui; Chen, Tao; Zhu, Maosheng; Fang, Qing; Kang, Heng; Hong, Zonglie; Zhang, Zhongming

    2008-09-01

    During the establishment of symbiosis in legume roots, the rhizobial Nod factor signal is perceived by the host cells via receptor-like kinases, including SymRK. The NODULE INCEPTION (NIN) gene in Lotus japonicus is required for rhizobial entry into root cells and for nodule organogenesis. We describe here a novel DNA-binding protein from L. japonicus, referred to as SIP1, because it was identified as a SymRK-interacting protein. SIP1 contains a conserved AT-rich interaction domain (ARID) and represents a unique member of the ARID-containing proteins in plants. The C terminus of SIP1 was found to be responsible for its interaction with the kinase domain of SymRK and for homodimerization in the absence of DNA. SIP1 specifically binds to the promoter of LjNIN but not to that of LjCBP1 (a calcium-binding protein gene), both of which are known to be inducible by Nod factors. SIP1 recognizes two of the three AT-rich domains present in the NIN gene promoter. Deletion of one of the AT-rich domains at the NIN promoter diminishes the binding of SIP1 to the NIN promoter. The protein is localized to the nuclei when expressed as a red fluorescence fusion protein in the onion (Allium cepa) epidermal cells. The SIP1 gene is expressed constitutively in the uninfected roots, and its expression levels are elevated after infection by Mesorhizobium loti. It is proposed that SIP1 may be required for the expression of NIN and involved in the initial communications between the rhizobia and the host root cells. PMID:18633121

  12. SnRK1 Isoforms AKIN10 and AKIN11 Are Differentially Regulated in Arabidopsis Plants under Phosphate Starvation1[C][OA

    PubMed Central

    Fragoso, Selene; Espíndola, Laura; Páez-Valencia, Julio; Gamboa, Alicia; Camacho, Yolanda; Martínez-Barajas, Eleazar; Coello, Patricia

    2009-01-01

    During phosphate starvation, Snf1-related kinase 1 (SnRK1) activity significantly decreases compared with plants growing under normal nutritional conditions. An analysis of the expression of the genes encoding for the catalytic subunits of SnRK1 showed that these subunits were not affected by phosphate starvation. Transgenic Arabidopsis (Arabidopsis thaliana) plants overexpressing the AKIN10 and AKIN11 catalytic subunits fused with green fluorescent protein (GFP) were produced, and their localizations were mainly chloroplastic with low but detectable signals in the cytoplasm. These data were corroborated with an immunocytochemistry analysis using leaf and root sections with an anti-AKIN10/AKIN11 antibody. The SnRK1 activity in transgenic plants overexpressing AKIN11-GFP was reduced by 35% to 40% in phosphate starvation, in contrast with the results observed in plants overexpressing AKIN10-GFP, which increased the activity by 100%. No differences in activity were observed in plants growing in phosphate-sufficient conditions. Biochemical analysis of the proteins indicated that AKIN11 is specifically degraded under these limited conditions and that the increase in AKIN10-GFP activity was not due to the phosphorylation of threonine-175. These results are consistent with an important role of AKIN10 in signaling during phosphate starvation. Moreover, akin10 mutant plants were deficient in starch mobilization at night during inorganic phosphate starvation, and under this condition several genes were up-regulated and down-regulated, indicating their important roles in the control of general transcription. This finding reveals novel roles for the different catalytic subunits during phosphate starvation. PMID:19211700

  13. Arabidopsis thaliana Remorins Interact with SnRK1 and Play a Role in Susceptibility to Beet Curly Top Virus and Beet Severe Curly Top Virus.

    PubMed

    Son, Seungmin; Oh, Chang Jae; An, Chung Sun

    2014-09-01

    Remorins, a family of plant-specific proteins containing a variable N-terminal region and conserved C-terminal domain, play a role in various biotic and abiotic stresses, including host-microbe interactions. However, their functions remain to be completely elucidated, especially for the Arabidopsis thaliana remorin group 4 (AtREM4). To elucidate the role of remorins in Arabidopsis, we first showed that AtREM4s have typical molecular characteristics of the remorins, such as induction by various types of biotic and abiotic stresses, localization in plasma membrane and homo- and hetero-oligomeric interaction. Next, we showed that their loss-of-function mutants displayed reduced susceptibility to geminiviruses, Beet Curly Top Virus and Beet Severe Curly Top Virus, while overexpressors enhanced susceptibility. Moreover, we found that they interacted with SnRK1, which phosphorylated AtREM4.1, and were degraded by the 26S proteasome pathway. These results suggest that AtREM4s may be involved in the SnRK1-mediated signaling pathway and play a role as positive regulators of the cell cycle during geminivirus infection. PMID:25289013

  14. Arabidopsis thaliana Remorins Interact with SnRK1 and Play a Role in Susceptibility to Beet Curly Top Virus and Beet Severe Curly Top Virus

    PubMed Central

    Son, Seungmin; Oh, Chang Jae; An, Chung Sun

    2014-01-01

    Remorins, a family of plant-specific proteins containing a variable N-terminal region and conserved C-terminal domain, play a role in various biotic and abiotic stresses, including host-microbe interactions. However, their functions remain to be completely elucidated, especially for the Arabidopsis thaliana remorin group 4 (AtREM4). To elucidate the role of remorins in Arabidopsis, we first showed that AtREM4s have typical molecular characteristics of the remorins, such as induction by various types of biotic and abiotic stresses, localization in plasma membrane and homo- and hetero-oligomeric interaction. Next, we showed that their loss-of-function mutants displayed reduced susceptibility to geminiviruses, Beet Curly Top Virus and Beet Severe Curly Top Virus, while overexpressors enhanced susceptibility. Moreover, we found that they interacted with SnRK1, which phosphorylated AtREM4.1, and were degraded by the 26S proteasome pathway. These results suggest that AtREM4s may be involved in the SnRK1-mediated signaling pathway and play a role as positive regulators of the cell cycle during geminivirus infection. PMID:25289013

  15. Purification and cDNA cloning of SAPKK3, the major activator of RK/p38 in stress- and cytokine-stimulated monocytes and epithelial cells.

    PubMed Central

    Cuenda, A; Alonso, G; Morrice, N; Jones, M; Meier, R; Cohen, P; Nebreda, A R

    1996-01-01

    Two chromatographically distinct stress-activated protein kinase kinases (SAPKKs) have been identified in several mammalian cells, termed SAPKK2 and SAPKK3, which activate the MAP kinase family member RK/p38 but not JNK/SAPK in vitro. Here we demonstrate that SAPKK2 is identical or very closely related to the MAP kinase kinase family member MKK3. However, under our assay conditions, SAPKK3 was the major activator of RK/p38 detected in extracts prepared from stress- or interleukin-1-stimulated epithelial (KB) cells, from bacterial lipopolysaccharide and tumour necrosis factor alpha-stimulated THP1 monocytes or from rabbit skeletal muscle. The activated form of SAPKK3 was purified from muscle to near homogeneity, and tryptic peptide sequences were used to clone human and murine cDNAs encoding this enzyme. Human SAPKK3 comprised 334 amino acids and was 78% identical to MKK3. The murine and human SAPKK3 were 97% identical in their amino acid sequences. We also cloned a different murine cDNA that appears to encode a SAPKK3 protein truncated at the N-terminus. SAPKK3 is identical to the recently cloned MKK6. Images PMID:8861944

  16. Differential regulation of the MAP, SAP and RK/p38 kinases by Pyst1, a novel cytosolic dual-specificity phosphatase.

    PubMed Central

    Groom, L A; Sneddon, A A; Alessi, D R; Dowd, S; Keyse, S M

    1996-01-01

    The Pyst1 and Pyst2 mRNAs encode closely related proteins, which are novel members of a family of dual-specificity MAP kinase phosphatases typified by CL100/MKP-1. Pyst1 is expressed constitutively in human skin fibroblasts and, in contrast to other members of this family of enzymes, its mRNA is not inducible by either stress or mitogens. Furthermore, unlike the nuclear CL100 protein, Pyst1 is localized in the cytoplasm of transfected Cos-1 cells. Like CL100/ MKP-1, Pyst1 dephosphorylates and inactivates MAP kinase in vitro and in vivo. In addition, Pyst1 is able to form a physical complex with endogenous MAP kinase in Cos-1 cells. However, unlike CL100, Pyst1 displays very low activity towards the stress-activated protein kinases (SAPKs) or RK/p38 in vitro, indicating that these kinases are not physiological substrates for Pyst1. This specificity is underlined by the inability of Pyst1 to block either the stress-mediated activation of the JNK-1 SAP kinase or RK/p38 in vivo, or to inhibit nuclear signalling events mediated by the SAP kinases in response to UV radiation. Our results provide the first evidence that the members of the MAP kinase family of enzymes are differentially regulated by dual-specificity phosphatases and also indicate that the MAP kinases may be regulated by different members of this family of enzymes depending on their subcellular location. Images PMID:8670865

  17. Cloning and expression analyses of Sucrose non-fermenting-1-Related Kinase 1 (SnRK1b) gene during development of sorghum and maize endosperm, and its implicated role in sugar-to-starch metabolic transition

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A full-length cDNA clone, SbSnRK1b (1530 bp, GenBank accession no. EF544393), encoding a putative serine/threonine protein kinase homologue of yeast (Saccharomyces cerevisiae) SNF1, was isolated from developing endosperm of sorghum [Sorghum bicolor (Moench) L.]. Multiple sequence alignment data show...

  18. The Eukaryotic-Like Ser/Thr Kinase PrkC Regulates the Essential WalRK Two-Component System in Bacillus subtilis

    PubMed Central

    Dworkin, Jonathan

    2015-01-01

    Most bacteria contain both eukaryotic-like Ser/Thr kinases (eSTKs) and eukaryotic-like Ser/Thr phosphatases (eSTPs). Their role in bacterial physiology is not currently well understood in large part because the conditions where the eSTKs are active are generally not known. However, all sequenced Gram-positive bacteria have a highly conserved eSTK with extracellular PASTA repeats that bind cell wall derived muropeptides. Here, we report that in the Gram-positive bacterium Bacillus subtilis, the PASTA-containing eSTK PrkC and its cognate eSTP PrpC converge with the essential WalRK two-component system to regulate WalR regulon genes involved in cell wall metabolism. By continuously monitoring gene expression throughout growth, we consistently find a large PrkC-dependent effect on expression of several different WalR regulon genes in early stationary phase, including both those that are activated by WalR (yocH) as well as those that are repressed (iseA, pdaC). We demonstrate that PrkC phosphorylates WalR in vitro and in vivo on a single Thr residue located in the receiver domain. Although the phosphorylated region of the receiver domain is highly conserved among several B. subtilis response regulators, PrkC displays specificity for WalR in vitro. Consistently, strains expressing a nonphosphorylatable WalR point mutant strongly reduce both PrkC dependent activation and repression of yocH, iseA, and pdaC. This suggests a model where the eSTK PrkC regulates the essential WalRK two-component signaling system by direct phosphorylation of WalR Thr101, resulting in the regulation of WalR regulon genes involved in cell wall metabolism in stationary phase. As both the eSTK PrkC and the essential WalRK two-component system are highly conserved in Gram-positive bacteria, these results may be applicable to further understanding the role of eSTKs in Gram-positive physiology and cell wall metabolism. PMID:26102633

  19. The Eukaryotic-Like Ser/Thr Kinase PrkC Regulates the Essential WalRK Two-Component System in Bacillus subtilis.

    PubMed

    Libby, Elizabeth A; Goss, Lindsie A; Dworkin, Jonathan

    2015-06-01

    Most bacteria contain both eukaryotic-like Ser/Thr kinases (eSTKs) and eukaryotic-like Ser/Thr phosphatases (eSTPs). Their role in bacterial physiology is not currently well understood in large part because the conditions where the eSTKs are active are generally not known. However, all sequenced Gram-positive bacteria have a highly conserved eSTK with extracellular PASTA repeats that bind cell wall derived muropeptides. Here, we report that in the Gram-positive bacterium Bacillus subtilis, the PASTA-containing eSTK PrkC and its cognate eSTP PrpC converge with the essential WalRK two-component system to regulate WalR regulon genes involved in cell wall metabolism. By continuously monitoring gene expression throughout growth, we consistently find a large PrkC-dependent effect on expression of several different WalR regulon genes in early stationary phase, including both those that are activated by WalR (yocH) as well as those that are repressed (iseA, pdaC). We demonstrate that PrkC phosphorylates WalR in vitro and in vivo on a single Thr residue located in the receiver domain. Although the phosphorylated region of the receiver domain is highly conserved among several B. subtilis response regulators, PrkC displays specificity for WalR in vitro. Consistently, strains expressing a nonphosphorylatable WalR point mutant strongly reduce both PrkC dependent activation and repression of yocH, iseA, and pdaC. This suggests a model where the eSTK PrkC regulates the essential WalRK two-component signaling system by direct phosphorylation of WalR Thr101, resulting in the regulation of WalR regulon genes involved in cell wall metabolism in stationary phase. As both the eSTK PrkC and the essential WalRK two-component system are highly conserved in Gram-positive bacteria, these results may be applicable to further understanding the role of eSTKs in Gram-positive physiology and cell wall metabolism. PMID:26102633

  20. Growth Arrest by Trehalose-6-Phosphate: An Astonishing Case of Primary Metabolite Control over Growth by Way of the SnRK1 Signaling Pathway1[C][W][OA

