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Sample records for broccoli coq5 methyltransferase

  1. The effects of Selenocysteine Se-Methyltransferase on Selenium metabolism in Broccoli

    Science.gov (United States)

    Selenium (Se) plays an indispensable role in human nutrition and has suggested to have important health benefits, including as a cancer preventative agent. Se-methylselenocysteine, a monomethylated form of Se, has been shown to be one of the most effective chemopreventative compounds. Broccoli is ...

  2. Aggregates, broccoli and cauliflower

    Science.gov (United States)

    Grey, Francois; Kjems, Jørgen K.

    1989-09-01

    Naturally grown structures with fractal characters like broccoli and cauliflower are discussed and compared with DLA-type aggregates. It is suggested that the branching density can be used to characterize the growth process and an experimental method to determine this parameter is proposed.

  3. Identification of Resistance to Blackrot in Broccoli

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The screening for Black rot(Xanthomonas campestris PV.campestris)resistance in broccoli (Brassica oleracea L.Var.italica Planch) lines plays an important role in breeding for Black rot resistance and is of great value to practical utilization.The broccoli in bred line"91-9318-4"was used as the experimental material in the trial.X0-2 strain of Xanthomonas campestris PV.campestris was and purified from the infected broccoli plants in the field.The spraying method was adopted in the inoculation.The trial was designed as the randomized blocks of two factors with three replications.Factor A was the foliar age and factor B was the concetrations of inoculum.Factor A had three levels A1,A2 and A3 (the 2nd 4th and 6th true-leaf stage),factor B had three levels B1,B2 and B3(106 counts/mL,107 counts/mL and 108 counts/mL).26 breeding materials of broccoli were identified for resistance to Black rot by means of the acquired method.One resistant breeding material was obtained,the identifying results showed that this screening method is practical.

  4. Synthesis of Lysine Methyltransferase Inhibitors

    Science.gov (United States)

    Ye, Tao; Hui, Chunngai

    2015-07-01

    Lysine methyltransferase which catalyze methylation of histone and nonhistone proteins, play a crucial role in diverse biological processes and has emerged as a promising target for the development of various human diseases, including cancer, inflammation, and psychiatric disorders. However, inhibiting Lysine methyltransferases selectively has presented many challenges to medicinal chemists. During the past decade, lysine methyltransferase inhibitors covering many different structural classes have been designed and developed. In this review, we describe the development of selective, small-molecule inhibitors of lysine methyltransferases with an emphasis on their discovery and chemical synthesis. We highlight the current state of lysine methyltransferase inhibitors and discuss future directions and opportunities for lysine methyltransferase inhibitor discovery.

  5. Therapeutic Potential of Brassica oleracea (Broccoli - A Review

    Directory of Open Access Journals (Sweden)

    Chandini Ravikumar

    2015-05-01

    Full Text Available Broccoli is an edible green plant that is classified in the Italica cultivar group of the species Brassica oleracea. They are rich in vitamin C, dietary fiber and also contain glucoraphin, sulforaphane, selenium and isothiocyanates. Broccoli is also an excellent source of indole-3-carbinol. These constituents present in broccoli are known to be very popular since they possess several anti-cancer properties and benefits. These anti-carcinogenic compounds have a wide variety of uses and benefits for the treatment of various diseases and disorders. Broccoli is widely used in the treatment of several forms of cancer and also treats other neural disorders. The therapeutic potential of broccoli has been explained under its role in cancer, diabetes and other diseases. In the treatment of cancer, most of the constituents or the phytochemicals of broccoli such as brassinin, isothiocyanates, indole-3-carbinol etc. have been proved to be effectively beneficial. Even selenium plays a very important role in cancer prevention. The antioxidant activity of broccoli is induced by other phytochemicals such as glucosinolates, glucoraphin and sulforaphane. Sulforaphane in broccoli sprouts also has the potential to cure neural disorders such as Alzheimer’s disease and Parkinson’s disease. It is also used to bring about cure in asthma and diabetic patients. Flavonoids have the effect of reducing the risk of diabetes. Therefore sulforaphane is widely used to treat various diseases and disorders.

  6. Broccoli curves and the tropical invariance of Welschinger numbers

    CERN Document Server

    Gathmann, Andreas; Schroeter, Franziska

    2011-01-01

    In this paper we introduce broccoli curves, certain plane tropical curves of genus zero related to real algebraic curves. The numbers of these broccoli curves through given points are independent of the chosen points - for arbitrary choices of the directions of the ends of the curves, possibly with higher weights, and also if some of the ends are fixed. In the toric Del Pezzo case we show that these broccoli invariants are equal to the Welschinger invariants (with real and complex conjugate point conditions), thus providing a proof of the independence of Welschinger invariants of the point conditions within tropical geometry. The general case gives rise to a tropical Caporaso-Harris formula for broccoli curves which suffices to compute all Welschinger invariants of the plane.

  7. Histone methyltransferases in cancer

    DEFF Research Database (Denmark)

    Albert, Mareike; Helin, Kristian

    2009-01-01

    Cancer is perceived as a heterogeneous group of diseases that is characterized by aberrant patterns of gene expression. In the last decade, an increasing amount of data has pointed to a key role for epigenetic alterations in human cancer. In this review, we focus on a subclass of epigenetic...... regulators, namely histone methyltransferases (HMTs). Several HMTs have been linked to different types of cancer; however, in most cases we only have limited knowledge regarding the molecular mechanisms by which the HMTs contribute to disease development. We summarize the current knowledge regarding some of...

  8. THE EFFECTS OF SELENOCYSTEINE SE-METHYLTRANSFERASE ON SELENIUM METABOLISM IN TRANSGENIC BROCCOLI

    Science.gov (United States)

    Selenium (Se) plays an indispensable role in human nutrition and has been implicated to have important health benefits, including being a cancer preventative agent. Se-methylselenocysteine, a monomethylated form of Se, has been shown to be one of the most effective chemopreventative compounds. Bro...

  9. Plant characters of broccoli determinants of head production

    OpenAIRE

    Fernanda Daniela Brandelero; Betania Brum; Lindolfo Storck; Jessica Cardoso; Talita Slota Kutz; Thiago de Oliveira Vargas

    2016-01-01

    ABSTRACT: The increasing consumption of single-head broccoli is due to several factors, among them there are food production in minimally processing form and the existence of hybrids that adapt to various climates, in addition to the simple harvesting of this typical architecture.This study aimed to identify the most relevant plant characters of broccoli, represented by growth characters, which are determinant in the production and canopy area. The study was conducted in an experimental area ...

  10. Direct-Seeded Broccoli Responses to Reduced Nitrogen Application at Shoot-Tip Straightened Stage

    OpenAIRE

    LI Hong; Li, Tingxian; Robert J. Gordon; Asiedu, Samuel K.

    2009-01-01

    Broccoli (Brassica oleracea var. italica) is an important high-nutritional-value vegetable yet broccoli plant and nitrogen nutrition relations are not well understood. We conducted a study of broccoli plant response to nitrogen nutrient treatments in a commercial production field in Nova Scotia. The objectives were to quantify the effects of nitrogen nutrition on direct-seeded broccoli development and plant nitrogen uptake in different soils. The nitrogen treatments consisted of the rates of ...

  11. Selenium enrichment of broccoli sprout extract increases chemosensitivity and apoptosis of LNCaP prostate cancer cells

    Directory of Open Access Journals (Sweden)

    Suzuki Kazuhiro

    2009-11-01

    Full Text Available Abstract Background Broccoli is a Brassica vegetable that is believed to possess chemopreventive properties. Selenium also shows promise as an anticancer agent. Thus, selenium enrichment of broccoli has the potential to enhance the anticancer properties of broccoli sprouts. Method Selenium-enriched broccoli sprouts were prepared using a sodium selenite solution. Their anticancer properties were evaluated in human prostate cancer cell lines and compared with those of a control broccoli sprout extract. Results Selenium-enriched broccoli sprouts were superior to normal broccoli sprouts in inhibiting cell proliferation, decreasing prostate-specific antigen secretion, and inducing apoptosis of prostate cancer cells. Furthermore, selenium-enriched broccoli sprouts but, not normal broccoli sprouts, induced a downregulation of the survival Akt/mTOR pathway. Conclusion Our results suggest that selenium-enriched broccoli sprouts could potentially be used as an alternative selenium source for prostate cancer prevention and therapy.

  12. Dielectric properties of tantalum powder with broccoli-like morphology

    International Nuclear Information System (INIS)

    Metallic tantalum powder with broccoli-like morphology, consisting of spherical fine particles and of long rods or thin plates, was prepared in a hundred gram scale by calcium reduction of Ta2O5 in molten CaCl2. The properties as electrolytic capacitor were evaluated in comparison with commercial powder obtained by Na reduction and with powder consisting of only fine particles obtained by Ca reduction. The capacitance was larger than that of conventional powder with the same surface area, because the broccoli-like powder showed a strong resistance against shrinkage during high temperature annealing due to the framework of branches. The powder with new broccoli-like morphology can circumvent the conventional treatments for grain size control and gas removal

  13. Genotype and Environment Effects on Carotenoid Content of Broccoli

    Science.gov (United States)

    Carotenoids are secondary plant metabolites in vegetables reported to confer various positive health-promoting effects when consumed. Brassica oleracea L. vegetables are recognized as excellent sources of dietary carotenoids. Broccoli has emerged as the most important B. oleracea crop in the US an...

  14. The Broccoli Syndrome: Higher Education's Pubdown Of The Minority Student

    Science.gov (United States)

    Tate, Penfield; Delworth, Ursula

    1973-01-01

    The assumption underlying the broccoli syndrome is that minority group students are not familiar with a whole range of Anglo goodies''. This article points out how, as the minority student is enlightened time and again to such facts'', he perceives inherent racism in his enlightener.'' (JC)

  15. A transdisciplinary approach to developing an eastern broccoli industry

    Science.gov (United States)

    Current economic, social and scientific situations have made it attractive to establish a substantial and profitable broccoli industry in the eastern US, where several obstacles have prevented that from happening previously. Because many barriers must be overcome simultaneously, we assembled a tran...

  16. SOIL MULCHING WITH STRAW IN BROCCOLI CULTIVATION FOR EARLY HARVEST

    Directory of Open Access Journals (Sweden)

    Edyta Kosterna

    2014-04-01

    Full Text Available All treatments which protect soil from degradation and use of plant protection methods, other than chemicals are of great importance in the cultivation. This effect is attributed, among others, to organic mulches. By limiting the growth of weeds, maintaining proper moisture and reducing daily temperature fluctuations, mulch improves soil conditions for plant growth and development. The experiment was carried out between 2010 and 2012 at the Experimental Farm in Zawady as a split-block design with three replicates. The effect of the kind of straw (rye, corn, rape, buckwheat and its dose (10 and 20 t∙ha-1 applied as a mulch on the yield and quality of broccoli ‘Milady F1’ cultivated for early harvest was investigated. The effect of straw was compared to a control plot without mulch. Weather conditions in the successive years of the study had a significant influence on the yield and quality of broccoli. The highest yields with the best parameters were obtained in 2010 and 2012, which were characterized by sufficient rainfall for broccoli. It was not found significant differences in the yield level and weight of head between particular kinds of straw, however, all kinds of straw investigated in the experiment, irrespective of dose, contributed to a significant increase in the yield and favourably influenced the biometric features of broccoli compared to that achieved from cultivation without straw. Soil mulching with corn straw was most favourable to yield and its parameters.

  17. No de novo sulforaphane biosynthesis in broccoli seedlings

    NARCIS (Netherlands)

    Gorissen, Antonie; Kraut, Nicolai U.; de Visser, Ries; de Vries, Marcel; Roelofsen, Han; Vonk, Roel J.

    2011-01-01

    The isothiocyanate sulforaphane, present in significant amounts in broccoli (Brassica oleracea L.) seedlings in the form of its precursor glucoraphanin, has been identified as an inducer of quinine reductase, a phase-II detoxification enzyme known for its anticarcinogenic properties. Its concentrati

  18. The Effect of Steaming on the Glucosinolate Content in Broccoli

    NARCIS (Netherlands)

    Verkerk, R.; Knol, J.J.; Dekker, M.

    2010-01-01

    Total and individual glucosinolates were measured after different duration of steaming broccoli (Brassica oleracea L. var. italica). During steaming, the temperature profile, cell lysis and inactivation of myrosinase were assessed as well. Steaming resulted in high retention of total aliphatic and i

  19. Innovative minimal processing of mini broccoli for keeping quality and safety and enhancing bioactive compounds

    OpenAIRE

    Martínez Hernández, Ginés Benito

    2012-01-01

    [ENG] Broccoli has been described as the ‘super–vegetable’ in the media after a great number of epidemiological and laboratory studies on this Brassica specie. These studies have shown numerous health–promoting properties of broccoli such as chemopreventive, antioxidant, antitumor, antimutagenicity, anti–inflammatory, antimicrobial, antiviral and reduction of coronary heart disease risk. These properties are related to the high content of broccoli in phenolic compounds, glucosinolates (isothi...

  20. A Market-Window Analysis for Crown-Cut Broccoli Produced in Southwest Virginia

    OpenAIRE

    Dillard, John; Kmak, Heather; Russ, M.J.; Coale, Charles W., Jr.; Bratsch, Tony; Reaves, Dixie Watts

    2006-01-01

    This research determines the profitability for farmers of marketing film-wrapped Crown-Cut broccoli in Southwest Virginia. Historical price data was collected in three eastern terminal markets (Atlanta, Baltimore, and Philadelphia) for Crown-Cut and bunched broccoli from October 1998 to January 2005. Cost-of-production budgets were estimated for Crown-Cut broccoli. The economic data was entered into a market window analysis and the profitability of each market was evaluated. The profitability...

  1. THE INFLUENCE OF NUTRITION ON THE MAIN PRODUCTION OF BROCCOLI

    Directory of Open Access Journals (Sweden)

    Mădălina Doltu

    2013-07-01

    Full Text Available The research was realized in the unheated greenhouse of Horting Institute Bucharest, in 2012 year. It have aimed the influence of one phase of fertilization with ammonium nitrate (NH4NO3 on inflorescences of broccoli, at a hybrid F1 - Ironman. The fertilizer was used before the formation of main inflorescences, in different doses (100 kg/ha, 150 kg/ha, without fertilizer. Harvesting was done in 4 stages. A phase of fertilization with ammonium nitrate has influenced the main production of broccoli: the variant with fertilization 150 kg/ha has obtained the highest percentage of inflorescences formed, the average weight/inflorescence largest and superior production compared with other variants. Were obtained direct linear correlations between the doses of ammonium nitrate (NH4NO3 and the production aspects followed (percentage of main inflorescences harvested, average weight/inflorescence, total production, significances of the correlations very high.

  2. Broccoli, PTEN deletion and prostate cancer: where is the link?

    Directory of Open Access Journals (Sweden)

    Bardelli Alberto

    2010-12-01

    Full Text Available Abstract The concept that vegetables and fruits are relevant sources of cancer-preventive substances is strongly supported by population studies. Among others, cruciferous vegetables like broccoli, cabbage, cauliflower and Brussels sprouts are thought to affect the development of various types of cancers and especially prostate tumors. Yet, the identification of the molecular mechanisms by which the 'active' compounds contained in these vegetables mediate their anticancer activity has historically lagged behind. Accordingly, direct laboratory evidence of how individual nutrients affect cancer genes and the pathways they control remains the major obstacle to progress in this research field. Here we review a recent report investigating the interaction between sulforaphane, a dietary isothiocyanate derived from broccoli, and expression of the PTEN tumor suppressor gene in pre malignant prostate tissue.

  3. Remote sensing of water and nitrogen stress in broccoli

    Science.gov (United States)

    Elsheikha, Diael-Deen Mohamed

    Remote sensing is being used in agriculture for crop management. Ground based remote sensing data acquisition system was used for collection of high spatial and temporal resolution data for irrigated broccoli crop. The system was composed of a small cart that ran back and forth on a rail system that was mounted on a linear move irrigation system. The cart was equipped with a sensor that had 4 discrete wavelengths; 550 nm, 660 nm, 720 nm, and 810 nm, and an infrared thermometer, all had 10 nm bandwidth. A global positioning system was used to indicate the cart position. The study consisted of two parts; the first was to evaluate remotely sensed reflectance and indices in broccoli during the growing season, and determine whether remotely sensed indices or standard deviation of indices can distinguish between nitrogen and water stress in broccoli, and the second part of the study was to evaluate remotely sensed indices and standard deviation of remotely sensed indices in broccoli during daily changes in solar zenith angle. Results indicated that nitrogen was detected using Ratio Vegetation index, RVI, Normalized Difference Vegetation Index, NDVI, Canopy Chlorophyll Concentration Index, CCCI, and also using the reflectance in the Near-Infrared, NIR, bands. The Red reflectance band capability of showing stress was not as clear as the previous indices and bands reflectance. The Canopy Chlorophyll Concentration Index, CCCI, was the most successful index. The Crop Water Stress Index was able to detect water stress but it was highly affected by the solar zenith angle change along the day.

  4. Formation of broccoli-like morphology of tantalum powder

    International Nuclear Information System (INIS)

    'Broccoli'-like morphology of Ta powder was found when Ca reduces Ta2O5 in the molten CaCl2. It consisted of fine particles and branches, and it was different from the conventional spherical particles. The formation of this morphology depended on the stacking methods of the starting materials. Eight types of filling methods proved that the branch was formed when the CaO-enriched region was locally produced near the oxide

  5. Polymer coating, germination and vigor of broccoli seeds

    OpenAIRE

    Almeida Celina de; Rocha Sandra Cristina dos Santos; Razera Luiz Fernandes

    2005-01-01

    Brassica oleracea var italica occupies a special place in the internal and external market of vegetables seeds. Vegetables producers demand seeds with high degree of purity, germination and vigor, since seeds' quality is the basis for the success of the production. In this work, broccoli seeds were coated in a spouted bed, by an aqueous suspension of hidroxy-ethyl-cellulose. Effects of the operating variables: spouting air temperature, atomizing air pressure and coating suspension flow rate o...

  6. Evaluation of genotypic variation of broccoli (brassica oleracea var. italic) in response to selenium treatment

    Science.gov (United States)

    Broccoli (Brassica oleracea var. italic) fortified with selenium (Se) has been promoted as a functional food. Here we evaluated 38 broccoli accessions for their capacity to accumulate Se and for their responses to selenate treatment in terms of nutritional qualities and sulfur gene expression. We fo...

  7. Breeding Vegetables Adapted to High Temperatures: A Case Study with Broccoli

    Science.gov (United States)

    Broccoli (Brassica oleracea L. Italica Group) has emerged in the last few decades as one of the most economically important vegetables grown in the United States. Evidence indicates that head development of most broccoli varieties is arrested by temperatures exceeding 30 C, temperatures that are ne...

  8. NOTICE OF RESEASE OF DOWNY MILDEW RESISTANT GREEN SPROUTING BROCCOLI INBRED LINE USVL089

    Science.gov (United States)

    Downy mildew is one of the most destructive diseases of broccoli and other Brassica oleracea L. crops. This fungal disease, caused by the biotrophic parasite Peronospora parasitica (Pers. Fr.), is a problem worldwide. Fungicide application can provide some control of downy mildew in broccoli. How...

  9. SPECIATION OF SELENIUM COMPOUNDS FROM HIGH SELENIUM BROCCOLI IS AFFECTED BY THE EXTRACTING SOLUTION

    Science.gov (United States)

    The speciation of selenium compounds from high selenium broccoli (876 ¿g/g) depends on the extraction conditions. Twenty-seven extraction conditions were explored involving nine different buffering systems between pH 1 and pH 9. In non-buffered extractions of broccoli, more than 40% of the spiked ...

  10. BIOAVAILABILITY OF SELENIUM FROM MEAT AND BROCCOLI AS DETERMINED BY RETENTION AND DISTRIBUTION OF SE75

    Science.gov (United States)

    Meat is the single greatest source of selenium (Se) in the North American diet. Although not naturally enriched in Se, broccoli will accumulate Se when grown on high Se soils. Previous reports have demonstrated that Se from meat is highly bioavailable whereas Se from broccoli has poor bioavailabil...

  11. Glucosinolate variation among six cultigens of broccoli grown in five diverse east coast locations

    Science.gov (United States)

    Broccoli (Brassica oleracea var. italica) consumption has increased in the United States, driven at least partially by recognition that it is highly nutritious and contains high concentrations of glucosinolates (GS). Glucosinolates are secondary metabolites in broccoli that when digested have a det...

  12. Bioavailability and kinetics of sulforaphane in humans after consumption of cooked versus raw broccoli

    NARCIS (Netherlands)

    Vermeulen, M.; Klöpping-Ketelaars, I.W.A.A.; Berg, R. van den; Vaes, W.H.J.

    2008-01-01

    The aim of this study was to determine the bioavailability and kinetics of the supposed anticarcinogen sulforaphane, the hydrolysis product of glucoraphanin, from raw and cooked broccoli. Eight men consumed 200 g of crushed broccoli, raw or cooked, with a warm meal in a randomized, free-living, open

  13. A research approach for quality based design of healthy foods: Dried broccoli as a case study

    NARCIS (Netherlands)

    Oliviero, T.; Verkerk, R.; Dekker, M.

    2013-01-01

    An approach for process design based on optimization of product quality attributes is presented. Adsorption drying of broccoli with retention of its health benefits is taken as an example to illustrate the approach. Related to its content of glucosinolates, broccoli has a high potential to reduce th

  14. Effect of Hot Water Treatment on Postharvest Shelf Life and Quality of Broccoli

    Institute of Scientific and Technical Information of China (English)

    WU Ping; LI Wu

    2003-01-01

    Broccoli was stored at 0, 10, or 20℃ after immersion in hot water (38 -52℃ ) for 10 or 30min. Yellowing of broccoli was significantly slowed and shelf life significantly increased when broccoli wastreated with hot water at 42 -46℃ and then stored at 10 or 20℃. Heat injury occurred when treatment washigher than 46℃ in some varieties. Broccoli lasted 2 -3 days longer when stored at 10℃ and 1 -2 days longerwhen stored at 20℃ after hot water treatment at 46℃. There was no significant effect of treatment on shelflife after long time storage at 0℃. Weight loss was reduced by hot water treatment and the respiration behav-ior of the broccoli was also changed.

  15. Reproductive fitness of outcrossed hybrids between transgenic broccoli (brassica oleracea) carrying the ipt transgene and conventional varieties of kale, broccoli and cauliflower

    International Nuclear Information System (INIS)

    Pollens are potential carriers for genetically modified crops to transfer genetic materials horizontally to other plants. For phanerogams, pollen viability and cross-compatibility are critical factors for successful outcross hybridization. To evaluate this possibility, this project investigated pollen viability and pod setting rate by comparing broccoli (Brassica oleracea L. var. italica Planck) and broccoli transformed with the isopentenyl transferase (ipt) gene. Both served as pollen donors and four other varieties as pollen receptors to determine outcross rates. For pollen viability, F1 progeny was higher (p?0.05) for the cross of transgenic ipt broccoli with Li Syue significantly by FDA (fluorescein diacetate) assay. Higher successful hybrids were observed for transgenic ipt broccoli with Fu Yue, Li Syue and Green King. As pollen properties, number and grain diameter were significantly larger (p?0.05) in hybrid combinations of transgenic ipt broccoli with Li Syue and Green King significantly (p?0.05). The pod setting rates were higher while transgenic ipt broccoli served as donor plant. These results analyzing pollen properties between transgenic crops with possible outcross candidates would serve as one of those critical strategies for evaluating environmental biosafety issues for transgenic crops. (author)

  16. ADAPTABILITY, FERTILIZER EFFECT AND NUTRITION CONTENT OF VARIOUS BROCCOLI CULTIVAR (Brassica oleracea L. var. italica INTRODUCED IN KOPANG, CENTRAL LOMBOK

    Directory of Open Access Journals (Sweden)

    Zaenul Gafari

    2015-09-01

    Full Text Available This research was aimed to determine adaptability of eight introduced broccoli cultivars in the tropical climate, the effect of fertilizers, as well as determine the content of vitamin A, C and fiber owned by the cultivar. This research was using randomized complete block design ( RCBD with two factors. The first factorsare different broccoli cultivars namely Broccoli 'Lucky', Broccoli 'B - 106', Broccoli 'B-116', Broccoli 'B-112', Broccoli 'Green 2109' F1, Broccoli 'Green calabrase' and Broccoli 'Green Magic 'and Broccoli' Mujur'as a control. The second factors fertilizer of ZA, manure and urea. This research was performed in Kopang, Central Lombok, West Nusa Tenggara, for seven months from December 2014 to Juni 2015.Analysis of the nutrient content was done at the Laboratory of Analytical Chemistry, University of Mataram, Lombok. Parameters observed in this research were plant height, number and weight of leaves, and quality of curd. Data were analyzed by analysis of variance (ANOVA if significantly different then tested further by Duncan's Multiple Range Test (DMRT at 5% level. The results showed all the cultivars able to adapt. Broccoli 'B-106' is a cultivar that has the quality and quantity of curd most excellent and able to adapt to high temperatures. ZA fertilizer application will produce curd weight and diameter greater than that ofmanure or urea. The research also showed that cultivar B-112 has the highest content of vitamin A, B-116 has the highest vitamin C and Green 2109F1 has the highest fibers.

  17. Effect of NaCl treatments on glucosinolate metabolism in broccoli sprouts

    Institute of Scientific and Technical Information of China (English)

    Rong-fang GUO; Gao-feng YUAN; Qiao-mei WANG

    2013-01-01

    To understand the regulation mechanism of NaCl on glucosinolate metabolism in broccoli sprouts,the germination rate,fresh weight,contents of glucosinolates and sulforaphane,as well as myrosinase activity of broccoli sprouts germinated under 0,20,40,60,80,and 100 mmol/L of NaCl were investigated in our experiment.The results showed that glucoerucin,glucobrassicin,and 4-hydroxy glucobrassicin in 7-d-old broccoli sprouts were significantly enhanced and the activity of myrosinase was inhibited by 100 mmol/L of NaCl.However,the total glucosinolate content in 7-d-old broccoli sprouts was markedly decreased although the fresh weight was significantly increased after treatment with NaCl at relatively low concentrations(20,40,and 60 mmol/L).NaCl treatment at the concentration of 60 mmol/L for 5 d maintained higher biomass and comparatively higher content of glucosinolates in sprouts of broccoli with decreased myrosinase activity.A relatively high level of NaCl treatment(100 mmol/L)significantly increased the content of sulforaphane in 7-d-old broccoli sprouts compared with the control.These results indicate that broccoli sprouts grown under a suitable concentration of NaCl could be desirable for human nutrition.

  18. Broccoli consumption and chronic atrophic gastritis among Japanese males: an epidemiological investigation.

    Directory of Open Access Journals (Sweden)

    Sato K

    2004-06-01

    Full Text Available Previous in vitro and animal experiments have shown that sulforaphane, which is abundant in broccoli, inhibits Helicobacter pylori (H. pylori infection and blocks gastric tumor formation. This suggests that broccoli consumption prevents chronic atrophic gastritis (CAG introduced by H. pylori infection and, therefore, gastric cancer. For an epidemiological investigation of the relationship between the broccoli consumption and CAG, a cross-sectional study of 438 male employees, aged 39 to 60 years, of a Japanese steel company was conducted. CAG was serologically determined with serum cut-off values set at pepsinogen I < or = 70 ng/ml and a ratio of serum pepsinogen I/pepsinogen II < or = 3.0. Broccoli consumption (weekly frequency and diet were monitored by using a 31-item food frequency questionnaire. The prevalence of CAG among men who ate broccoli once or more weekly was twice as high as that among men who consumed a negligible amount (P < 0.05. Multiple logistic regression analysis indicated that broccoli consumption once or more weekly significantly increased the risk for CAG (odds ratio, 3.06; 95% confidence interval, 1.12-8.38; P < 0.05, after controlling for age, education, cigarette smoking, and alcohol consumption. The present study failed to show an expected association between frequent broccoli consumption and a low prevalence of CAG.

  19. Changes in Some Chemical Components During Germination of Broccoli Seeds

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    Živilė TARASEVIČIENĖ

    2009-11-01

    Full Text Available Seeds germination to obtain sprouts is a technological method used for many years. During this biological process metabolic enzymes are activated and utilization or synthesis of wide range of chemical compounds occurs in seeds. Cruciferous plants seeds are especially valuable for this purpose because of glucosinolates abundance, the amount of which during germination even increases. The aim of this study was to determine the influence of germination process of broccoli (Brassica oleracea L. var. botrytis Plenck seeds on their chemical composition related to nutritional value, including amino acids content. The seeds were germinated for 24, 72 and 120 h in the dark, at temperature of 25°C. The biggest changes in chemical composition were observed after 72 h of germination. During this process the amount of crude proteins increased, as well as amino acids content. Total amount of conjugated amino acids in non-germinated broccoli seeds was on the level of 181 g kg-1 dm and increased after 72 h of germination to 217 g kg-1 dm, while sudden decrease was observed after 120 h of germination, to the level of 177 g kg-1 dm. Prevailing essential amino acids in non-germinated and germinated seeds were leucine and arginine, while non-essential – glutamic acid.

  20. Improvement of the nutraceutical quality of broccoli sprouts by elicitation.

    Science.gov (United States)

    Natella, Fausta; Maldini, Mariateresa; Nardini, Mirella; Azzini, Elena; Foddai, Maria Stella; Giusti, Anna Maria; Baima, Simona; Morelli, Giorgio; Scaccini, Cristina

    2016-06-15

    Epidemiological studies show an inverse association between Brassica consumption and chronic diseases. Phytochemicals are thought to be beneficial for human health and therefore responsible for this protective effect. Increasing their levels into Brassica food is considered an expedient nutritional strategy that can be achieved through the manipulation of growth conditions by elicitors. In this work we systematically evaluated the influence of treatment with different elicitors (sucrose, mannitol, NaCl, 1-aminocyclopropane-L-carboxylic acid, salicylic acid and methyl jasmonate) on the phytochemical composition of broccoli sprouts. The content of total and single glucosinolates, total phenolic compounds, total flavonoids, total anthocyanins, vitamin C and E and β-carotene was assessed. The exposure to different elicitors produced concentration- and elicitor-dependent specific changes in the content of all the phytochemicals considered. Sucrose, identified as the most effective elicitor by principal component analysis, induced a significant increase of total and specific glucosinolates, vitamin C, total anthocyanins and polyphenols. Sucrose is likely to represent an effective tool to increase the nutritional value of broccoli sprouts. PMID:26868554

  1. Polymer coating, germination and vigor of broccoli seeds

    Directory of Open Access Journals (Sweden)

    Almeida Celina de

    2005-01-01

    Full Text Available Brassica oleracea var italica occupies a special place in the internal and external market of vegetables seeds. Vegetables producers demand seeds with high degree of purity, germination and vigor, since seeds' quality is the basis for the success of the production. In this work, broccoli seeds were coated in a spouted bed, by an aqueous suspension of hidroxy-ethyl-cellulose. Effects of the operating variables: spouting air temperature, atomizing air pressure and coating suspension flow rate over the dependent variables: seeds germination, seeds accelerated aging and the speed of seeds germination in soil, were investigated in a factorial scheme trial. The maximum processing time was 120 min. A totally randomized experiment evaluated and compared seeds germination and vigor of the coated and non-coated seeds. There was no identifiable, pronounced difference on germination of coated and non-coated seeds, accelerated aging of seeds, and speed of seeds germination in the soil. Coating with hidroxy-ethyl-cellulose in the spouted bed did not alter broccoli seeds physiologic quality. The surface of coated seeds presented satisfactory distribution and spreading of the polymer film, uniform and individual coating and homogeneous aspect.

  2. Broccoli (Brassica oleracea var. italica head initiation under field conditions

    Directory of Open Access Journals (Sweden)

    Alina Kałużewicz

    2012-12-01

    Full Text Available A two–year study on the influence of temperature on broccoli head initiation was carried out at the ''Marcelin'' experimental station of the Poznań University of Life Sciences. In each year of the study, plants were planted in the field at four dates. The evaluation of the developmental phase of the broccoli shoot apex was based on the analysis of microscope slides. The date of head initiation was assumed as the day on which the first of the examined apices were found to be at the early generative phase. The plant characteristics (number of leaves, leaf area and stem diameter on the date of initiation were also determined. Variation in length of the period from planting to head initiation was found both between dates of planting and between experimental years. The shortest period from planting to initiation was when the plants were planted in April and June (17-18 days in the first year and the longest one for planting in April in the first year of the study (29 days. The length of the period from planting to head initiation depended on mean daily air temperature. The higher the temperature was, the shorter was the period.

  3. Broccoli/weed/soil discrimination by optical reflectance using neural networks

    Science.gov (United States)

    Hahn, Federico

    1995-04-01

    Broccoli is grown extensively in Scotland, and has become one of the main vegetables cropped, due to its high yields and profits. Broccoli, weed and soil samples from 6 different farms were collected and their spectra obtained and analyzed using discriminant analysis. High crop/weed/soil discrimination success rates were encountered in each farm, but the selected wavelengths varied in each farm due to differences in broccoli variety, weed species incidence and soil type. In order to use only three wavelengths, neural networks were introduced and high crop/weed/soil discrimination accuracies for each farm were achieved.

  4. DNA damage and repair activity after broccoli intake in young healthy smokers

    DEFF Research Database (Denmark)

    Riso, Patrizia; Martini, Daniela; Møller, Peter;

    2010-01-01

    compounds, including smokers. The aim of the study was to evaluate the effect of broccoli intake on biomarkers of DNA damage and repair. Twenty-seven young healthy smokers consumed a portion of steamed broccoli (250 g/day) or a control diet for 10 days each within a crossover design with a washout period...... mRNA expression levels of repair and defence enzymes: 8-oxoguanine DNA glycosylase (OGG1), nucleoside diphosphate linked moiety X-type motif 1 (NUDT1) and heme oxygenase 1 (HO-1). After broccoli consumption, the level of oxidised DNA lesions decreased by 41% (95% confidence interval: 10%, 72%) and...... the resistance to H(2)O(2)-induced DNA strand breaks increased by 23% (95% CI: 13%, 34%). Following broccoli intake, a higher protection was observed in subjects with glutathione S-transferase (GST) M1-null genotype. The expression level and activity of repair enzymes was unaltered. In conclusion...

  5. Untargeted Metabolomics Reveals Predominant Alterations in Lipid Metabolism Following Light Exposure in Broccoli Sprouts

    OpenAIRE

    Mariateresa Maldini; Fausta Natella; Simona Baima; Giorgio Morelli; Cristina Scaccini; James Langridge; Giuseppe Astarita

    2015-01-01

    The consumption of vegetables belonging to the family Brassicaceae (e.g., broccoli and cauliflower) is linked to a reduced incidence of cancer and cardiovascular diseases. The molecular composition of such plants is strongly affected by growing conditions. Here we developed an unbiased metabolomics approach to investigate the effect of light and dark exposure on the metabolome of broccoli sprouts and we applied such an approach to provide a bird’s-eye view of the overall metabolic response af...

  6. Broccoli yield in response to top-dressing fertilization with green manure and biofertilizer

    OpenAIRE

    Gilberto Bernardo de Freitas; Márcio Sousa Rocha; Ricardo Henrique Silva Santos; Letícia Monteiro da Silva Freitas; Leandro de Almeida Resende

    2011-01-01

    The objective of this work was to evaluate the productive performance of broccoli under different top-dressing organic fertilizations. The experiment was conducted under protected cultivation, in a completely randomized design with four replications, with two plants per experimental unit. Broccoli seedlings were produced in a commercial substrate in styrofoam trays. The seedlings were transplanted to plastic pots containing 10.0 L of substrate made up of subsoil and organic compost at the rat...

  7. Compositional and Proteomic Analyses of Genetically Modified Broccoli (Brassica oleracea var. italica) Harboring an Agrobacterial Gene

    OpenAIRE

    Mao-Sen Liu; Miau-Hwa Ko; Hui-Chun Li; Shwu-Jene Tsai; Ying-Mi Lai; You-Ming Chang; Min-Tze Wu; Chen, Long-Fang O.

    2014-01-01

    Previously, we showed improved shelf life for agrobacterial isopentenyltransferase (ipt) transgenic broccoli (Brassica oleracea var. italica), with yield comparable to commercial varieties, because of the protection mechanism offered by molecular chaperones and stress-related proteins. Here, we used proximate analysis to examine macronutrients, chemical and mineral constituents as well as anti-nutrient and protein changes of ipt-transgenic broccoli and corresponding controls. We also prelim...

  8. Compositional and Proteomic Analyses of Genetically Modified Broccoli (Brassica oleracea var. italica) Harboring an Agrobacterial Gene

    OpenAIRE

    Liu, Mao-Sen; Ko, Miau-Hwa; Li, Hui-Chun; Tsai, Shwu-Jene; Lai, Ying-Mi; Chang, You-Ming; Wu, Min-Tze; Chen, Long-Fang O.

    2014-01-01

    Previously, we showed improved shelf life for agrobacterial isopentenyltransferase (ipt) transgenic broccoli (Brassica oleracea var. italica), with yield comparable to commercial varieties, because of the protection mechanism offered by molecular chaperones and stress-related proteins. Here, we used proximate analysis to examine macronutrients, chemical and mineral constituents as well as anti-nutrient and protein changes of ipt-transgenic broccoli and corresponding controls. We also prelimin...

  9. Investigation on the influence of pre-treatments on drying behaviour of broccoli by MRI experiments

    OpenAIRE

    Jin, X.; Sman, van der, R.G.M.; Gerkema, E.; Vergeldt, F.J.; As, van, H.; Straten, van, FE; Boom, R.M.; Boxtel, van, C.

    2011-01-01

    Abstract: Magnetic Resonance Imaging (MRI) allows the monitoring of internal moisture content of food products during drying non-destructively. In an experimental set-up with continuous and controlled hot air supply, the internal moisture distribution of broccoli with different pre-treatments are measured during drying. Moisture distribution, drying rate and shrinkage are compared and analyzed quantitatively. MRI results indicated that for fresh broccoli stalks the moisture content in the cor...

  10. The transfer of 'Polima' cytoplasmic male sterility from oilseed rape (Brassica napus) to broccoli (B. oleracea) by protoplast fusion.

    Science.gov (United States)

    Yarrow, S A; Burnett, L A; Wildeman, R P; Kemble, R J

    1990-08-01

    Protoplast fusion was utilised to transfer Polima type cytoplasmic male sterility (CMS) from Brassica napus, canola cv. Polima Karat (Pol-Karat) to B. oleracea, broccoli, var. "Green Comet". Southern and RFLP analysis confirmed that four cybrids possessed nuclear genomes of broccoli with Polima mitochondria and chloroplasts. A fifth cybrid was a nuclear hybrid between broccoli and Pol-Karat, with Polima mitochondria and chloroplasts of broccoli. The broccoli type cybrids were morphologically similar to "Green Comet", while the hybrid type was an intermediate of the two fusion parents. Flowers on the cybrids were distinctive in that although they possessed a morphology typical of Polima, they had very reduced petals. The broccoli type cybrids exhibited some female fertility, albeit low, establishing potential for F1 hybrid production. PMID:24226699

  11. Cultivation conditions and selenium fertilization alter the phenolic profile, glucosinolate, and sulforaphane content of broccoli.

    Science.gov (United States)

    Robbins, Rebecca J; Keck, Anna-Sigrid; Banuelos, Gary; Finley, John W

    2005-01-01

    Broccoli is a food often consumed for its potential health-promoting properties. The health benefits of broccoli are partly associated with secondary plant compounds that have bioactivity; glucosinolates and phenolic acids are two of the most abundant and important in broccoli. In an effort to determine how variety, stress, and production conditions affect the production of these bioactive components broccoli was grown in the greenhouse with and without selenium (Se) fertilization, and in the field under conventional or organic farming procedures and with or without water stress. High-performance liquid chromatography/mass spectrometry was used to separate and identify 12 primary phenolic compounds. Variety had a major effect: There was a preponderance of flavonoids in the Majestic variety, but hydroxycinnamic esters were relatively more abundant in the Legacy variety. Organic farming and water stress decreased the overall production of phenolics. Se fertilization increased glucosinolates in general, and sulforaphane in particular, up to a point; above that Se fertilization decreased glucosinolate production. Organic farming and water stress also decreased glucosinolate production. These data show environmental and genetic variation in phenolics and glucosinolates in broccoli, and warn that not all broccoli may contain all health-promoting bioactive components. They further show that selection for one bioactive component (Se) may decrease the content of other bioactive components such as phenolics and glucosinolates. PMID:16117613

  12. Compositional and proteomic analyses of genetically modified broccoli (Brassica oleracea var. italica) harboring an agrobacterial gene.

    Science.gov (United States)

    Liu, Mao-Sen; Ko, Miau-Hwa; Li, Hui-Chun; Tsai, Shwu-Jene; Lai, Ying-Mi; Chang, You-Ming; Wu, Min-Tze; Chen, Long-Fang O

    2014-01-01

    Previously, we showed improved shelf life for agrobacterial isopentenyltransferase (ipt) transgenic broccoli (Brassica oleracea var. italica), with yield comparable to commercial varieties, because of the protection mechanism offered by molecular chaperones and stress-related proteins. Here, we used proximate analysis to examine macronutrients, chemical and mineral constituents as well as anti-nutrient and protein changes of ipt-transgenic broccoli and corresponding controls. We also preliminarily assessed safety in mice. Most aspects were comparable between ipt-transgenic broccoli and controls, except for a significant increase in carbohydrate level and a decrease in magnesium content in ipt-transgenic lines 101, 102 and 103, as compared with non-transgenic controls. In addition, the anti-nutrient glucosinolate content was increased and crude fat content decreased in inbred control 104 and transgenic lines as compared with the parental control, "Green King". Gel-based proteomics detected more than 50 protein spots specifically found in ipt-transgenic broccoli at harvest and after cooking; one-third of these proteins showed homology to potential allergens that also play an important role in plant defense against stresses and senescence. Mice fed levels of ipt-transgenic broccoli mimicking the 120 g/day of broccoli eaten by a 60-kg human adult showed normal growth and immune function. In conclusion, the compositional and proteomic changes attributed to the transgenic ipt gene did not affect the growth and immune response of mice under the feeding regimes examined. PMID:25170807

  13. Compositional and Proteomic Analyses of Genetically Modified Broccoli (Brassica oleracea var. italica Harboring an Agrobacterial Gene

    Directory of Open Access Journals (Sweden)

    Mao-Sen Liu

    2014-08-01

    Full Text Available Previously, we showed improved shelf life for agrobacterial isopentenyltransferase (ipt transgenic broccoli (Brassica oleracea var. italica, with yield comparable to commercial varieties, because of the protection mechanism offered by molecular chaperones and stress-related proteins. Here, we used proximate analysis to examine macronutrients, chemical and mineral constituents as well as anti-nutrient and protein changes of ipt-transgenic broccoli and corresponding controls. We also preliminarily assessed safety in mice. Most aspects were comparable between ipt-transgenic broccoli and controls, except for a significant increase in carbohydrate level and a decrease in magnesium content in ipt-transgenic lines 101, 102 and 103, as compared with non-transgenic controls. In addition, the anti-nutrient glucosinolate content was increased and crude fat content decreased in inbred control 104 and transgenic lines as compared with the parental control, “Green King”. Gel-based proteomics detected more than 50 protein spots specifically found in ipt-transgenic broccoli at harvest and after cooking; one-third of these proteins showed homology to potential allergens that also play an important role in plant defense against stresses and senescence. Mice fed levels of ipt-transgenic broccoli mimicking the 120 g/day of broccoli eaten by a 60-kg human adult showed normal growth and immune function. In conclusion, the compositional and proteomic changes attributed to the transgenic ipt gene did not affect the growth and immune response of mice under the feeding regimes examined.

  14. Caffeine synthase and related methyltransferases in plants.

    Science.gov (United States)

    Misako, Kato; Kouichi, Mizuno

    2004-05-01

    Caffeine (1,3,7-trimethylxanthine) is a purine alkaloid present in high concentrations in tea and coffee and it is also found in a number of beverages such as coca cola. It is necessary to elucidate the caffeine biosynthetic pathway and to clone the genes related to the production of caffeine not only to determine the metabolism of the purine alkaloid but also to control the content of caffeine in tea and coffee. The available data support the operation of a xanthosine-->7-methylxanthosine-->7-methylxanthine-->theobromine-->caffeine pathway as the major route to caffeine. Since the caffeine biosynthetic pathway contains three S-adenosyl-L-methionine (SAM) dependent methylation steps, N-methyltransferases play important roles. This review focuses on the enzymes and genes involved in the methylation of purine ring. Caffeine synthase, the SAM-dependent methyltransferase involved in the last two steps of caffeine biosynthesis, was originally purified from young tea leaves (Camellia sinensis). The isolated cDNA, termed TCS1, consists of 1,483 base pairs and encodes a protein of 369 amino acids. Subsequently, the homologous genes that encode caffeine biosynthetic enzymes from coffee (Coffea arabica) were isolated. The recombinant proteins are classified into the three types on the basis of their substrate specificity i.e. 7-methylxanthosine synthase, theobromine synthase and caffeine synthase. The predicted amino acid sequences of caffeine biosynthetic enzymes derived from C. arabica exhibit more than 80% homology with those of the clones and but show only 40% homology with TCS1 derived from C. sinensis. In addition, they share 40% homology with the amino acid sequences of salicylic carboxyl methyltransferase, benzoic acid carboxyl methyltransferase and jasmonic acid carboxyl methyltransferase which belong to a family of motif B' methyltransferases which are novel plant methyltransferases with motif B' instead of motif B as the conserved region. PMID:14977590

  15. Incidence of Bacterial Disease and Yield of Broccoli as Influenced by Different Rain Protectors and Varieties during the Rainy Season in Southern Thailand

    OpenAIRE

    Karistsapol Nooprom; Quanchit Santipracha

    2014-01-01

    This study is mainly focused on evaluating the effects of different rain protectors and broccoli varieties to find out whether rain protector and variety is suitable or not for broccoli production during rainy season. Broccoli was experimented at Prince of Songkla University, Hat Yai, Songkhla, southern Thailand. Study revealed that broccoli growing under plastic sheet and green shade net had lower incidence of soft rot disease (1.62 and 3.75%, respectively) than those grown in open field (13...

  16. Directed evolution of improved zinc finger methyltransferases.

    Directory of Open Access Journals (Sweden)

    Brian Chaikind

    Full Text Available The ability to target DNA methylation toward a single, user-designated CpG site in vivo may have wide applicability for basic biological and biomedical research. A tool for targeting methylation toward single sites could be used to study the effects of individual methylation events on transcription, protein recruitment to DNA, and the dynamics of such epigenetic alterations. Although various tools for directing methylation to promoters exist, none offers the ability to localize methylation solely to a single CpG site. In our ongoing research to create such a tool, we have pursued a strategy employing artificially bifurcated DNA methyltransferases; each methyltransferase fragment is fused to zinc finger proteins with affinity for sequences flanking a targeted CpG site for methylation. We sought to improve the targeting of these enzymes by reducing the methyltransferase activity at non-targeted sites while maintaining high levels of activity at a targeted site. Here we demonstrate an in vitro directed evolution selection strategy to improve methyltransferase specificity and use it to optimize an engineered zinc finger methyltransferase derived from M.SssI. The unusual restriction enzyme McrBC is a key component of this strategy and is used to select against methyltransferases that methylate multiple sites on a plasmid. This strategy allowed us to quickly identify mutants with high levels of methylation at the target site (up to ∼80% and nearly unobservable levels of methylation at a off-target sites (<1%, as assessed in E. coli. We also demonstrate that replacing the zinc finger domains with new zinc fingers redirects the methylation to a new target CpG site flanked by the corresponding zinc finger binding sequences.

  17. Using weighted trait indices to select the best performing broccoli hybrids in multi-site and multi-year trials

    Science.gov (United States)

    Understanding and implementing evaluation data from vegetable trials conducted across multiple years and environments by multiple raters presents numerous challenges. In order to select new broccoli hybrids suitable for eastern production, the SCRI East Coast Broccoli Project has conducted over 32 p...

  18. Growth and yield of broccoli fertilized with doses of velvet bean in greenhouse

    Directory of Open Access Journals (Sweden)

    Ellen Rúbia Diniz

    2015-06-01

    Full Text Available The dose effects of green manure in vegetable crops production are still poorly understood. There are few scientific studies indicate that increasing the dose may influence plants characteristics. The objectives were to evaluate the effect of doses of velvet bean green manure on growth and yield, the partitioning of dry matter and nitrogen (N in plants of broccoli, as well as determine the apparent recovery and physiological efficiency of use N. We established four treatments plus a control. The treatments consisted of four levels of green manure: 0, 3, 6, 9 t ha-1 on a dry matter basis, with the addition of 12 t ha-1 of compost. Doses of green manure applied influenced the growth and yield of broccoli. The distribution of dry matter between plant parts of broccoli is not influenced by the doses of green manure. Doses of green manure influenced the amount of N accumulated in the inflorescence and whole plant of broccoli. The apparent recovery of N from green manure and efficiency of N use by plants of broccoli are positively associated with the applied doses.

  19. Anticancer and Antioxidant Activity of Bread Enriched with Broccoli Sprouts

    Directory of Open Access Journals (Sweden)

    Urszula Gawlik-Dziki

    2014-01-01

    Full Text Available This study is focused on antioxidant and anticancer capacity of bread enriched with broccoli sprouts (BS in the light of their potential bioaccessibility and bioavailability. Generally, bread supplementation elevated antioxidant potential of product (both nonenzymatic and enzymatic antioxidant capacities; however, the increase was not correlated with the percent of BS. A replacement up to 2% of BS gives satisfactory overall consumers acceptability and desirable elevation of antioxidant potential. High activity was especially found for extracts obtained after simulated digestion, which allows assuming their protective effect for upper gastrointestinal tract; thus, the anticancer activity against human stomach cancer cells (AGS was evaluated. A prominent cytostatic response paralleled by the inhibition of AGS motility in the presence of potentially mastication-extractable phytochemicals indicates that phenolic compounds of BS retain their biological activity in bread. Importantly, the efficient phenolics concentration was about 12 μM for buffer extract, 13 μM for extracts after digestion in vitro, and 7 μM for extract after absorption in vitro. Our data confirm chemopreventive potential of bread enriched with BS and indicate that BS comprise valuable food supplement for stomach cancer chemoprevention.

  20. Separation and Purification of Sulforaphane from Broccoli by Solid Phase Extraction

    Directory of Open Access Journals (Sweden)

    Kyung Ho Row

    2011-03-01

    Full Text Available A simple solid-phase extraction (SPE method for the determination of sulforaphane in broccoli has been developed. The optimal conditions were found to be use of a silica SPE cartridge, and ethyl acetate and dichloromethane as washing and eluting solvents, respectively, which could eliminate interferences originating from the broccoli matrix. The extracts were sufficiently clean to be directly injected into high-performance liquid chromatography (HPLC for further chromatographic analysis. Good linearity was obtained from 0.05 to 200 μg/mL (r = 0.998 for sulforaphane with the relative standard deviations less than 3.6%. The mean recoveries of sulforaphane from broccoli were more than 90.8% and the detection limit (S/N = 3:1 was 0.02 μg/mL. The SPE method provides a higher yield of sulforaphane from crude extracts compared to conventional liquid-liquid extraction.

  1. Effect of Different Growth State of Broccoli (Brassica oleracea var. italica) on Low Temperature Induction

    Institute of Scientific and Technical Information of China (English)

    JIANG Xin-mei; LI Yue-fang; YU Xi-hong

    2005-01-01

    In order to study the responding of different growth state on low temperature in Broccoli (Brassica oleracea var. italica), we took prematurity broccoli hybrid as the objects. It was found that growth state was varied under different sowing time , moreover, stem became wider with the increasing of light density at the same leaf age.Seedling age of responding to low temperature vernalization only when they grew five leaves above and with over 3.03± 0.07 centimeter stalk width in "Qingfeng Broccoli 103". The older leaf age was and the stronger plant was, the more sensitive and the shorter demanded duration time on low temperature was, and the shorter time required when the plants entered into critical period of floral bud differentiation.

  2. Growth and yield responses of broccoli cultivars to different rates of nitrogen at western Chitwan, Nepal

    DEFF Research Database (Denmark)

    Giri, Raj Kumar; Sharma, Moha Datta; Shakya, Santa Man;

    2013-01-01

    A field experiment was conducted with the objective to determine the optimum rate of nitrogen (N) fertilizer for effective growth and yield of two varieties of broccoli in southern plain of Nepal. The experiment was laid out with two-factorial completely random block design (RCBD) comprising two...... varieties of broccoli (Calabrese and Green Sprouting) and five N rates (0, 50, 100, 150 and 200 kg ha-1) with three replication in each treatment combinations. The effects of variety and N rate on total curd yield were significant but the interaction effect was non-significant. Green Sprouting produced 11...

  3. Ingestion of broccoli sprouts does not improve endothelial function in humans with hypertension

    DEFF Research Database (Denmark)

    Christiansen, Buris; Bellostas Muguerza, Natalia; Petersen, Atheline Major;

    2010-01-01

    be the case in humans suffering from essential hypertension. METHODS: 40 hypertensive individuals without diabetes and with normal levels of cholesterol were examined. The participants were randomized either to ingest 10 g dried broccoli sprouts, a natural donor of glucosinolates with high in vitro...... groups. Diastolic blood pressure stayed essentially unchanged in both groups, while the systolic blood pressure showed a small non significant decrease (9 mm Hg) in the interventional group from a value of 153 mm Hg at start. CONCLUSION: Daily ingestion of 10 g dried broccoli sprouts does not improve...

  4. Pathogen detection, testing, and control in fresh broccoli sprouts

    Directory of Open Access Journals (Sweden)

    Fahey Jed W

    2006-04-01

    Full Text Available Abstract Background The recent increased interest in consuming green vegetable sprouts has been tempered by the fact that fresh sprouts can in some cases be vehicles for food-borne illnesses. They must be grown according to proper conditions of sanitation and handled as a food product rather than as an agricultural commodity. When sprouts are grown in accordance with the criteria proposed from within the sprout industry, developed by regulatory agencies, and adhered to by many sprouters, green sprouts can be produced with very low risk. Contamination may occur when these guidelines are not followed. Methods A one year program of microbial hold-and-release testing, conducted in concert with strict seed and facility cleaning procedures by 13 U.S. broccoli sprout growers was evaluated. Microbial contamination tests were performed on 6839 drums of sprouts, equivalent to about 5 million consumer packages of fresh green sprouts. Results Only 24 (0.75% of the 3191 sprout samples gave an initial positive test for Escherichia coli O157:H7 or Salmonella spp., and when re-tested, 3 drums again tested positive. Composite testing (e.g., pooling up to 7 drums for pathogen testing was equally sensitive to single drum testing. Conclusion By using a "test-and-re-test" protocol, growers were able to minimize crop destruction. By pooling drums for testing, they were also able to reduce testing costs which now represent a substantial portion of the costs associated with sprout growing. The test-and-hold scheme described herein allowed those few batches of contaminated sprouts to be found prior to packaging and shipping. These events were isolated, and only safe sprouts entered the food supply.

  5. Catechol-O-Methyltransferase Polymorphism and Endometriosis

    OpenAIRE

    Wieser, Fritz; Wenzl, Rene; Tempfer, Clemens; Worda, Christoph; Huber, Johannes; Schneeberger, Christian

    2002-01-01

    Purpose: Catechol-O-methyltransferase (COMT) inactivates the estradiol metabolites, 2-hydroxy and 4-hydroxy catechols, which have been implicated in the pathogenesis of endometriosis. A COMT valine to methionine polymorphism (G-to-A) in exon 4 of the COMT gene is polymorphic in the human population, with 25% of Caucasians being homozygous for the low-activity allele (COMT-L) of the enzyme. In a case-control study we investigated whether this COMT polymorphism is associated with endometriosis.

  6. Structural Chemistry of Human RNA Methyltransferases.

    Science.gov (United States)

    Schapira, Matthieu

    2016-03-18

    RNA methyltransferases (RNMTs) play important roles in RNA stability, splicing, and epigenetic mechanisms. They constitute a promising target class that is underexplored by the medicinal chemistry community. Information of relevance to drug design can be extracted from the rich structural coverage of human RNMTs. In this work, the structural chemistry of this protein family is analyzed in depth. Unlike most methyltransferases, RNMTs generally feature a substrate-binding site that is largely open on the cofactor-binding pocket, favoring the design of bisubstrate inhibitors. Substrate purine or pyrimidines are often sandwiched between hydrophobic walls that can accommodate planar ring systems. When the substrate base is laying on a shallow surface, a 5' flanking base is sometimes anchored in a druggable cavity. The cofactor-binding site is structurally more diverse than in protein methyltransferases and more druggable in SPOUT than in Rossman-fold enzymes. Finally, conformational plasticity observed both at the substrate and cofactor binding sites may be a challenge for structure-based drug design. The landscape drawn here may inform ongoing efforts toward the discovery of the first human RNMT inhibitors. PMID:26566070

  7. Biotechnological advancement in genetic improvement of broccoli (Brassica oleracea L. var. italica), an important vegetable crop.

    Science.gov (United States)

    Kumar, Pankaj; Srivastava, Dinesh Kumar

    2016-07-01

    With the advent of molecular biotechnology, plant genetic engineering techniques have opened an avenue for the genetic improvement of important vegetable crops. Vegetable crop productivity and quality are seriously affected by various biotic and abiotic stresses which destabilize rural economies in many countries. Moreover, absence of proper post-harvest storage and processing facilities leads to qualitative and quantitative losses. In the past four decades, conventional breeding has significantly contributed to the improvement of vegetable yields, quality, post-harvest life, and resistance to biotic and abiotic stresses. However, there are many constraints in conventional breeding, which can only be overcome by advancements made in modern biology. Broccoli (Brassica oleracea L. var. italica) is an important vegetable crop, of the family Brassicaceae; however, various biotic and abiotic stresses cause enormous crop yield losses during the commercial cultivation of broccoli. Thus, genetic engineering can be used as a tool to add specific characteristics to existing cultivars. However, a pre-requisite for transferring genes into plants is the availability of efficient regeneration and transformation techniques. Recent advances in plant genetic engineering provide an opportunity to improve broccoli in many aspects. The goal of this review is to summarize genetic transformation studies on broccoli to draw the attention of researchers and scientists for its further genetic advancement. PMID:26971329

  8. Importance of Genotype on Carotenoid and Chlorophyll Levels in Broccoli Heads

    Science.gov (United States)

    Carotenoids are secondary plant metabolites in vegetables known to be essential in the human diet and reported to confer various positive health-promoting effects when consumed. Brassica oleracea L. vegetables like kale, cabbage, and broccoli are recognized as excellent sources of dietary carotenoi...

  9. Doubled haploid inbred lines USVL048 and USVL131 of heading broccoli

    Science.gov (United States)

    Two inbred lines of heading broccoli (Brassica oleracea L. var. italica), designated USVL048 and USVL131, were released by the Agricultural Research Service of the U.S. Department of Agriculture in 2012. Both of the released lines are doubled haploids originally derived from another culture. As do...

  10. LETTUCE AND BROCCOLI RESPONSE AND SOIL PROPERTIES RESULTING FROM TANNERY WASTE APPLICATIONS

    Science.gov (United States)

    Broccoli (Brassica oleracea L. var. italica) and lettuce (Lactuca sativa L.) were grown on Willamette sil (Pachic Ultic Argixerolls) amended 1 and 2 yr earlier with chrome tannery wastes at rates up to 192 Mg ha to determine nutrient and trace element availability. Soils were sam...

  11. SPECIFIC SELENIUM SPECIES FROM BROCCOLI ARE TRANSPORTED DIFFERENTLY ACROSS A CACO-2 CELL MONOLAYER

    Science.gov (United States)

    Biological actions of selenium (Se) depend in part on its specific chemical form. Reverse-phase HPLC coupled to an Inductively Coupled Plasma Mass Spectrometer (HPLC-ICP-MS) with known Se standards was used to characterize chemical forms of Se in biological matrices. High-Se broccoli contained mul...

  12. Effects of stir-fry cooking with different edible oils on the phytochemical composition of broccoli.

    Science.gov (United States)

    Moreno, Diego A; López-Berenguer, Carmen; García-Viguera, Cristina

    2007-01-01

    Numerous epidemiological studies indicate that Brassica vegetables in general and broccoli in particular protect humans against cancer; they are rich sources of glucosinolates and possess a high content on flavonoids, vitamins, and mineral nutrients. The contents of total intact glucosinolates, total phenolics, vitamin C, and minerals (potassium, sodium, calcium, magnesium, iron, manganese, zinc, and copper) in the edible portions of freshly harvested broccoli (florets), which was subjected to stir-frying treatments, were evaluated. In the present work, the stir-fry cooking experiments were carried out using different edible oils from plant origin (refined olive oil, extra virgin olive oil, sunflower oil, peanut oil, soyabean oil, and safflower oil) known and used worldwide. Results showed that during stir-frying, phenolics and vitamin C were more affected than glucosinolates and minerals. Stir-fry cooking with extra virgin olive, soybean, peanut, or safflower oil did not reduce the total glucosinolate content of the cooked broccoli compared with that of the uncooked sample. The vitamin C content of broccoli stir-fried with extra virgin olive or sunflower oil was similar to that of the uncooked sample, but greater than those samples stir-fried with other oils. PMID:17995900

  13. Broccoli glucosinolate degradation is reduced performing thermal treatment in binary systems with other food ingredients

    NARCIS (Netherlands)

    Giambanelli, E.; Verkerk, R.; Fogliano, V.; Capuano, E.; Antuono, D' L.F.; Oliviero, T.

    2015-01-01

    Glucosinolate (GL) stability has been widely studied in different Brassica species. However, the matrix effect determined by the presence of other ingredients occurred in many broccoli-based traditional recipes may affect GL thermal degradation. In this study, the matrix effect on GL thermal degrada

  14. DEVELOPMENT OF SEQUENCE CHARACTERIZED AMPLIFIED REGION (SCAR) MARKERS LINKED TO DOWNY MILDEW RESISTANCE IN BROCCOLI

    Science.gov (United States)

    Downy mildew, caused by the fungal parasite Peronospora parasitica (Pers.:Fr.) Fr., is one of the most destructive diseases of Brassica oleracea L. crops, including broccoli. As environmental concerns regarding pesticide usage increase, the development and deployment of downy mildew resistant brocc...

  15. Koeling van bloemkool en broccoli op de veiling "De Tuinbouw" te Grootebroek

    NARCIS (Netherlands)

    Damen, P.M.M.; Hendriks, A.G.M.

    1980-01-01

    In een proef op de veiling "De Tuinbouw" in Grootebroek is zowel bloemkool als broccoli opgeslagen gedurende respectievelijk een en drie dagen bij 2 graden C en 95% r.v. Na koeling zijn de kolen geplaatst in een donkere ruimte met een temperatuur van 18 graden C en een r.v. van 90%. Beoordeeld is op

  16. Informatieblad Mineralen en Milieukwaliteit : nitraatuitspoeling uit gewasresten van broccoli, prei en suikerbiet

    NARCIS (Netherlands)

    Ruijter, de F.J.

    2008-01-01

    Om nitraatuitspoeling terug te dringen is het nodig om het N-overschot te verlagen. Bij sommige gewassen (bijv. broccoli en suikerbieten) zit het N-overschot grotendeels in de gewasresten. De vraag is in welke mate het N-overschot in de vorm van gewasresten bijdraagt aan nitraatuitspoeling, en in ho

  17. Broccoli consumption interacts with GSTM1 to perturb oncogenic signalling pathways in the prostate.

    Directory of Open Access Journals (Sweden)

    Maria Traka

    Full Text Available BACKGROUND: Epidemiological studies suggest that people who consume more than one portion of cruciferous vegetables per week are at lower risk of both the incidence of prostate cancer and of developing aggressive prostate cancer but there is little understanding of the underlying mechanisms. In this study, we quantify and interpret changes in global gene expression patterns in the human prostate gland before, during and after a 12 month broccoli-rich diet. METHODS AND FINDINGS: Volunteers were randomly assigned to either a broccoli-rich or a pea-rich diet. After six months there were no differences in gene expression between glutathione S-transferase mu 1 (GSTM1 positive and null individuals on the pea-rich diet but significant differences between GSTM1 genotypes on the broccoli-rich diet, associated with transforming growth factor beta 1 (TGFbeta1 and epidermal growth factor (EGF signalling pathways. Comparison of biopsies obtained pre and post intervention revealed more changes in gene expression occurred in individuals on a broccoli-rich diet than in those on a pea-rich diet. While there were changes in androgen signalling, regardless of diet, men on the broccoli diet had additional changes to mRNA processing, and TGFbeta1, EGF and insulin signalling. We also provide evidence that sulforaphane (the isothiocyanate derived from 4-methylsuphinylbutyl glucosinolate that accumulates in broccoli chemically interacts with TGFbeta1, EGF and insulin peptides to form thioureas, and enhances TGFbeta1/Smad-mediated transcription. CONCLUSIONS: These findings suggest that consuming broccoli interacts with GSTM1 genotype to result in complex changes to signalling pathways associated with inflammation and carcinogenesis in the prostate. We propose that these changes may be mediated through the chemical interaction of isothiocyanates with signalling peptides in the plasma. This study provides, for the first time, experimental evidence obtained in humans to

  18. Mutations in cauliflower and sprout broccoli grown from seeds flown in space

    Science.gov (United States)

    Wu, Hong; Huang, Congli; Zhang, Keping; Sun, Yeqing

    2010-11-01

    Cauliflower and sprout broccoli are widely planted vegetables particularly in Fujian Province, China. To study the mutation in these two types of vegetables induced from spaceflight, we flew the seeds on the 20th Chinese recoverable satellite which orbited the Earth for 18 days. After returning to the Earth, the cauliflower seeds were planted for two generations and the sprout broccoli seeds for one generation at the Xiamen Agriculture Research Institute. Of the 12 cauliflowers planted for the first generation, two showed significant phenotypical changes in both the size of the plant and the weight of the flower head. In addition, most of the space flown plants were found to be resistant to the black rot attack in the field. Cauliflowers planted for the second generation from the seeds in one of the two plants that displayed phenotypical changes in the first generation showed similar mutations. For the first generation of sprout broccoli, the rate of emergence from the flown seeds was lower than that of the control by 30%. No significant changes in the phenotype between the sprout broccolis planted from the flown seeds and the control were observed except one of the mutated sprout broccolis showed a change in the appearance in the lesser bud of the chief flower head. Results of the study demonstrated that DNA damages in some of the genes may have occurred in the seeds flown in space, and some of the changes in the genes may have inherited from the first to the second generation. The improved resistance to the black rot attack and increased size of the flower head are apparently beneficial.

