Mannitol transport and mannitol dehydrogenase activities are coordinated in olea europaea under salt and osmotic stresses

This is a pre-copy-editing, author-produced PDF of an article accepted for publication in Plant and Cell Physiology following peer review. The definitive publisher-authenticated version is available online at http://pcp.oxfordjournals.org/cgi/content/abstract/pcr121? ijkey=6orgUM5fkIjedYn&keytype=refThe intracellular accumulation of organic compatible solutes functioning as osmoprotectants, such as polyols, is an important response mechanism of several plants to drought and salinity. In Olea europaea a mannitol transport system (OeMaT1) was previously characterised as a key player in plant response to salinity. In the present study, heterotrophic sink models, such as olive cell suspensions and fruit tissues, and source leaves were used for analytical, biochemical and molecular studies. The kinetic parameters of mannitol dehydrogenase (MTD) determined in mannitol-growing cells, at 25 °C and pH 9.0, were as follows: Km, 54.5 mM mannitol and Vmax, 0.47 μmol h-1 mg-1 protein. The corresponding cDNA was cloned and named OeMTD1. OeMTD1 expression was correlated with MTD activity, OeMaT1 expression and carrier-mediated mannitol transport, in mannitol- and sucrose-growing cells. Furthermore, sucrosegrowing cells displayed only residual OeMTD activity, even though high levels of OeMTD1 transcription were observed. There is evidence OeMTD is regulated at both transcriptional and post-transcriptional levels. MTD activity and OeMTD1 expression were repressed after Na+, K+ and PEG treatments, both in mannitol- and sucrose-growing cells. In contrast, salt and drought significantly increased mannitol transport activity and OeMaT1 expression. Altogether, these studies support that olive tree copes with salinity and drought by coordinating mannitol transport with intracellular metabolism.This work was supported by the Portuguese Foundation for Science and Technology (FCT) (research project ref. PTDC/AGR-ALI/100636/2008; to A. Conde, grant ref. SFRH/BD/47699/2008; to C. Conde, grant ref. SFRH/BPD/34998/2007; to P. Silvagrant ref. SFRH/BD/13460/2003)

Efficacy of topical cobalt chelate CTC-96 against adenovirus in a cell culture model and against adenovirus keratoconjunctivitis in a rabbit model

BACKGROUND: Adenovirus (Ad), associated with significant morbidity, has no topical treatment. A leading CTC compound (CTC-96), a Co(III )chelate, was found to have potent in vitro and in vivo antiviral efficacy against herpes viruses. In this study CTC-96 is being tested for possible anti-Adenovirus activity. METHODS: The biological anti-adenovirus activity of CTC-96 in concentrations from 5 to 250 ug/ml, was evaluated initially by viral inactivation (viral exposure to CTC-96 followed by dilution and inoculation of cells), virucidal (viral exposure to CTC-96 and inoculation of cells without dilution) and antiviral (effect of CTC-96 on previously adsorbed virus) plaque assays on HeLa (human cervical carcinoma), A549 (human lung carcinoma) and SIRC (rabbit corneal) cells. After verifying the antiviral activity, New Zealand White rabbits were infected with Ad-5 into: 1) the anterior cul-de-sac scarifying the conjunctiva (Group "C+"); 2) the anterior cul-de-sac scarifying the conjunctiva and cornea (Group "CC+"); 3) the stroma (Group "CI+"). Controls were sham-infected ("C-", "CC-", "CI-"). Other rabbits, after "CC", were treated for 21 days with: 1) placebo, 9x/day ("-"); 2) CTC-96, 50 ug/ml, 9x/day ("50/9"); CTC-96, 50 ug/ml, 6x/day ("50/6"); CTC-96, 25 ug/ml, 6x/day ("25/6"). All animals were monitored via examination and plaque assays. RESULTS: In vitro viral inactivation, virucidal and antiviral assays all demonstrated CTC-96 to be effective against Adenvirus type 5 (ad-5). The in vivo model of Ad keratoconjunctivitis most similar to human disease and producing highest viral yield was "CC". All eyes (6/6) developed acute conjunctivitis. "CI" yielded more stromal involvement (1/6) and iritis (5/6), but lower clinical scores (area × severity). Infection via "C" was inconsistent (4/6). Fifty (50) ug/ml was effective against Ad-5 at 6x, 9x dosings while 25 ug/ml (6x) was only marginally effective. CONCLUSION: CTC-96 demonstrated virucidal activity against Ad5 in tissue culture with HeLa, A549 and SIRC cell lines. Animal Model Development: 1) "CC" produced conjunctival infection with occasional keratitis similar to human disease; "CI" yielded primarily stromal involvement; 2) "C" consistently produced neither conjunctivitis nor keratitis. CTC Testing: 1) Conjunctivitis in all eyes; 2) Resolution fastest in "50/9" ("50/9". "50/6" > "25/6" > "-"); 3) Efficacy in "50/6" was not statistically different than "50/9"; 4) Conjunctival severity was lower in treatment groups then controls; 5) Little corneal or intra-ocular changes were noted

Poly(ADP-Ribose) Polymerase 1 (PARP-1) Regulates Ribosomal Biogenesis in Drosophila Nucleoli

Poly(ADP-ribose) polymerase 1 (PARP1), a nuclear protein, utilizes NAD to synthesize poly(AD-Pribose) (pADPr), resulting in both automodification and the modification of acceptor proteins. Substantial amounts of PARP1 and pADPr (up to 50%) are localized to the nucleolus, a subnuclear organelle known as a region for ribosome biogenesis and maturation. At present, the functional significance of PARP1 protein inside the nucleolus remains unclear. Using PARP1 mutants, we investigated the function of PARP1, pADPr, and PARP1-interacting proteins in the maintenance of nucleolus structure and functions. Our analysis shows that disruption of PARP1 enzymatic activity caused nucleolar disintegration and aberrant localization of nucleolar-specific proteins. Additionally, PARP1 mutants have increased accumulation of rRNA intermediates and a decrease in ribosome levels. Together, our data suggests that PARP1 enzymatic activity is required for targeting nucleolar proteins to the proximity of precursor rRNA; hence, PARP1 controls precursor rRNA processing, post-transcriptional modification, and pre-ribosome assembly. Based on these findings, we propose a model that explains how PARP1 activity impacts nucleolar functions and, consequently, ribosomal biogenesis

I hope all my vegetarian friends forgive me

Mark_Winicov__Philadelphia__PA__I_hope_all_my_vegetarian_friends_forgive_me_February_05__2014_at_0431PMThese images were created as part of the Hughes Remix project, a collaborative endeavor developed by UMBC's Albin O. Kuhn Library Special Collections and the Department of Visual Art to foster creative engagement with archival holdings in conjunction with the 2014 Society for Photographic Education annual conference. UMBC's Special Collections offered a selection of images from the Hughes Company Glass Negatives (http://contentdm.ad.umbc.edu/cdm/landingpage/collection/hughes) collection for SPE members and conference attendees to remix, reinvent, reinterpret, and reimagine the images in this collection of Baltimore street scenes, promotional and advertising photographs, businesses, churches, schools, monuments, factories, machinery, and portraits. Images that were created were displayed during the conference as well as on a Tumblr site and are now archived in the Special Collections. Full details of the project can be found at: http://hughes-remix.tumblr.com/overview.The original Hughes Company Glass Negatives collection can be found at: http://contentdm.ad.umbc.edu/cdm/landingpage/collection/hughe