    PubMed Central

    Delatte, Thierry L.; Sedijani, Prapti; Kondou, Youichi; Matsui, Minami; de Jong, Gerhardus J.; Somsen, Govert W.; Wiese-Klinkenberg, Anika; Primavesi, Lucia F.; Paul, Matthew J.; Schluepmann, Henriette

    2011-01-01

    The strong regulation of plant carbon allocation and growth by trehalose metabolism is important for our understanding of the mechanisms that determine growth and yield, with obvious applications in crop improvement. To gain further insight on the growth arrest by trehalose feeding, we first established that starch-deficient seedlings of the plastidic phosphoglucomutase1 mutant were similarly affected as the wild type on trehalose. Starch accumulation in the source cotyledons, therefore, did not cause starvation and consequent growth arrest in the growing zones. We then screened the FOX collection of Arabidopsis (Arabidopsis thaliana) expressing full-length cDNAs for seedling resistance to 100 mm trehalose. Three independent transgenic lines were identified with dominant segregation of the trehalose resistance trait that overexpress the bZIP11 (for basic region/leucine zipper motif) transcription factor. The resistance of these lines to trehalose could not be explained simply through enhanced trehalase activity or through inhibition of bZIP11 translation. Instead, trehalose-6-phosphate (T6P) accumulation was much increased in bZIP11-overexpressing lines, suggesting that these lines may be insensitive to the effects of T6P. T6P is known to inhibit the central stress-integrating kinase SnRK1 (KIN10) activity. We confirmed that this holds true in extracts from seedlings grown on trehalose, then showed that two independent transgenic lines overexpressing KIN10 were insensitive to trehalose. Moreover, the expression of marker genes known to be jointly controlled by SnRK1 activity and bZIP11 was consistent with low SnRK1 or bZIP11 activity in seedlings on trehalose. These results reveal an astonishing case of primary metabolite control over growth by way of the SnRK1 signaling pathway involving T6P, SnRK1, and bZIP11. PMID:21753116

  1. Numerical analysis of the supercontinuum spectrum generation in a couple of photonic crystal fibers with different structure by using the RK4IP method

    NASA Astrophysics Data System (ADS)

    Lauterio-Cruz, J. P.; Hernández-García, J. C.; Estudillo-Ayala, J. M.; Pottiez, O.; Rojas-Laguna, R.; Filoteo-Razo, J. D.; Samano-Aguilar, L. F.; Jauregui-Vazquez, D.

    2016-03-01

    In this work, we performed a numerical analysis of the supercontinuum spectrum generation in a couple of photonic crystal fibers with different structure. The proposed configuration initially has an input pulse with hyperbolic secant profile to generate noise-like pulses as output signal, by the Runge-Kutta method (RK4IP). By using the same configuration, now these noise-like pulses are used as pump for supercontinuum generation obtaining a broad and good flatness spectrum. The numerical analysis presented here demonstrates the potential of noise-like pulses from a passively mode-locked fiber laser for broadband spectrum generation combining two different photonic crystal fibers. Besides this paper helps to understand the phenomena of supercontinuum generation which is mainly related to Raman self-frequency shift.

  2. A porous silica rock ("tripoli") in the footwall of the Jurassic Úrkút manganese deposit, Hungary: composition, and origin through carbonate dissolution

    USGS Publications Warehouse

    Polgari, Marta; Szabo, Zoltan; Szabo-Drubina, Magda; Hein, James R.; Yeh, Hsueh-Wen

    2005-01-01

    The mineralogical, chemical, and isotopic compositions were determined for a white tripoli from the footwall of the Jurassic Úrkút Mn-oxide ore deposit in the Bakony Mountains, Hungary. The tripoli consists of quartz and chalcedony, with SiO2 contents up to 100 wt.%; consequently, trace-element contents are very low. Oxygen isotopes and quartz crystallinity indicate a low-temperature diagenetic origin for this deposit. The tripoli was formed by dissolution of the carbonate portion of the siliceous (sponge spicules) Isztimér Limestone. Dissolution of the carbonate was promoted by inorganic and organic acids generated during diagensis and left a framework composed of diagenetic silica that preserved the original volume of the limestone layer. The relative enrichment of silica and high porosity is the result of that carbonate dissolution. The silty texture of this highly friable rock is due to the structurally weak silica framework.

  3. Specific Anti-Leukemic Activity of the Peptide Warnericin RK and Analogues and Visualization of Their Effect on Cancer Cells by Chemical Raman Imaging.

    PubMed

    Loiseau, Clémence; Augenstreich, Jacques; Marchand, Adrienne; Harté, Etienne; Garcia, Martine; Verdon, Julien; Mesnil, Marc; Lecomte, Sophie; Berjeaud, Jean-Marc

    2016-01-01

    Antimicrobial peptides can be used as therapeutic agents against cancer cells. Warnericin RK and derivatives (WarnG20D and WarnF14V) were tested on various, solid tumor or leukemia, cancer cells. These peptides appeared to be cytotoxic on all the cell types tested, cancerous as well healthy, but very interestingly displayed no deleterious effect on healthy mononuclear cells. The mode of action of the peptide was proposed to be membranolytic, using chemical Raman imaging. Addition of peptide induced a large disorganization of the membrane leading to the loss of the content of inner compartments of Jurkat cell, whereas no effect was observed on the healthy mononuclear cells. The less hemolytic peptides WarnG20D and WarnF14V could be good candidates for the leukemia treatment. PMID:27598770

  4. Promoters of the Broad Host Range Plasmid Rk2: Analysis of Transcription (Initiation) in Five Species of Gram-Negative Bacteria

    PubMed Central

    Greener, A.; Lehman, S. M.; Helinski, D. R.

    1992-01-01

    A broad host range cloning vector was constructed, suitable for monitoring promoter activity in diverse Gram-negative bacteria. This vector, derived from plasmid RSF1010, utilized the firefly luciferase gene as the reporter, since the assay for its bioluminescent product is sensitive, and measurements can be made without background from the host. Twelve DNA fragments with promoter activity were obtained from broad host range plasmid RK2 and inserted into the RSF1010 derived vector. The relative luciferase activities were determined for these fragments in five species of Gram-negative bacteria. In addition, four promoters were analyzed by primer extension to locate transcriptional start sites in each host. The results show that several of the promoters vary substantially in relative strengths or utilize different transcriptional start sites in different bacteria. Other promoters exhibited similar activities and identical start sites in the five hosts examined. PMID:1732166

  5. The kil-kor regulon of broad-host-range plasmid RK2: nucleotide sequence, polypeptide product, and expression of regulatory gene korC.

    PubMed Central

    Kornacki, J A; Burlage, R S; Figurski, D H

    1990-01-01

    Broad-host-range plasmid RK2 encodes several kil operons (kilA, kilB, kilC, kilE) whose expression is potentially lethal to Escherichia coli host cells. The kil operons and the RK2 replication initiator gene (trfA) are coregulated by various combinations of kor genes (korA, korB, korC, korE). This regulatory network is called the kil-kor regulon. Presented here are studies on the structure, product, and expression of korC. Genetic mapping revealed the precise location of korC in a region near transposon Tn1. We determined the nucleotide sequence of this region and identified the korC structural gene by analysis of korC mutants. Sequence analysis predicts the korC product to be a polypeptide of 85 amino acids with a molecular mass of 9,150 daltons. The KorC polypeptide was identified in vivo by expressing wild-type and mutant korC alleles from a bacteriophage T7 RNA polymerase-dependent promoter. The predicted structure of KorC polypeptide has a net positive charge and a helix-turn-helix region similar to those of known DNA-binding proteins. These properties are consistent with the repressorlike function of KorC protein, and we discuss the evidence that KorA and KorC proteins act as corepressors in the control of the kilC and kilE operons. Finally, we show that korC is expressed from the bla promoters within the upstream transposon Tn1, suggesting that insertion of Tn1 interrupted a plasmid operon that may have originally included korC and kilC. Images PMID:2160936

  6. What do we know about the medical biography of Kemal Atatürk (1881-1938)? A summary of the state of knowledge and outlook on relevant issues for further research.

    PubMed

    Vatanoglu-Lutz, E Elif; Hot, Inci; Coban, Mustafa

    2013-08-01

    Mustafa Kemal Atatürk, the founder and first President of the Turkish republic, is the subject of many more or less 'heroic' biographies and few critical ones. His anamnesis, however, is only available in fragments. Many books omit details of Atatürk's health and life, for example his bloodline, his illness and eventually his death, his funeral prayer and ceremony and his burial. His liver problem, diagnosed as cirrhosis and said to be the cause of his death, is well described but his general health and other sicknesses are scarcely recorded. This paper provides an overview of his anamnesis as far as it is known, the literature describing it and the level of knowledge generally published, and it also indicates where original research in the archives is needed to complete the picture. PMID:24585761

  7. Solostamenides paucitesticulatus n. sp. (Monogenoidea: Mazocraeidea: Microcotylidae) from the freshwater mullet Liza abu (Heckel) (Mugiliformes: Mugilidae) from Atatürk Reservoir on the Euphrates River in southern Turkey.

    PubMed

    Kritsky, Delane C; Öktener, Ahmet

    2015-06-01

    Solostamenides paucitesticulatus n. sp. (Monogenoidea: Microcotylidae) from the gills of the abu mullet Liza abu (Heckel) in Atatürk Reservoir in southern Turkey is described. Among other features, the new species is easily distinguished from its three congeners, Solostamenides mugilis (Vogt, 1879), Solostamenides pseudomugilis (Hargis, 1956) and Solostamenides polyorchis Zhang & Yang, 2001, by having fewer hooks on the male copulatory organ (11 to 14), testes (5 to 9), and haptoral clamps (31 to 47). PMID:25962461

  8. The host range of RK2 minimal replicon copy-up mutants is limited by species-specific differences in the maximum tolerable copy number.

    PubMed

    Haugan, K; Karunakaran, P; Tøndervik, A; Valla, S

    1995-01-01

    The minimal replicon of the broad-host-range plasmid RK2 consists of a gene, trfA (trans-acting replication), encoding a protein required for initiation of plasmid replication. The TrfA protein binds to iterons in the cis-acting origin of vegetative replication (oriV), but the exact mechanism by which TrfA-mediated replication initiation takes place is not known. We report here the isolation and characterization of five mini RK2 trfA mutant plasmids with an elevated plasmid copy number, four in Pseudomonas aeruginosa and one in Azotobacter vinelandii. The mutations are localized between or downstream of previously reported Escherichia coli copy-up mutations in trfA, and one of the mutations has been described earlier as an independent copy-up isolate in E. coli. The five mutant plasmids were all moderately copy up in both E. coli and their host of origin, in spite of the use of isolation procedures which were expected to select efficiently in favor of plasmid mutants specifying high copy numbers. In contrast, previously described high copy-up mutants isolated in E. coli could not be established in P. aeruginosa and A. vinelandii. These high copy-up mutants were shown to induce cell killing in E. coli under conditions where the plasmid copy number was increased as a physiological response to reduced growth rate. We propose that the reason for this killing effect is that the copy number under these conditions exceeds an upper tolerance level specific for E. coli. By assuming that the corresponding tolerance level is lower in P. aeruginosa and A. vinelandii than in E. coli, and that the mechanism of copy number regulation is similar, the model can explain the phenotypes of all tested copy up mutants in these two hosts. Analogous studies were also performed in Salmonella typhimurium and Acetobacter xylinum. The data obtained in these studies indicate that the above model is probably generally true for gram-negative bacteria, and the results also indicate that the

  9. Mutation of the Myxoma virus SERP2 P1-site to prevent proteinase inhibition causes apoptosis in cultured RK-13 cells and attenuates disease in rabbits, but mutation to alter specificity causes apoptosis without reducing virulence.

    PubMed

    MacNeill, Amy L; Turner, Peter C; Moyer, Richard W

    Myxoma virus (MYX) prevents apoptosis in RK-13 cells and forms thick dermal lesions with 100% mortality in rabbits. MYX encodes the virulence factor SERP2, a serine proteinase inhibitor (serpin). SERP2 was mutated to evaluate SERP2 function during MYX infection. MYXDeltaSERP2::lacZ (deleted for SERP2) did not inhibit apoptosis in RK-13 cells; infected rabbits had thin dermal lesions and <10% mortality. MYX-SERP2-D294A, a P1-site aspartate to alanine mutant, inactivated the serpin; infection was indistinguishable from MYXDeltaSERP2::lacZ. SERP2-D294E prevented inhibition of caspase-8, caspase-10 and granzyme-B; and MYX-SERP2-D294E failed to block apoptosis in RK-13 cells, but was fully virulent in rabbits. MYXDeltaSERP2::crmA expressed crmA instead of SERP2 and inhibited apoptosis in cell culture, but caused thin lesions and only 70% mortality in rabbits, hence crmA cannot fully substitute for SERP2. Control of apoptosis in culture does not correlate with virulence in rabbits. Virulence may instead depend on inhibition of proinflammatory proteinases by SERP2. PMID:16959285

  10. Antisense SNF1-related (SnRK1) protein kinase gene represses transient activity of an alpha-amylase (alpha-Amy2) gene promoter in cultured wheat embryos.