  19. Xylem-to-phloem transfer of boron in broccoli and lupin during early reproductive growth

    International Nuclear Information System (INIS)

    The aim of this study was to test the hypothesis that newly-acquired boron (B) undergoes rapid xylem-to-phloem transfer in plants with restricted mobility. Analysis of the element accumulation and water usage by shoots of intact broccoli (Brassica oleracea var. italica Plenck cv. Commander) and lupin (Lupinus albus L. cv. Ultra) plants provided with a non-deficient supply of B, revealed that the concentration of various mineral elements (K, P, Mg, Ca, B, Fe, Zn, Mo, Cu, Mn) in xylem sap of intact plants ranged from 0.3 μM to 3.5 mM, with B being present at 2.9-3.5 μM. For each element assayed, the concentration was higher in phloem exudate (1.6 μM to 91 mM) than in xylem sap; B was present at about 0.4 mM. Intact broccoli and lupin plants or detached transpiring broccoli shoots were supplied simultaneously with enriched 10B, strontium (a xylem marker) and rubidium (a xylem/phloem marker) during early reproductive growth. The contents of these three compounds were determined in foliage and florets or fruits as a function of time (i.e. up to 12 h and 4 days for broccoli and lupin plants, respectively), and the content in florets or fruits was expressed as a percent of the total recovered. In general, the percent recovery of both 10B and rubidium in florets or fruits was similar and markedly greater than that for strontium, even at the earliest harvest times (within 2 h for broccoli and 1 day for lupin). The data indicate that in plants with restricted B mobility, B is supplied to sink tissues in the phloem, and the extent of B xylem-to-phloem transfer is closely determined by current uptake. (au)

  20. BROCCOLI: Software for Fast fMRI Analysis on Many-Core CPUs and GPUs

    Directory of Open Access Journals (Sweden)

    Anders eEklund

    2014-03-01

    Full Text Available Analysis of functional magnetic resonance imaging (fMRI data is becoming ever more computationally demanding as temporal and spatial resolutions improve, and large, publicly available data sets proliferate. Moreover, methodological improvements in the neuroimaging pipeline, such as non-linear spatial normalization, non-parametric permutation tests and Bayesian Markov Chain Monte Carlo approaches, can dramatically increase the computational burden. Despite these challenges, there do not yet exist any fMRI software packages which leverage inexpensive and powerful graphics processing units (GPUs to perform these analyses. Here, we therefore present BROCCOLI, a free software package written in OpenCL (Open Computing Language that can be used for parallel analysis of fMRI data on a large variety of hardware configurations. BROCCOLI has, for example, been tested with an Intel CPU, an Nvidia GPU and an AMD GPU. These tests show that parallel processing of fMRI data can lead to significantly faster analysis pipelines. This speedup can be achieved on relatively standard hardware, but further, dramatic speed improvements require only a modest investment in GPU hardware. BROCCOLI (running on a GPU can perform non-linear spatial normalization to a 1 mm3 brain template in 4-6 seconds, and run a second level permutation test with 10,000 permutations in about a minute. These non-parametric tests are generally more robust than their parametric counterparts, and can also enable more sophisticated analyses by estimating complicated null distributions. Additionally, BROCCOLI includes support for Bayesian first-level fMRI analysis using a Gibbs sampler. The new software is freely available under GNU GPL3 and can be downloaded from github (https://github.com/wanderine/BROCCOLI/.

  1. BROCCOLI: Software for fast fMRI analysis on many-core CPUs and GPUs.

    Science.gov (United States)

    Eklund, Anders; Dufort, Paul; Villani, Mattias; Laconte, Stephen

    2014-01-01

    Analysis of functional magnetic resonance imaging (fMRI) data is becoming ever more computationally demanding as temporal and spatial resolutions improve, and large, publicly available data sets proliferate. Moreover, methodological improvements in the neuroimaging pipeline, such as non-linear spatial normalization, non-parametric permutation tests and Bayesian Markov Chain Monte Carlo approaches, can dramatically increase the computational burden. Despite these challenges, there do not yet exist any fMRI software packages which leverage inexpensive and powerful graphics processing units (GPUs) to perform these analyses. Here, we therefore present BROCCOLI, a free software package written in OpenCL (Open Computing Language) that can be used for parallel analysis of fMRI data on a large variety of hardware configurations. BROCCOLI has, for example, been tested with an Intel CPU, an Nvidia GPU, and an AMD GPU. These tests show that parallel processing of fMRI data can lead to significantly faster analysis pipelines. This speedup can be achieved on relatively standard hardware, but further, dramatic speed improvements require only a modest investment in GPU hardware. BROCCOLI (running on a GPU) can perform non-linear spatial normalization to a 1 mm(3) brain template in 4-6 s, and run a second level permutation test with 10,000 permutations in about a minute. These non-parametric tests are generally more robust than their parametric counterparts, and can also enable more sophisticated analyses by estimating complicated null distributions. Additionally, BROCCOLI includes support for Bayesian first-level fMRI analysis using a Gibbs sampler. The new software is freely available under GNU GPL3 and can be downloaded from github (https://github.com/wanderine/BROCCOLI/). PMID:24672471

  2. Distinction between the Cfr Methyltransferase Conferring Antibiotic Resistance and the Housekeeping RlmN Methyltransferase

    DEFF Research Database (Denmark)

    Atkinson, Gemma C; Hansen, Lykke H; Tenson, Tanel;

    2013-01-01

    The cfr gene encodes the Cfr methyltransferase that primarily methylates C-8 in A2503 of 23S rRNA in the peptidyl transferase region of bacterial ribosomes. The methylation provides resistance to six classes of antibiotics of clinical and veterinary importance. The rlmN gene encodes the Rlm...

  3. The effect of mulch and fertilizers on broccoli (Brassica oleracea L. Var. Italica oxidants and antioxidants

    Directory of Open Access Journals (Sweden)

    Ali Husain Jasim

    2014-11-01

    Full Text Available Factorial field experiments was conducted during 2012/2013 growth season to study the effect of soil mulch (mulch and without mulch and some fertilizers (control, sulfur, organic and high potash fertilizer on H2O2 content and the activity of catalase (CAT, superoxide dismutase (SOD, and glutathione (GSH oxidative enzymes of broccoli leaves and flowers. The results showed that using mulch, fertilizer treatment and the interaction between mulch and high potash treatment led to a significant increase in CAT and SOD activity. Furthermore, high potash and sulfur supplementation caused a significant increase in broccoli antioxidants both in leaves and flowers respectively. We conclude that mulch and high potash treatment led to increase plant tolerance to salt stress by increasing antioxidant mechanisms.

  4. Neural networks using broadband spectral discriminators reduces illumination required for broccoli identification in weedy fields

    Science.gov (United States)

    Hahn, Federico

    1996-03-01

    Statistical discriminative analysis and neural networks were used to prove that crop/weed/soil discrimination by optical reflectance was feasible. The wavelengths selected as inputs on those neural networks were ten nanometers width, reducing the total collected radiation for the sensor. Spectral data collected from several farms having different weed populations were introduced to discriminant analysis. The best discriminant wavelengths were used to build a wavelength histogram which selected the three best spectral broadbands for broccoli/weed/soil discrimination. The broadbands were analyzed using a new single broadband discriminator index named the discriminative integration index, DII, and the DII values obtained were used to train a neural network. This paper introduces the index concept, its results and its use for minimizing artificial lightning requirements with broadband spectral measurements for broccoli/weed/soil discrimination.

  5. Effects of different cooking methods on health-promoting compounds of broccoli

    Institute of Scientific and Technical Information of China (English)

    Gao-feng YUAN; Bo SUN; Jing YUAN; Qiao-mei WANG

    2009-01-01

    The effects of five domestic cooking methods, including steaming, microwaving, boiling, stir-frying, and stir-frying followed by boiling (stir-frying/boiling), on the nutrients and health-promoting compounds of broccoli were investigated. The results show that all cooking treatments, except steaming, caused significant losses of chlorophyll and vitamin C and significant decreases of total soluble proteins and soluble sugars. Total aliphatic and indole glucosinolates were significantly modified by all cooking treatments but not by steaming. In general, the steaming led to the lowest loss of total glucosinolates, while stir-frying and stir-frying/boiling presented the highest loss. Stir-frying and stir-frying/boiling, the two most popular methods for most homemade dishes in China, cause great losses of chlorophyll, soluble protein, soluble sugar, vitamin C, and glucosinolates, but the steaming method appears the best in retention of the nutrients in cooking broccoli.

  6. 出版业经济惨淡,Broccoli Books关门

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    据出版动画、漫画、游戏类图书的Broccoli Books有限公司总裁Kaname Tezuka披露,Broccoli国际出版美国公司,包括旗下的Broccoli Books出版社和Boysen berry出版社将于今年底关门。他表示,这是日本母公司的决定,“根据美国漫画图书市场和美国经济的整体状况,公司领导层决定最好暂时将力量集中在日本市场。”

  7. Effect of Nitrogen and Boron in Seed Yield and Yield Attributing Characters of Broccoli

    Directory of Open Access Journals (Sweden)

    A. Khanal

    2015-09-01

    Full Text Available Plant nutrient is one of the limiting factors affecting crop production. Nitrogen and boron are major nutrients in case of broccoli. So, an experiment was carried out to evaluate the effect of nitrogen and boron in seed yield and yield attributing characters of broccoli in Rampur, Chitwan during winter season. The experiment was laid out in factorial RCBD design with four levels of nitrogen and two levels of boron. Each plot consists of 25 plants which were separated by 60 * 60 cm spacing. There are altogether eight treatments replicates thrice. Local variety Calabrese was used. Significant effect of different dose of nitrogen and boron on yield attributing characters was found. Also interactive effect of nitrogen and boron in number of pods, pod length, seed yield and number of seeds per pod was found significantly different.

  8. Ozone effects on yield quality of spring oilseed rape and broccoli

    Science.gov (United States)

    Vandermeiren, Karine; De Bock, Maarten; Horemans, Nele; Guisez, Yves; Ceulemans, Reinhart; De Temmerman, Ludwig

    2012-02-01

    The impact of elevated tropospheric ozone (O 3) on the quality of spring oilseed rape ( Brassica napus cv Ability) and broccoli ( Brassica oleracea L. cv Italic cv Monaco) was assessed during a three year Open - Top Chamber (OTC) experiment. Current ambient O 3 levels were compared to an increase of 20 and 40 ppb during 8 h per day over the entire growing season. The qualitative responses were expressed as a function of the accumulated hourly O 3 concentrations over a threshold of 40 ppb (AOT40) and the phytotoxic O 3 dose above a threshold of 6 nmol s -1 m -2 projected leaf area (POD 6). Our results provide clear evidence that O 3 has an influence on the qualitative attributes of the harvested products of these Brassica species. The responses were comparable whether they were expressed as a function of the accumulated O 3 concentrations or of the modelled O 3 uptake. The protein concentration of oilseed rape seeds and broccoli heads was significantly increased in response to O 3. There was also a shift in the fatty acid composition of the vegetable oil derived from seeds of oilseed rape. Oleic acid (18:1) declined significantly ( p rape seeds. In broccoli an important shift occurred from indolic to aliphatic GSLs although the total GSL concentration was not changed. The increase in the aliphatic/indolic GSL ratio ( p < 0.001) may be important in relation to the anticarcinogenic properties of these vegetables. The vitamin C (ascorbate - ASC) and α-TOC concentrations of broccoli were not influenced by O 3; glutathione (GSH) was slightly increased in response to a higher O 3 uptake ( p < 0.05). The consequences of these changes with regard to food and feed quality and human health are discussed.

  9. Multiwalled carbon nanotubes enter broccoli cells enhancing growth and water uptake of plants exposed to salinity

    OpenAIRE

    Martínez-Ballesta, Mª Carmen; Zapata, Lavinia; Chalbi, Najla; Carvajal, Micaela

    2016-01-01

    Background Carbon nanotubes have been shown to improve the germination and growth of some plant species, extending the applicability of the emerging nano-biotechnology field to crop science. Results In this work, exploitation of commercial multiwalled carbon nanotubes (MWCNTs) in control and 100 mM NaCl-treated broccoli was performed. Transmission electron microscopy demonstrated that MWCNTs can enter the cells in adult plants with higher accumulation under salt stress. Positive effect of MWC...

  10. Belowground induction by delia radicum or phytohormones affect aboveground herbivore communities on field-grown broccoli

    OpenAIRE

    Pierre, S.P.; Dugravot, S.; Hervé, M. R.; Hassan, H M; Dam, N.M. van; Cortesero, A. M.

    2013-01-01

    Induced plant defence in response to phytophagous insects is a well described phenomenon. However, so far little is known about the effect of induced plant responses on subsequently colonizing herbivores in the field. Broccoli plants were induced in the belowground compartment using (i) infestation by the root-herbivore Delia radicum, (ii) root application of jasmonic acid (JA) or (iii) root application of salicylic acid (SA). The abundance of D. radicum and six aboveground herbivores display...

  11. Pectobacterium carotovorum subsp. carotovorum – the causal agent of broccoli soft rot in Serbia

    OpenAIRE

    Katarina Gašić; Veljko Gavrilović; Nenad Dolovac; Nenad Trkulja; Svetlana Živković; Danijela Ristić; Aleksa Obradović

    2014-01-01

    Soft rot symptoms were observed on broccoli plants in several commercial fields in the western part of Serbia. Six strains of bacteria were isolated from diseased tissues and identified as Pectobacterium carotovorum subsp. carotovorum using conventional bacteriological and molecular methods. All strains were non-fluorescent, gram-negative, facultative anaerobes, oxidase-negative and catalase-positive, causing soft rot on potato and carrot slices and did not...

  12. Post Harvest Treatment Effects on Crown-Cut Broccoli Shelf Life

    OpenAIRE

    Ramesh, Reddy Yettella Venkata

    2004-01-01

    The effects of packaging treatments, post harvest cooling delay and storage duration on color, texture, ascorbic acid content, weight loss and glucosinolate retention in crown-cut heads of broccoli were studied. Oxygen and CO2 levels inside shrink wrap packages were also monitored. Shrink wrap packaging had a significant positive effect on hue angle (p<0.05). Packaging and post harvest cooling delay had no effect on hue difference and total color difference. While post harvest cooling dela...

  13. Organic broccoli production on transition soils: Comparing cover crops, tillage and sidedress N

    OpenAIRE

    Schellenberg, D.L.; Morse, R.D.; Welbaum, G.E.

    2009-01-01

    Metadata only record Little information is available about how farmers in transition to organic practices should manage short- and long-term N fertility. The objectives of this research were (1) to evaluate the leguminous cover crops lablab (Dolichos lablab L.), soybean (Glycine max L.), sunn hemp (Crotalaria juncea L.) and a mixture of sunn hemp and cowpea (Vigna sinensis Endl.) as N sources; (2) to compare N availability and broccoli yield when cover crops were incorporated with conventi...

  14. Untargeted Metabolomics Reveals Predominant Alterations in Lipid Metabolism Following Light Exposure in Broccoli Sprouts

    Directory of Open Access Journals (Sweden)

    Mariateresa Maldini

    2015-06-01

    Full Text Available The consumption of vegetables belonging to the family Brassicaceae (e.g., broccoli and cauliflower is linked to a reduced incidence of cancer and cardiovascular diseases. The molecular composition of such plants is strongly affected by growing conditions. Here we developed an unbiased metabolomics approach to investigate the effect of light and dark exposure on the metabolome of broccoli sprouts and we applied such an approach to provide a bird’s-eye view of the overall metabolic response after light exposure. Broccoli seeds were germinated and grown hydroponically for five days in total darkness or with a light/dark photoperiod (16 h light/8 h dark cycle. We used an ultra-performance liquid-chromatography system coupled to an ion-mobility, time-of-flight mass spectrometer to profile the large array of metabolites present in the sprouts. Differences at the metabolite level between groups were analyzed using multivariate statistical analyses, including principal component analysis and correlation analysis. Altered metabolites were identified by searching publicly available and in-house databases. Metabolite pathway analyses were used to support the identification of subtle but significant changes among groups of related metabolites that may have gone unnoticed with conventional approaches. Besides the chlorophyll pathway, light exposure activated the biosynthesis and metabolism of sterol lipids, prenol lipids, and polyunsaturated lipids, which are essential for the photosynthetic machinery. Our results also revealed that light exposure increased the levels of polyketides, including flavonoids, and oxylipins, which play essential roles in the plant’s developmental processes and defense mechanism against herbivores. This study highlights the significant contribution of light exposure to the ultimate metabolic phenotype, which might affect the cellular physiology and nutritional value of broccoli sprouts. Furthermore, this study highlights the

  15. Broccoli yield in response to top-dressing fertilization with green manure and biofertilizer

    Directory of Open Access Journals (Sweden)

    Gilberto Bernardo de Freitas

    2011-10-01

    Full Text Available The objective of this work was to evaluate the productive performance of broccoli under different top-dressing organic fertilizations. The experiment was conducted under protected cultivation, in a completely randomized design with four replications, with two plants per experimental unit. Broccoli seedlings were produced in a commercial substrate in styrofoam trays. The seedlings were transplanted to plastic pots containing 10.0 L of substrate made up of subsoil and organic compost at the ratio of 3:1 (v/v, respectively, which is equivalent to about 20.0 t ha-1 of organic compost at planting. After seedling establishment, the top-dressing fertilization treatments were applied: gliricidia biomass associated or not with liquid biofertilizer of cattle manure to the soil and bokashi. Two control treatments were established: one with mineral fertilization recommended for the crop and the other without top-dressing fertilization. The broccoli production was evaluated (commercial standard. Plants that received mineral fertilizer were more productive, however, they were not significantly different (p>0.05, by Dunnet test, from the plants fertilized with 2.5 t ha-1 gliricidiabiomass (dry mass associated with liquid biofertilizer (2.0 L m-2 applied to soil. Top-dressing fertilizations with only gliricidia, at 2.5 and 5.0 t ha-1 of biomass (dry mass, resulted in no significant increase in production of broccoli inflorescence. The use of bokashi in addition to gliricidia biomass and liquid biofertilizer reduced the efficiency of the fertilization compared with plants that received only gliricidia and liquid biofertilizer.

  16. Dark-chilling of Seedlings Affects Initiation and Morphology of Broccoli Inflorescence

    Directory of Open Access Journals (Sweden)

    Aneta GRABOWSKA

    2013-05-01

    Full Text Available The constant supply of broccoli heads for a vegetable market can be kept by the differentiation of sowing time and the age of seedlings. Dark-chilling of young seedlings can also modify the time of transplanting, subsequent plant development, yield and inflorescence quality. The aim of the present study was to analyze the influence of seedling dark-chilling on the inflorescence initiation and the morphological quality of broccoli heads. The experiment was carried out at the University of Agriculture in Krakow (South Poland in the years 2009 and 2010, with broccoli (Brassica oleracea L. var. italica Plenck medium late cultivar ‘Monaco’ F1. The examined factors were the age of seedlings (4-, 6-, 8-, and 10-week-old and dark-chilling (2°C/2 weeks and non-chilled control. Microscopic analysis of the apex sections was performed to determine the time of the inflorescence differentiation in chilled and non-chilled seedlings of different age. Chosen morphological features of the mature heads were also described as depended on age and chilling of seedlings. The inflorescence initiation was observed between 4 and 6 week of the plant ontogeny. Especially 4-week-old seedlings, chilled by two weeks, were more advanced in the generative development in comparison to non-chilled control. Low temperature applied in the seedling stage did not cause the head buttoning. Inflorescences developed by broccoli planted from 4 and 6-week-old seedlings, both chilled and control, were characterized by the best visual quality, determined by a convex head shape, tight branching angle, small and uniform size and regular form of flower buds. Chilling of oldest, 8- and 10-week-old seedlings negatively affected the inflorescence quality, through planed and spread shape of heads and non-uniform shape of flower buds.

  17. Growth and Yield of Broccoli Planted Year Round in Songkhla Province, Thailand

    Directory of Open Access Journals (Sweden)

    Karistsapol Nooprom

    2014-05-01

    Full Text Available Four broccoli (Brassica oleracea var. italica varieties were evaluated in year round of Songkhla province, Thailand, from January, 2011 to January, 2012. The results shown that broccoli planting in all months provided high survival rate of seedling (78.90-98.24%. They had higher plant height than other months when planting in January, February, March, July and December (49.58-51.71 cm. Broccoli planting in January, February and March had the early time to 50% flowering and harvest of 33-36 and 43-45 Days after Transplanting (DAT, respectively. The Yok Kheo could respond to the best seedling survival rate and plant height while the Green Queen had the early time to 50% flowering and harvest. The Yok Kheo planted in January had the highest total yield (12.31 t/ha, followed by the Yok Kheo planted in March (10.65 t/ha and the Top Green planted in January (10.50 t/ha, respectively. The Yok Kheo is a new interesting hybrid variety because it had higher total yield than the Top Green that is the popular variety of farmers in southern Thailand.

  18. BROCCOLI Spears Yield Affected By GAMMA Rays Irradiated Seeds And Foliar Application Of Some Growth Regulators

    International Nuclear Information System (INIS)

    Two field experiments were carried out during 2004/2005 and 2005/2006 winter growing seasons at the experimental farm of Nuclear Research Centre, Atomic Energy Authority, Inshas, Egypt.The experiments were conducted to study the effect of pre-sowing broccoli seeds (cv. F1 175) irradiated with different doses of gamma rays (2, 3 and 4 Gy). The plants were sprayed with GA3 at rate of 50 ml/liter/fed and 20 ml/liter/fed for NAA. Main spear fresh and dry weight per plant, total spears fresh and dry weight per plant, total spears yield, ascorbic acid, TSS, carbohydrates, total chlorophyll, NPK and total protein content of spears were evaluated. The results showed that broccoli seeds irradiated with gamma rays up to 4 Gy pre-sowing increased the abovementioned parameters with different magnitudes comparing with the non-irradiated control plants except spears N, P and protein contents showed decrease in their values comparing with un-treated plants.It could be concluded that the foliar application of GA3 and NAA on broccoli spears increased all the abovementioned parameters, except spears N, P and protein contents showed decrease in their values.

  19. Utility of the broccoli sign in the distinction of prolapsed uterine tumor from cervical tumor

    International Nuclear Information System (INIS)

    Objective: To describe the utility, histopathological basis, and clinical correlates of the broccoli sign. Methods: The committee on human research approved this HIPAA compliant study and waived written informed consent. Based on the records of the senior author and our multidisciplinary Gynecologic Oncology Tumor Board, we retrospectively identified thirteen women (mean age of 48.8 years; range, 34–74) with a cervical mass seen at MR imaging (n = 13) or CT (n = 5) that demonstrated the previously reported broccoli sign (i.e., a soft tissue stalk connecting the cervical mass to the uterine cavity) on one or other modality. All available clinical, imaging, and histopathological records were reviewed, with particular emphasis on initially suspected diagnosis, final proven diagnosis, and outcome. Results: Cervical cancer was the initial clinically suspected diagnosis in 6 of 13 patients. Surgical resection demonstrated prolapsed uterine tumor in all patients, consisting of endometrioid adenocarcinoma (n = 7), carcinosarcoma (n = 2), adenosarcoma (n = 1), and leiomyoma (n = 3). Excluding the three patients with leiomyomas, currently, 7 patients with malignant tumors are disease free after a mean interval of 15 months (range, 3–45) and 3 patients have been lost to follow-up. Conclusion: A stalk connecting an apparent cervical mass seen at CT or MR imaging to the endometrial cavity (“broccoli sign”) favors the diagnosis of a prolapsed uterine tumor; these prolapsed uterine tumors can often be malignant but appear to have a good prognosis.

  20. Effects of broccoli extract on biodistribution and labeling blood components with 99mTc-GH

    International Nuclear Information System (INIS)

    Purpose: people consume vegetables without the knowledge of the side effects of the biological and chemical contents and interactions between radiopharmaceuticals and herbal extract. To this end, current study is focused on the effects of broccoli extract on biodistribution of radiolabeled glucoheptonate (99mTc-GH) and radiolabeling of blood components. Methods: GH was labeled with 99mTc. Quality control studies were done utilizing TLC method. Biodistribution studies were performed on male rats which were treated via gavage with either broccoli extract or SF as control group for 15 days. Blood samples were withdrawn from rats' heart. Radiolabeling of blood constituents performed incubating with GH, SnCl2 and 99m Tc. Results: radiochemical yield of 99mTc-GH is 98.46±1.48 % (n=8). Biodistribution studies have shown that according to the control, the treated group with broccoli has approximately 10 times less uptake in kidney. The percentage of the radioactivity ratios of the blood components is found to be same in both groups. Conclusions: although there is no considerable effect on the radiolabeling of blood components, there is an outstanding change on the biodistribution studies especially on kidneys. The knowledge of this change on kidney uptake may contribute to reduce the risk of misdiagnosis and/or repetition of the examinations in Nuclear Medicine. (author)

  1. Exergetic performance analyses of drying of broccoli florets in a tray drier

    International Nuclear Information System (INIS)

    At present, the drying process is one of the major procedures of food preservation and an important unit operation in a wide variety of food industries. Recently, drying of vegetables is of a particular interest because it is added to various ready-to-eat meals in order to improve their nutritional quality due to health benefit compounds present in vegetables (vitamins, phytochemicals, dietary fibers). Broccoli has been described as a vegetable with a high nutritional value due to its important content of vitamins, antioxidants and anti-carcinogenic compounds. Broccoli dehydration has not been investigated to a great extent and a few data are available in the open literature. In this study, broccoli florets were dried in a tray drier at a temperature range of 50-70 deg C with an air velocity range of 0.5-1.5 m/s. The performance of the process and system was evaluated using the exergy analysis method. Based on the experimental data, effects of the drying air temperature and the velocity on the performance of the drying process were discussed. It was obtained that the exergy evaporation rate and the exergetic efficiency of the process were obtained to vary between 0.0006-0.0029 kW and 0.27-1.16%, respectively. They increased as the drying air temperature increased, while the exergetic efficiency decreased with the rise in the drying air velocity. (author)

  2. Validation of the Analytical Method for the Determination of Flavonoids in Broccoli

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    Tuszyńska Magdalena

    2014-09-01

    Full Text Available A simple, accurate and selective HPLC method was developed and validated for determination of quercetin and kaempferol, which are the main flavonols in broccoli. The separation was achieved on a reversed-phase C18 column using a mobile phase composed of methanol/water (60/40 and phosphoric acid 0.2% at a flow rate of 1.0 ml min-1. The detection was carried out on a DAD detector at 370 nm. This method was validated according to the requirements for new methods, which include selectivity, linearity, precision, accuracy, limit of detection and limit of quantitation. The current method demonstrates good linearity, with R2 > 0.99. The recovery is within 98.07-102.15% and 97.92-101.83% for quercetin and kaempferol, respectively. The method is selective, in that quercetin and kaempferol are well separated from other compounds of broccoli with good resolution. The low limit of detection and limit of quantitation of quercetin and kaempferol enable the detection and quantitation of these flavonoids in broccoli at low con–centrations.

  3. Utility of the broccoli sign in the distinction of prolapsed uterine tumor from cervical tumor

    Energy Technology Data Exchange (ETDEWEB)

    Jha, Priyanka; Chang, Stephanie T. [Department of Radiology, University of California San Francisco, 505 Parnassus Avenue, San Francisco, CA 94143-0628 (United States); Rabban, Joseph T. [Department of Anatomic Pathology, University of California San Francisco, 505 Parnassus Avenue, San Francisco, CA 94143-0628 (United States); Chen, Lee-may [Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Francisco, 505 Parnassus Avenue, San Francisco, CA 94143-0628 (United States); Yeh, Benjamin M. [Department of Radiology, University of California San Francisco, 505 Parnassus Avenue, San Francisco, CA 94143-0628 (United States); Coakley, Fergus V., E-mail: Fergus.Coakley@radiology.ucsf.edu [Department of Radiology, University of California San Francisco, 505 Parnassus Avenue, San Francisco, CA 94143-0628 (United States)

    2012-08-15

    Objective: To describe the utility, histopathological basis, and clinical correlates of the broccoli sign. Methods: The committee on human research approved this HIPAA compliant study and waived written informed consent. Based on the records of the senior author and our multidisciplinary Gynecologic Oncology Tumor Board, we retrospectively identified thirteen women (mean age of 48.8 years; range, 34-74) with a cervical mass seen at MR imaging (n = 13) or CT (n = 5) that demonstrated the previously reported broccoli sign (i.e., a soft tissue stalk connecting the cervical mass to the uterine cavity) on one or other modality. All available clinical, imaging, and histopathological records were reviewed, with particular emphasis on initially suspected diagnosis, final proven diagnosis, and outcome. Results: Cervical cancer was the initial clinically suspected diagnosis in 6 of 13 patients. Surgical resection demonstrated prolapsed uterine tumor in all patients, consisting of endometrioid adenocarcinoma (n = 7), carcinosarcoma (n = 2), adenosarcoma (n = 1), and leiomyoma (n = 3). Excluding the three patients with leiomyomas, currently, 7 patients with malignant tumors are disease free after a mean interval of 15 months (range, 3-45) and 3 patients have been lost to follow-up. Conclusion: A stalk connecting an apparent cervical mass seen at CT or MR imaging to the endometrial cavity ('broccoli sign') favors the diagnosis of a prolapsed uterine tumor; these prolapsed uterine tumors can often be malignant but appear to have a good prognosis.

  4. Bioactive glucosinolates and antioxidant properties of broccoli seeds cultivated in Thailand

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    Sarunya Chuanphongpanich

    2006-03-01

    Full Text Available One of the most significant health concerns of cruciferous vegetables is the presence of biologically active compounds, glucosinolates. Broccoli (Brassica oleracea var. italica is a nutritionally important crop grown all over the world. Glucosinolates have been found to have anti-cancer properties. The primary purpose of this study was to evaluate glucosinolate content and antioxidant property in the seeds of broccoli cultivars widely grown in Thailand. Glucosinolates were analyzed with high performance liquid chromatography (HPLC. Total and individual glucosinolate levels varied significantly among cultivars. In all broccoli seeds, 4-methylsulfinylbutylglucosinolate (glucoraphanin was the predominant glucosinolate. The highest total glucosinolates was 65.5 µmol/g DW in ‘Top Green #067’ cultivar, followed by ‘Packman’ (58.6, ‘Green Queen’ (51.2, ‘Pak Ging’ (25.5 and ‘Rod Fai’ (20.3. The antioxidant capacities, including ABTS radical scavenging activity and ferrous ion chelating ability in the methanol and water extracts, were found to be high.

  5. Detection of the Diversity of Cytoplasmic Male Sterility Sources in Broccoli (Brassica Oleracea var. Italica) Using Mitochondrial Markers

    Science.gov (United States)

    Shu, Jinshuai; Liu, Yumei; Li, Zhansheng; Zhang, Lili; Fang, Zhiyuan; Yang, Limei; Zhuang, Mu; Zhang, Yangyong; Lv, Honghao

    2016-01-01

    Broccoli (Brassica oleracea var. italica) is an important commercial vegetable crop. As part of an efficient pollination system, cytoplasmic male sterility (CMS) has been widely used for broccoli hybrid production. Identifying the original sources of CMS in broccoli accessions has become an important part of broccoli breeding. In this study, the diversity of the CMS sources of 39 broccoli accessions, including 19 CMS lines and 20 hybrids, were analyzed using mitochondrial markers. All CMS accessions contained the ogu orf138-related DNA fragment and the key genes of nap CMS, pol CMS, and tour CMS were not detected. The 39 CMS accessions were divided into five groups using six orf138-related and two simple sequence repeat markers. We observed that ogu CMS R3 constituted 79.49% of the CMS sources. CMS6 and CMS26 were differentiated from the other accessions using a specific primer. CMS32 was distinguished from the other accessions based on a 78-nucleotide deletion at the same locus as the orf138-related sequence. When the coefficient was about 0.90, five CMS accessions (13CMS6, 13CMS23, 13CMS24, 13CMS37, and 13CMS39) exhibiting abnormal floral organs with poor seed setting were grouped together. The polymerase chain reaction amplification profiles for these five accessions differed from those of the other accessions. We identified eight useful molecular markers that can be used to detect CMS types during broccoli breeding. Our data also provide important information relevant to future studies on the possible origins and molecular mechanisms of CMS in broccoli. PMID:27446156

  6. Dietary chemoprevention of PhIP induced carcinogenesis in male Fischer 344 rats with tomato and broccoli.

    Directory of Open Access Journals (Sweden)

    Kirstie Canene-Adams

    Full Text Available The heterocyclic amine, 2-amino-1-methyl-6-phenylimidazo[4,5-B]pyridine (PhIP, found in meats cooked at high temperatures, has been implicated in epidemiological and rodent studies for causing breast, prostate, and colorectal cancers. A previous animal study using a xenograft model has shown that whole tomato and broccoli, when eaten in combination, exhibit a marked effect on tumor reduction compared to when eaten alone. Our aim was to determine if PhIP-induced carcinogenesis can be prevented by dietary consumption of whole tomato + broccoli powders. Male Fischer 344 rats (n = 45 were randomized into the following treatment groups: control (AIN93G diet, PhIP (200 ppm in AIN93G diet for the first 20 weeks of the study, or tomato + broccoli + PhIP (mixed in AIN93G diet at 10% each and fed with PhIP for 20 weeks, and then without PhIP for 32 weeks. Study animals were monitored for 52 weeks and were euthanized as necessary based on a set of criteria for health status and tumor burden. Although there appeared to be some hepatic and intestinal toxicity due to the combination of PhIP and tomato + broccoli, these rodents had improved survival and reduced incidence and/or severity of PhIP-induced neoplastic lesions compared to the PhIP-alone treated group. Rats eating tomato + broccoli exhibited a marked decrease in the number and size of cribiform prostatic intraepitheilial neoplasia/carcinoma in situ (cribiform PIN/CIS lesions and in the incidence of invasive intestinal adenocarcinomas and skin carcinomas. Although the apparent toxic effects of combined PhIP and tomato + broccoli need additional study, the results of this study support the hypothesis that a diet rich in tomato and broccoli can reduce or prevent dietary carcinogen-induced cancers.

  7. Dietary chemoprevention of PhIP induced carcinogenesis in male Fischer 344 rats with tomato and broccoli.

    Science.gov (United States)

    Canene-Adams, Kirstie; Sfanos, Karen S; Liang, Chung-Tiang; Yegnasubramanian, Srinivasan; Nelson, William G; Brayton, Cory; De Marzo, Angelo M

    2013-01-01

    The heterocyclic amine, 2-amino-1-methyl-6-phenylimidazo[4,5-B]pyridine (PhIP), found in meats cooked at high temperatures, has been implicated in epidemiological and rodent studies for causing breast, prostate, and colorectal cancers. A previous animal study using a xenograft model has shown that whole tomato and broccoli, when eaten in combination, exhibit a marked effect on tumor reduction compared to when eaten alone. Our aim was to determine if PhIP-induced carcinogenesis can be prevented by dietary consumption of whole tomato + broccoli powders. Male Fischer 344 rats (n = 45) were randomized into the following treatment groups: control (AIN93G diet), PhIP (200 ppm in AIN93G diet for the first 20 weeks of the study), or tomato + broccoli + PhIP (mixed in AIN93G diet at 10% each and fed with PhIP for 20 weeks, and then without PhIP for 32 weeks). Study animals were monitored for 52 weeks and were euthanized as necessary based on a set of criteria for health status and tumor burden. Although there appeared to be some hepatic and intestinal toxicity due to the combination of PhIP and tomato + broccoli, these rodents had improved survival and reduced incidence and/or severity of PhIP-induced neoplastic lesions compared to the PhIP-alone treated group. Rats eating tomato + broccoli exhibited a marked decrease in the number and size of cribiform prostatic intraepitheilial neoplasia/carcinoma in situ (cribiform PIN/CIS) lesions and in the incidence of invasive intestinal adenocarcinomas and skin carcinomas. Although the apparent toxic effects of combined PhIP and tomato + broccoli need additional study, the results of this study support the hypothesis that a diet rich in tomato and broccoli can reduce or prevent dietary carcinogen-induced cancers. PMID:24312188

  8. Detection of the Diversity of Cytoplasmic Male Sterility Sources in Broccoli (Brassica Oleracea var. Italica) Using Mitochondrial Markers.

    Science.gov (United States)

    Shu, Jinshuai; Liu, Yumei; Li, Zhansheng; Zhang, Lili; Fang, Zhiyuan; Yang, Limei; Zhuang, Mu; Zhang, Yangyong; Lv, Honghao

    2016-01-01

    Broccoli (Brassica oleracea var. italica) is an important commercial vegetable crop. As part of an efficient pollination system, cytoplasmic male sterility (CMS) has been widely used for broccoli hybrid production. Identifying the original sources of CMS in broccoli accessions has become an important part of broccoli breeding. In this study, the diversity of the CMS sources of 39 broccoli accessions, including 19 CMS lines and 20 hybrids, were analyzed using mitochondrial markers. All CMS accessions contained the ogu orf138-related DNA fragment and the key genes of nap CMS, pol CMS, and tour CMS were not detected. The 39 CMS accessions were divided into five groups using six orf138-related and two simple sequence repeat markers. We observed that ogu CMS R3 constituted 79.49% of the CMS sources. CMS6 and CMS26 were differentiated from the other accessions using a specific primer. CMS32 was distinguished from the other accessions based on a 78-nucleotide deletion at the same locus as the orf138-related sequence. When the coefficient was about 0.90, five CMS accessions (13CMS6, 13CMS23, 13CMS24, 13CMS37, and 13CMS39) exhibiting abnormal floral organs with poor seed setting were grouped together. The polymerase chain reaction amplification profiles for these five accessions differed from those of the other accessions. We identified eight useful molecular markers that can be used to detect CMS types during broccoli breeding. Our data also provide important information relevant to future studies on the possible origins and molecular mechanisms of CMS in broccoli. PMID:27446156

  9. EFFECTS OF PLANTING DATES AND VARIETIES ON GROWTH AND YIELD OF BROCCOLI DURING RAINY SEASON

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    Karistsapol Nooprom

    2013-01-01

    Full Text Available Broccoli (Brassica oleracea var. italica is one of the most popular vegetable crops in many countries of the world as its nutritional importance. It has the most exacting climatic and cultural requirements, which limit its commercial production to a few favored locations. In Thailand, broccoli has been expending in recent years and amount of increasing production. It is best planted in the North where the weather is cool or in the Northeast during winter season. The South has tropical climates and raining all year round, but farmers can produce broccoli during early summer by using the heat-tolerant varieties. Although the broccoli is highly priced and still wanted by consumers in the rainy season, the farmers might not choose to grow as the problems with low productivity. Thus, the selection of suitable planting dates and varieties is crucial for successful cultivation during rainy season in the South. The experiment was performed to evaluate the effects of planting dates and broccoli varieties on their growth and yield responses during rainy season. Four broccoli varieties: ‘Top Green’, ‘Green Queen’, ‘Yok Kheo’ and ‘Special’ were plated for six planting dates: July, August, September, October, November and December at Prince of Songkla University, Thailand, between July, 2011 and January, 2012. Split-plot in a randomized complete block design was used in this experiment with four replications. This study showed that four broccoli varieties had highly seedling survival rates of 70.31-99.22% when planting in July-December except ‘Special’ of 54.68% when planting in August. ‘Yok Kheo’ had plant diameter of 67.54-75.24 cm when planting in July and December, not significantly different from ‘Top Green’ planting in July, August and November of 69.15-71.24 cm and ‘Green Queen’ in July and September of 68.69-68.88 cm. The highest total yield was obtained by ‘Yok Kheo

  10. Assessing the anticancer compounds Se-methylselenocysteine and glucosinolates in Se-biofortified broccoli (brassica oleracea L. var. italica) sprouts and florets

    Science.gov (United States)

    Broccoli (Brassica oleracea L. var. italica) is a rich source of chemopreventive compounds. Here, we evaluated and compared the effect of selenium (Se) treatment on the accumulation of anticancer compound Se-methylselenocysteine (SeMSCys) and glucosinolates in broccoli sprouts and florets. Total Se ...

  11. Monolignol 4-O-methyltransferases and uses thereof

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Chang-Jun; Bhuiya, Mohammad-Wadud; Zhang, Kewei

    2014-11-18

    Modified (iso)eugenol 4-O-methyltransferase enzymes having novel capacity for methylation of monolignols and reduction of lignin polymerization in plant cell wall are disclosed. Sequences encoding the modified enzymes are disclosed.

  12. The inheritance of human erythrocyte catechol-O-methyltransferase activity

    International Nuclear Information System (INIS)

    Erythrocyte catechol-O-methyltransferase (COMT) activity was analyzed in 107 individuals from 23 families. The observed activities were consistent with an autosomal codominant inheritance of the gene coding for erythrocyte COMT activity. (author)

  13. Hypnotizability and Catechol-O-Methyltransferase (COMT) polymorphysms in Italians

    OpenAIRE

    Alessandro Gialluisi; Serena Barbuti; Giancarlo Carli; Enrica Laura Santarcangelo

    2014-01-01

    Higher brain dopamine content depending on lower activity of Catechol-O-Methyltransferase (COMT) in subjects with high hypnotisability scores (highs) has been considered responsible for their attentional characteristics. However, the results of the previous genetic studies on association between hypnotisability and the Catechol-O-Methyltransferase (COMT) single nucleotide polymorphism (SNP) rs4680 (Val158Met) were inconsistent. Here, we used a selective genotyping approach to re-evaluate the ...

  14. Effects of industrial pre-freezing processing and freezing handling on glucosinolates and antioxidant attributes in broccoli florets.

    Science.gov (United States)

    Cai, Congxi; Miao, Huiying; Qian, Hongmei; Yao, Leishuan; Wang, Bingliang; Wang, Qiaomei

    2016-11-01

    The effects of industrial pre-freezing processing and freezing handling on the contents of glucosinolates and antioxidants (vitamin C, polyphenols, carotenoid and chlorophyll), as well as the antioxidant capacity in broccoli (Brassica oleracea L. var. italica) florets were investigated in the present study. Our results showed that the glucosinolate accumulations were significantly decreased after pre-freezing processing, whereas elevated levels of phenols, carotenoids, chlorophyll, and also antioxidant capacity were observed in frozen broccoli florets. The contents of vitamin C remained constant during above mentioned processing. In conclusion, the current industrial freezing processing method is a good practice for the preservation of main antioxidant nutrients in broccoli florets, although some improvements in pre-freezing processing, such as steam blanching and ice-water cooling, are needed to attenuate the decrease in glucosinolate content. PMID:27211670

  15. Sulforaphane formation and bioaccessibility are more affected by steaming time than meal composition during in vitro digestion of broccoli.

    Science.gov (United States)

    Sarvan, I; Kramer, E; Bouwmeester, H; Dekker, M; Verkerk, R

    2017-01-01

    Broccoli is a rich source of the glucosinolate glucoraphanin (GR). After hydrolysis of GR by the endogenous enzyme myrosinase, sulforaphane (SF) or sulforaphane nitrile (SFN) are produced, depending on environmental conditions. How the conversion of GR and bioaccessibility of released breakdown products are affected by steaming (raw, 1min, 2min and 3min steamed) and meal composition (protein or lipid addition) was studied with an in vitro digestion model (mouth, stomach, intestine, but not colonic digestion). The main formation of SF and SFN occurred during in vitro chewing. The contents of GR, SF and SFN did not change after further digestion, as the irreversible inactivated myrosinase under gastric conditions caused no further GR hydrolysis. SF concentrations were up to 10 times higher in raw and 1min steamed broccoli samples after digestion compared to longer-steamed broccoli. Protein or lipid addition had no influence on the formation and bioaccessibility of SF or SFN. PMID:27507513

  16. Effect of Planting Time on Incidence of Bacterial Disease and Yield of Broccoli during Dry Season in Southern Thailand

    OpenAIRE

    Karistsapol Nooprom; Quanchit Santipracha; Sompong Te-chato

    2013-01-01

    The trial was aimed to select suitable planting times when broccoli varieties could resist well to soft rot and black rot diseases during dry season was studies at Prince of Songkla University, Songkhla, Thailand, from January to June, 2011. Split-plot in randomized complete block design was used in this experiment with four replications. The results showed that the lowest incidence of soft rot disease was observed in all broccoli varieties when they were planted in January (4.19-7.18%). Afte...

  17. Methyl Jasmonate and 1-Methylcyclopropene Treatment Effects on Quinone Reductase Inducing Activity and Post-Harvest Quality of Broccoli

    OpenAIRE

    Ku, Kang Mo; Choi, Jeong Hee; Kim, Hyoung Seok; Kushad, Mosbah M.; Jeffery, Elizabeth H.; Juvik, John A.

    2013-01-01

    Effect of pre-harvest methyl jasmonate (MeJA) and post-harvest 1-methylcyclopropene (1-MCP) treatments on broccoli floret glucosinolate (GS) concentrations and quinone reductase (QR, an in vitro anti-cancer biomarker) inducing activity were evaluated two days prior to harvest, at harvest and at 10, 20, and 30 days of post-harvest storage at 4 °C. MeJA treatments four days prior to harvest of broccoli heads was observed to significantly increase floret ethylene biosynthesis resulting in chloro...

  18. IDENTIFICATION OF ANTAGONISTS OF Xanthomonas campestris ISOLATED FROM RHIZOSPHERE ZONE OF BROCCOLI FARM AT KEMBANG MERTA VILLAGE, TABANAN, BALI

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    Nadya Treesna Wulansari

    2015-03-01

    Full Text Available The main objectives of this research were to isolate and identify antagonists of Xanthomonas campestris from rhizosphere zone of broccoli plants. Soil samples were collected from broccoli farm located at Kembang Merta village, Tabanan, Bali. Isolation and identification of the antagonists were conducted at the Laboratory of Microbiology, Udayana University. Two fungal (Trichoderma harzianum and Trichoderma viride and two bacterial (Bacillus sp. and Pseudomonas sp. antagonists potentially to be developed as biocontrol agents of Xanthomonas campestris were successfully identified in this research

  19. The Impact of Broccoli II & Tomato II on European patents in conventional breeding, GMO’s and Synthetic Biology:

    DEFF Research Database (Denmark)

    Minssen, Timo; Nordberg, Ana

    2015-01-01

    On 25 March 2015, the Enlarged Board of Appeal of the European Patent Office (EBA) finally delivered its’ much awaited decisions on the consolidated referrals G2/12 (“Tomato II”) and G2/13 (“Broccoli II”). The EBA affirmed that products, namely plants or parts thereof, obtained by essentially...... (GMOs) and Synthetic Biology remains a major obstacle to the industry, research and innovation in an increasingly significant area of science. This paper has also been published as a working paper on SSRN, see: Minssen, Timo and Nordberg, A., The Impact of Broccoli II & Tomatoes II on European Patents...

  20. Optimisation of the Method for the Quantitative Determination of Sulforaphane in Broccoli

    International Nuclear Information System (INIS)

    Consumption of vegetables, especially crucifers, reduces the risk of developing cancer. Sulforaphane [l-isothiocyanato-4- (methylsulfinyl)-butane], a compound with the ability to inhibit carcinogenesis, is one of the degradation products of glucosinolates in cruciferous vegetables. Among available extraction methods, autolysis at room temperature is the most effective for Sulforaphane extraction (relatively higher purity and better yield). The research work undertaken at Federal Research Centre for Nutrition, Institute of Biology and Chemistry, Karlsruhe, Germany was isolation of Sulforaphane based on cruciferous vegetables like Broccoli (Brassica oleracea L. Cv. italica) employing autolysis - the yield being higher. The extracted Sulforaphane compound's purity and yield were accordingly examined with gas chromatography. (author)

  1. Purification and properties of thioether methyltransferase

    International Nuclear Information System (INIS)

    A method to assay activity was developed which measures acceptance of methyl groups from [methyl-3H]-S-adenosylmethionine by dimethyl selenide. The product, [3H]trimethylselenonium ion, is separated by HPLC and quantitated by scintillation counting. Thioether methyltransferase from mouse liver and lung resides primarily in the cytosol. In terms of specific activity the enzyme is most active in the lung and liver. Purification from lung cytosol requires a three-step process of DEAE and gel filtration column chromatographies followed by chromatofocusing. SDS-Polyacrylamide gel electrophoresis shows a single homogeneous band with a molecular mass of 28,000 daltons. Vmax and Km values for dimethyl selenide as a substrate are 15. 7 pmol/min and 0.44 μM, respectively. Our studies have also shown that this purified enzyme is capable of methylating a wide range of compounds. To further test the enzyme's role in detoxification, in vivo studies were performed by injecting mice with substrate and [methyl-3H]methionine and analyzing tissue extracts and urine for [methyl-3H]sulfonium

  2. Isolation of DNA methyltransferase from plants

    International Nuclear Information System (INIS)

    DNA methyltransferases (DMT) were isolated from nuclei of cauliflower, soybean, and pea by extraction with 0.35 M NaCl. Assays were performed on hemimethylated Micrococcus luteus DNA or on M. luteus DNA to test for maintenance or de novo methylase activity, respectively. Fully methylated DNA was used as a substrate to determine background levels of methylation. Based on these tests, yields of maintenance DMT activity in the crude extract from pea hypocotyl, soybean hypocotyl, and cauliflower inflorescence were 2.8, 0.9, and 1.6 units per g wet tissue (one unit equals 1 pmol of methyl from [3H]AdoMet incorporated into acid precipitable material per h at 300). Two peaks of DMT activity were detected in the soybean nuclear extract following phosphocellulose chromatography. One eluted at 0.4 M and the other at 0.8 M KCl. With both fractions maintenance activity was approximately 2 times that of the de novo activity. Using gel filtration the DMT eluted at 220,000 Daltons. The optimal pH for activity was between 6.5 and 7.0, and the optimal temperature was 300

  3. Incidence of Bacterial Disease and Yield of Broccoli as Influenced by Different Rain Protectors and Varieties during the Rainy Season in Southern Thailand

    Directory of Open Access Journals (Sweden)

    Karistsapol Nooprom

    2014-04-01

    Full Text Available This study is mainly focused on evaluating the effects of different rain protectors and broccoli varieties to find out whether rain protector and variety is suitable or not for broccoli production during rainy season. Broccoli was experimented at Prince of Songkla University, Hat Yai, Songkhla, southern Thailand. Study revealed that broccoli growing under plastic sheet and green shade net had lower incidence of soft rot disease (1.62 and 3.75%, respectively than those grown in open field (13.33% while their growing under plastic sheet (1.50% had lower incidence of black rot disease than those grown under green shade net and open field (18.75 and 32.88%, respectively. All broccoli varieties were found to be statistically different in their response towards soft rot and black rot diseases. However, the Top Green had the highest diseases incidence (8.33 and 21.08%, respectively while the Yok Kheo had the lowest diseases incidence (4.62 and 0.00%, respectively. The highest total yield was obtained from the Yok Kheo when those grown under plastic sheet (13.48 t/ha while the Top Green had lowest yield when those grown in open field (3.94 t/ha. Therefore, the most suitable method for broccoli production during rainy season in southern Thailand was to grow under plastic sheet and green shade net by using the three varieties of broccoli.

  4. Phenotypic and genotypic characterization of Xanthomonas campestris strains isolated from cabbage, kale and broccoli

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    Popović Tatjana

    2013-01-01

    Full Text Available Thirty-six strains of Xanthomonas campestris pv. campestris (Xcc isolated from cabbage, kale and broccoli were identified according to their pathogenicity, phenotypic and genotypic characterization. Pathogenicity was confirmed by the injection method with a hypodermic syringe into the mesophilic tissue of cabbage leaves. All strains were Gramnegative, aerobic, catalase-positive, oxidase-negative, grew at 35°C, produced levan, H2S and indole, did not reduce nitrate, hydrolyzed Tween 80, starch, gelatin and esculin and did not show tolerance to 0.1 and 0.02% TTC. The strains produced acid from d-arabinose, arginine, dulcitol, galactose, d-glucose, maltose, mannose, sorbitol, sucrose and xylose. The genetic characterization was based on the sequence analyses of 16S rDNA and ERIC and BOX PCR. Strains of different pathovars were also used to compare PCR resulting patterns. BOX-PCR of the strains from kale and broccoli, obtained using (GTG5 primer, yielded patterns with a high similarity level to pathovar reference strain Xcc. The strains from cabbage yielded BOX and ERIC product patterns, distinguishing them from the other tested strains and reference strains. 16S rDNA of the representative strains was closely related to Xcc strain ATCC 33913. ERIC PCR and BOX using (GTG5 primer generated different Xcc patterns and were effective in distinguishing strains from different plant hosts. [Projekat Ministarstva nauke Republike Srbije, br. III43010 i br. III46007

  5. Optimizing elicitation and seed priming to enrich broccoli and radish sprouts in glucosinolates.

    Science.gov (United States)

    Baenas, Nieves; Villaño, Debora; García-Viguera, Cristina; Moreno, Diego A

    2016-08-01

    Elicitation is a cheaper and socially acceptable tool for improving plant food functionality. Our objective was to optimize the treatment doses of the elicitors: methyl jasmonate (MeJA), jasmonic acid (JA) and DL-methionine (MET), in order to find a successful and feasible treatment to produce broccoli and radish sprouts with enhanced levels of health-promoting glucosinolates. Also a priming of seeds as a novel strategy to trigger the glucosinolates content was carried out with water (control), MeJA (250μM), JA (250μM) and MET (10mM) before the elicitor exogenous treatment. The results showed that almost all treatments could enhance effectively the total glucosinolates content in the sprouts, achieving the most significant increases from 34% to 100% of increase in broccoli and from 45% to 118% of increase in radish sprouts after MeJA priming and treatments. Consequently, our work demonstrates the feasibility of using elicitors, such as plant stress hormones, by priming and exogenously, as a way of increase the phytochemical profile of these sprouts to enhance their consumption in the diet. PMID:26988507

  6. Pectobacterium carotovorum subsp. carotovorum – the causal agent of broccoli soft rot in Serbia

    Directory of Open Access Journals (Sweden)

    Katarina Gašić

    2014-12-01

    Full Text Available Soft rot symptoms were observed on broccoli plants in several commercial fields in the western part of Serbia. Six strains of bacteria were isolated from diseased tissues and identified as Pectobacterium carotovorum subsp. carotovorum using conventional bacteriological and molecular methods. All strains were non-fluorescent, gram-negative, facultative anaerobes, oxidase-negative and catalase-positive, causing soft rot on potato and carrot slices and did not induce hypersensitive reaction on tobacco leaves. They grew in 5% NaCl and at 37°C, did not produce acid from α-methyl glucoside, sorbitol and maltose, nor reducing substances from sucrose, but utilized lactose and trehalose, and did not produce indole or lecithinase. The investigated strains showed characteristic growth on Logan’s medium and did not produce blue pigmented indigoidine on GYCA medium nor “fried egg” colonies on PDA. The identity of strains was confirmed by ITS-PCR and ITS-RFLP analyses and by sequence analysis of the 16S rRNA gene. In a pathogenicity assay, all strains caused tissue discoloration and soft rot development on inoculated broccoli head tissue fragments.

  7. Stingless bees damage broccoli inflorescences when collecting fibers for nest building

    Directory of Open Access Journals (Sweden)

    Adriano Jorge Nunes dos Santos

    2012-01-01

    Full Text Available The stingless bee Trigona spinipes (Fabricius, 1793 (Hymenoptera: Apidae is an important pollinator for various crops, but constitutes an occasional pest of other plant species since it causes injury to leaves, stems, flowers and fruits while collecting nest materials. The aim of the present study was to determine the damage caused by T. spinipes to a broccoli (Brassica oleracea L. var. italica, Brassicaceae growing on an organic farm. A significant number of plants (72.5 % presented damaged inflorescences, while 39% of all of the inflorescences suffered some degree of injury. The activities of T. spinipes caused scarifications on the stems of the inflorescences, and these typically evolved to epidermal cicatrices up to 10 mm wide. In some cases, the lesions were sufficiently deep to cause partial destruction of the vascular tissues, and this lead to thinner (< 5 mm diameter floral stems that may collapse. To the best of our knowledge, this is the first report concerning the attack of broccoli plants by T. spinipes. The results obtained should serve to highlight the possibility that stingless bees could be responsible for direct and/or indirect damage to vegetable crops, and to stimulate the development of control strategies for these incidental pests.

  8. Biotechnological applications in in vitro plant regeneration studies of broccoli (Brassica oleracea L. var. italica), an important vegetable crop.

    Science.gov (United States)

    Kumar, Pankaj; Srivastava, Dinesh Kumar

    2016-04-01

    Biotechnology holds promise for genetic improvement of important vegetable crops. Broccoli (Brassica oleracea L. var. italica) is an important vegetable crop of the family Brassicaceae. However, various biotic and abiotic stresses cause enormous crop yield losses during commercial cultivation of broccoli. Establishment of a reliable, reproducible and efficient in vitro plant regeneration system with cell and tissue culture is a vital prerequisite for biotechnological application of crop improvement programme. An in vitro plant regeneration technique refers to culturing, cell division, cell multiplication, de-differentiation and differentiation of cells, protoplasts, tissues and organs on defined liquid/solid medium under aseptic and controlled environment. Recent progress in the field of plant tissue culture has made this area one of the most dynamic and promising in experimental biology. There are many published reports on in vitro plant regeneration studies in broccoli including direct organogenesis, indirect organogenesis and somatic embryogenesis. This review summarizes those plant regeneration studies in broccoli that could be helpful in drawing the attention of the researchers and scientists to work on it to produce healthy, biotic and abiotic stress resistant plant material and to carry out genetic transformation studies for the production of transgenic plants. PMID:26721234

  9. Salinity’s influence on boron toxicity in broccoli: II. Impacts on boron uptake, uptake mechanisms and tissue ion relations.

    Science.gov (United States)

    Limited research has been conducted on the interactive effects of salinity and boron stresses on plants despite their common occurrence in natural systems. The purpose of this research was to determine and quantify the interactive effects of salinity, salt composition and boron on broccoli (Brassica...

  10. Discriminating between cultivars and treatments of broccoli using mass spectral fingerprinting and analysis of variance-principal component analysis

    Science.gov (United States)

    Metabolite fingerprints, obtained with direct injection mass spectrometry (MS) with both positive and negative ionization, were used with analysis of variance-principal components analysis (ANOVA-PCA) to discriminate between cultivars and growing treatments of broccoli. The sample set consisted of ...

  11. Alterations in Chlorophyll a Fluorescence and Pigments Concentration in the Leaves of Cauliflower and Broccoli Transplants Subjected to Chilling

    Directory of Open Access Journals (Sweden)

    Renata WOJCIECHOWSKA

    2016-06-01

    Full Text Available Chlorophyll a fluorescence parameters and photosynthetic pigments content in leaves of broccoli (Brassica oleracea L. var. italica cv. ‘Monaco’ F1 and cauliflower (Brassica oleracea L. var. botrytis cv. ‘Bruce’ F1 transplants were analyzed to investigate the effects of chilling temperatures (6 and 10 °C and the exposure length (1 or 2 weeks on the photosynthetic apparatus condition in 3-year experiment. Data were compared to control plants exposed to 14 °C for 1 and 2 weeks. The lowest values of maximum quantum efficiency of photosystem II (Fv/Fm and the performance index (PI were observed in cauliflower chilled at 6 °C for 2 weeks. This was not accompanied by any substantial reduction in chlorophylls concentration, however, significant correlations between Fv/Fm or PI and chlorophylls content in cauliflower leaves were found. There was no negative effects of chilling on photosynthetic activity and chlorophyll content in broccoli leaves, the amount of carotenoids significantly increased in the broccoli chilled for 2 weeks in 6 °C. The chlorophyll a fluorescence indices and accelerated carotenoids synthesis showed that broccoli had less vulnerable photosynthetic apparatus to stress-induced temperatures and more efficient protection mechanisms in comparison to cauliflower transplants.

  12. Pure phosphine fumigation treatment at low temperature for postharvest control of western flower thrips on lettuce, broccoli, asparagus, and strawberries

    Science.gov (United States)

    U.S. exported lettuce, broccoli, asparagus, and strawberries often harbor western flower thrips (Frankliniella occidentalis), a quarantined pest in Taiwan, and therefore require quarantine treatment. Pure phosphine fumigation at a low temperature of 2°C was studied as an alternative fumigant to meth...

  13. Virginia Cooperative Extension evaluates broccoli varieties for $4.9 million study on promising East Coast industry

    OpenAIRE

    Greiner, Lori A.

    2010-01-01

    When it comes to buying local, the cornucopia of East Coast produce runneth over with apples, peaches, pumpkins, and potatoes. Broccoli isn't a major part of the bounty just yet, but Virginia Cooperative Extension agent Wythe Morris hopes to change that.

  14. Sulforaphane formation and bioaccessibility are more affected by steaming time than meal composition during in vitro digestion of broccoli

    NARCIS (Netherlands)

    Sarvan-Kruse, Irmela; Kramer, E.; Bouwmeester, H.; Dekker, M.; Verkerk, R.

    2017-01-01

    Broccoli is a rich source of the glucosinolate glucoraphanin (GR). After hydrolysis of GR by the endogenous enzyme myrosinase, sulforaphane (SF) or sulforaphane nitrile (SFN) are produced, depending on environmental conditions. How the conversion of GR and bioaccessibility of released breakdown p

  15. Variation in Broccoli Cultivar Phytochemical Content under Organic and Conventional Management Systems: Implications in Breeding for Nutrition

    NARCIS (Netherlands)

    Renaud, E.N.C.; Lammerts Van Bueren, E.; Myers, J.R.; Caldas Paulo, M.J.; Eeuwijk, van F.A.; Zhu, N.; Juvik, J.A.

    2014-01-01

    Organic agriculture requires cultivars that can adapt to organic crop management systems without the use of synthetic pesticides as well as genotypes with improved nutritional value. The aim of this study encompassing 16 experiments was to compare 23 broccoli cultivars for the content of phytochemic

  16. DNA Methyltransferases Inhibitors from Natural Sources.

    Science.gov (United States)

    Zwergel, Clemens; Valente, Sergio; Mai, Antonello

    2016-01-01

    DNA methyltransferases (DNMTs) catalyze the methylation at cytosine-C5 mainly in a CpG dinucleotide context. Although DNA methylation is essential for fundamental processes like embryonic development or differentiation, aberrant expression and/or activities of DNMTs are involved in several pathologies, from neurodegeneration to cancer. DNMTs inhibition can arrest tumor growth, cells invasiveness and induce differentiation, whereas their increased expression is shown in numerous cancer types. Moreover, hypermethylated promoters of tumor suppressor genes lead to their silencing. Hence, the use of specific inhibitors of DNMT might reactivate those genes and stop or even reverse the aberrant cell processes. To date, the only approved DNMTs inhibitors for therapy belong to the nucleoside-based family of drugs, but they display relevant side effects as well as high chemical instability. Thus, there is a keen interest actually exists to develop novel, potent and safe inhibitors possessing a nonnucleoside structure. Increasing literature evidence is highlighting that natural sources could help the researchers to achieve this goal. Indeed, several polyphenols, flavonoids, antraquinones, and others are described able to inhibit DNMTs activity and/or expression, thus decreasing the methylation/silencing of different genes involved in tumorigenesis. These events can lead to re-expression of such genes and to cell death in diverse cancer cell lines. Epigallocatechin-3-gallate (1) and laccaic acid A (11) resulted the most effective DNMT1 inhibitors with submicromolar IC50 values, acting as competitive inhibitors. Compound 1 and 11 both displayed gene demethylation and re-activation in several cancers. However, all of the natural compounds described in this review showed important results, from gene reactivation to cell growth inhibition. Moreover, some of them displayed interesting activity even in rodent cancer models and very recently entered clinical trials. PMID:26303417

  17. Multiple lysine methylation of PCAF by Set9 methyltransferase

    International Nuclear Information System (INIS)

    The molecular functions of several non-histone proteins are regulated through lysine modification by histone methyltransferases. The p300/CBP-associated factor (PCAF) is an acetyltransferase that has been implicated in many cellular processes. Here, we report that PCAF is a novel substrate of Set9 methyltransferase. In vitro mapping experiments revealed six lysine residues could be methylated by Set9. A comparison of amino acid sequences of target sites revealed the novel consensus motif which differs from previously identified Set9-consensus sequence. Further methyltransferase assays focusing on the six lysine residues showed that K78 and K89 are preferentially methylated in full-length PCAF in vitro. Using specific antibodies recognizing mono-methylated K89, in vivo PCAF methylation and its nuclear localization were demonstrated. Our data may lead to a new insight into PCAF functions and provide additional information to identify unknown targets of Set9.

