Analysis of the chlorophyll catabolism pathway in leaves of an introgression senescence mutant of Lolium temulentum

Maria Roca, Caron James, Adriana Pruzinsk?, Stefan H?rtensteiner, Howard Thomas and Helen Ougham. Analysis of the chlorophyll catabolism pathway in leaves of an introgression senescence mutant of Lolium temulentum. Phytochemistry, 65 (9), 1231-1238. Sponsorship: BBSRC RAE2008

Identification and analysis of mechanisms involved in a broad-spectrum disease resistant mutant of the winter wheat cv. Guardian

M66 an X-ray induced mutant of winter wheat (Triticum aestivum) cv. Guardian exhibits broad-spectrum resistance to powdery mildew (Blumeria graminis f. sp. tritici), yellow rust (Puccinia striiformis f. sp. tritici), and leaf rust (Puccinia recondita f. sp. tritici), along with partial resistance to stagnonospora nodorum blotch (caused by the necrotroph Stagonosporum nodorum) and septoria tritici blotch (caused by the hemibiotroph Mycosphaerella graminicola) compared to the parent plant ���Guardian���. Analysis revealed that M66 exhibited no symptoms of infection following artificial inoculation with Bgt in the glasshouse after adult growth stage (GS 45). Resistance in M66 was associated with widespread leaf flecking which developed during tillering. Flecking also occurred in M66 leaves without Bgt challenge; as a result grain yields were reduced by approximately 17% compared to ���Guardian��� in the absence of disease. At the seedling stage, M66 exhibited partial resistance. M66, along with Tht mutants (Tht 12, Tht13), also exhibit increased tolerance to environmental stresses (abiotic), such as drought and heat stress at seedling and adult growth stages, However, adult M66 exhibited increased susceptibility to the aphid Schizaphis graminum compared to ���Guardian���. Resistance to Bgt in M66 was characterized with increased and earlier H2O2 accumulation at the site of infection which resulted in increased papilla formation in epidermal cells, compared to ���Guardian���. Papilla formation was associated with reduced pathogen ingress and haustorium formation, indicating that the primary cause of resistance in M66 was prevention of pathogen penetration. Heat treatment at 46�� C prior to challenge with Bgt also induced partial disease resistance to Blumeria graminis f. sp. tritici in ���Guardian��� and M66 seedlings. This was characterized by a delay in primary infection, due to increased production of ROS species, such as hydrogen peroxide, ROS-scavenging enzymes and Hsp70, resulting in cross-linking of cell wall components prior to inoculation. This actively prevented the fungus from penetrating the epidermal cell wall. Proteomics analysis using 2-D gel electrophoresis identified primary and secondary disease resistance effects in M66 including detection of ROS scavenging enzymes (4, 24 hai), such as ascorbate peroxidase and a superoxidase dismutase isoform (CuZnSOD) in M66 which were absent from ���Guardian���. Chitinase (PR protein) was also upregulated (24 hai) in M66 compared to ���Guardian���.Monosomic and ditelosomic analysis of M66 revealed that the mutation in M66 is located on the long arm of chromosome 2B (2BL). Chromosome 2BL is known to have key genes involved in resistance to pathogens such as those causing stripe rust and powdery mildew. The TaMloB1 gene, an orthologue of the barley Mlo gene, is also located on chromosome 2BL. Sanger sequencing of part of the coding sequence revealed no deletions in the TaMloB1 gene between ���Guardian��� and M66.

Structured Illumination Microscopy and a Quantitative Image Analysis for the Detection of Positive Margins in a Pre-Clinical Genetically Engineered Mouse Model of Sarcoma.