    PubMed

    Laurie, Sophie; McKibbin, Rowan S; Halford, Nigel G

    2003-02-01

    A DNA fragment corresponding to part of an SNF1 (sucrose non-fermenting-1)-related protein kinase (SnRK1) transcript was amplified by a polymerase chain reaction (PCR) from a wheat (Triticum aestivum) endosperm cDNA library. It was used to construct a chimaeric gene, pUasSnRKN, comprising a ubiquitin promoter, the SnRK1 PCR product in the antisense orientation and the nopaline synthase (Nos) gene terminator. This construct was used in transient gene expression experiments in cultured wheat embryos together with a series of reporter gene constructs. These included the wheat alpha amylase gene alpha-Amy2 promoter with UidA (Gus) coding region (palpha2GT), rice actin promoter with Gus (pActIDGus), ubiquitin promoter with Gus (pAHC25) and actin promoter with green fluorescent protein (GFP) gene (pAct1Is-GFP1). All of the reporter genes were found to be active when bombarded into scutellae isolated from immature grains at 25 d post-anthesis and incubated on MS medium for 24 h prior to bombardment. However, co-bombardment of palpha2GT with equimolar amounts of pUasSnRKN resulted in no detectable Gus activity, indicating that the antisense SnRK1 construct repressed the alpha-Amy2 promoter. Co-bombardment with pUasSnRKN had no effect on the activity of the other promoters used in the study. A triple bombardment with palpha2GT, pAct1Is-GFP-1 and pUasSnRKN resulted in clear green fluorescence, indicating that the bombarded cells were still viable, but no Gus activity. RT-PCR analysis showed clearly that the antisense SnRK1 gene was expressing. Northern and RT-PCR analyses confirmed that SnRK1 and both alpha-amylase genes, alpha-Amy1 and alpha-Amy2, are expressed in cultured wheat embryos harvested from grain 25 d post-anthesis. Expression of alpha-Amy1 and alpha-Amy2 was up-regulated by sugar starvation. PMID:12554717

  11. Suppressor of K+ transport growth defect 1 (SKD1) interacts with RING-type ubiquitin ligase and sucrose non-fermenting 1-related protein kinase (SnRK1) in the halophyte ice plant.

    PubMed

    Chiang, Chih-Pin; Li, Chang-Hua; Jou, Yingtzy; Chen, Yu-Chan; Lin, Ya-Chung; Yang, Fang-Yu; Huang, Nu-Chuan; Yen, Hungchen Emilie

    2013-05-01

    SKD1 (suppressor of K+ transport growth defect 1) is an AAA-type ATPase that functions as a molecular motor. It was previously shown that SKD1 accumulates in epidermal bladder cells of the halophyte Mesembryanthemum crystallinum. SKD1 knock-down Arabidopsis mutants showed an imbalanced Na+/K+ ratio under salt stress. Two enzymes involved in protein post-translational modifications that physically interacted with McSKD1 were identified. McCPN1 (copine 1), a RING-type ubiquitin ligase, has an N-terminal myristoylation site that links to the plasma membrane, a central copine domain that interacts with McSKD1, and a C-terminal RING domain that catalyses protein ubiquitination. In vitro ubiquitination assay demonstrated that McCPN1 was capable of mediating ubiquitination of McSKD1. McSnRK1 (sucrose non-fermenting 1-related protein kinase) is a Ser/Thr protein kinase that contains an N-terminal STKc catalytic domain to phosphorylate McSKD1, and C-terminal UBA and KA1 domains to interact with McSKD1. The transcript and protein levels of McSnRK1 increased as NaCl concentrations increased. The formation of an SKD1-SnRK1-CPN1 ternary complex was demonstrated by yeast three-hybrid and bimolecular fluorescence complementation. It was found that McSKD1 preferentially interacts with McSnRK1 in the cytosol, and salt induced the re-distribution of McSKD1 and McSnRK1 towards the plasma membrane via the microtubule cytoskeleton and subsequently interacted with RING-type E3 McCPN1. The potential effects of ubiquitination and phosphorylation on McSKD1, such as changes in the ATPase activity and cellular localization, and how they relate to the functions of SKD1 in the maintenance of Na+/K+ homeostasis under salt stress, are discussed. PMID:23580756

  12. Inductive measurements of the stress assisted and strain induced martensite transformations of Sandvik maraging steel 1RK91 before, during and after metal forming

    NASA Astrophysics Data System (ADS)

    Nolles, H.; Post, J.; Beyer, J.

    2003-10-01

    Measurements of transformation behaviour are very important to get a good knowledge of the material behaviour of metastable stainless steels during metal forming processes. A convenient way of measuring this kind of transformations is inductive measurment. This article describes a possible solution for this kind of measurements, together with the developed sensors and electronics. The first part of the article focuses on the electronics, calibration and the development of the sensors. The equipment is also used for multi sensor measurements. The second part of the article shows results of measurements on the Sandvik maraging steel 1RK91. The results are shown for three examples: A problem that occurs during inductive measurements is that more pararneters than the martensite content have influence on the output of the signal. Elastic stresses, temperature and plastic deformation have also a big influence on the signal output. Some results of measurements are shown to demonstrate the influence together with a calibration metho(l.Thieineasuring device is fully automatic and conriceted to a computer to make sample logging possible during testing.

  13. Multi-centric prospective evaluation of rk39 rapid test and direct agglutination test for the diagnosis of visceral leishmaniasis in Brazil.

    PubMed

    de Assis, Tália S M; Braga, Alexandre S da C; Pedras, Mariana J; Oliveira, Edward; Barral, Aldina; de Siqueira, Isadora C; Costa, Carlos H N; Costa, Dorcas L; Holanda, Thiago A; Soares, Vítor Y R; Biá, Mauro; Caldas, Arlene de J M; Romero, Gustavo A S; Rabello, Ana

    2011-02-01

    The diagnosis of visceral leishmaniasis (VL) is still a major problem in Brazil and several other countries where the disease is endemic. The use of an easy-to-use and interpret, sensitive, and specific method that requires no complex infrastructure or specialized professionals, such as direct agglutination test (DAT) and the rK39-based rapid immunochromatographic test may enhance the diagnosis of disease. This study evaluated the performance of a rapid test (DiaMed- IT-LEISH®) and the DAT for the diagnosis of VL in 213 parasitologically confirmed cases and 119 controls with clinical suspicion of VL and confirmation of another etiology. The sensitivities and specificities of the rapid test were 93% and 97%, respectively and those of the DAT were 90% and 96%, respectively. The positive predictive values of the rapid test and the DAT were 98% and 97%, respectively and the negative predictive values were 89% and 84%, respectively. The Kappa index showed agreement between both methods classified as substantial (0.77). This study showed that the DAT and the rapid test can be used to diagnose VL in Brazil, following a pilot study for implementation of the rapid test in the health services. PMID:20970152

  14. Youths' perception and knowledge towards environmental problems in a developing country: in the case of Atatürk University, Turkey.

    PubMed

    Kulözü, Neslihan

    2016-06-01

    This study explores the perception and knowledge of environmental problems among students of Atatürk University, Turkey, as a developing country, by comparing their perceptions with known facts about environmental problems at local, national, and global levels. In addition, the correlation between the student groups and their knowledge levels and the differences between the student groups according to their knowledge levels when taking age and gender into account are explored. To this end, the research designed as a case study and pursued an exploratory approach revealed that major perceived environmental problems show differences at local, national, and global levels; that consensus on both major perceived problems and knowledge decreases from a local to global level; that a weak correlation exists between the age (r (205) = -.010; .002; -.071) and gender groups (r (205) = -.099; -.125; .065) in terms of their knowledge at local, national, and global level, respectively; and that the contribution of knowledge at a national level can help to explain differences among the age groups (-.304; .102; -.148) at national level and the gender groups (.131; -.205) at national and global levels. PMID:27117145

  15. TrfA-Dependent Inner Membrane-Associated Plasmid RK2 DNA Synthesis and Association of TrfA with Membranes of Different Gram-Negative Hosts

    PubMed Central

    Banack, Trevor; Kim, Peter D.; Firshein, William

    2000-01-01

    TrfA, the replication initiator protein of broad-host-range plasmid RK2, was tested for its ability to bind to the membrane of four different gram-negative hosts in addition to Escherichia coli: Pseudomonas aeruginosa, Pseudomonas putida, Salmonella enterica serovar Typhimurium, and Rhodobacter sphaeroides. Cells harboring TrfA-encoding plasmids were fractionated into soluble, inner membrane, and outer membrane fractions. The fractions were subjected to Western blotting, and the blots were probed with antibody to the TrfA proteins. TrfA was found to fractionate with the cell membranes of all species tested. When the two membrane fractions of these species were tested for their ability to synthesize plasmid DNA endogenously (i.e., without added template or enzymes), only the inner membrane fraction was capable of extensive synthesis that was inhibited by anti-TrfA antibody in a manner similar to that of the original host species, E. coli. In addition, although DNA synthesis did occur in the outer membrane fraction, it was much less extensive than that exhibited by the inner membrane fraction and only slightly affected by anti-TrfA antibody. Plasmid DNA synthesized by the inner membrane fraction of one representative species, P. aeruginosa, was characteristic of supercoil and intermediate forms of the plasmid. Extensive DNA synthesis was observed in the soluble fraction of another representative species, R. sphaeroides, but it was completely unaffected by anti-TrfA antibody, suggesting that such synthesis was due to repair and/or nonspecific chain extension of plasmid DNA fragments. PMID:10913068

  16. Geodetic activities of the Department of Defense under IGY programs

    SciTech Connect

    Williams, O.W.; Daugherty, K.I.

    1983-10-16

    Attention is given to the U.S. Department of Defence (DOD) activities that contributed to the International Geophysical Year's active, passive, and cooperative satellite programs. The DOD continues to support the deployment, enhancement, and application of novel technology in such areas as satellite altimetry, gravity radiometry, inertial surveying, interferometry, airborne gravimetry, inertial surveying, and CCD and laser methods for geodetic astronomy. Also noted are such major department initiatives as the Global Positioning System, which will become operational toward the end of this decade.

  17. A link between magnesium-chelatase H subunit and sucrose nonfermenting 1 (SNF1)-related protein kinase SnRK2.6/OST1 in Arabidopsis guard cell signalling in response to abscisic acid

    PubMed Central

    Liang, Shan; Lu, Kai; Wu, Zhen; Jiang, Shang-Chuan; Yu, Yong-Tao; Bi, Chao; Xin, Qi; Wang, Xiao-Fang; Zhang, Da-Peng

    2015-01-01

    Magnesium-chelatase H subunit [CHLH/putative abscisic acid (ABA) receptor ABAR] positively regulates guard cell signalling in response to ABA, but the molecular mechanism remains largely unknown. A member of the sucrose nonfermenting 1 (SNF1)-related protein kinase 2 family, SnRK2.6/open stomata 1 (OST1)/SRK2E, which plays a critical role in ABA signalling in Arabidopsis guard cells, interacts with ABAR/CHLH. Neither mutation nor over-expression of the ABAR gene affects significantly ABA-insensitive phenotypes of stomatal movement in the OST1 knockout mutant allele srk2e. However, OST1 over-expression suppresses ABA-insensitive phenotypes of the ABAR mutant allele cch in stomatal movement. These genetic data support that OST1 functions downstream of ABAR in ABA signalling in guard cells. Consistent with this, ABAR protein is phosphorylated, but independently of the OST1 protein kinase. Two ABAR mutant alleles, cch and rtl1, show ABA insensitivity in ABA-induced reactive oxygen species and nitric oxide production, as well as in ABA-activated phosphorylation of a K+ inward channel KAT1 in guard cells, which is consistent with that observed in the pyr1 pyl1 pyl2 pyl4 quadruple mutant of the well-characterized ABA receptor PYR/PYL/RCAR family acting upstream of OST1. These findings suggest that ABAR shares, at least in part, downstream signalling components with PYR/PYL/RCAR receptors for ABA in guard cells; though cch and rtl1 show strong ABA-insensitive phenotypes in both ABA-induced stomatal closure and inhibition of stomatal opening, while the pyr1 pyl1 pyl2 pyl4 quadruple mutant shows strong ABA insensitivity only in ABA-induced stomatal closure. These data establish a link between ABAR/CHLH and SnRK2.6/OST1 in guard cell signalling in response to ABA. PMID:26175350

  18. A link between magnesium-chelatase H subunit and sucrose nonfermenting 1 (SNF1)-related protein kinase SnRK2.6/OST1 in Arabidopsis guard cell signalling in response to abscisic acid.