  18. NPK, protein content and yield of broccoli as affected by gamma rays seeds irradiation and phosphorus fertilizer rates

    International Nuclear Information System (INIS)

    Two field experiments were carried out during 1999/2000 and 2000/2001 winter growing seasons at the experimental farm of Nuclear Research Center, Atomic Energy Authority, Inshas, Egypt. The experiments were conducted to study the effect of pre sowing-seeds irradiation with different doses of gamma rays (0, 2, 3 and 4 Gy) and different phosphorus fertilizer application rates, 0, 30, 60 and 90 k P2O5 /fed) on NPK content of leaves and spear, and protein content in spears at maturity, spear diameter, main spear fresh and dry weight per plant, total spear fresh weight per plant and total spear yield. In general, exposing broccoli seeds to different gamma ray doses up to 4 Gy prior to sowing increased the above mentioned parameters with different magnitudes comparing with the non-irradiated control plants. The highest percentage of increase was obtained by exposing broccoli seeds to 3 Gy. There were non-significant differences between 3 and 4 Gy treatments during the two growing seasons. With respect to the effect of phosphorus fertilizer application rates on the studied parameters, increasing phosphorus application rates up to 90 kg P2O5/fed increased the above mentioned parameters. The highest percentage of increase was obtained by applying 90 kg P2O5/fed. The interaction, gamma ray and P level showed phosphorus there were significant differences in main spear fresh and dry weight per plant, total spear yield and spear diameter in first season. The highest value was obtained by 3 Gy and 90 kg P2O5/fed. Also there were significant effects on NPK content in broccoli leaves at 90 days after transplanting (DAT) except P in second season and nonsignificant values of broccoli spear at harvest except N, K in first season. The highest protein content of broccoli spears at harvest was obtained with 2 Gy and 30 kg P25/fed

  19. Plants as Biofactories: Postharvest Stress-Induced Accumulation of Phenolic Compounds and Glucosinolates in Broccoli Subjected to Wounding Stress and Exogenous Phytohormones

    OpenAIRE

    Villarreal-García, Daniel; Nair, Vimal; Cisneros-Zevallos, Luis; Daniel A. Jacobo-Velázquez

    2016-01-01

    Broccoli contains high levels of bioactive molecules and is considered a functional food. In this study, postharvest treatments to enhance the concentration of glucosinolates and phenolic compounds were evaluated. Broccoli whole heads were wounded to obtain florets and wounded florets (florets cut into four even pieces) and stored for 24 h at 20 °C with or without exogenous ethylene (ET, 1000 ppm) or methyl jasmonate (MeJA, 250 ppm). Whole heads were used as a control for wounding treatments....

  20. Chromosomal replication incompatibility in Dam methyltransferase deficient Escherichia coli cells

    DEFF Research Database (Denmark)

    Freiesleben, Ulrik Von

    1996-01-01

    Dam methyltransferase deficient Escherichia coli cells containing minichromosomes were constructed. Free plasmid DNA could not be detected in these cells and the minichromosomes were found to be integrated in multiple copies in the origin of replication (oriC) region of the host chromosome. The...

  1. Label-free electrochemical detection of human methyltransferase from tumors.

    Science.gov (United States)

    Furst, Ariel L; Muren, Natalie B; Hill, Michael G; Barton, Jacqueline K

    2014-10-21

    The role of abnormal DNA methyltransferase activity in the development and progression of cancer is an essential and rapidly growing area of research, both for improved diagnosis and treatment. However, current technologies for the assessment of methyltransferase activity, particularly from crude tumor samples, limit this work because they rely on radioactivity or fluorescence and require bulky instrumentation. Here, we report an electrochemical platform that overcomes these limitations for the label-free detection of human DNA(cytosine-5)-methyltransferase1 (DNMT1) methyltransferase activity, enabling measurements from crude cultured colorectal cancer cell lysates (HCT116) and biopsied tumor tissues. Our multiplexed detection system involving patterning and detection from a secondary electrode array combines low-density DNA monolayer patterning and electrocatalytically amplified DNA charge transport chemistry to measure selectively and sensitively DNMT1 activity within these complex and congested cellular samples. Based on differences in DNMT1 activity measured with this assay, we distinguish colorectal tumor tissue from healthy adjacent tissue, illustrating the effectiveness of this two-electrode platform for clinical applications. PMID:25288757

  2. Changes in SeMSC, Glucosinolates and Sulforaphane Levels, and in Proteome Profile in Broccoli (Brassica oleracea var. Italica Fertilized with Sodium Selenate

    Directory of Open Access Journals (Sweden)

    Alejandra Moenne

    2013-05-01

    Full Text Available The aim of this work was to analyze the effect of sodium selenate fortification on the content of selenomethyl selenocysteine (SeMSC, total glucosinolates and sulforaphane, as well as the changes in protein profile of the inflorescences of broccoli (Brassica oleracea var. Italica. Two experimental groups were considered: plants treated with 100 mmol/L sodium selenate (final concentration in the pot and control plants treated with water. Fortification began 2 weeks after transplantation and was repeated once a week during 10 weeks. Broccoli florets were harvested when they reached appropriate size. SeMSC content in broccoli florets increased significantly with sodium selenate fortification; but total glucosinolates and sulforaphane content as well as myrosinase activity were not affected. The protein profile of broccoli florets changed due to fortification with sodium selenate. Some proteins involved in general stress-responses were up-regulated, whereas down-regulated proteins were identified as proteins involved in protection against pathogens. This is the first attempt to evaluate the physiological effect of fortification with sodium selenate on broccoli at protein level. The results of this work will contribute to better understanding the metabolic processes related with selenium uptake and accumulation in broccoli.

  3. Effect of blanching temperature and dipping time on drying time of broccoli.

    Science.gov (United States)

    Doymaz, Ibrahim

    2014-03-01

    Drying characteristics of broccoli was experimentally studied in cabinet-type convective dryer. Experiments were conducted using constant air velocity of 2 m/s and air temperature of 60 . Drying rate increased with increasing blanching temperature and thus reduced the drying time. The experimental drying data were used to fit twelve drying models and drying rate constants and coefficients of the models were determined by non-linear regression analysis. Estimations by the Midilli et al. model were in good agreement with the experimental data obtained. Fick's second law was used to calculate the effective moisture diffusivity, which varied from 1.987 to 3.577 × 10(-8) m(2)/s for the given blanching temperature range. The rehydration ratio was observed to increase with increasing blanching temperature. PMID:23744113

  4. 秋花椰菜引种试验%Trial of introducing autumn broccoli

    Institute of Scientific and Technical Information of China (English)

    蔡文注; 叶明鑫; 陈艺婷; 颜慧莹; 陈龙杰

    2012-01-01

    Trial test of 13 introduced broccoli varieties has been carried out in Xiang' an district of Xiamen city in autumn of 2011. Two qualified and resistant varieties SWONSEA and SUPER FORMEN were screened by comparison of plant chacteristicrs, resistance, corm pattern, single corm weight, yield and quality.%2011年秋季在厦门市翔安区对引入的13个花椰菜品种进行试种,通过植株性状、抗逆性、球型、单球重、产量及品质的比较,初步筛选出优质、抗性好的秋花椰菜品种SWONSEA和SUPERFORMEN。

  5. Structure and Function of Flavivirus NS5 Methyltransferase

    Energy Technology Data Exchange (ETDEWEB)

    Zhou,Y.; Ray, D.; Zhao, Y.; Dong, H.; Ren, S.; Li, Z.; Guo, Y.; Bernard, K.; Shi, P.; Li, H.

    2007-01-01

    The plus-strand RNA genome of flavivirus contains a 5' terminal cap 1 structure (m{sup 7}GpppAmG). The flaviviruses encode one methyltransferase, located at the N-terminal portion of the NS5 protein, to catalyze both guanine N-7 and ribose 2'-OH methylations during viral cap formation. Representative flavivirus methyltransferases from dengue, yellow fever, and West Nile virus (WNV) sequentially generate GpppA {yields} m{sup 7}GpppA {yields} m{sup 7}GpppAm. The 2'-O methylation can be uncoupled from the N-7 methylation, since m{sup 7}GpppA-RNA can be readily methylated to m{sup 7}GpppAm-RNA. Despite exhibiting two distinct methylation activities, the crystal structure of WNV methyltransferase at 2.8 {angstrom} resolution showed a single binding site for S-adenosyl-L-methionine (SAM), the methyl donor. Therefore, substrate GpppA-RNA should be repositioned to accept the N-7 and 2'-O methyl groups from SAM during the sequential reactions. Electrostatic analysis of the WNV methyltransferase structure showed that, adjacent to the SAM-binding pocket, is a highly positively charged surface that could serve as an RNA binding site during cap methylations. Biochemical and mutagenesis analyses show that the N-7 and 2'-O cap methylations require distinct buffer conditions and different side chains within the K{sub 61}-D{sub 146}-K{sub 182}-E{sub 218} motif, suggesting that the two reactions use different mechanisms. In the context of complete virus, defects in both methylations are lethal to WNV; however, viruses defective solely in 2'-O methylation are attenuated and can protect mice from later wild-type WNV challenge. The results demonstrate that the N-7 methylation activity is essential for the WNV life cycle and, thus, methyltransferase represents a novel target for flavivirus therapy.

  6. Engineering Monolignol 4-O-Methyltransferases to Modulate Lignin Biosynthesis

    Energy Technology Data Exchange (ETDEWEB)

    Bhuiya, M.W.; Liu, C.

    2010-01-01

    Lignin is a complex polymer derived from the oxidative coupling of three classical monolignols. Lignin precursors are methylated exclusively at the meta-positions (i.e. 3/5-OH) of their phenyl rings by native O-methyltransferases, and are precluded from substitution of the para-hydroxyl (4-OH) position. Ostensibly, the para-hydroxyls of phenolics are critically important for oxidative coupling of phenoxy radicals to form polymers. Therefore, creating a 4-O-methyltransferase to substitute the para-hydroxyl of monolignols might well interfere with the synthesis of lignin. The phylogeny of plant phenolic O-methyltransferases points to the existence of a batch of evolutionarily 'plastic' amino acid residues. Following one amino acid at a time path of directed evolution, and using the strategy of structure-based iterative site-saturation mutagenesis, we created a novel monolignol 4-O-methyltransferase from the enzyme responsible for methylating phenylpropenes. We show that two plastic residues in the active site of the parental enzyme are vital in dominating substrate discrimination. Mutations at either one of these separate the evolutionarily tightly linked properties of substrate specificity and regioselective methylation of native O-methyltransferase, thereby conferring the ability for para-methylation of the lignin monomeric precursors, primarily monolignols. Beneficial mutations at both sites have an additive effect. By further optimizing enzyme activity, we generated a triple mutant variant that may structurally constitute a novel phenolic substrate binding pocket, leading to its high binding affinity and catalytic efficiency on monolignols. The 4-O-methoxylation of monolignol efficiently impairs oxidative radical coupling in vitro, highlighting the potential for applying this novel enzyme in managing lignin polymerization in planta.

  7. Ozone dose-response relationships for spring oilseed rape and broccoli

    Science.gov (United States)

    De Bock, Maarten; Op de Beeck, Maarten; De Temmerman, Ludwig; Guisez, Yves; Ceulemans, Reinhart; Vandermeiren, Karine

    2011-03-01

    Tropospheric ozone is an important air pollutant with known detrimental effects for several crops. Ozone effects on seed yield, oil percentage, oil yield and 1000 seed weight were examined for spring oilseed rape ( Brassica napus cv. Ability). For broccoli ( Brassica oleracea L. cv. Italica cv. Monaco) the effects on fresh marketable weight and total dry weight were studied. Current ozone levels were compared with an increase of 20 and 40 ppb during 8 h per day, over the entire growing season. Oilseed rape seed yield was negatively correlated with ozone dose indices calculated from emergence until harvest. This resulted in an R2 of 0.24 and 0.26 ( p rape. The reduction of oilseed rape yield showed the highest correlation with the ozone uptake during the vegetative growth stage: when only the first 47 days after emergence were used to calculate POD 6, R2 values increased up to 0.476 or even 0.545 when the first 23 days were excluded. The highest ozone treatments, corresponding to the future ambient level by 2100 (IPCC, Meehl et al., 2007), led to a reduction of approximately 30% in oilseed rape seed yield in comparison to the current ozone concentrations. Oil percentage was also significantly reduced in response to ozone ( p < 0.001). As a consequence oil yield was even more severely affected by elevated ozone exposure compared to seed yield: critical levels for oil yield dropped to 3.2 ppm h and 3.9 mmol m -2. For broccoli the applied ozone doses had no effect on yield.

  8. Re-translocation of boron in broccoli and lupin during early reproductive growth

    International Nuclear Information System (INIS)

    The objective of the present study was to determine if boron (B) re-translocation depends on plant-B status and external-B supply. The stable 10B isotope was supplied to the root system of broccoli (Brassica oleracea var. italica Plenck cv. Commander) and lupin (Lupinus albus L. cv. Ultra) plants to provide a quantitative picture of B distribution during early reproductive development. Regardless of the B regime (i.e. continuous supply with luxury, sufficient or deficient B; transfer at influorescence emergence from either a luxury- or sufficient-B supply to a deficient one) and whether 10B was acquired before or during influorescence development, a significant proportion of the B recovered in broccoli florest and lupin fruit was 10B enriched. B acquired during inflorescence development was an important source of B for reproductive structures, but the relative importance of B acquired before and after inflorescence emergence appeared to be species dependent. The occurrence of B re-translocation was not dependent upon the induction of B deficiency. The concentrations of B in phloem exsudates (0.38 to 0.03 mM) were 4- to 23-fold those in xylem sap, and more similar to the concentrations in the reproductive structures (0.86 to 0.07 mM) than those in source leaves (2.4 to 0.19 mM). The decreasing acropetal gradient of tissue-B concentrations with luxury-B supply declined dramatically or was reversed in plants grown with sufficient or deficient B. The data are consistent with B being a phloem-mobile element, and suggest that newly acquired B is particularly important during the early reproductive growth of plants. (au) 36 refs

  9. Vegetative and reproductive plasticity of broccoli at three levels of incident photosynthetically active radiation

    International Nuclear Information System (INIS)

    To study the effects of shading on the growth, development, dry matter partitioning, and plant architecture of broccoli, ‘Legacy’ hybrid plants were grown in pots in a greenhouse under black shading meshes to generate different levels of photosynthetically active radiation (PAR). The average incident PAR was 23 mol PPF m–2 d–1 under control conditions, 15.2 under a 35% mesh, and 6.7 under a 70% mesh. The air temperature was within the range 15-22°C. As shading increased so did the duration of the growth cycle and the leaf area (LA). Shading did not affect the number of leaves, although the upper ones were more erect. The stem length and the accumulated intercepted PAR (IPAR) were negatively related. Inflorescence diameter and commercial fresh weight decreased only with the 70% mesh. Shading did not affect stem dry weight (DW) but altered dry matter allocation in the root and spear. The DW of the leaves maintained an average 45.7% of the total DW despite the greater LA developed under shade. The net assimilation rate diminished with the reduction of IPAR, and the LA increased; the plant relative growth rate was therefore practically constant. With increased shading, the leaves and the stem became the dominant photosynthate sinks. The commercial fresh weight achieved with 15.2 mol photosynthetic photon flux (PPF) m–2 d–1 was equivalent to that obtained with 23 mol PPF m–2 d–1, but the cycle was extended for 4.5 days. With 6.7 mol PPF m–2 d–1, yield diminished because of the lower DW produced in a cycle extended by 15 days, and because more dry matter was allocated to the stem than to the spear. Based on these results, broccoli could be considered a shade-tolerant plant. (author)

  10. Molecular identification of carnosine N-methyltransferase as chicken histamine N-methyltransferase-like protein (hnmt-like.

    Directory of Open Access Journals (Sweden)

    Jakub Drozak

    Full Text Available Anserine (beta-alanyl-N(Pi-methyl-L-histidine, a naturally occurring derivative of carnosine (beta-alanyl-L-histidine, is an abundant constituent of skeletal muscles and brain of many vertebrates. Although it has long been proposed to serve as a proton buffer, radicals scavenger and transglycating agent, its physiological function remains obscure. The formation of anserine is catalyzed by carnosine N-methyltransferase which exhibits unknown molecular identity. In the present investigation, we have purified carnosine N-methyltransferase from chicken pectoral muscle about 640-fold until three major polypeptides of about 23, 26 and 37 kDa coeluting with the enzyme were identified in the preparation. Mass spectrometry analysis of these polypeptides resulted in an identification of histamine N-methyltransferase-like (HNMT-like protein as the only meaningful candidate. Analysis of GenBank database records indicated that the hnmt-like gene might be a paralogue of histamine N-methyltransferase gene, while comparison of their protein sequences suggested that HNMT-like protein might have acquired a new activity. Chicken HNMT-like protein was expressed in COS-7 cells, purified to homogeneity, and shown to catalyze the formation of anserine as confirmed by both chromatographic and mass spectrometry analysis. Both specificity and kinetic studies carried out on the native and recombinant enzyme were in agreement with published data. Particularly, several compounds structurally related to carnosine, including histamine and L-histidine, were tested as potential substrates for the enzyme, and carnosine was the only methyl group acceptor. The identification of the gene encoding carnosine N-methyltransferase might be beneficial for estimation of the biological functions of anserine.

  11. Effect of vetch wheat mixture and broccoli as preceding crops on organic summer vegetables: on farm trial in western Turkey

    OpenAIRE

    ASLAN, Burcay

    2011-01-01

    Organic farming requires the use of practices such as crop rotation, green manure and compost application instead of chemical compounds to enhance farm productivity. In this study, effects of two pre-crops (vetch-wheat mixture and broccoli) and additional fertilization (AF) strategy (compost and commercial fertilizer) were tested on organic production of two main crops (tomato and zucchini) and on soil fertility. The main aim of this on-farm trial is to evaluate the applicability of the resul...

  12. Fall and spring broccoli yields and weed control under no-till and conventional tillage with overseeded legume living mulches

    OpenAIRE

    Infante, Michelle Louise

    1994-01-01

    Experiments were conducted with 'BigSur' broccoli (Brassica oleracea var. italica Plenck) at two sites in fall of 1993 and at two sites in the spring of 1994 on a Hayter loam in southwestern Virginia. Objectives were to determine the tillage main-plot (CT = conventional tillage, and NT = no-tillage) and weed-control subplot (C = no overseeding or preemergent herbicide; HB = oxyfluorfen; RC = red clover (Trifolium pratense L.); WC = Dutch white clover (Trifolium repens L.); and ...

  13. The suppressive effect of a commercial extract from Durvillaea potatorum and Ascophyllum nodosum on infection of broccoli by Plasmodiophora brassicae

    OpenAIRE

    Wite, D.; Mattner, S. W.; Porter, I. J.; Arioli, T.

    2015-01-01

    A sand solution technique demonstrated the capacity for a commercial seaweed extract from Durvillaea potatorum and Ascophyllum nodosum (Seasol Commercial®) to significantly suppress infection of broccoli by Plasmodiophora brassicae. In the primary stages of infection, the extract reduced the number of plasmodia formed in the root hairs by 55 %. Later, in the secondary stages of infection, the extract reduced plasmodia in the root cortical cells by up to 84 %. The suppression of infection was ...

  14. Methyl jasmonate and 1-methylcyclopropene treatment effects on quinone reductase inducing activity and post-harvest quality of broccoli.

    Directory of Open Access Journals (Sweden)

    Kang Mo Ku

    Full Text Available Effect of pre-harvest methyl jasmonate (MeJA and post-harvest 1-methylcyclopropene (1-MCP treatments on broccoli floret glucosinolate (GS concentrations and quinone reductase (QR, an in vitro anti-cancer biomarker inducing activity were evaluated two days prior to harvest, at harvest and at 10, 20, and 30 days of post-harvest storage at 4 °C. MeJA treatments four days prior to harvest of broccoli heads was observed to significantly increase floret ethylene biosynthesis resulting in chlorophyll catabolism during post-harvest storage and reduced product quality. Post-harvest treatment with 1-methylcyclopropene (1-MCP, which competitively binds to protein ethylene receptors, maintained post-harvest floret chlorophyll concentrations and product visual quality in both control and MeJA-treated broccoli. Transcript abundance of BoPPH, a gene which is responsible for the synthesis of pheophytinase, the primary enzyme associated with chlorophyll catabolism in broccoli, was reduced by 1-MCP treatment and showed a significant, negative correlation with floret chlorophyll concentrations. The GS, glucobrassicin, neoglucobrassicin, and gluconasturtiin were significantly increased by MeJA treatments. The products of some of the GS from endogenous myrosinase hydrolysis [sulforaphane (SF, neoascorbigen (NeoASG, N-methoxyindole-3-carbinol (NI3C, and phenethyl isothiocyanate (PEITC] were also quantified and found to be significantly correlated with QR. Sulforaphane, the isothiocyanate hydrolysis product of the GS glucoraphanin, was found to be the most potent QR induction agent. Increased sulforaphane formation from the hydrolysis of glucoraphanin was associated with up-regulated gene expression of myrosinase (BoMyo and the myrosinase enzyme co-factor gene, epithiospecifier modifier1 (BoESM1. This study demonstrates the combined treatment of MeJA and 1-MCP increased QR activity without post-harvest quality loss.

  15. Continued Selenium Biofortification of Carrots and Broccoli Grown in Soils Once Amended with Se-enriched S. pinnata.

    Science.gov (United States)

    Bañuelos, Gary S; Arroyo, Irvin S; Dangi, Sadikshya R; Zambrano, Maria C

    2016-01-01

    Selenium (Se) biofortification has been practiced in Se-deficient regions throughout the world primarily by adding inorganic sources of Se to the soil. Considering the use of adding organic sources of Se could be useful as an alternative Se amendment for the production of Se-biofortified food crops. In this multi-year micro-plot study, we investigate growing carrots and broccoli in soils that had been previously amended with Se-enriched Stanleya pinnata Pursh (Britton) three and 4 years prior to planting one and two, respectively. Results showed that total and extractable Se concentrations in soils (0-30 cm) were 1.65 mg kg(-1) and 88 μg L(-1), and 0.92 mg kg(-1) and 48.6 μg L(-1) at the beginning of the growing season for planting one and two, respectively. After each respective growing season, total Se concentrations in the broccoli florets and carrots ranged from 6.99 to 7.83 mg kg(-1) and 3.15 to 6.25 mg kg(-1) in planting one and two, respectively. In broccoli and carrot plant tissues, SeMet (selenomethionine) was the predominant selenoamino acid identified in Se aqueous extracts. In postharvest soils from planting one, phospholipid fatty acid (PLFA) analyses showed that amending the soil with S. pinnata exerted no effect on the microbial biomass, AMF (arbuscular mycorrhizal fungi), actinomycetes and Gram-positive and bacterial PLFA at both 0-5 and 0-30 cm, respectively, 3 years later. Successfully producing Se-enriched broccoli and carrots 3 and 4 years later after amending soil with Se-enriched S. pinnata clearly demonstrates its potential source as an organic Se enriched fertilizer for Se-deficient regions. PMID:27602038

  16. Broccoli and watercress suppress matrix metalloproteinase-9 activity and invasiveness of human MDA-MB-231 breast cancer cells

    International Nuclear Information System (INIS)

    A high dietary intake of cruciferous vegetables has been associated with a reduction in numerous human pathologies particularly cancer. In the current study, we examined the inhibitory effects of broccoli (Brassica oleracea var. italica) and watercress (Rorripa nasturtium aquaticum) extracts on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced cancer cell invasion and matrix metalloproteinase-9 activity using human MDA-MB-231 breast cancer cells. Aberrant overexpression of matrix metalloproteinases, including metalloproteinase-9, is associated with increased invasive potential in cancer cell lines. Our results demonstrate that extracts of broccoli and Rorripa suppressed TPA-induced MMP-9 activity and invasiveness in a concentration dependant manner as determined by zymographic analysis. Furthermore, fractionation of individual extracts followed by liquid chromatography mass spectroscopy analysis (LC-MS) revealed that the inhibitory effects of each vegetable were associated with the presence of 4-methysulfinylbutyl (sulforaphane) and 7-methylsulphinylheptyl isothiocyanates. Taken together, our data indicate that isothiocyanates derived form broccoli and Rorripa inhibit metalloproteinase 9 activities and also suppress the invasive potential of human MDA-MB-231 breast cancer cells in vitro. The inhibitory effects observed in the current study may contribute to the suppression of carcinogenesis by diets high in cruciferous vegetables

  17. Effects of broccoli extract on biodistribution and labeling blood components with {sup 99m}Tc-GH

    Energy Technology Data Exchange (ETDEWEB)

    Cekic, Betul; Muftuler, Fazilet Zumrut Biber; Kilcar, Ayfer Yurt; Ichedef, Cigdem; Unak, Perihan [Ege University, Izmir (Turkey). Inst. of Nuclear Sciences. Dept. of Nuclear Applications

    2011-09-15

    Purpose: people consume vegetables without the knowledge of the side effects of the biological and chemical contents and interactions between radiopharmaceuticals and herbal extract. To this end, current study is focused on the effects of broccoli extract on biodistribution of radiolabeled glucoheptonate ({sup 99m}Tc-GH) and radiolabeling of blood components. Methods: GH was labeled with {sup 99m}Tc. Quality control studies were done utilizing TLC method. Biodistribution studies were performed on male rats which were treated via gavage with either broccoli extract or SF as control group for 15 days. Blood samples were withdrawn from rats' heart. Radiolabeling of blood constituents performed incubating with GH, SnCl{sub 2} and {sup 99m} Tc. Results: radiochemical yield of {sup 99m}Tc-GH is 98.46{+-}1.48 % (n=8). Biodistribution studies have shown that according to the control, the treated group with broccoli has approximately 10 times less uptake in kidney. The percentage of the radioactivity ratios of the blood components is found to be same in both groups. Conclusions: although there is no considerable effect on the radiolabeling of blood components, there is an outstanding change on the biodistribution studies especially on kidneys. The knowledge of this change on kidney uptake may contribute to reduce the risk of misdiagnosis and/or repetition of the examinations in Nuclear Medicine. (author)

  18. EFFECT OF SHADING ON INCIDENCE OF BLACK ROT DISEASE AND YIELD QUALITY OF BROCCOLI IN HUMID TROPICAL REGIONS OF THAILAND

    Directory of Open Access Journals (Sweden)

    Karistsapol Nooprom

    2013-08-01

    Full Text Available The study was carried out in the experimental location of Department of Plant Science, Faculty of Natural Resources, Prince of Songkla University, Thailand, from April to June, 2012. Split-plot in randomized complete block design was used in this experiment with four replications. The broccoli plants under the shading had the lowest incidence of black rot disease (14.48% which was significantly lower than the full sunlight (76.91%. There was a positive impact of the shading on yield quality of the broccoli. The Yok Kheo had the highest head length and diameter (39.13 cm and 12.11 cm, respectively when planting under the shading. These results consistent with broccoli total yield, the highest total yield was obtained from the Yok Kheo under the both shading and full sunlight (10.92 t.ha-1 and 8.29 t.ha-1, respectively, followed by the Green Queen under the shading (6.21 t.ha-1. Therefore, the appropriate variety for planting during dry season in humid tropical regions of Thailand was the Yok Kheo and Green Queen because its total yield per hectare was higher than of the Top Green and they also had higher quality of head.

  19. Functional Role of G9a Histone Methyltransferase in Cancer

    OpenAIRE

    Casciello, Francesco; Windloch, Karolina; Gannon, Frank; Lee, Jason S.

    2015-01-01

    Post-translational modifications of DNA and histones are epigenetic mechanisms, which affect the chromatin structure, ultimately leading to gene expression changes. A number of different epigenetic enzymes are actively involved in the addition or the removal of various covalent modifications, which include acetylation, methylation, phosphorylation, ubiquitination, and sumoylation. Deregulation of these processes is a hallmark of cancer. For instance, G9a, a histone methyltransferase responsib...

  20. Epigenetic Regulation of Autophagy by the Methyltransferase G9a

    OpenAIRE

    Artal-Martinez de Narvajas, Amaia; Gomez, Timothy S.; Zhang, Jin-San; Mann, Alexander O.; Taoda, Yoshiyuki; Gorman, Jacquelyn A.; Herreros-Villanueva, Marta; Gress, Thomas M; Ellenrieder, Volker; Bujanda, Luis; Kim, Do-Hyung; Kozikowski, Alan P.; Koenig, Alexander; Billadeau, Daniel D.

    2013-01-01

    Macroautophagy is an evolutionarily conserved cellular process involved in the clearance of proteins and organelles. Although the cytoplasmic machinery that orchestrates autophagy induction during starvation, hypoxia, or receptor stimulation has been widely studied, the key epigenetic events that initiate and maintain the autophagy process remain unknown. Here we show that the methyltransferase G9a coordinates the transcriptional activation of key regulators of autophagosome formation by remo...

  1. Histamine N-methyltransferase Modulates Human Bronchial Smooth Muscle Contraction

    OpenAIRE

    Tamaoki, J.; Chiyotani, A.; Tagaya, E; Isono, K; Konno, K

    1994-01-01

    To elucidate the modulatory role of histamine-degrading enzymes in airway constrictor responses, human bronchial strips were studied under isometric conditions in vitro. Pretreatment of tissues with the histamine N-methyltransferase (HMT) inhibitor SKF 91488 specifically potentiated the contractile responses to histamine, causing a leftward displacement of the concentration response curves, whereas the diamine oxidase inhibitor aminoguanidine had no effect. This potentiation was attenuated by...

  2. Betaine-homocysteine methyltransferase: human liver genotype-phenotype correlation

    OpenAIRE

    Feng, Qiping; Kalari, Krishna; Fridley, Brooke L.; Jenkins, Gregory; Ji, Yuan; Abo, Ryan; Hebbring, Scott; Zhang, Jianping; Nye, Monica D.; Leeder, J Steven; Weinshilboum, Richard M.

    2010-01-01

    Betaine-homocysteine methyltransferase (BHMT) catalyzes the remethylation of homocysteine. BHMT is highly expressed in the human liver. In the liver, BHMT catalyzes up to 50% of homocysteine metabolism. Understanding the relationship between BHMT genetic polymorphisms and function might increase our understanding of the role of this reaction in homocysteine remethylation and in S-adenosylmethionine-dependent methylation. To help achieve those goals, we measured levels of BHMT enzyme activity ...

  3. Plant isoflavone and isoflavanone O-methyltransferase genes

    Science.gov (United States)

    Broeckling, Bettina E.; Liu, Chang-Jun; Dixon, Richard A.

    2014-08-19

    The invention provides enzymes that encode O-methyltransferases (OMTs) from Medicago truncatula that allow modification to plant (iso)flavonoid biosynthetic pathways. In certain aspects of the invention, the genes encoding these enzymes are provided. The invention therefore allows the modification of plants for isoflavonoid content. Transgenic plants comprising such enzymes are also provided, as well as methods for improving disease resistance in plants. Methods for producing food and nutraceuticals, and the resulting compositions, are also provided.

  4. Structural characterization of the mitomycin 7-O-methyltransferase

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Shanteri; Chang, Aram; Goff, Randal D.; Bingman, Craig A.; Grüschow, Sabine; Sherman, David H.; Phillips, Jr., George N.; Thorson, Jon S. (Michigan); (UW)

    2014-10-02

    Mitomycins are quinone-containing antibiotics, widely used as antitumor drugs in chemotherapy. Mitomycin-7-O-methyltransferase (MmcR), a key tailoring enzyme involved in the biosynthesis of mitomycin in Streptomyces lavendulae, catalyzes the 7-O-methylation of both C9{beta}- and C9{alpha}-configured 7-hydroxymitomycins. We have determined the crystal structures of the MmcR-S-adenosylhomocysteine (SAH) binary complex and MmcR-SAH-mitomycin A (MMA) ternary complex at resolutions of 1.9 and 2.3 {angstrom}, respectively. The study revealed MmcR to adopt a common S-adenosyl-L-methionine-dependent O-methyltransferase fold and the presence of a structurally conserved active site general acid-base pair is consistent with a proton-assisted methyltransfer common to most methyltransferases. Given the importance of C7 alkylation to modulate mitomycin redox potential, this study may also present a template toward the future engineering of catalysts to generate uniquely bioactive mitomycins.

  5. DNA repair methyltransferase (Mgmt) knockout mice are sensitive to the lethal effects of chemotherapeutic alkylating agents.

    OpenAIRE

    Glassner, Brian; Weeda, Geert; Allan, James; Broekhof, Jose'; Carls, Nick; Donker, Ingrid; Engelward, Bevin; Hampson, Richard; Hersmus, Remko; Hickman, Mark; Roth, Richard; Warren, Henry; Wu, Mavis; Hoeijmakers, Jan; Samson, Leona

    1999-01-01

    textabstractWe have generated mice deficient in O6-methylguanine DNA methyltransferase activity encoded by the murine Mgmt gene using homologous recombination to delete the region encoding the Mgmt active site cysteine. Tissues from Mgmt null mice displayed very low O6-methylguanine DNA methyltransferase activity, suggesting that Mgmt constitutes the major, if not the only, O6-methylguanine DNA methyltransferase. Primary mouse embryo fibroblasts and bone marrow cells from Mgmt -/- mice were s...

  6. Molecular Mechanism of Action of Plant DRM De Novo DNA Methyltransferases

    OpenAIRE

    Zhong, Xuehua; Du, Jiamu; Hale, Christopher J.; Gallego-Bartolome, Javier; Feng, Suhua; Vashisht, Ajay A.; Chory, Joanne; Wohlschlegel, James A.; Patel, Dinshaw J.; Jacobsen, Steven E.

    2014-01-01

    DNA methylation is a conserved epigenetic gene regulation mechanism. DOMAINS REARRANGED METHYLTRANSFERASE (DRM) is a key de novo methyltransferase in plants, but how DRM acts mechanistically is poorly understood. Here, we report the crystal structure of the methyltransferase domain of tobacco DRM (NtDRM) and reveal a molecular basis for its rearranged structure. NtDRM forms a functional homo-dimer critical for catalytic activity. We also show that Arabidopsis DRM2 exists in complex with the s...

  7. Comparation of Salt Stress Resistance between Broccoli and Cauliflower%盐胁迫下西兰花和花椰菜抗逆性比较

    Institute of Scientific and Technical Information of China (English)

    蒋海燕

    2015-01-01

    Comparation of salt stress resistance between Broccoli and cauliflower was studied in this paper. The results showed that the concentration of NaCl that significantly inhibited seedling growth of broccoli and cauliflower was 300 mmol/L and 100 mmol/L, respectively. Under the salt stress, the proline content, SOD activity and POD activity of broccoli were higher than that of cauliflower, while the MDA content of broccoli were lower than that of cauliflower, which showed the physiological basis of higher salt resistance in broccoli.%研究比较盐胁迫下西兰花和花椰菜的抗盐性大小。结果表明,NaCl胁迫显著(P<0.05)抑制西兰花幼苗生长的浓度为300 mmol/L;显著抑制花椰菜幼苗生长的浓度为100 mmol/L。盐胁迫下,西兰花脯氨酸含量和SOD、POD活性均较花椰菜高,而丙二醛含量较花椰菜低。结果表明盐胁迫下保护酶活性增强和脯氨酸积累是西兰花具有较强抗盐能力的重要生理学基础。

  8. Structural Biology of Human H3K9 Methyltransferases

    Energy Technology Data Exchange (ETDEWEB)

    Wu, H.; Min, J; Lunin, V; Antoshenko, T; Dombrovsk, L; Zeng, H; Allali-Hassani, A; Campagna-Slater, V; Vedadi, M; et. al.

    2010-01-01

    SET domain methyltransferases deposit methyl marks on specific histone tail lysine residues and play a major role in epigenetic regulation of gene transcription. We solved the structures of the catalytic domains of GLP, G9a, Suv39H2 and PRDM2, four of the eight known human H3K9 methyltransferases in their apo conformation or in complex with the methyl donating cofactor, and peptide substrates. We analyzed the structural determinants for methylation state specificity, and designed a G9a mutant able to tri-methylate H3K9. We show that the I-SET domain acts as a rigid docking platform, while induced-fit of the Post-SET domain is necessary to achieve a catalytically competent conformation. We also propose a model where long-range electrostatics bring enzyme and histone substrate together, while the presence of an arginine upstream of the target lysine is critical for binding and specificity. Post-translational modifications of histone proteins regulate chromatin compaction, mediate epigenetic regulation of transcription, and control cellular differentiation in health and disease. Methylation of histone tails is one of the fundamental events of epigenetic signaling. Tri-methylation of lysine 9 of histone 3 (H3K9) mediates chromatin recruitment of HP1, heterochromatin condensation and gene silencing. Similarly, methylation of H3K27 and H4K20 are associated with a repressed state of chromatin, whereas expressed genes are methylated at H3K4, H3K36 and H3K79. Histone methyltransferases are divided into protein arginine methyltransferases (PRMTs) and histone lysine methyltransferases (HKMTs). HKMTs catalyze the transfer of a methyl group from the co-factor S-adenosyl-L-methionine (SAM) to a substrate lysine and, with the exception of DOT1L, are all organized around a canonical SET domain. The structures of a number of HKMTs have been reported, including ternary complexes of human orthologs with co-factor and substrate peptides (SETD7-H3K4, SETD8-H4K20 and MLL1-H3K4), as well

  9. A SABATH Methyltransferase from the moss Physcomitrella patens catalyzes

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Nan [ORNL; Ferrer, Jean-Luc [Universite Joseph Fourier, France; Moon, Hong S [Department of Plant Sciences, University of Tennessee; Kapteyn, Jeremy [Institute of Biological Chemistry, Washington State University; Zhuang, Xiaofeng [Department of Plant Sciences, University of Tennessee; Hasebe, Mitsuyasu [Laboratory of Evolutionary Biology, National Institute for Biology, 38 Nishigounaka; Stewart, Neal C. [Department of Plant Sciences, University of Tennessee; Gang, David R. [Institute of Biological Chemistry, Washington State University; Chen, Feng [University of Tennessee, Knoxville (UTK)

    2012-01-01

    Known SABATH methyltransferases, all of which were identified from seed plants, catalyze methylation of either the carboxyl group of a variety of low molecular weight metabolites or the nitrogen moiety of precursors of caffeine. In this study, the SABATH family from the bryophyte Physcomitrella patens was identified and characterized. Four SABATH-like sequences (PpSABATH1, PpSABATH2, PpSABATH3, and PpSABATH4) were identified from the P. patens genome. Only PpSABATH1 and PpSABATH2 showed expression in the leafy gametophyte of P. patens. Full-length cDNAs of PpSABATH1 and PpSABATH2 were cloned and expressed in soluble form in Escherichia coli. Recombinant PpSABATH1 and PpSABATH2 were tested for methyltransferase activity with a total of 75 compounds. While showing no activity with carboxylic acids or nitrogen-containing compounds, PpSABATH1 displayed methyltransferase activity with a number of thiols. PpSABATH2 did not show activity with any of the compounds tested. Among the thiols analyzed, PpSABATH1 showed the highest level of activity with thiobenzoic acid with an apparent Km value of 95.5 lM, which is comparable to those of known SABATHs. Using thiobenzoic acid as substrate, GC MS analysis indicated that the methylation catalyzed by PpSABATH1 is on the sulfur atom. The mechanism for S-methylation of thiols catalyzed by PpSABATH1 was partially revealed by homology-based structural modeling. The expression of PpSABATH1 was induced by the treatment of thiobenzoic acid. Further transgenic studies showed that tobacco plants overexpressing PpSABATH1 exhibited enhanced tolerance to thiobenzoic acid, suggesting that PpSABATH1 have a role in the detoxification of xenobiotic thiols.

  10. Archease from Pyrococcus abyssi improves substrate specificity and solubility of a tRNA m5C methyltransferase

    DEFF Research Database (Denmark)

    Auxilien, Sylvie; El Khadali, Fatima; Rasmussen, Anette;

    2007-01-01

    reading frame (PAB1947), which is shown here to encode a tRNA m(5)C methyltransferase. In vitro, the purified recombinant methyltransferase catalyzes m(5)C formation at several cytosines within tRNAs with preference for C49. The specificity of the methyltransferase is increased by the archease...

  11. Effects of combined treatments of irradiation and antimicrobial coatings on reduction of food pathogens in broccoli florets

    International Nuclear Information System (INIS)

    The effect of combined treatment of antimicrobial coatings and γ-radiation on reduction of food pathogens such as Listeria monocytogenes, Escherichia coli, and Salmonella Typhimurium was evaluated in broccoli florets. Broccoli florets were inoculated with pathogenic bacteria at 106 CFU/g. Inoculated florets were then coated with methylcellulose-based coating containing various mixtures of antimicrobial agents: organic acids (OAs) plus lactic acid bacteria metabolites (LABs), OA plus citrus extract (CE), OA plus CE plus spice mixture (SM), and OA plus rosemary extract (RE). Coated florets were irradiated with various doses (0-3.3 kGy), and microbial analyses were used to calculate the D10 value and radiosensitive relative. The coating containing OA plus CE was the most effective formulation for increasing the sensitization of Escherichia coli by 2.4 times as compared to the control without the antimicrobial coating. For Salmonella Typhimurium, coating containing OA plus LAB was the most effective formulation, increasing radiosensitivity by 2.4 times as well. All antimicrobial coatings had almost the same effect of increasing the sensitivity of Listeria monocytogenes (from 1.31 to 1.45 times) to γ-irradiation. - Highlights: → Demonstrate scientifically the synergistic effect of the combined treatment of gamma-irradiation and natural antimicrobial coating in reduction of food pathogens in broccoli. → The coating containing organic acids plus citrus extract was the most efficient formulation for increasing the sensitization of E. coli by 2.40 times as compared to the control. → The coating containing organic acids plus lactic acid bacteria metabolites was the most effective formulation causing the sensitization of S. Typhimurium to γ-irradiation by 2.4 times. → Potential in application of developed formulations to protect food products against food pathogens.

  12. Effects of combined treatments of irradiation and antimicrobial coatings on reduction of food pathogens in broccoli florets

    Energy Technology Data Exchange (ETDEWEB)

    Takala, P.N.; Salmieri, S.; Vu, K.D. [INRS-Institut Armand-Frappier, Research Laboratories in Sciences Applied to Food, Canadian Irradiation Centre, 531, Blvd des Prairies, Laval, QC, H7V 1B7 (Canada); Lacroix, M., E-mail: Monique.Lacroix@iaf.inrs.ca [INRS-Institut Armand-Frappier, Research Laboratories in Sciences Applied to Food, Canadian Irradiation Centre, 531, Blvd des Prairies, Laval, QC, H7V 1B7 (Canada)

    2011-12-15

    The effect of combined treatment of antimicrobial coatings and {gamma}-radiation on reduction of food pathogens such as Listeria monocytogenes, Escherichia coli, and Salmonella Typhimurium was evaluated in broccoli florets. Broccoli florets were inoculated with pathogenic bacteria at 10{sup 6} CFU/g. Inoculated florets were then coated with methylcellulose-based coating containing various mixtures of antimicrobial agents: organic acids (OAs) plus lactic acid bacteria metabolites (LABs), OA plus citrus extract (CE), OA plus CE plus spice mixture (SM), and OA plus rosemary extract (RE). Coated florets were irradiated with various doses (0-3.3 kGy), and microbial analyses were used to calculate the D{sub 10} value and radiosensitive relative. The coating containing OA plus CE was the most effective formulation for increasing the sensitization of Escherichia coli by 2.4 times as compared to the control without the antimicrobial coating. For Salmonella Typhimurium, coating containing OA plus LAB was the most effective formulation, increasing radiosensitivity by 2.4 times as well. All antimicrobial coatings had almost the same effect of increasing the sensitivity of Listeria monocytogenes (from 1.31 to 1.45 times) to {gamma}-irradiation. - Highlights: > Demonstrate scientifically the synergistic effect of the combined treatment of gamma-irradiation and natural antimicrobial coating in reduction of food pathogens in broccoli. > The coating containing organic acids plus citrus extract was the most efficient formulation for increasing the sensitization of E. coli by 2.40 times as compared to the control. > The coating containing organic acids plus lactic acid bacteria metabolites was the most effective formulation causing the sensitization of S. Typhimurium to {gamma}-irradiation by 2.4 times. > Potential in application of developed formulations to protect food products against food pathogens.

  13. Organelle Simple Sequence Repeat Markers Help to Distinguish Carpelloid Stamen and Normal Cytoplasmic Male Sterile Sources in Broccoli.

    Science.gov (United States)

    Shu, Jinshuai; Liu, Yumei; Li, Zhansheng; Zhang, Lili; Fang, Zhiyuan; Yang, Limei; Zhuang, Mu; Zhang, Yangyong; Lv, Honghao

    2015-01-01

    We previously discovered carpelloid stamens when breeding cytoplasmic male sterile lines in broccoli (Brassica oleracea var. italica). In this study, hybrids and multiple backcrosses were produced from different cytoplasmic male sterile carpelloid stamen sources and maintainer lines. Carpelloid stamens caused dysplasia of the flower structure and led to hooked or coiled siliques with poor seed setting, which were inherited in a maternal fashion. Using four distinct carpelloid stamens and twelve distinct normal stamens from cytoplasmic male sterile sources and one maintainer, we used 21 mitochondrial simple sequence repeat (mtSSR) primers and 32 chloroplast SSR primers to identify a mitochondrial marker, mtSSR2, that can differentiate between the cytoplasm of carpelloid and normal stamens. Thereafter, mtSSR2 was used to identify another 34 broccoli accessions, with an accuracy rate of 100%. Analysis of the polymorphic sequences revealed that the mtSSR2 open reading frame of carpelloid stamen sterile sources had a deletion of 51 bases (encoding 18 amino acids) compared with normal stamen materials. The open reading frame is located in the coding region of orf125 and orf108 of the mitochondrial genomes in Brassica crops and had the highest similarity with Raphanus sativus and Brassica carinata. The current study has not only identified a useful molecular marker to detect the cytoplasm of carpelloid stamens during broccoli breeding, but it also provides evidence that the mitochondrial genome is maternally inherited and provides a basis for studying the effect of the cytoplasm on flower organ development in plants. PMID:26407159

  14. Human placental DNA methyltransferase: DNA substrate and DNA binding specificity.

    OpenAIRE

    Wang, R.Y.; Huang, L. H.; Ehrlich, M

    1984-01-01

    We have partially purified a DNA methyltransferase from human placenta using a novel substrate for a highly sensitive assay of methylation of hemimethylated DNA. This substrate was prepared by extensive nick translation of bacteriophage XP12 DNA, which normally has virtually all of its cytosine residues replaced by 5-methylcytosine (m5C). Micrococcus luteus DNA was just as good a substrate if it was first similarly nick translated with m5dCTP instead of dCTP in the polymerization mixture. At ...

  15. Expanding Cofactor Repertoire of Protein Lysine Methyltransferase for Substrate Labeling

    OpenAIRE

    Islam, Kabirul; Zheng, Weihong; Yu, Haiqiang; Deng, Haiteng; Luo, Minkui

    2011-01-01

    Protein lysine methyltransferases (PKMTs) play crucial roles in normal physiology and disease processes. Profiling PKMT targets is an important but challenging task. With cancer-relevant G9a as a target, we have demonstrated the success in developing S-adenosyl-L-methionine (SAM) analogues, particularly (E)-hex-2-en-5-ynyl SAM (Hey-SAM), as cofactors for engineered G9a. Hey-SAM analogue in combination with G9a Y1154A mutant modifies the same set of substrates as their native counterparts with...

  16. Purification and characterization of DNA methyltransferases from Neisseria gonorrhoeae.

    OpenAIRE

    Piekarowicz, A; Yuan, R.; Stein, D C

    1988-01-01

    Three DNA methyltransferases, M.NgoAI, and M.NgoBI and M.NgoBII, free of any nuclease activities were isolated from Neisseria gonorrhoeae strains WR220 and MUG116 respectively. M.NgoAI recognizes the sequence 5' GGCC 3' and methylates the first 5' cytosine on both strands. M.NgoBI and M.NgoBII recognize 5' TCACC 3' and 5' GTAN5CTC 3' respectively. M.NgoBII methylates cytosine on only one strand to produce 5' GTAN5mCTC 3'.

  17. High level expression and purification of HhaI methyltransferase.

    OpenAIRE

    Wu, J. C.; Santi, D V

    1988-01-01

    A cloning system for the DNA-(cytosine-5)-methyltransferase MHhaI and high level expression of the enzyme are described. A parent plasmid was constructed from fragments of the MHhaI gene and synthetic oligonucleotides. The construct permits introduction of various restriction sites for cloning at precise positions near the initiation codon, and beyond the termination codon. The entire MHhaI coding sequence was introduced as a 1042 b.p. NdeI-XbaI fragment into the vector pAR3040 which contains...

  18. Catalytic site remodelling of the DOT1L methyltransferase by selective inhibitors

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Wenyu [Univ. of Toronto, ON (Canada); Chory, Emma J. [Dana-Farber Cancer Inst., Boston, MA (United States); Northeastern Univ., Boston, MA (United States); Wernimont, Amy K. [Univ. of Toronto, ON (Canada); Tempel, Wolfram [Univ. of Toronto, ON (Canada); Scopton, Alex [Univ. of Toronto, ON (Canada); Federation, Alexander [Dana-Farber Cancer Inst., Boston, MA (United States); Marineau, Jason J. [Dana-Farber Cancer Inst., Boston, MA (United States); Qi, Jun [Dana-Farber Cancer Inst., Boston, MA (United States); Barsyte-Lovejoy, Dalia [Univ. of Toronto, ON (Canada); Yi, Joanna [Dana-Farber Cancer Inst., Boston, MA (United States); Marcellus, Richard [Ontario Inst. for Cancer Research, Toronto, ON (Canada); Iacob, Roxana E. [Northeastern Univ., Boston, MA (United States); Engen, John R. [Northeastern Univ., Boston, MA (United States); Griffin, Carly [Ontario Inst. for Cancer Research, Toronto, ON (Canada); Aman, Ahmed [Ontario Inst. for Cancer Research, Toronto, ON (Canada); Wienholds, Erno [Univ. of Toronto, ON (Canada); Li, Fengling [Univ. of Toronto, ON (Canada); Pineda, Javier [Dana-Farber Cancer Inst., Boston, MA (United States); Univ. of Notre Dame, IN (United States); Estiu, Guillermina [Univ. of Notre Dame, IN (United States); Shatseva, Tatiana [Univ. of Toronto, ON (Canada); Hajian, Taraneh [Univ. of Toronto, ON (Canada); Al-awar, Rima [Ontario Inst. for Cancer Research, Toronto, ON (Canada); Dick, John E. [Univ. of Toronto, ON (Canada); Vedadi, Masoud [Univ. of Toronto, ON (Canada); Brown, Peter J. [Univ. of Toronto, ON (Canada); Arrowsmith, Cheryl H. [Univ. of Toronto, ON (Canada); Bradner, James E. [Dana-Farber Cancer Inst., Boston, MA (United States); Harvard Medical School, Boston, MA (United States); Schapira, Matthieu [Univ. of Toronto, ON (Canada)

    2012-12-18

    Selective inhibition of protein methyltransferases is a promising new approach to drug discovery. An attractive strategy towards this goal is the development of compounds that selectively inhibit binding of the cofactor, S-adenosylmethionine, within specific protein methyltransferases. Here we report the three-dimensional structure of the protein methyltransferase DOT1L bound toEPZ004777, the first S-adenosylmethionine-competitive inhibitor of a protein methyltransferase with in vivo efficacy. This structure and those of four new analogues reveal remodelling of the catalytic site. EPZ004777 and a brominated analogue, SGC0946, inhibit DOT1L in vitro and selectively kill mixed lineage leukaemia cells, in which DOT1L is aberrantly localized via interaction with an oncogenic MLL fusion protein. These data provide important new insight into mechanisms of cell-active S-adenosylmethionine-competitive protein methyltransferase inhibitors, and establish a foundation for the further development of drug-like inhibitors of DOT1L for cancer therapy.

  19. Dietary Chemoprevention of PhIP Induced Carcinogenesis in Male Fischer 344 Rats with Tomato and Broccoli

    OpenAIRE

    Kirstie Canene-Adams; Sfanos, Karen S; Chung-Tiang Liang; Srinivasan Yegnasubramanian; Nelson, William G.; Cory Brayton; Marzo, Angelo M. De

    2013-01-01

    The heterocyclic amine, 2-amino-1-methyl-6-phenylimidazo[4,5-B]pyridine (PhIP), found in meats cooked at high temperatures, has been implicated in epidemiological and rodent studies for causing breast, prostate, and colorectal cancers. A previous animal study using a xenograft model has shown that whole tomato and broccoli, when eaten in combination, exhibit a marked effect on tumor reduction compared to when eaten alone. Our aim was to determine if PhIP-induced carcinogenesis can be prevente...

  20. Improved synthesis methods of standards used for quantitative determination of total isothiocyanates from broccoli in human urine

    DEFF Research Database (Denmark)

    Kristensen, Mette; Krogholm, Kirstine Suszkiewicz; Frederiksen, Hanne;

    2007-01-01

    A well-known method for quantification of isothiocyanates (ITCs) and their metabolites is the condensation reaction with 1,2-benzenedithiole to produce 1,3-benzodithiole-2-thione, which can be quantified by high-performance liquid chromatography. Standards of an ITC metabolite and 1,3-benzodithio...... excretion of ITCs from 10 healthy subjects who consumed 350 g broccoli. The excretion was investigated throughout 48 h showing a cumulative urinary ITC excretion of 49.1 +/- 25.2% of the dose....

  1. Growth of lowland broccoli coconut coir dust and peat based growing media using fertigation system

    International Nuclear Information System (INIS)

    A study was carried out inside the greenhouse to determine the response of lowland broccoli (Brassica oleracea) plants grown in five mixtures of coconut coir dust and peat as the growing media. The growing mixes were prepared in the following ratios (% by vol): 100% coconut coir dust, 75% coconut coir dust + 25% peat, 50% coconut coir dust + 50% peat, 25% coconut coir dust + 75% peat and 100% peat. The plants were supplied with 200 mg 1-1 nitrogen using a computerized fertigation system developed at MINT Plants grown on 100% peat exhibited reduced plant growth and yield compared to plants grown on mixes containing coconut coir dust. Plants growth and yield were increased on growing media contained > 50% coconut coir dust; however, the highest total plant dry weight, plant height and yield were obtained from plants grown on 100% coconut coir dust. Total nitrogen concentration in the leaves and stems was not significantly different in all mixture of the growing media regardless of whether coconut coir dust or peat was used. In general, nitrate -nitrogen concentrations were lowest in the floret than in the leaves and stem tissues of plants grown on all growing media used in the study. However, nitrate-nitrogen concentrations in the leaves, stems and florets were lowest in the plants grown on 100% peat compared to the plants grown on growing media containing coconut coir dust. The concentrations of nitrate-nitrogen in the florets, which ranged from 290 to 450 mg N kg-l/dry weight, are considered to be under permissible levels by European standards. (Author)

  2. Lipid substrate specificity of phosphatidylethanolamine N-methyltransferase of Tetrahymena

    International Nuclear Information System (INIS)

    The ciliate protozoan Tetrahymena thermophila forms about 60% of its phosphatidylcholine by methylation of phosphatidylethanolamine with S-adenosylmethionine using the enzyme phosphatidylethanolamine N-methyltransferase. Analogues of ethanolamine or of ethanolamine phosphate are incorporated into the phospholipids of Tetrahymena when cells are cultured in their presence. These compounds, 3-amino-1-propanol, 2-aminoethylphosphonate, 3-aminopropylphosphonate and N,N-dimethylaminoethylphosphonate replace from 50 to 75% of the ethanolamine phosphate in phosphatidylethanolamine. However, analysis of the phospholipids of lipid-altered Tetrahymena showed that none of the phosphatidylethanolamine analogues had been converted to the corresponding phosphatidylcholine analogue. No incorration of [14C-CH3]methionine into the phosphatidylcholine analogues could be demonstrated in vivo, nor was label from [3H-CH3]S-adenosylmethionine incorporated in virto. Thus, only phosphatidylethanolamine and its monomethyl and dimethyl derivatives have been found to be substrates for the phosphatidylethanoiamine N-methyltransferase. The enzyme therefore requires a phospholipid substrate containing an ester linkage between the alkylamine and phosphorus, with the amino group required to be β to the alcohol

  3. Hypnotizability and Catechol-O-Methyltransferase (COMT polymorphysms in Italians

    Directory of Open Access Journals (Sweden)

    Alessandro Gialluisi

    2014-01-01

    Full Text Available Higher brain dopamine content depending on lower activity of Catechol-O-Methyltransferase (COMT in subjects with high hypnotisability scores (highs has been considered responsible for their attentional characteristics. However, the results of the previous genetic studies on association between hypnotisability and the Catechol-O-Methyltransferase (COMT single nucleotide polymorphism (SNP rs4680 (Val158Met were inconsistent. Here, we used a selective genotyping approach to re-evaluate the association between hypnotisability and COMT in the context of a two-SNP haplotype analysis, considering not only the Val158Met polymorphism, but also the closely located rs4818 SNP. An Italian sample of 53 highs, 49 low hypnotizable subjects (lows and 57 controls, were genotyped for a segment of 805 bp of the COMT gene, including Val158Met and the closely located rs4818 SNP. Our selective genotyping approach had 97.1% power to detect the previously reported strongest association at the significance level of 5%. We found no evidence of association at the SNP, haplotype and diplotype levels. Thus, our results challenge the dopamine-based theory of hypnosis and indirectly support recent neuropsychological and neurophysiological findings reporting the lack of any association between hypnotisability and focused attention abilities.

  4. Structural biology of human H3K9 methyltransferases.

    Directory of Open Access Journals (Sweden)

    Hong Wu

    Full Text Available UNLABELLED: SET domain methyltransferases deposit methyl marks on specific histone tail lysine residues and play a major role in epigenetic regulation of gene transcription. We solved the structures of the catalytic domains of GLP, G9a, Suv39H2 and PRDM2, four of the eight known human H3K9 methyltransferases in their apo conformation or in complex with the methyl donating cofactor, and peptide substrates. We analyzed the structural determinants for methylation state specificity, and designed a G9a mutant able to tri-methylate H3K9. We show that the I-SET domain acts as a rigid docking platform, while induced-fit of the Post-SET domain is necessary to achieve a catalytically competent conformation. We also propose a model where long-range electrostatics bring enzyme and histone substrate together, while the presence of an arginine upstream of the target lysine is critical for binding and specificity. ENHANCED VERSION: This article can also be viewed as an enhanced version in which the text of the article is integrated with interactive 3D representations and animated transitions. Please note that a web plugin is required to access this enhanced functionality. Instructions for the installation and use of the web plugin are available in Text S1.

  5. Effects of combined treatments of irradiation and antimicrobial coatings on reduction of food pathogens in broccoli florets

    Science.gov (United States)

    Takala, P. N.; Salmieri, S.; Vu, K. D.; Lacroix, M.

    2011-12-01

    The effect of combined treatment of antimicrobial coatings and γ-radiation on reduction of food pathogens such as Listeria monocytogenes, Escherichia coli, and Salmonella Typhimurium was evaluated in broccoli florets. Broccoli florets were inoculated with pathogenic bacteria at 10 6 CFU/g. Inoculated florets were then coated with methylcellulose-based coating containing various mixtures of antimicrobial agents: organic acids (OAs) plus lactic acid bacteria metabolites (LABs), OA plus citrus extract (CE), OA plus CE plus spice mixture (SM), and OA plus rosemary extract (RE). Coated florets were irradiated with various doses (0-3.3 kGy), and microbial analyses were used to calculate the D10 value and radiosensitive relative. The coating containing OA plus CE was the most effective formulation for increasing the sensitization of Escherichia coli by 2.4 times as compared to the control without the antimicrobial coating. For Salmonella Typhimurium, coating containing OA plus LAB was the most effective formulation, increasing radiosensitivity by 2.4 times as well. All antimicrobial coatings had almost the same effect of increasing the sensitivity of Listeria monocytogenes (from 1.31 to 1.45 times) to γ-irradiation.

  6. Effect of Planting Time on Incidence of Bacterial Disease and Yield of Broccoli during Dry Season in Southern Thailand

    Directory of Open Access Journals (Sweden)

    Karistsapol Nooprom

    2013-08-01

    Full Text Available The trial was aimed to select suitable planting times when broccoli varieties could resist well to soft rot and black rot diseases during dry season was studies at Prince of Songkla University, Songkhla, Thailand, from January to June, 2011. Split-plot in randomized complete block design was used in this experiment with four replications. The results showed that the lowest incidence of soft rot disease was observed in all broccoli varieties when they were planted in January (4.19-7.18%. After that, it would increase (22.59-45.58% but the incidence of black rot disease was not increased when planting was done in January, February, March and June. The incidence of black rot disease was highest in April (84.24-92.64%. The Yok Kheo was produced the highest total yield when the planting took place in January and March (12.31 and 10.65 t/ha, respectively. The Yok Kheo is an interesting new hybrid variety. Its higher yield was than Top Green which is popular variety grown in southern Thailand. The yield of Green Queen was not significantly different from Top Green. It can be harvested at 11.67 and 9.38 days earlier than Top Green and Yok Kheo, respectively.

  7. Gold nanoparticles synthesized by Brassica oleracea (Broccoli) acting as antimicrobial agents against human pathogenic bacteria and fungi

    Science.gov (United States)

    Piruthiviraj, Prakash; Margret, Anita; Krishnamurthy, Poornima Priyadharsani

    2016-04-01

    Production of antimicrobial agents through the synthesis of gold nanoparticles using green technology has been extensively made consistent by various researchers; yet, this study uses the flower bud's aqueous extracts of Brassica oleracea (Broccoli) as a reducing agent for chloroauric acid (1 mM). After 30 min of incubation, synthesis of gold nanoparticles (AuNps) was observed by a change in extract color from pale yellow to purple color. Synthesis of AuNps was confirmed in UV-visible spectroscopy at the range of approximately 560 nm. The SEM analysis showed the average nanoparticles size of 12-22 nm. The antimicrobial activity of AuNps was analyzed by subjecting it to human pathogenic bacteria (Gram-positive Staphylococcus aureus and Gram-negative Klebsiella pneumonia) and fungi (Aspergillus flavus, Aspergillus niger and Candida albicans) using disc diffusion method. The broccoli-synthesized AuNps showed the efficient antibacterial and antifungal activity of above-mentioned microbes. It was confirmed that AuNps have the best antimicrobial agent compared to the standard antibiotics (Gentamicin and Fluconazole). When the concentrations of AuNps were increased (10, 25, and 50 µg/ml), the sensitivity zone also increased for all the tested microbes. The synthesized AuNps are capable of rendering high antimicrobial efficacy and, hence, have a great potential in the preparation of drugs used against major bacterial and fungal diseases in humans.

  8. Enhanced resistance to Botrytis cinerea and Rhizoctonia solani in transgenic broccoli with a Trichoderma viride endochitinase gene

    Institute of Scientific and Technical Information of China (English)

    YU Ya; REN Shu-xin; GUO Yang-dong; ZHANG Lei; LIAN Wei-ran; XU Feng-feng; LI Shuang-tao; XIANG Juan; ZHANG Guo-zhen; HU Zan-min; ZHAO Bing

    2015-01-01

    A endochitinase gene (Tch ) from the fungus Trichoderma viride was introduced into broccoli (Brassica oleracea var. italica) by Agrobacterium-mediated transformation. Sixty-eight putative transformants were obtained and the presence of the Tch gene was conifrmed by both PCR and Southern blot analysis. RT-PCR analysis showed an accumulation of the transcript encoding the endochitinase protein in the transgenic plants. Using real-time quantitative PCR, the expression proifling of endochitinase gene was analyzed. Primary transformants and selfed progeny were examined for expression of the endo-chitinase using a lfuorometric assay and for their resistance to the pathogenic fungi Botrytis cinerea and Rhizoctonia solani. The endochitinase activities in T0 in vitro plants, T0 mature plants and T1 mature plants were correlated with leaf lesions, and the transgenic line T618 had high endochitinse activities of 102.68, 114.53 and 120.27 nmol L–1 MU min–1 mg–1 protein in the three kinds of plants, respectively. The endochitinase activity showed a positive correlation with the resistance to the pathogens. Most transgenic T0 broccoli had increased resistance to the pathogens of B. cinerea and R. solani in leaf assays and this resistance was conifrmed to be inheritable. These ifndings suggested that expression of the Tch gene from T. viride could enhance resistance to pathogenic fungi in Brassica species.