Intraoperative assessment of surgical margins is critical to ensuring residual tumor does not remain in a patient. Previously, we developed a fluorescence structured illumination microscope (SIM) system with a single-shot field of view (FOV) of 2.1 �� 1.6 mm (3.4 mm2) and sub-cellular resolution (4.4 ��m). The goal of this study was to test the utility of this technology for the detection of residual disease in a genetically engineered mouse model of sarcoma. Primary soft tissue sarcomas were generated in the hindlimb and after the tumor was surgically removed, the relevant margin was stained with acridine orange (AO), a vital stain that brightly stains cell nuclei and fibrous tissues. The tissues were imaged with the SIM system with the primary goal of visualizing fluorescent features from tumor nuclei. Given the heterogeneity of the background tissue (presence of adipose tissue and muscle), an algorithm known as maximally stable extremal regions (MSER) was optimized and applied to the images to specifically segment nuclear features. A logistic regression model was used to classify a tissue site as positive or negative by calculating area fraction and shape of the segmented features that were present and the resulting receiver operator curve (ROC) was generated by varying the probability threshold. Based on the ROC curves, the model was able to classify tumor and normal tissue with 77% sensitivity and 81% specificity (Youden's index). For an unbiased measure of the model performance, it was applied to a separate validation dataset that resulted in 73% sensitivity and 80% specificity. When this approach was applied to representative whole margins, for a tumor probability threshold of 50%, only 1.2% of all regions from the negative margin exceeded this threshold, while over 14.8% of all regions from the positive margin exceeded this threshold.

LibQUAL+ Survey 2004 @ Queen's University Library : analysis of results

Queen's University Library was one of 202 libraries, including 57 members of the Association of Research Libraries (ARL), to survey its users in spring 2004 using the LibQUAL+ survey instrument. LibQUAL+ was designed by ARL to assist libraries in assessing the quality of their services and identifying areas for improvement. # Overall: Queen's scored higher than the average for all ARL participants and 1st among the 2004 Canadian participants. This relatively high rating is due to very high scores in the dimensions of Library as Place and Affect of Service. However, there is considerable need for improvement in the area of Information Control where Queen's rated well below the ARL average. # Affect of Service: Queen's strong overall ratings are supported by the many respondent comments praising customer service throughout the system. The ratings and survey comments indicate greatest appreciation by faculty and more experienced students (e.g. graduate students) for the instruction and on-site services provided by the libraries. The ratings also indicate that undergraduates, growing up with the web, want and expected to be able to access library resources independently and do not value these services as highly. The comments also indicated some specific areas for improvement throughout the library system. # Library as Place : All Queen's libraries except for Law ranked well above the ARL and Canadian averages. Overall, Library as Place ranked lowest in importance among the service dimensions for all ARL participants including Queen's. Comparative analysis of LibQUAL results since the survey began shows a decline in ���desired��� ratings for Library as Place. However, undergraduates continue to give strong "desired" ratings to certain aspects of Library as Place and a relatively high rating for "minimum expected" service. The comments from Queen's survey respondents and ARL's analyses of focus groups indicate that undergraduates value the library much more as a place to study and work with peers rather than for its on-site resources and services. # Information Control: This is the area in greatest need of attention. While it ranked highest in importance for all user groups by a wide margin, Queen's performed poorly in this category. Overall, Queen's ranked far below both the ARL average and the top three Canadian scores. However, the major dissatisfaction was concentrated in the humanities/social sciences (Stauffer primary users) and the health sciences (Bracken primary users) where the overall rating of perceived service quality ranked below the minimum expected service rating. Primary users of the Education, Engineering/Science and Law libraries rated this service dimension higher than the ARL average. The great success of the Canadian National Site License Program (CNSLP) is reflected in the high overall rating generated by Engineering/Science Library users. The low ratings from the humanities and social sciences are supported by respondents' comments and are generally consistent with other ARL participants.

A novel Kazal-type trypsin inhibitor from the skin secretion of the Central American red-eyed leaf frog, Agalychnis callidryas.