    PubMed

    Liang, Shan; Lu, Kai; Wu, Zhen; Jiang, Shang-Chuan; Yu, Yong-Tao; Bi, Chao; Xin, Qi; Wang, Xiao-Fang; Zhang, Da-Peng

    2015-10-01

    Magnesium-chelatase H subunit [CHLH/putative abscisic acid (ABA) receptor ABAR] positively regulates guard cell signalling in response to ABA, but the molecular mechanism remains largely unknown. A member of the sucrose nonfermenting 1 (SNF1)-related protein kinase 2 family, SnRK2.6/open stomata 1 (OST1)/SRK2E, which plays a critical role in ABA signalling in Arabidopsis guard cells, interacts with ABAR/CHLH. Neither mutation nor over-expression of the ABAR gene affects significantly ABA-insensitive phenotypes of stomatal movement in the OST1 knockout mutant allele srk2e. However, OST1 over-expression suppresses ABA-insensitive phenotypes of the ABAR mutant allele cch in stomatal movement. These genetic data support that OST1 functions downstream of ABAR in ABA signalling in guard cells. Consistent with this, ABAR protein is phosphorylated, but independently of the OST1 protein kinase. Two ABAR mutant alleles, cch and rtl1, show ABA insensitivity in ABA-induced reactive oxygen species and nitric oxide production, as well as in ABA-activated phosphorylation of a K(+) inward channel KAT1 in guard cells, which is consistent with that observed in the pyr1 pyl1 pyl2 pyl4 quadruple mutant of the well-characterized ABA receptor PYR/PYL/RCAR family acting upstream of OST1. These findings suggest that ABAR shares, at least in part, downstream signalling components with PYR/PYL/RCAR receptors for ABA in guard cells; though cch and rtl1 show strong ABA-insensitive phenotypes in both ABA-induced stomatal closure and inhibition of stomatal opening, while the pyr1 pyl1 pyl2 pyl4 quadruple mutant shows strong ABA insensitivity only in ABA-induced stomatal closure. These data establish a link between ABAR/CHLH and SnRK2.6/OST1 in guard cell signalling in response to ABA. PMID:26175350

  19. Usefulness of the rK39-Immunochromatographic Test, Direct Agglutination Test, and Leishmanin Skin Test for Detecting Asymptomatic Leishmania Infection in Children in a New Visceral Leishmaniasis Focus in Amhara State, Ethiopia

    PubMed Central

    Gadisa, Endalamaw; Custodio, Estefanía; Cañavate, Carmen; Sordo, Luis; Abebe, Zelalem; Nieto, Javier; Chicharro, Carmen; Aseffa, Abraham; Yamuah, Lawrence; Engers, Howard; Moreno, Javier; Cruz, Israel

    2012-01-01

    In areas where visceral leishmaniasis is anthroponotic, asymptomatically infected patients may play a role in transmission. Additionally, the number of asymptomatic patients in a disease-endemic area will also provide information on transmission dynamics. Libo Kemkem and Fogera districts (Amhara State, Ethiopia) are now considered newly established areas to which visceral leishmaniasis is endemic. In selected villages in these districts, we conducted a study to assess the usefulness of different approaches to estimate the asymptomatic infection rate. Of 605 participants, the rK39 immunochromatographic test was able to detect asymptomatic infection in 1.5% (9 of 605), direct agglutination test in 5.3% (32 of 605), and leishmanin skin test in 5.6% (33 of 589); the combined use of serologic methods and leishmanin skin test enabled detecting asymptomatic infection in 10.1% (61 of 605). We conclude that the best option to detect asymptomatic infection in this new visceral leishmaniasis–endemic focus is the combined use of the direct agglutination test and the leishmanin skin test. PMID:22556076

  20. The anti-toxin ParD of plasmid RK2 consists of two structurally distinct moieties and belongs to the ribbon-helix-helix family of DNA-binding proteins.

    PubMed Central

    Oberer, Monika; Zangger, Klaus; Prytulla, Stefan; Keller, Walter

    2002-01-01

    NMR and CD spectroscopy have been used to characterize, both structurally and dynamically, the 82-amino-acid ParD protein of the post-segregational killing module of the broad-host-range plasmid RP4/RK2. ParD occurs as a dimer in solution and exercises two different control functions; an autoregulatory function by binding to its own promoter P(parDE) and a plasmid-stabilizing function by inhibiting ParE toxicity in cells that express ParD and ParE. Analysis of the secondary structure based on the chemical-shift indices, sequential nuclear Overhauser enhancements (NOEs) and (3)J(Halpha-NH) scalar coupling constants showed that the N-terminal domain of ParD consists of a short beta-ribbon followed by three alpha-helices, demonstrating that ParD contains a ribbon-helix-helix fold, a DNA-binding motif found in a family of small prokaryotic repressors. (15)N longitudinal (T(1)) and transverse (T(2)) relaxation measurements and hetero nuclear NOEs showed that ParD is divided into two separate domains, a well-ordered N-terminal domain and a very flexible C-terminal domain. An increase in secondary structure was observed upon addition of trifluoroethanol, suggested to result from the formation of structured stretches in the C-terminal part of the protein. This is the first experimental evidence that the DNA-binding domain of ParD belongs to the ribbon-helix-helix fold family, and this structural motif is proposed to be present in functionally similar antidote proteins. PMID:11743881

  1. Activation of stress-activated protein kinase-3 (SAPK3) by cytokines and cellular stresses is mediated via SAPKK3 (MKK6); comparison of the specificities of SAPK3 and SAPK2 (RK/p38).

    PubMed Central

    Cuenda, A; Cohen, P; Buée-Scherrer, V; Goedert, M

    1997-01-01

    Stress-activated protein kinase-3 (SAPK3), a recently described MAP kinase family member with a wide-spread tissue distribution, was transfected into several mammalian cell lines and shown to be activated in response to cellular stresses, interleukin-1 (IL-1) and tumour necrosis factor (TNF) in a similar manner to SAPK1 (also termed JNK) and SAPK2 (also termed p38, RK, CSBP and Mxi2). SAPK3 and SAPK2 were activated at similar rates in vitro by SAPKK3 (also termed MKK6), and SAPKK3 was the only activator of SAPK3 that was induced when KB or 293 cells were exposed to cellular stresses or stimulated with IL-1 or TNF. Co-transfection with SAPKK3 induced SAPK3 activity and greatly enhanced activation in response to osmotic shock. These experiments indicate that SAPKK3 mediates the activation of SAPK3 in several mammalian cells. SAPK3 and SAPK2 phosphorylated a number of proteins at similar rates, including the transcription factors ATF2, Elk-1 and SAP1, but SAPK3 was far less effective than SAPK2 in activating MAPKAP kinase-2 and MAPKAP kinase-3. Unlike SAPK2, SAPK3 was not inhibited by the drug SB 203580. SAPK3 phosphorylated ATF2 at Thr69, Thr71 and Ser90, the same residues phosphorylated by SAPK1, whereas SAPK2 only phosphorylated Thr69 and Thr71. Our results suggest that cellular functions previously attributed to SAPK1 and/or SAPK2 may be mediated by SAPK3. PMID:9029150

  2. On the standard model predictions for R_K and R_{K^*}

    NASA Astrophysics Data System (ADS)

    Bordone, Marzia; Isidori, Gino; Pattori, Andrea

    2016-08-01

    We evaluate the impact of radiative corrections in the ratios Γ [B→ M μ ^+μ ^-]/Γ [B→ M e^+e^-] when the meson M is a K or a K^*. Employing the cuts on m^2_{ℓ ℓ } and the reconstructed B-meson mass presently applied by the LHCb Collaboration, such corrections do not exceed a few %. Moreover, their effect is well described (and corrected for) by existing Monte Carlo codes. Our analysis reinforces the interest of these observables as clean probe of physics beyond the Standard Model.

  3. Integrated geographic information systems (IGIS) analysis and definition of the tectonic framework of northern Mexico

    NASA Astrophysics Data System (ADS)

    Martinez Pina, Carlos Manuel

    Crustal rupture structures reactivated in the course of the tectonic history of northern Mexico are the surface expressions of planes of weakness, in the form of simple or composite rectilinear features or slightly curved, defined as lineaments. Unless otherwise defined as strike-slip faults, lineaments are part of parallel and sub-parallel oblique convergent or oblique divergent tectonic zones cross cutting the Sierra Madre Occidental and northern Mexico, in a NW trend. These shear zones are the response to the oblique subduction of the Farallon plate beneath North America. Kinematic analysis of five selected sites in northern Mexico, three basins and two compressional shear zones, proved possible a combination of shear mechanism diagram and models from analogue materials, with satellite imagery and geographic information systems, as an aid to define strike-slip fault motion. This was done using a reverse engineering process by comparing geometries. One of the sites assessed, involving the Parras Basin, Coahuila Block (CB), San Marcos fault, a postulated PBF-1 fault, allowed for palinpastic reconstruction of the CB that corroborated the results of the vector motion defined, in addition to an extension of ˜25% in a northwest southeast direction. A GIS-based compilation and georeferenced regional structural studies by several researchers were used as ground control areas (GCA); their interpolation and interpretation, resulted in a tectonic framework map of northern Mexico. In addition, shaded relief models overlaid by the lineaments / fault layer allowed structural analyses of basins related to these major structures. Two important results were obtained from this study: the Tepehuanes-San Luis-fault (TSL) and the Guadalupe fault, named herein, displaces the Villa de Reyes graben, and the Aguascalientes graben, respectively, to the SE, confirming their left lateral vector motion; afterwards TSL was displaced south by the right lateral strike slip Taxco-San Miguel de Allende fault. The second result refers to the hypothesis that the Mesa Central was brought to its present location by a subduction zone located to the north. This subduction zone coincides with several researchers who postulated the idea. The compressional zones refer to segments of the Sinforosa and a postulated Aquinquari fault located in the stratotectonic Guerrero Terrane regarded as a highly mineralized zone. Negative anomalies near -200 milligals are strongly suggestive of a cratonic block identified in western Chihuahua, it being named the Western Chihuahua Cratonic Block (WCCB). In the southwestern portion of the North American craton the age provinces are well documented, but the block versus mobile belt idea has not been put forth or emphasized. The present study combines data of several types, sedimentological, structural, igneous geochemistry, and geochronologic data to evaluate this behavior in SW NA, and the proposed block is tested against these data. The presence of the WCCB is supported by a wide variety of data. Basins, troughs, aulacogens, bimodal volcanism, and other rift and rift shoulder features, characterize the spatially constrained mobile belts. Mobile belts surrounding the WCCB contain geologic records of the events going back to 1.4 Ga, with different aspects being dominant over geologic time. Mobile belts will participate in compression,(subduction), extension (rifting), and transform (lateral) faulting. The WCCB may have been derived from closely, adjacent, North American craton by mobile belt action. This study has shown that integration of data is essential, because allows detection of differences in hypotheses for the same event in the same area. This integration capability is what makes integrated geographic information systems a powerful tool, not only for their synergy, but because they can be combined with specific techniques that provide data before going to conduct fieldwork. Whether the issue of defining the tectonic framework of northern Mexico can be resolved or not, depends on the viability of integrating volumes of data from research, hypotheses, or maps, and put together under the same geographic frame.

  4. [Translation and comments of the first three tablets of the IGI series].

    PubMed

    Attia, Annie

    2015-01-01

    This paper translates the chapter on eye diseases from the great treatise on therapeutics compiled under Ashurbanipal and found in his royal libraries at Nineveh. As a professional ophthalmologist, I try to analyse and to understand the symptoms and the treatments described in these tablets, while bearing in mind the dangers of retrospective diagnosis. PMID:26827500

  5. Protection against Coccidiosis: Role of Passive Immunity (Immediate Immunity) Mediated by Hyperimmune IgY Antibodies

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Avian coccidiosis is a major parasitic disease of poultry with substantial economic consequences. Although good management practices and live vaccination with prophylactic medication have helped to reduce the spread of coccidiosis, alternative control strategies need to be developed due to the evid...

  6. Protection Against Coccidiosis: Role of passive immunity (immediate immunity) mediated by hyperimmune IgY antibodies

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Avian coccidiosis is a major parasitic disease of poultry with substantial economic consequences estimated to cost the industry greater than $3 billion in annual losses worldwide. Coccidiosis is caused by several distinct species of the genus Eimeria that infect the gut and are transmitted between ...

  7. Generation and characterization of chicken egg yolk antibodies (IgY) against TNFR1.

    PubMed

    Hashemi, M; Amirijavid, S; Entezari, M; Shafaroodi, H; Saghafi, Z Jokar

    2015-01-01

    TNF is from a big family of cytokines with different activities in different parts of the body. Among the various activities of TNFR1, induction of apoptosis by a receptor appears to be an attractive and promising one. This can be achieved through the death domain of the receptor in cells that are stimulated by ligand, to induce apoptosis. Activation of the receptor occurs through its occupation by ligands or its antagonists such as antibodies. Several kinds of antibodies, including antibodies of mammals and birds are used in the research and therapy field. Avian antibodies are highly regarded which is due to the many positive characteristics they have. Firstly, total protein of TNFR1 was cloned. Blood sampling was performed, white blood cell separation, extraction of RNA and at cDNA synthesis. After making sure from synthesis of cDNA, it was used as template for PCR reaction. The cloned fragment in the prokaryotic expression vector, pET28a, transferred to prokaryotic host, BL21(DE3) and the protein (TNFR1) expressed. After protein purification by affinity column were injected to immunize the chickens. Interestingly, antibodies purified from egg yolk of immunized chickens, in ELISA assay showed sufficient specificity. Such antibodies could able to ensure quick and immediate protection against several biotargets (Fig. 4, Ref. 37). PMID:25924641

  8. Test report : Raytheon / KTech RK30 energy storage system.

    SciTech Connect

    Rose, David Martin; Schenkman, Benjamin L.; Borneo, Daniel R.

    2013-10-01

    The Department of Energy Office of Electricity (DOE/OE), Sandia National Laboratories (SNL) and the Base Camp Integration Lab (BCIL) partnered together to incorporate an energy storage system into a microgrid configured Forward Operating Base to reduce the fossil fuel consumption and to ultimately save lives. Energy storage vendors will be sending their systems to SNL Energy Storage Test Pad (ESTP) for functional testing and then to the BCIL for performance evaluation. The technologies that will be tested are electro-chemical energy storage systems comprising of lead acid, lithium-ion or zinc-bromide. Raytheon/KTech has developed an energy storage system that utilizes zinc-bromide flow batteries to save fuel on a military microgrid. This report contains the testing results and some limited analysis of performance of the Raytheon/KTech Zinc-Bromide Energy Storage System.

  9. A dominant gene for garnet brown seed coats at the Rk locus in Dorado common bean and mapping Rk to linkage group 1

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The color of the seed coats of Dorado (Phaseolus vulgaris L.) is garnet brown (dark red kidney bean color) and differs from most other dry bean varieties in the Honduran red bean market class. A genetic investigation of the color of Dorado (same as DOR364) and G19833 (Liborino market class) see...