  9. Differential Responses of Two Broccoli (Brassica oleracea L. var Italica Cultivars to Salinity and Nutritional Quality Improvement

    Directory of Open Access Journals (Sweden)

    Chokri Zaghdoud

    2012-01-01

    Full Text Available The comparative responses of two broccoli cultivars (Brassica oleracea var. Italica, cv. Parthenon and cv. Naxos to a 15 d exposure to different NaCl levels were investigated. Salinity led to increased concentrations of Na+ and Cl− ions in both cultivars, a disruption of the endogenous minerals levels in the shoots and roots—that varied with the cultivar and salt concentration—and decreases in the osmotic potential (Ψπ, root hydraulic conductance (L0, and stomatal conductance (Gs. The reduced biomass of Naxos at moderate NaCl indicates greater sensitivity to salinity, compared with Parthenon. Parthenon accumulated more soluble sugars, for osmotic adjustment, whereas Naxos accumulated proline, which gave the two cultivars differing nutritional characteristics. The total glucosinolates (GSLs content was not affected by salinity in Parthenon while it decreased significantly in Naxos as a consequence of the decrease in the indole GSL. However, Naxos accumulated more aliphatic GSLs under salt stress than Parthenon, which confers on this cultivar a greater nutritional value when cultivated under salinity.These results suggest that, at distinct salinity levels, each broccoli cultivar adopts a specific strategy, indicating the crucial role of the genetic background on the organoleptic and nutritional properties that each cultivar acquires.

  10. SELENIUM (SE) ENRICHED BEEF, WHEAT OR BROCCOLI OR THE SALT SELENATE DIFFER IN THE ABILITY TO INCREASE THIOREDOXIN REDUCTASE (TR) AND GLUTATHIONE PEROXIDASE (GPX) IN RATS

    Science.gov (United States)

    Wheat and beef are the most abundant sources of Se in a western diet; they contain Se primarily in the form of selenomethionine and selenocysteine. Broccoli is a Se-accumulator that contains large amounts of Se-methyl selenocysteine. Bioavailability of Se from these foods was investigated by measur...

  11. VEGETATIVE GROWTH AND PRODUCTIVITY OF A NUMBER OF INTRODUCED BROCCOLI CULTIVARS (Brassica oleracea L. var. italica Plenck. IN BATUR VILLAGE, KINTAMANI DISTRICT, BANGLI REGENCY, BALI

    Directory of Open Access Journals (Sweden)

    Ni Kadek Raleni

    2015-09-01

    Full Text Available Broccoli (Brassica oleracea L. var. italica Plenck. is a vegetable crop belongs to Brassicaceae family. Broccoli has high nutrition, high in fiber and contains isotiacyanate that has anticancer activity. Broccoli market in Indonesia, particularly in modern supermarkets, increases 15-20% per year, however, productivity was still low, therefore effort to increase broccoli production in Indonesia need to be investigated. Field trial was conducted at Batur Village, Kintamani District, Bangli Regency, Bali, to find out cultivars that were adapted in tropical region. Introduced cultivars being trialed were ‘Alborada’, ‘Belstar’, ‘Fiesta’, ‘Sarasota’, ‘Bay Meadows’, ‘Castle Dome’, ‘Liutenant’, ‘Iron Man’, ‘Marathon’, ‘Green Gold’, ‘Imperial’, ‘Green Magic’ and ‘Lucky’ as control. Variable observed were vegetative growth, curding period, and productivity of each cultivar. This study employed Randomized Completely Block Design with 3 replicates (plots and 8 plants each plot. Data were analyzed using ANOVA (Analysis of variance on Costat program, followed by Duncan’s Multiple Range Test (DMRT on 1% level. Results show that each cultivar varied in adaptability in tropical region. ‘Castle Dome’ has the highest productivity, while ‘Fiesta’ was the lowest.

  12. 冰温贮藏对西兰花保鲜的影响%Effect of ice-temperature storage on the freshness retaining of broccolis

    Institute of Scientific and Technical Information of China (English)

    林本芳; 鲁晓翔; 李江阔; 陈绍慧

    2012-01-01

    研究冰温贮藏对西兰花保鲜效果的影响。将西兰花分为两组,一组存于冰温库中(-0.7~-0.4)℃,另一组存于普通冷库中(-1~1)℃。每14d取样测定其理化指标及营养成分。实验结果表明:冰温贮藏对于延缓西兰花可溶性固形物含量、可溶性糖浓度和VC含量的下降具有明显的效果,可显著抑制西兰花贮藏期间乙烯生成率,延迟乙烯高峰的出现,但对呼吸的抑制效果不大;冰温贮藏提高了西兰花POD、SOD和CAT等的活性,降低了PPO的活性,延缓了西兰花的衰老。这说明西兰花适于冰温贮藏,且冰温贮藏的效果优于普通冷藏的效果。%In order to investigate the effect of ice-temperature storage on the results of freshness retaining of broccolis,the broccolis were stored under two different storage temperatures.One was-0.7~-0.4℃,and the other was-1~1℃.The physicochemical indexes and nutrition components were measured for 14 days.The research results showed that the soluble solids content,soluble sugar and vitamine C of the broccolis by ice-temperature storage decreased less than that of ordinary storage.Ice-temperature storage could significantly inhibit the ethylene generation rate of the broccolis,but the inhibition effect on respiration of the broccolis was not obvious.The activities of peroxidase(POD),superoxide dismutase(SOD)and catalase(CAT)were promoted and the activity of polyphenol oxidas(PPO)was reduced,in turn,the ripening of broccolis was delayed.Therefore,the storage effect of broccolis by ice-temperature storage is superior to that by ordinary storage.

  13. Substrate recognition and modification by the nosiheptide resistance methyltransferase.

    Directory of Open Access Journals (Sweden)

    Sitao Yin

    Full Text Available The proliferation of antibiotic resistant pathogens is an increasing threat to the general public. Resistance may be conferred by a number of mechanisms including covalent or mutational modification of the antibiotic binding site, covalent modification of the drug, or the over-expression of efflux pumps. The nosiheptide resistance methyltransferase (NHR confers resistance to the thiazole antibiotic nosiheptide in the nosiheptide producer organism Streptomyces actuosus through 2'O-methylation of 23S rRNA at the nucleotide A1067. Although the crystal structures of NHR and the closely related thiostrepton-resistance methyltransferase (TSR in complex with the cofactor S-Adenosyl-L-methionine (SAM are available, the principles behind NHR substrate recognition and catalysis remain unclear.We have analyzed the binding interactions between NHR and model 58 and 29 nucleotide substrate RNAs by gel electrophoresis mobility shift assays (EMSA and fluorescence anisotropy. We show that the enzyme binds to RNA as a dimer. By constructing a hetero-dimer complex composed of one wild-type subunit and one inactive mutant NHR-R135A subunit, we show that only one functional subunit of the NHR homodimer is required for its enzymatic activity. Mutational analysis suggests that the interactions between neighbouring bases (G1068 and U1066 and A1067 have an important role in methyltransfer activity, such that the substitution of a deoxy sugar spacer (5' to the target nucleotide achieved near wild-type levels of methylation. A series of atomic substitutions at specific positions on the substrate adenine show that local base-base interactions between neighbouring bases are important for methylation.Taken together these data suggest that local base-base interactions play an important role in aligning the substrate 2' hydroxyl group of A1067 for methyl group transfer. Methylation of nucleic acids is playing an increasingly important role in fundamental biological processes

  14. Changes in proximate composition, biochemical and antioxidant attributes broccoli (brassica oleracea l.) in relation to foliar application of selected plant growth regulators

    International Nuclear Information System (INIS)

    Field experiments were conducted to investigate the effect of foliar application of selected plant growth regulators (PGRs) on the proximate composition, biochemical and antioxidants attributes of broccoli (Brassica oleracea L.) leaves. Different concentrations of exogenously applied PGRs such as humic acid (HA), cytokinin (CK), gibberellic acid (GA3) and GA3+CK exhibited variable effect on the tested parameters. The foliar spray with HA (0.6 percentage), GA3+CK (20+20 ppm), GA3 (20 ppm) and CK (40 ppm) increased the contents of moisture, ash, crude fiber and crude protein in broccoli leaves. With few exceptions, the antioxidant capacity of broccoli leaves, in terms of estimation of total phenolics content (TPC), total flavonoids content (TFC), reducing power and DPPH radical scavenging activity, was also improved due to foliar spray of PGRs. The results showed that maximum contents of total chlorophyll (1.98 mg/g FW), protein (7.09 mg/g FW), and proline (0.62 μg/g FW) were exhibited by GA3+CK (10+10 ppm), GA3 (20 ppm) and GA3+CK (20+20 ppm) treated samples, respectively. Meanwhile, GA3+CK (10+10 ppm) was found to be the most effective growth promoter in lowering melondialdehyde (MDA) content (0.89 micro M/g FW) of broccoli leaves. In conclusion, it can be revealed that optimized foliar spray of selected PGRs, and especially of HA and mixture of GA3 and CK, is practically applicable towards improving biochemical and antioxidant attributes of broccoli leaves with potential nutritional benefits. (author)

  15. Chromosome Doubling of Microspore-Derived Plants from Cabbage (Brassica oleracea var. capitata L.) and Broccoli (Brassica oleracea var. italica L.)

    Science.gov (United States)

    Yuan, Suxia; Su, Yanbin; Liu, Yumei; Li, Zhansheng; Fang, Zhiyuan; Yang, Limei; Zhuang, Mu; Zhang, Yangyong; Lv, Honghao; Sun, Peitian

    2015-01-01

    Chromosome doubling of microspore-derived plants is an important factor in the practical application of microspore culture technology because breeding programs require a large number of genetically stable, homozygous doubled haploid plants with a high level of fertility. In the present paper, 29 populations of microspore-derived plantlets from cabbage (Brassica oleracea var. capitata) and broccoli (Brassica oleracea var. italica) were used to study the ploidy level and spontaneous chromosome doubling of these populations, the artificial chromosome doubling induced by colchicine, and the influence of tissue culture duration on the chromosomal ploidy of the microspore-derived regenerants. Spontaneous chromosome doubling occurred randomly and was genotype dependent. In the plant populations derived from microspores, there were haploids, diploids, and even a low frequency of polyploids and mixed-ploidy plantlets. The total spontaneous doubling in the 14 cabbage populations ranged from 0 to 76.9%, compared with 52.2 to 100% in the 15 broccoli populations. To improve the rate of chromosome doubling, an efficient and reliable artificial chromosome doubling protocol (i.e., the immersion of haploid plantlet roots in a colchicine solution) was developed for cabbage and broccoli microspore-derived haploids. The optimal chromosome doubling of the haploids was obtained with a solution of 0.2% colchicine for 9–12 h or 0.4% colchicine for 3–9 h for cabbage and 0.05% colchicine for 6–12 h for broccoli. This protocol produced chromosome doubling in over 50% of the haploid genotypes for most of the populations derived from cabbage and broccoli. Notably, after 1 or more years in tissue culture, the chromosomes of the haploids were doubled, and most of the haploids turned into doubled haploid or mixed-ploidy plants. This is the first report indicating that tissue culture duration can change the chromosomal ploidy of microspore-derived regenerants. PMID:26734028

  16. Plants as biofactories: Postharvest Stress-Induced Accumulation of Phenolic Compounds and Glucosinolates in Broccoli Subjected to Wounding Stress and Exogenous Phytohormones

    Directory of Open Access Journals (Sweden)

    Daniel eVillarreal-García

    2016-02-01

    Full Text Available Broccoli contains high levels of bioactive molecules and is considered a functional food. In this study, postharvest treatments to enhance the concentration of glucosinolates and phenolic compounds were evaluated. Broccoli whole heads were wounded to obtain florets and wounded florets (florets cut into four even pieces and stored for 24 h at 20 ºC with or without exogenous ethylene (ET, 1000 ppm or methyl jasmonate (MeJA, 250 ppm. Whole heads were used as a control for wounding treatments. Regarding glucosinolate accumulation, ET selectively induced the 4-hydroxylation of glucobrassicin in whole heads, resulting in ~223% higher 4-hydroxyglucobrassicin than time 0 h samples. Additionally, glucoraphanin was increased by ~53% in whole heads treated with ET, while neoglucobrassicin was greatly accumulated in wounded florets treated with ET or MeJA, showing increases of ~193% and ~286%, respectively. On the other hand, although only whole heads stored without phytohormones showed higher concentrations of phenolic compounds, which was reflected in ~33%, ~30%, and 46% higher levels of 1,2,2-trisinapoylgentiobose, 1,2-diferulolylgentiobiose, and 1,2-disinapoyl-2-ferulolylgentiobiose, respectively; broccoli florets stored under air control conditions showed enhanced concentrations of 3-O-caffeoylquinic acid, 1,2-disinapoylgentiobiose, and 1,2-disinapoyl-2-ferulolylgentiobiose (~22%, ~185%, and ~65% more, respectively. However, exogenous ET and MeJA impeded individual phenolics accumulation. Results allowed the elucidation of simple and effective postharvest treatment to enhance the content of individual glucosinolates and phenolic compounds in broccoli. The stressed-broccoli tissue could be subjected to downstream processing in order to extract and purify bioactive molecules with applications in the dietary supplements, agrochemical and cosmetics markets.

  17. 西兰花干燥过程中还原型Vc与氧化型Vc的变化%Changes of ascorbic acid in processes of dehydrated broccoli

    Institute of Scientific and Technical Information of China (English)

    杨霞; 司俊娜; 樊振江; 高愿军

    2012-01-01

    The changes of L-ascorbic acid and I. dehydroascorbic acid during the processes of dehydrated broccoli were studied. The results showed that;the saving of ascorbic acid dealed with steam blanching were higher than treated by hot water, and treaed broccoli 2 mm with hot water in 80℃ had the thinkable result . Broccolis cooling wsth ice-water were better than natural cooling to the saving of ascorbic acid. The ascorbic acid could be saving to the maximize retention treated with vacuum freeze dried, which wass better than treated with microwave drying and hot air drying, and could maintain the shape and the color of broccoli. And treated broccoli 6 h in -25℃ was the best method.%研究西兰花脱水加工过程中还原型Vc和氧化型Vc的变化.结果表明:经过蒸汽热烫的西兰花,其还原型Vc和氧化型Vc的保存率高于热水热烫的保存率,其中80℃蒸汽热烫处理2 min效果最好;热烫后冰水冷却的西兰花还原型Vc和氧化型Vc.保存率高于自然冷却;真空冷冻干燥能将西兰花中还原型Vc和氧化型Vc.最大程度的保留.西兰花脱水加工过程中氧化型Vc的保存率高于还原型Vc,说明西兰花中的还原型Vc更易被破坏.

  18. 聚乳酸薄膜对西兰花的保鲜效果分析%Effect of Polylactic Acid Film Packaging on Preservation of Broccoli

    Institute of Scientific and Technical Information of China (English)

    李伟; 张一珠; 付正义; 张伟阳; 程树军

    2016-01-01

    Polylactic acid (PLA) film was applied to preserve broccoli with polyethylene (PE) film packaging and without packaging as controls. The effect of PLA film on preservation of broccoli quality was investigated by periodically recording quality changes during storage at a constant temperature of (20 ± 3) ℃. The results showed that the vitamin C and chlorophyll contents of broccoli were declined, while the relative conductivity and peroxidase (POD) activity were increased during storage under all three packaging conditions. The broccoli packaged in PLA film showed minimum changes. From the above results it is clear that PLA film can preserve broccoli quality obviously and effectively prolong its shelf life at normal temperature.%用聚乳酸(polylactic acid,PLA)薄膜包装西兰花,以聚乙烯(polyethylene,PE)薄膜包装和无膜包装为对照组,在(20±3)℃条件下对西兰花的相关指标进行定期测定,研究PLA薄膜对西兰花的保鲜效果。结果表明:在PLA薄膜、PE薄膜和无膜包装贮藏过程中,西兰花的VC、叶绿素含量均呈下降趋势,且前者含量明显高于后两者,而相对电导率、过氧化物酶活性均呈上升趋势,前者明显低于后两者。可见,PLA薄膜包装处理对常温条件下的西兰花有明显的保鲜效果,能有效地延长西兰花的常温货架期。

  19. An audit of thiopurine methyltransferase genotyping and phenotyping before intended azathioprine treatment for dermatological conditions

    DEFF Research Database (Denmark)

    Vestergaard, T; Bygum, A

    2009-01-01

    Summary Background. Determining thiopurine methyltransferase (TPMT) genotype and phenotype before azathioprine treatment predicts which patients are most likely to develop myelosuppression. Aim. To evaluate the course of azathioprine treatment in people with TPMT heterozygosity and whether this d...

  20. Arsenic (+3 oxidation state) methyltransferase and the methylation of arsenicals in the invertebrate chordate Ciona intestinalis

    Science.gov (United States)

    Biotransformation of inorganic arsenic (iAs) involves methylation catalyzed by arsenic (+3 oxidation state) methyltransferase (As3mt), yielding mono- , di- , and trimethylated arsenicals. To investigate the evolution of molecular mechanisms that mediate arsenic biotransformation,...

  1. Erythrocuyte catechol-O-methyltransferase activity in related families with schizophrenia

    International Nuclear Information System (INIS)

    Erythrocyte catechol-O-methyltransferase (COMT) activity has been analyzed in 72 individuals from 17 related families with 18 schizophrenic members. No association with any of the three COMT genotypes could be found in this pedigree. (author)

  2. “MGMT for pt mgmt”: Is Methylguanine-DNA Methyltransferase Testing Ready for Patient Management?

    OpenAIRE

    Iafrate, A. John; Louis, David N.

    2008-01-01

    This Commentary reports on a robust quantitative assay for the interpretation of O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation assays that should facilitate the comparison and implementation of such assays across laboratories.

  3. The RNA–Methyltransferase Misu (NSun2) Poises Epidermal Stem Cells to Differentiate

    OpenAIRE

    Sandra Blanco; Agata Kurowski; Jennifer Nichols; Watt, Fiona M.; Salvador Aznar Benitah; Michaela Frye

    2011-01-01

    Homeostasis of most adult tissues is maintained by balancing stem cell self-renewal and differentiation, but whether post-transcriptional mechanisms can regulate this process is unknown. Here, we identify that an RNA methyltransferase (Misu/Nsun2) is required to balance stem cell self-renewal and differentiation in skin. In the epidermis, this methyltransferase is found in a defined sub-population of hair follicle stem cells poised to undergo lineage commitment, and its depletion results in e...

  4. Engineering the DNA cytosine-5 methyltransferase reaction for sequence-specific labeling of DNA

    OpenAIRE

    Lukinavičius, Gražvydas; Lapinaitė, Audronė; Urbanavičiūtė, Giedrė; Gerasimaitė, Rūta; Klimašauskas, Saulius

    2012-01-01

    DNA methyltransferases catalyse the transfer of a methyl group from the ubiquitous cofactor S-adenosyl-L-methionine (AdoMet) onto specific target sites on DNA and play important roles in organisms from bacteria to humans. AdoMet analogs with extended propargylic side chains have been chemically produced for methyltransferase-directed transfer of activated groups (mTAG) onto DNA, although the efficiency of reactions with synthetic analogs remained low. We performed steric engineering of the co...

  5. Structure–function analysis of vaccinia virus mRNA cap (guanine-N7) methyltransferase

    OpenAIRE

    Zheng, Sushuang; Shuman, Stewart

    2008-01-01

    The guanine-N7 methyltransferase domain of vaccinia virus mRNA capping enzyme is a heterodimer composed of a catalytic subunit and a stimulatory subunit. Structure–function analysis of the catalytic subunit by alanine scanning and conservative substitutions (49 mutations at 25 amino acids) identified 12 functional groups essential for methyltransferase activity in vivo, most of which were essential for cap methylation in vitro. Defects in cap binding were demonstrated for a subset of lethal m...

  6. Crystal structure of dengue virus methyltransferase without S-adenosyl-L-methionine.

    Science.gov (United States)

    Noble, Christian G; Li, Shi-Hua; Dong, Hongping; Chew, Sock Hui; Shi, Pei-Yong

    2014-11-01

    Flavivirus methyltransferase is a genetically-validated antiviral target. Crystal structures of almost all available flavivirus methyltransferases contain S-adenosyl-L-methionine (SAM), the methyl donor molecule that co-purifies with the enzymes. This raises a possibility that SAM is an integral structural component required for the folding of dengue virus (DENV) methyltransferase. Here we exclude this possibility by solving the crystal structure of DENV methyltransferase without SAM. The SAM ligand was removed from the enzyme through a urea-mediated denaturation-and-renaturation protocol. The crystal structure of the SAM-depleted enzyme exhibits a vacant SAM-binding pocket, with a conformation identical to that of the SAM-enzyme co-crystal structure. Functionally, equivalent enzymatic activities (N-7 methylation, 2'-O methylation, and GMP-enzyme complex formation) were detected for the SAM-depleted and SAM-containing recombinant proteins. These results clearly indicate that the SAM molecule is not an essential component for the correct folding of DENV methyltransferase. Furthermore, the results imply a potential antiviral approach to search for inhibitors that can bind to the SAM-binding pocket and compete against SAM binding. To demonstrate this potential, we have soaked crystals of DENV methyltransferase without a bound SAM with the natural product Sinefungin and show that preformed crystals are capable of binding ligands in this pocket. PMID:25241250

  7. Effects of broccoli stem and leaf meal on broiler performance, skin pigmentation, antioxidant function, and meat quality.

    Science.gov (United States)

    Hu, C H; Wang, D G; Pan, H Y; Zheng, W B; Zuo, A Y; Liu, J X

    2012-09-01

    Three hundred sixty 1-d-old Ross 308 male broilers were used to study the effects of broccoli stem and leaf meal (BSLM) on growth performance, skin pigmentation, antioxidant function, and meat quality. The chicks were fed 4 diets containing different levels (0, 4.0, 8.0, and 12.0%) of BSLM as partial replacement for corn and soybean meal for a period of 42 d. The results showed that dietary supplementation of BSLM had no effect (P > 0.05) on growth performance. As compared with control, dietary 4%, 8%, and 12% BSLM increased (P poultry products quality with the more skin pigmentation and the less drip loss percentage of breast meat. The more skin pigmentation mainly related to the high amount of xanthophylls in BSLM. The decreased meat drip loss fed BSLM may be caused by the antioxidative function of BSLM. PMID:22912457

  8. Yield and Quality Components of Broccoli Cultivars (Brassica oleracea L. convar. botrytis (L. Alef. var. italica Plenck

    Directory of Open Access Journals (Sweden)

    Nina Toth

    1998-12-01

    According to total yield in the three investigation years it was not possible to point out any cultivar. The highest yield and average weight of top inflorescences was achieved with cultivars Fiesta, Bolivia, Platini and Viking (6.3, 6.2, 6.2 and 6.0 t×ha -1 , resp. 169, 168, 168 and 162 g. Evaluation of the compactness of these inflorescences suggest an exceptional strength. Cultivars Citation and Cruiser excel as regards the production and yield of side inflorescences (2.2 and 2.4 t×ha -1 . For the broccoli production in north-west Croatia during the summer-autumn period recommended cultivars for fresh consumption are Bolivia, Fiesta, Platini and Viking and for processing industries Citation and Cruser.

  9. EFFECT OF SOIL SULFUR FERTILIZER AND SOME FOLIAR FERTILIZERS ON GROWTH AND YIELD OF BROCCOLI IN SALINE SOIL

    Directory of Open Access Journals (Sweden)

    Ali Husain JASIM

    2015-12-01

    Full Text Available Factorial experiment was conducted in the open fields of Agricultural College, Al-Qasim Green University during the agricultural seasons of 2013/2014 and 2014/2015 to study the effect of adding two levels of agricultural sulfur (control and add 100 kg.ha-1 and four levels of nutrient spray (without spray, high-potash fertilizer, high-phosphorus fertilizer and humic acid on growth and yield of broccoli under drip irrigation and polyethylene soil mulching in saline soil (9.6 dS.m-1. Randomized complete block design with three replicates was used. The results showed that agricultural sulfur led to increase number of leaves, leaf area, leaves chlorophyll content, diameter and weight of flower head compared to control. Spraying foliar fertilizer and its interaction with sulfur fertilizer also led to increase all of parameters above (except leaves chlorophyll content significantly compared to control treatment.

  10. Cultivation Technique for Broccoli in Plateau Conditions%西兰花高原栽培技术

    Institute of Scientific and Technical Information of China (English)

    孟平红

    2000-01-01

    @@ 西兰花(broccoli)又名青花菜、绿花菜等,因其色泽碧绿、脆嫩爽口、营养丰富而深受消费者和生产者青睐.但西兰花在我国栽培历史较短,贵州更是栽培较少,远远不能满足市场需求.笔者根据贵州高原的气候特点,经过对西兰花的品种选择、播种期、育苗方法等进行了试验研究,初步摸索出一套适合海拔1 100 m左右的西兰花高原栽培技术.

  11. Aqueous extracts of selenium-fertilized broccoli increase selenoprotein activity and inhibit DNA single-strand breaks, but decrease the activity of quinone reductase in Hepa 1c1c7 cells.

    Science.gov (United States)

    Keck, Anna-Sigrid; Finley, John W

    2006-05-01

    Depending on growth conditions, broccoli may be enriched in the isothiocyanate sulforaphane and/or the mineral selenium (Se); both compounds may play an important role in the reduction of intracellular oxidative stress and chronic disease prevention. Sulforaphane up-regulates transcription of Phase II detoxification proteins (e.g. quinone reductase [QR]), whereas Se is needed for the production of thioredoxin reductase (TR) and glutathione peroxidase-1 (GPx1), both of which exhibit antioxidant activity. The objective of the present study was to determine whether the fertilization of broccoli with Se increases the antioxidant ability of broccoli. Hydrogen peroxide-induced DNA single-strand breaks (measured by single cell electrophoresis, Comet assay) and activity of antioxidant enzymes (GPx, TR and QR) were measured in mouse hepatoma cells (Hepa 1c1c7 cells) treated with purified sulforaphane, sodium selenite or extracts of selenized broccoli. When supplied separately as chemically pure substances, sodium selenite was more effective than sulforaphane for reduction of single-strand breaks. Se-fertilized broccoli extracts were the most effective for reduction of DNA single-strand breaks, and extracts that contained 0.71 microM Se and 0.08 microM sulforaphane inhibited 94% of DNA single-strand breaks. A significant positive association (r = 0.81, p = 0.009) between GPx1 activity and inhibition of DNA single-strand breaks as well as a 24h lag time between addition of Se, sulforaphane or broccoli extract and inhibition of single-strand breaks suggests that some of the antioxidant protection is mediated through selenoproteins. Conversely, fertilization of broccoli with Se decreased the ability of broccoli extract to induce QR activity. These results demonstrate that Se and sulforaphane, alone or as a component of broccoli, may help decrease oxidative stress. They further suggest that Se is the most important for decreasing oxidative stress, but maximizing the Se content

  12. Feasibility study on the use of visible-near-infrared spectroscopy for the screening of individual and total glucosinolate contents in broccoli.

    Science.gov (United States)

    Hernández-Hierro, José Miguel; Valverde, Juan; Villacreces, Salvador; Reilly, Kim; Gaffney, Michael; González-Miret, Maria Lourdes; Heredia, Francisco José; Downey, Gerard

    2012-08-01

    The potential of visible-near-infrared spectroscopy to determine selected individual and total glucosinolates in broccoli has been evaluated. Modified partial least-squares regression was used to develop quantitative models to predict glucosinolate contents. Both the whole spectrum and different spectral regions were separately evaluated to develop the quantitative models; in all cases the best results were obtained using the near-infrared zone between 2000 and 2498 nm. These models have been externally validated for the screening of glucoraphanin, glucobrassicin, 4-methoxyglucobrassicin, neoglucobrassicin, and total glucosinolates contents. In addition, discriminant partial least-squares was used to distinguish between two possible broccoli cultivars and showed a high degree of accuracy. In the case of the qualitative analysis, best results were obtained using the whole spectrum (i.e., 400-2498 nm) with a correct classification rate of 100% in external validation being obtained. PMID:22780224

  13. Effects of Organic and Waste-Derived Fertilizers on Yield, Nitrogen and Glucosinolate Contents, and Sensory Quality of Broccoli (Brassica oleracea L. var. italica).

    Science.gov (United States)

    Øvsthus, Ingunn; Breland, Tor Arvid; Hagen, Sidsel Fiskaa; Brandt, Kirsten; Wold, Anne-Berit; Bengtsson, Gunnar B; Seljåsen, Randi

    2015-12-23

    Organic vegetable production attempts to pursue multiple goals concerning influence on environment, production resources, and human health. In areas with limited availability of animal manure, there is a need for considering various off-farm nutrient resources for such production. Different organic and waste-derived fertilizer materials were used for broccoli production at two latitudes (58° and 67°) in Norway during two years. The fertilizer materials were applied at two rates of total N (80 and 170 kg ha(-1)) and compared with mineral fertilizer (170 kg ha(-1)) and no fertilizer. Broccoli yield was strongly influenced by fertilizer materials (algae meal fertilizer). Yield, but not glucosinolate content, was linearly correlated with estimated potentially plant-available N. However, extruded shrimp shell and mineral NPK fertilizer gave higher glucosinolate contents than sheep manure and no fertilizer. Sensory attributes were less affected by fertilizer material and plant-available N. PMID:26553169

  14. DNA methyltransferase inhibitor CDA-II inhibits myogenic differentiation

    International Nuclear Information System (INIS)

    Highlights: ► CDA-II inhibits myogenic differentiation in a dose-dependent manner. ► CDA-II repressed expression of muscle transcription factors and structural proteins. ► CDA-II inhibited proliferation and migration of C2C12 myoblasts. -- Abstract: CDA-II (cell differentiation agent II), isolated from healthy human urine, is a DNA methyltransferase inhibitor. Previous studies indicated that CDA-II played important roles in the regulation of cell growth and certain differentiation processes. However, it has not been determined whether CDA-II affects skeletal myogenesis. In this study, we investigated effects of CDA-II treatment on skeletal muscle progenitor cell differentiation, migration and proliferation. We found that CDA-II blocked differentiation of murine myoblasts C2C12 in a dose-dependent manner. CDA-II repressed expression of muscle transcription factors, such as Myogenin and Mef2c, and structural proteins, such as myosin heavy chain (Myh3), light chain (Mylpf) and MCK. Moreover, CDA-II inhibited C1C12 cell migration and proliferation. Thus, our data provide the first evidence that CDA-II inhibits growth and differentiation of muscle progenitor cells, suggesting that the use of CDA-II might affect skeletal muscle functions.

  15. Theoretical insights into catalytic mechanism of protein arginine methyltransferase 1.

    Directory of Open Access Journals (Sweden)

    Ruihan Zhang

    Full Text Available Protein arginine methyltransferase 1 (PRMT1, the major arginine asymmetric dimethylation enzyme in mammals, is emerging as a potential drug target for cancer and cardiovascular disease. Understanding the catalytic mechanism of PRMT1 will facilitate inhibitor design. However, detailed mechanisms of the methyl transfer process and substrate deprotonation of PRMT1 remain unclear. In this study, we present a theoretical study on PRMT1 catalyzed arginine dimethylation by employing molecular dynamics (MD simulation and quantum mechanics/molecular mechanics (QM/MM calculation. Ternary complex models, composed of PRMT1, peptide substrate, and S-adenosyl-methionine (AdoMet as cofactor, were constructed and verified by 30-ns MD simulation. The snapshots selected from the MD trajectory were applied for the QM/MM calculation. The typical SN2-favored transition states of the first and second methyl transfers were identified from the potential energy profile. Deprotonation of substrate arginine occurs immediately after methyl transfer, and the carboxylate group of E144 acts as proton acceptor. Furthermore, natural bond orbital analysis and electrostatic potential calculation showed that E144 facilitates the charge redistribution during the reaction and reduces the energy barrier. In this study, we propose the detailed mechanism of PRMT1-catalyzed asymmetric dimethylation, which increases insight on the small-molecule effectors design, and enables further investigations into the physiological function of this family.

  16. Characterization of the phospholipid methyltransferase in RBC ghost preparations

    International Nuclear Information System (INIS)

    The activity of the phospholipid methyltransferase from human RBC ghosts was studied using radio-HPLC techniques to analyze the products. Both monomethyl phosphatidyl ethanolamine (MMPE) and dimethyl phosphatidyl ethanolamine (DMPE) were used as substrated. The reaction rate was linear for 45 min. Apparent K/sub M/s of 24-28 uM and 19-21 uM were measured for these two substrates, respectively. The reaction rate was not linear with protein. It appeared to increase logarithmic. An apparent K/sub M/ for S-adenosylmethionine was 36-45 uM. These K/sub M/ values are similar to those reported by others for liver. As the concentration of MMPE was increased, the ratio of DMPE/PC also increased due largely to a greater increase in DMPE formation. Optimal reaction rates for the formation of DMPE were 0.9-1.3 pmol/mg/min, and an optimal rate of about 1.7-2.4 pmol/min/mg was measured for the conversion of DMPE to phosphatidyl choline (PC). Freezing the ghost preparation did not affect the activity of the enzyme. When no exogenous phospholipid was added to the incubation, the sum of the formation rates of all three methylated products was about 26 pmol/mg/hr. The relative amount of each product was 46% MMPE, 32% DMPE and 22% PC. When either MMPE or DMPE was added as substrate, the formation of MMPE was reduced to less than 1%

  17. Thiopurine methyltransferase activity in red blood cells of dogs.

    Science.gov (United States)

    Kidd, Linda Benjamin; Salavaggione, Oreste E; Szumlanski, Carol L; Miller, Jackie L; Weinshilboum, Richard M; Trepanier, Lauren

    2004-01-01

    Thiopurine methyltransferase (TPMT) is an important enzyme in the metabolism of thiopurine medications such as azathioprine. In humans, activity varies widely among individuals, primarily because of genetic polymorphisms. Low TPMT activity increases the risk of myelosuppression from azathioprine and 6-mercaptopurine, whereas high TPMT activity is associated with poor drug efficacy. The purpose of this study was to determine whether dogs also show a wide range of TPMT activity. Heparinized blood samples were obtained from 177 dogs associated with a veterinary teaching hospital. Red blood cell (RBC) TPMT activity was measured by means of a modification of a radiochemical method as established for use in people. TPMT activity varied across a 9-fold range (7.9-71.8 U of RBC per milliliter; median, 21.7). Variation in TPMT activity was not associated with age, sex, or neutering status. Giant Schnauzers had much lower TPMT activity (7.9-20 U of RBC per milliliter; median, 13.1; P dogs could affect thiopurine drug toxicity and efficacy in canine patients. PMID:15058773

  18. Human erythrocyte thiol methyltransferase: radiochemical microassay and biochemical properties

    International Nuclear Information System (INIS)

    A radiochemical microassay for the measurement of thiol methyltransferase (TMT) activity in human red blood cell (RBC) membranes has been developed. Both 2-mercaptoethanol and dithiothreitol were used as substrates for the enzyme. The pH optimum of the reaction was approximately 9.0 when glycine-NaOH was used as a buffer. The apparent Michaelis-Menten (Ksub(M)) value for the methyl donor for the reaction, S-adenosyl-L-methionine, was 43 μmol/l. Human RBC TMT activity was neither activated nor inhibited by Ca2+, Mg2+, or tropolone, but the enzyme was inhibited by SKF 525A and by reagents that react with sulfhydryl grcups. The mean TMT activity in blood from 289 randomly selected adult white subjects was 10.93 +- 3.22 units per mg protein (mean +- S.D.). The activity was the same in samples from men and women. The results of experiments in which TMT activity was measured in mixtures of RBC membranes with relatively ''low'' and relatively ''high'' activities provided no evidence that individual variations in the enzyme activity were due to variations in endogenous TMT activators or inhibitors. (Auth.)

  19. Identification and functional characterization of lysine methyltransferases of Entamoeba histolytica.

    Science.gov (United States)

    Borbolla-Vázquez, Jessica; Orozco, Esther; Medina-Gómez, Christian; Martínez-Higuera, Aarón; Javier-Reyna, Rosario; Chávez, Bibiana; Betanzos, Abigail; Rodríguez, Mario A

    2016-07-01

    Lysine methylation of histones, a posttranslational modification catalyzed by lysine methyltransferases (HKMTs), plays an important role in the epigenetic regulation of transcription. Lysine methylation of non-histone proteins also impacts the biological function of proteins. Previously it has been shown that lysine methylation of histones of Entamoeba histolytica, the protozoan parasite that infects 50 million people worldwide each year and causing up to 100,000 deaths annually, is implicated in the epigenetic machinery of this microorganism. However, the identification and characterization of HKMTs in this parasite had not yet been determined. In this work we identified four HKMTs in E. histolytica (EhHKMT1 to EhHKMT4) that are expressed by trophozoites. Enzymatic assays indicated that all of them are able to transfer methyl groups to commercial histones. EhHKMT1, EhHKMT2 and EhHKMT4 were detected in nucleus and cytoplasm of trophozoites. In addition EhHKMT2 and EhHKMT4 were located in vesicles containing ingested cells during phagocytosis, and they co-immunoprecipitated with EhADH, a protein involved in the phagocytosis of this parasite. Results suggest that E. histolytica uses its HKMTs to regulate transcription by epigenetic mechanisms, and at least two of them could also be implicated in methylation of proteins that participate in phagocytosis. PMID:27062489

  20. DNA methyltransferase inhibitor CDA-II inhibits myogenic differentiation

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Zirong [State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou 510060 (China); Department of Molecular Genetics and Microbiology, Shands Cancer Center, University of Florida, Gainesville, FL 32610 (United States); Jin, Guorong [State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou 510060 (China); Lin, Shuibin [Department of Molecular Genetics and Microbiology, Shands Cancer Center, University of Florida, Gainesville, FL 32610 (United States); Lin, Xiumei [Department of Hematology, Guangzhou First Municipal People' s Hospital, Guangzhou 510180 (China); Gu, Yumei [Department of Molecular Genetics and Microbiology, Shands Cancer Center, University of Florida, Gainesville, FL 32610 (United States); Zhu, Yujuan; Hu, Chengbin; Zhang, Qingjiong [State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou 510060 (China); Wu, Lizi [Department of Molecular Genetics and Microbiology, Shands Cancer Center, University of Florida, Gainesville, FL 32610 (United States); Shen, Huangxuan, E-mail: shenhx@mail.sysu.edu.cn [State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou 510060 (China)

    2012-06-08

    Highlights: Black-Right-Pointing-Pointer CDA-II inhibits myogenic differentiation in a dose-dependent manner. Black-Right-Pointing-Pointer CDA-II repressed expression of muscle transcription factors and structural proteins. Black-Right-Pointing-Pointer CDA-II inhibited proliferation and migration of C2C12 myoblasts. -- Abstract: CDA-II (cell differentiation agent II), isolated from healthy human urine, is a DNA methyltransferase inhibitor. Previous studies indicated that CDA-II played important roles in the regulation of cell growth and certain differentiation processes. However, it has not been determined whether CDA-II affects skeletal myogenesis. In this study, we investigated effects of CDA-II treatment on skeletal muscle progenitor cell differentiation, migration and proliferation. We found that CDA-II blocked differentiation of murine myoblasts C2C12 in a dose-dependent manner. CDA-II repressed expression of muscle transcription factors, such as Myogenin and Mef2c, and structural proteins, such as myosin heavy chain (Myh3), light chain (Mylpf) and MCK. Moreover, CDA-II inhibited C1C12 cell migration and proliferation. Thus, our data provide the first evidence that CDA-II inhibits growth and differentiation of muscle progenitor cells, suggesting that the use of CDA-II might affect skeletal muscle functions.

  1. Sensitivity of produce respiration models used in the MAP-DESIGN software on the shelf life simulation of broccoli in the modified atmosphere package

    OpenAIRE

    Weerasak Lertsiriyothin

    2009-01-01

    Optimization for the designing of modified atmosphere packaging (MAP) for broccoli was made by the MAP-DESIGN software (author’s own code). The software is capable of dealing with all parameters required for the designing of MAP for fresh produce, namely packaging materials, package dimensions, storage conditions, and plant respiratory models. Computational algorithms were carefully designed based upon widely-used theories of living plant respiration, gas permeability through packaging film...

  2. Effect of devernalization on the transition from vegetative to prefloral phase of the broccoli (Brassica oleracea var. italica cv. 'Fiesta' shoot meristem

    Directory of Open Access Journals (Sweden)

    Elżbieta Zenkteler

    2012-12-01

    Full Text Available The morphological and histological changes which occur in the cold-induced apical meristem of broccoli (B. oleracea var. italica cv. ‘Fiesta’ during transition from the vegetative to the generative phase have been demonstrated in our previous investigations. Now, a light microscopic study on the micromorphological parameters of the tunica and corpus of the meristem was undertaken to ascertain their association with devernalisation. Changes in the apex of broccoli seedlings grown for 26 days at 24oC, succeeded by 35 days of cold treatment (16oC - vernalization followed by 14 days of warm treatment (24oC - devernalization, were studied and described. Microscopic analysis according to the new method of slide preparation showed cytohistological zonation and the spatial distribution of cells in the central and peripheral zones of the broccoli shoot meristem more precisely than possible hitherto. Comparison between subsequent stages of meristem reorganization after devernalization revealed variation in the structure of the apex during its transition to the evocation phase of flowering. The results of measurements show that the tunica width changes irregularly, increasing from its minimum value (112.39±5.78 μm through the medium one to the maximum values (260.32±2.50, to decrease again, while its height increased and decreased sinusoidally from its minimum value (21.52±4.30μm to the maximum one (76.98±7.44μm and then decreasing again to the minimum value (21.52±4.30. The stratification of the tunica as well as its width and height revealed relatively low variation within the analyzed time. These structural traits correlated with cold/warm treatments may be further utilized in the broccoli breeding programme against developmental disturbances.

  3. Effects of the 3D-clinorotation on endogenous substances of broccoli sprout (Brassica oleracea var. italica) and its food safety

    Science.gov (United States)

    Hiraishi, K.; Tomita-Yokotani, K.; Wakabayashi, K.; Hashimoto, H.; Miyagawa, T.; Yamashita, M.

    Habitation in outer space is one of our challenges in this century We are studying on space agriculture to provide foods for space living people However careful assessment should be made on the effects of exotic environment on the endogenous production of biologically active substances and food safety of plants cultivated in space Broccoli sprout Brassica oleracea var italica is known to produce sulforaphane 4-methylsulfinybutyl isothiocyanate which is effective to function as an antioxidant and enhance immunity Because of such substance it is recognized to be good food materials Broccoli sprouts were then cultivated for 3 days under the 3D-clinorotation The amount of sulforaphane produced by this treatment showed no significant difference compared to the ground control Secondly we examined population of microorganisms adhered on the surface of sprout cultivated under the 3D-clinorotation Number of the microorganisms colony formed was statistically higher than the control Mold species was identified to penicillium sp based on the microscopic observation Poor construction of plant cell wall elements cellulose lignin etc is well known effects of microgravity Defense function of the broccoli plant cells might be weakened against microorganism We also speculate other possible causes for the high rate of contamination such as photosynthetic activity of the plant or microclimate air flow heat transport and humidity around the seedling affected by pseudo-microgravity or the 3D-clinorotation Those factors may relate to the difference in proliferation

  4. Change of related isoenzyme during vernalization in broccoli%青花菜绿体春化过程中同工酶变化研究

    Institute of Scientific and Technical Information of China (English)

    刘生财; 蒋欣梅; 于锡宏

    2009-01-01

    试验以青花菜早熟品种"清风103"为试材,对青花菜绿体春化过程中植株中春化相关的酶进行研究.结果表明,青花菜植株在低温春化处理过程中,NR、EST和α-淀粉酶及ASP的活性都呈增加趋势.对EST、α-淀粉酶和POD同工酶分析发现,经过低温春化处理后既有新谱带的产生,也有谱带的消失,产生或消失的谱带可能与青花菜春化有关.%The experiment took early maturing variety broccoli Qingfeng 103 as experimental material. Related isoenzyme during vernalization in broccoli was studied in this research. The results showed that NR, EST, α-amylase and ASP activity were significantly increased during vernalization. After the analyse of EST, α-amylase and POD isoenzyme, some new bands appeared but some disappeared. Perhaps these bands were associated with vernalization in broccoli.

  5. Assessing Competence of Broccoli Consumption on Inflammatory and Antioxidant Pathways in Restraint-Induced Models: Estimation in Rat Hippocampus and Prefrontal Cortex

    Directory of Open Access Journals (Sweden)

    Leila Khalaj

    2013-01-01

    Full Text Available A growing body of evidence advocated the protective and therapeutic potential of natural compounds and phytochemicals used in diets against pathological conditions. Herein, the outcome of dietary whole broccoli consumption prior to restraint stress has been investigated in the hippocampus and prefrontal cortex of male rats, two important regions involved in the processing of responses to stressful events. Interestingly, a region-specific effect was detected regarding some of antioxidant defense system factors: nuclear factor erythroid-derived 2-related factor 2 (Nrf-2 antioxidant pathway, mitochondrial prosurvival proteins involved in mitochondrial biogenesis, and apoptotic cell death proteins. Dietary broccoli supplementation modulated the restraint-induced changes towards a consistent overall protection in the hippocampus. In the prefrontal cortex, however, despite activation of most of the protective factors, presumably as an attempt to save the system against the stress insult, some detrimental outcomes such as induced malate dehydrogenase (MDA level and cleaved form of caspase-3 were detectable. Such diversity may be attributed in one hand to the different basic levels and/or availability of defensive mechanisms within the two studied cerebral regions, and on the other hand to the probable dose-dependent and hormetic effects of whole broccoli. More experiments are essential to demonstrate these assumptions.

  6. 不同贮藏条件对青花菜品质的影响%Effects of Storage Conditions on the Quality of Broccolis

    Institute of Scientific and Technical Information of China (English)

    杨佩; 侯军; 石英

    2011-01-01

    Effects of such conditions as temperature, relative humidity and light on storage quality of broccoli were studied. The results showed that, in the three combinations of temperature and relative humidity, the weight and L-ascorbic acid of broccoli and hardness of stem could be maintained by the condition of 15 ℃ and 75% relative humidity. Under the lighting conditions (at the temperature of 20±2 ℃ ), the soluble solids contents (SSC) of buds and the weight of broccoli were decreased slowly. But L-ascorbic acid and hardness decreased more quickly under the lighting conditions.%研究了不同温湿度及光照条件对青花菜采后贮藏品质的影响.结果表明,在三种温湿度组合处理中,以15℃温度、75%相对湿度的条件可有效减少青花菜的失重,保持花茎的硬度和还原型抗坏血酸含量,延长贮藏期;室温(20±2℃)光照条件有利于减少青花菜的失重和可溶性固形物含量的下降速率,但不利于硬度及还原型抗坏血酸的保持.

  7. 青花菜气相色谱指纹图谱研究%Study on Gas Chromatographic Fingerprint of Broccoli

    Institute of Scientific and Technical Information of China (English)

    王冬群; 胡寅侠

    2014-01-01

    With FPD as detector , the fingerprint features of broccoli were studied by gas chromatographic method using DB-1701 and HP-5 capillary column respectively .The results showed that the broccoli had 2 peaks in both 2 columns, and their retention times were all very stable .The peak areas among different sam-ples were obviously different , and there was no obvious proportional relation between 2 peaks of the same sam-ple.The GC fingerprint could be used as one of the bases of broccoli quality assessment .%采用气相色谱法,分别利用DB-1701和HP-5毛细管柱,以FPD为检测器,研究了青花菜的指纹图谱特点。结果显示,青花菜在2种柱子中均表现为2个峰,且其保留时间均非常稳定,但不同样品间峰面积差异较大;同一样品中2个峰的峰面积没有明显的比例关系。气相色谱指纹图谱可作为青花菜质量控制的依据之一。

  8. 西兰花甜玉米复合蔬菜纸的研制%Study on Compound Vegetable Paper of Broccoli and Sweet Corn

    Institute of Scientific and Technical Information of China (English)

    汤慧民; 尚海英

    2015-01-01

    With broccoli and sweet corn as raw material, sodium alginate, carrageenan,starch as adhesive, based on single factor experiment, the production process conditions of vegetable paper of broccoli and sweet corn was optimized through orthogonal experiment.The result showed that the acquired vegetable paper was best in color, flavor, taste and texture adopting the condition of the quality ratio of broccoli slurry and sweet corn pulp was 2:1,sodium alginate 0.5%, carrageenan 0.6%, starch 8%, film thickness 1.5 mm, drying 4 h at 65℃.%以西兰花和甜玉米为原料,海藻酸钠、卡拉胶、淀粉为胶黏剂,在单因素的基础上,采用正交试验法确定西兰花甜玉米复合蔬菜纸的最佳生产工艺条件:西兰花浆和甜玉米浆的质量比例是2:1、0.5%的海藻酸钠、0.6%卡拉胶、8%淀粉,涂膜厚度1.5 mm,65℃干燥4 h.所制得的蔬菜纸色、香、味、形俱佳.

  9. A Picrinine N-Methyltransferase Belongs to a New Family of γ-Tocopherol-Like Methyltransferases Found in Medicinal Plants That Make Biologically Active Monoterpenoid Indole Alkaloids.

    Science.gov (United States)

    Levac, Dylan; Cázares, Paulo; Yu, Fang; De Luca, Vincenzo

    2016-04-01

    Members of the Apocynaceae plant family produce a large number of monoterpenoid indole alkaloids (MIAs) with different substitution patterns that are responsible for their various biological activities. A novel N-methyltransferase involved in the vindoline pathway in Catharanthus roseus showing distinct similarity to γ-tocopherol C-methyltransferases was used in a bioinformatic screen of transcriptomes from Vinca minor, Rauvolfia serpentina, and C. roseus to identify 10 γ-tocopherol-like N-methyltransferases from a large annotated transcriptome database of different MIA-producing plant species (www.phytometasyn.ca). The biochemical function of two members of this group cloned from V. minor (VmPiNMT) and R. serpentina (RsPiNMT) have been characterized by screening their biochemical activities against potential MIA substrates harvested from the leaf surfaces of MIA-accumulating plants. The approach was validated by identifying the MIA picrinine from leaf surfaces of Amsonia hubrichtii as a substrate of VmPiNMT and RsPiNMT. Recombinant proteins were shown to have high substrate specificity and affinity for picrinine, converting it to N-methylpicrinine (ervincine). Developmental studies with V. minor and R. serpentina showed that RsPiNMT and VmPiNMT gene expression and biochemical activities were highest in younger leaf tissues. The assembly of at least 150 known N-methylated MIAs within members of the Apocynaceae family may have occurred as a result of the evolution of the γ-tocopherol-like N-methyltransferase family from γ-tocopherol methyltransferases. PMID:26848097

  10. An enzyme-coupled continuous spectrophotometric assay for S-adenosylmethionine-dependent methyltransferases.

    Science.gov (United States)

    Dorgan, Kathleen M; Wooderchak, Whitney L; Wynn, Donraphael P; Karschner, Erin L; Alfaro, Joshua F; Cui, Yinqiu; Zhou, Zhaohui Sunny; Hevel, Joan M

    2006-03-15

    Modification of small molecules and proteins by methyltransferases affects a wide range of biological processes. Here, we report an enzyme-coupled continuous spectrophotometric assay to quantitatively characterize S-adenosyl-L-methionine (AdoMet/SAM)-dependent methyltransferase activity. In this assay, S-adenosyl-L-homocysteine (AdoHcy/SAH), the transmethylation product of AdoMet-dependent methyltransferases, is hydrolyzed to S-ribosylhomocysteine and adenine by recombinant S-adenosylhomocysteine/5'-methylthioadenosine nucleosidase (SAHN/MTAN, EC 3.2.2.9). Subsequently, adenine generated from AdoHcy is further hydrolyzed to hypoxanthine and ammonia by recombinant adenine deaminase (EC 3.5.4.2). This deamination is associated with a decrease in absorbance at 265 nm that can be monitored continuously. Coupling enzymes are recombinant and easily purified. The utility of this assay was shown using recombinant rat protein arginine N-methyltransferase 1 (PRMT1, EC 2.1.1.125), which catalyzes the mono- and dimethylation of guanidino nitrogens of arginine residues in select proteins. Using this assay, the kinetic parameters of PRMT1 with three synthetic peptides were determined. An advantage of this assay is the destruction of AdoHcy by AdoHcy nucleosidase, which alleviates AdoHcy product feedback inhibition of S-adenosylmethionine-dependent methyltransferases. Finally, this method may be used to assay other enzymes that produce AdoHcy, 5'-methylthioadenosine, or compounds that can be cleaved by AdoHcy nucleosidase. PMID:16460659

  11. Conservation and Functional Importance of Carbon-Oxygen Hydrogen Bonding in AdoMet-Dependent Methyltransferases

    Energy Technology Data Exchange (ETDEWEB)

    Horowitz, Scott; Dirk, Lynnette M.A.; Yesselman, Joseph D.; Nimtz, Jennifer S.; Adhikari, Upendra; Mehl, Ryan A.; Scheiner, Steve; Houtz, Robert L.; Al-Hashimi, Hashim M.; Trievel, Raymond C. [Oregon State U.; (Michigan); (Utah SU); (HHMI); (Kentucky)

    2013-09-06

    S-Adenosylmethionine (AdoMet)-based methylation is integral to metabolism and signaling. AdoMet-dependent methyltransferases belong to multiple distinct classes and share a catalytic mechanism that arose through convergent evolution; however, fundamental determinants underlying this shared methyl transfer mechanism remain undefined. A survey of high-resolution crystal structures reveals that unconventional carbon–oxygen (CH···O) hydrogen bonds coordinate the AdoMet methyl group in different methyltransferases irrespective of their class, active site structure, or cofactor binding conformation. Corroborating these observations, quantum chemistry calculations demonstrate that these charged interactions formed by the AdoMet sulfonium cation are stronger than typical CH···O hydrogen bonds. Biochemical and structural studies using a model lysine methyltransferase and an active site mutant that abolishes CH···O hydrogen bonding to AdoMet illustrate that these interactions are important for high-affinity AdoMet binding and transition-state stabilization. Further, crystallographic and NMR dynamics experiments of the wild-type enzyme demonstrate that the CH···O hydrogen bonds constrain the motion of the AdoMet methyl group, potentially facilitating its alignment during catalysis. Collectively, the experimental findings with the model methyltransferase and structural survey imply that methyl CH···O hydrogen bonding represents a convergent evolutionary feature of AdoMet-dependent methyltransferases, mediating a universal mechanism for methyl transfer.

  12. Impact of single-gene and dual-gene Bt broccoli on the herbivore Pieris rapae (Lepidoptera: Pieridae) and its pupal endoparasitoid Pteromalus puparum (Hymenoptera: Pteromalidae).

    Science.gov (United States)

    Chen, Mao; Zhao, Jian-zhou; Shelton, Anthony M; Cao, Jun; Earle, Elizabeth D

    2008-08-01

    Transgenic brassica crops producing insecticidal proteins from Bacillus thuringiensis (Bt) are being investigated as candidates for field release to control lepidopteran pests. Information on the potential impact of Bt brassica crops on pests and non-target natural enemies is needed as part of an environmental risk assessment prior to the commercial release. This first tier study provides insight into the tritrophic interactions among Bt broccoli plants, the herbivore Pieris rapae and its parasitoid Pteromalus puparum. We first evaluated the efficacy of three types of Bt broccoli plants, cry1Ac, cry1C and cry1Ac + cry1C, on different instars of P. rapae. Bt broccoli effectively controlled P. rapae larvae, although later instars were more tolerant. The efficacy of different Bt broccoli plants on P. rapae larvae was consistently cry1Ac > cry1Ac + cry1C > cry1C. When the parasitoid P. puparum developed in a P. rapae pupa (host) that had developed from Bt plant-fed older larvae, developmental time, total number and longevity of the P. puparum generated from the Bt plant-fed host were significantly affected compared with those generated from the non-Bt control plant-fed host. Simultaneously, negative effects on P. rapae pupae were found, i.e. pupal length, width and weight were significantly reduced after older P. rapae larvae fed on different Bt plants for 1 or 2 days. Cry1C toxin was detected using ELISA in P. rapae pupae after older larvae fed on cry1C broccoli. However, no Cry1C toxin was detected in newly emerged P. puparum adults developing in Bt-fed hosts. Only a trace amount of toxin was detected from entire P. puparum pupae dissected from the Bt plant-fed host. Moreover, no negative effect was found on the progeny of P. puparum developing from the Bt plant-fed host when subsequently supplied with a healthy host, P. rapae pupae. The reduced quality of the host appears to be the only reason for the observed deleterious effects on P. puparum. Our data suggest that

  13. Research of Natamycin on the Freshness Retaining of Broccolis%纳他霉素对西兰花的保鲜效果

    Institute of Scientific and Technical Information of China (English)

    林本芳; 鲁晓翔; 李江阔; 陈绍慧

    2012-01-01

    研究纳他霉素处理对西兰花冷藏保鲜品质的影响。纳他霉素处理浓度分别为800mg/L、1200mg/L,在(0±1)℃冷藏期间每10d取样测定西兰花理化指标及营养成分。结果表明:与冷藏对照相比,纳他霉素处理对于延缓西兰花可溶性固形物含量、Vc含量及叶绿素含量的下降均有明显效果,并可抑制西兰花贮藏期间的呼吸强度;纳他霉素处理可提高西兰花SOD的活性和降低PPO活性,延缓了细胞膜相对透性的升高。说明冷藏结合纳他霉素处理对西兰花的成熟衰老有抑制作用,其中经1200mg/L处理的西兰花保鲜效果较好。%This paper investigated the effect of different concentrations (800mg/L,1200mg/L) of natamycin on the quality of Broccolis stored at -1 ℃ - 1 ℃. The physicochemical indexes and nutrition components of Broccolis were detected every 10days. The results showed that the soluble solids, vitamine C and chlorophyll content of broccolis by natamycin treatment decreased less than that just by cold storage. Natamycin could inhibit the increase of respiratory intensity, increased superoxide dismutase (SOD) activities and slowed down polyphenol oxidase(PPO) activity and de- creased cell membrane permeability. These results showed that cold storage combined with natamycin treatment could delay ripening of Broccoli, wherein the storage effect of Broccolis with 1200 mg/L natamycin treatment was better than that with 800mg/L.

  14. Methyltransferases mediate cell memory of a genotoxic insult.

    Science.gov (United States)

    Rugo, R E; Mutamba, J T; Mohan, K N; Yee, T; Chaillet, J R; Greenberger, J S; Engelward, B P

    2011-02-10

    Characterization of the direct effects of DNA-damaging agents shows how DNA lesions lead to specific mutations. Yet, serum from Hiroshima survivors, Chernobyl liquidators and radiotherapy patients can induce a clastogenic effect on naive cells, showing indirect induction of genomic instability that persists years after exposure. Such indirect effects are not restricted to ionizing radiation, as chemical genotoxins also induce heritable and transmissible genomic instability phenotypes. Although such indirect induction of genomic instability is well described, the underlying mechanism has remained enigmatic. Here, we show that mouse embryonic stem cells exposed to γ-radiation bear the effects of the insult for weeks. Specifically, conditioned media from the progeny of exposed cells can induce DNA damage and homologous recombination in naive cells. Notably, cells exposed to conditioned media also elicit a genome-destabilizing effect on their neighbouring cells, thus demonstrating transmission of genomic instability. Moreover, we show that the underlying basis for the memory of an insult is completely dependent on two of the major DNA cytosine methyltransferases, Dnmt1 and Dnmt3a. Targeted disruption of these genes in exposed cells completely eliminates transmission of genomic instability. Furthermore, transient inactivation of Dnmt1, using a tet-suppressible allele, clears the memory of the insult, thus protecting neighbouring cells from indirect induction of genomic instability. We have thus demonstrated that a single exposure can lead to long-term, genome-destabilizing effects that spread from cell to cell, and we provide a specific molecular mechanism for these persistent bystander effects. Collectively, our results impact the current understanding of risks from toxin exposures and suggest modes of intervention for suppressing genomic instability in people exposed to carcinogenic genotoxins. PMID:21057543

  15. Catechol-O-methyltransferase, dopamine, and sleep-wake regulation.

    Science.gov (United States)

    Dauvilliers, Yves; Tafti, Mehdi; Landolt, Hans Peter

    2015-08-01

    Sleep and sleep disorders are complex and highly variable phenotypes regulated by many genes and environment. The catechol-O-methyltransferase (COMT) gene is an interesting candidate, being one of the major mammalian enzymes involved in the catabolism of catecholamines. The activity of COMT enzyme is genetically polymorphic due to a guanine-to-adenine transition at codon 158, resulting in a valine (Val) to methionine (Met) substitution. Individuals homozygous for the Val allele show higher COMT activity, and lower dopaminergic signaling in prefrontal cortex (PFC) than subjects homozygous for the Met allele. Since COMT has a crucial role in metabolising dopamine, it was suggested that the common functional polymorphism in the COMT gene impacts on cognitive function related to PFC, sleep-wake regulation, and potentially on sleep pathologies. The COMT Val158Met polymorphism may predict inter-individual differences in brain electroencephalography (EEG) alpha oscillations and recovery processes resulting from partial sleep loss in healthy individuals. The Val158Met polymorphism also exerts a sexual dimorphism and has a strong effect on objective daytime sleepiness in patients with narcolepsy-cataplexy. Since the COMT enzyme inactivates catecholamines, it was hypothesized that the response to stimulant drugs differs between COMT genotypes. Modafinil maintained executive functioning performance and vigilant attention throughout sleep deprivation in subjects with Val/Val genotype, but less in those with Met/Met genotype. Also, homozygous Met/Met patients with narcolepsy responded to lower doses of modafinil compared to Val/Val carriers. We review here the critical role of the common functional COMT gene polymorphism, COMT enzyme activity, and the prefrontal dopamine levels in the regulation of sleep and wakefulness in normal subjects, in narcolepsy and other sleep-related disorders, and its impact on the response to psychostimulants. PMID:25466290

  16. Demonstration of thiopurine methyltransferase activity in the erythrocytes of cats.

    Science.gov (United States)

    Foster, A P; Shaw, S E; Duley, J A; Shobowale-Bakre, E M; Harbour, D A

    2000-01-01

    Azathioprine is a purine analogue used as an immunosuppressive and immunomodulator agent in various mammals, including cats. Several adverse reactions have been reported and have limited the use of the drug in the cat. Adverse reactions to azathioprine in humans have been correlated with reduced activity of thiopurine methyltransferase (TPMT) in erythrocytes. The purpose of this preliminary study was to determine if cats have TPMT activity in their erythrocytes and to compare the values obtained with the normal range for humans and the normal range for dogs in a preliminary report. Activity of the enzyme was measured in blood samples drawn from 41 cats. Blood also was taken from 5 dogs. The mean erythrocyte TPMT activity in the cats was 2.4 +/- 0.4 nmol (range, 1.2-3.9 nmol) per hour per milliliter of red blood cells (U/mL RBC) or 2-8 nmol per hour per gram of hemoglobin (U/g Hb). This range was far lower than the normal human range (8-15 U/mL RBC; 16-33 U/g Hb) and was of monopolar distribution. This observation apparently precludes any diagnostic purpose in assaying erythrocyte TPMT in this species. Erythrocyte TPMT activity in the 5 dogs ranged from 5.5 to 13.1 U/mL RBC (11-27 U/g Hb), which was comparable with normal and carrier ranges for humans, but proof of TPMT genetic polymorphism in either species will require genotyping studies. PMID:11012121

  17. Two distinct arginine methyltransferases are required for biogenesis of Sm-class ribonucleoproteins

    Science.gov (United States)

    Gonsalvez, Graydon B.; Tian, Liping; Ospina, Jason K.; Boisvert, François-Michel; Lamond, Angus I.; Matera, A. Gregory

    2007-01-01

    Small nuclear ribonucleoproteins (snRNPs) are core components of the spliceosome. The U1, U2, U4, and U5 snRNPs each contain a common set of seven Sm proteins. Three of these Sm proteins are posttranslationally modified to contain symmetric dimethylarginine (sDMA) residues within their C-terminal tails. However, the precise function of this modification in the snRNP biogenesis pathway is unclear. Several lines of evidence suggest that the methyltransferase protein arginine methyltransferase 5 (PRMT5) is responsible for sDMA modification of Sm proteins. We found that in human cells, PRMT5 and a newly discovered type II methyltransferase, PRMT7, are each required for Sm protein sDMA modification. Furthermore, we show that the two enzymes function nonredundantly in Sm protein methylation. Lastly, we provide in vivo evidence demonstrating that Sm protein sDMA modification is required for snRNP biogenesis in human cells. PMID:17709427

  18. The RNA-methyltransferase Misu (NSun2 poises epidermal stem cells to differentiate.

    Directory of Open Access Journals (Sweden)

    Sandra Blanco

    2011-12-01

    Full Text Available Homeostasis of most adult tissues is maintained by balancing stem cell self-renewal and differentiation, but whether post-transcriptional mechanisms can regulate this process is unknown. Here, we identify that an RNA methyltransferase (Misu/Nsun2 is required to balance stem cell self-renewal and differentiation in skin. In the epidermis, this methyltransferase is found in a defined sub-population of hair follicle stem cells poised to undergo lineage commitment, and its depletion results in enhanced quiescence and aberrant stem cell differentiation. Our results reveal that post-transcriptional RNA methylation can play a previously unappreciated role in controlling stem cell fate.