The chemical complexity of the defensive skin secretion of the red-eyed leaf frog, (Agalychnis callidryas), has not been elucidated in detail. During a systematic study of the skin secretion peptidomes of phyllomedusine frogs, we discovered a novel Kazal-type protein with potent trypsin inhibitory activity (Ki = 1.9 nM) that displays the highest degree of structural similarity with Kazal proteins from bony fishes. The protein was located in reverse-phase HPLC fractions following a screen of such for trypsin inhibition and subsequent partial Edman degradation of the peak active fraction derived the sequence: ATKPR-QYIVL-PRILRPV-GT. The molecular mass of the major component in this fraction was established by MALDI-TOF MS as 5893.09 Da. This partial sequence (assuming blank cycles to be Cys residues) was used to design a degenerate primer pool that was employed successfully in RACE-PCR to clone homologous precursor-encoding cDNA that encoded a mature Kazal protein of 52 amino acid residues with a computed molecular mass of 5892.82 Da. The protein was named A. callidryas Kazal trypsin inhibitor (ACKTI). BLAST analysis revealed that ACKTI contained a canonical Kazal motif (C-x(7)-C-x(6)-Y-x(3)-C-x(2,3)-C). This novel amphibian skin Kazal trypsin inhibitor adds to the spectrum of trypsin inhibitors of Kunitz- and Bowman Birk-type reported from this amphibian source.

PsT-1: A new Tryptophyllin peptide from the skin secretion of Waxy Monkey Leaf Frog, Phyllomedusa sauvagei

The Waxy Monkey Leaf Frog, Phyllomedusa sauvagei, has been extensively-studied for many years, and a broad spectrum of bioactive peptides has been found in its skin secretions. Here we report the discovery of a novel tryptophyllin (TPH) peptide, named PsT-1, from this frog species. Skin secretions from specimens of P. sauvagei were collected by mild electrical stimulation. Peptides were identified and characterized by transcriptome cloning, and the structure was confirmed by MALDI-TOF mass spectrometry and automated Edman degradation. This novel peptide was encoded by a single precursor of 61 amino acid residues, whose primary structure was deduced from cloned skin cDNA. Analysis of different amphibian tryptophyllins revealed that PsT-1 exhibited a high degree of primary structural similarity to its homologues, PdT-1 and PdT-2, from the Mexican giant leaf frog, Pachymedusa dacnicolor. A synthetic replicate of PsT-1 was found to inhibit bradykinin-induced vasorelaxation of phenylephrine pre-constricted rat tail artery smooth muscle. It was also found that PsT-1 had an anti-proliferative effect on three different human prostate cancer cell lines (LNCaP/PC3/DU145), by use of an MTT assay coupled with direct cell counting as measures of cell growth. These data indicate that PsT-1 is a likely bradykinin receptor antagonist and its biological effects are probably mediated through bradykinin receptors. As a BK antagonist, PST-1, with antagonistic effects on BK in artery smooth muscle, inhibition of proliferation in prostate cancer cells and lack of undesirable side effects, may have potential in cardiovascular, inflammatory and anticancer therapy.