  10. A dominant gene for garnet brown seed coats at the Rk locus in Dorado common bean and mapping Rk to linkage group 1

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The color of the seed coats of Dorado (Phaseolus vulgaris L.) is garnet brown (dark red kidney bean color) and differs from most other dry bean varieties in the Honduran red bean market class. A genetic investigation of the color of Dorado (same as DOR364) and G19833 (Liborino market class) seed co...

  11. Microscale Depletion of High Abundance Proteins in Human Biofluids using IgY14 Immunoaffinity Resin. Analysis of Human Plasma and Cerebrospinal Fluid

    SciTech Connect

    Hyung, Seok Won; Piehowski, Paul D.; Moore, Ronald J.; Orton, Daniel J.; Schepmoes, Athena A.; Clauss, Therese RW; Chu, Rosalie K.; Fillmore, Thomas L.; Brewer, Heather M.; Liu, Tao; Zhao, Rui; Smith, Richard D.

    2014-09-06

    Removal of highly abundant proteins in plasma is often carried out using immunoaffinity depletion to extend the dynamic range of measurements to lower abundance species. While commercial depletion columns are available for this purpose, they generally are not applicable to limited sample quantities (<20 µL) due to low yields stemming from losses caused by nonspecific binding to the column matrix. Additionally, the cost of the depletion media can be prohibitive for larger scale studies. Modern LC-MS instrumentation provides the sensitivity necessary to scale-down depletion methods with minimal sacrifice to proteome coverage, which makes smaller volume depletion columns desirable for maximizing sample recovery when samples are limited, as well as for reducing the expense of large scale studies. We characterized the performance of a 346 µL column volume micro-scale depletion system, using four different flow rates to determine the most effective depletion conditions for ~6 μL injections of human plasma proteins and then evaluated depletion reproducibility at the optimum flow rate condition. Depletion of plasma using a commercial 10 mL depletion column served as the control. Results showed depletion efficiency of the micro-scale column increased as flow rate decreased, and that our micro-depletion was reproducible. In an initial application, a 600 µL sample of human cerebral spinal fluid (CSF) pooled from multiple sclerosis patients was depleted and then analyzed using reversed phase liquid chromatography-mass spectrometry to demonstrate the utility of the system for this important biofluid where sample quantities are more commonly limited.

  12. Multiple IgH Isotypes Including IgD, Subclasses of IgM, and IgY Are Expressed in the Common Ancestors of Modern Birds.

    PubMed

    Han, Binyue; Yuan, Hui; Wang, Tao; Li, Bo; Ma, Li; Yu, Shuyang; Huang, Tian; Li, Yan; Fang, Dongming; Chen, Xiaoli; Wang, Yongsi; Qiu, Si; Guo, Ying; Fei, Jing; Ren, Liming; Pan-Hammarström, Qiang; Hammarström, Lennart; Wang, Jun; Wang, Jian; Hou, Yong; Pan, Qingjie; Xu, Xun; Zhao, Yaofeng

    2016-06-15

    Although evolutionarily just as ancient as IgM, it has been thought for many years that IgD is not present in birds. Based on the recently sequenced genomes of 48 bird species as well as high-throughput transcriptome sequencing of immune-related tissues, we demonstrate in this work that the ostrich (Struthio camelus) possesses a functional δ gene that encodes a membrane-bound IgD H chain with seven CH domains. Furthermore, δ sequences were clearly identified in many other bird species, demonstrating that the δ gene is widely distributed among birds and is only absent in certain bird species. We also show that the ostrich possesses two μ genes (μ1, μ2) and two υ genes (υ1, υ2), in addition to the δ and α genes. Phylogenetic analyses suggest that subclass diversification of both the μ and υ genes occurred during the early stages of bird evolution, after their divergence from nonavian reptiles. Although the positions of the two υ genes are unknown, physical mapping showed that the remaining genes are organized in the order μ1-δ-α-μ2, with the α gene being inverted relative to the others. Together with previous studies, our data suggest that birds and nonavian reptile species most likely shared a common ancestral IgH gene locus containing a δ gene and an inverted α gene. The δ gene was then evolutionarily lost in selected birds, whereas the α gene lost in selected nonavian reptiles. The data obtained in this study provide significant insights into the understanding of IgH gene evolution in tetrapods. PMID:27183632

  13. Microscale depletion of high abundance proteins in human biofluids using IgY14 immunoaffinity resin: Analysis of human plasma and cerebrospinal fluid

    SciTech Connect

    Hyung, Seok Won; Piehowski, Paul D.; Moore, Ronald J.; Orton, Daniel J.; Schepmoes, Athena A.; Clauss, Therese R.; Chu, Rosalie K.; Fillmore, Thomas L.; Brewer, Heather M.; Liu, Tao; Zhao, Rui; Smith, Richard D.

    2014-09-06

    Removal of highly abundant proteins in plasma is often carried out using immunoaffinity depletion to extend the dynamic range of measurements to lower abundance species. While commercial depletion columns are available for this purpose, they generally are not applicable to limited sample quantities (<20 µL) due to low yields stemming from losses caused by nonspecific binding to the column matrix. Additionally, the cost of the depletion media can be prohibitive for larger scale studies. Modern LC-MS instrumentation provides the sensitivity necessary to scale-down depletion methods with minimal sacrifice to proteome coverage, which makes smaller volume depletion columns desirable for maximizing sample recovery when samples are limited, as well as for reducing the expense of large scale studies. We characterized the performance of a 346 µL column volume micro-scale depletion system, using four different flow rates to determine the most effective depletion conditions for ~6 μL injections of human plasma proteins and then evaluated depletion reproducibility at the optimum flow rate condition. Depletion of plasma using a commercial 10 mL depletion column served as the control. Results showed depletion efficiency of the micro-scale column increased as flow rate decreased, and that our micro-depletion was reproducible. We found, in an initial application, a 600 µL sample of human cerebral spinal fluid (CSF) pooled from multiple sclerosis patients was depleted and then analyzed using reversed phase liquid chromatography-mass spectrometry to demonstrate the utility of the system for this important biofluid where sample quantities are more commonly limited.

  14. Microscale depletion of high abundance proteins in human biofluids using IgY14 immunoaffinity resin: Analysis of human plasma and cerebrospinal fluid

    DOE PAGESBeta

    Hyung, Seok Won; Piehowski, Paul D.; Moore, Ronald J.; Orton, Daniel J.; Schepmoes, Athena A.; Clauss, Therese R.; Chu, Rosalie K.; Fillmore, Thomas L.; Brewer, Heather M.; Liu, Tao; et al

    2014-09-06

    Removal of highly abundant proteins in plasma is often carried out using immunoaffinity depletion to extend the dynamic range of measurements to lower abundance species. While commercial depletion columns are available for this purpose, they generally are not applicable to limited sample quantities (<20 µL) due to low yields stemming from losses caused by nonspecific binding to the column matrix. Additionally, the cost of the depletion media can be prohibitive for larger scale studies. Modern LC-MS instrumentation provides the sensitivity necessary to scale-down depletion methods with minimal sacrifice to proteome coverage, which makes smaller volume depletion columns desirable for maximizingmore » sample recovery when samples are limited, as well as for reducing the expense of large scale studies. We characterized the performance of a 346 µL column volume micro-scale depletion system, using four different flow rates to determine the most effective depletion conditions for ~6 μL injections of human plasma proteins and then evaluated depletion reproducibility at the optimum flow rate condition. Depletion of plasma using a commercial 10 mL depletion column served as the control. Results showed depletion efficiency of the micro-scale column increased as flow rate decreased, and that our micro-depletion was reproducible. We found, in an initial application, a 600 µL sample of human cerebral spinal fluid (CSF) pooled from multiple sclerosis patients was depleted and then analyzed using reversed phase liquid chromatography-mass spectrometry to demonstrate the utility of the system for this important biofluid where sample quantities are more commonly limited.« less

  15. Microscale depletion of high abundance proteins in human biofluids using IgY14 immunoaffinity resin: analysis of human plasma and cerebrospinal fluid

    PubMed Central

    Hyung, Seok-Won; Piehowski, Paul D.; Moore, Ronald J.; Orton, Daniel J.; Schepmoes, Athena A.; Clauss, Therese R.; Chu, Rosalie K.; Fillmore, Thomas L.; Brewer, Heather; Liu, Tao; Zhao, Rui; Smith, Richard D.

    2014-01-01

    Removal of highly abundant proteins in plasma is often carried out using immunoaffinity depletion to extend the dynamic range of measurements to lower abundance species. While commercial depletion columns are available for this purpose, they generally are not applicable to limited sample quantities (<20 μL) due to low yields stemming from losses caused by nonspecific binding to the column matrix and concentration of large eluent volumes. Additionally, the cost of the depletion media can be prohibitive for larger-scale studies. Modern LC-MS instrumentation provides the sensitivity necessary to scale-down depletion methods with minimal sacrifice to proteome coverage, which makes smaller volume depletion columns desirable for maximizing sample recovery when samples are limited, as well as for reducing the expense of large-scale studies. We characterized the performance of a 346 μL column volume microscale depletion system, using four different flow rates to determine the most effective depletion conditions for ~6-μL injections of human plasma proteins and then evaluated depletion reproducibility at the optimum flow rate condition. Depletion of plasma using a commercial 10-mL depletion column served as the control. Results showed depletion efficiency of the microscale column increased as flow rate decreased, and that our microdepletion was reproducible. In an initial application, a 600-μL sample of human cerebrospinal fluid (CSF) pooled from multiple sclerosis patients was depleted and then analyzed using reversed phase liquid chromatography-mass spectrometry to demonstrate the utility of the system for this important biofluid where sample quantities are more commonly limited. PMID:25192788

  16. Detection in chick embryo of fetoproteins not recognized by the dam's immune system and of soluble alloantigens. Presumptive teratogenic and abortogenic capacity of their specific IgY

    PubMed Central

    Rodríguez-Burgos, Antonio

    2003-01-01

    Background The aim of this work was to detect antigens, non-self to the dam, potentially present in chick embryo prior to organogenesis with a view to establishing the consequences of their neutralization on chick development. To this end, hens were immunized with the extract from embryos incubated for 53 h. Their eggs were either used to isolate immunoglobulins for dot and blot tests or incubated for variable lengths of time. Results Immunoblot tests, using adsorbed primary and secondary antibodies against paternal serum, revealed the presence of at least four antigens of 32, 34, 70 and 200 kDa that can be classified as soluble alloantigens. The same antibodies against chick embryo extracts (between 53 h and 9) showed at least five aged antigens of 34, 52, 90, 200 and 250 kDa, not detected in cock serum, that can thus be considered as soluble, foreign to the immunized hens and transitory antigens. The abnormalities observed included arrested development and fetal death, as well as minor functional damage in the few chicks that were born alive. The ratio of abnormal to normal embryos was 2.85 in the experimental group and 0.43 in the control group. With regard to congenital anomalies it must be said that of the 81 eggs incubated only four chicks were born alive, and of these, only one had a healthy birth and subsequent growth. The other three showed a transitory ataxia and one of them presented adult lumbar scoliosis and asymmetric pelvis. Conclusions The problem of recurrent spontaneous abortions is revisited in the light of these results. Some recent data suggest that soluble alloantigens may be candidates for a new etiological entity in recurrent spontaneous abortions. They can also be the cause of some congenital anomalies. The soluble, foreign, transitory antigens may have a similar effect although there is no supportive data in the literature. PMID:12831405

  17. Terrestrial radioisotopes in black shale hosted Mn-carbonate deposit (Úrkút, Hungary)

    NASA Astrophysics Data System (ADS)

    Vigh, Tamás; Kovács, Tibor; Somlai, János; Kávási, Norbert; Polgári, Márta; Bíró, Lóránt

    2013-08-01

    Previously, little attention has been paid to terrestrial radioisotopes (U, Th, 40K) occurring in manganese ores, despite the fact that the biogeochemical relationship between Mn and U is versatile. Occurrence of terrestrial radioisotopes in great amounts during mining on a long-term causes significant radiation exposure. It is important to inspect black shale-hosted manganese ores from this aspect, as black shales are typically potential U-rich formations. Despite the increased radon concentration in the mine, based on the detailed major elements, trace elements and gamma spectroscopy inspection of the rock types of deposit, the U, Th enrichment was undetectable. However, the U and Th content of about average terrestrial abundance of the great ore amount may be in the background of the increased radon concentration level. This Mn-carbonate ore deposit in spite of the low U content exhibit potential radon danger for miners, which can be eliminated with intensive air change only.