  19. H3K9 methyltransferase G9a and the related molecule GLP

    OpenAIRE

    Shinkai, Yoichi; Tachibana, Makoto

    2011-01-01

    The discovery of Suv39h1, the first SET domain-containing histone lysine methyltransferase (HKMT), was reported in 2000. Since then, research on histone methylation has progressed rapidly. Among the identified HKMTs in mammals, G9a and GLP are the primary enzymes for mono- and dimethylation at Lys 9 of histone H3 (H3K9me1 and H3K9me2), and exist predominantly as a G9a–GLP heteromeric complex that appears to be a functional H3K9 methyltransferase in vivo. Recently, many important studies have ...

  20. Novel Function of Lysine Methyltransferase G9a in the Regulation of Sox2 Protein Stability

    OpenAIRE

    Lee, Jae-Young; Lee, Se-Hwan; Heo, Sun-Hee; Kim, Kwang-Soo; Kim, Changhoon; Kim, Dae-Kwan; Ko, Jeong-Jae; Park, Kyung-Soon

    2015-01-01

    G9a is a lysine methyltransferase (KMTase) for histone H3 lysine 9 that plays critical roles in a number of biological processes. Emerging evidence suggests that aberrant expression of G9a contributes to tumor metastasis and maintenance of a malignant phenotype in cancer by inducing epigenetic silencing of tumor suppressor genes. Here, we show that G9a regulates Sox2 protein stability in breast cancer cells. When G9a lysine methyltransferase activity was chemically inhibited in the ER(+) brea...

  1. Negative in vitro selection identifies the rRNA recognition motif for ErmE methyltransferase

    DEFF Research Database (Denmark)

    Nielsen, A K; Douthwaite, S; Vester, B

    1999-01-01

    the adjacent single-stranded region around A2058. An RNA transcript of 72 nt that displays this motif functions as an efficient substrate for the ErmE methyltransferase. Pools of degenerate RNAs were formed by doping 34-nt positions that extend over and beyond the putative Erm recognition motif within...... contained substitutions at single sites, and these are confined to 12 nucleotide positions. These nucleotides, corresponding to A2051-A2060, C2611, and A2614 in 23S rRNA, presumably comprise the RNA recognition motif for ErmE methyltransferase. The structure formed by these nucleotides is highly conserved...

  2. Radiometric assay for phenylethanolamine N-methyltransferase and catechol O-methyltransferase in a single tissue sample: application to rat hypothalamic nuclei, pineal gland, and heart

    International Nuclear Information System (INIS)

    A simple and highly sensitive method for simultaneous assay of phenylethanolamine N-methyltransferase (PNMT) and catechol O-methyltransferase (COMT) is described. These enzymes are determined in a single tissue homogenate using S-[methyl-3H] adenosyl-L-methionine as methyl donor and sequentially incubating with the substrates phenylethanolamine and epinephrine. The radioactive products of the enzymatic reactions, N-methylphenylethanolamine and metanephrine, are extracted and then separated by thin-layer chromatography. The identity of the reaction products has been established chromatographically and the conditions for both enzymatic reactions in the assay procedure have been defined. Measurement of PNMT activity in the rat pineal gland or in minute fragments of other tissues (e.g., brain nuclei) has not been possible using previously described methods. Activities of PNMT and COMT in the rat pineal gland, various hypothalamic nuclei, and the auricular and ventricular myocardia are herein reported

  3. Broccoli yield and yield quality as affected by gamma rays seeds irradiation and foliar application of some nutrients

    International Nuclear Information System (INIS)

    Two field experiments were carried out during 1999/2000 and 2000/2001 winter growing seasons at the Atomic Energy Authority (AEA) in Inshas, Egypt. The experiment was conducted to study the effect of pre-sowing seeds irradiation with different doses of gamma rays (0, 2, 3 and 4 Gy) and foliar application of different nutrients (Zn, B, S and S+K) on spear diameter, main spear fresh and dry weight per plant, total spear fresh weight per plant, total spear yield besides NPK in leaves at 90 days after transplanting and NPK and total protein content in spears at maturity. In general, exposing broccoli seeds to different gamma ray doses up to 4 Gy prior to sowing increased the above mentioned parameters with different magnitudes comparing with the non-irradiated control plants. There were no significant differences between 3 Gy and 4 Gy treatments during the two growing seasons. With respect to the effect of nutrient application on the studied parameters, all nutrients application significantly increased all the above mentioned parameters. The highest result was detected with B application. Regarding to the interaction of gamma ray with nutrients application, the highest value of all above mentioned parameters was detected with B application and 3 Gy of gamma ray

  4. Broccoli sprout extract prevents diabetic cardiomyopathy via Nrf2 activation in db/db T2DM mice

    Science.gov (United States)

    Xu, Zheng; Wang, Shudong; Ji, Honglei; Zhang, Zhiguo; Chen, Jing; Tan, Yi; Wintergerst, Kupper; Zheng, Yang; Sun, Jian; Cai, Lu

    2016-01-01

    To develop a clinic-relevant protocol for systemic up-regulation of NFE2-related factor 2 (Nrf2) to prevent diabetic cardiomyopathy (DCM), male db/db and age-matched wild-type (WT) mice were given sulforaphane (SFN, an Nrf2 activator) and its natural source, broccoli sprout extract (BSE) by gavage every other day for 3 months, with four groups: vehicle (0.1 ml/10 g), BSE-low dose (estimated SFN availability at 0.5 mg/kg), BSE-high dose (estimated SFN availability at 1.0 mg/kg), and SFN (0.5 mg/kg). Cardiac function and pathological changes (hypertrophy, fibrosis, inflammation and oxidative damage) were assessed by echocardiography and histopathological examination along with Western blot and real-time PCR, respectively. Both BSE and SFN significantly prevented diabetes-induced cardiac dysfunction, hypertrophy and fibrosis. Mechanistically, BSE, like SFN, significantly up-regulated Nrf2 transcriptional activity, evidenced by the increased Nrf2 nuclear accumulation and its downstream gene expression. This resulted in a significant prevention of cardiac oxidative damage and inflammation. For all these preventive effects, BSE at high dose provided a similar effect as did SFN. These results indicated that BSE at high dose prevents DCM in a manner congruent with SFN treatment. Therefore, it suggests that BSE could potentially be used as a natural and safe treatment against DCM via Nrf2 activation. PMID:27457280

  5. Betaine-homocysteine methyltransferase: human liver genotype-phenotype correlation

    Science.gov (United States)

    Feng, Qiping; Kalari, Krishna; Fridley, Brooke L.; Jenkins, Gregory; Ji, Yuan; Abo, Ryan; Hebbring, Scott; Zhang, Jianping; Nye, Monica D.; Leeder, J. Steven; Weinshilboum, Richard. M.

    2010-01-01

    Betaine-homocysteine methyltransferase (BHMT) catalyzes the remethylation of homocysteine. BHMT is highly expressed in the human liver. In the liver, BHMT catalyzes up to 50% of homocysteine metabolism. Understanding the relationship between BHMT genetic polymorphisms and function might increase our understanding of the role of this reaction in homocysteine remethylation and in S-adenosylmethionine-dependent methylation. To help achieve those goals, we measured levels of BHMT enzyme activity and immunoreactive protein in 268 human hepatic surgical biopsy samples from adult subjects as well as 73 fetal hepatic tissue samples obtained at different gestational ages. BHMT protein levels were correlated significantly (p<0.001) with levels of enzyme activity in both fetal and adult tissue, but both were decreased in fetal tissue when compared with levels in the adult hepatic biopsies. To determine possible genotype-phenotype correlations, 12 tag SNPs for BHMT and the closely related BHMT2 gene were selected from SNPs observed during our own gene resequencing studies as well as from HapMap data were used to genotype DNA from the adult hepatic surgical biopsy samples, and genotype-phenotype association analysis was performed. Three SNPs (rs41272270, rs16876512, and rs6875201), located 28 kb upstream, in the 5′-UTR and in intron 1 of BHMT, respectively, were significantly correlated with both BHMT activity (p=3.41E-8, 2.55E-9 and 2.46E-10, respectively) and protein levels (p=5.78E-5, 1.08E-5 and 6.92E-6, respectively). We also imputed 230 additional SNPs across the BHMT and BHMT2 genes, identifying an additional imputed SNP, rs7700790, that was also highly associated with hepatic BHMT enzyme activity and protein. However, none of the 3 genotyped or one imputed SNPs displayed a “shift” during electrophoretic mobility shift assays. These observations may help us to understand individual variation in the regulation of BHMT in the human liver and its possible relationship

  6. Transformation of Broccoli with KTI Gene and the Bioassay for Diamondback Moth Resistance%抗虫相关基因KTI对青花菜的转化及其对小菜蛾抗性的分析

    Institute of Scientific and Technical Information of China (English)

    江汉民; 宋文芹; 刘莉莉; 文正华; 姚星伟; 单晓政; 孙德岭

    2013-01-01

    KTI gene was transformed into broccoli (Brassica oleracea L. ssp. italica) genome via Agrobacterium mediated transformation method using broccoli lines '09LR-11' as explants. Thirteen kanamycin resistant broccoli plants were obtained. PCR analysis with primers specific to KTI gene identified 8 KTI positive plants. Southern blotting analysis with KTI gene as probes showed that the KTI gene was integrated into broccoli genome. RT-PCR analysis indicated that the KTI gene was expressed in transgenic broccoli. Both laboratory and field tests proved that the transgenic broccoli plants were resistant to diamondback moth (Plutella xylostella larvae) .%以青花菜(Brassica oleraceaL.ssp.italica)下胚轴为外植体,通过农杆菌介导的遗传转化方法将来源于杨树的Kunitz型丝氨酸胰蛋白酶抑制剂(KTI)基因导入青花菜品系‘09LR-11’中,获得13株卡那霉素抗性植株.以KTI特异引物对转基因植株进行PCR检测,其中8株为KTI阳性植株.Southernblot分析进一步表明,基因KTI已成功整合到青花菜基因组中.RT-PCR检测表明,KTI在转基因青花菜中已成功表达.通过室内叶片离体试验和田间观察,初步证明转KTI青花菜对小菜蛾幼虫具有一定抗性.

  7. Wolbachia prophage DNA adenine methyltransferase genes in different Drosophila-Wolbachia associations

    DEFF Research Database (Denmark)

    Saridaki, Aggeliki; Sapountzis, Panagiotis; Harris, Harriet L;

    2011-01-01

    . The importance of DNA methylation in cell fate and biology calls for in depth studying of putative methylation-related genes. We present a molecular and phylogenetic analysis of a putative DNA adenine methyltransferase encoded by a prophage in the Wolbachia genome. Two slightly different copies of the gene, met1...

  8. Structural basis for S-adenosylmethionine binding and methyltransferase activity by mitochondrial transcription factor B1.

    Science.gov (United States)

    Guja, Kip E; Venkataraman, Krithika; Yakubovskaya, Elena; Shi, Hui; Mejia, Edison; Hambardjieva, Elena; Karzai, A Wali; Garcia-Diaz, Miguel

    2013-09-01

    Eukaryotic transcription factor B (TFB) proteins are homologous to KsgA/Dim1 ribosomal RNA (rRNA) methyltransferases. The mammalian TFB1, mitochondrial (TFB1M) factor is an essential protein necessary for mitochondrial gene expression. TFB1M mediates an rRNA modification in the small ribosomal subunit and thus plays a role analogous to KsgA/Dim1 proteins. This modification has been linked to mitochondrial dysfunctions leading to maternally inherited deafness, aminoglycoside sensitivity and diabetes. Here, we present the first structural characterization of the mammalian TFB1 factor. We have solved two X-ray crystallographic structures of TFB1M with (2.1 Å) and without (2.0 Å) its cofactor S-adenosyl-L-methionine. These structures reveal that TFB1M shares a conserved methyltransferase core with other KsgA/Dim1 methyltransferases and shed light on the structural basis of S-adenosyl-L-methionine binding and methyltransferase activity. Together with mutagenesis studies, these data suggest a model for substrate binding and provide insight into the mechanism of methyl transfer, clarifying the role of this factor in an essential process for mitochondrial function. PMID:23804760

  9. Suz12 is essential for mouse development and for EZH2 histone methyltransferase activity

    DEFF Research Database (Denmark)

    Pasini, Diego; Bracken, Adrian P; Jensen, Michael R;

    2004-01-01

    SUZ12 is a recently identified Polycomb group (PcG) protein, which together with EZH2 and EED forms different Polycomb repressive complexes (PRC2/3). These complexes contain histone H3 lysine (K) 27/9 and histone H1 K26 methyltransferase activity specified by the EZH2 SET domain. Here we show tha...

  10. Methyltransferase Erm(37) Slips on rRNA to Confer Atypical Resistance in Mycobacterium tuberculosis

    Czech Academy of Sciences Publication Activity Database

    Madsen, Ch. T.; Jakobsen, L.; Buriánková, Karolína; Doucet-Populaire, F.; Perdonet, J. L.; Douthwaite, S.

    2005-01-01

    Roč. 280, č. 47 (2005), s. 38942-38947. ISSN 0021-9258 R&D Projects: GA ČR GA310/03/0292 Institutional research plan: CEZ:AV0Z50200510 Keywords : methyltransferase erm * mycobacterium tuberculosis * rRNA Subject RIV: EE - Microbiology, Virology Impact factor: 5.854, year: 2005

  11. Association of Catechol-O-Methyltransferase (COMT) Polymorphism and Academic Achievement in a Chinese Cohort

    Science.gov (United States)

    Yeh, Ting-Kuang; Chang, Chun-Yen; Hu, Chung-Yi; Yeh, Ting-Chi; Lin, Ming-Yeh

    2009-01-01

    Catechol-O-methyltransferase (COMT) is a methylation enzyme that catalyzes the degradation pathway and inactivation of dopamine. It is accepted widely as being involved in the modulation of dopaminergic physiology and prefrontal cortex (PFC) function. The COMT Val158Met polymorphism is associated with variation in COMT activity. COMT 158Met allele…

  12. Catechol-O-methyltransferase gene methylation and substance use in adolescents: The TRAILS study

    NARCIS (Netherlands)

    L.J. van der Knaap (Lisette); J.M. Schäfer (Johanna); I.H.A. Franken (Ingmar); F.C. Verhulst (Frank); F.V.A. van Oort (Floor); H. Riese (Harriëtte)

    2014-01-01

    textabstractSubstance use often starts in adolescence and poses a major problem for society and individual health. The dopamine system plays a role in substance use, and catechol-O-methyltransferase (COMT) is an important enzyme that degrades dopamine. The Val108/158Met polymorphism modul

  13. Effect of alternative temozolomide schedules on glioblastoma O 6-methylguanine-DNA methyltransferase activity and survival

    OpenAIRE

    Robinson, C G; Palomo, J M; Rahmathulla, G; McGraw, M; Donze, J; L. Liu; Vogelbaum, M A

    2010-01-01

    Background: O 6-methylguanine-DNA methyltransferase (MGMT) expression in glioblastoma correlates with temozolomide resistance. Dose-intense temozolomide schedules deplete MGMT activity in peripheral blood mononuclear cells; however, no published data exist evaluating the effect of temozolomide schedules on intracranial tumour MGMT activity. Methods: Human glioblastoma cells (GBM43) with an unmethylated MGMT promoter were implanted intracranially in immunodeficient rodents. Three weeks later, ...

  14. Clinical Pharmacogenetics Implementation Consortium guidelines for thiopurine methyltransferase genotype and thiopurine dosing

    DEFF Research Database (Denmark)

    Relling, M V; Gardner, E E; Sandborn, W J;

    2011-01-01

    Thiopurine methyltransferase (TPMT) activity exhibits monogenic co-dominant inheritance, with ethnic differences in the frequency of occurrence of variant alleles. With conventional thiopurine doses, homozygous TPMT-deficient patients (~1 in 178 to 1 in 3,736 individuals with two nonfunctional TP...

  15. SABATH Methyltransferases from White Spruce (Picea glauca [Moench] Voss): Gene Cloning, Functional Characterization and Structural Analysis

    Science.gov (United States)

    Known members of the plant SABATH family of methyltransferases have important biological functions by methylating hormones, signaling molecules and other metabolites. While all previously characterized SABATH genes were isolated from angiosperms, in this article, we report on the isolation and funct...

  16. Recognition elements in rRNA for the tylosin resistance methyltransferase RlmA(II)

    DEFF Research Database (Denmark)

    Lebars, Isabelle; Husson, Clotilde; Yoshizawa, Satoko; Douthwaite, Stephen; Fourmy, Dominique

    2007-01-01

    antibiotics. We have previously solved the solution structure of hairpin 35 in the conformation that is recognized by the RlmA(II) methyltransferase from Streptococcus pneumoniae. It was shown that while essential recognition elements are located in hairpin 35, the interactions between RlmA(II) and hairpin 35...

  17. DNA repair methyltransferase (Mgmt) knockout mice are sensitive to the lethal effects of chemotherapeutic alkylating agents.

    NARCIS (Netherlands)

    B.J. Glassner (Brian); G. Weeda (Geert); J.M. Allan (James); J.L.M. Broekhof (Jose'); N.H.E. Carls (Nick); I. Donker (Ingrid); B.P. Engelward (Bevin); R.J. Hampson (Richard); R. Hersmus (Remko); M.J. Hickman (Mark); R.B. Roth (Richard); H.B. Warren (Henry); M.M. Wu (Mavis); J.H.J. Hoeijmakers (Jan); L.D. Samson (Leona)

    1999-01-01

    textabstractWe have generated mice deficient in O6-methylguanine DNA methyltransferase activity encoded by the murine Mgmt gene using homologous recombination to delete the region encoding the Mgmt active site cysteine. Tissues from Mgmt null mice displayed very low O6-methylguanine DNA methyltransf

  18. Functional characterization of cinnamyl alcohol dehydrogenase and caffeic acid O-methyltransferase in Brachypodium distachyon.

    Science.gov (United States)

    Lignin is a significant recalcitrant in the conversion of plant biomass to bioethanol. Cinnamyl alcohol dehydrogenase (CAD) and caffeic acid O-methyltransferase (COMT) catalyze key steps in the pathway of lignin monomer biosynthesis. Brown midrib mutants in Zea mays and Sorghum bicolor with impaired...

  19. Mutations in the DNA methyltransferase gene DNMT3A cause an overgrowth syndrome with intellectual disability

    DEFF Research Database (Denmark)

    Tatton-Brown, Katrina; Seal, Sheila; Ruark, Elise; Harmer, Jenny; Ramsay, Emma; Del Vecchio Duarte, Silvana; Zachariou, Anna; Hanks, Sandra; O'Brien, Eleanor; Aksglaede, Lise; Baralle, Diana; Dabir, Tabib; Gener, Blanca; Goudie, David; Homfray, Tessa; Kumar, Ajith; Pilz, Daniela T; Selicorni, Angelo; Temple, I Karen; Van Maldergem, Lionel; Yachelevich, Naomi; van Montfort, Robert; Rahman, Nazneen

    2014-01-01

    and histone binding. Similar mutations were not present in 1,000 UK population controls (13/152 cases versus 0/1,000 controls; P < 0.0001). Mutation carriers had a distinctive facial appearance, intellectual disability and greater height. DNMT3A encodes a DNA methyltransferase essential for...

  20. Local chromatin microenvironment determines DNMT activity : from DNA methyltransferase to DNA demethylase or DNA dehydroxymethylase

    NARCIS (Netherlands)

    van der Wijst, Monique G. P.; Venkiteswaran, Muralidhar; Chen, Hui; Xu, Guo-Liang; Plosch, Torsten; Rots, Marianne G.

    2015-01-01

    Insights on active DNA demethylation disproved the original assumption that DNA methylation is a stable epigenetic modification. Interestingly, mammalian DNA methyltransferases 3A and 3B (DNMT-3A and -3B) have also been reported to induce active DNA demethylation, in addition to their well-known fun

  1. Clinical Pharmacogenetics Implementation Consortium guidelines for thiopurine methyltransferase genotype and thiopurine dosing

    DEFF Research Database (Denmark)

    Relling, M V; Gardner, E E; Sandborn, W J; Schmiegelow, K; Pui, C-H; Yee, S W; Stein, Paul C.; Carrillo, Maria Berrocal; Evans, W E; Klein, T E

    2011-01-01

    Thiopurine methyltransferase (TPMT) activity exhibits monogenic co-dominant inheritance, with ethnic differences in the frequency of occurrence of variant alleles. With conventional thiopurine doses, homozygous TPMT-deficient patients (~1 in 178 to 1 in 3,736 individuals with two nonfunctional TP...... recommendations (updates at http://www.pharmgkb.org) for azathioprine, mercaptopurine (MP), and thioguanine based on TPMT genotype....

  2. Tyrosine 87 is vital for the activity of human protein arginine methyltransferase 3 (PRMT3)

    Czech Academy of Sciences Publication Activity Database

    Handrková, H.; Petrák, J.; Halada, Petr; Pospíšilová, D.; Čmejla, R.

    2011-01-01

    Roč. 1814, č. 2 (2011), s. 277-282. ISSN 1570-9639 R&D Projects: GA MŠk LC07017 Institutional research plan: CEZ:AV0Z50200510 Keywords : DIAMOND-BLACKFAN ANEMIA * SUBSTRATE-SPECIFICITY * N-METHYLTRANSFERASE Subject RIV: CE - Biochemistry Impact factor: 3.635, year: 2011

  3. A mouse speciation gene encodes a meiotic histone H3 methyltransferase

    Czech Academy of Sciences Publication Activity Database

    Mihola, Ondřej; Trachtulec, Zdeněk; Vlček, Čestmír; Schimenti, J.C.; Forejt, Jiří

    2009-01-01

    Roč. 323, č. 5912 (2009), s. 373-375. ISSN 0036-8075 Institutional research plan: CEZ:AV0Z50520514 Keywords : hybrid sterility * histone H3K4 methyltransferase * Prdm9 Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 29.747, year: 2009

  4. Anti toxic effect of broccoli extract on stannous dichloride toxicity Efeito antitóxico do extrato de brócolis na toxicidade do dicloreto de estanho

    Directory of Open Access Journals (Sweden)

    Betul Cekic

    2012-09-01

    Full Text Available PURPOSE: Since Technetium-99m (99mTc has favorable physical and chemical characteristics, it is widely used radioisotope in Nuclear Medicine. However, stannous dichloride (SnCl2 has been widely used as a reducing agent in labeling procedure of pharmaceutical with radionuclide, it has been realized that SnCl2 have genotoxic and cytotoxic effects on biological systems. In previous studies, it has been shown that some herbal extract can reduce genotoxic and cytotoxic effects of SnCl2. In the present study, it is aimed to evaluate the effect of the broccoli extract on the survival of E. coli ATCC 25922 strain against to toxic effects of SnCl2. METHODS: Broccoli was extracted with methanol extraction. HPLC and TLC analysis of broccoli extract were performed. Then antitoxicity and dose response assays were performed on bacterial strain. RESULTS: The broccoli extract had dose dependent protective effect against SnCl2 toxic effect on E. coli. CONCLUSIONS: The consumption of broccoli may alter the stannous dichloride toxicity. Broccoli extract may use as a new protective strategies against the toxic effect of SnCl2 on patients who were taken 99mTc radiopharmaceuticals.OBJETIVO: Em face de suas características físico-químicas, o Tecnécio-99m (99mTc é um radiofármaco amplamente utilizado na Medicina Nuclear. Todavia, o dicloreto de estanho (SnCl2 tem sido largamente aplicado como um agente redutor no procedimento farmacêutico de marcação com radionuclídeos. Constatou-se que o SnCl2 apresenta efeitos genotóxicos e citotóxicos nos sistemas biológicos. Em estudos prévios, foi demonstrado que alguns extratos de ervas podem reduzir tais efeitos. O estudo atual objetivou avaliar os efeitos do extrato de brócolis na sobrevida da cepa E. coli ATCC 25922, exposta ao efeito tóxico do SnCl2. MÉTODOS: O extrato de brócolis foi obtido mediante extração com metanol. Analises com HPLC e TLC foram efetuadas. Avaliou-se a antitoxicidade e realizou-se um

  5. 辽西地区露地西兰花(绿花椰菜)栽培技术要点%Cultivation Technology of Broccoli in Western Area of Liaoning

    Institute of Scientific and Technical Information of China (English)

    颜津宁; 胡新颖; 杨迎东; 裴新辉; 商旭文; 颜范悦

    2012-01-01

    Planting technology of broccoli in autumn, including variety selection, seedling breeding, planting and managements, prevention and cure for plant diseases and insect pests, harvest were introduced. These techniques would provide references for the production of broccoli in autumn.%该文从品种选择、秧苗培育、整地定植、病虫害防治、收获及贮藏等方面介绍了辽西地区西兰花秋茬栽培技术,以期指导露地秋茬西兰花生产。

  6. 盐碱胁迫对西兰花种子萌发的影响%Effect of Salt and Alkaline Stress on Seed Germination of Broccoli

    Institute of Scientific and Technical Information of China (English)

    徐芬芬

    2012-01-01

    The germination and protection enzyme activities of Brassica oleracea L. var. botrytis L. embryo under the different concentration of NaCl and Na2CO3 stress were examined in this experiment. The results showed that the germination of broccoli was promoted under the low concentration of NaCl (≤20 mmol/L). Germination of broccoli was inhibited under the Na2CO3 stress when concentration of Na+ was the same. SOD and POD activities of embryo under the Na2CO3 stress were lower than the NaCl stress, and MDA content was higher than the NaCl stress. Comprehensive analysis showed that the threshold values of broccoli tolerance to NaCl and Na2CO3 were 445.5 mmol/L and 64.1 mmol/L.%本试验通过采用不同浓度的NaCl和Na2CO3胁迫萌发期的西兰花(Brassica oleracea L.var.botrytis L.)种子,调查了其发芽和胚保护酶活性等指标.研究结果表明,低浓度(≤20 mmol/L)的NaCl胁迫对西兰花各发芽指标有促进作用,在等Na+浓度胁迫下,各Na2CO3胁迫对各发芽指标均表现出抑制作用.Na2CO3胁迫处理胚的SOD和POD活性低于NaCl胁迫处理,MDA含量高于NaCl胁迫处理.综合分析表明,西兰花耐盐阈值和耐碱阈值分别为445.5 mmol/L和64.1 mmol/L.

  7. Physiology and Posttranscriptional Regulation of Methanol:Coenzyme M Methyltransferase Isozymes in Methanosarcina acetivorans C2A ▿ §

    OpenAIRE

    Opulencia, Rina B.; Bose, Arpita; Metcalf, William W.

    2009-01-01

    Methanosarcina species possess three operons (mtaCB1, mtaCB2, and mtaCB3) encoding methanol-specific methyltransferase 1 (MT1) isozymes and two genes (mtaA1 and mtaA2) with the potential to encode a methanol-specific methyltransferase 2 (MT2). Previous genetic studies showed that these genes are differentially regulated and encode enzymes with distinct levels of methyltransferase activity. Here, the effects of promoter strength on growth and on the rate of methane production were examined by ...

  8. Transcriptome and Metabolome Analyses of Glucosinolates in Two Broccoli Cultivars Following Jasmonate Treatment for the Induction of Glucosinolate Defense to Trichoplusia ni (Hübner).

    Science.gov (United States)

    Ku, Kang-Mo; Becker, Talon M; Juvik, John A

    2016-01-01

    Lepidopteran larvae growth is influenced by host plant glucosinolate (GS) concentrations, which are, in turn, influenced by the phytohormone jasmonate (JA). In order to elucidate insect resistance biomarkers to lepidopteran pests, transcriptome and metabolome analyses following JA treatments were conducted with two broccoli cultivars, Green Magic and VI-158, which have differentially induced indole GSs, neoglucobrassicin and glucobrassicin, respectively. To test these two inducible GSs on growth of cabbage looper (Trichoplusia ni), eight neonate cabbage looper larvae were placed onto each of three plants per JA treatments (0, 100, 200, 400 µM) three days after treatment. After five days of feeding, weight of larvae and their survival rate was found to decrease with increasing JA concentrations in both broccoli cultivars. JA-inducible GSs were measured by high performance liquid chromatography. Neoglucobrassicin in Green Magic and glucobrassicin in VI-158 leaves were increased in a dose-dependent manner. One or both of these glucosinolates and/or their hydrolysis products showed significant inverse correlations with larval weight and survival (five days after treatment) while being positively correlated with the number of days to pupation. This implies that these two JA-inducible glucosinolates can influence the growth and survival of cabbage looper larvae. Transcriptome profiling supported the observed changes in glucosinolate and their hydrolysis product concentrations following JA treatments. Several genes related to GS metabolism differentiate the two broccoli cultivars in their pattern of transcriptional response to JA treatments. Indicative of the corresponding change in indole GS concentrations, transcripts of the transcription factor MYB122, core structure biosynthesis genes (CYP79B2, UGT74B1, SUR1, SOT16, SOT17, and SOT18), an indole glucosinolate side chain modification gene (IGMT1), and several glucosinolate hydrolysis genes (TGG1, TGG2, and ESM1) were

  9. 青花菜流通过程中营养品质变化的研究%Study on changes of nutritional quality in circulation of broccoli

    Institute of Scientific and Technical Information of China (English)

    郭李维; 王清; 杨娜; 高丽朴

    2011-01-01

    研究了不同流通温度和包装方式对青花菜外观品质与营养成分的影响。实验结果表明:青花菜在流通过程中随着组织的衰老,其叶绿素、VC、可溶性糖及可溶性蛋白质含量均呈下降趋势。青花菜在5℃中流通比在20℃中能更好地保持其营养成分。利用0.03mm厚的聚乙烯薄膜包装既可避免青花菜失水萎蔫,又可抑制黄化率。20℃无包装处理的青花菜最佳流通时间为1d,20℃PE处理的为2d,5℃无包装处理的为6d,5℃PE处理的为12d。%The effect of different temperature and packaging on the quality and nutrients of broccoli was studied.The results showed that the contents of chlorophyll,ascorbic acid,soluble sugars were decreased with senescence of broccoli in the circulation process.In addition,The broccoli were circulated at 5℃ that could better maintain the quality and nutrients than circulation at 20℃.Packaged with 0.03mm PE could not only prevent the loss of water,but also inhibit the rate of yellowing.Therefore,the broccoli PE packaging could keep a better appearance for 2 days,1 day longer than those directly exposed to the air at 20℃,while cold chain could show great advantage during transportation,the brocoli packed by PE film could store 12d at 5℃,and the unpackaged might keep better quality for 6 days as well.

  10. Standardized cultivation technique for exported pollution-free broccoli%出口西兰花无公害标准化栽培技术

    Institute of Scientific and Technical Information of China (English)

    陈少珍

    2012-01-01

    根据10多年的引种试验、无公害栽培试验和示范推广的结果,总结一套符合出口质量要求、高产优质高效的西兰花无公害标准化栽培技术。%According to the results of introduction,pollution-free cultivation,demonstration and popularization in local area for more than ten years,a standardized pollution-free cultivation technique for broccoli which meet the quality requirements for exportation,with high yield and benefit was summarized in this paper.

  11. Establishment and Optimization of Broccoli Regeneration System%西兰花再生体系的建立与优化

    Institute of Scientific and Technical Information of China (English)

    刘煜; 王红波; 王传琦; 李晶

    2013-01-01

    Objective: Using axenic seedlings as explants, several factors influencing regeneration rate of broccoli were studied and a successful tissue regeneration system was established and optimized. Methods: Using tissue culture mesthod to induce the explant of broccoli yielding bud and root. Results: The study showed that the geno⁃type influenced regeneration rate greatly. The regeneration rate of Zhu-lv and Qing-xiu was above 91% while Ding⁃feng1 is 80.3%; differentiation ability of hypocotyls which had been cultured 6 days was the strongest. When the medium contained 3 mg/L 6-BA and 0.2 mg/L IAA, the differentiation rate of hypocotyls reached the highest lev⁃el and the degree of vitrification was the lowest. In the medium contains 0.2 mg/L IBA or 0.2 mg/L NAA, adventi⁃tious root could be successfully induced from the adventitious buds. Conclusion: Broccoli regeneration system has been successfully established, which laid the foundation for the establishment of broccoli genetic transformation sys⁃tem.%  目的:以西兰花无菌苗为材料,对影响西兰花再生的各种素进行研究,建立并优化西兰花的再生体系.方法:用组织培养法,对西兰花的外植体进行诱导生芽、生根.结果:不同品种的再生率差别较大,珠绿和青秀两个品种的再生率达91%以上,而鼎丰一号仅为80.3%;培养6 d 的下胚轴分化能力最强;当培养基中添加3 mg/L 6-BA 和0.2 mg/L IAA 时,下胚轴的分化率最高,玻璃化程度也降到最低;当培养基中添加0.2 mg/L IBA 或0.2 mg/L NAA时,可成功诱导不定芽生根.结论:建立并优化了西兰花再生体系,为西兰花遗传转化体系的建立奠定了基础.

  12. Research Progress of Broccoli in the Prevention and Treatment of Diseases%西兰花预防和治疗肿瘤研究的新进展

    Institute of Scientific and Technical Information of China (English)

    洪姣; 王佳蕊; 黄春琦; 徐殿琴; 张婷; 许健

    2014-01-01

    In recent years, the morbidity of cancer, cardiovascular disease, diabetes and other diseases were increasing constantly. Meanwhile, people gradually realized that diet played an important role in the prevention or treatment of chronic disease by providing useful elements. Many papers have confirmed that glucosinolates, anthocyanin, vitamins and other beneficial ingredients in broccoli had the function of preventing cancer, diabetes and eye disease, and protecting cardio-vascular and antioxidation. This suggested that increasing our daily diet intake of broccoli could promote the health, prevent diseases. In this paper, we reviewed the medicinal value of broccoli and the influence of cooking and temperature on its useful elements, and illustrated different roles of different effective components of broccoli in the treatment of diseases.%近年来,肿瘤、心血管疾病及糖尿病等疾病的发病率不断上升,人们意识到可以在日常饮食中通过提供有益成分而起到预防或治疗疾病的作用。大量文献证实西兰花中所含有的芥子油苷、花青素、维生素等有益成分具有抗癌、预防糖尿病及眼部疾病、保护心血管抗氧化等作用,提示在日常饮食中增加西兰花的摄入可以促进身体健康,起到防病、治病和改善体质的功效。本文主要综述了西兰花的药用价值以及烹饪、温度等对其有效成分的影响,并总结了西兰花中不同的有效成分治疗疾病的作用。

  13. 打孔包装对西兰花品质的影响%Influence of Perforated Packaging on Broccoli Quality

    Institute of Scientific and Technical Information of China (English)

    杨静; 张蕾

    2012-01-01

    分别在2种厚度的PP薄膜(40,50μm)上以一定间距打不同数量的孔(φ280μm),制得了PP微孔薄膜。将PP微孔薄膜(40μm)制成了自立袋采包装西兰花。通过改变自立袋上微孔的位置,观测袋内气体和西兰花叶绿素含量的变化。结果表明:开孔位置影响袋内O2和CO2气体含量;打孔数量为2个,孔间隔2cm,开孔在西兰花花蕾处时,袋内花蕾处呈低O2和高CO2状态,所包西兰花的叶绿素含量最高,西兰花最新鲜;在环境温度一定的条件下,PP微孔自立袋的厚度、开孔位置、孔径和孔数是影响袋内气体含量的重要因素。%The PP micro-perforated films of 40μm and 50 μm thickness with different number of holes (diameter of 280 μm) in certain distance were prepared. Self-supporting bag was made using the micro-perforated PP film (40μm) for packing broccoli. Change of gas content and chlorophyll content of broccoli was observed by altering micro-perforation's position. The experiment results showed that micro perforation's position affect the content of O2 and CO2 ; when 2 holes separating by 2crn are punched in the broccoli bud, high O2 concentration and CO2 low concentration is produced, content of broccoli's chlorophyll is highest and broccoli is the freshest. The thickness of micro-perforated self-supporting bag, micro-perforation's position, perforation diameter and perforation number are important factors influencing gas content in the package when the environmental temperature is certain.

  14. YccW is the m5C methyltransferase specific for 23S rRNA nucleotide 1962

    DEFF Research Database (Denmark)

    Purta, Elzbieta; O'Connor, Michelle; Bujnicki, Janusz M;

    2008-01-01

    . However, YccW does not methylate assembled 50S subunits, and this is somewhat surprising as the published crystal structures show nucleotide C1962 to be fully accessible at the subunit interface. YccW-directed methylation at nucleotide C1962 is conserved in bacteria, and loss of this methylation in E....... coli marginally reduces its growth rate. YccW had previously eluded identification because it displays only limited sequence similarity to the m(5)C methyltransferases RsmB and RsmF and is in fact more similar to known m(5)U (5-methyluridine) RNA methyltransferases. In keeping with the previously...... proposed nomenclature system for bacterial rRNA methyltransferases, yccW is now designated as the rRNA large subunit methyltransferase gene rlmI....

  15. N-6-Adenine-Specific DNA Methyltransferase 1 (N6AMT1) Polymorphisms and Arsenic Methylation in Andean Women

    OpenAIRE

    Harari, Florencia; Engström, Karin; Concha, Gabriela; Colque, Graciela; Vahter, Marie; Broberg, Karin

    2013-01-01

    BACKGROUND: In humans, inorganic arsenic is metabolized to methylated metabolites mainly by arsenic (+3 oxidation state) methyltransferase (AS3MT). AS3MT polymorphisms are associated with arsenic metabolism efficiency. Recently, a putative N-6-adenine-specific DNA methyltransferase 1 (N6AMT1) was found to methylate arsenic in vitro. OBJECTIVE: We evaluated the role of N6AMT1 polymorphisms in arsenic methylation efficiency in humans. METHODS: We assessed arsenic methylation efficiency in 188 w...

  16. MOLECULAR AND FUNCTIONAL CHARACTERIZATION OF A JUVENILE HORMONE ACID METHYLTRANSFERASE EXPRESSED IN THE CORPORA ALLATA OF MOSQUITOES

    OpenAIRE

    Mayoral, Jaime G.; Nouzova, Marcela; Yoshiyama, Michiyo; Shinoda, Tetsuro; Hernandez-Martinez, Salvador; Dolghih, Elena; Turjanski, Adrian G; Roitberg, Adrian E.; Priestap, Horacio; Perez, Mario; Mackenzie, Lucy; Li, Yiping; Noriega, Fernando G.

    2008-01-01

    A juvenile hormone acid methyltransferase (JHAMT) was isolated as an abundant EST in a library of the corpora allata of the adult female mosquito Aedes aegypti. Its full-length cDNA encodes a 278-aa protein that has 43 % amino acid identity with BmJHAMT, a juvenile hormone acid methyltransferase previously cloned from Bombyx mori. Heterologous expression produced a recombinant protein that metabolizes farnesoic acid (FA) into methyl farnesoate, as well as juvenile hormone acid into juvenile h...

  17. The Histone Methyltransferase Inhibitor A-366 Uncovers a Role for G9a/GLP in the Epigenetics of Leukemia

    OpenAIRE

    Pappano, William N.; Guo, Jun; He, Yupeng; Ferguson, Debra; Jagadeeswaran, Sujatha; Osterling, Donald J.; Gao, Wenqing; Spence, Julie K.; Pliushchev, Marina; Sweis, Ramzi F.; Buchanan, Fritz G.; Michaelides, Michael R.; Shoemaker, Alexander R.; Tse, Chris; Chiang, Gary G.

    2015-01-01

    Histone methyltransferases are epigenetic regulators that modify key lysine and arginine residues on histones and are believed to play an important role in cancer development and maintenance. These epigenetic modifications are potentially reversible and as a result this class of enzymes has drawn great interest as potential therapeutic targets of small molecule inhibitors. Previous studies have suggested that the histone lysine methyltransferase G9a (EHMT2) is required to perpetuate malignant...

  18. Inhibition of G9a Histone Methyltransferase Converts Bone Marrow Mesenchymal Stem Cells to Cardiac Competent Progenitors

    OpenAIRE

    Jinpu Yang; Keerat Kaur; Li Lin Ong; Eisenberg, Carol A; Leonard M. Eisenberg

    2015-01-01

    The G9a histone methyltransferase inhibitor BIX01294 was examined for its ability to expand the cardiac capacity of bone marrow cells. Inhibition of G9a histone methyltransferase by gene specific knockdown or BIX01294 treatment was sufficient to induce expression of precardiac markers Mesp1 and brachyury in bone marrow cells. BIX01294 treatment also allowed bone marrow mesenchymal stem cells (MSCs) to express the cardiac transcription factors Nkx2.5, GATA4, and myocardin when subsequently exp...

  19. Antiamnesic Effect of Broccoli (Brassica oleracea var. italica) Leaves on Amyloid Beta (Aβ)1-42-Induced Learning and Memory Impairment.

    Science.gov (United States)

    Park, Seon Kyeong; Ha, Jeong Su; Kim, Jong Min; Kang, Jin Yong; Lee, Du Sang; Guo, Tian Jiao; Lee, Uk; Kim, Dae-Ok; Heo, Ho Jin

    2016-05-01

    To examine the antiamnesic effects of broccoli (Brassica oleracea var. italica) leaves, we performed in vitro and in vivo tests on amyloid beta (Aβ)-induced neurotoxicity. The chloroform fraction from broccoli leaves (CBL) showed a remarkable neuronal cell-protective effect and an inhibition against acetylcholinesterase (AChE). The ameliorating effect of CBL on Aβ1-42-induced learning and memory impairment was evaluated by Y-maze, passive avoidance, and Morris water maze tests. The results indicated improving cognitive function in the CBL group. After the behavioral tests, antioxidant effects were detected by superoxide dismutase (SOD), oxidized glutathione (GSH)/total GSH, and malondialdehyde (MDA) assays, and inhibition against AChE was also presented in the brain. Finally, oxo-dihydroxy-octadecenoic acid (oxo-DHODE) and trihydroxy-octadecenoic acid (THODE) as main compounds were identified by quadrupole time-of-flight ultraperformance liquid chromatography (Q-TOF UPLC-MS) analysis. Therefore, our studies suggest that CBL could be used as a natural resource for ameliorating Aβ1-42-induced learning and memory impairment. PMID:27079470

  20. Characterization and Identification of Gamma-Irradiated Kimchi Cabbage and Broccoli by Electron Spin Resonance Spectroscopy using Different Sample Pre-treatments

    International Nuclear Information System (INIS)

    Electron spin resonance (ESR) spectroscopy of gamma-irradiated fresh broccoli and kimchi cabbage was conducted to identify their irradiation history. Different pretreatments, such as freeze-drying (FD), oven-drying (OD), alcoholic-drying (ALD), and water-washing and alcoholic-drying (WAD) were used to lower the moisture contents of the samples prior to ESR analysis. The non-irradiated samples exhibited a single central signal (g0 = 2.0007) with clear effect of Mn2+, especially in kimchi cabbage. Upon irradiation, there was an increase in the intensity of the central signal, and two side peaks, mutually spaced at 6 mT, were also observed. These side peaks with g1 (left) = 2.023 and g2 (right) = 1.985 were attributed to radiation-induced cellulose radicals. Leaf and stem in broccoli, and root and stem in kimchi cabbage provided good ESR signal responses upon irradiation. The signal noise was reduced in case of ALD and WAD pretreatments, particularly due to Mn2+ signals. The ALD treatment was found most feasible to detect the improved ESR spectra in the irradiated samples. (author)

  1. Laser synthesized super-hydrophobic conducting carbon with broccoli-type morphology as a counter-electrode for dye sensitized solar cells

    Science.gov (United States)

    Gokhale, Rohan; Agarkar, Shruti; Debgupta, Joyashish; Shinde, Deodatta; Lefez, Benoit; Banerjee, Abhik; Jog, Jyoti; More, Mahendra; Hannoyer, Beatrice; Ogale, Satishchandra

    2012-10-01

    A laser photochemical process is introduced to realize superhydrophobic conducting carbon coatings with broccoli-type hierarchical morphology for use as a metal-free counter electrode in a dye sensitized solar cell. The process involves pulsed excimer laser irradiation of a thin layer of liquid haloaromatic organic solvent o-dichlorobenzene (DCB). The coating reflects a carbon nanoparticle-self assembled and process-controlled morphology that yields solar to electric power conversion efficiency of 5.1% as opposed to 6.2% obtained with the conventional Pt-based electrode.A laser photochemical process is introduced to realize superhydrophobic conducting carbon coatings with broccoli-type hierarchical morphology for use as a metal-free counter electrode in a dye sensitized solar cell. The process involves pulsed excimer laser irradiation of a thin layer of liquid haloaromatic organic solvent o-dichlorobenzene (DCB). The coating reflects a carbon nanoparticle-self assembled and process-controlled morphology that yields solar to electric power conversion efficiency of 5.1% as opposed to 6.2% obtained with the conventional Pt-based electrode. Electronic supplementary information (ESI) available: Materials and equipment details, solar cell fabrication protocol, electrolyte spreading time measurement details, XPS spectra, electronic study, film adhesion test detailed analysis and field emission results. See DOI: 10.1039/c2nr32082g

  2. Effectiveness of a spontaneous carvacrol nanoemulsion against Salmonella enterica Enteritidis and Escherichia coli O157:H7 on contaminated broccoli and radish seeds.

    Science.gov (United States)

    Landry, Kyle S; Micheli, Sean; McClements, David Julian; McLandsborough, Lynne

    2015-10-01

    The incidence of foodborne illness associated with the consumption of fresh produce has continued to increase over the past decade. Sprouts, such as mung bean, alfalfa, radish, and broccoli, are minimally processed and have been sources for foodborne illness. Currently, a 20,000 ppm calcium hypochlorite soak is recommended for the treatment of sprouting seeds. In this study, the efficacy of an antimicrobial carvacrol nanoemulsion was tested against Salmonella enterica subspecies enterica serovar Enteritidis (ATCC BAA-1045) or EGFP expressing Escherichia coli O157:H7 (ATCC 42895) contaminated sprouting seeds. Antimicrobial treatments were performed by soaking inoculated seeds in nanoemulsions (4000 or 8000 ppm) for 30 or 60 min. Following treatment, surviving cells were determined by performing plate counts and/or Most Probable Number (MPN) enumeration. Treated seeds were sprouted and tested for the presence of pathogens. Treatment successfully inactivated low levels (2 and 3 log CFU/g) of S. Enteritidis and E. coli on radish seeds when soaked for 60 min at concentrations ≥4000 (0.4%) ppm carvacrol. This treatment method was not affective on contaminated broccoli seeds. Total sprout yield was not influenced by any treatments. These results show that carvacrol nanoemulsions may be an alternative treatment method for contaminated radish seeds. PMID:26187822

  3. Study on Ultrasonic Extraction Technique of Broccoli Polysaccharide%超声波法提取西兰花多糖的工艺研究

    Institute of Scientific and Technical Information of China (English)

    郎艳婷; 刘波; 张振宇; 郝明月; 苏万平

    2012-01-01

    采用超声波辅助法提取西兰花多糖,通过单因素试验与正交试验对提取工艺进行优化,以期确定西兰花多糖的最佳提取工艺.结果表明,超声波辅助法提取西兰花多糖的最佳工艺参数为:超声时间50min,超声波功率80W,提取温度60℃,料液比1∶50(mL/g),多糖的得率为8.24%.该试验方法简单、易行,可为西兰花多糖的工业化生产提供参考.%To determine the best broccoli polysaccharide extraction process with the ultrasonic assisted-extraction technology. The technique was optimized by a single factor experiments and orthogonal test. The results showed that the optimum extraction parameters of polysaccharides was that ultrasonic time for50 min, 80 W ultrasonic power at 60℃, ratio of liquid feed 1:50 (ml/g), and polysaccharide yield was 8.24%. The test method is simple, easy, and can provide a reference for the industrial production of broccoli polysaccharide.

  4. Study of Cleome Gynandra Extract on Broccoli Preservation%白花菜提取物对花椰菜保鲜效果的研究

    Institute of Scientific and Technical Information of China (English)

    曾维丽; 张臻; 刘少阳; 王飞

    2015-01-01

    论文研究了白花菜提取物对花椰菜中微生物、褐变指数和VC含量的影响。结果表明,白花菜提取物能有效抑制微生物的生长,有效减缓花椰菜的褐变速度和VC的降解速度,较好地保持花椰菜的品质,其中1.0 g/L白花菜提取物的保鲜效果最好。%The effects of Cleome gynandra extract on total number of colony, as well as browning index and vitamin C content of Broccoli were studied. The results showed that Cleome gynandra extract could effectively eliminate the microorganism, delay the browning rate and slow down the degradation of vitamin C. 1.0g/L Cleome gynandra extract was shown to be the best to fresh-keeping and storage quality of broccoli.

  5. Broccoli-derived phytochemicals indole-3-carbinol and 3,3’-diindolylmethane exert concentration-dependent pleiotropic effects on prostate cancer cells: Comparison with other cancer preventive phytochemicals

    Science.gov (United States)

    In the present studies, we utilized prostate cancer cell culture models to elucidate the mechanisms of action of broccoli-derived phytochemicals 3, 3’-diindolylmethane (DIM) and indole-3-carbinol (I3C). We found DIM and I3C at 1-5 uM inhibited androgen and estrogen-mediated pathways and induced a x...

  6. 花椰菜在不同贮藏温度下的品质变化及货架期预测%The Quality Changes of Broccoli Stored at Different Temperatures and Its Shelf Life Prediction

    Institute of Scientific and Technical Information of China (English)

    宋晨; 刘宝林; 王欣; 董庆利; 孙雅琼

    2009-01-01

    Broccoli is nutritious and tasty.In this study,Broccoli was stored at 0℃,6℃,24℃ respectively.The sensory quality,water loss and ascorbic acid content were measured at various time intervals to study the effects of temperature on the quality of broccoli.It was found that the degradation of ascorbic acid content followed the first order reaction kinetics.The activation energy of ascorbic acid was determined.Taking 50% ascorbic acid loss as the threshold,a mathematic model for predicting the shelf life of broccoli was established.%花椰菜是一种清香爽脆、营养丰富的蔬菜,试验在0、6℃及室温24℃下贮藏花椰菜,从感官品质、失水率及V_C含量3方面研究温度对其品质的影响.确定了花椰菜中V_C含量下降遵循一级动力学反应,并据此计算出反应的表观活化能Ea进一步推算出以V_C含量下降50%为货架期终点时,花椰菜的货架期计算公式.

  7. 不同贮藏性青花菜花球的抗氧化特性%Antioxidative Properties of Broccoli Florets of Cultivars with Different Storability

    Institute of Scientific and Technical Information of China (English)

    李进才; 赵习姮

    2011-01-01

    为研究青花菜(Brassica oleracea L.Var.italica)花球的抗氧化特性与贮藏性的关系,选取贮藏性不同的春季2个品种(“绿颜”和“城堡”)和秋季2个品种(“爱国者”和“高地”),将采后花球贮藏于20℃条件下,在贮藏开始时和4d后,测定了花球呼吸速率、衰老生理指标(叶绿素、蛋白质和丙二醛(MDA)含量)和抗氧化特性指标(活性氧超氧阴离子和过氧化氢含量,抗氧化酶超氧化物岐化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)和抗坏血酸过氧化氢酶(APX)活性,抗氧化物质抗坏血酸(AsA)和β-胡萝卜素(β-Car)含量).“绿颜”和“爱国者”贮藏开始时,呼吸速率低,活性氧含量少,抗氧化酶活性低;贮藏4d后,叶绿素和蛋白质含量高,MDA含量少,SOD、CAT和POD活性上升幅度大,APX活性和AsA、β-Car含量下降幅度小.结果表明:“绿颜”比“城堡”、“爱国者”比“高地”衰老慢,耐贮藏;耐贮藏青花菜品种的花球具有采收时抗氧化酶活性低,贮藏期间SOD、CAT和POD活性上升幅度大,APX活性和抗氧化物质含量下降幅度小的抗氧化特性.%In this work, relationship between the storability of broccoli {Brassica oleracea L. Var. Italica) florets and their antioxidative properties was investigated. Broccoli florets of four cultivars from the spring "Green face" and "Castle" and autumn "Patriot" and "Heights" with different storage properties were harvested and stored at 20 ℃ until use. Respiratory rate of broccoli florets and senescence physiological indicators (chlorophyll, protein and malondialdehyde (MDA) contents) and antioxidative properties indicators (reactive oxygen species contents of super-oxide and hydrogen peroxide, antioxidative enzymes activities of superoxide dismutase(SOD), catalase(CAT), peroxidase (POD), ascorbate peroxidase (APX), anti-oxidative substances contents of ascorbic acid (AsA) and β-carotene (β-Car)) of broccoli flower buds

  8. Studies on the Preparation and Antioxidant Activity of Lactobacillus-fermented Broccoli Beverage%西兰花乳酸菌饮料的制备及其抗氧化作用研究

    Institute of Scientific and Technical Information of China (English)

    董青青; 胡海杰; 肖文佳; 安兴娟; 姬阿美; 杜稳; 张同存; 罗学刚

    2015-01-01

    Using broccoli as main material,green-protecting condition,formulations,stability,fermentative strain and fermentation process of broccoli lactobacillus beverage were researched in this paper. The optimized technology was ultimately set up as after broccoli was stripped and sliced,Using 50% broccoli juice,10% orange juice,5% flavedo,6% sucrose,0.2% citric acid,0.25% salt and 0.2% CMC-Na as materials fermenting with lactobacillus plantarum for 4 hours at 30℃in anaerobic box. Broccoli lactobacillus beverage made by this technology has an unique flavor and nice trait. Additionally,broccoli has antioxidant activity,allows people to avoid free radical damage,reduce the incidence of cancer. the efficiency of antioxidant and antiradical of the Pre-fermentation and fermented broccoli was determined by the DPPH.%以西兰花为主要原料,通过对西兰花乳酸菌发酵饮料的原料护色方法、配方、稳定性、发酵菌种及发酵过程进行分析,最终优化建立了西兰花乳酸菌发酵制备工艺为:西兰花汁50%,橙汁10%,橙皮5%,蔗糖6%,柠檬酸0.2%,盐0.25%,CMC-Na为0.2%作为发酵原料,按照3%量接种植物乳杆菌后,在厌氧培养箱中30℃发酵4h。以此工艺生产的西兰花乳酸菌发酵饮料具有独特的风味和良好的性状。另外,西兰花具有抗氧化性,可以使人们避免受到自由基的伤害,降低发病几率。利用DPPH法对发酵前后西兰花的抗氧化、清除自由基能力进行了测定。西兰花汁发酵前对DPPH的清除率为67.09%,发酵后对DPPH的清除率为82.85%,结果表明发酵后比发酵前西兰花抗氧化能力明显增强。

  9. LED复合光处理对西兰花低温保鲜效果的影响%Effect of LED Composite Light on Preservation of Broccoli during Cold Storage

    Institute of Scientific and Technical Information of China (English)

    李宁; 阎瑞香; 张娜

    2015-01-01

    In order to study on effect of different treat of LED composite light on preservation of broccoli ,fresh broccoli was used as the main materials .By measuring the indicators at 4 ℃ low temperature conditions , such as sensory quality ,vitamin C content ,chlorophyll content and broccoli ethylene production rate ,effect of LED compos-ite light on preservation for broccoli was determined .The results showed that treatment with LED composite of red and blue was better for the quality of broccoli compared with non-light treatment.The former could extend storage time by 10-15 d.Meanwhile,it could got a higher score of sensory evaluation ,prevented the rapid decline in im-portant nutrient substance vitamin C content of broccoli ,delayed the time of the evolution of ethylene and the ap-pearance of respiration peak , decreased the respiration variable peak , and reduced membrane lipid peroxidation damage of broccoli during storage .%为研究不同LED复合光处理对西兰花保鲜效果的影响。以新鲜西兰花为材料,通过测定4℃低温条件下,西兰花的感官品质、维生素C含量、叶绿素含量、乙烯生成速率等各项指标,研究LED红蓝、LED红绿复合光处理对西兰花保鲜效果的影响。结果表明,与无光处理方式相比, LED红蓝复合光处理效果显著,不仅延长了西兰花保鲜期10~15 d左右,而且较好地保持了西兰花原有的外观品质,阻止了西兰花贮藏期间重要营养物质Vc含量的快速流失,延缓了贮藏期间乙烯释放量峰值和呼吸跃变出现的时间,显著降低了呼吸跃变的峰值,减少了膜脂过氧化对西兰花造成的损伤。

  10. Evaluation on the Apparent Metabolic Energy and Apparent Heal Amino Acid Digestibility of Broccoli LPC and Broccoli Residues for Broilers%西兰花叶蛋白和茎叶粉对肉仔鸡代谢能和回肠氨基酸表观消化率的影响

    Institute of Scientific and Technical Information of China (English)

    杨桂芹; 梁丽萍; 李建涛; 刘国华; 张姝; 郑爱娟

    2011-01-01

    通过2个试验测定了西兰花叶蛋白和茎叶粉在肉仔鸡中的氮校正表观代谢能和回肠氨基酸的表观消化率,为合理利用西兰花资源提供科学依据和数据参考.试验1选用体重相近的22日龄健康肉仔鸡144只,随机分成3个组,每组8个重复,公母各50%.其中1组饲喂基础日粮,另2组分别饲喂试验日粮(西兰花叶蛋白30%替代基础日粮,西兰花茎叶粉35%替代基础日粮).选用TiO2为外源指示剂进行代谢试验;试验2选用体重相近的26日龄健康肉仔鸡96只,随机分为2组,每组8个重复,公母各50%.选用TiO2为外源指示剂,采用回肠末端法进行代谢试验.结果表明:西兰花叶蛋白和茎叶粉在肉仔鸡中的表观代谢能分别为8.86MJ·kg-1和2.48 MJ·kg-1,氮校正表观代谢能分别为8.14 MJ·kg-1和1.86 MJ·kg-1;西兰花叶蛋白和茎叶粉在肉仔鸡中4种限制性氨基酸的回肠末端表观消化率分别为赖氨酸(48.11±4.28)%和(73.00±6.64)%,蛋氨酸(39.51±4.60)%和(79.18±3.91)%,苏氨酸(33.11±4.66)%和(59.38±6.25)%,胱氨酸(33.47±2.94)%和(65.82±4.46)%;存肉仔鸡中17种氨基酸的回肠末端表观消化率的平均值为(39.57±4.45)%和(71.75±4.85)%.可见,西兰花茎叶粉在肉仔鸡中的表观代谢能较低,西兰花叶蛋白在肉仔鸡中回肠末端氨基酸的消化率较低.%To assess the apparent metabolic energy and apparent ileal amino acid digestibility of broccoli leaf protein concentrate (LPC) and broccoli residues for broilers, two experiments were carried out. In experiment one,one hundred and forty-four 22-day-old broilers were allocated randomly into three treatments,each of them contained eight repetitions, male and female half and haft. Titanium dioxide was used as the exogenous indicator to carry out the metabolic test, One group was fed basic diet, other two groups were fed test diets in which the basic diet was replaced by 30% broccoli LPC and 35% broccoli residues respectively. In

  11. Discovery of sphingosine 1-O-methyltransferase in rat kidney and liver homogenates

    Institute of Scientific and Technical Information of China (English)

    Santosh J SACKET; Dong-soon IM

    2008-01-01

    Aim:To characterize sphingosine methyltransferase in rat tissues.Methods:By using S-adenosyl-L-(methyl-3H) methionine,enzymatic activity was measured in the rat liver and kidney homogenates.Results:The optimum pH and reaction time for the enzyme assay were pH 7.8 and 1 h.ZnCl2 inhibited the activity,but not MgCl2,CaCl2,CoCl2,or NiCl2.In the kidney homogenate,enzymatic activity was detectable in the cytosol and all membrane fractions from the plasma membrane and other organelles; however,in the liver homogenate,enzymatic activity was detectable in all membrane fractions,but not in the cytosol.We also tested the enzymatic activity with structurally-modified sphingosine derivatives.Conclusion:We found sphingosine l-O-methyltransferase activity in the rat liver and kidney homogenates.

  12. Super-resolution optical DNA Mapping via DNA methyltransferase-directed click chemistry

    DEFF Research Database (Denmark)

    Vranken, Charlotte; Deen, Jochem; Dirix, Lieve;

    2014-01-01

    We demonstrate an approach to optical DNA mapping, which enables near single-molecule characterization of whole bacteriophage genomes. Our approach uses a DNA methyltransferase enzyme to target labelling to specific sites and copper-catalysed azide-alkyne cycloaddition to couple a fluorophore...... to the DNA. We achieve a labelling efficiency of ∼70% with an average labelling density approaching one site every 500 bp. Such labelling density bridges the gap between the output of a typical DNA sequencing experiment and the long-range information derived from traditional optical DNA mapping. We lay...... the foundations for a wider-scale adoption of DNA mapping by screening 11 methyltransferases for their ability to direct sequence-specific DNA transalkylation; the first step of the DNA labelling process and by optimizing reaction conditions for fluorophore coupling via a click reaction. Three of 11 enzymes...

  13. Crystallization and preliminary X-ray diffraction studies of a catechol-O-methyltransferase/inhibitor complex

    International Nuclear Information System (INIS)

    Catechol-O-methyltransferase has been co-crystallized with a novel inhibitor, which has potential therapeutic application in the Parkinson’s disease therapy. Inhibitors of the enzyme catechol-O-methyltransferase (COMT) are used as co-adjuvants in the therapy of Parkinson’s disease. A recombinant form of the soluble cytosolic COMT from rat has been co-crystallized with a new potent inhibitor, BIA 8-176 [(3,4-dihydroxy-2-nitrophenyl)phenylmethanone], by the vapour-diffusion method using PEG 6K as precipitant. Crystals diffract to 1.6 Å resolution on a synchrotron-radiation source and belong to the monoclinic space group P21, with unit-cell parameters a = 52.77, b = 79.63, c = 61.54 Å, β = 91.14°

  14. Crystallization and preliminary X-ray diffraction studies of a catechol-O-methyltransferase/inhibitor complex

    Energy Technology Data Exchange (ETDEWEB)

    Rodrigues, M. L. [Instituto de Tecnologia Química e Biológica (ITQB), Universidade Nova de Lisboa, Av. República, Apt. 127, 2781-901 Oeiras (Portugal); Bonifácio, M. J.; Soares-da-Silva, P. [Department of Research and Development, BIAL, 4785 S. Mamede do Coronado (Portugal); Carrondo, M. A.; Archer, M., E-mail: archer@itqb.unl.pt [Instituto de Tecnologia Química e Biológica (ITQB), Universidade Nova de Lisboa, Av. República, Apt. 127, 2781-901 Oeiras (Portugal)

    2005-01-01

    Catechol-O-methyltransferase has been co-crystallized with a novel inhibitor, which has potential therapeutic application in the Parkinson’s disease therapy. Inhibitors of the enzyme catechol-O-methyltransferase (COMT) are used as co-adjuvants in the therapy of Parkinson’s disease. A recombinant form of the soluble cytosolic COMT from rat has been co-crystallized with a new potent inhibitor, BIA 8-176 [(3,4-dihydroxy-2-nitrophenyl)phenylmethanone], by the vapour-diffusion method using PEG 6K as precipitant. Crystals diffract to 1.6 Å resolution on a synchrotron-radiation source and belong to the monoclinic space group P2{sub 1}, with unit-cell parameters a = 52.77, b = 79.63, c = 61.54 Å, β = 91.14°.