Multi-scale dynamics on the Eastern Brazilian margin

Este trabalho combina esfor��os de simula����o num��rica e de an��lise de dados para investigar a din��mica em diversos compartimentos (oceano aberto, plataforma continental e zona costeira-estuarina) e, em multiplas escalas, na Margem Continental Leste Brasileira (MCLB). A circula����o de largo e mesoescala espacial e a propaga����o da mar�� barotr��pica s��o investigadas atrav��s de uma configura����o aninhada do modelo num��rico ROMS. O estudo da din��mica regional da Ba��a de Camamu (CMB) baseia-se na an��lise de dados locais. A MCLB, localizada a SW do Atl��ntico Sul entre 8��S e 20��S, possui plataforma estreita, batimetria complexa, e baixa produtividade prim��ria. A sua din��mica �� influenciada pela diverg��ncia da Corrente Sul Equatorial (CSE). As simula����es refletem as conex��es sazonais e espaciais entre a Corrente do Brasil e a Contra Corrente Norte do Brasil , em conex��o com a din��mica da CSE. As simula����es revelam atividades vorticais nas proximidades da costa e intera����es com a din��mica costeira, cujos padr��es s��o descritos. A valida����o do modelo em mesoescala �� baseada em c��lculos de energia cin��tica turbulenta e em dados hist��ricos de transporte. A CMB, localizada a 13��400S, abriga uma comunidade piscat��ria tradicional e extenso de manguezal. Situa-se por��m sobre uma bacia sedimentar com grande reservas de ��leo e g��s, estando em tens��o permanente de impacto ambiental. Neste trabalho sumarizamos as condi����es f��sicas regionais e investigamos sua din��mica interna, focando sua variabilidade em amostragens realizadas sob condi����es de seca (Setembro de 2004) e de chuva (Julho de 2005). Finalmente, o modelo num��rico ROMS �� for��ado com o sinal de mar��, empregando-se uma configura����o simples (com coeficientes de atrito de fundo constantes e condi����es hidrogr��ficas homog��neas), com o intuito de avaliar sua resposta e investigar a natureza da propaga����o da mar�� barotr��pica na MCLB, convergindo na CMB. A an��lise da resposta do modelo �� mar�� basea-se em s��ries hist��ricas do n��vel do mar para a MCLB e dados recentes da CMB.

Antioxidant properties and phenol content of Portulaca oleracea L. leaf, stems and flowers infusions: health benefits

Disserta����o de mest., Ci��ncias Biom��dicas, Departamento de Ci��ncias Biom��dicas e Medicina, Univ. do Algarve, 2011

Business model analysis of Mobitto: How to solve the chicken and egg problem of a multi-sided platform?

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Management from the NOVA ��� School of Business and Economics

Analysis of temporal and spatial variations of forest. A case of study in notheastern Armenia

The forest has a crucial ecological role and the continuous forest loss can cause colossal effects on the environment. As Armenia is one of the low forest covered countries in the world, this problem is more critical. Continuous forest disturbances mainly caused by illegal logging started from the early 1990s had a huge damage on the forest ecosystem by decreasing the forest productivity and making more areas vulnerable to erosion. Another aspect of the Armenian forest is the lack of continuous monitoring and absence of accurate estimation of the level of cuts in some years. In order to have insight about the forest and the disturbances in the long period of time we used Landsat TM/ETM + images. Google Earth Engine JavaScript API was used, which is an online tool enabling the access and analysis of a great amount of satellite imagery. To overcome the data availability problem caused by the gap in the Landsat series in 1988- 1998, extensive cloud cover in the study area and the missing scan lines, we used pixel based compositing for the temporal window of leaf on vegetation (June-late September). Subsequently, pixel based linear regression analyses were performed. Vegetation indices derived from the 10 biannual composites for the years 1984-2014 were used for trend analysis. In order to derive the disturbances only in forests, forest cover layer was aggregated and the original composites were masked. It has been found, that around 23% of forests were disturbed during the study period.

Comparative analysis of Spanish Banks��� Standalone credit profile

The paper studies the relationship between four differently rated bank���s financial profile and their standalone credit rating issued by Moody���s. The comparative analysis shows an example that despite their pricing power and geographical coverage, larger banks do not necessarily have better credit ratings. Instead, business model and risk appetite seem to be the defining factors of banks��� vulnerability to shocks, such as the Spanish real estate crisis. The risk-return relationship is also identified in the banks��� fundamentals meaning that while expansionary strategy in riskier asset classes enhances margins, it also potentially distorts the credit risk profile.