  18. Reading Mustafa Kemal Atatürk on the Armenian genocide of 1915.

    PubMed

    Ulgen, Fatma

    2010-01-01

    The debate on where Mustafa Kemal Atatrk, the founder of modern Turkey and universally known as the "Father of the Turks," stood in regard to the colossal violence committed against Armenians during the First World War has become a fiercely contested part of the Turkish-Armenian reconciliation process, especially within the past few years. Ulgen aims to clear away the clouds of dust surrounding Kemal by delving into his texts and examining his role in the reification of Turkish denial of the destruction of Ottoman Armenians. Based on a textual analysis of his entire corpus, including Nutuk-the Great Speech of 1927 and the master-narrative of modern Turkish history and national identity-her article examines and documents how his charismatic leadership helped to consolidate both the myth of "murderous Armenians" and that of the Turks as an "oppressed nation" (mazlum millet), monumentalizing both in official Turkish historiography. Ulgen argues that Kemal's portrayal of Armenians and the Armenian Question was generally consistent across the years and in various political documents, as well as being consistent with contemporary Turkish representations of the events of 1915. What really tips the balance towards Turkish innocence in Kemal's representation of the conflict is not his framing of the issue per se but the stark difference in the rhetoric he deploys in depicting Armenian and Turkish atrocities and, hence, Armenians and Turks. The undeniable authority of this discursive regime is central to the resilience of Turkish denial today. PMID:20857578

  19. The growth-defense pivot: Crisis management in plants mediated by LRR-RK surface receptors

    PubMed Central

    Belkhadir, Youssef; Yang, Li; Hetzel, Jonathan; Dangl, Jeffery L.; Chory, Joanne

    2014-01-01

    Plants must adapt to their environment and require mechanisms for sensing their surroundings and responding appropriately. An expanded family of greater than 200 leucine-rich repeat receptor kinases (LRR-RKs) transduces fluctuating and often contradictory signals from the environment into changes in nuclear gene expression. Two LRR-RKs, BRASSINOSTEROID INSENSITIVE 1 (BRI1), a steroid receptor, and FLAGELLIN-SENSITIVE 2 (FLS2), an innate immune receptor that recognizes bacterial flagellin, act cooperatively to partition necessary growth-defense tradeoffs. BRI1 and FLS2 share common signaling components and slightly different activation mechanisms. BRI1 and FLS2 are paradigms for understanding signaling mechanisms of LRR-containing receptors in plants. PMID:25089011

  20. FLIPPER: a FLexIble PiPeline framEwoRk

    NASA Astrophysics Data System (ADS)

    Pierfederici, F.

    2005-12-01

    FLIPPER is a lightweight pipeline framework able to handle blackboard-based pipelines. Its ease of configuration and operation and the fact the it is extremely lightweight make FLIPPER the ideal solution for both small and large pipeline systems. FLIPPER is currently being used for the NOAO Mosaic Pipeline and has been chosen as a pipeline framework for the NOAO NEWFIRM pipeline system. The present paper describes the architecture, features, and design choices of FLIPPER in detail.

  1. Aerobic mineralization of 2,6-dichlorophenol by Ralstonia sp. strain RK1

    SciTech Connect

    Steinle, P.; Stucki, G.; Stettler, R.; Hanselmann, K.W.

    1998-07-01

    A new aerobic bacterium was isolated from the sediment of a freshwater pond close to a contaminated site at Amponville (France). It was enriched in a fixed-bed reactor fed with 2,6-dichlorophenol (2,6-DCP) as the sole carbon and energy source at pH 7.5 and room temperature. The degradation of 2,6-DCP followed Monod kinetics at low initial concentrations. At concentrations above 300 {micro}M, 2,6-DCP increasingly inhibited its own degradation. The base sequence of the 16S ribosomal DNA allowed us to assign the bacterium to the genus Ralstonia (formerly Alcaligenes). The substrate spectrum of the bacterium includes toluene, benzene, chlorobenzene, phenol, and all four ortho- and para-substituted mono- and dichlorophenol isomers. Substituents other than chlorine prevented degradation. The capacity to degrade 2,6-DCP was examined in two fixed-bed reactors. The microbial population grew on and completely mineralized 2,6-DCP at 2,6-DCP concentrations up to 740 {micro}M in continuous reactor culture supplied with H{sub 2}O{sub 2} as an oxygen source. Lack of peroxide completely stopped further degradation of 2,6-DCP. Lowering the acid-neutralizing capacity of the medium to 1/10th the original capacity led to a decrease in the pH of the effluent from 7 to 6 and to a significant reduction in the degradation activity. A second fixed-bed reactor successfully removed low chlorophenol concentrations with hydraulic residence times of 8 to 30 min.

  2. A prototype of RK/200 quantum Hall array resistance standard on epitaxial graphene

    NASA Astrophysics Data System (ADS)

    Lartsev, A.; Lara-Avila, S.; Danilov, A.; Kubatkin, S.; Tzalenchuk, A.; Yakimova, R.

    2015-07-01

    Epitaxial graphene on silicon carbide is a promising material for the next generation of quantum Hall resistance standards. Single Hall bars made of graphene have already surpassed their state-of-the-art GaAs based counterparts as an R K / 2 ( R K = h / e 2 ) standard, showing at least the same precision and higher breakdown current density. Compared to single devices, quantum Hall arrays using parallel or series connection of multiple Hall bars can offer resistance values spanning several orders of magnitude and (in case of parallel connection) significantly larger measurement currents, but impose strict requirements on uniformity of the material. To evaluate the quality of the available material, we have fabricated arrays of 100 Hall bars connected in parallel on epitaxial graphene. One out of four devices has shown quantized resistance that matched the correct value of R K / 200 within the measurement precision of 10 - 4 at magnetic fields between 7 and 9 T. The defective behaviour of other arrays is attributed mainly to non-uniform doping. This result confirms the acceptable quality of epitaxial graphene, pointing towards the feasibility of well above 90% yield of working Hall bars.

  3. Minimal Leptoquark Explanation for the R_{D^{(*)}}, R_{K}, and (g-2)_{μ} Anomalies.

    PubMed

    Bauer, Martin; Neubert, Matthias

    2016-04-01

    We show that by adding a single new scalar particle to the standard model, a TeV-scale leptoquark with the quantum numbers of a right-handed down quark, one can explain in a natural way three of the most striking anomalies of particle physics: the violation of lepton universality in B[over ¯]→K[over ¯]ℓ^{+}ℓ^{-} decays, the enhanced B[over ¯]→D^{(*)}τν[over ¯] decay rates, and the anomalous magnetic moment of the muon. Constraints from other precision measurements in the flavor sector can be satisfied without fine-tuning. Our model predicts enhanced B[over ¯]→K[over ¯]^{(*)}νν[over ¯] decay rates and a new-physics contribution to B_{s}-B[over ¯]_{s} mixing close to the current central fit value. PMID:27104699

  4. Minimal Leptoquark Explanation for the RD(*), RK, and (g -2 )μ Anomalies

    NASA Astrophysics Data System (ADS)

    Bauer, Martin; Neubert, Matthias

    2016-04-01

    We show that by adding a single new scalar particle to the standard model, a TeV-scale leptoquark with the quantum numbers of a right-handed down quark, one can explain in a natural way three of the most striking anomalies of particle physics: the violation of lepton universality in B ¯ →K ¯ ℓ+ℓ- decays, the enhanced B ¯→D(*)τ ν ¯ decay rates, and the anomalous magnetic moment of the muon. Constraints from other precision measurements in the flavor sector can be satisfied without fine-tuning. Our model predicts enhanced B ¯→K¯(*)ν ν ¯ decay rates and a new-physics contribution to Bs-B¯s mixing close to the current central fit value.

  5. A transfectant RK13 cell line permissive to classical caprine scrapie prion propagation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Classical scrapie is a form of transmissible spongiform encephalopathies (TSE) affecting domestic goats and sheep and disease is characterized by the accumulation of abnormal conformational isoform (PrP-Sc) of normal cellular prion protein (PrP-C) in the central nervous system and, in most cases, ly...

  6. Enhanced cortical thickness measurements for rodent brains via Lagrangian-based RK4 streamline computation

    NASA Astrophysics Data System (ADS)

    Lee, Joohwi; Kim, Sun Hyung; Oguz, Ipek; Styner, Martin

    2016-03-01

    The cortical thickness of the mammalian brain is an important morphological characteristic that can be used to investigate and observe the brain's developmental changes that might be caused by biologically toxic substances such as ethanol or cocaine. Although various cortical thickness analysis methods have been proposed that are applicable for human brain and have developed into well-validated open-source software packages, cortical thickness analysis methods for rodent brains have not yet become as robust and accurate as those designed for human brains. Based on a previously proposed cortical thickness measurement pipeline for rodent brain analysis,1 we present an enhanced cortical thickness pipeline in terms of accuracy and anatomical consistency. First, we propose a Lagrangian-based computational approach in the thickness measurement step in order to minimize local truncation error using the fourth-order Runge-Kutta method. Second, by constructing a line object for each streamline of the thickness measurement, we can visualize the way the thickness is measured and achieve sub-voxel accuracy by performing geometric post-processing. Last, with emphasis on the importance of an anatomically consistent partial differential equation (PDE) boundary map, we propose an automatic PDE boundary map generation algorithm that is specific to rodent brain anatomy, which does not require manual labeling. The results show that the proposed cortical thickness pipeline can produce statistically significant regions that are not observed in the previous cortical thickness analysis pipeline.

  7. Low-LET Bystander Responses in Gap Junction Null Human Colon Carcinoma Cells (RK036)

    SciTech Connect

    Sowa, Marianne B.; Goetz, Wilfried; Baulch, Janet E.; Morgan, William F.

    2006-10-01

    Combined with sensitive molecular techniques, the microbeam has proven extremely useful in studying radiation biology. This technology allows individual cells, or selected groups of cells, to be irradiated at specific doses and dose rates. Such capabilities lend themselves well to studies of low dose responses, delayed and non-targeted bystander effects of radiation. High-linear energy transfer (LET) microbeams mimic heavy energetic particles such as the ?-particles associated with radon exposure and have been used in numerous biological studies. Several low-LET microbeams have also been developed. One such low-LET microbeam, developed at the Pacific Northwest National Laboratory and currently located at the University of Maryland Radiation Oncology Research Laboratory, uses a variable energy electron source to mimic low-LET radiation and allows observation of how x-rays and ?-rays affect mammalian cells.

  8. Enhanced Cortical Thickness Measurements for Rodent Brains via Lagrangian-based RK4 Streamline Computation

    PubMed Central

    Lee, Joohwi; Kim, Sun Hyung; Oguz, Ipek; Styner, Martin

    2016-01-01

    The cortical thickness of the mammalian brain is an important morphological characteristic that can be used to investigate and observe the brain’s developmental changes that might be caused by biologically toxic substances such as ethanol or cocaine. Although various cortical thickness analysis methods have been proposed that are applicable for human brain and have developed into well-validated open-source software packages, cortical thickness analysis methods for rodent brains have not yet become as robust and accurate as those designed for human brains. Based on a previously proposed cortical thickness measurement pipeline for rodent brain analysis,1 we present an enhanced cortical thickness pipeline in terms of accuracy and anatomical consistency. First, we propose a Lagrangian-based computational approach in the thickness measurement step in order to minimize local truncation error using the fourth-order Runge-Kutta method. Second, by constructing a line object for each streamline of the thickness measurement, we can visualize the way the thickness is measured and achieve sub-voxel accuracy by performing geometric post-processing. Last, with emphasis on the importance of an anatomically consistent partial differential equation (PDE) boundary map, we propose an automatic PDE boundary map generation algorithm that is specific to rodent brain anatomy, which does not require manual labeling. The results show that the proposed cortical thickness pipeline can produce statistically significant regions that are not observed in the the previous cortical thickness analysis pipeline. PMID:27065047

  9. Characterization of Mutants Deficient in the l,d-Carboxypeptidase (DacB) and WalRK (VicRK) Regulon, Involved in Peptidoglycan Maturation of Streptococcus pneumoniae Serotype 2 Strain D39▿†

    PubMed Central

    Barendt, Skye M.; Sham, Lok-To; Winkler, Malcolm E.

    2011-01-01

    Peptidoglycan (PG) hydrolases play critical roles in the remodeling of bacterial cell walls during division. PG hydrolases have been studied extensively in several bacillus species, such as Escherichia coli and Bacillus subtilis, but remain relatively uncharacterized in ovococcus species, such as Streptococcus pneumoniae (pneumococcus). In this work, we identified genes that encode proteins with putative PG hydrolytic domains in the genome of S. pneumoniae strain D39. Knockout mutations in these genes were constructed, and the resulting mutants were characterized in comparison with the parent strain for growth, cell morphology, PG peptide incorporation, and in some cases, PG peptide composition. In addition, we characterized deletion mutations in nonessential genes of unknown function in the WalRKSpn two-component system regulon, which also contains the essential pcsB cell division gene. Several mutants did not show overt phenotypes, which is perhaps indicative of redundancy. In contrast, two new mutants showed distinct defects in PG biosynthesis. One mutation was in a gene designated dacB (spd_0549), which we showed encodes an l,d-carboxypeptidase involved in PG maturation. Notably, dacB mutants, similar to dacA (d,d-carboxypeptidase) mutants, exhibited defects in cell shape and septation, consistent with the idea that the availability of PG peptide precursors is important for proper PG biosynthesis. Epistasis analysis indicated that DacA functions before DacB in d-Ala removal, and immunofluorescence microscopy showed that DacA and DacB are located over the entire surface of pneumococcal cells. The other mutation was in WalRKSpn regulon gene spd_0703, which encodes a putative membrane protein that may function as a type of conserved streptococcal shape, elongation, division, and sporulation (SEDS) protein. PMID:21378199

  10. A NEW ALLELE, RK(CD) WITH CONVERTIBLE EXPRESSION AT THE RED KIDNEY LOCUS FOR SEEDCOAT COLOR IN COMMON BEAN

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Among light red and dark red kidney common bean (Phaseolus vulgaris L.) cultivars, pink seedcoat color (light red kidney) is dominant to dark red, but when small red cultivars (with dark red seedcoats) are crossed with dark red kidney cultivars, dark red seedcoat is dominant to the pink segregants o...