  15. Targeting histone methyltransferases and demethylases in clinical trials for cancer therapy.

    Science.gov (United States)

    Morera, Ludovica; Lübbert, Michael; Jung, Manfred

    2016-01-01

    The term epigenetics is defined as heritable changes in gene expression that are not due to alterations of the DNA sequence. In the last years, it has become more and more evident that dysregulated epigenetic regulatory processes have a central role in cancer onset and progression. In contrast to DNA mutations, epigenetic modifications are reversible and, hence, suitable for pharmacological interventions. Reversible histone methylation is an important process within epigenetic regulation, and the investigation of its role in cancer has led to the identification of lysine methyltransferases and demethylases as promising targets for new anticancer drugs. In this review, we describe those enzymes and their inhibitors that have already reached the first stages of clinical trials in cancer therapy, namely the histone methyltransferases DOT1L and EZH2 as well as the demethylase LSD1. PMID:27222667

  16. Aflatoxin B1 induced upregulation of protein arginine methyltransferase 5 in human cell lines.

    Science.gov (United States)

    Ghufran, Md Sajid; Ghosh, Krishna; Kanade, Santosh R

    2016-09-01

    The exposure of naturally occurring mycotoxins affects human health and play a vital role in cancer initiation and progression. Aflatoxin B1 is a difuranocoumarin mycotoxin, classified as a group I carcinogen. The present study was conducted to assess the effect of aflatoxin B1 on epigenetic regulatory proteins. The protein arginine methyltransferase 5 expression was induced upon aflatoxin B1 treatment in a dose and time dependent manner. Further global arginine methylation was also increased in the same manner. This is the first report showing the induction of epigenetic regulatory protein, protein arginine methyltransferase 5 upon aflatoxin B1 treatment. Further study is required to establish the detailed pathway of PRMT5 induction. PMID:27242039

  17. 外源硝普钠处理对青花菜贮藏过程中品质的影响%The Effect of Exogenous Sodium Nitroprusside Treatment on Broccoli Quality during the Storage

    Institute of Scientific and Technical Information of China (English)

    史君彦; 王清; 高丽朴; 杨娜

    2012-01-01

    [Objective] The aim was to provide certain theoretical basis for the protection of broccoli. [ Method] Exogenous sodium nitroprusside was used to in the experiment. Under the condition of 20 ℃ and RH 95% , the optimal SNP concentration of broccoli was determined. [ Result ] The exogenous SNP could effectively maintain the quality of broccoli, inhabited the yellowing of broccoli and the increasing of cell membrane permeability, and maintain the soluble solids content, decreased the degradation rate of chlorophyll and delayed the loss of Vc. [ Conclusion] Exogenous sodium nitroprusside can effectively maintain the quality of broccoli to keep it fresh.%[目的]为常温下青花菜的科学保鲜提供理论依据.[方法]试验采用外源硝普钠(SNP)作为NO供体处理青花菜,并在常温(20℃)、RH95%条件下贮藏观察,确定最优SNP浓度下的青花菜相应品质指标.[结果]200μmol/L外源SNP处理可有效地延缓青花菜的黄化,抑制细胞膜透性的增加,维持了其可溶性固形物含量,降低了叶绿素的降解速率,延缓了Vc的流失,有效保持了青花菜的品质.[结论]适宜浓度的外源硝普钠处理能够有效保持青花菜的品质,有较好的保鲜效果.

  18. 冰温结合气调保鲜膜对西兰花保鲜效果的影响%The fresh-keeping effect of ice-temperature combined with plastic film on broccoli

    Institute of Scientific and Technical Information of China (English)

    张娜; 关文强; 张怡

    2012-01-01

    研究冰温条件下不同厚度保鲜膜包装的西兰花品质变化情况,为西兰花的长期保鲜提供依据。结果表明,冰温库的温度控制性能良好,在(-0.1±0.2)℃之间波动;保鲜膜结合冰温贮藏对西兰花具有保鲜效果,保鲜膜越厚,保鲜效果越明显,能够有效延长西兰花贮藏时间,可达110d,比普通冷库有效延长贮藏期30d,同时能够防止西兰花霉烂,保持Vc和可溶性固形物含量,抑制色素降解。%Under the condition of ice-temperature, the quality of broccoli packaged by different thickness of plastic film was test, which would provide the basis for long-term preservation of broccoli. The results showed that the ice-temperature storage had a stable temperature at -0.1 ℃ and the fluctuations is 0.2℃ . The ice-temperature storage combined with plastic-film package played a significant role in preservation of broccoli, the plastic film is thicker, the effect of preservation is more obvious, which can effectively extend the storage time of broccoli, up to 110 d, that is 30 d longer than the normal cold storage, meanwhile prevented broccoli to rot, maintained the Vc and soluble solids content, inhibited anthocyanin degraded.

  19. Effects of Different Pollination Periods on Seed Yield of Broccoli Self-incompatible Lines%不同授粉时期对青花菜自交不亲和系种子产量的影响

    Institute of Scientific and Technical Information of China (English)

    李易蓉; 木万福; 袁建民; 但忠; 苏银玲; 杨龙

    2013-01-01

    为了提高青花菜自交不亲和系种子产量,研究了授粉时期对不同熟性青花菜自交不亲和系结荚率、平均单荚籽粒数、种子千粒质量和种子产量的影响。研究结果表明,不同授粉时期对青花菜自交不亲和系种子生产有显著差异,早熟青花菜自交不亲和系在末花期授粉的结荚率、单荚籽粒数、种子千粒质量、种子产量最高,而中熟、晚熟青花菜自交不亲和系在盛花期授粉结荚率、单荚籽粒数、种子千粒质量、种子产量最高。%In order to increase the seed yield of self-incompatible lines of broccoli, we studied the effects of different pollination periods on podding rate, grain number of a single pod, thousand seed weight and seed yield of the self -incompatible lines of broccoli in this paper. The results showed that, different pollination periods affected the seed production of the self-incompatible lines of broccoli significantly. The late flowering period was the best pollination period for early-maturing broccoli cultivar, while the full-bloom period was best for mid-maturing and late-maturing cultivars, by doing so, we could obtain the highest podding rate, grain number of a single pod, thousand seed weight and seed yield of the three different kinds of broccoli cultivars.

  20. Structural basis for S-adenosylmethionine binding and methyltransferase activity by mitochondrial transcription factor B1

    OpenAIRE

    Guja, Kip E.; Venkataraman, Krithika; Yakubovskaya, Elena; Hui SHI; Mejia, Edison; Hambardjieva, Elena; Karzai, A. Wali; Garcia-Diaz, Miguel

    2013-01-01

    Eukaryotic transcription factor B (TFB) proteins are homologous to KsgA/Dim1 ribosomal RNA (rRNA) methyltransferases. The mammalian TFB1, mitochondrial (TFB1M) factor is an essential protein necessary for mitochondrial gene expression. TFB1M mediates an rRNA modification in the small ribosomal subunit and thus plays a role analogous to KsgA/Dim1 proteins. This modification has been linked to mitochondrial dysfunctions leading to maternally inherited deafness, aminoglycoside sensitivity and di...

  1. Structure and possible mechanism of the CcbJ methyltransferase from Streptomyces caelestis

    Czech Academy of Sciences Publication Activity Database

    Bauer, J.; Ondrovičová, G.; Najmanová, Lucie; Pevala, V.; Kameník, Zdeněk; Koštan, J.; Janata, Jiří; Kutejová, Eva

    2014-01-01

    Roč. 70, APR 2014 (2014), s. 943-957. ISSN 0907-4449 R&D Projects: GA MŠk(CZ) EE2.3.30.0003; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:61388971 Keywords : CATECHOL-O-METHYLTRANSFERASE * SN2-LIKE TRANSITION-STATE * CRYSTAL-STRUCTURES Subject RIV: CE - Biochemistry Impact factor: 7.232, year: 2013

  2. Selenium induced selenocysteine methyltransferase gene expression and antioxidant enzyme activities in Astragalus chrysochlorus

    OpenAIRE

    Çakir, Özgür; Turgut-Kara, Neslihan; Ari, Şule

    2016-01-01

    Astragalus sp. are used in folk medicine because of their biological activities and are known for the ability to accumulate high levels of selenium (Se). The purpose of this study was to explore gene expression of selenocysteine methyltransferase (SMT), responsible for forming MeSeCys, and activities of ascorbate peroxidase (APX), peroxidase (POX), catalase (CAT) and glutathione reductase (GR) enzymes in callus tissues of Astragalus chrysochlorus growing in different Se-containing media. Quan...

  3. Recombinant rat liver guanidinoacetate methyltransferase: Reactivity and function of sulfhydryl groups

    International Nuclear Information System (INIS)

    Rat liver guanidinoacetate methyltransferase, produced in Escherichia coli by recombinant DNA technique, possesses five cysteine residues per molecule. No disulfide bond is present. Analysis of the chymotryptic peptides derived from the iodo[14C]acetate-modified enzyme shows that Cys-90, Cys-15, Cys-219, and Cys-207 are alkylated by the reagent in order of decreasing reactivity. Incubation of the enzyme with excess 5,5'-dithiobis(2-nitrobenzoate) (DTNB) in the absence and presence of cystamine [2,2'-dithiobis(ethylamine)] causes the appearance of 4 and 5 mol of 2-nitro-5-mercaptobenzoate/mol of enzyme, respectively. Reaction of the methyltransferase with an equimolar amount of DTNB results in an almost quantitative disulfide cross-linking of Cys-15 and Cys-90 with loss of a large portion of the activity. The methyltransferase is completely inactivated by iodoacetate following nonlinear kinetics. Comparison of the extent of inactivation with that of modification of cysteine residues and the experiment with the enzyme whose Cys-15 and Cys-90 are cross-linked suggest that alkylation of Cys-15 and Cys-90 results in a partially active enzyme and that carboxymethylation of Cys-219 completely eliminates enzyme activity. The inactivation of guanidinoacetate methyltransferase by iodoacetate or DTNB is not protected by substrates. Furthermore, disulfide cross-linking of Cys-15 and Cys-90 or carboxymethylation of Cys-219 does not impair the enzyme's capacity to bind S-adenosylmethionine. Thus, these cysteine residues appear to occur outside the active-site region, but their integrity is crucial for the expression of enzyme activity

  4. Vector design for expression of O6-methylguanine-DNA methyltransferase in hematopoietic cells

    OpenAIRE

    Schambach, Axel; Baum, Christopher

    2007-01-01

    Enhancing DNA repair activity of hematopoietic cells by stably integrating gene vectors that express O(6)-methylguanine-DNA-methyltransferase (MGMT) is of major interest for innovative approaches in tumor chemotherapy and for the control of hematopoietic chimerism in the treatment of multiple other acquired or inherited disorders. Crucial determinants of this selection principle are the stringency of treatment with O(6)-alkylating agents and the level of transgenic MGMT expression. Attempts t...

  5. Identification, Characterization, and Ontogenic Study of a Catechol O-methyltransferase from Zebrafish

    OpenAIRE

    Alazizi, Adnan; Liu, Ming-Yih; Williams, Frederick E.; Kurogi, Katsuhisa; Sakakibara, Yoichi; Suiko, Masahito; Liu, Ming-Cheh

    2010-01-01

    To establish the zebrafish as a model for investigating the methylation pathway of drug metabolism, we embarked on the molecular cloning of the zebrafish catechol O-methyltransferase (COMT). By searching the GenBank database, a zebrafish nucleotide sequence encoding a putative COMT was identified. Based on the sequence information, we designed and synthesized oligonucleotides corresponding to its 5’- and 3’-coding regions of this zebrafish COMT. Using the first-strand cDNA reverse-transcribed...

  6. Expression of O6-methylguanine DNA methyltransferase (MGMT) and its clinical significance in gastroenteropancreatic neuroendocrine neoplasm

    OpenAIRE

    Yang, Qiu-chen; Wang, Yu-Hong; Lin, Yuan; Xue, Ling; Chen, Yuan-Jia; Chen, Min-hu; Chen, Jie

    2014-01-01

    O6-methylguanine-DNA methyltransferase (MGMT) is a widespread DNA repair enzyme defending against mutation caused by guanine O6-alkylating agents. Until now, we know only little about the expression of MGMT in gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN). To study the expression of MGMT and its clinical significance in GEP-NEN, 174 specimens of GEP-NEN were examined, of which 152 specimens came from The First Affiliated Hospital, Sun Yat-sen University during October 1995 to Novem...

  7. Survival and tumorigenesis in O6-methylguanine DNA methyltransferase-deficient mice following cyclophosphamide exposure

    OpenAIRE

    Nagasubramanian, Ramamoorthy; Hansen, Ryan J.; Delaney, Shannon M.; Cherian, Mathew M.; Samson, Leona D.; Kogan, Scott C.; Dolan, M. Eileen

    2008-01-01

    O6-methylguanine DNA methyltransferase (MGMT) deficiency is associated with an increased susceptibility to alkylating agent toxicity. To understand the contribution of MGMT in protecting against cyclophosphamide (CP)-induced toxicity, mutagenesis and tumorigenesis, we compared the biological effects of this agent in transgenic Mgmt knockout and wild-type mice. In addition, neurofibromin (Nf1)+/− background was used to increase the likelihood of CP-induced tumorigenesis. Cohorts of Mgmt-profic...

  8. Somatic mutations in glioblastoma are associated with methylguanine-DNA methyltransferase methylation

    OpenAIRE

    McDonald, Kerrie L.; Tabone, Tania; Nowak, Anna K; Erber, Wendy N.

    2015-01-01

    The high level of methylguanine-DNA methyltransferase (MGMT) in glioblastoma is responsible for resistance to alkylating agents, such as temozolomide (TMZ). In glioblastomas with a methylated MGMT promoter, MGMT deficiency is presumed, resulting in an enhanced effect of TMZ. The aim of the present study was to investigate whether genomic alterations work synergistically with MGMT methylation status and contribute to the response to treatment and overall prognosis in glioblastoma. The current ...

  9. Deficiency of Glycine N-Methyltransferase Aggravates Atherosclerosis in Apolipoprotein E–Null Mice

    OpenAIRE

    Chen, Chien-Yu; Ching, Li-Chieh; Liao, Yi-Jen; Yu, Yuan-Bin; Tsou, Chia-Yuan; Shyue, Song-Kun; Chen, Yi-Ming Arthur; Lee, Tzong-Shyuan

    2012-01-01

    The mechanism underlying the dysregulation of cholesterol metabolism and inflammation in atherogenesis is not understood fully. Glycine N-methyltransferase (GNMT) has been implicated in hepatic lipid metabolism and the pathogenesis of liver diseases. However, little is known about the significance of GNMT in atherosclerosis. We showed the predominant expression of GNMT in foamy macrophages of mouse atherosclerotic aortas. Genetic deletion of GNMT exacerbated the hyperlipidemia, inflammation a...

  10. Nicotinamide N-methyltransferase knockdown protects against diet-induced obesity

    OpenAIRE

    Kraus, Daniel; Yang, Qin; Kong, Dong; Banks, Alexander S.; Zhang, Lin; Rodgers, Joseph T; Pirinen, Eija; Pulinilkunnil, Thomas C.; Gong, Fengying; Wang, Ya-chin; Cen, Yana; Sauve, Anthony A.; Asara, John M.; Peroni, Odile D.; Monia, Brett P.

    2014-01-01

    In obesity and type 2 diabetes, Glut4 glucose transporter expression is decreased selectively in adipocytes1. Adipose-specific knockout or overexpression of Glut4 alters systemic insulin sensitivity2. Here we show, using DNA array analyses, that nicotinamide N-methyltransferase (Nnmt) is the most strongly reciprocally regulated gene when comparing gene expression in white adipose tissue (WAT) from adipose-specific Glut4-knockout or adipose-specific Glut4-overexpressing mice with their respect...

  11. Homocysteine Induces Collagen I Expression by Downregulating Histone Methyltransferase G9a

    OpenAIRE

    Lei, Wenjing; Long, Yanjun; LI, Shuang; Liu, Ze; Zhu, Fengxin; Hou, Fan Fan; Nie, Jing

    2015-01-01

    Hyperhomocysteinemia (HHcy) leads to several clinical manifestations including hepatic fibrosis. Excess deposition of extracellular matrix (ECM) components including collagen is the eponymous lesion of liver fibrosis. In this study, we demonstrated that elevated concentration of Hcy induced the expression of collagen type I in cultured human liver cells as well as in liver tissue of HHcy mice. Meanwhile, Hcy inhibited the expression of histone methyltransferase G9a. Mechanistically, silencing...

  12. Histone H3K9 methyltransferase G9a represses PPARγ expression and adipogenesis

    OpenAIRE

    Wang, Lifeng; Xu, Shiliyang; Lee, Ji-Eun; Baldridge, Anne; Grullon, Sean; Peng, Weiqun; Ge, Kai

    2012-01-01

    PPARγ promotes adipogenesis while Wnt proteins inhibit adipogenesis. However, the mechanisms that control expression of these positive and negative master regulators of adipogenesis remain incompletely understood. By genome-wide histone methylation profiling in preadipocytes, we find that among gene loci encoding adipogenesis regulators, histone methyltransferase (HMT) G9a-mediated repressive epigenetic mark H3K9me2 is selectively enriched on the entire PPARγ locus. H3K9me2 and G9a levels dec...

  13. HISTONE H3 LYSINE 9 METHYLTRANSFERASE G9a IS A TRANSCRIPTIONAL COACTIVATOR FOR NUCLEAR RECEPTORS*

    OpenAIRE

    David Y. Lee; Northrop, Jeffrey P.; Kuo, Min-Hao; Stallcup, Michael R.

    2006-01-01

    Methylation of Lys-9 of histone H3 has been associated with repression of transcription. G9a is a histone H3 Lys-9 methyltransferase localized in euchromatin and acts as a corepressor for specific transcription factors. Here we demonstrate that G9a also functions as a coactivator for nuclear receptors, cooperating synergistically with nuclear receptor coactivators GRIP1, CARM1, and p300 in transient transfection assays. This synergy depends strongly on the arginine-specific protein methyltran...

  14. The methyltransferase G9a regulates HoxA9-dependent transcription in AML

    OpenAIRE

    Lehnertz, Bernhard; Pabst, Caroline; Su, Le; Miller, Michelle; Liu, Feng; Yi, Lin; Zhang, Regan; Krosl, Jana; Yung, Eric; Kirschner, Jeanette; Rosten, Patty; Underhill, T. Michael; Jin, Jian; Hébert, Josée; Sauvageau, Guy

    2014-01-01

    Lehnertz et al. identify the histone methyltransferase G9a as a selective regulator of fast proliferating myeloid progenitors with no discernible function in hematopoietic stem cells. Loss of G9a significantly delays disease progression and reduces leukemia stem cell frequency in mouse models of acute myeloid leukemia. G9a interacts with the leukemogenic transcription factor HoxA9 and regulates HoxA9-dependent transcription. These results highlight G9a inhibition as a means to counteract the ...

  15. CCAAT displacement protein/cut homolog recruits G9a histone lysine methyltransferase to repress transcription

    OpenAIRE

    Nishio, Hitomi; Walsh, Martin J.

    2004-01-01

    CCAAT displacement protein/cut homolog (CDP/cut) is a highly conserved homeodomain protein that contains three cut repeat sequences. CDP/cut is a transcriptional factor for many diverse cellular and viral genes that are involved in most cellular processes, including differentiation, development, and proliferation. Here, we report that CDP/cut interacts with a histone lysine methyltransferase (HKMT), G9a, in vivo and in vitro. The deletion of the cut repeats within CDP/cut abrogates the intera...

  16. Negative Regulation of JAK2 by H3K9 Methyltransferase G9a in Leukemia

    OpenAIRE

    Son, Hye-Ju; Kim, Ji-Young; Hahn, Yoonsoo; Seo, Sang-Beom

    2012-01-01

    Histone methylation at specific lysine residues is a crucial regulatory process in transcriptional regulation. Using chromatin immunoprecipitation with microarray technology (ChIP-chip) analysis, we found that the H3K9-me2 target gene JAK2 was an important factor during differentiation of the HL-60 promyelocytic leukemia cell line by all-trans-retinoic acid (ATRA) treatment. Here, we report that the H3K9 methyltransferase G9a negatively regulated JAK2 transcription in histone methyltransferas...

  17. Three-dimensional culture sensitizes epithelial ovarian cancer cells to EZH2 methyltransferase inhibition

    Science.gov (United States)

    Amatangelo, Michael D.; Garipov, Azat; Li, Hua; Conejo-Garcia, Jose R.; Speicher, David W.; Zhang, Rugang

    2013-01-01

    Inhibitors of EZH2 methyltransferase activity have been demonstrated to selectively suppress the growth of diffused large B cell lymphoma (DLBCL) cells with gain-of-function mutations in EZH2, while exhibiting very limited effects on the growth of DLBCL cells with wild-type EZH2. Given that EZH2 is often overexpressed but not mutated in solid tumors, it is important to investigate the determinants of sensitivity of solid tumor cells to EZH2 inhibitors. In the current study, we show that three-dimensional (3D) culture of epithelial ovarian cancer (EOC) cells that overexpress EZH2 sensitizes these cells to EZH2 methyltransferase inhibition. Treatment of EOC cells with GSK343, a specific inhibitor of EZH2 methyltransferase, decreases the level of H3K27Me3, the product of EZH2’s enzymatic activity. However, GSK343 exhibited limited effects on the growth of EOC cells in conventional two-dimensional (2D) culture. In contrast, GSK343 significantly suppressed the growth of EOC cells cultured in 3D matrigel extracellular matrix (ECM), which more closely mimics the tumor microenvironment in vivo. Notably, GSK343 induces apoptosis of EOC cells in 3D but not 2D culture. In addition, GSK343 significantly inhibited the invasion of EOC cells. In summary, we show that the 3D ECM sensitizes EOC cells to EZH2 methyltransferase inhibition, which suppresses cell growth, induces apoptosis and inhibits invasion. Our findings imply that in EZH2 wild-type solid tumors, the ECM tumor microenvironment plays an important role in determining sensitivity to EZH2 inhibition and suggest that targeting the ECM represents a novel strategy for enhancing EZH2 inhibitor efficacy. PMID:23759589

  18. Kupplung der DNA-Methyltransferase M.SssI mit Triplehelix-bildenden Oligodesoxynucleotiden

    OpenAIRE

    Monami, Amélie Joséphine

    2007-01-01

    DNA modifying enzymes, like DNA methyltransferases (DNA MTases) and restriction endonucleases (REases), could in principle be used to map or manipulate genomes. However, these enzymes are inappropriate for that purpose because of their generally short recognition sequences. An increase of their sequence specificity would therefore be desirable. One strategy to produce megaspecific DNA modifying enzymes is to couple them with triple helix forming oligodeoxynucleotide (TFO). These additional DN...

  19. Catecholamine-o-methyltransferase polymorphisms are associated with postoperative pain intensity.

    LENUS (Irish Health Repository)

    Lee, Peter J

    2011-02-01

    single nucleotide polymorphisms (SNPs) in the genes for catecholamine-O-methyltransferase (COMT), μ-opioid receptor and GTP cyclohydrolase (GCH1) have been linked to acute and chronic pain states. COMT polymorphisms are associated with experimental pain sensitivity and a chronic pain state. No such association has been identified perioperatively. We carried out a prospective observational clinical trial to examine associations between these parameters and the development of postoperative pain in patients undergoing third molar (M3) extraction.

  20. Identification and Characterization of a Highly Conserved Crenarchaeal Protein Lysine Methyltransferase with Broad Substrate Specificity

    OpenAIRE

    Chu, Yindi; Zhang, Zhenfeng; Wang, Qian; Luo, Yuanming; Huang, Li

    2012-01-01

    Protein lysine methylation occurs extensively in the Crenarchaeota, a major kingdom in the Archaea. However, the enzymes responsible for this type of posttranslational modification have not been found. Here we report the identification and characterization of the first crenarchaeal protein lysine methyltransferase, designated aKMT, from the hyperthermophilic crenarchaeon Sulfolobus islandicus. The enzyme was capable of transferring methyl groups to selected lysine residues in a substrate prot...

  1. Genetic, physiological and biochemical characterization of multiple methanol methyltransferase isozymes in Methanosarcina acetivorans C2A.

    Science.gov (United States)

    Pritchett, Matthew A; Metcalf, William W

    2005-06-01

    Biochemical evidence suggests that methanol catabolism in Methanosarcina species requires the concerted effort of methanol:5-hydroxybenzimidazolylcobamide methyltransferase (MtaB), a corrinoid-containing methyl-accepting protein (MtaC) and Co-methyl-5-hydroxybenzimidazolylcobamide:2-mercapto-ethanesulphonic acid methyltransferase (MtaA). Here we show that Methanosarcina acetivorans possesses three operons encoding putative methanol-specific MtaB and corrinoid proteins: mtaCB1, mtaCB2 and mtaCB3. Deletion mutants lacking the three operons, in all possible combinations, were constructed and characterized. Strains deleted for any two of the operons grew on methanol, whereas strains lacking all three did not. Therefore, each operon encodes a bona fide methanol-utilizing MtaB/corrinoid protein pair. Most of the mutants were similar to the wild-type strain, with the exception of the DeltamtaCB1 DeltamtaCB2 double mutant, which grew more slowly and had reduced cell yields on methanol medium. However, all mutants displayed significantly longer lag times when switching from growth on trimethylamine to growth on methanol. This indicates that all three operons are required for wild-type growth on methanol and suggests that each operon has a distinct role in the metabolism of this substrate. The combined methanol:CoM methyltransferase activity of strains carrying only mtaCB1 was twofold higher than strains carrying only mtaCB2 and fourfold higher than strains carrying only mtaCB3. Interestingly, the presence of the mtaCB2 and mtaCB3 operons, in addition to the mtaCB1 operon, did not increase the overall methyltransferase activity, suggesting that these strains may be limited by MtaA availability. All deletion mutants were unaffected with respect to growth on trimethylamine and acetate corroborating biochemical evidence indicating that each methanogenic substrate has specific methyltransfer enzymes. PMID:15882413

  2. The histone methyltransferase and putative oncoprotein MMSET is overexpressed in a large variety of human tumors

    DEFF Research Database (Denmark)

    Hudlebusch, Heidi Rye; Santoni-Rugiu, Eric; Simon, Ronald;

    2011-01-01

    Multiple myeloma SET (Suppressor of variegation, Enhancer of zeste, and Trithorax) domain (MMSET) is a histone lysine methyltransferase deregulated in a subgroup of multiple myelomas with the t(4;14)(p16;q32) translocation and poor prognosis. With the aim of understanding, if MMSET can be involve...... in other types of cancer we investigated the expression of MMSET protein in different types of human tumors....

  3. Is catechol-o-methyltransferase gene polymorphism a risk factor in the development of premenstrual syndrome?

    OpenAIRE

    Deveci, Esma Ozturk; Incebiyik, Adnan; Selek, Salih; Camuzcuoglu, Aysun; Hilali, Nese Gul; Camuzcuoglu, Hakan; Erdal, Mehmet Emin; Vural, Mehmet

    2014-01-01

    Objective The objective of this study was to investigate whether there was a correlation between catechol-o-methyltransferase (COMT) gene polymorphism, which is believed to play a role in the etiology of psychotic disorders, and premenstrual syndrome (PMS). Methods Fifty-three women with regular menstrual cycles, aged between 18 and 46 years and diagnosed with PMS according to the American Congress of Obstetrics and Gynecology criteria were included in this study as the study group, and 53 he...

  4. Sensorimotor gating of schizophrenia patients depends on catechol O-methyltransferase Val158Met polymorphism

    OpenAIRE

    Quednow, B B; Wagner, M.; Mössner, R; Maier, W; Kühn, K U

    2010-01-01

    It has been recently shown that Catechol O-methyltransferase (COMT) Val(158)Met polymorphism strongly influences prepulse inhibition (PPI) of the acoustic startle response (ASR) in healthy human volunteers. Given that schizophrenia patients exhibit impairment in PPI and that COMT is a putative susceptibility gene for schizophrenia, we investigated the impact of the COMT Val(158)Met polymorphisms on PPI in schizophrenic inpatients. We analyzed COMT Val(158)Met polymorphisms and assessed startl...

  5. Genetic Contribution of Catechol-O-methyltransferase Polymorphism in Patients with Migraine without Aura

    OpenAIRE

    Park, Jeong Wook; Lee, Kwang Soo; Kim, Joong Seok; Kim, Yeong In; Shin, Hae Eun

    2007-01-01

    Background Recent genetic association studies have investigated the possible genetic role of the dopaminergic system in migraine. Catechol-O-methyltransferase (COMT) is an enzyme that plays a crucial role in the metabolism of dopamine and its genetic polymorphism is associated with three- to fourfold variation of enzymatic activity. Objectives The objective of this study was to elucidate the role of the COMT polymorphism in the genetic susceptibility to migraine and its phenotypic expression ...

  6. Polymorphism of the catechol-O-methyltransferase gene in Han Chinese patients with psoriasis vulgaris

    OpenAIRE

    Lin Gao; Hong Li; Kai Li; Zhu Shen; Ling Liu; Chunying Li; Zhengdong Zhang; Yufeng Liu

    2009-01-01

    Psoriasis vulgaris is defined by a series of linked cellular changes in the skin: hyperplasia of epidermal keratinocytes, vascular hyperplasia and ectasia, and infiltration of T lymphocytes, neutrophils and other types of leukocytes in the affected skin. Catechol-O-methyltransferase ( COMT ) 158 polymorphism can reduce the activity of the COMT enzyme that may trigger defective differentiation of keratinocytes in psoriasis. Immunocytes can degrade and inactivate catecholamines via monamine oxi...

  7. Catechol-O-methyltransferase (COMT) influences the connectivity of the prefrontal cortex at rest

    OpenAIRE

    Tunbridge, Elizabeth M.; Farrell, Sarah M.; Harrison, Paul J; Mackay, Clare E

    2013-01-01

    Catechol-O-methyltransferase (COMT) modulates dopamine in the prefrontal cortex (PFC) and influences PFC dopamine-dependent cognitive task performance. A human COMT polymorphism (Val158Met) alters enzyme activity and is associated with both the activation and functional connectivity of the PFC during task performance, particularly working memory. Here, we used functional magnetic resonance imaging and a data-driven, independent components analysis (ICA) approach to compare resting state funct...

  8. Polymorphism of the catechol-O-methyltransferase gene in Han Chinese patients with psoriasis vulgaris

    OpenAIRE

    Gao, Lin; Li, Hong; Li, Kai; Shen, Zhu; Liu, Ling; Li, Chunying; Zhang, Zhengdong; Liu, Yufeng

    2009-01-01

    Psoriasis vulgaris is defined by a series of linked cellular changes in the skin: hyperplasia of epidermal keratinocytes, vascular hyperplasia and ectasia, and infiltration of T lymphocytes, neutrophils and other types of leukocytes in the affected skin. Catechol-O-methyltransferase (COMT) 158 polymorphism can reduce the activity of the COMT enzyme that may trigger defective differentiation of keratinocytes in psoriasis. Immunocytes can degrade and inactivate catecholamines via monamine oxida...

  9. Towards more specific O6-Methylguanine-DNA methyltransferase (MGMT) inactivators

    OpenAIRE

    CORDEIRO MACHADO, ALESSANDRA; MC MURRY, THOMAS; Rozas, Isabel

    2011-01-01

    PUBLISHED Searching for a novel family of inactivators of the human DNA repair protein O6?methylguanine?DNA methyltransferase (MGMT) which is known to bind to the DNA minor groove, we have computationally modelled and synthesised two series of 2?amino?6?aryloxy?5?nitropyrimidines with morpholino or aminodiaryl substituents (potential minor groove binders) at the 4?position. Synthesis of these compounds was achieved by successive substitution of each of the two Cl atoms of 2?amino?4,6?dichl...

  10. miR-221/222 Target the DNA Methyltransferase MGMT in Glioma Cells

    OpenAIRE

    Cristina Quintavalle; Davide Mangani; Giuseppina Roscigno; Giulia Romano; Angel Diaz-Lagares; Margherita Iaboni; Elvira Donnarumma; Danilo Fiore; Pasqualino De Marinis; Ylermi Soini; Manel Esteller; Gerolama Condorelli

    2013-01-01

    Glioblastoma multiforme (GBM) is one of the most deadly types of cancer. To date, the best clinical approach for treatment is based on administration of temozolomide (TMZ) in combination with radiotherapy. Much evidence suggests that the intracellular level of the alkylating enzyme O(6)-methylguanine-DNA methyltransferase (MGMT) impacts response to TMZ in GBM patients. MGMT expression is regulated by the methylation of its promoter. However, evidence indicates that this is not the only regula...

  11. O6-methylguanine-DNA methyltransferase (MGMT): can function explain a suicidal mechanism?

    OpenAIRE

    Gouws, Chrisna; Pretorius, Petrus Jacobus

    2011-01-01

    Why does O6-methylguanine-DNA methyltransferase (MGMT), an indispensable DNA repair enzyme, have a mechanism which seems to run counter to its importance? This enzyme is key to the removal of detrimental alkyl adducts from guanine bases. Although the mechanism is well known, an unusual feature surrounds its mode of action, which is its so-called suicidal endpoint. In addition, induction of MGMT is highly variable and its kinetics is atypical. These features raise some questions on the seeming...

  12. Arsenic (+3 oxidation state) methyltransferase and the inorganic arsenic methylation phenotype

    International Nuclear Information System (INIS)

    Inorganic arsenic is enzymatically methylated; hence, its ingestion results in exposure to the parent compound and various methylated arsenicals. Both experimental and epidemiological evidences suggest that some of the adverse health effects associated with chronic exposure to inorganic arsenic may be mediated by these methylated metabolites. If i As methylation is an activation process, then the phenotype for inorganic arsenic methylation may determine risk associated with exposure to this metalloid. We examined inorganic arsenic methylation phenotypes and arsenic (+3 oxidation state) methyltransferase genotypes in four species: three that methylate inorganic arsenic (human (Homo sapiens), rat (Rattus norwegicus), and mouse (Mus musculus)) and one that does not methylate inorganic arsenic (chimpanzee, Pan troglodytes). The predicted protein products from arsenic (+3 oxidation state) methyltransferase are similar in size for rat (369 amino acid residues), mouse (376 residues), and human (375 residues). By comparison, a 275-nucleotide deletion beginning at nucleotide 612 in the chimpanzee gene sequence causes a frameshift that leads to a nonsense mutation for a premature stop codon after amino acid 205. The null phenotype for inorganic arsenic methylation in the chimpanzee is likely due to the deletion in the gene for arsenic (+3 oxidation state) methyltransferase that yields an inactive truncated protein. This lineage-specific loss of function caused by the deletion event must have occurred in the Pan lineage after Homo-Pan divergence about 5 million years ago

  13. Chromatin Targeting of de Novo DNA Methyltransferases by the PWWP Domain

    Institute of Scientific and Technical Information of China (English)

    Ying-ZiGe; Min-TiePu; HumairaGowher; Hai-PingWu; Jian-PingDing; AlbertJeltsch; Guo-LiangXu

    2005-01-01

    DNA methylation patterns of mammalian genomes are generated in gametogenesis and early embryonic development. Two de novo DNA methyltransferases, Dnmt3a and Dnmt3b, are responsible for the process. Both en-zymes contain a long N-terminal regulatory region linked to a conserved C-terminal domain responsible forthe catalytic activity. Although a PWWP domain in the N-terminal region has been shown to bind DNA in vitro, it is unclear how the DNA methyltransferases access their substrate in chromatin in vivo. We show here that the two proteins are associated with chromatin including mitotic chromosomes in mammalian cells, and the PWWP domain is essential for the chromatin targeting of the enzymes. The functional significance of PWWPmediated chromatin targeting is suggested by the fact that a missense mutation in this domain of human DNMT3B causes immunodeficiency, centromeric heterochromatin instability, facial anomalies (ICF) syndrome, which is characterized by loss of methylation insatellite DNA, pericentromeric instability, and immunodeficiency. We demonstrate that the mutant protein completely loses its chromatin targeting capacity. Our data establish the PWWP domain as a novel chromatin/chromosome-targeting module and suggest that the PWWP-mediated chromatin association is essential for the function of the de novo methyltransferases during development.

  14. Transcriptome profiling of Set5 and Set1 methyltransferases: Tools for visualization of gene expression

    Directory of Open Access Journals (Sweden)

    Glòria Mas Martín

    2014-12-01

    Full Text Available Cells regulate transcription by coordinating the activities of multiple histone modifying complexes. We recently identified the yeast histone H4 methyltransferase Set5 and discovered functional overlap with the histone H3 methyltransferase Set1 in gene expression. Specifically, using next-generation RNA sequencing (RNA-Seq, we found that Set5 and Set1 function synergistically to regulate specific transcriptional programs at subtelomeres and transposable elements. Here we provide a comprehensive description of the methodology and analysis tools corresponding to the data deposited in NCBI's Gene Expression Omnibus (GEO under the accession number GSE52086. This data complements the experimental methods described in Mas Martín G et al. (2014 and provides the means to explore the cooperative functions of histone H3 and H4 methyltransferases in the regulation of transcription. Furthermore, a fully annotated R code is included to enable researchers to use the following computational tools: comparison of significant differential expression (SDE profiles; gene ontology enrichment of SDE; and enrichment of SDE relative to chromosomal features, such as centromeres, telomeres, and transposable elements. Overall, we present a bioinformatics platform that can be generally implemented for similar analyses with different datasets and in different organisms.

  15. Rapid restriction enzyme free detection of DNA methyltransferase activity based on DNA-templated silver nanoclusters.

    Science.gov (United States)

    Kermani, Hanie Ahmadzade; Hosseini, Morteza; Dadmehr, Mehdi; Ganjali, Mohammad Reza

    2016-06-01

    DNA methylation has significant roles in gene regulation. DNA methyltransferase (MTase) enzyme characterizes DNA methylation and also induces an aberrant methylation pattern that is related to many diseases, especially cancers. Thus, it is required to develop a method to detect the DNA MTase activity. In this study, we developed a new sensitive and reliable method for methyltransferase activity assay by employing DNA-templated silver nanoclusters (DNA/Ag NCs) without using restriction enzymes. The Ag NCs have been utilized for the determination of M.SssI MTase activity and its inhibition. We designed an oligonucleotide probe which contained an inserted six-cytosine loop as Ag NCs formation template. The changes in fluorescence intensity were monitored to quantify the M.SssI activity. The fluorescence spectra showed a linear decrease in the range of 0.4 to 20 U/ml with a detection limit of 0.1 U/ml, which was significant compared with previous reports. The proposed method was applied successfully for demonstrating the Gentamicin effect as MTase inhibitor. The proposed method showed convenient reproducibility and sensitivity indicating its potential for the determination of methyltransferase activity. PMID:27052776

  16. miR-29 Represses the Activities of DNA Methyltransferases and DNA Demethylases

    Directory of Open Access Journals (Sweden)

    Izuho Hatada

    2013-07-01

    Full Text Available Members of the microRNA-29 (miR-29 family directly target the DNA methyltransferases, DNMT3A and DNMT3B. Disturbances in the expression levels of miR-29 have been linked to tumorigenesis and tumor aggressiveness. Members of the miR-29 family are currently thought to repress DNA methylation and suppress tumorigenesis by protecting against de novo methylation. Here, we report that members of the miR-29 family repress the activities of DNA methyltransferases and DNA demethylases, which have opposing roles in control of DNA methylation status. Members of the miR-29 family directly inhibited DNA methyltransferases and two major factors involved in DNA demethylation, namely tet methylcytosine dioxygenase 1 (TET1 and thymine DNA glycosylase (TDG. Overexpression of miR-29 upregulated the global DNA methylation level in some cancer cells and downregulated DNA methylation in other cancer cells, suggesting that miR-29 suppresses tumorigenesis by protecting against changes in the existing DNA methylation status rather than by preventing de novo methylation of DNA.

  17. Purification of homologous protein carboxyl methyltransferase isozymes from human and bovine erythrocytes

    International Nuclear Information System (INIS)

    The authors have purified the two major isozymes of the L-isoaspartyl/D-aspartyl protein methyltransferase from both human and bovine erythrocytes. These four enzymes all have polypeptide molecular weights of approximately 26,500 and appear to be monomers in solution. Each of these enzymes cross-reacts with antibodies directed against protein carboxyl methyltransferase I from bovine brain. Their structures also appear to be similar when analyzed by dodecyl sulfate gel electrophoresis for the large fragments produced by digestion with Staphylococcus aureus protease V8 or when analyzed by high-performance liquid chromatography (HPLC) for tryptic peptides. The structural relatedness of these enzymes was confirmed by sequence analysis of a total of 433 residues in 32 tryptic fragments of the human erythrocyte isozymes I and II and of the bovine erythrocyte isozyme II. They found sequence identity or probable identity in 111 out of 112 residues when they compared the human isozymes I and II and identities in 127 out of 134 residues when the human and bovine isozymes II were compared. These results suggest that the erythrocyte isozymes from both organisms may have nearly identical structures and confirm the similarities in the function of these methyltransferases that have been previously demonstrated

  18. Preliminary characterization of (nucleoside-2′-O-)-methyltransferase crystals from Meaban and Yokose flaviviruses

    Energy Technology Data Exchange (ETDEWEB)

    Mastrangelo, Eloise; Bollati, Michela; Milani, Mario [Department of Biomolecular Sciences and Biotechnology, CNR-INFM, University of Milano, Via Celoria 26, 20133 Milano (Italy); Lamballeire, Xavier de; Brisbare, Nadege [Unité des Virus Emergents, Faculté de Médecine, 27 Boulevard Jean Moulin, 13005 Marseille (France); Dalle, Karen; Lantez, Violaine; Egloff, Marie-Pierre; Coutard, Bruno; Canard, Bruno [Laboratoire Architecture et Fonction des Macromolécules Biologiques, UMR 6098 CNRS ESIL, Case 932, 163 Avenue de Luminy, 13288 Marseille CEDEX 9 (France); Gould, Ernest; Forrester, Naomi [CEH Oxford, Mansfield Road, Oxford OX1 3SR (United Kingdom); Bolognesi, Martino, E-mail: martino.bolognesi@unimi.it [Department of Biomolecular Sciences and Biotechnology, CNR-INFM, University of Milano, Via Celoria 26, 20133 Milano (Italy)

    2006-08-01

    Two methyltransferases from flaviviruses (Meaban and Yokose viruses) have been overexpressed and crystallized. Diffraction data and characterization of the two crystal forms are presented, together with a preliminary molecular-replacement solution for both enzymes. Viral methyltranferases (MTase) are involved in the third step of the mRNA-capping process, transferring a methyl group from S-adenosyl-l-methionine (SAM) to the capped mRNA. MTases are classified into two groups: (guanine-N7)-methyltransferases (N7MTases), which add a methyl group onto the N7 atom of guanine, and (nucleoside-2′-O-)-methyltransferases (2′OMTases), which add a methyl group to a ribose hydroxyl. The MTases of two flaviviruses, Meaban and Yokose viruses, have been overexpressed, purified and crystallized in complex with SAM. Characterization of the crystals together with details of preliminary X-ray diffraction data collection (at 2.8 and 2.7 Å resolution, respectively) are reported here. The sequence homology relative to Dengue virus 2′OMTase and the structural conservation of specific residues in the putative active sites suggest that both enzymes belong to the 2′OMTase subgroup.

  19. MtrA of the sodium ion pumping methyltransferase binds cobalamin in a unique mode.

    Science.gov (United States)

    Wagner, Tristan; Ermler, Ulrich; Shima, Seigo

    2016-01-01

    In the three domains of life, vitamin B12 (cobalamin) is primarily used in methyltransferase and isomerase reactions. The methyltransferase complex MtrA-H of methanogenic archaea has a key function in energy conservation by catalysing the methyl transfer from methyl-tetrahydromethanopterin to coenzyme M and its coupling with sodium-ion translocation. The cobalamin-binding subunit MtrA is not homologous to any known B12-binding proteins and is proposed as the motor of the sodium-ion pump. Here, we present crystal structures of the soluble domain of the membrane-associated MtrA from Methanocaldococcus jannaschii and the cytoplasmic MtrA homologue/cobalamin complex from Methanothermus fervidus. The MtrA fold corresponds to the Rossmann-type α/β fold, which is also found in many cobalamin-containing proteins. Surprisingly, the cobalamin-binding site of MtrA differed greatly from all the other cobalamin-binding sites. Nevertheless, the hydrogen-bond linkage at the lower axial-ligand site of cobalt was equivalently constructed to that found in other methyltransferases and mutases. A distinct polypeptide segment fixed through the hydrogen-bond linkage in the relaxed Co(III) state might be involved in propagating the energy released upon corrinoid demethylation to the sodium-translocation site by a conformational change. PMID:27324530

  20. The use of biodegradable mulch for tomato and broccoli production: Crop yield and quality, mulch deterioration, and growers' perceptions

    Science.gov (United States)

    Cowan, Jeremy Scott

    Biodegradable mulch may offer the benefits of polyethylene mulch for crop production with the added benefit of biodegradability. Four studies were carried out in Mount Vernon, WA to evaluate biodegradable mulch for tomato (Solanum lycopersicum L.) and broccoli (Brassica oleracea var. italica) production. The first study compared four biodegradable mulch treatments: BioAgri, BioTelo, WeedGuardPlus (cellulose product), and SB-PLA-10/11/12 (experimental, non-woven fabric), to polyethylene mulch and bare ground in high tunnels and in the open field for tomato yield and fruit quality over three growing seasons. Biodegradable plastic films produced yields and fruit quality comparable to polyethylene. Moreover, high tunnels increased total and marketable fruit weight five and eight times, respectively, compared to the open field. The second study quantified relationships among visual assessment parameters and mulch mechanical properties. Visual assessments and mechanical property tests of polyethylene, BioAgri, BioTelo, WeedGuardPlus, and SB-PLA-10/11/12, were made over three growing seasons. Regression analyses found the strongest relationship overall (r2 = 0.41) to be between the percent of initial breaking force in the machine direction and log 10 of percent visual deterioration. However, evaluating mulch products individually and increasing sample frequency are recommended for future research. The third study evaluated three biodegradable mulch products, BioAgri, Crown 1, and SB-PLA-11, after soil-incorporation. The average area of recovered mulch fragments decreased for all mulch products over time. The number of mulch fragments initially increased for all mulch products, with the greatest number of Crown 1 and BioAgri fragments recovered 132 and 299 days after incorporation, respectively. At 397 days after soil-incorporation, the total area of recovered fragments of Crown 1 and BioAgri was 0% and 34% of the theoretical maximum area, respectively. The fourth study

  1. The effect of soil mulching with organic mulches, on weed infestation in broccoli and tomato cultivated under polypropylene fibre, and without a cover

    Directory of Open Access Journals (Sweden)

    Kosterna Edyta

    2014-07-01

    Full Text Available An investigation was done on the effect of different types of organic mulches that were applied in form of straw to the soil mulching process, on the weed infestation, number, and fresh mass of weeds in broccoli cv. Milady F1 (Brassica oleracea L. var. italica Plenck and tomato cv. Polfast F1 (Lycopersicon esculentum Mill. grown under polypropylene fibre as a covering, or grown without a covering. The different types of organic straw mulches were: rye (Secale cereale L., corn (Zea mays L., rape (Brassica napus L. subsp. napus, and buckwheat (Fagopyrum esculentum Moench.. All the organic mulches were applied at a dose of 10 t/ha. The effect of the mulches was compared to a control plot which had no mulch. The type of organic mulch applied to the soil mulching process influenced species composition, number, and fresh mass of weeds. This effect could be the result of the properties of the mulch (colour, structure, etc. or the allelopathic effect on the germination and growth of individual weeds species. Irrespective of the investigated factors, 24 and 25 weeds species, respectively, were observed immediately after cover removal and before broccoli and tomato harvest. In the first date of estimation Chenopodium album L., Echinochloa crus-galli (L. P. Beauv., Fallopia convolvulus (L. Á. Löve, Stellaria media (L. Vill., and Viola tricolor L. dominated, however, before the vegetables were harvested Ch. album, V. tricolor, Veronica arvensis L., and E. crus-galli dominated. An application of polypropylene fibre contributed to an increase in the number and fresh mass of weeds in both vegetables in the first date of estimation (after cover removal. During this period, vegetables cannot compete with weeds. It is important to note, though, that before the vegetables were harvested, a decrease was found in the number and fresh mass of weeds in the covered plots. The most efficient weed limiter, both after cover removal and also before the broccoli and

  2. Effect of Zn on seed germination of broccoli%锌对青花菜种子萌发的影响

    Institute of Scientific and Technical Information of China (English)

    刘玲丽; 颉建明; 张帆; 郁继华

    2014-01-01

    以不同Zn2+浓度(0、0.05、0.5、5、10、50、100、200 mg/L)的ZnSO4溶液处理青花菜种子,研究锌对青花菜种子萌发的影响.结果表明:青花菜种子的发芽指标随着Zn2+浓度的增加均表现出先升高后降低的变化趋势;相对发芽率、相对发芽势、发芽指数均在 Zn2+浓度为100 mg/L 时达到最大;活力指数、胚根长、胚芽长和生物量在10 mg/L时达到最大;当Zn2+浓度大于100 mg/L时,对青花菜种子萌发和生长起到抑制作用.%The seeds of broccoli were treated by zinc sulfate solution of different Zn2+ concentrations(0, 0.05,0.5,5,10,50,100,200 mg/L),thus the effect of Zn on seed germination of broccoli was studied.The results showed that all the values of germination indexes firstly increased and then decreased with the in-creasing of Zn2+ concentration.The relative germination rate,relative germination potential and germina-tion index reached to their maximum values when the concentration of Zn2+ was 100 mg/L,while the seed vigor index,radicle root length,bud root length and biomass were all reached to their maximum values when the concentration of Zn2+ was 10 mg/L.When the concentration of Zn2+ was higher than 100 mg/L, it inhibited the germination and growth of broccoli seeds.

  3. 西兰花种子中萝卜硫素酶解浸提工艺研究%Leaching Process Research of Sulforaphane with Broccoli Seeds

    Institute of Scientific and Technical Information of China (English)

    谢述琼; 何珺; 舒华

    2016-01-01

    西兰花中含有丰富的抗癌物质,对大多数癌症都有较强的作用。本文研究了从西兰花种子中酶解浸提萝卜硫素的工艺,确定其最佳的酶解浸提参数。通过单因素梯度实验研究,采用自身芥子苷酶酶解以及密封浸提方式提取西兰花种子中萝卜硫素,主要对酶解时间,酶解料液比,酶解pH,超声浸提乙醇浓度,超声浸提时间,超声浸提次数几种浸提参数进行研究。结果表明:西兰花种子中萝卜硫素酶解浸提最佳条件为:酶解时间为8 h以上,酶解料液比为12,酶解pH为7,超声浸提溶剂为80%乙醇,超声浸提时间为30 min,超声浸提的次数为2次。西兰花种子中萝卜硫素的酶解浸提工艺参数达到最佳时,其提取率可以大大的提高。%Broccoli contains abundant anti-cancer substances, for most cancer it has a stronger effect. This experiment studied Enzyme solution leaching process of sulforaphane in broccoli seeds, determining the optimum enzymic extraction parameters. Through the single factor gradient experiment research, using own mustard glucoside enzyme digestion and seal extraction method to extract sulforaphane in broccoli seeds, enzymatic hydrolysis time, solid-liquid ratio, acidity leaching, extraction of ethanol concentration, extraction time and ultrasonic extraction times were mainly studied. Results showed that the optimum leaching conditions were as follows: enzymolysis time of 8 h, solid-liquid ratio of 12, leaching pH of 7, extraction solvent of 80% ethanol, extraction time for 30 min, the number of ultrasonic extraction of 2 times. The extraction parameter of sulforaphane gets to the best, its extraction efficiency can be greatly improved.

  4. Broccoli production depending on the seed production system and organic and mineral fertilizer Produção de brócolos em função do sistema de produção da semente e do tipo de substrato

    OpenAIRE

    José Carlos Lopes; Janaína Mauri; Adésio Ferreira; Rodrigo S Alexandre; Allan R de Freitas

    2012-01-01

    The experiment was conducted in a greenhouse at the Universidade Federal do Espirito Santo, in Alegre, Espirito Santo state, Brazil, to evaluate the broccoli production depending on the seed production system and organic and mineral fertilization. Seeds of the broccoli cultivar Ramoso Piracicaba were originated from lots of seed obtained in an organic and conventional production system. The experimental design was of randomized blocks, with four replications in a factorial arrangement combina...

  5. Vigor de sementes de brócolos submetidas a coberturas biodegradáveis e micronutrientes Vigor of broccoli seeds submitted to biodegradable coatings and micronutrients

    Directory of Open Access Journals (Sweden)

    Juliana A. Batista

    2005-07-01

    Full Text Available O desenvolvimento e a aplicação de filmes ou coberturas biodegradáveis na horticultura são técnicas praticadas há vários anos, que visam promover melhoria na qualidade do produto. no presente trabalho desenvolveu-se e caracterizou-se biofilmes de pectina, gelatina e ácidos graxos e verificou-se sua eficiência como cobertura para sementes de brócolos (Brassica oleracea L. var. italica. Os filmes foram caracterizados quanto à solubilidade em água. A contagem e a observação de plantas de brócolos, a partir da germinação de sementes cobertas e não cobertas foi realizada em média a cada três dias, aos 27 dias fêz-se a quantificação da matéria fresca e seca. Os filmes de pectina e ácido esteárico foram 100% solúveis em água, enquanto os elaborados com pectina e gelatina (1/1 foram apenas 18%. A aplicação das coberturas filmogênicas nas sementes de brócolos não afetou a emergência das plantas, demonstrando assim sua potencialidade para uso comercial. Os fertilizantes molibdato de sódio (0,2 mg L-1 e ácido bórico (1mg L-1, aplicados juntamente com os biofilmes, também não afetaram a emergência e o desenvolvimento das plantas.The development and application of biodegradable coatings and films have been used for many years in order to improve quality of the coated or packed product. The development and characterization of pectin and pectin/gelatin-based biofilms with fatty acids was evaluated and verified their efficiency as coatings to broccoli seeds (Brassica oleracea L. var. italica. The solubility in water of the films was determined. The counting and visual observations of the germinated broccoli seeds were done each three days and fresh and dried weight of the plants were determined at the end of the experiment. Pectin-based films with stearic acid were 100% soluble in water and composite films of pectin and gelatin were 18%. The application of the biodegradable coatings on broccoli seeds had no effect on the

  6. Effects of Trichoderma harzianum Rifai over Plasmodiophora brassicae Woronin in broccoli, in Escagüey, municipality of Rangel, Mérida State

    Directory of Open Access Journals (Sweden)

    Mirna Labrador Morales

    2016-03-01

    Full Text Available The effectiveness of Trichoderma harzianum in suppressing clubroot of brassicas, which is caused by Plasmodiophora brassicae, was tested on broccoli (Brassica oleracea var. italica Plenck, in field conditions, in Escagüey, municipality of Rangel, Merida State, Venezuela. The experiment showed that the applications of a biopesticide based on this antagonist had a significant effect on the percentage of health plants and the crop yields, in dependence of the dose used. In addition, the relation benefits/cost was also favorable. These results showed that, for these particular conditions, P. brassicae is an adequate biological alternative to control the clubroot of brassicas, no aggressive to environment and human beings, useful for the transition phase toward a sustainable agriculture, without chemical pesticides.

  7. Green pests and diseases-controlling techniques and efficacy of broccoli%花椰菜病虫害绿色防控技术及其效果

    Institute of Scientific and Technical Information of China (English)

    张明真

    2012-01-01

    福建省云霄县佳州村“佳田农民专业合作社”蔬菜生产基地推广实施花椰菜病虫害绿色防控技术,取得显著成效。该文介绍了绿色防控技术的示范效果及主要措施。%Green pests and diseases-controlling techniques of broccoli were popularized and applied to vegetable production bases of Jia tian farmers' professional co-operatives in Jia zhou village, Yunxiao county, Fujian province. This demonstration obtained significant effect. This paper introduced the demonstration effect and main measures of green controlling techniques.

  8. 青花菜新品种瑞青8号的选育%Breeding of Broccoli New Variety Ruiqing No.8

    Institute of Scientific and Technical Information of China (English)

    秦文斌; 戴忠良; 吴国平; 王建华; 张振超; 潘跃平

    2013-01-01

    瑞青8号是以青花菜细胞质雄性不育材料CMS034为不育源,与优良稳定的青花菜自交系0512经过多代回交转育,育成的不育性稳定、经济性状良好的青花菜细胞质雄性不育系CMS0512,再以雄性不育系CMS0512为母本,优良的青花菜自交系038为父本配制的杂交一代品种.瑞青8号株高60 em左右,株幅65 cm左右,叶色绿,叶面光滑;花球半球形,横径17.5 cm左右,纵径14 cm左右;花球紧密,蕾粒细小,色浓绿,花茎细,不中空,单花球重0.55 kg左右,产量达1400 kg/667 m2左右.属中早熟,定植后72 d左右采收,腋芽2~4个,在主花球采收后,侧花球可连续采收1~2次,植株生长势强、整齐度高、抗逆性强,抗霜霉病、黑腐病.%Cauliflower male sterile line CMS034 used as sterile source was crossed and backcrossed by broccoli inbred line 0512 with excellent stability for several successive generations, and broccoli oytoplasmic male sterile line CMS0512 with stable sterility and good economic traits was bred. Then, CMS0512 (female parent) was hybridized by good broccoli inbred line 038 (male parent) , finally the broccoli hybrid Ruiqing No. 8 was bred. The plant height of Ruiqing No. 8 is about 60 cm, and its plant breadth is about 65 cm with green and glossy leaves. The ball - flower is semi - circular and close with the horizontal diameter of a-bout 17.5 cm and vertical diameter of about 14 cm, and it has small bud granulocyte, dark green color, and fine flower stems without hollow. Its single ball - flower is about 0. 55 kg in weight, and its yield is about 1400 kg/667 m2. Ruiqing No. 8 belongs to middle - early maturing variety, and it can be harvested about 72 d after transplanting. This variety has 2-4 axillary buds. After the harvest of main ball - flower, the lateral ball - flowers can be picked continuously for 1 ~ 2 times. Ruiqing No. 8 has strong growth potential, high uniformity, strong adversity resistance, and resistance to downy mildew and

  9. 青花菜主要农艺性状的相关分析%Correlation Analysis of Main Agronomic Characters of Broccoli (B. oleracea var. italica)

    Institute of Scientific and Technical Information of China (English)

    陆信娟; 杨峰; 樊继德

    2011-01-01

    The 21 agronomic characters of 29 cultivars of broccoli (B. oleracea L. var. italica Planch. ) were evaluated, and the correlations among these agronomic characters were analyzed. The correlation analysis indicated that there were significant correlations (P < 0. 05 ) between 23 pairs of characters and very significant correlations (P < 0. 01 ) between 14 pairs of characters.%对29个青花菜品种的21个农艺性状进行了评价和相关分析.相关分析结果表明: 23对性状之间相关性达到显著水平,14对性状之间相关性达到极显著水平.

  10. 中早熟花椰菜品种对比试验初报%Comparison Test of Early Maturing Broccoli Varieties

    Institute of Scientific and Technical Information of China (English)

    孙国坤; 叶明鑫; 黄永修; 李文北

    2015-01-01

    In this test, Fengtianquansong 65d, Changsheng 65d and other 8 new cauliflower varieties were introduced, with Qingnong 65d as comparison. The results showed that there was strong disease resistance, good quality, and high yield in Changsheng 65d. Therefore, Changsheng 65d could be considered as early maturing broccoli variety and extended at Xiamen area.%本实验引进了丰田全松65d、长胜65d等8个青梗花椰菜新品种,以庆农65d作为对照进行比较试验。结果发现,长胜65d品种抗病性强、品质好、产量高、市场畅销,可作为厦门秋季花椰菜中早熟品种进行推广种植。

  11. Variety of Sulforaphane Content Among 15 Broccoli Sprouts%15个西兰花芽苗菜中萝卜硫素含量差异研究

    Institute of Scientific and Technical Information of China (English)

    高灿红; 林俊城; 徐福乐; 郑金贵

    2012-01-01

    High nutrition, rich in sulforaphane, could prevent and inhibit cancer make broccoli sprouts becoming popular in foreign. Composite design of response surface analysis was taken to optimize the extraction of sulforaphane from broccoli sprouts. Then, the optimization extraction method was used to analyze variation of sulforaphane content among 15 broccoli sprouts. The result showed that optimization of extraction method was transonic processing time of 12 min, pH value of 5.0 and hydrolysis time of 4.3 h. There was a significant deference in content of sulforaphane among 15 germplasms of broccoli sprouts. The highest one was CDBY-10 (506.896 (μg/g) and the lowest one was CDBY-22 (50.416 μg/g). Most of their contents were between 0-200 μg/g, frequency fitting were not well. Anymore, sulforaphane content in most of the germplasms was in line with late-maturing>early-maturing>mid-maturing. CDBY-10, CDBY-20, CDBY-15 needed to be further studied and business developed as high sulforaphane content cultivars.%西兰花芽苗菜营养价值高且富含萝卜硫素,具有预防和抑制癌症的功效.为分析种质资源间萝卜、硫素含量的差异,采用响应面中心复合设计法对超声浸提时间、水解时间和提取液pH条件3个因素进行优化分析,获得最优萝卜硫素提取方法,并用该方法对种质资源中萝卜硫素含量差异分析.结果表明,超声处理时间为12 min、pH条件为5.0、水解时间为4.3 h是提取西兰花芽苗菜萝卜、硫素的最佳条件.利用该方法检测到15个种质资源的芽苗菜中萝卜硫素含量差异显著,其中CDBY-10中含量最高(506.896 μg/g)、CDBY-22中含量最低(50.416 μg/g).15个种质资源中萝卜硫素含量主要分布在0~200 μg/g,频次拟合不显著.多数种质资源萝卜硫素含量符合晚熟>早熟>中熟的规律.CDBY-10、CDBY-20、CDBY-15可作为高萝卜、硫素西兰花芽苗菜进行深入科学研究和商业开发.

  12. The catechol-O-methyltransferase inhibitory potential of Z-vallesiachotamine by in silicoand in vitro approaches

    Directory of Open Access Journals (Sweden)

    Carolina dos Santos Passos

    2015-08-01

    Full Text Available AbstractZ-Vallesiachotamine is a monoterpene indole alkaloid that has a β-N-acrylate group in its structure. This class of compounds has already been described in different Psychotriaspecies. Our research group observed that E/Z-vallesiachotamine exhibits a multifunctional feature, being able to inhibit targets related to neurodegeneration, such as monoamine oxidase A, sirtuins 1 and 2, and butyrylcholinesterase enzymes. Aiming at better characterizing the multifunctional profile of this compound, its effect on cathecol-O-methyltransferase activity was investigated. The cathecol-O-methyltransferase activity was evaluated in vitro by a fluorescence-based method, using S-(5′-adenosyl-l-methionine as methyl donor and aesculetin as substrate. The assay optimization was performed varying the concentrations of methyl donor (S-(5′-adenosyl-l-methionine and enzyme. It was observed that the highest concentrations of both factors (2.25 U of the enzyme and 100 µM of S-(5′-adenosyl-l-methionine afforded the more reproducible results. The in vitro assay demonstrated that Z-vallesiachotamine was able to inhibit the cathecol-O-methyltransferase activity with an IC50 close to 200 µM. Molecular docking studies indicated that Z-vallesiachotamine can bind the catechol pocket of catechol-O-methyltransferase enzyme. The present work demonstrated for the first time the inhibitory properties of Z-vallesiachotamine on cathecol-O-methyltransferase enzyme, affording additional evidence regarding its multifunctional effects in targets related to neurodegenerative diseases.

  13. 青花菜新品种湘绿2号、湘绿3号光合特性比较%Comparison of Photosynthetic Characteristics between New Broccoli Cultivars Xianglv No.2 and Xianglv No.3

    Institute of Scientific and Technical Information of China (English)

    姚丹燕; 吴秋云; 龚意辉; 黄科

    2013-01-01

    The new broccoli cultivars,Xianglv No.2 and Xianglv No.3,were taken as materials and broccoli cultivar Shenglv was taken as control to compare the photosynthetic characteristics among the three broccoli cultivars.The results indicated that the leaf photosynthetic rate showed a positive correlation with yield of broccoli,namely the yield of broccoli was high when leaf photosynthetic rate was high without midday depression.The Xianglv No.3 had the highest activity of nitrate reductase,relatively high photosynthetic efficiency and yield,but its fertilizer tolerance was weak.The Xianglv No.2 had the lowest activity of nitrate reductase,the highest photosynthetic efficiency and yield,its fertilizer tolerance was also strong.The assimilation ability of broccoli to nitrate nitrogen was strong in Xianglv No.3 and weak in Xianglv No.2.The Xianglv No.2 could be extended because its yield is the highest,and it is a crop with high photosynthetic efficiency.%以青花菜新品种湘绿2号、湘绿3号为材料,以青花菜圣绿品种为对照,比较了3种青花菜品种的光合特性.结果表明:青花菜叶片的光合速率与产量呈正相关,叶片光合速率高,其产量也高,且没有午休现象.湘绿3号硝酸还原酶活性最高,光合效率也比较高,产量较高,耐肥性弱;湘绿2号硝酸还原酶活性最低,光合效率最高,产量也最高,耐肥性强.湘绿3号对硝态氮的吸收同化能力强,湘绿2号对硝态氮的吸收同化能力弱.湘绿2号产量最高,是高光效的作物,可以推广种植.

  14. 基于颜色参数变化的青花菜叶绿素含量预测模型%A Prediction Model for Chlorophyll Content in Post-harvest Broccoli Based on Color Parameter Changes

    Institute of Scientific and Technical Information of China (English)

    许凤; 杨震峰; 裴娇艳; 郑永华

    2011-01-01

    为研究青花菜贮藏期间叶绿素含量和颜色变化的关系,通过不同温度贮藏实验建立基于颜色参数-a/b值的叶绿素含量预测模型。将青花菜贮藏在273、278、283、293、303K条件下,测定花蕾-a/b值和叶绿素含量的变化。基于Arrhenius动力学方程分别建立青花菜-a/b值、叶绿素含量与贮藏时间和温度之间的动力学模型,再根据-a/b值与叶绿素含量变化的线性关系,进一步建立基于-a/b值的叶绿素含量预测模型。在275、280、285、295、305K贮藏温度条件下对基于-a/b值的叶绿素含量预测模型进行验证,%In order to explore the relationship between chlorophyll content and color change of post-harvest broccoli during storage,a prediction model for chlorophyll content was established based on color parameter changes.Freshly harvested broccoli was stored at different temperatures(273,278,283,293 K and 303 K),and the color parameter-a/b value and chlorophyll content of broccoli were determined.Kinetic models of-a/b value and chlorophyll content with respect to storage time and temperature were established on the basis of Arrhenius equation.In addition,a prediction model for chlorophyll content based on-a/b value was further developed according to the linear relationship between chlorophyll content and-a/b value.Moreover,the prediction model for chlorophyll content based on-a/b value was validated using broccoli stored at 275,280,285,295 K and 305 K with a relative prediction error of 4.31%.The chlorophyll content of broccoli can be accurately predicted at the storage temperature from 273 K to 305 K using this prediction model.Therefore,the prediction model is acceptable for applying the color parameter-a/b to evaluate broccoli quality.