Return on assets in the Tobacco Industry analysis of the four main Tobacco Manufacturers

This paper studies operating profitability drivers in the Four Main Tobacco Manufacturers for the period 2004-2014. The operating profitability is analyzed as return on assets (ROA) based on the DuPont Extended Model breakdown in degree of operational risk, gross sales margin and assets turnover. The sources of ROA are market share and price strategies appraised through the drivers: firm-size, global value and strategic choices. Using consolidated data, results suggest that firm-size and global value holds a positive relationship with ROA. Also innovation through less harmful tobacco products can lead to better ROA despite no correlation between R&D and ROA.

The Arabidopsis PHYTOCHROME KINASE SUBSTRATE2 protein is a phototropin signaling element that regulates leaf flattening and leaf positioning.

In Arabidopsis (Arabidopsis thaliana), the blue light photoreceptor phototropins (phot1 and phot2) fine-tune the photosynthetic status of the plant by controlling several important adaptive processes in response to environmental light variations. These processes include stem and petiole phototropism (leaf positioning), leaf flattening, stomatal opening, and chloroplast movements. The PHYTOCHROME KINASE SUBSTRATE (PKS) protein family comprises four members in Arabidopsis (PKS1-PKS4). PKS1 is a novel phot1 signaling element during phototropism, as it interacts with phot1 and the important signaling element NONPHOTOTROPIC HYPOCOTYL3 (NPH3) and is required for normal phot1-mediated phototropism. In this study, we have analyzed more globally the role of three PKS members (PKS1, PKS2, and PKS4). Systematic analysis of mutants reveals that PKS2 (and to a lesser extent PKS1) act in the same subset of phototropin-controlled responses as NPH3, namely leaf flattening and positioning. PKS1, PKS2, and NPH3 coimmunoprecipitate with both phot1-green fluorescent protein and phot2-green fluorescent protein in leaf extracts. Genetic experiments position PKS2 within phot1 and phot2 pathways controlling leaf positioning and leaf flattening, respectively. NPH3 can act in both phot1 and phot2 pathways, and synergistic interactions observed between pks2 and nph3 mutants suggest complementary roles of PKS2 and NPH3 during phototropin signaling. Finally, several observations further suggest that PKS2 may regulate leaf flattening and positioning by controlling auxin homeostasis. Together with previous findings, our results indicate that the PKS proteins represent an important family of phototropin signaling proteins.

Optimization of Lignin Peroxidase, Manganese Peroxidase, and Lac Production from Ganoderma lucidum Under Solid State Fermentation of Pineapple Leaf

This study was undertaken to isolate ligninase-producing white-rot fungi for use in the extraction of fibre from pineapple leaf agriwaste. Fifteen fungal strains were isolated from dead tree trunks and leaf litter. Ligninolytic enzymes (lignin peroxidase (LiP), manganese peroxidase (MnP), and laccase (Lac)), were produced by solid-state fermentation (SSF) using pineapple leaves as the substrate. Of the isolated strains, the one showing maximum production of ligninolytic enzymes was identified to be Ganoderma lucidum by 18S ribotyping. Single parameter optimization and response surface methodology of different process variables were carried out for enzyme production. Incubation period, agitation, and Tween-80 were identified to be the most significant variables through Plackett-Burman design. These variables were further optimized by Box-Behnken design. The overall maximum yield of ligninolytic enzymes was achieved by experimental analysis under these optimal conditions. Quantitative lignin analysis of pineapple leaves by Klason lignin method showed significant degradation of lignin by Ganoderma lucidum under SSF

Parametric pension reform and the intensive margin of labor supply, evidence from Colombia

We analyze the effect of a parametric reform of the fully-funded pension regime in Colombia on the intensive margin of the labor supply. We take advantage of a threshold defined by law in order to identify the causal effect using a regression discontinuity design. We find that a pension system that increases retirement age and the minimum weeks during which workers must contribute to claim pension benefits causes an increase of around 2 hours on the number of weekly worked hours; this corresponds to 4% of the average number of weekly worked hours or around 14% of a standard deviation of weekly worked hours. The effect is robust to different specifications, polynomial orders and sample sizes.