  11. Assessment of Björk-Shiley and Porcine Mitral Valve Dysfunction by M-Mode Echocardiographic Left Ventricular Dimension

    PubMed Central

    Eguaras, Manuel G.; Luque, Isabel M.; Pan, Manuel; Montero, Anastasio; Moriones, Ignacio; Granados, Jorge; Garcia, Miguel A.; Concha, Manuel

    1987-01-01

    In this study of 46 patients with mitral valve prostheses, we report on the accuracy of the left ventricular maximum filling velocity (LVMFV) index as a diagnostic tool for valve dysfunction. Echocardiographic left ventricular internal dimensions were measured every 0.05 seconds, and every measure was transferred to an axis system. The tangent to this curve was traced at the point of maximum rate of increase of dimension, and the slope in mm/sec represents the LVMFV index. Six patients, each with an obstructed mitral valve prosthesis, had a reduced LVMFV index (p < 0.001). On the other hand, eight patients with a paravalvular leak showed a LVMFV index significantly higher (p < 0.05) than patients with a normally functioning mitral valve prosthesis. The results of this study suggest that analysis of the LVMFV index can be a useful and reliable substitute for other noninvasive diagnostic techniques in evaluating certain patients with suspected prosthetic mitral valve malfunction. (Texas Heart Institute Journal 1987; 14:72-76). Images PMID:15227333

  12. Immune functions of immunoglobulin Y isolated from egg yolk of hens immunized with various infectious bacteria.

    PubMed

    Sugita-Konishi, Y; Shibata, K; Yun, S S; Hara-Kudo, Y; Yamaguchi, K; Kumagai, S

    1996-05-01

    We studied the immune functions of IgY obtained from hens immunized with a mixture of formalin-treated pathogenic bacteria. The IgY inhibited the growth of Pseudomonas aeruginosa, the production of Staphylococcus aureus enterotoxin-A, and adhesion of Salmonella enteritidis to cultured human intestinal cells (Caco 2). The results indicated that IgY specific for plural bacteria has effects useful toward prevention of bacterial diseases. PMID:8704318

  13. FEM simulations of a multi stage forming process on Sandvik maraging steel 1RK91 describing the stress assisted and the strain induced martensite transformation

    NASA Astrophysics Data System (ADS)

    Post, J.; Huétink, J.; Geijselaers, H. J. M.; Voncken, R. M. J.

    2003-10-01

    Sandvik steel IRK91 combines good corrosion resistance with high strength. The steel has good deformability in austenitic conditions. This material belongs to the group of metastable austenites, so during deformation a strain-induced transformation into martensite takes place. After deformation, transformation ccontinues as a resuit of internai stresses. Depending on the heat treatment, this stress-assisted transformation is more or less atitocatalytic. Both transformations are stress-state and temperature dependent. This article presents a constitutive model for this steel, based on the macroscopic material behaviour measured by inductive measurements. Both the stress-assisted and the strain-induced transformation to martensite are incorpomted in this model. Path-dependent work hardening is also taken into account. The model is implemented in the commercial FEM code MARC for doing simulations. In the simulations thé tools are treated as rigid bodies, friction is taken into account beeause it inflnences the stress state during metal forming. The material properties after a calculation step are mapped to the next step to incorporate the cumulative effect of the transformation and work hardening during the different steps. A multi-stage metal-forming process is simulated. The process consists of different forming steps with intervals between them to simulate the waiting time between the different metal-forming steps. Results of the transformation behaviour are presented together with the shape of the product during and after metal forming. Finally, this article shows the results of the calculation in which the material transforms autocatalytic, as a resuit of a specific heat treatment.

  14. Prospects for a soft x-ray FEL powered by a relativistic-klystron high-gradient accelerator (RK-HGA)

    SciTech Connect

    Shay, H.D.; Barletta, W.A.; Yu, S.S.; Schlueter, R.; Deis, G.A.

    1989-09-28

    We present here the concept of x-ray FELs using high gain, single-pass amplifiers with electron beams accelerated in high gradient structures powered by relativistic klystrons. Other authors have also considered x-ray FELs; the unique aspect of this paper is the use of high gradient acceleration. One of the authors has previously presented preliminary studies on this concept. The intent in this paper is to display the results of a top level design study on a high gain FEL, to present its sensitivity to a variety of fabrication and tuning errors, to discuss several mechanisms for increasing gain yet more, and to present explicitly the output characteristics of such an FEL. The philosophy of the design study is to find a plausible operating point which employs existing or nearly existing state-of-the-art technologies while minimizing the accelerator and wiggler lengths. The notion is to distribute the technical risk as evenly as possible over the several technologies so that each must advance only slightly in order to make this design feasible. This study entailed no systematic investigation of possible costs so that, for example, the sole criterion for balancing the trade-off between beam energy and wiggler length is that the two components have comparable lengths. 20 refs., 10 figs., 1 tab.

  15. TaFROG Encodes a Pooideae Orphan Protein That Interacts with SnRK1 and Enhances Resistance to the Mycotoxigenic Fungus Fusarium graminearum.

    PubMed

    Perochon, Alexandre; Jianguang, Jia; Kahla, Amal; Arunachalam, Chanemougasoundharam; Scofield, Steven R; Bowden, Sarah; Wallington, Emma; Doohan, Fiona M

    2015-12-01

    All genomes encode taxonomically restricted orphan genes, and the vast majority are of unknown function. There is growing evidence that such genes play an important role in the environmental adaptation of taxa. We report the functional characterization of an orphan gene (Triticum aestivum Fusarium Resistance Orphan Gene [TaFROG]) as a component of resistance to the globally important wheat (T. aestivum) disease, Fusarium head blight. TaFROG is taxonomically restricted to the grass subfamily Pooideae. Gene expression studies showed that it is a component of the early wheat response to the mycotoxin deoxynivalenol (DON), which is a virulence factor produced by the causal fungal agent of Fusarium head blight, Fusarium graminearum. The temporal induction of TaFROG by F. graminearum in wheat spikelets correlated with the activation of the defense Triticum aestivum Pathogenesis-Related-1 (TaPR1) gene. But unlike TaPR1, TaFROG induction by F. graminearum was toxin dependent, as determined via comparative analysis of the effects of wild-type fungus and a DON minus mutant derivative. Using virus-induced gene silencing and overexpressing transgenic wheat lines, we present evidence that TaFROG contributes to host resistance to both DON and F. graminearum. TaFROG is an intrinsically disordered protein, and it localized to the nucleus. A wheat alpha subunit of the Sucrose Non-Fermenting1-Related Kinase1 was identified as a TaFROG-interacting protein based on a yeast two-hybrid study. In planta bimolecular fluorescence complementation assays confirmed the interaction. Thus, we conclude that TaFROG encodes a new Sucrose Non-Fermenting1-Related Kinase1-interacting protein and enhances biotic stress resistance. PMID:26508775

  16. TaFROG Encodes a Pooideae Orphan Protein That Interacts with SnRK1 and Enhances Resistance to the Mycotoxigenic Fungus Fusarium graminearum1[OPEN

    PubMed Central

    Jianguang, Jia; Kahla, Amal; Arunachalam, Chanemougasoundharam; Scofield, Steven R.; Doohan, Fiona M.

    2015-01-01

    All genomes encode taxonomically restricted orphan genes, and the vast majority are of unknown function. There is growing evidence that such genes play an important role in the environmental adaptation of taxa. We report the functional characterization of an orphan gene (Triticum aestivum Fusarium Resistance Orphan Gene [TaFROG]) as a component of resistance to the globally important wheat (T. aestivum) disease, Fusarium head blight. TaFROG is taxonomically restricted to the grass subfamily Pooideae. Gene expression studies showed that it is a component of the early wheat response to the mycotoxin deoxynivalenol (DON), which is a virulence factor produced by the causal fungal agent of Fusarium head blight, Fusarium graminearum. The temporal induction of TaFROG by F. graminearum in wheat spikelets correlated with the activation of the defense Triticum aestivum Pathogenesis-Related-1 (TaPR1) gene. But unlike TaPR1, TaFROG induction by F. graminearum was toxin dependent, as determined via comparative analysis of the effects of wild-type fungus and a DON minus mutant derivative. Using virus-induced gene silencing and overexpressing transgenic wheat lines, we present evidence that TaFROG contributes to host resistance to both DON and F. graminearum. TaFROG is an intrinsically disordered protein, and it localized to the nucleus. A wheat alpha subunit of the Sucrose Non-Fermenting1-Related Kinase1 was identified as a TaFROG-interacting protein based on a yeast two-hybrid study. In planta bimolecular fluorescence complementation assays confirmed the interaction. Thus, we conclude that TaFROG encodes a new Sucrose Non-Fermenting1-Related Kinase1-interacting protein and enhances biotic stress resistance. PMID:26508775

  17. Serological evidence of Leishmania donovani infection in apparently healthy dogs using direct agglutination test (DAT) and rk39 dipstick tests in Kafta Humera, north-west Ethiopia.

    PubMed

    Kalayou, S; Tadelle, H; Bsrat, A; Abebe, N; Haileselassie, M; Schallig, H D F H

    2011-06-01

    Leishmania (Kinetoplastida: Trypanosomatidae) are protozoan parasites of significant medical and veterinary importance. Over the last decade, visceral leishmaniasis (VL) has emerged as a major opportunistic infection associated with HIV/AIDS in North Western Ethiopia. This paper reports on serological evidence of possible Leishmania donovani (L. donovani) infection in dogs using two serological tests: direct agglutination test (DAT) and Kalazar detect rapid test (KDRT). Two hundred and seventeen asymptomatic local breed dogs were examined for L. donovani antibodies. Performance of the DAT and KDRT was assessed in 162 matching samples of blood collected on filter paper and serum, respectively. Using DAT and KDRT testing in parallel, the overall seroprevalence of L. donovani infection was 27.7% and 14.8%, respectively. The degree of agreement was found to be fair (68.8%, k = 0.234). Univariable logistic regression analysis of some risk factors for L. donovani infection in dogs using DAT indicates that place of residence, sex, age, dog keeping purpose and dog housing condition were not significantly associated with seropositivity. The high proportion of positive dogs suggests the exposure of these animals to L. donovani infection and needs further investigation. Isolation and typing of the parasite aiming at confirming the role of these animals in maintenance and transmission of kala-azar is advocated. PMID:21371289

  18. TaFROG, a novel plant orphan gene, interacts with SnRK1 and enhances resistance to a mycotoxigenic fungus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    All genomes encode taxonomically restricted ‘orphan’ genes, most of which are of unknown function. We report the functional characterization of the orphan gene TaFROG as a component of the wheat resistance to the globally important Fusarium head blight (FHB) disease. TaFROG is taxonomically restrict...

  19. The influence of antismoking television advertisements on cessation by race/ethnicity, socioeconomic status, and mental health status.

    PubMed

    Nonnemaker, James M; Allen, Jane A; Davis, Kevin C; Kamyab, Kian; Duke, Jennifer C; Farrelly, Matthew C

    2014-01-01

    Disparities in tobacco use and smoking cessation by race/ethnicity, education, income, and mental health status remain despite recent successes in reducing tobacco use. It is unclear to what extent media campaigns promote cessation within these population groups. This study aims to (1) assess whether exposure to antitobacco advertising is associated with making a quit attempt within a number of population subgroups, and (2) determine whether advertisement type differentialy affects cessation behavior across subgroups. We used data from the New York Adult Tobacco Survey (NY-ATS), a cross-sectional, random-digit-dial telephone survey of adults aged 18 or older in New York State conducted quarterly from 2003 through 2011 (N = 53,706). The sample for this study consists of 9,408 current smokers from the total NY-ATS sample. Regression methods were used to examine the effect of New York State's antismoking advertising, overall and by advertisement type (graphic and/or emotional), on making a quit attempt in the past 12 months. Exposure to antismoking advertising was measured in two ways: gross rating points (a measure of potential exposure) and self-reported confirmed recall of advertisements. This study yields three important findings. First, antismoking advertising promotes quit attempts among racial/ethnic minority smokers and smokers of lower education and income. Second, advertising effectiveness is attributable in part to advertisements with strong graphic imagery or negative emotion. Third, smokers with poor mental health do not appear to benefit from exposure to antismoking advertising of any type. This study contributes to the evidence about how cessation media campaigns can be used most effectively to increase quit attempts within vulnerable subgroups. In particular, it suggests that a general campaign can promote cessation among a range of sociodemographic groups. More research is needed to understand what message strategies might work for those with poor

  20. NASA's Origins and the Dawn of the Space Age. No. 10; Monographs in Aerospace History

    NASA Technical Reports Server (NTRS)

    Portree, David S. F.

    1998-01-01

    The twenty page narrative describes historical circumstances around Sputnik, the International Geophysical Year (IGY) and the formation of NASA from NACA in 1957-1958. Appendices include reproductions of relevant historical documents.