  15. Dietary Intake of Sulforaphane-Rich Broccoli Sprout Extracts during Juvenile and Adolescence Can Prevent Phencyclidine-Induced Cognitive Deficits at Adulthood.

    Directory of Open Access Journals (Sweden)

    Yumi Shirai

    Full Text Available Oxidative stress and inflammation play a role in cognitive impairment, which is a core symptom of schizophrenia. Furthermore, a hallmark of the pathophysiology of this disease is the dysfunction of cortical inhibitory γ-aminobutyric acid (GABA neurons expressing parvalbumin (PV, which is also involved in cognitive impairment. Sulforaphane (SFN, an isothiocyanate derived from broccoli, is a potent activator of the transcription factor Nrf2, which plays a central role in the inducible expressions of many cytoprotective genes in response to oxidative stress. Keap1 is a cytoplasmic protein that is essential for the regulation of Nrf2 activity. Here, we found that pretreatment with SFN attenuated cognitive deficits, the increase in 8-oxo-dG-positive cells, and the decrease in PV-positive cells in the medial prefrontal cortex and hippocampus after repeated administration of phencyclidine (PCP. Furthermore, PCP-induced cognitive deficits were improved by the subsequent subchronic administration of SFN. Interestingly, the dietary intake of glucoraphanin (a glucosinolate precursor of SFN during the juvenile and adolescence prevented the onset of PCP-induced cognitive deficits as well as the increase in 8-oxo-dG-positive cells and the decrease in PV-positive cells in the brain at adulthood. Moreover, the NRF2 gene and the KEAP1 gene had an epistatic effect on cognitive impairment (e.g., working memory and processing speed in patients with schizophrenia. These findings suggest that SFN may have prophylactic and therapeutic effects on cognitive impairment in schizophrenia. Therefore, the dietary intake of SFN-rich broccoli sprouts during the juvenile and adolescence may prevent the onset of psychosis at adulthood.

  16. Comparison of New Broccoli Cultivars in Jiangsu Area%江苏地区青花菜新品种比较试验

    Institute of Scientific and Technical Information of China (English)

    宋立晓; 曾爱松; 高兵; 严继勇

    2014-01-01

    2011-2012年,在江苏南京以圣绿为对照,对江苏省近年来育成的6个青花菜品种进行比较试验。试验结果表明,苏绿3号、苏绿2号、瑞绿5号和瑞绿6号4个品种单花球质量大,产量高,商品花球合格率较高,抗病性强,营养品质较佳,综合性状较好,可作为保鲜出口或中球内销预选品种在江苏省推广种植,为江苏省选择高产、优质、商品性佳的青花菜新品种提供了基础依据。%We took Shenglv as control cultivar, and carried out the comparative test of six broccoli cultivars from 2011 to 2012. The results showed that the four cultivars Sulv No.3, Sulv No.2, Ruilv No.5 and Ruilv No.6 had outstanding comprehensive characters, and their single flower head weight, yield, commercial qualification rate, resistance to diseases and nutrition quality were better than those of control and the other cultivars, so they were suitable for extension of cultivation in Jiangsu area for fresh-keeping export and domestic sales. Furthermore, this comparative test provided basic references for screening of new broccoli cultivars with high yield, excellent quality and good commercial characters.

  17. Genetic examination of SETD7 and SUV39H1/H2 methyltransferases and the risk of diabetes complications in patients with type 1 diabetes

    DEFF Research Database (Denmark)

    Syreeni, Anna; El-Osta, Assam; Forsblom, Carol;

    2011-01-01

    episodes of hyperglycemia. Epigenetic modifications mediated by histone methyltransferases are associated with gene-activating events that promote enhanced expression of key proinflammatory molecules implicated in vascular injury. In this study, we investigated genetic polymorphisms of the SETD7, SUV39H1......, and SUV39H2 methyltransferases as predictors of risk for micro- and macrovascular complications in type 1 diabetes....

  18. Insights into the structure, function and evolution of the radical-SAM 23S rRNA methyltransferase Cfr that confers antibiotic resistance in bacteria

    DEFF Research Database (Denmark)

    Karminska, K. H.; Purta, E.; Hansen, L .H.; Bujnicki, J. M.; Vester, B.; Long, Katherine

    2010-01-01

    The Cfr methyltransferase confers combined resistance to five classes of antibiotics that bind to the peptidyl tranferase center of bacterial ribosomes by catalyzing methylation of the C-8 position of 23S rRNA nucleotide A2503. The same nucleotide is targeted by the housekeeping methyltransferase...

  19. Epigenetic changes of Arabidopsis genome associated with altered DNA methyltransferase and demethylase expressions after gamma irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Ji Eun; Cho, Eun Ju; Kim, Ji Hong; Chung, Byung Yeoup; Kim, Jin Hong [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

    2012-05-15

    DNA methylation at carbon 5 of cytosines is a hall mark of epigenetic inactivation and heterochromatin in both plants and mammals. In Arabidopsis, DNA methylation has two roles that protect the genome from selfish DNA elements and regulate gene expression. Plant genome has three types of DNA methyltransferase, METHYLTRANSFERASE 1 (MET1), DOMAINREARRANGED METHYLASE (DRM) and CHROMOMETHYLASE 3 (CMT3) that are capable of methylating CG, CHG (where H is A, T, or C) and CHH sites, respectively. MET1 is a maintenance DNA methyltransferase that controls CG methylation. Two members of the DRM family, DRM1 and DRM2, are responsible for de novo methylation of CG, CHG, and CHH sites but show a preference for CHH sites. Finally, CMT3 principally carries out CHG methylation and is involved in both de novo methylation and maintenance. Alternatively, active DNA demethylation may occur through the glycosylase activity by removing the methylcytosines from DNA. It may have essential roles in preventing transcriptional silencing of transgenes and endogenous genes and in activating the expression of imprinted genes. DNA demetylation in Arabidopsis is mediated by the DEMETER (DME) family of bifunctional DNA glycosylase. Three targets of DME are MEA (MEDEA), FWA (FLOWERING WAGENINGEN), and FIS2 (FERTILIZATION INDEPENDENT SEED 2). The DME family contains DEMETER-LIKE 2 (DML2), DML3, and REPRESSOR OF SILENING 1 (ROS1). DNA demetylation by ROS1, DML2, and DML3 protect the hypermethylation of specific genome loci. ROS1 is necessary to suppress the promoter methylation and the silencing of endogenous genes. In contrast, the function of DML2 and DML3 has not been reported. Several recent studies have suggested that epigenetic alterations such as change in DNA methylation and histone modification should be caused in plant genomes upon exposure to ionizing radiation. However, there is a lack of data exploring the underlying mechanisms. Therefore, the present study aims to characterize and

  20. Epigenetic changes of Arabidopsis genome associated with altered DNA methyltransferase and demethylase expressions after gamma irradiation

    International Nuclear Information System (INIS)

    DNA methylation at carbon 5 of cytosines is a hall mark of epigenetic inactivation and heterochromatin in both plants and mammals. In Arabidopsis, DNA methylation has two roles that protect the genome from selfish DNA elements and regulate gene expression. Plant genome has three types of DNA methyltransferase, METHYLTRANSFERASE 1 (MET1), DOMAINREARRANGED METHYLASE (DRM) and CHROMOMETHYLASE 3 (CMT3) that are capable of methylating CG, CHG (where H is A, T, or C) and CHH sites, respectively. MET1 is a maintenance DNA methyltransferase that controls CG methylation. Two members of the DRM family, DRM1 and DRM2, are responsible for de novo methylation of CG, CHG, and CHH sites but show a preference for CHH sites. Finally, CMT3 principally carries out CHG methylation and is involved in both de novo methylation and maintenance. Alternatively, active DNA demethylation may occur through the glycosylase activity by removing the methylcytosines from DNA. It may have essential roles in preventing transcriptional silencing of transgenes and endogenous genes and in activating the expression of imprinted genes. DNA demetylation in Arabidopsis is mediated by the DEMETER (DME) family of bifunctional DNA glycosylase. Three targets of DME are MEA (MEDEA), FWA (FLOWERING WAGENINGEN), and FIS2 (FERTILIZATION INDEPENDENT SEED 2). The DME family contains DEMETER-LIKE 2 (DML2), DML3, and REPRESSOR OF SILENING 1 (ROS1). DNA demetylation by ROS1, DML2, and DML3 protect the hypermethylation of specific genome loci. ROS1 is necessary to suppress the promoter methylation and the silencing of endogenous genes. In contrast, the function of DML2 and DML3 has not been reported. Several recent studies have suggested that epigenetic alterations such as change in DNA methylation and histone modification should be caused in plant genomes upon exposure to ionizing radiation. However, there is a lack of data exploring the underlying mechanisms. Therefore, the present study aims to characterize and

  1. 2'-O methylation of internal adenosine by flavivirus NS5 methyltransferase.

    Directory of Open Access Journals (Sweden)

    Hongping Dong

    Full Text Available RNA modification plays an important role in modulating host-pathogen interaction. Flavivirus NS5 protein encodes N-7 and 2'-O methyltransferase activities that are required for the formation of 5' type I cap (m(7GpppAm of viral RNA genome. Here we reported, for the first time, that flavivirus NS5 has a novel internal RNA methylation activity. Recombinant NS5 proteins of West Nile virus and Dengue virus (serotype 4; DENV-4 specifically methylates polyA, but not polyG, polyC, or polyU, indicating that the methylation occurs at adenosine residue. RNAs with internal adenosines substituted with 2'-O-methyladenosines are not active substrates for internal methylation, whereas RNAs with adenosines substituted with N⁶-methyladenosines can be efficiently methylated, suggesting that the internal methylation occurs at the 2'-OH position of adenosine. Mass spectroscopic analysis further demonstrated that the internal methylation product is 2'-O-methyladenosine. Importantly, genomic RNA purified from DENV virion contains 2'-O-methyladenosine. The 2'-O methylation of internal adenosine does not require specific RNA sequence since recombinant methyltransferase of DENV-4 can efficiently methylate RNAs spanning different regions of viral genome, host ribosomal RNAs, and polyA. Structure-based mutagenesis results indicate that K61-D146-K181-E217 tetrad of DENV-4 methyltransferase forms the active site of internal methylation activity; in addition, distinct residues within the methyl donor (S-adenosyl-L-methionine pocket, GTP pocket, and RNA-binding site are critical for the internal methylation activity. Functional analysis using flavivirus replicon and genome-length RNAs showed that internal methylation attenuated viral RNA translation and replication. Polymerase assay revealed that internal 2'-O-methyladenosine reduces the efficiency of RNA elongation. Collectively, our results demonstrate that flavivirus NS5 performs 2'-O methylation of internal adenosine of

  2. Molecular Evolution of the Substrate Specificity of Chloroplastic Aldolases/Rubisco Lysine Methyltransferases in Plants.

    Science.gov (United States)

    Ma, Sheng; Martin-Laffon, Jacqueline; Mininno, Morgane; Gigarel, Océane; Brugière, Sabine; Bastien, Olivier; Tardif, Marianne; Ravanel, Stéphane; Alban, Claude

    2016-04-01

    Rubisco and fructose-1,6-bisphosphate aldolases (FBAs) are involved in CO2 fixation in chloroplasts. Both enzymes are trimethylated at a specific lysine residue by the chloroplastic protein methyltransferase LSMT. Genes coding LSMT are present in all plant genomes but the methylation status of the substrates varies in a species-specific manner. For example, chloroplastic FBAs are naturally trimethylated in both Pisum sativum and Arabidopsis thaliana, whereas the Rubisco large subunit is trimethylated only in the former species. The in vivo methylation status of aldolases and Rubisco matches the catalytic properties of AtLSMT and PsLSMT, which are able to trimethylate FBAs or FBAs and Rubisco, respectively. Here, we created chimera and site-directed mutants of monofunctional AtLSMT and bifunctional PsLSMT to identify the molecular determinants responsible for substrate specificity. Our results indicate that the His-Ala/Pro-Trp triad located in the central part of LSMT enzymes is the key motif to confer the capacity to trimethylate Rubisco. Two of the critical residues are located on a surface loop outside the methyltransferase catalytic site. We observed a strict correlation between the presence of the triad motif and the in vivo methylation status of Rubisco. The distribution of the motif into a phylogenetic tree further suggests that the ancestral function of LSMT was FBA trimethylation. In a recent event during higher plant evolution, this function evolved in ancestors of Fabaceae, Cucurbitaceae, and Rosaceae to include Rubisco as an additional substrate to the archetypal enzyme. Our study provides insight into mechanisms by which SET-domain protein methyltransferases evolve new substrate specificity. PMID:26785049

  3. Human erythrocyte phenol O-methyltransferase: Radiochemical microassay and biochemical properties

    International Nuclear Information System (INIS)

    A radiochemical microassay for the determination of phenol O-methyltransferase (PMT) activity in human red blood cell membranes has been developed. Acetaminophen was used as the substrate. The apparent Michaelis-Menten Ksub(M) value for acetaminophen was 21.2 X 10-3M. The apparent Ksub(M) value for S-adenosyl-L-methionine, a co-substrate for the reaction, was 4.8 X 10-6 M, and the pH optimum of the reaction was approximately 9.0 with four different buffer systems. Phenol was also tested as a substrate and had an apparent Ksub(M) value of 2.0 X 10-3 M. Human erythrocyte (RBC) membrane PMT activity did not have the biochemical characteristics of catechol O-methyltransferase, another RBC membrane methyltransferase enzyme activity. Blood samples obtained from 212 randomly selected adult white subjects had a mean activity of 134.5 +41.5 pmol of p-acetanisidide formed per mg protein per hour (mean +- S.D.). Activities varied from 44 to 282 units. There were no differences in the mean activities of samples from men and women. Experiments in which mixtures of 'low' and 'high' activity RBC membrane preparations were assayed for PMT provided no evidence that the variations in enzyme activity were due to the presence of endogenous PMT activators or inhibitors. RBC membrane PMT activity in blood from 9 patients with renal failure, a pathological state in which there are elevated circulating levels of phenols, was found to be significantly decreased with average activity of 76.2+-9.7 (mean +- S.E.M., P <0.001). (Auth.)

  4. Arabidopsis DNA methyltransferase AtDNMT2 associates with histone deacetylase AtHD2s activity

    International Nuclear Information System (INIS)

    DNA methyltransferase2 (DNMT2) is always deemed to be enigmatic, because it contains highly conserved DNA methyltransferase motifs but lacks the DNA methylation catalytic capability. Here we show that Arabidopsis DNA methyltransferase2 (AtDNMT2) is localized in nucleus and associates with histone deacetylation. Bimolecular fluorescence complementation and pull-down assays show AtDNMT2 interacts with type-2 histone deacetylases (AtHD2s), a unique type of histone deacetylase family in plants. Through analyzing the expression of AtDNMT2: ss-glucuronidase (GUS) fusion protein, we demonstrate that AtDNMT2 has the ability to repress gene expression at transcription level. Meanwhile, the expression of AtDNMT2 gene is altered in athd2c mutant plants. We propose that AtDNMT2 possibly involves in the activity of histone deacetylation and plant epigenetic regulatory network.

  5. Arabidopsis DNA methyltransferase AtDNMT2 associates with histone deacetylase AtHD2s activity

    Energy Technology Data Exchange (ETDEWEB)

    Song, Yuan [Key Laboratory of Arid and Grassland Agroecology, Ministry of Education, School of Life Science, Lanzhou University, Lanzhou 730000 (China); Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, ON, Canada N5V4T3 (Canada); Wu, Keqiang [Institute of Plant Biology, National Taiwan University, Taipei 106, Taiwan (China); Dhaubhadel, Sangeeta [Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, ON, Canada N5V4T3 (Canada); An, Lizhe, E-mail: lizhean@lzu.edu.cn [Key Laboratory of Arid and Grassland Agroecology, Ministry of Education, School of Life Science, Lanzhou University, Lanzhou 730000 (China); Tian, Lining, E-mail: tianl@agr.gc.ca [Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, ON, Canada N5V4T3 (Canada)

    2010-05-28

    DNA methyltransferase2 (DNMT2) is always deemed to be enigmatic, because it contains highly conserved DNA methyltransferase motifs but lacks the DNA methylation catalytic capability. Here we show that Arabidopsis DNA methyltransferase2 (AtDNMT2) is localized in nucleus and associates with histone deacetylation. Bimolecular fluorescence complementation and pull-down assays show AtDNMT2 interacts with type-2 histone deacetylases (AtHD2s), a unique type of histone deacetylase family in plants. Through analyzing the expression of AtDNMT2: ss-glucuronidase (GUS) fusion protein, we demonstrate that AtDNMT2 has the ability to repress gene expression at transcription level. Meanwhile, the expression of AtDNMT2 gene is altered in athd2c mutant plants. We propose that AtDNMT2 possibly involves in the activity of histone deacetylation and plant epigenetic regulatory network.

  6. Development of fed-batch profiles for efficient biosynthesis of catechol-O-methyltransferase

    OpenAIRE

    G.M. Espírito Santo; A.Q. Pedro; D. Oppolzer; Bonifácio, M J; Queiroz, J. A.; Silva, F.; L.A. Passarinha

    2014-01-01

    Catechol-O-methyltransferase (COMT, EC 2.1.1.6) plays a crucial role in dopamine metabolism which has intimately linked this enzyme to some neurodegenerative diseases, such as Parkinson's disease. In recent years, in the attempt of developing new therapeutic strategies for Parkinson's disease, there has been a growing interest in the search for effective COMT inhibitors. In order to do so, large amounts of COMT in an active form are needed, and the best way to achieve this is by up-scaling it...

  7. Structural Basis for Binding of RNA and Cofactor by a KsgA Methyltransferase

    OpenAIRE

    Tu, Chao; Tropea, Joseph E.; Austin, Brian P; Court, Donald L.; Waugh, David S.; Ji, Xinhua

    2009-01-01

    Among methyltransferases, KsgA and the reaction it catalyzes are conserved throughout evolution. However, the specifics of substrate recognition by the enzyme remain unknown. Here, we report structures of Aquifex aeolicus KsgA, in its ligand-free form, in complex with RNA and in complex with both RNA and S-adenosylhomocysteine (SAH, reaction product of cofactor S-adenosylmethionine), providing the first pieces of structural information on KsgA-RNA and KsgA-SAH interactions. Moreover, the stru...

  8. Inhibition of SUV39H1 Methyltransferase Activity by DBC1*

    OpenAIRE

    Li, Zhenyu; Chen, Lihong; Kabra, Neha; Wang, Chuangui; Fang, Jia; Chen, Jiandong

    2009-01-01

    SUV39H1 is a histone H3K9-specific methyltransferase important for heterochromatin formation, regulation of gene expression, and induction of senescence in premalignant cells. SUV39H1 forms a complex with SirT1, and its activity is stimulated by SirT1 binding. Here we present evidence that the product of the DBC1 (deleted in breast cancer 1) gene disrupts the SUV39H1-SirT1 complex. Furthermore, DBC1 binds to the SUV39H1 catalytic domain and inhibits its ability to ...

  9. Histone methyltransferase Suv39h1 represses MyoD-stimulated myogenic differentiation

    OpenAIRE

    Mal, Asoke K.

    2006-01-01

    Suv39h1 is a histone H3 lysine-9 (H3-K9) specific methyltransferase (HMT) that is associated with gene silencing through chromatin modification. The transition from proliferation into differentiation of muscle cell is accompanied by transcriptional activation of previously silent muscle genes. I report Suv39h1 interaction with myogenic regulator MyoD in proliferating muscle cells and its HMT activity, which is associated with MyoD, diminishes as differentiation proceeds. The Suv39h1–MyoD comp...

  10. The DmtA methyltransferase contributes to Aspergillus flavus conidiation, sclerotial production, aflatoxin biosynthesis and virulence

    OpenAIRE

    Kunlong Yang; Linlin Liang; Fanlei Ran; Yinghang Liu; Zhenguo Li; Huahui Lan; Peili Gao; Zhenhong Zhuang; Feng Zhang; Xinyi Nie; Shimuye Kalayu Yirga; Shihua Wang

    2016-01-01

    DNA methylation is essential for epigenetic regulation of gene transcription and development in many animals, plants and fungi. We investigated whether DNA methylation plays a role in the development and secondary metabolism of Aspergillus flavus, identified the DmtA methyltransferase from A. flavus, and produced a dmtA knock-out mutant by replacing the dmtA coding sequence with the pyrG selectable marker. The A. flavus dmtA null mutant lines produced white fluffy mycelium in liquid medium, a...

  11. Histone methyltransferase G9a contributes to H3K27 methylation in vivo

    Institute of Scientific and Technical Information of China (English)

    Hui Wu; Bing Zhu; Xiuzhen Chen; Jun Xiong; Yingfeng Li; Hong Li; Xiaojun Ding; Sheng Liu; She Chen; Shaorong Gao

    2011-01-01

    @@ Dear Editor, Histone modifications play a vital role in the conformation and function of their associated chromatin templates[1].Histone H3K27 methylation mediated by the PRC2 complex is critical for transcriptional regulation,Polycomb silencing,Drosophila segmentation,mammalian X inactivation and cancer[1].Interestingly,H3K27me1(H3 mono-methylated at residue K27)levels in vivo remain unaffected after PRC2 disruption[2,3],which is an indication for the existence of other contributing histone methyltransferase(s)to H3K27me1.

  12. EmtA, a rRNA methyltransferase conferring high-level evernimicin resistance

    DEFF Research Database (Denmark)

    Mann, P. A.; Xiong, L.; Mankin, A. S.;

    2001-01-01

    Enterococcus faecium strain 9631355 was isolated from animal sources on the basis of its resistance to the growth promotant avilamycin. The strain also exhibited high-level resistance to evernimicin, a drug undergoing evaluation as a therapeutic agent in humans. Ribosomes from strain 9631355...... exhibited a dramatic reduction in evernimicin binding, shown by both cell-free translation assays and direct-binding assays. The resistance determinant was cloned from strain 9631355; sequence alignments suggested it was a methyltransferase and therefore it was designated emtA for evernimicin...

  13. Juvenile hormone acid methyltransferase: A key regulatory enzyme for insect metamorphosis

    OpenAIRE

    Shinoda, Tetsuro; Itoyama, Kyo

    2003-01-01

    Juvenile hormone (JH) acid methyltransferase (JHAMT) is an enzyme that converts JH acids or inactive precursors of JHs to active JHs at the final step of JH biosynthesis pathway in insects. By fluorescent mRNA differential display, we have cloned a cDNA encoding JHAMT from the corpora allata (CA) of the silkworm, Bombyx mori (BmJHAMT). The BmJHAMT cDNA encodes an ORF of 278 aa with a calculated molecular mass of 32,544 Da. The predicted amino acid sequence contains a conserved S-adenosyl-l-me...

  14. Crystal structure of phosphoethanolamine methyltransferase from Plasmodium falciparum in complex with amodiaquine

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Soon Goo; Alpert, Tara D.; Jez, Joseph M. (WU)

    2012-07-17

    Phosphoethanolamine N-methyltransferase (PMT) is essential for phospholipid biogenesis in the malarial parasite Plasmodium falciparum. PfPMT catalyzes the triple methylation of phosphoethanolamine to produce phosphocholine, which is then used for phosphatidylcholine synthesis. Here we describe the 2.0 {angstrom} resolution X-ray crystal structure of PfPMT in complex with amodiaquine. To better characterize inhibition of PfPMT by amodiaquine, we determined the IC{sub 50} values of a series of aminoquinolines using a direct radiochemical assay. Both structural and functional analyses provide a possible approach for the development of new small molecule inhibitors of PfPMT.

  15. O6-Methylguanine-DNA methyltransferase deficiency in developing brain: Implications for brain tumorigenesis

    OpenAIRE

    Bobola, Michael S.; Blank, A.; Berger, Mitchel S.; Silber, John R

    2007-01-01

    The DNA repair protein O6-methylguanine-DNA methyltransferase (MGMT) is a cardinal defense against the mutagenic and carcinogenic effects of alkylating agents. We have reported evidence that absence of detectable MGMT activity (MGMT− phenotype) in human brain is a predisposing factor for primary brain tumors that affects ca. 12% of individuals [J.R. Silber, et. al. Proc. Natl. Acad. Sci. USA 93 (1996) 6941–6946]. We report here that MGMT− phenotype in the brain of children and adults, and the...

  16. Verminderte Expression von O6-Methylguanin-DNA-Methyltransferase bei Gliomen durch Promotormethylierung

    OpenAIRE

    Nickolay, Carla

    2009-01-01

    Pro Jahr erkranken 10 von 100 000 Menschen an einem malignen Hirntumor. Die Prognose ist schlecht. Die Therapie schließt Chirurgie, Bestrahlung und Chemotherapie ein. Das DNA-Reparaturenzym O6-Methylguanin-DNA-Methyltransferase, MGMT, erkennt DNA-Schäden, die durch alkylierende Substanzen wie Temozolomid entstanden sind. Übermäßige Methylierung der Promotorregion von MGMT führt zum Funktionsverlust des Enzyms. Dieser kann zu einer erhöhten Sensibilität für Alkylanzien führen. In der Therapie ...

  17. DNA Methyltransferase Gene dDnmt2 and Longevity of Drosophila

    Institute of Scientific and Technical Information of China (English)

    Meng-JauLin; Lin-YaTang; M.NarsaReddy; C.K.JamesShen

    2005-01-01

    The DNA methylation program of the fruit fly Drosophila melanogaster is carried out by the single DNA methyltransferase gene dDnmt2, the function of which is unknown before. We present evidence that intactness of the gene is required for maintenance of the normal life span of the fruit flies. In contrast, overexpression of dDnmt2 could extend Drosophila life span. The study links the Drosophila DNA methylation program with the small heatshock proteins and longevity/aging and has interesting implication on the eukaryotic DNA methyl-ation programs in general.

  18. Discovery of an in vivo Chemical Probe of the Lysine Methyltransferases G9a and GLP

    OpenAIRE

    Liu, Feng; Barsyte-Lovejoy, Dalia; Li, Fengling; Xiong, Yan; Korboukh, Victoria; Huang, Xi-Ping; Allali-Hassani, Abdellah; Janzen, William P.; Roth, Bryan L.; Frye, Stephen V.; Arrowsmith, Cheryl H.; Brown, Peter J.; Vedadi, Masoud; Jin, Jian

    2013-01-01

    Among epigenetic “writers”, “readers”, and “erasers”, the lysine methyltransferases G9a and GLP, which catalyze mono- and dimethylation of histone H3 lysine 9 (H3K9me2) and non-histone proteins, have been implicated in a variety of human diseases. A “toolkit” of well-characterized chemical probes will allow biological and disease hypotheses concerning these proteins to be tested in cell-based and animal models with high confidence. We previously discovered potent and selective G9a/GLP inhibit...

  19. Protein lysine methyltransferase G9a acts on non-histone targets

    OpenAIRE

    Rathert, Philipp; Dhayalan, Arunkumar; Murakami, Marie; Zhang, Xing; Tamas, Raluca; Jurkowska, Renata; Komatsu, Yasuhiko; Shinkai, Yoichi; Cheng, Xiaodong; Jeltsch, Albert

    2008-01-01

    By methylation of peptide arrays, we determined the specificity profile of the protein methyltransferase G9a. We show that it mostly recognizes an Arg-Lys sequence and that its activity is inhibited by methylation of the arginine residue. Using the specificity profile, we identified new non-histone protein targets of G9a, including CDYL1, WIZ, ACINUS and G9a (automethylation), as well as peptides derived from CSB. We demonstrate potential downstream signaling pathways for methylation of non-h...

  20. A chemical probe selectively inhibits G9a and GLP methyltransferase activity in cells

    OpenAIRE

    Vedadi, Masoud; Barsyte-Lovejoy, Dalia; Liu, Feng; Rival-Gervier, Sylvie; Allali-Hassani, Abdellah; Labrie, Viviane; Wigle, Tim J.; DiMaggio, Peter A.; Wasney, Gregory A.; Siarheyeva, Alena; Dong, Aiping; Tempel, Wolfram; Wang, Sun-Chong; Chen, Xin; Chau, Irene

    2011-01-01

    Protein lysine methyltransferases G9a and GLP modulate the transcriptional repression of a variety of genes via dimethylation of Lys9 on histone H3 (H3K9me2) as well as dimethylation of non-histone targets. Here we report the discovery of UNC0638, an inhibitor of G9a and GLP with excellent potency and selectivity over a wide range of epigenetic and non-epigenetic targets. UNC0638 treatment of a variety of cell lines resulted in lower global H3K9me2 levels, equivalent to levels observed for sm...

  1. Quantitative proteomics reveals direct and indirect alterations in the histone code following methyltransferase knockdown

    OpenAIRE

    Plazas-Mayorca, Mariana D.; Bloom, Joshua S.; Zeissler, Ulrike; LeRoy, Gary; Young, Nicolas L.; DiMaggio, Peter A.; Krugylak, Leonid; Schneider, Robert; Garcia, Benjamin A

    2010-01-01

    Histones are highly conserved proteins that organize cellular DNA. These proteins, especially their N-terminal domains, are adorned with many post-translational modifications (PTMs) such as lysine methylation, which are associated with active or repressed transcriptional states. The lysine methyltransferase G9a and its interaction partner Glp1 can mono- or dimethylate histone H3 on lysine (H3K9me1 or me2); possible cross-talk between these modifications and other PTMs on the same or other his...

  2. A fluorescence resonance energy transfer-based method for histone methyltransferases

    DEFF Research Database (Denmark)

    Devkota, Kanchan; Lohse, Brian; Nyby Jakobsen, Camilla;

    2015-01-01

    A simple dye–quencher fluorescence resonance energy transfer (FRET)-based assay for methyltransferases was developed and used to determine kinetic parameters and inhibitory activity at EHMT1 and EHMT2. Peptides mimicking the truncated histone H3 tail were functionalized in each end with a dye and a...... quencher, respectively. When lysine-9 residues in the peptides were methylated, they were protected from cleavage by endoproteinase–EndoLysC, whereas unmethylated peptides were cleaved, resulting in an increase in fluorescent intensity....

  3. Regioselectivity of catechol O-methyltransferase confers enhancement of catalytic activity

    Science.gov (United States)

    Tsao, Douglas; Liu, Shubin; Dokholyan, Nikolay V.

    2011-04-01

    Catechol O-methyltransferase (COMT) metabolizes catechol moieties by methylating a single hydroxyl group at the meta- or para- hydroxyl position. Hydrophobic amino acids near the active site of COMT influence the regioselectivity of this reaction. Our sequence analysis highlights their importance by showing that these residues are highly conserved throughout evolution. Reaction barriers calculated in the gas phase reveal a lower barrier during methylation at the meta- position, suggesting that the observed meta-regioselectivity of COMT can be attributed to the substrate itself, and that COMT has evolved residues to orient the substrate in a manner that increases the rate of catalysis.

  4. Small RNA-mediated DNA (cytosine-5) methyltransferase 1 inhibition leads to aberrant DNA methylation

    OpenAIRE

    Zhang, Guoqiang; Estève, Pierre-Olivier; Chin, Hang Gyeong; Terragni, Jolyon; Dai, Nan; Corrêa, Ivan R.; Pradhan, Sriharsa

    2015-01-01

    Mammalian cells contain copious amounts of RNA including both coding and noncoding RNA (ncRNA). Generally the ncRNAs function to regulate gene expression at the transcriptional and post-transcriptional level. Among ncRNA, the long ncRNA and small ncRNA can affect histone modification, DNA methylation targeting and gene silencing. Here we show that endogenous DNA methyltransferase 1 (DNMT1) co-purifies with inhibitory ncRNAs. MicroRNAs (miRNAs) bind directly to DNMT1 with high affinity. The bi...

  5. Queen pheromones modulate DNA methyltransferase activity in bee and ant workers.

    Science.gov (United States)

    Holman, Luke; Trontti, Kalevi; Helanterä, Heikki

    2016-01-01

    DNA methylation is emerging as an important regulator of polyphenism in the social insects. Research has concentrated on differences in methylation between queens and workers, though we hypothesized that methylation is involved in mediating other flexible phenotypes, including pheromone-dependent changes in worker behaviour and physiology. Here, we find that exposure to queen pheromone affects the expression of two DNA methyltransferase genes in Apis mellifera honeybees and in two species of Lasius ants, but not in Bombus terrestris bumblebees. These results suggest that queen pheromones influence the worker methylome, pointing to a novel proximate mechanism for these key social signals. PMID:26814223

  6. DNA methyltransferase-dependent transcription of the phage Mu mom gene.

    OpenAIRE

    Hattman, S

    1982-01-01

    The phage Mu mom gene controls an unusual DNA modification. Expression of the mom function requires an active host (dam+) DNA adenine methylase [S-adenosyl-L-methionine:DNA (6-aminopurine)-methyltransferase]; in dam- hosts, Mu development is normal except that the viral DNA does not undergo the mom modification. The present communication compares transcription of the mom gene in dam+ versus dam- cells. 32P-labeled probes were prepared by nick-translation of a purified mom gene-containing rest...

  7. New erythromycin derivatives from Saccharopolyspora erythraea using sugar O-methyltransferases from the spinosyn biosynthetic gene cluster.

    Science.gov (United States)

    Gaisser, S; Lill, R; Wirtz, G; Grolle, F; Staunton, J; Leadlay, P F

    2001-09-01

    Using a previously developed expression system based on the erythromycin-producing strain of Saccharopolyspora erythraea, O-methyltransferases from the spinosyn biosynthetic gene cluster of Saccharopolyspora spinosa have been shown to modify a rhamnosyl sugar attached to a 14-membered polyketide macrolactone. The spnI, spnK and spnH methyltransferase genes were expressed individually in the S. erythraea mutant SGT2, which is blocked both in endogenous macrolide biosynthesis and in ery glycosyltransferases eryBV and eryCIII. Exogenous 3-O-rhamnosyl-erythronolide B was efficiently converted into 3-O-(2'-O-methylrhamnosyl)-erythronolide B by the S. erythraea SGT2 (spnI) strain only. When 3-O-(2'-O-methylrhamnosyl)-erythronolide B was, in turn, fed to a culture of S. erythraea SGT2 (spnK), 3-O-(2',3'-bis-O-methylrhamnosyl)-erythronolide B was identified in the culture supernatant, whereas S. erythraea SGT2 (spnH) was without effect. These results confirm the identity of the 2'- and 3'-O-methyltransferases, and the specific sequence in which they act, and they demonstrate that these methyltransferases may be used to methylate rhamnose units in other polyketide natural products with the same specificity as in the spinosyn pathway. In contrast, 3-O-(2',3'-bis-O-methylrhamnosyl)-erythronolide B was found not to be a substrate for the 4'-O-methyltransferase SpnH. Although rhamnosylerythromycins did not serve directly as substrates for the spinosyn methyltransferases, methylrhamnosyl-erythromycins were obtained by subsequent conversion of the corresponding methylrhamnosyl-erythronolide precursors using the S. erythraea strain SGT2 housing EryCIII, the desosaminyltransferase of the erythromycin pathway. 3-O-(2'-O-methylrhamnosyl)-erythromycin D was tested and found to be significantly active against a strain of erythromycin-sensitive Bacillus subtilis. PMID:11555300

  8. Yunnan-Guizhou Plateau in Summer and Autumn Broccoli Standardized Cultivation Techniques%云贵高原反季青花菜标准化栽培技术

    Institute of Scientific and Technical Information of China (English)

    刘兴平

    2012-01-01

    云贵高原夏秋季节气候温凉,适于种植反季青花菜,本文结合多年生产实践,从品种选择、培育壮苗、规格定植、肥水管理、病虫防治、适时采收等措施总结介绍了一套云贵高原冷凉山区反季青花菜标准化栽培技术。%Yunnan-Guizhou Plateau in summer and autumn climate was cool and suitable for growing-season broccoli, the paper summary description is a combination of years of production practices, species selection, nurturing seedlings, specifications planting, fertilizer and water management, pest control, timely harvest measuresYunnan-Guizhou Plateau in cold mountain-season broccoli standardization of cultivation techniques.

  9. The Drosophila G9a gene encodes a multi-catalytic histone methyltransferase required for normal development

    OpenAIRE

    Stabell, Marianne; Eskeland, Ragnhild; Bjørkmo, Mona; Larsson, Jan; Aalen, Reidunn B.; Imhof, Axel; Lambertsson, Andrew

    2006-01-01

    Mammalian G9a is a histone H3 Lys-9 (H3–K9) methyltransferase localized in euchromatin and acts as a co-regulator for specific transcription factors. G9a is required for proper development in mammals as g9a−/g9a− mice show growth retardation and early lethality. Here we describe the cloning, the biochemical and genetical analyses of the Drosophila homolog dG9a. We show that dG9a shares the structural organization of mammalian G9a, and that it is a multi-catalytic histone methyltransferase wit...

  10. Functional analysis of the N- and C-terminus of mammalian G9a histone H3 methyltransferase

    OpenAIRE

    Estève, Pierre-Olivier; Patnaik, Debasis; Chin, Hang Gyeong; Benner, Jack; Teitell, Michael A.; Pradhan, Sriharsa

    2005-01-01

    Methylation of lysine 9 (K9) in the N-terminus tail of histone H3 (H3) in chromatin is associated with transcriptionally silenced genes and is mediated by histone methyltransferases. Murine G9a is a 1263 amino acid H3-K9 methyltransferase that possesses characteristic SET domain and ANK repeats. In this paper, we have used a series of green fluorescent protein-tagged deletion constructs to identify two nuclear localization signals (NLS), the first NLS embedded between amino acids 24 and 109 a...

  11. CpG underrepresentation and the bacterial CpG-specific DNA methyltransferase M.MpeI

    OpenAIRE

    Wojciechowski, Marek; Czapinska, Honorata; Bochtler, Matthias

    2013-01-01

    Cytosine methylation promotes deamination. In eukaryotes, CpG methylation is thought to account for CpG underrepresentation. Whether scarcity of CpGs in prokaryotic genomes is diagnostic for methylation is not clear. Here, we report that Mycoplasms tend to be CpG depleted and to harbor a family of constitutively expressed or phase variable CpG-specific DNA methyltransferases. The very CpG poor Mycoplasma penetrans and its constitutively active CpG-specific methyltransferase M.MpeI were chosen...

  12. Active packaging for fresh-cut broccoli using 1-methylcyclopropene in biodegradable sachet/ Embalagem ativa para brócolis minimamente processado utilizando 1-metilciclopropeno em sachê biodegradável

    Directory of Open Access Journals (Sweden)

    Marta de Toledo Benassi

    2006-07-01

    Full Text Available Fresh-cut broccoli florets were packed in polypropylene pots containing a sachet with 1- methylcyclopropene (1-MCP and sealed with biodegradable starch-based film. Broccoli was stored for 8 days at 12oC and after this time the color and the texture of the product were similar of the fresh broccoli, with no off-flavor development or decay. Active packaging with 1-MCP in a sachet was efficient to extent shelf life of broccoli florets retarding yellowing and vitamin C losses. It is an alternative of 1-MCP treatment for fresh-cut products and besides, the sachet can absorbs condensed water reducing spoilage and off-odor development.Brócolis minimamente processados foram embalados em bandejas de polipropileno com sachê contendo 1-metilciclopropeno (1-MCP e selado com filme biodegradável de amido. Os brócolis foram armazenados por 8 dias a 12oC e após este tempo a cor e a textura mantiveram-se similares às do produto fresco, sem desenvolvimento de odor não característico ou podridão. A embalagem ativa contendo 1-MCP na forma de sachê foi eficiente no aumento da vida de prateleira de brócolis, retardando o amarelecimento e a perda de vitamina C. Esta é uma alternativa para o tratamento com 1-MCP para produtos minimamente processados e além disso, o sachê pode absorver a água condensada, reduzindo a deterioração e o desenvolvimento de odor não característico.

  13. UV-C对西兰花抗氧化活性及相关品质的影响%Changes of Antioxidant Activity, Compounds and Quality of Broccoli Florets During Different Temperatures

    Institute of Scientific and Technical Information of China (English)

    张怡; 关文强; 张娜; 刘莉莉; 阎瑞香; 孙德岭

    2014-01-01

    在0℃条件下,采用不同剂量UV-C(1、4、7 kJ/m2)照射处理西兰花后冷藏并测定相关品质指标。结果表明4 kJ/m2 UV-C照射处理增加了西兰花抗氧化活性,可以更好地延缓西兰花的衰老。在21天以后,4 kJ/m2 UV-C处理增高西兰花的叶绿素、类胡萝卜素和花青素的含量,延缓其降解。UV-C照射能很好地保持西兰花的感官品质,减慢黄化过程,其中4 kJ/m2 UV-C照射的综合感官效果最好。%At 0℃condition, the processing broccoli by different doses of UV-C (1, 4, 7 kJ/m2)irradiation were chilled and determined related quality indicators.The result showed that:4 kJ/m2 UV-C irradiation increased antioxidant activity of broccoli and resisted senility better. After 21 days, 4 kJ/m2 UV-C treatment can heighten the content of chlorophyll, carotenoids and anthocyanins of broccoli and resisted degradation. UV-C irradiation remain well organoleptic quality of broccoli, delayed the yellowing process. 4 kJ/m 2 UV-C irradiation of synthesis organoleptic effect is the best.

  14. Effect of Different Drying Methods on the Sulforaphane Content of Broccoli%不同干燥方法对西兰花中萝卜硫素含量影响

    Institute of Scientific and Technical Information of China (English)

    谢述琼; 何珺; 杨佳年; 陈震

    2013-01-01

    Drying Broccoli's flower,stalk and leaf with three usual drying methods which were drying in the shade,50 ℃-90 ℃drying and drying in the sun,indentifying sulforaphane content with HPLC. Sulforaphane content in broccoli's different parts was seed>flower>stalk>leaf,with different drying methods,drying was the best and drying in the shade was the worst,with the five drying temperature it was 90 ℃ that there were the highest sulforaphane content of broccoli. with the different parts the sulforaphane content of broccoli's seed was the highest,it had 10.48 mg/g;the best drying methods was 90℃drying.%  采用阴干、50℃~90℃烘干及晒干三种常用干燥方式干燥西兰花的花、茎、叶,采用HPLC测定萝卜硫素含量。西兰花不同部位中萝卜硫素含量是种子>花>茎>叶;而不同干燥方法中烘干最好,阴干最差,5种烘干温度中以90℃烘干时萝卜硫素含量最高。西兰花不同部位中种子萝卜硫素含量最高,有10.48 mg/g;而干燥方式采用90℃烘干最佳。

  15. 高温多雨条件下不同品种青花菜生育期变化规律研究%Change Rules of Growth Period of Different Broccoli Cultivars under High Temperature and Rainy Conditions

    Institute of Scientific and Technical Information of China (English)

    苏英京

    2012-01-01

      2011年是气候条件十分特殊的一年,在主要播种期播种的临海青花菜生育期严重提前,基地青花菜上市期严重偏离正常季节。研究发现,青花菜生育期变化存在环境敏感性,不同品种生育期变化敏感性不同,高温和多雨是其生育期变化的2个主导因素。%  The weather of 2011 was abnormal, which made the growth period and the time to the markets of broccoli advanced seriously. The results showed that the growth period of broccoli was sensitive to environment factors, and high temperature and rainy conditions were the two key factors for the changes of growth period of broccoli.

  16. 光照处理提高鲜切西兰花贮藏品质%Quality Improvement of Fresh-Cut Broccoli (Brassica Oleracea L.) during Storage by Light Illumination

    Institute of Scientific and Technical Information of China (English)

    詹丽娟; 魏国强; 乔明武; 王慧荣

    2012-01-01

    In this study, the effect of continuous light illumination at 2000 lx on the quality of fresh-cut broccoli during 7 days of cold storage was investigated by spectrophotometry. The results indicated that light illumination significantly alleviated chlorophyll degradation and yellowing occurrence during storage, and meanwhile could maintain carotenoids, total soluble solid and vitamin C contents as well as antioxidant capacity of fresh-cut broccoli during storage. However, light illumination also induced more weight loss of fresh-cut broccoli during the whole storage compared with dark storage.%为减缓鲜切西兰花贮藏期间品质下降,延长贮藏期,运用分光光度法研究连续光照处理(2000ix)对鲜切西兰花低温贮藏期间品质变化的影响。结果表明,和黑暗对照相比,光照处理显著减缓鲜切西兰花贮藏期间叶绿素降解和黄化发生率,维持类胡萝卜素、可溶性固形物和抗坏血酸含量,从而使其保持较高的抗氧化力。然而,光照处理显著促进鲜切西兰花鲜质量损失。

  17. 危害分析和关键环节控制点在西兰花冷链物流中的应用研究%The Application of HACCP in Cold-chain Logistics of Broccoli

    Institute of Scientific and Technical Information of China (English)

    张艳艳; 刘斌

    2013-01-01

    Cold-chain logistics protects harvest quality and reduce loss of broccoli in circulation field.In this paper,the application of HACCP system in cold-chain logistics of broccoli was studied.Hazard analysis was analyzed and four critical control points including pre-cool,package,cold storage and transportation were determined.The corresponding HACCP program was made,so as to keep original quality and improve shelf life of broccoli in circulation field.%全程冷链是减少西兰花流通环节的损耗、保持采收时质量的重要条件.将危害分析和关键环节控制点(HACCP)管理体系的预防性思维应用于西兰花的冷链物流,通过危害分析,确定了预冷、包装、冷藏、运输4个关键控制点并制定了HACCP计划表,以最大限度地保持西兰花的原有品质、延长货架期.

  18. Research of Controling Freezing Point of Broccoli by Using Freezing Point Regulators%冰点调节剂对西兰花冰点的控制研究

    Institute of Scientific and Technical Information of China (English)

    林本芳; 鲁晓翔; 李江阔

    2012-01-01

    本文研究不同冰点调节剂对西兰花冰点的影响.实验选取葡萄糖、维生素C、氯化钙和山梨醇四种冰点调节剂,利用单因素比较实验法,系统研究了冰点调节剂种类及其使用条件对西兰花冰点的影响.结果表明,氯化钙是较好的冰点调节剂;3%氯化钙浸泡30min对西兰花的冰点调节效果较好,使西兰花冰点从-0.8℃降到-1℃.%In this research, the effects of different freezing point regulators on broccoli freezing point were investigated. Four kinds of freezing point regulators, including glucose, vitamin C, calcium chloride and sorbitol, were used for research on the influence of freezing point of broccoli by the single factor comparison experiment method. The results indicated that calcium chloride and glucose were the better freezing point regulators. When broccoli was soaked for 30 minutes with calcium chloride content being of 3%, its' freezing point was decreased from -0.8℃to-1 ℃.

  19. Hippocampal dysfunction in the Euchromatin histone methyltransferase 1 heterozygous knockout mouse model for Kleefstra syndrome.

    Science.gov (United States)

    Balemans, Monique C M; Kasri, Nael Nadif; Kopanitsa, Maksym V; Afinowi, Nurudeen O; Ramakers, Ger; Peters, Theo A; Beynon, Andy J; Janssen, Sanne M; van Summeren, Rik C J; Eeftens, Jorine M; Eikelenboom, Nathalie; Benevento, Marco; Tachibana, Makoto; Shinkai, Yoichi; Kleefstra, Tjitske; van Bokhoven, Hans; Van der Zee, Catharina E E M

    2013-03-01

    Euchromatin histone methyltransferase 1 (EHMT1) is a highly conserved protein that catalyzes mono- and dimethylation of histone H3 lysine 9, thereby epigenetically regulating transcription. Kleefstra syndrome (KS), is caused by haploinsufficiency of the EHMT1 gene, and is an example of an emerging group of intellectual disability (ID) disorders caused by genes encoding epigenetic regulators of neuronal gene activity. Little is known about the mechanisms underlying this disorder, prompting us to study the Euchromatin histone methyltransferase 1 heterozygous knockout (Ehmt1(+/-)) mice as a model for KS. In agreement with the cognitive disturbances observed in patients with KS, we detected deficits in fear extinction learning and both novel and spatial object recognition in Ehmt1(+/-) mice. These learning and memory deficits were associated with a significant reduction in dendritic arborization and the number of mature spines in hippocampal CA1 pyramidal neurons of Ehmt1(+/-) mice. In-depth analysis of the electrophysiological properties of CA3-CA1 synapses revealed no differences in basal synaptic transmission or theta-burst induced long-term potentiation (LTP). However, paired-pulse facilitation (PPF) was significantly increased in Ehmt1(+/-) neurons, pointing to a potential deficiency in presynaptic neurotransmitter release. Accordingly, a reduction in the frequency of miniature excitatory post-synaptic currents (mEPSCs) was observed in Ehmt1(+/-) neurons. These data demonstrate that Ehmt1 haploinsufficiency in mice leads to learning deficits and synaptic dysfunction, providing a possible mechanism for the ID phenotype in patients with KS. PMID:23175442

  20. Protein arginine methyltransferase 5 (PRMT5) is a novel coactivator of constitutive androstane receptor (CAR).

    Science.gov (United States)

    Kanno, Yuichiro; Inajima, Jun; Kato, Sayaka; Matsumoto, Maika; Tokumoto, Chikako; Kure, Yuki; Inouye, Yoshio

    2015-03-27

    The constitutive androstane receptor (CAR) plays a key role in the expression of xenobiotic/steroid and drug metabolizing enzymes and their transporters. In this study, we demonstrated that protein arginine methyltransferase 5 (PRMT5) is a novel CAR-interacting protein. Furthermore, the PRMT-dependent induction of a CAR reporter gene, which was independent of methyltransferase activity, was enhanced in the presence of steroid receptor coactivator 1 (SRC1), peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1α) or DEAD box DNA/RNA helicase DP97. Using tetracycline inducible-hCAR system in HepG2 cells, we showed that knockdown of PRMT5 with small interfering RNA suppressed tetracycline -induced mRNA expression of CYP2B6 but not of CYP2C9 or CYP3A4. PRMT5 enhanced phenobarbital-mediated transactivation of a phenobarbital-responsive enhancer module (PBREM)-driven reporter gene in co-operation with PGC-1α in rat primary hepatocytes. Based on these findings, we suggest PRMT5 to be a gene (or promoter)-selective coactivator of CAR by mediating the formation of complexes between hCAR and appropriate coactivators. PMID:25721668

  1. PML-RARA requires DNA methyltransferase 3A to initiate acute promyelocytic leukemia.

    Science.gov (United States)

    Cole, Christopher B; Verdoni, Angela M; Ketkar, Shamika; Leight, Elizabeth R; Russler-Germain, David A; Lamprecht, Tamara L; Demeter, Ryan T; Magrini, Vincent; Ley, Timothy J

    2016-01-01

    The DNA methyltransferases DNMT3A and DNMT3B are primarily responsible for de novo methylation of specific cytosine residues in CpG dinucleotides during mammalian development. While loss-of-function mutations in DNMT3A are highly recurrent in acute myeloid leukemia (AML), DNMT3A mutations are almost never found in AML patients with translocations that create oncogenic fusion genes such as PML-RARA, RUNX1-RUNX1T1, and MLL-AF9. Here, we explored how DNMT3A is involved in the function of these fusion genes. We used retroviral vectors to express PML-RARA, RUNX1-RUNX1T1, or MLL-AF9 in bone marrow cells derived from WT or DNMT3A-deficient mice. Additionally, we examined the phenotypes of hematopoietic cells from Ctsg-PML-RARA mice, which express PML-RARA in early hematopoietic progenitors and myeloid precursors, with or without DNMT3A. We determined that the methyltransferase activity of DNMT3A, but not DNMT3B, is required for aberrant PML-RARA-driven self-renewal ex vivo and that DNMT3A is dispensable for RUNX1-RUNX1T1- and MLL-AF9-driven self-renewal. Furthermore, both the PML-RARA-driven competitive transplantation advantage and development of acute promyelocytic leukemia (APL) required DNMT3A. Together, these findings suggest that PML-RARA requires DNMT3A to initiate APL in mice. PMID:26595813

  2. Thiopurine methyltransferase activity in the erythrocytes of adults and children: and HPLC-linked assay.

    Science.gov (United States)

    Micheli, V; Jacomelli, G; Fioravanti, A; Morozzi, G; Marcolongo, R; Pompucci, G

    1997-03-18

    A non-radioactive method that uses reverse-phase high performance liquid chromatography is described for the determination of thiopurine methyltransferase (E.C. 2.1.1.67) activity in human erythrocytes. The method is based on the direct quantitation of 6-methyl-mercaptopurine produced from 6-mercaptopurine by crude erythrocyte lysates. The method is accurate and reliable and suitable for diagnostic use. Activity values in control adults ranged from 5 to 32 pmol/h/mg haemoglobin. The activity in the erythrocytes of adult males was significantly higher compared to females (21 +/- 5 and 15 +/- 8 pmol/h/mg haemoglobin, respectively). The activity measured in the erythrocytes of children (22 +/- 5 pmol/h/mg haemoglobin) did not show any significant difference compared to adults. Thiopurine methyltransferase activity was measured in a female patient with systemic sclerosis who developed severe bone marrow depression after treatment with azathioprine and allopurinol. Activity (6.3 +/- 0.5 pmol/h/mg haemoglobin) was found in the lowest range of controls thus supporting the hypothesis that it could be responsible for increased azathioprine cytotoxicity. PMID:9086303

  3. Crystallization and preliminary X-ray crystallographic studies of O-methyltransferase from Anabaena PCC 7120

    International Nuclear Information System (INIS)

    The O-methyltransferase (OMT) from the Anabaena PCC 7120 has been overexpressed in a soluble form in E. coli, purified and crystallized. The crystals belonged to space group C2221 and diffracted to 2.4 Å resolution. O-Methyltransferase (OMT) is a ubiquitous enzyme that exists in bacteria, plants and humans and catalyzes a methyl-transfer reaction using S-adenosyl-l-methionine as a methyl donor and a wide range of phenolics as acceptors. To investigate the structure and function of OMTs, omt from Anabaena PCC 7120 was cloned into expression vector pET21a and expressed in a soluble form in Escherichia coli strain BL21 (DE3). The recombinant OMT protein was purified to homogeneity using a two-step strategy. Crystals of OMT that diffracted to a resolution of 2.4 Å were obtained using the hanging-drop vapour-diffusion method. The crystals belonged to space group C2221, with unit-cell parameters a = 131.620, b = 227.994, c = 150.777 Å, α = β = γ = 90°. There are eight molecules per asymmetric unit

  4. Identification and expression profiling of DNA methyltransferases during development and stress conditions in Solanaceae.

    Science.gov (United States)

    Kumar, Rahul; Chauhan, Pankaj Kumar; Khurana, Ashima

    2016-09-01

    DNA methyltransferase (DMTase) enzymes contribute to plant development and stress responses by de novo establishment and subsequent maintenance of DNA methylation during replication. However, the molecular mechanism underlying this activity remains obscure, especially in crop species. Using DMTase homolog complement in six Solanaceae species, we demonstrated here that their number remained conserved in Solanum lineage, whereas it was expanded in both pepper and Nicotiana benthamiana. Non-synonymous vs synonymous (Ka/Ks) substitution ratio revealed that most of the Solanaceous DMTase homologs undergo purifying selection. The genomic sequences of tomato DMT homologs in its wild relative, Solanum pennellii, remained highly conserved in their exons and methyltransferase domains. Structure analysis further revealed highly similar folding of DMTase homologs and conservation in the residues participating in protein-protein interaction in Solanum lineage, whereas a considerable diversification was observed of pepper homologs. Transcript profiling of DMTases highlighted both similar and distinct expression patterns of tomato homologs in other species during fruit development and stress responses. Overall, our analysis provides a strong basis for in-depth exploration of both conserved as well as distinct functions of tomato DMTase homologs in other economically important Solanaceae species. PMID:27380018

  5. The Histone Methyltransferase Activity of MLL1 Is Dispensable for Hematopoiesis and Leukemogenesis

    Directory of Open Access Journals (Sweden)

    Bibhu P. Mishra

    2014-05-01

    Full Text Available Despite correlations between histone methyltransferase (HMT activity and gene regulation, direct evidence that HMT activity is responsible for gene activation is sparse. We address the role of the HMT activity for MLL1, a histone H3 lysine 4 (H3K4 methyltransferase critical for maintaining hematopoietic stem cells (HSCs. Here, we show that the SET domain, and thus HMT activity of MLL1, is dispensable for maintaining HSCs and supporting leukemogenesis driven by the MLL-AF9 fusion oncoprotein. Upon Mll1 deletion, histone H4 lysine 16 (H4K16 acetylation is selectively depleted at MLL1 target genes in conjunction with reduced transcription. Surprisingly, inhibition of SIRT1 is sufficient to prevent the loss of H4K16 acetylation and the reduction in MLL1 target gene expression. Thus, recruited MOF activity, and not the intrinsic HMT activity of MLL1, is central for the maintenance of HSC target genes. In addition, this work reveals a role for SIRT1 in opposing MLL1 function.

  6. Human catechol-O-methyltransferase: Cloning and expression of the membrane-associated form

    International Nuclear Information System (INIS)

    A cDNA clone for human catechol-O-methyltransferase was isolated from a human hepatoma cell line (Hep G2) cDNA library by hybridization screening with a porcine cDNA probe. The cDNA clone was sequenced and found to have an insert of 1226 nucleotides. The deduced primary structure of hCOMT is composed of 271 amino acid residues with the predicted molecular mass of 30 kDa. At its N terminus it has a hydrophobic segment of 21 amino acid residues that may be responsible for insertion of hCOMT into the endoplasmic reticulum membrane. The primary structure of hCOMT exhibits high homology to the porcine partial cDNA sequence (93%). The deduced amino acid sequence contains two tryptic peptide sequences (T-22, T-33) found in porcine liver catechol-O-methyltransferase (CEMT). The coding region of hCOMT cDNA was placed under the control of the cytomegalovirus promoter to transfect human kidney 293 cells. The recombinant hCOMT was shown by immunoblot analysis to be mainly associated with the membrane fraction. RNA blot analysis revealed one COMT mRNA transcript of 1.4 kilobases in Hep G2 poly(A)+ RNA

  7. Biochemical characterization of maintenance DNA methyltransferase DNMT-1 from silkworm, Bombyx mori.

    Science.gov (United States)

    Mitsudome, Takumi; Mon, Hiroaki; Xu, Jian; Li, Zhiqing; Lee, Jae Man; Patil, Anandrao Ashok; Masuda, Atsushi; Iiyama, Kazuhiro; Morokuma, Daisuke; Kusakabe, Takahiro

    2015-03-01

    DNA methylation is an important epigenetic mechanism involved in gene expression of vertebrates and invertebrates. In general, DNA methylation profile is established by de novo DNA methyltransferases (DNMT-3A, -3B) and maintainance DNA methyltransferase (DNMT-1). DNMT-1 has a strong substrate preference for hemimethylated DNA over the unmethylated one. Because the silkworm genome lacks an apparent homologue of de novo DNMT, it is still unclear that how silkworm chromosome establishes and maintains its DNA methylation profile. As the first step to unravel this enigma, we purified recombinant BmDNMT-1 using baculovirus expression system and characterized its DNA-binding and DNA methylation activity. We found that the BmDNMT-1 preferentially methylates hemimethylated DNA despite binding to both unmethylated and hemimethylated DNA. Interestingly, BmDNMT-1 formed a complex with DNA in the presence or absence of methyl group donor, S-Adenosylmethionine (AdoMet) and the AdoMet-dependent complex formation was facilitated by Zn(2+) and Mn(2+). Our results provide clear evidence that BmDNMT-1 retained the function as maintenance DNMT but its sensitivity to metal ions is different from mammalian DNMT-1. PMID:25623240

  8. Bacteriophage adenine methyltransferase: a life cycle regulator? Modelled using Vibrio harveyi myovirus like.

    Science.gov (United States)

    Bochow, S; Elliman, J; Owens, L

    2012-11-01

    The adenine methyltransferase (DAM) gene methylates GATC sequences that have been demonstrated in various bacteria to be a powerful gene regulator functioning as an epigenetic switch, particularly with virulence gene regulation. However, overproduction of DAM can lead to mutations, giving rise to variability that may be important for adaptation to environmental change. While most bacterial hosts carry a DAM gene, not all bacteriophage carry this gene. Currently, there is no literature regarding the role DAM plays in life cycle regulation of bacteriophage. Vibrio campbellii strain 642 carries the bacteriophage Vibrio harveyi myovirus like (VHML) that has been proven to increase virulence. The complete genome sequence of VHML bacteriophage revealed a putative adenine methyltransferase gene. Using VHML, a new model of phage life cycle regulation, where DAM plays a central role between the lysogenic and lytic states, will be hypothesized. In short, DAM methylates the rha antirepressor gene and once methylation is removed, homologous CI repressor protein becomes repressed and non-functional leading to the switching to the lytic cycle. Greater understanding of life cycle regulation at the genetic level can, in the future, lead to the genesis of chimeric bacteriophage with greater control over their life cycle for their safe use as probiotics within the aquaculture industry. PMID:22681538

  9. Improved radioenzymatic assay for plasma norepinephrine using purified phenylethanolamine n-methyltransferase

    International Nuclear Information System (INIS)

    Radioenzymatic assays have been developed for catecholamines using either catechol O-methyltransferase (COMT) or phenylethanolamine N-methyltransferase (PNMT). Assays using PNMT are specific for norepinephrine (NE) and require minimal manipulative effort but until now have been less sensitive than the more complex procedures using COMT. The authors report an improved purification scheme for bovine PNMT which has permitted development of an NE assay with dramatically improved sensitivity (0.5 pg), specificity and reproducibility (C.V. < 5%). PNMT was purified by sequential pH 5.0 treatment and dialysis and by column chromatographic procedures using DEAE-Sephacel, Sepharcryl S-200 and Phenyl-Boronate Agarose. Recovery of PNMT through the purification scheme was 50%, while blank recovery was <.001%. NE can be directly quantified in 25 ul of human plasma and an 80 tube assay can be completed within 4 h. The capillary to venous plasma NE gradient was examined in 8 normotensive male subjects. Capillary plasma (NE (211.2 +/- 61.3 pg/ml)) was lower than venous plasma NE (366.6 +/- 92.5 pg/ml) in all subjects (p < 0.005). This difference suggests that capillary (NE) may be a unique indicator of sympathetic nervous system activity in vivo. In conclusion, purification of PNMT has facilitated development of an improved radioenzymatic for NE with significantly improved sensitivity

  10. X-ray crystal structure of N-6 adenine deoxyribose nucleic acid methyltransferase from Streptococcus pneumoniae

    Science.gov (United States)

    Tran, Phidung Hong

    X-ray diffraction by using resonant anomalous scattering has become a popular tool for solving crystal structures in the last ten years with the expanded availability of tunable synchrotron radiation for protein crystallography. Mercury atoms were used for phasing. The crystal structure of N-6 deoxyribose nucleic acid methyltransferase from Streptoccocus pneumoniae (DpnM) was solved by using the Multiple Anomalous Diffraction technique. The crystal structure reveals the formation of mercaptide between the mercury ion and the thiol group on the cysteine amino acid in a hydrophobic environment. The crystal structure contains the bound ligand, S- adenosyl-l-methionine on the surface of the concave opening. The direction of the β-strands on the beta sheets are identical to other solved methyltransferases. The highly conserved motifs, DPPY and the FxGxG, are found to be important in ligand binding and possibly in methyl group transfer. The structure has a concave cleft with an opening on the order of 30 Å that can accommodate a DNA duplex. By molecular modelling coupled to sequence alignment, two other highly conserved residues Arg21 and Gly19 are found to be important in catalysis.