  1. Using immunoglobulin Y as an alternative antibody for the detection of hepatitis A virus in frozen liver sections.

    PubMed

    Bentes, Gentil Arthur; Lanzarini, Natália Maria; Lima, Lyana Rodrigues Pinto; Manso, Pedro Paulo de Abreu; da Silva, Alexandre Dos Santos; Mouta Junior, Sergio da Silva E; Guimarães, Juliana Rodrigues; de Moraes, Marcia Terezinha Baroni; Pelajo-Machado, Marcelo; Pinto, Marcelo Alves

    2015-06-01

    An increasing amount of research has been conducted on immunoglobulin Y (IgY) because the use of IgY offers several advantages with respect to diagnostic testing, including its easy accessibility, low cost and translatability to large-scale production, in addition to the fact that it can be ethically produced. In a previous work, immunoglobulin was produced and purified from egg yolks (IgY) reactive to hepatitis A virus (HAV) antigens. In the present work, this anti-HAV-specific IgY was used in an indirect immunofluorescence assay to detect viral antigens in liver biopsies that were obtained from experimentally infected cynomolgus monkeys. Fields that were positive for HAV antigen were detected in liver sections using confocal microscopy. In conclusion, egg yolks from immunised hens may be a reliable source for antibody production, which can be employed for immunological studies. PMID:25993400

  2. Using immunoglobulin Y as an alternative antibody for the detection of hepatitis A virus in frozen liver sections

    PubMed Central

    Bentes, Gentil Arthur; Lanzarini, Natália Maria; Lima, Lyana Rodrigues Pinto; Manso, Pedro Paulo de Abreu; da Silva, Alexandre dos Santos; Mouta, Sergio da Silva e; Guimarães, Juliana Rodrigues; de Moraes, Marcia Terezinha Baroni; Pelajo-Machado, Marcelo; Pinto, Marcelo Alves

    2015-01-01

    An increasing amount of research has been conducted on immunoglobulin Y (IgY) because the use of IgY offers several advantages with respect to diagnostic testing, including its easy accessibility, low cost and translatability to large-scale production, in addition to the fact that it can be ethically produced. In a previous work, immunoglobulin was produced and purified from egg yolks (IgY) reactive to hepatitis A virus (HAV) antigens. In the present work, this anti-HAV-specific IgY was used in an indirect immunofluorescence assay to detect viral antigens in liver biopsies that were obtained from experimentally infected cynomolgus monkeys. Fields that were positive for HAV antigen were detected in liver sections using confocal microscopy. In conclusion, egg yolks from immunised hens may be a reliable source for antibody production, which can be employed for immunological studies. PMID:25993400

  3. Passive immune-protection of small abalone against Vibrio alginolyticus infection by anti-Vibrio IgY-encapsulated feed.

    PubMed

    Wu, Chang-Jer; Wang, Hang; Chan, Yi-Lin; Li, Tsung-Lin

    2011-01-01

    Small abalone (Haliotis diversicolor supertexta) is a high value-added shellfish. It however has been suffering Vibrio alginolyticus infections, which cause mass death of small abalone and thus great economic losses, particularly in artificial aquaculture. In this study, we attempted to treat small abalone with anti-Vibrio IgY to elicit a passive immunity directly against V. alginolyticus infections. Anti-Vibrio IgY was alginate encapsulated in egg powders as feed, which may avoid antibody inactivation in the gastrointestinal tract of small abalone. The feed was tested for the stability of anti-Vibrio IgY in a gastrointestinal mimic environment. The result showed anti-Vibrio IgY retained activity as high as 90% after 4 h exposure to pancreatic enzymes. Addition of 0, 5 or 10% anti-Vibrio IgY-encapsulated egg powders into a basal diet to form abalone diet formulae. Small abalones fed with the anti-Vibrio IgY formulae showed a relatively high respiratory burst activity than those without anti-Vibrio IgY treatments. The survival rates of small abalones fed with 5 or 10% anti-Vibrio IgY egg powders were in the range of 65-70% 14 days post-V. alginolyticus challenge (1 x 10⁶ c.f.u.), which was significantly higher than 0% of those fed without anti-Vibrio IgY. The anti-Vibrio IgY-encapsulated formulae were thus concluded to be an effective means to prevent small abalone from V. alginolyticus infection, and may be practical in use in abalone aquaculture. PMID:21300158

  4. Corneal topography analysis before and after radial keratotomy.

    PubMed

    Lin, Y; Chen, J; Wang, Z; Xuan, J

    1995-05-01

    Corneal topographic analyses were conducted on 58 eyes of 42 cases with mild or moderate myopia by computer-assisted photokeratography before and after radial keratotomy (RK). The results indicate that the corneal surfaces of most examined eyes (76%) are positive aspheric shape before RK, while after RK they are changed to negative aspheric shape. Before RK, the corneal topography of most examined eyes (67%) is symmetric bow tie pattern or asymmetric bow tie pattern; after RK, the forms of central flattened regions are mainly round or approximately round, dumbbell and belt-shaped. Before and after RK there is no significant change in surface regularity index (SRI), but surface asymmetry index (SAI) is changed significantly. RK makes the central and central peripheral parts of the cornea flattened. The most significant RK effect is found at the region 1.140 +/- 0.090 mm away from the corneal optical center. Examination and analysis on the corneal topography before and after RK not only provide an accurate and objective basis for RK operational plan, but also make it possible to objectively and quantitatively evaluate the RK effect, to accurately predict the RK clinical effects, and to effectively improve the RK clinical quality. PMID:7555399

  5. Green turtles (Chelonia mydas) have novel asymmetrical antibodies

    USGS Publications Warehouse

    Work, Thierry M.; Dagenais, Julie; Breeden, Renee; Schneemann, Anette; Sung, Joyce; Hew, Brian; Balazs, George H.; Berestecky, John M.

    2015-01-01

    Igs in vertebrates comprise equally sized H and L chains, with exceptions such as H chain–only Abs in camels or natural Ag receptors in sharks. In Reptilia, Igs are known as IgYs. Using immunoassays with isotype-specific mAbs, in this study we show that green turtles (Chelonia mydas) have a 5.7S 120-kDa IgY comprising two equally sized H/L chains with truncated Fc and a 7S 200-kDa IgY comprised of two differently sized H chains bound to L chains and apparently often noncovalently associated with an antigenically related 90-kDa moiety. Both the 200- and 90-kDa 7S molecules are made in response to specific Ag, although the 90-kDa molecule appears more prominent after chronic Ag stimulation. Despite no molecular evidence of a hinge, electron microscopy reveals marked flexibility of Fab arms of 7S and 5.7S IgY. Both IgY can be captured with protein G or melon gel, but less so with protein A. Thus, turtle IgY share some characteristics with mammalian IgG. However, the asymmetrical structure of some turtle Ig and the discovery of an Ig class indicative of chronic antigenic stimulation represent striking advances in our understanding of immunology.

  6. The International Geophysical Year: Its influence on the beginning of the French space program

    NASA Astrophysics Data System (ADS)

    Moulin, Hervé

    2010-03-01

    In 1957-1958, the International Geophysical Year (IGY) was the most important scientific cooperation programme in the World, after the Second World War. Thousands of scientists from 67 countries were involved in this large operation, among them a lot of French scientists. IGY was previously called the IPY (International Polar Year) and France, as many other countries, has been involved in the Arctic and Antarctic regions researches. Everybody knows that the IGY is at the origin of Sputnik and the first launch of Russian and American satellites. But, we know less about the IGY rockets programme itself in which France had intended to participate. This paper will discuss this programme with a special highlight on some aspects of the French participation and their relationship with the IGY programme. This approach arises several questions, such as: Which French scientists have been involved? What was the attitude of the French Government about this program, etc. We focus our analysis on the interrogation: did the IGY have any real influence on the origin of the French space research activities?

  7. Production, Characterization and Applications for Toxoplasma gondii-Specific Polyclonal Chicken Egg Yolk Immunoglobulins

    PubMed Central

    Ferreira Júnior, Álvaro; Santiago, Fernanda M.; Silva, Murilo V.; Ferreira, Flávia B.; Macêdo Júnior, Arlindo G.; Mota, Caroline M.; Faria, Matheus S.; Filho, Hercílio H. Silva; Silva, Deise A. O.; Cunha-Júnior, Jair P.; Mineo, José R.; Mineo, Tiago W. P.

    2012-01-01

    Background Toxoplasma gondii may cause abortions, ocular and neurological disorders in warm-blood hosts. Immunized mammals are a wide source of hyperimmune sera used in different approaches, including diagnosis and the study of host-parasite interactions. Unfortunately, mammalian antibodies present limitations for its production, such as the necessity for animal bleeding, low yield, interference with rheumatoid factor, complement activation and affinity to Fc mammalian receptors. IgY antibodies avoid those limitations; therefore they could be an alternative to be applied in T. gondii model. Methodology/Principal Findings In this study we immunized hens with soluble tachyzoite antigens of T. gondii (STAg) and purified egg yolk antibodies (IgY) by an inexpensive and simple method, with high yield and purity degree. IgY anti-STAg antibodies presented high avidity and were able to recognize a broad range of parasite antigens, although some marked differences were observed in reactivity profile between antibodies produced in immunized hens and mice. Interestingly, IgY antibodies against Neospora caninum and Eimeria spp. did not react to STAg. We also show that IgY antibodies were suitable to detect T. gondii forms in paraffin-embedded sections and culture cell monolayers. Conclusions/Significance Due to its cost-effectiveness, high production yield and varied range of possible applications, polyclonal IgY antibodies are useful tools for studies involving T. gondii. PMID:22808150

  8. Green Turtles (Chelonia mydas) Have Novel Asymmetrical Antibodies.

    PubMed

    Work, Thierry M; Dagenais, Julie; Breeden, Renee; Schneemann, Anette; Sung, Joyce; Hew, Brian; Balazs, George H; Berestecky, John M

    2015-12-01

    Igs in vertebrates comprise equally sized H and L chains, with exceptions such as H chain-only Abs in camels or natural Ag receptors in sharks. In Reptilia, Igs are known as IgYs. Using immunoassays with isotype-specific mAbs, in this study we show that green turtles (Chelonia mydas) have a 5.7S 120-kDa IgY comprising two equally sized H/L chains with truncated Fc and a 7S 200-kDa IgY comprised of two differently sized H chains bound to L chains and apparently often noncovalently associated with an antigenically related 90-kDa moiety. Both the 200- and 90-kDa 7S molecules are made in response to specific Ag, although the 90-kDa molecule appears more prominent after chronic Ag stimulation. Despite no molecular evidence of a hinge, electron microscopy reveals marked flexibility of Fab arms of 7S and 5.7S IgY. Both IgY can be captured with protein G or melon gel, but less so with protein A. Thus, turtle IgY share some characteristics with mammalian IgG. However, the asymmetrical structure of some turtle Ig and the discovery of an Ig class indicative of chronic antigenic stimulation represent striking advances in our understanding of immunology. PMID:26500346

  9. Development of microbeads of chicken yolk antibodies against Clostridium difficile toxin A for colonic-specific delivery.

    PubMed

    Zhang, Shumin; Xing, Pingping; Guo, Guiping; Liu, Hong; Lin, Donghai; Dong, Chuangchuang; Li, Min; Feng, Dongxiao

    2016-07-01

    The incidence of Clostridium difficile infection has increased in Western world in the past 10 years, similar infection rates are also reported in developing countries such as China. Current antibiotics treatments have recurrence rates between 15% and 30%. IgY antibodies against toxin A of C. difficile could protect animal models from the challenge of lethal dose of C. difficile spores. However, IgY is sensitive to the low pH environment of the stomach and proteinases in the intestine. The objective of this study was to prepare colonic-specific delivery system of toxin A antigen-specific IgY to block the recognition of toxin A to the colon mucosa cells. Egg-laying hens were immunized with purified C. difficile toxin A C-terminal domain for 3 times, then egg IgY against the recombinant ToxA-C protein was purified from immunized egg yolk and frozen dried. IgY-loaded microbeads were prepared using mini fluid bed system; the loading efficiency was 21%. The pH and temperature stabilities of the microbeads were assayed. The IgY-loaded microbeads coated with 35% Eudragit S100 had colonic-specific IgY release specificity both in vitro and in vivo, the colonic-specific release of biological active IgY was 87.5% in the rat. Our study provides a new option for the biological treatment C. difficile infection. PMID:25799315

  10. A simple method for isolating chicken egg yolk immunoglobulin using effective delipidation solution and ammonium sulfate.

    PubMed

    Tong, Chenyao; Geng, Fang; He, Zhenjiao; Cai, Zhaoxia; Ma, Meihu

    2015-01-01

    Chicken egg yolk immunoglobulin (IgY) is a superior alternative to mammalian immunoglobulin. However, the practical application of IgY in research, diagnostics, and functional food is limited due to complex or time-consuming purification procedures. The objective of this study was to develop a simple, safe, large-scale separation method for IgY from egg yolk. Egg yolk was diluted with 6-fold delipidation solutions made of different types (pectin, λ-carrageenan, carboxymethylcellulose, methylcellulose, and dextran sulfate) and concentrations (0.01, 0.05, 0.1, 0.15, and 0.2%) of polysaccharides, respectively. The yolk solution was adjusted to pH 5.0, and then kept overnight at 4°C before being centrifuged at 4°C. The resulting supernatant was added to 35% (w/v) (NH4)2SO4 and then centrifuged. The precipitant, which contained IgY, was dissolved in distilled water and then dialyzed. SDS-PAGE and Western blotting were utilized to conduct qualitative analysis of IgY; high-performance liquid chromatography (HPLC) was used for quantitative analysis. The immunoreactivity of IgY was measured by ELISA. The results showed that yield, purity, and immunoreactivity varied with types and concentrations of polysaccharides. The optimal isolation of IgY for pectin, λ-carrageenan, dextran sulfate, and carboxymethylcellulose was at the concentration of 0.1%; for methylcellulose, optimal isolation was at 0.15%. The best results were obtained in the presence of 0.1% pectin. In this condition, yield and purity can reach 8.36 mg/mL egg yolk and 83.3%, respectively, and the negative effect of IgY on immunoreactivity can be minimized. The procedure of isolation was simplified to 2 steps with a higher yield of IgY, avoiding energy- and time-consuming methods. Therefore, the isolation condition under study has a great potential for food industry production of IgY on a large scale. PMID:25542196