  11. Inactivating mutations of the histone methyltransferase gene EZH2 in myeloid disorders.

    Science.gov (United States)

    Ernst, Thomas; Chase, Andrew J; Score, Joannah; Hidalgo-Curtis, Claire E; Bryant, Catherine; Jones, Amy V; Waghorn, Katherine; Zoi, Katerina; Ross, Fiona M; Reiter, Andreas; Hochhaus, Andreas; Drexler, Hans G; Duncombe, Andrew; Cervantes, Francisco; Oscier, David; Boultwood, Jacqueline; Grand, Francis H; Cross, Nicholas C P

    2010-08-01

    Abnormalities of chromosome 7q are common in myeloid malignancies, but no specific target genes have yet been identified. Here, we describe the finding of homozygous EZH2 mutations in 9 of 12 individuals with 7q acquired uniparental disomy. Screening of a total of 614 individuals with myeloid disorders revealed 49 monoallelic or biallelic EZH2 mutations in 42 individuals; the mutations were found most commonly in those with myelodysplastic/myeloproliferative neoplasms (27 out of 219 individuals, or 12%) and in those with myelofibrosis (4 out of 30 individuals, or 13%). EZH2 encodes the catalytic subunit of the polycomb repressive complex 2 (PRC2), a highly conserved histone H3 lysine 27 (H3K27) methyltransferase that influences stem cell renewal by epigenetic repression of genes involved in cell fate decisions. EZH2 has oncogenic activity, and its overexpression has previously been causally linked to differentiation blocks in epithelial tumors. Notably, the mutations we identified resulted in premature chain termination or direct abrogation of histone methyltransferase activity, suggesting that EZH2 acts as a tumor suppressor for myeloid malignancies. PMID:20601953

  12. Conversion of nicotinic acid to trigonelline is catalyzed by N-methyltransferase belonged to motif B′ methyltransferase family in Coffea arabica

    Energy Technology Data Exchange (ETDEWEB)

    Mizuno, Kouichi, E-mail: koumno@akita-pu.ac.jp [Faculty of Bioresource Sciences, Akita Prefectural University, Akita City, Akita 010-0195 (Japan); Matsuzaki, Masahiro [Faculty of Bioresource Sciences, Akita Prefectural University, Akita City, Akita 010-0195 (Japan); Kanazawa, Shiho [Graduate School of Humanities and Sciences, Ochanomizu University, Otsuka, Bunkyo-ku, Tokyo 112-8610 (Japan); Tokiwano, Tetsuo; Yoshizawa, Yuko [Faculty of Bioresource Sciences, Akita Prefectural University, Akita City, Akita 010-0195 (Japan); Kato, Misako [Graduate School of Humanities and Sciences, Ochanomizu University, Otsuka, Bunkyo-ku, Tokyo 112-8610 (Japan)

    2014-10-03

    Graphical abstract: Trigonelline synthase catalyzes the conversion of nicotinic acid to trigonelline. We isolated and characterized trigonelline synthase gene(s) from Coffea arabica. - Highlights: • Trigonelline is a major compound in coffee been same as caffeine is. • We isolated and characterized trigonelline synthase gene. • Coffee trigonelline synthases are highly homologous with coffee caffeine synthases. • This study contributes the fully understanding of pyridine alkaloid metabolism. - Abstract: Trigonelline (N-methylnicotinate), a member of the pyridine alkaloids, accumulates in coffee beans along with caffeine. The biosynthetic pathway of trigonelline is not fully elucidated. While it is quite likely that the production of trigonelline from nicotinate is catalyzed by N-methyltransferase, as is caffeine synthase (CS), the enzyme(s) and gene(s) involved in N-methylation have not yet been characterized. It should be noted that, similar to caffeine, trigonelline accumulation is initiated during the development of coffee fruits. Interestingly, the expression profiles for two genes homologous to caffeine synthases were similar to the accumulation profile of trigonelline. We presumed that these two CS-homologous genes encoded trigonelline synthases. These genes were then expressed in Escherichiacoli, and the resulting recombinant enzymes that were obtained were characterized. Consequently, using the N-methyltransferase assay with S-adenosyl[methyl-{sup 14}C]methionine, it was confirmed that these recombinant enzymes catalyzed the conversion of nicotinate to trigonelline, coffee trigonelline synthases (termed CTgS1 and CTgS2) were highly identical (over 95% identity) to each other. The sequence homology between the CTgSs and coffee CCS1 was 82%. The pH-dependent activity curve of CTgS1 and CTgS2 revealed optimum activity at pH 7.5. Nicotinate was the specific methyl acceptor for CTgSs, and no activity was detected with any other nicotinate derivatives, or

  13. Conversion of nicotinic acid to trigonelline is catalyzed by N-methyltransferase belonged to motif B′ methyltransferase family in Coffea arabica

    International Nuclear Information System (INIS)

    Graphical abstract: Trigonelline synthase catalyzes the conversion of nicotinic acid to trigonelline. We isolated and characterized trigonelline synthase gene(s) from Coffea arabica. - Highlights: • Trigonelline is a major compound in coffee been same as caffeine is. • We isolated and characterized trigonelline synthase gene. • Coffee trigonelline synthases are highly homologous with coffee caffeine synthases. • This study contributes the fully understanding of pyridine alkaloid metabolism. - Abstract: Trigonelline (N-methylnicotinate), a member of the pyridine alkaloids, accumulates in coffee beans along with caffeine. The biosynthetic pathway of trigonelline is not fully elucidated. While it is quite likely that the production of trigonelline from nicotinate is catalyzed by N-methyltransferase, as is caffeine synthase (CS), the enzyme(s) and gene(s) involved in N-methylation have not yet been characterized. It should be noted that, similar to caffeine, trigonelline accumulation is initiated during the development of coffee fruits. Interestingly, the expression profiles for two genes homologous to caffeine synthases were similar to the accumulation profile of trigonelline. We presumed that these two CS-homologous genes encoded trigonelline synthases. These genes were then expressed in Escherichiacoli, and the resulting recombinant enzymes that were obtained were characterized. Consequently, using the N-methyltransferase assay with S-adenosyl[methyl-14C]methionine, it was confirmed that these recombinant enzymes catalyzed the conversion of nicotinate to trigonelline, coffee trigonelline synthases (termed CTgS1 and CTgS2) were highly identical (over 95% identity) to each other. The sequence homology between the CTgSs and coffee CCS1 was 82%. The pH-dependent activity curve of CTgS1 and CTgS2 revealed optimum activity at pH 7.5. Nicotinate was the specific methyl acceptor for CTgSs, and no activity was detected with any other nicotinate derivatives, or with

  14. Application of Tissue Culture, Molecular Marker and QTL Analysis in Broccoli Breeding%组织培养、分子标记和QT L技术在青花菜育种中的应用

    Institute of Scientific and Technical Information of China (English)

    张志仙; 朱长志; 檀国印; 王峰; 何道根

    2015-01-01

    育种是优异种质资源基因的重组。除传统新种质产生的方法外,通过组织培养、分子标记等技术来扩大物种的遗传基础已成为新种质资源形成的重要手段。青花菜,芸薹属甘蓝中以绿色花球为食用器官的一个变种,近年来在中国的种植面积越来越大,利用现代生物技术手段进行青花菜育种研究工作也越来越受到关注和重视。本文综述了游离小孢子培养、原生质体融合和转基因技术在青花菜种质资源创制中的应用,分析了各种方法存在的问题,并对青花菜遗传图谱构建、花球相关农艺性状及抗病性QTL/基因定位研究进行了探讨,并对其未来研究方向做了展望。%Breeding is kind of the genomic combination of excellent germplasms. Apart from conventional breeding methods generating new germplasms, approaches based on tissue culture and the molecular marker assisted breeding have become very popular biotechnologies to create the new breeding materials. Broccoli (Brassica oleracea L. var. italica Plenck), is an important vegetable worldwilde. In recent decades, broccoli production and consumption has risen significantly and the usage of biotechnologies on broccoli breeding has made great progress in China. This paper reviewed the uses of microspore culture, protoplast culture and fusion, transgenic technologies in improvement breeding of broccoli, and the limitation in those researches was mentioned as well. Moreover, construction of linkage maps, QTLs/genes for agronomically important broccoli traits including head morphologies and diseases resistance were analyzed in this review.

  15. Effect of Cold Acclimation Combined with Ice-Temperature Storage on the Quality of Broccolis%低温驯化结合冰温贮藏对西兰花品质的影响

    Institute of Scientific and Technical Information of China (English)

    林本芳; 鲁晓翔; 李江阔; 陈绍慧

    2012-01-01

    研究低温驯化结合冰温贮藏对西兰花保鲜品质的影响。贮藏期间每14d测定西兰花理化指标及营养成分。结果表明:与直接冰温贮藏或冷藏相比,低温驯化结合冰温贮藏处理对延缓西兰花VC含量和叶绿素含量的下降均有明显效果,且可降低呼吸强度和乙烯生成速率,提高西兰花过氧化物酶(POD)和过氧化氢酶(CAT)的活性,降低多酚氧化酶(PPO)活性,延缓相对电导率的升高。研究表明西兰花经低温驯化结合冰温贮藏的效果优于直接冰温贮藏的效果。%This study was conducted to investigate the effect of cold acclimation combined with ice-temperature storage(CACITS) on the quality of broccolis.Physicochemical indexes and nutritional components of broccolis were detected every 14 days during storage.The results show that the contents of VC and chlorophyll in broccoli were decreased more significantly by cold acclimation combined with ice-temperature storage than ice-temperature storage alone and ordinary cold storage;moreover,cold acclimation combined with ice-temperature storage could reduce respiratory intensity and ethylene generation rate in broccolis,increase peroxidase(POD) and catalase(CAT) activities,inhibit polyphenol oxidase(PPO) activity and delay the increase of relative electric conductivity.As a result,it can be concluded that cold acclimation combined with ice-temperature storage is better for the quality of of broccolis than ice-temperature storage alone.

  16. 不同出库方式对西兰花货架期品质的影响%Effect of different out-store tests on storage quality of broccolis during shelf-life period

    Institute of Scientific and Technical Information of China (English)

    林本芳; 鲁晓翔; 李江阔; 陈绍慧

    2012-01-01

    The effect of broccolis by different out-store tests after cold storage which temperature was -1 -1℃ was detected by analyzing physicochemical indexes and nutrition components. The research results showed that broccoli's Vc and chlorophyll content and firmness by low-temperature chain storage condition decreased less than that of progressively warming and control. Low-temperature chain storage increased superoxide dismutase (SOD) activity and slowed down polyphenol oxidase (PPO)activity, also decreased MDA content. Mildew spot index of broccolis was zero by low-temperature chain storage after 10d of out-store,whereas the values of progressively warming and control were 88.89% and 100% after 8d of out-store. Therefore,the effect of broccolis' shelf-life by different out-store tests was significant,the storage effect of broccolis by low- temperature chain storage was better than that of by progressively warming and control.%通过对西兰花在货架期间理化指标及营养成分变化规律的分析,探讨不同出库方式对冷藏(-1~1℃)后西兰花的保鲜效果。实验结果表明:与缓慢升温出库和直接出库(对照组)相比,低温冷链出库处理对延缓西兰花黄化以及Vc含量、叶绿素含量和硬度的下降均有明显效果,提高了西兰花超氧化物歧化酶(SOD)的活性,降低其多酚氧化酶(PPO)活性,延缓了丙二醛(MDA)含量的升高;该模式下货架期10d的西兰花仍没有生霉,而缓慢升温出库和直接出库的货架期8d时霉斑指数分别为88.89%和100%。这说明不同出库方式对西兰花的货架期有显著影响,低温冷链出库的西兰花保鲜效果更好。

  17. Burkholderia glumae ToxA Is a Dual-Specificity Methyltransferase That Catalyzes the Last Two Steps of Toxoflavin Biosynthesis.

    Science.gov (United States)

    Fenwick, Michael K; Philmus, Benjamin; Begley, Tadhg P; Ealick, Steven E

    2016-05-17

    Toxoflavin is a major virulence factor of the rice pathogen Burkholderia glumae. The tox operon of B. glumae contains five putative toxoflavin biosynthetic genes toxABCDE. ToxA is a predicted S-adenosylmethionine-dependent methyltransferase, and toxA knockouts of B. glumae are less virulent in plant infection models. In this study, we show that ToxA performs two consecutive methylations to convert the putative azapteridine intermediate, 1,6-didemethyltoxoflavin, to toxoflavin. In addition, we report a series of crystal structures of ToxA complexes that reveals the molecular basis of the dual methyltransferase activity. The results suggest sequential methylations with initial methylation at N6 of 1,6-didemethyltoxoflavin followed by methylation at N1. The two azapteridine orientations that position N6 or N1 for methylation are coplanar with a 140° rotation between them. The structure of ToxA contains a class I methyltransferase fold having an N-terminal extension that either closes over the active site or is largely disordered. The ordered conformation places Tyr7 at a position of a structurally conserved tyrosine site of unknown function in various methyltransferases. Crystal structures of ToxA-Y7F consistently show a closed active site, whereas structures of ToxA-Y7A consistently show an open active site, suggesting that the hydroxyl group of Tyr7 plays a role in opening and closing the active site during the multistep reaction. PMID:27070241

  18. Thirteen new patients with guanidinoacetate methyltransferase deficiency and functional characterization of nineteen novel missense variants in the GAMT gene

    DEFF Research Database (Denmark)

    Mercimek-Mahmutoglu, Saadet; Ndika, Joseph; Kanhai, Warsha;

    2014-01-01

    Guanidinoacetate methyltransferase deficiency (GAMT-D) is an autosomal recessively inherited disorder of creatine biosynthesis. Creatine deficiency on cranial proton magnetic resonance spectroscopy, and elevated guanidinoacetate levels in body fluids are the biomarkers of GAMT-D. In 74 patients 5...

  19. Impaired Homocysteine Transmethylation and Protein-Methyltransferase Activity Reduce Expression of Selenoprotein P: Implications for Obesity and Metabolic Syndrome

    Science.gov (United States)

    Obesity causes Metabolic Syndrome and Type-II Diabetes, disrupting hepatic function, methionine (Met)/homocysteine (Hcy) transmethylation and methyltransferase (PRMT) activities. Selenoprotein P (SEPP1), exported from the liver, is the predominate form of plasma selenium (Se) and the physiological S...

  20. The Cfr rRNA methyltransferase confers resistance to Phenicols, Lincosamides, Oxazolidinones, Pleuromutilins, and Streptogramin A antibiotics

    DEFF Research Database (Denmark)

    Long, K. S.; Poehlsgaard, Jacob; Kehrenberg, C.; Schwarz, S.; Vester, B.

    2006-01-01

    A novel multidrug resistance phenotype mediated by the Cfr rRNA methyltransferase is observed in Staphylococcus aureus and Escherichia coli. The cfr gene has previously been identified as a phenicol and lincosamide resistance gene on plasmids isolated from Staphylococcus spp. of animal origin and...

  1. No up-regulation of the phosphatidylethanolamine N-methyltransferase pathway and choline production by sex hormones in cats

    NARCIS (Netherlands)

    Valtolina, Chiara; Vaandrager, Arie B; Favier, Robert P; Robben, Joris H; Tuohetahuntila, Maidina; Kummeling, Anne; Jeusette, Isabelle; Rothuizen, Jan

    2015-01-01

    BACKGROUND: Feline hepatic lipidosis (FHL) is a common cholestatic disease affecting cats of any breed, age and sex. Both choline deficiency and low hepatic phosphatidylethanolamine N-methyltransferase (PEMT) activity are associated with hepatic lipidosis (HL) in humans, mice and rats. The PEMT expr

  2. Implications of fast-time scale dynamics of human DNA/RNA cytosine methyltransferases (DNMTs) for protein function

    Czech Academy of Sciences Publication Activity Database

    Evans, D. A.; Bronowska, Agnieszka Katarzyna

    2010-01-01

    Roč. 125, 3/6 (2010), s. 407-418. ISSN 1432-881X Institutional research plan: CEZ:AV0Z40550506 Keywords : MD simulations * DNA/RNA methyltransferase * enthalpy-entropy compensation Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.903, year: 2010

  3. Flexible double-headed cytosine-linked 2'-deoxycytidine nucleotides. Synthesis, polymerase incorporation to DNA and interaction with DNA methyltransferases

    Czech Academy of Sciences Publication Activity Database

    Kielkowski, Pavel; Cahová, Hana; Pohl, Radek; Hocek, Michal

    2016-01-01

    Roč. 24, č. 6 (2016), s. 1268-1276. ISSN 0968-0896 R&D Projects: GA ČR GBP206/12/G151 Institutional support: RVO:61388963 Keywords : nucleosides * nucleotides * pyrimidines * DNA methyltransferases * DNA polymerases Subject RIV: CC - Organic Chemistry Impact factor: 2.793, year: 2014

  4. An assay for human erythrocyte catechol-O-methyltransferase activity using a catechol estrogen as the substrate

    International Nuclear Information System (INIS)

    A radiometric assay for catechol-O-methyltransferase (COMT) activity in human erythrocytes is described that employs 2-hydroxy[3H]estrone, and nonradiolabeled S-adenosylmethionine (SAM) as the cosubstrates. The ease of separation of the product of the reaction, 2-methoxy[3H)estrone from 2-hydroxy-[3H]estrone makes it possible to achieve low reaction blanks. The assay is very sensitive, and only 200 μl of whole blood are used per determination. The assay is highly reproducible. The interassay variability (coefficient of variation) was 6.5% for 24 assays of COMT activity in red blood cells in blood obtained daily for 24 days from one person. In incubations conducted at 370C for 30 min, the catechol-O-methyltransferase activity was a linear function of enzyme concentration (equivalent to 11 to 180 μl of packed red blood cells). Employing this assay, we evaluated the catalytic conversion of 2-hydroxyestrone to 2-methoxy-estrone by catechol-O-methyltransferase from human red blood cells and found that the apparent Michaelis constant and the apparent maximal rate of reaction were 3 X 10-7 M and 6.7 X 10-9 mol.ml-1 erythrocytes . h-1, respectively. The catechol-O-methyltransferase activity measured in erythrocytes obtained from 100 healthy subjects (men and nonpregnant women) was 8.2 +- 0.17 (mean +- S.E.) nmol 2-methoxyestrone . ml-1 erythrocytes. h-1. (Auth.)

  5. Phylogenetic, molecular, and biochemical characterization of caffeic aicd O-methyltransferase (COMT) gene family in Brachypodium distachyon

    Science.gov (United States)

    Caffeic acid O-methyltransferase (COMT) is one of the important enzymes controlling lignin monomer production in plant cell wall synthesis. Analysis of the genome sequence of new grass model Brachypodium distachyon identified four COMT gene homologues, designated as BdCOMT1, BdCOMT2, BdCOMT3, and ...

  6. Molecular cloning, characterization and expression of the caffeic acid O-methyltransferase (COMT) ortholog from kenaf (Hibiscus cannabinus)

    Science.gov (United States)

    We cloned the full-length of the gene putatively encoding caffeic acid O-methyltransferase (COMT) from kenaf (Hibiscus cannabinus L.) using degenerate primers and the RACE (rapid amplification of cDNA ends) method. Kenaf is an herbaceous and rapidly growing dicotyledonous plant with great potential ...

  7. Current understanding of the interplay between catechol-O-methyltransferase genetic variants, sleep, brain development and cognitive performance in schizophrenia

    NARCIS (Netherlands)

    Tucci, Valter; Lassi, Glenda; Kas, Martien J

    2012-01-01

    Abnormal sleep is an endophenotype of schizophrenia. Here we provide an overview of the genetic mechanisms that link specific sleep physiological processes to schizophrenia-related cognitive defects. In particular, we will review the possible relationships between catechol-O-methyltransferase (COMT)

  8. Comparative Distribution and Retention of Arsenic in Arsenic (+3 Oxidation State) Methyltransferase Knockout and Wild Type Mice

    Science.gov (United States)

    The mouse arsenic (+3 oxidation state) methyltransferase (As3mt) gene encodes a ~ 43 kDa protein that catalyzes conversion of inorganic arsenic into methylated products. Heterologous expression of AS3MT or its silencing by RNA interference controls arsenic methylation phenotypes...

  9. Identification of 8-methyladenosine as the modification catalyzed by the radical SAM methyltransferase Cfr that confers antibiotic resistance in bacteria

    DEFF Research Database (Denmark)

    Giessing, Anders; Jensen, Søren Skov; Rasmussen, Anette;

    2009-01-01

    The Cfr methyltransferase confers combined resistance to five different classes of antibiotics that bind to the peptidyl transferase center of bacterial ribosomes. The Cfr-mediated modification has previously been shown to occur on nucleotide A2503 of 23S rRNA and has a mass corresponding to an a...

  10. Polymorphisms in O-methyltransferase genes are associated with stover cell wall digestibility in European maize (Zea mays L.)

    DEFF Research Database (Denmark)

    Brenner, Everton A; Zein, Imad; Chen, Yongsheng;

    2010-01-01

    Background OMT (O-methyltransferase) genes are involved in lignin biosynthesis, which relates to stover cell wall digestibility. Reduced lignin content is an important determinant of both forage quality and ethanol conversion efficiency of maize stover. Results Variation in genomic sequences codi...

  11. An integrated epigenetic and genetic analysis of DNA methyltransferase genes (DNMTs) in tumor resistant and susceptible chicken lines

    Science.gov (United States)

    Both epigenetic alterations and genetic variations play essential roles in tumorigenesis. The epigenetic modification of DNA methylation is catalyzed and maintained by the DNA methyltransferases (DNMT3a, DNMT3b and DNMT1). DNA mutations and DNA methylation profiles of DNMTs themselves and their rela...

  12. Methanol:coenzyme M methyltransferase from Methanosarcina barkeri. Zinc dependence and thermodynamics of the methanol:cob(I)alamin methyltransferase reaction.

    Science.gov (United States)

    Sauer, K; Thauer, R K

    1997-10-01

    In Methanosarcina barkeri, methanogenesis from methanol is initiated by the formation of methyl-coenzyme M from methanol and coenzyme M. This methyl transfer reaction is catalyzed by two enzymes, designated methyltransferases 1 (MT1) and 2 (MT2). Transferase MT1, which is composed of a 50-kDa subunit, MtaB, and a 27-kDa corrinoid-harbouring subunit, MtaC, has been shown recently to catalyze the methylation of free cob(I)alamin with methanol [Sauer, K., Harms, U. & Thauer, R. K. (1997) Eur. J. Biochem. 243, 670-677]. We report here that this reaction is catalyzed by subunit MtaB overproduced in Escherichia coli. MtaB also catalyzed the formation of methanol from methylcobalamin and H2O, the hydrolysis being associated with a free-energy change deltaG(o)' of approximately +7.0 kJ/mol. MtaB was found to contain 1 mol zinc, and its activity to be zinc dependent (pK(Zn2+) = 9.3). The zinc dependence of the MT2 (MtaA)-catalyzed reaction is also described (pK(Zn2+) = 9.6). PMID:9363780

  13. O6-methylguanine-DNA methyltransferase (MGMT): can function explain a suicidal mechanism?

    Science.gov (United States)

    Gouws, Chrisna; Pretorius, Pieter J

    2011-11-01

    Why does O(6)-methylguanine-DNA methyltransferase (MGMT), an indispensable DNA repair enzyme, have a mechanism which seems to run counter to its importance? This enzyme is key to the removal of detrimental alkyl adducts from guanine bases. Although the mechanism is well known, an unusual feature surrounds its mode of action, which is its so-called suicidal endpoint. In addition, induction of MGMT is highly variable and its kinetics is atypical. These features raise some questions on the seemingly paradoxical mechanism. In this manuscript we point out that, although there is ample literature regarding the "how" of the MGMT enzyme, we found a lack of information on "why" this specific mechanism is in place. We then ask whether we know all there is to know about MGMT, or if perhaps there is a further as yet unknown function for MGMT, or if the suicidal mechanism may play some kind of protective role in the cell. PMID:21864987

  14. Structural Basis for Binding of RNA and Cofactor by a KsgA Methyltransferase

    Energy Technology Data Exchange (ETDEWEB)

    Tu, Chao; Tropea, Joseph E.; Austin, Brian P.; Court, Donald L.; Waugh, David S.; Ji, Xinhua; (NCI)

    2009-03-27

    Among methyltransferases, KsgA and the reaction it catalyzes are conserved throughout evolution. However, the specifics of substrate recognition by the enzyme remain unknown. Here we report structures of Aquifex aeolicus KsgA, in its ligand-free form, in complex with RNA, and in complex with both RNA and S-adenosylhomocysteine (SAH, reaction product of cofactor S-adenosylmethionine), revealing critical structural information on KsgA-RNA and KsgA-SAH interactions. Moreover, the structures show how conformational changes that occur upon RNA binding create the cofactor-binding site. There are nine conserved functional motifs (motifs IVIII and X) in KsgA. Prior to RNA binding, motifs I and VIII are flexible, each exhibiting two distinct conformations. Upon RNA binding, the two motifs become stabilized in one of these conformations, which is compatible with the binding of SAH. Motif X, which is also stabilized upon RNA binding, is directly involved in the binding of SAH.

  15. Biochemical and Computational Analysis of the Substrate Specificities of Cfr and RlmN Methyltransferases

    DEFF Research Database (Denmark)

    Ntokou, Eleni; Hansen, Lykke Haastrup; Kongsted, Jacob;

    2015-01-01

    -ray structure of RlmN. We used a trinucleotide as target sequence and assessed its positioning at the active site for methylation. The calculations are in accordance with different poses of the trinucleotide in the two enzymes indicating major evolutionary changes to shift the C2/C8 specificities. To explore......Cfr and RlmN methyltransferases both modify adenine 2503 in 23S rRNA (Escherichia coli numbering). RlmN methylates position C2 of adenine while Cfr methylates position C8, and to a lesser extent C2, conferring antibiotic resistance to peptidyl transferase inhibitors. Cfr and RlmN show high sequence...... interchangeability between Cfr and RlmN we constructed various combinations of their genes. The function of the mixed genes was investigated by RNA primer extension analysis to reveal methylation at 23S rRNA position A2503 and by MIC analysis to reveal antibiotic resistance. The catalytic site is expected to be...

  16. Adding a Lysine Mimic in the Design of Potent Inhibitors of Histone Lysine Methyltransferases

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Yanqi; Ganesh, Thota; Horton, John R.; Spannhoff, Astrid; Liu, Jin; Sun, Aiming; Zhang, Xing; Bedford, Mark T.; Shinkai, Yoichi; Snyder, James P.; Cheng, Xiaodong (Emory); (Kyoto); (Texas)

    2010-07-19

    Dynamic histone lysine methylation involves the activities of modifying enzymes (writers), enzymes removing modifications (erasers), and readers of the histone code. One common feature of these activities is the recognition of lysines in methylated and unmethylated states, whether they are substrates, reaction products, or binding partners. We applied the concept of adding a lysine mimic to an established inhibitor (BIX-01294) of histone H3 lysine 9 methyltransferases G9a and G9a-like protein by including a 5-aminopentyloxy moiety, which is inserted into the target lysine-binding channel and becomes methylated by G9a-like protein, albeit slowly. The compound enhances its potency in vitro and reduces cell toxicity in vivo. We suggest that adding a lysine or methyl-lysine mimic should be considered in the design of small-molecule inhibitors for other methyl-lysine writers, erasers, and readers.

  17. Molecular basis for oncohistone H3 recognition by SETD2 methyltransferase.

    Science.gov (United States)

    Yang, Shuang; Zheng, Xiangdong; Lu, Chao; Li, Guo-Min; Allis, C David; Li, Haitao

    2016-07-15

    High-frequency point mutations of genes encoding histones have been identified recently as novel drivers in a number of tumors. Specifically, the H3K36M/I mutations were shown to be oncogenic in chondroblastomas and undifferentiated sarcomas by inhibiting H3K36 methyltransferases, including SETD2. Here we report the crystal structures of the SETD2 catalytic domain bound to H3K36M or H3K36I peptides with SAH (S-adenosylhomocysteine). In the complex structure, the catalytic domain adopts an open conformation, with the K36M/I peptide snuggly positioned in a newly formed substrate channel. Our structural and biochemical data reveal the molecular basis underying oncohistone recognition by and inhibition of SETD2. PMID:27474439

  18. Multi-site-specific 16S rRNA methyltransferase RsmF from Thermus thermophilus

    DEFF Research Database (Denmark)

    Demirci, Hasan; Larsen, Line H G; Hansen, Trine;

    2010-01-01

    Cells devote a significant effort toward the production of multiple modified nucleotides in rRNAs, which fine tune the ribosome function. Here, we report that two methyltransferases, RsmB and RsmF, are responsible for all four 5-methylcytidine (m(5)C) modifications in 16S rRNA of Thermus...... thermophilus. Like Escherichia coli RsmB, T. thermophilus RsmB produces m(5)C967. In contrast to E. coli RsmF, which introduces a single m(5)C1407 modification, T. thermophilus RsmF modifies three positions, generating m(5)C1400 and m(5)C1404 in addition to m(5)C1407. These three residues are clustered near...

  19. Nucleosome Binding Alters the Substrate Bonding Environment of Histone H3 Lysine 36 Methyltransferase NSD2.

    Science.gov (United States)

    Poulin, Myles B; Schneck, Jessica L; Matico, Rosalie E; Hou, Wangfang; McDevitt, Patrick J; Holbert, Marc; Schramm, Vern L

    2016-06-01

    Nuclear receptor-binding SET domain protein 2 (NSD2) is a histone H3 lysine 36 (H3K36)-specific methyltransferase enzyme that is overexpressed in a number of cancers, including multiple myeloma. NSD2 binds to S-adenosyl-l-methionine (SAM) and nucleosome substrates to catalyze the transfer of a methyl group from SAM to the ε-amino group of histone H3K36. Equilibrium binding isotope effects and density functional theory calculations indicate that the SAM methyl group is sterically constrained in complex with NSD2, and that this steric constraint is released upon nucleosome binding. Together, these results show that nucleosome binding to NSD2 induces a significant change in the chemical environment of enzyme-bound SAM. PMID:27183271

  20. Imatinib causes epigenetic alterations of PTEN gene via upregulation of DNA methyltransferases and polycomb group proteins

    International Nuclear Information System (INIS)

    We have recently reported the possible imatinib-resistant mechanism; long-term exposure of leukemia cells to imatinib downregulated levels of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) via hypermethylation of its promoter region (Leukemia 2010; 24: 1631). The present study explored the molecular mechanisms by which imatinib caused methylation on the promoter region of this tumor suppressor gene in leukemia cells. Real-time reverse transcription PCR found that long-term exposure of chronic eosinophilic leukemia EOL-1 cells expressing FIP1L1/platelet-derived growth factor receptor-α to imatinib induced expression of DNA methyltransferase 3A (DNMT3A) and histone-methyltransferase enhancer of zeste homolog 2 (EZH2), a family of polycomb group, thereby increasing methylation of the gene. Immunoprecipitation assay found the increased complex formation of DNMT3A and EZH2 proteins in these cells. Moreover, chromatin immunoprecipitation assay showed that amounts of both DNMT3A and EZH2 proteins bound around the promoter region of PTEN gene were increased in EOL-1 cells after exposure to imatinib. Furthermore, we found that levels of DNMT3A and EZH2 were strikingly increased in leukemia cells isolated from individuals with chronic myelogenous leukemia (n=1) and Philadelphia chromosome-positive acute lymphoblastic leukemia (n=2), who relapsed after treatment with imatinib compared with those isolated at their initial presentation. Taken together, imatinib could cause drug-resistance via recruitment of polycomb gene complex to the promoter region of the PTEN and downregulation of this gene's transcripts in leukemia patients

  1. Characterization of three O-methyltransferases involved in noscapine biosynthesis in opium poppy.

    Science.gov (United States)

    Dang, Thu-Thuy T; Facchini, Peter J

    2012-06-01

    Noscapine is a benzylisoquinoline alkaloid produced in opium poppy (Papaver somniferum) and other members of the Papaveraceae. It has been used as a cough suppressant and more recently was shown to possess anticancer activity. However, the biosynthesis of noscapine in opium poppy has not been established. A proposed pathway leading from (S)-reticuline to noscapine includes (S)-scoulerine, (S)-canadine, and (S)-N-methylcanadine as intermediates. Stem cDNA libraries and latex extracts of eight opium poppy cultivars displaying different alkaloid profiles were subjected to massively parallel pyrosequencing and liquid chromatography-tandem mass spectrometry, respectively. Comparative transcript and metabolite profiling revealed the occurrence of three cDNAs encoding O-methyltransferases designated as SOMT1, SOMT2, and SOMT3 that correlated with the accumulation of noscapine in the eight cultivars. SOMT transcripts were detected in all opium poppy organs but were most abundant in aerial organs, where noscapine primarily accumulates. SOMT2 and SOMT3 showed strict substrate specificity and regiospecificity as 9-O-methyltransferases targeting (S)-scoulerine. In contrast, SOMT1 was able to sequentially 9- and 2-O-methylate (S)-scoulerine, yielding (S)-tetrahydropalmatine. SOMT1 also sequentially 3'- and 7-O-methylated both (S)-norreticuline and (S)-reticuline with relatively high substrate affinity, yielding (S)-tetrahydropapaverine and (S)-laudanosine, respectively. The metabolic functions of SOMT1, SOMT2, and SOMT3 were investigated in planta using virus-induced gene silencing. Reduction of SOMT1 or SOMT2 transcript levels resulted in a significant decrease in noscapine accumulation. Reduced SOMT1 transcript levels also caused a decrease in papaverine accumulation, confirming the selective roles for these enzymes in the biosynthesis of both alkaloids in opium poppy. PMID:22535422

  2. Protein arginine methyltransferase 5 (PRMT5) is a novel coactivator of constitutive androstane receptor (CAR)

    International Nuclear Information System (INIS)

    The constitutive androstane receptor (CAR) plays a key role in the expression of xenobiotic/steroid and drug metabolizing enzymes and their transporters. In this study, we demonstrated that protein arginine methyltransferase 5 (PRMT5) is a novel CAR-interacting protein. Furthermore, the PRMT-dependent induction of a CAR reporter gene, which was independent of methyltransferase activity, was enhanced in the presence of steroid receptor coactivator 1 (SRC1), peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1α) or DEAD box DNA/RNA helicase DP97. Using tetracycline inducible-hCAR system in HepG2 cells, we showed that knockdown of PRMT5 with small interfering RNA suppressed tetracycline -induced mRNA expression of CYP2B6 but not of CYP2C9 or CYP3A4. PRMT5 enhanced phenobarbital-mediated transactivation of a phenobarbital-responsive enhancer module (PBREM)-driven reporter gene in co-operation with PGC-1α in rat primary hepatocytes. Based on these findings, we suggest PRMT5 to be a gene (or promoter)-selective coactivator of CAR by mediating the formation of complexes between hCAR and appropriate coactivators. - Highlights: • Nuclear receptor CAR interact with PRMT5. • PRMT5 enhances transcriptional activity of CAR. • PRMT5 synergistically enhances transactivity of CAR by the co-expression of SRC-1, DP97 or PGC1α. • PRMT5 is a gene-selective co-activator for hCAR

  3. Protein arginine methyltransferase 5 (PRMT5) is a novel coactivator of constitutive androstane receptor (CAR)

    Energy Technology Data Exchange (ETDEWEB)

    Kanno, Yuichiro, E-mail: ykanno@phar.toho-u.ac.jp; Inajima, Jun; Kato, Sayaka; Matsumoto, Maika; Tokumoto, Chikako; Kure, Yuki; Inouye, Yoshio

    2015-03-27

    The constitutive androstane receptor (CAR) plays a key role in the expression of xenobiotic/steroid and drug metabolizing enzymes and their transporters. In this study, we demonstrated that protein arginine methyltransferase 5 (PRMT5) is a novel CAR-interacting protein. Furthermore, the PRMT-dependent induction of a CAR reporter gene, which was independent of methyltransferase activity, was enhanced in the presence of steroid receptor coactivator 1 (SRC1), peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1α) or DEAD box DNA/RNA helicase DP97. Using tetracycline inducible-hCAR system in HepG2 cells, we showed that knockdown of PRMT5 with small interfering RNA suppressed tetracycline -induced mRNA expression of CYP2B6 but not of CYP2C9 or CYP3A4. PRMT5 enhanced phenobarbital-mediated transactivation of a phenobarbital-responsive enhancer module (PBREM)-driven reporter gene in co-operation with PGC-1α in rat primary hepatocytes. Based on these findings, we suggest PRMT5 to be a gene (or promoter)-selective coactivator of CAR by mediating the formation of complexes between hCAR and appropriate coactivators. - Highlights: • Nuclear receptor CAR interact with PRMT5. • PRMT5 enhances transcriptional activity of CAR. • PRMT5 synergistically enhances transactivity of CAR by the co-expression of SRC-1, DP97 or PGC1α. • PRMT5 is a gene-selective co-activator for hCAR.

  4. Functional characterisation of three o-methyltransferases involved in the biosynthesis of phenolglycolipids in Mycobacterium tuberculosis.

    Directory of Open Access Journals (Sweden)

    Roxane Simeone

    Full Text Available Phenolic glycolipids are produced by a very limited number of slow-growing mycobacterial species, most of which are pathogen for humans. In Mycobacterium tuberculosis, the etiologic agent of tuberculosis, these molecules play a role in the pathogenicity by modulating the host immune response during infection. The major variant of phenolic glycolipids produced by M. tuberculosis, named PGL-tb, consists of a large lipid core terminated by a glycosylated aromatic nucleus. The carbohydrate part is composed of three sugar residues, two rhamnosyl units and a terminal fucosyl residue, which is per-O-methylated, and seems to be important for pathogenicity. While most of the genes responsible for the synthesis of the lipid core domain and the saccharide appendage of PGL-tb have been characterized, the enzymes involved in the O-methylation of the fucosyl residue of PGL-tb remain unknown. In this study we report the identification and characterization of the methyltransferases required for the O-methylation of the terminal fucosyl residue of PGL-tb. These enzymes are encoded by genes Rv2954c, Rv2955c and Rv2956. Mutants of M. tuberculosis harboring deletion within these genes were constructed. Purification and analysis of the phenolglycolipids produced by these strains, using a combination of mass spectrometry and NMR spectroscopy, revealed that Rv2954c, Rv2955c and Rv2956 encode the methyltransferases that respectively catalysed the O-methylation of the hydroxyl groups located at positions 3, 4 and 2 of the terminal fucosyl residue of PGL-tb. Our data also suggest that methylation at these positions is a sequential process, starting with position 2, followed by positions 4 and 3.

  5. Expression of a functional jasmonic acid carboxyl methyltransferase is negatively correlated with strawberry fruit development.

    Science.gov (United States)

    Preuß, Anja; Augustin, Christiane; Figueroa, Carlos R; Hoffmann, Thomas; Valpuesta, Victoriano; Sevilla, José F; Schwab, Wilfried

    2014-09-15

    The volatile metabolite methyl jasmonate (MeJA) plays an important role in intra- and interplant communication and is involved in diverse biological processes. In this study, we report the cloning and functional characterization of a S-adenosyl-l-methionine:jasmonic acid carboxyl methyltransferase (JMT) from Fragaria vesca and Fragaria×ananassa. Biochemical assays and comprehensive transcript analyses showed that JMT has been erroneously annotated as gene fusion with a carboxyl methyltransferase (CMT) (gene15184) in the first published genome sequence of F. vesca. Recombinant FvJMT catalyzed the formation of MeJA with KM value of 22.3μM while FvCMT and the fusion protein were almost inactive. Activity of JMT with benzoic acid and salicylic acid as substrates was less than 1.5% of that with JA. Leucine at position 245, an amino acid missing in other JMT sequences is essential for activity of FvJMT. In accordance with MeJA levels, JMT transcript levels decreased steadily during strawberry fruit ripening, as did the expression levels of JA biosynthesis and regulatory genes. It appears that CMT has originated by a recent duplication of JMT and lost its enzymatic activity toward JA. In the newest version of the strawberry genome sequence (June 2014) CMT and JMT are annotated as separate genes in accordance with differential temporal and spatial expression patterns of both genes in Fragaria sp. In conclusion, MeJA, the inactive derivative of JA, is probably involved in early steps of fruit development by modulating the levels of the active plant hormone JA. PMID:25046752

  6. Computational Investigation of the Interplay of Substrate Positioning and Reactivity in Catechol O-Methyltransferase.

    Science.gov (United States)

    Patra, Niladri; Ioannidis, Efthymios I; Kulik, Heather J

    2016-01-01

    Catechol O-methyltransferase (COMT) is a SAM- and Mg2+-dependent methyltransferase that regulates neurotransmitters through methylation. Simulations and experiments have identified divergent catecholamine substrate orientations in the COMT active site: molecular dynamics simulations have favored a monodentate coordination of catecholate substrates to the active site Mg2+, and crystal structures instead preserve bidentate coordination along with short (2.65 Å) methyl donor-acceptor distances. We carry out longer dynamics (up to 350 ns) to quantify interconversion between bidentate and monodentate binding poses. We provide a systematic determination of the relative free energy of the monodentate and bidentate structures in order to identify whether structural differences alter the nature of the methyl transfer mechanism and source of enzymatic rate enhancement. We demonstrate that the bidentate and monodentate binding modes are close in energy but separated by a 7 kcal/mol free energy barrier. Analysis of interactions in the two binding modes reveals that the driving force for monodentate catecholate orientations in classical molecular dynamics simulations is derived from stronger electrostatic stabilization afforded by alternate Mg2+ coordination with strongly charged active site carboxylates. Mixed semi-empirical-classical (SQM/MM) substrate C-O distances (2.7 Å) for the bidentate case are in excellent agreement with COMT X-ray crystal structures, as long as charge transfer between the substrates, Mg2+, and surrounding ligands is permitted. SQM/MM free energy barriers for methyl transfer from bidentate and monodentate catecholate configurations are comparable at around 21-22 kcal/mol, in good agreement with experiment (18-19 kcal/mol). Overall, the work suggests that both binding poses are viable for methyl transfer, and accurate descriptions of charge transfer and electrostatics are needed to provide balanced relative barriers when multiple binding poses are

  7. Molecular and biochemical characterization of the jasmonic acid methyltransferase gene from black cottonwood (Populus trichocarpa)

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Nan [ORNL; Yao, Jianzhuang [University of Tennessee, Knoxville (UTK); Chaiprasongsuk, Minta [University of Tennessee, Knoxville (UTK); Li, Guanglin [University of Tennessee, Knoxville (UTK); Guan, Ju [University of Tennessee, Knoxville (UTK); Tschaplinski, Timothy J [ORNL; Guo, Hong [University of Tennessee, Knoxville (UTK); Chen, Feng [University of Tennessee, Knoxville (UTK)

    2013-01-01

    Methyl jasmonate is a metabolite known to be produced by many plants and has roles in diverse biological processes. It is biosynthesized by the action of S-adenosyl-L-methionine:jasmonic acid carboxyl methyltransferase (JMT), which belongs to the SABATH family of methyltransferases. Herein is reported the isolation and biochemical characterization of a JMT gene from black cottonwood (Populus trichocarpa). The genome of P. trichocarpa contains 28 SABATH genes (PtSABATH1 to PtSABATH28). Recombinant PtSABATH3 expressed in Escherichia coli showed the highest level of activity with jasmonic acid (JA) among carboxylic acids tested. It was therefore renamed PtJMT1. PtJMT1 also displayed activity with benzoic acid (BA), with which the activity was about 22% of that with JA. PtSABATH2 and PtSABATH4 were most similar to PtJMT1 among all PtSABATHs. However, neither of them had activity with JA. The apparent Km values of PtJMT1 using JA and BA as substrate were 175 lM and 341 lM, respectively. Mutation of Ser-153 and Asn-361, two residues in the active site of PtJMT1, to Tyr and Ser respectively, led to higher specific activity with BA than with JA. Homology-based structural modeling indicated that substrate alignment, in which Asn-361 is involved, plays a role in determining the substrate specificity of PtJMT1. In the leaves of young seedlings of black cottonwood, the expression of PtJMT1 was induced by plant defense signal molecules methyl jasmonate and salicylic acid and a fungal elicitor alamethicin, suggesting that PtJMT1 may have a role in plant defense against biotic stresses. Phylogenetic analysis suggests that PtJMT1 shares a common ancestor with the Arabidopsis JMT, and functional divergence of these two apparent JMT orthologs has occurred since the split of poplar and Arabidopsis lineages.

  8. Binding induced RNA conformational changes control substrate recognition and catalysis by the thiostrepton resistance methyltransferase (Tsr).

    Science.gov (United States)

    Kuiper, Emily G; Conn, Graeme L

    2014-09-19

    Ribosomal RNA (rRNA) post-transcriptional modifications are essential for ribosome maturation, translational fidelity, and are one mechanism used by both antibiotic-producing and pathogenic bacteria to resist the effects of antibiotics that target the ribosome. The thiostrepton producer Streptomyces azureus prevents self-intoxication by expressing the thiostrepton-resistance methyltransferase (Tsr), which methylates the 2'-hydroxyl of 23 S rRNA nucleotide adenosine 1067 within the thiostrepton binding site. Tsr is a homodimer with each protomer containing an L30e-like amino-terminal domain (NTD) and a SPOUT methyltransferase family catalytic carboxyl-terminal domain (CTD). We show that both enzyme domains are required for high affinity RNA substrate binding. The Tsr-CTD has intrinsic, weak RNA affinity that is necessary to direct the specific high-affinity Tsr-RNA interaction via NTDs, which have no detectable RNA affinity in isolation. RNA structure probing experiments identify the Tsr footprint on the RNA and structural changes in the substrate, induced specifically upon NTD binding, which are necessary for catalysis by the CTD. Additionally, we identify a key amino acid in each domain responsible for CTD-RNA binding and the observed NTD-dependent RNA structural changes. These studies allow us to develop a model for Tsr-RNA interaction in which the coordinated substrate recognition of each Tsr structural domain is an obligatory pre-catalytic recognition event. Our findings underscore the complexity of substrate recognition by RNA modification enzymes and the potential for direct involvement of the RNA substrate in controlling the process of its modification. PMID:25086036

  9. Induction of phase 2 antioxidant enzymes by broccoli sulforaphane: Perspectives in maintaining the antioxidant activity of vitamins A, C, and E

    Directory of Open Access Journals (Sweden)

    Sekhar s Boddupalli

    2012-01-01

    Full Text Available Consumption of fruits and vegetables is recognized as an important part of a healthy diet. Increased consumption of cruciferous vegetables in particular has been associated with a decreased risk of several degenerative and chronic diseases, including cardiovascular disease and certain cancers. Members of the cruciferous vegetable family, which includes broccoli, Brussels sprouts, cauliflower, and cabbage, accumulate significant concentrations of glucosinolates, which are metabolized in vivo to biologically active isothiocyanates (ITCs. The ITC sulforaphane, which is derived from glucoraphanin, has garnered particular interest due to its extraordinary ability to induce expression of several enzymes via the Keap1/Nrf2/ARE pathway. Nrf2/ARE gene products are typically characterized as Phase II detoxification enzymes and/or antioxidant (AO enzymes. Over the last decade, human clinical studies have begun to provide in vivo evidence of both Phase II and AO enzyme induction by SF. Many AO enzymes are redox cycling enzymes that maintain redox homeostasis and activity of free radical scavengers such as such as vitamins A, C and E. In this review, we present the existing evidence for induction of PII and AO enzymes by SF, the interactions of SF-induced AO enzymes and proposed maintenance of the essential vitamins A, C and E, and, finally, the current view of genotypic effects on ITC metabolism and AO enzyme induction and function.

  10. Ultrasonic-assisted enzymatic extraction of phenolics from broccoli (Brassica oleracea L. var. italica) inflorescences and evaluation of antioxidant activity in vitro.

    Science.gov (United States)

    Wu, Hao; Zhu, Junxiang; Yang, Long; Wang, Ran; Wang, Chengrong

    2015-06-01

    An efficient ultrasonic-assisted enzymatic extraction technique was applied to extracting phenolics from broccoli inflorescences without organic solvents. The synergistic model of enzymolysis and ultrasonication simultaneously was selected, and the enzyme combination was optimized by orthogonal test: cellulase 7.5 mg/g FW (fresh weight), pectinase 10 mg/g FW, and papain 1.0 mg/g FW. The operating parameters in ultrasonic-assisted enzymatic extraction were optimized with response surface methodology using Box-Behnken design. The optimal extraction conditions were as follows: ultrasonic power, 440 W; liquid to material ratio, 7.0:1 mL/g; pH value of 6.0 at 54.5 ℃ for 10 min. Under these conditions, the extraction yield of phenolics achieved 1.816 ± 0.0187 mg gallic acid equivalents/gram FW. The free radical scavenging activity of ultrasonic-assisted enzymatic extraction extracts was determined by 1,1-diphenyl-2-picrylhydrazyl·assay with EC50 values of 0.25, and total antioxidant activity was determined by ferric reducing antioxidant power assay with ferric reducing antioxidant power value of 0.998 mmol FeSO4/g compared with the referential ascorbic acid of 1.184 mmol FeSO4/g. PMID:24837595

  11. Glucoraphanin, the bioprecursor of the widely extolled chemopreventive agent sulforaphane found in broccoli, induces Phase-I xenobiotic metabolizing enzymes and increases free radical generation in rat liver

    International Nuclear Information System (INIS)

    Epidemiological and animal studies linking high fruit and vegetable consumption to lower cancer risk have strengthened the belief that long-term administration of isolated naturally occurring dietary constituents could reduce the risk of cancer. In recent years, metabolites derived from phytoalexins, such as glucoraphanin found in broccoli and other cruciferous vegetables (Brassicaceae), have gained much attention as potential cancer chemopreventive agents. The protective effect of these micronutrients is assumed to be due to the inhibition of Phase-I carcinogen-bioactivating enzymes and/or induction of Phase-II detoxifying enzymes, an assumption that still remains uncertain. The protective effect of glucoraphanin is thought to be due to sulforaphane, an isothiocyanate metabolite produced from glucoraphanin by myrosinase. Here we show, in rat liver, that while glucoraphanin slightly induces Phase-II enzymes, it powerfully boosts Phase-I enzymes, including activators of polycyclic aromatic hydrocarbons (PAHs), nitrosamines and olefins. Induction of the cytochrome P450 (CYP) isoforms CYP1A1/2, CYP3A1/2 and CYP2E1 was confirmed by Western immunoblotting. CYP induction was paralleled by an increase in the corresponding mRNA levels. Concomitant with this Phase-I induction, we also found that glucoraphanin generated large amount of various reactive radical species, as determined by electron paramagnetic resonance (EPR) spectrometry coupled to a radical-probe technique. This suggests that long-term uncontrolled administration of glucoraphanin could actually pose a potential health hazard

  12. Glucoraphanin, the bioprecursor of the widely extolled chemopreventive agent sulforaphane found in broccoli, induces Phase-I xenobiotic metabolizing enzymes and increases free radical generation in rat liver

    Energy Technology Data Exchange (ETDEWEB)

    Perocco, Paolo [Department of Experimental Pathology, Cancerology Section, viale Filopanti 22, I-40126, University of Bologna, Bologna (Italy); Bronzetti, Giorgio [Institute of Biology and Agricultural Biotechnology - CNR Research Area, via Moruzzi, I-56124 Pisa (Italy); Canistro, Donatella; Sapone, Andrea; Affatato, Alessandra; Pozzetti, Laura; Broccoli, Massimiliano [Department of Pharmacology, Molecular Toxicology Unit, via Irnerio 48, I-40126, University of Bologna, Bologna (Italy); Valgimigli, Luca [Department of Organic Chemistry ' A. Mangini' , Viale Risorgimento 4, I-40127, Alma-Mater Studiorum, University of Bologna, Bologna (Italy); Pedulli, Gian Franco [Department of Organic Chemistry ' A. Mangini' , Viale Risorgimento 4, I-40127, Alma-Mater Studiorum, University of Bologna, Bologna (Italy); Iori, Renato [C.R.A - Research Institute for Industrial Crops, via di Corticella 133, I-40129 Bologna (Italy); Barillari, Jessica [Institute of Biology and Agricultural Biotechnology - CNR Research Area, via Moruzzi, I-56124 Pisa (Italy)]|[C.R.A - Research Institute for Industrial Crops, via di Corticella 133, I-40129 Bologna (Italy); Sblendorio, Valeriana [Department of Pharmacology, Molecular Toxicology Unit, via Irnerio 48, I-40126, University of Bologna, Bologna (Italy); Legator, Marvin S. [Department of Preventive Medicine and Community Health, Division of Environmental Toxicology, The University of Texas Medical Branch at Galveston, 700 Harborside Drive, Galveston, TX 77555-1110 (United States); Paolini, Moreno [Department of Pharmacology, Molecular Toxicology Unit, via Irnerio 48, I-40126, University of Bologna, Bologna (Italy); Abdel-Rahman, Sherif Z. [Department of Preventive Medicine and Community Health, Division of Environmental Toxicology, The University of Texas Medical Branch at Galveston, 700 Harborside Drive, Galveston, TX 77555-1110 (United States)]. E-mail: sabdelra@utmb.edu

    2006-03-20

    Epidemiological and animal studies linking high fruit and vegetable consumption to lower cancer risk have strengthened the belief that long-term administration of isolated naturally occurring dietary constituents could reduce the risk of cancer. In recent years, metabolites derived from phytoalexins, such as glucoraphanin found in broccoli and other cruciferous vegetables (Brassicaceae), have gained much attention as potential cancer chemopreventive agents. The protective effect of these micronutrients is assumed to be due to the inhibition of Phase-I carcinogen-bioactivating enzymes and/or induction of Phase-II detoxifying enzymes, an assumption that still remains uncertain. The protective effect of glucoraphanin is thought to be due to sulforaphane, an isothiocyanate metabolite produced from glucoraphanin by myrosinase. Here we show, in rat liver, that while glucoraphanin slightly induces Phase-II enzymes, it powerfully boosts Phase-I enzymes, including activators of polycyclic aromatic hydrocarbons (PAHs), nitrosamines and olefins. Induction of the cytochrome P450 (CYP) isoforms CYP1A1/2, CYP3A1/2 and CYP2E1 was confirmed by Western immunoblotting. CYP induction was paralleled by an increase in the corresponding mRNA levels. Concomitant with this Phase-I induction, we also found that glucoraphanin generated large amount of various reactive radical species, as determined by electron paramagnetic resonance (EPR) spectrometry coupled to a radical-probe technique. This suggests that long-term uncontrolled administration of glucoraphanin could actually pose a potential health hazard.

  13. Large-Scale, Protection-Free Synthesis of Se-Adenosyl-l-selenomethionine Analogues and Their Application as Cofactor Surrogates of Methyltransferases

    OpenAIRE

    Bothwell, Ian R.; Luo, Minkui

    2014-01-01

    S-Adenosyl-l-methionine (SAM) analogues have previously demonstrated their utility as chemical reporters of methyltransferases. Here we describe the facile, large-scale synthesis of Se-alkyl Se-adenosyl-l-selenomethionine (SeAM) analogues and their precursor, Se-adenosyl-l-selenohomocysteine (SeAH). Comparison of SeAM analogues with their equivalent SAM analogues suggests that sulfonium-to-selenonium substitution can enhance their compatibility with certain protein methyltransferases, favorin...

  14. Interplay among coactivator-associated arginine methyltransferase 1, CBP, and CIITA in IFN-γ-inducible MHC-II gene expression

    OpenAIRE

    Zika, Eleni; Fauquier, Lucas; Vandel, Laurence; Ting, Jenny P -Y

    2005-01-01

    Class II major histocompatibility (MHC-II) genes are prototype targets of IFN-γ. IFN-γ activates the expression of the non-DNA-binding master regulator of MHC-II, class II transactivator (CIITA), which is crucial for enhanceosome formation and gene activation. This report shows the importance of the histone methyltransferase, coactivator-associated arginine methyltransferase (CARM1/PRMT4), during IFN-γ-induced MHC-II gene activation. It also demonstrates the coordinated regulation of CIITA, C...

  15. O6-Methylguanine-DNA Methyltransferase (MGMT) mRNA Expression Predicts Outcome in Malignant Glioma Independent of MGMT Promoter Methylation

    OpenAIRE

    Simone Kreth; Niklas Thon; Sabina Eigenbrod; Juergen Lutz; Carola Ledderose; Rupert Egensperger; Tonn, Joerg C.; Kretzschmar, Hans A.; Ludwig C Hinske; Kreth, Friedrich W.

    2011-01-01

    Background: We analyzed prospectively whether MGMT (O(6)-methylguanine-DNA methyltransferase) mRNA expression gains prognostic/predictive impact independent of MGMT promoter methylation in malignant glioma patients undergoing radiotherapy with concomitant and adjuvant temozolomide or temozolomide alone. As DNA-methyltransferases (DNMTs) are the enzymes responsible for setting up and maintaining DNA methylation patterns in eukaryotic cells, we analyzed further, whether MGMT promoter methylatio...

  16. 1-MCP处理结合冷藏对西兰花品质的影响%Effect of 1-MCP treatment on the quality of broccolis in cold storage

    Institute of Scientific and Technical Information of China (English)

    林本芳; 鲁晓翔; 李江阔; 陈绍慧

    2012-01-01

    This paper was conducted to investigated the effect of different concentrations (1.5, 2.5 μL/L) of 1-MCP on the quality of broccolis during 0℃ storage conditions. The physical and chemical indexes of broccolis were detected every 10 days. The results showed that: after treatment used 1-MCP, broccoli's soluble solids, Vc and chlorophyll content decreased less than ordinary storage. 1-MCP inhibited the increase of respiratory intensity, increased peroxidase(POD), superoxide dismutase(SOD) activities and slowed down polyphenol oxidase(PPO) activity. Additionally, treatment of 1-MCP also decreased the MDA content and cell membrane permeability, so delayed ripening of broccoli. 1-MCP treatment could delay ripening of broccoli, but 2.5 μL/L 1-MCP treatment is better than 1.5 μL/L treatment. Therefore, the storage effect of broccolis with 1-MCP treatment is superior to that of with only ordinary storage. 1-MCP still played normal function at low temperature.%研究了1—MCP处理对西兰花冷藏保鲜品质的影响。1-MCP处理浓度为1.5、2.5μL/L,冷藏期间每隔10d取样测定西兰花理化指标及营养成分。试验结果表明:与对照相比,1-MCP处理对于延缓西兰花可溶性固形物含量(TSS)、Vc含量及叶绿素含量的下降均有明显效果,并可抑制西兰花贮藏期间的呼吸强度。1-MCP处理提高西兰花过氧化物酶(POD)和超氧化物歧化酶(SOD)的活性,降低多酚氧化酶(PPO)活性,延缓了丙二醛(MDA)含量和细胞膜相对透性的升高,延缓了西兰花的衰老。1.5、2.5μL/L的1-MCP处理对西兰花的成熟衰老均有抑制作用,其中经2.5μL/L处理的西兰花的品质表现更好。这说明了冷藏结合1—MCP处理不仅较之普通冷藏更好地保持了西兰花的营养品质,同时也证明1—MCP在低温条件下仍可发挥正常作用。

  17. 30S Subunit-Dependent Activation of the Sorangium cellulosum So ce56 Aminoglycoside Resistance-Conferring 16S rRNA Methyltransferase Kmr

    Science.gov (United States)

    Savic, Miloje; Sunita, S.; Zelinskaya, Natalia; Desai, Pooja M.; Macmaster, Rachel; Vinal, Kellie

    2015-01-01

    Methylation of bacterial 16S rRNA within the ribosomal decoding center confers exceptionally high resistance to aminoglycoside antibiotics. This resistance mechanism is exploited by aminoglycoside producers for self-protection while functionally equivalent methyltransferases have been acquired by human and animal pathogenic bacteria. Here, we report structural and functional analyses of the Sorangium cellulosum So ce56 aminoglycoside resistance-conferring methyltransferase Kmr. Our results demonstrate that Kmr is a 16S rRNA methyltransferase acting at residue A1408 to confer a canonical aminoglycoside resistance spectrum in Escherichia coli. Kmr possesses a class I methyltransferase core fold but with dramatic differences in the regions which augment this structure to confer substrate specificity in functionally related enzymes. Most strikingly, the region linking core β-strands 6 and 7, which forms part of the S-adenosyl-l-methionine (SAM) binding pocket and contributes to base flipping by the m1A1408 methyltransferase NpmA, is disordered in Kmr, correlating with an exceptionally weak affinity for SAM. Kmr is unexpectedly insensitive to substitutions of residues critical for activity of other 16S rRNA (A1408) methyltransferases and also to the effects of by-product inhibition by S-adenosylhomocysteine (SAH). Collectively, our results indicate that adoption of a catalytically competent Kmr conformation and binding of the obligatory cosubstrate SAM must be induced by interaction with the 30S subunit substrate. PMID:25733511

  18. A SAM-dependent methyltransferase cotranscribed with arsenate reductase alters resistance to peptidyl transferase center-binding antibiotics in Azospirillum brasilense Sp7.

    Science.gov (United States)

    Singh, Sudhir; Singh, Chhaya; Tripathi, Anil Kumar

    2014-05-01

    The genome of Azospirillum brasilense harbors a gene encoding S-adenosylmethionine-dependent methyltransferase, which is located downstream of an arsenate reductase gene. Both genes are cotranscribed and translationally coupled. When they were cloned and expressed individually in an arsenate-sensitive strain of Escherichia coli, arsenate reductase conferred tolerance to arsenate; however, methyltransferase failed to do so. Sequence analysis revealed that methyltransferase was more closely related to a PrmB-type N5-glutamine methyltransferase than to the arsenate detoxifying methyltransferase ArsM. Insertional inactivation of prmB gene in A. brasilense resulted in an increased sensitivity to chloramphenicol and resistance to tiamulin and clindamycin, which are known to bind at the peptidyl transferase center (PTC) in the ribosome. These observations suggested that the inability of prmB:km mutant to methylate L3 protein might alter hydrophobicity in the antibiotic-binding pocket of the PTC, which might affect the binding of chloramphenicol, clindamycin, and tiamulin differentially. This is the first report showing the role of PrmB-type N5-glutamine methyltransferases in conferring resistance to tiamulin and clindamycin in any bacterium. PMID:24573606

  19. 不同质量浓度臭氧化水对鲜切西兰花贮藏品质的影响%Effect of Ozonated Water with Various Concentrations on Storage Quality of Fresh-cut Broccoli

    Institute of Scientific and Technical Information of China (English)

    王宏延; 曾凯芳; 贾凝; 陈存坤; 王文生

    2012-01-01

    采用不同质量浓度臭氧化水处理鲜切西兰花。结果表明:质量浓度2.0mg/L的臭氧化水处理组保鲜效果最优,在贮藏12d时与对照组相比,质量损失率降低24.37%,VC含量提高40.7%,多酚氧化酶酶活降低23.5%,过氧化物酶酶活降低25.6%;在4℃的条件下,质量浓度2.0mg/L的臭氧化水处理组鲜切西兰花贮藏期可达到15d。%Ozonated water can be used for disinfection,sterilization,deodorization and preservation with multiple advantages such as rapid sterilization rate,simple operation and without residues.It can be used directly for the sterilization of fresh-cut vegetables so that it is a potential technology of cold sterilization and green sterilization.The results showed that the best preservation effect on fresh-cut broccoli was observed at the condition of 2 mg/L ozonated water.Compared with the control,ozonated water-treated fresh-cut broccoli after storage for 12 days revealed a reduction by 24.37% for weight loss rate,an improvement by 40.7% for vitamin C,a decline by 23.5% for polyphenol oxidase activity and a decrease by 25.6% for peroxidase activity.The storage life of fresh-cut broccoli subjected to treatment with 2 mg/L ozonated water can be extended to 15 days at the storage temperature of 4 ℃.

  20. (Accumulation of methyl-deficient rat liver messenger ribonucleic acid on ethionine administration). Progress report. [Methyltransferase activity in Ehrlich ascites tumor cells and effects of phorbol ester on methyltransferase activity

    Energy Technology Data Exchange (ETDEWEB)

    Borek, E.

    1980-01-01

    Enzyme fractions were isolated from Ehrlich ascites cells which introduced methyl groups into methyl deficient rat liver mRNA and unmethylated vaccinia mRNA. The methyl groups were incorporated at the 5' end into cap 1 structures by the viral enzyme, whereas both cap 0 and cap 1 structures were formed by the Ehrlich ascites cell enzymes. Preliminary results indicate the presence of adenine N/sup 6/-methyltransferase activity in Ehrlich ascites cells. These results indicate that mRNA deficient in 5'-cap methylation and in internal methylation of adenine accumulated in rats on exposure to ethionine. The methyl-deficient mRNA isolated from the liver of ethionine-fed rats differed in its translational properties from mRNA isolated from control animals. Preliminary experiments indicate that single topical application of 17n moles of TPA to mouse skin altered tRNA methyltransferases. The extent of methylation was increased over 2-fold in mouse skin treated with TPA for 48 hours. These changes have been observed as early as 12 hours following TPA treatment. In contrast, the application of initiating dose of DMBA had no effect on these enzymes. It should be emphasized that the changes in tRNA methyltransferases produced by TPA are not merely an increase of the concentration of the enzyme, rather that they represent alterations of specificity of a battery of enzymes. In turn the change in enzyme specificity can produce alterations in the structure of tRNA. (ERB)

  1. The ankyrin repeats of G9a and GLP histone methyltransferases are mono- and dimethyllysine binding modules

    Energy Technology Data Exchange (ETDEWEB)

    Collins, Robert E.; Northrop, Jeffrey P.; Horton, John R.; Lee, David Y.; Zhang, Xing; Stallcup, Michael R.; Cheng, Xiaodong (USC); (Emory)

    2008-03-19

    Histone modifications have important roles in transcriptional control, mitosis and heterochromatin formation. G9a and G9a-like protein (GLP) are euchromatin-associated methyltransferases that repress transcription by mono- and dimethylating histone H3 at Lys9 (H3K9). Here we demonstrate that the ankyrin repeat domains of G9a and GLP bind with strong preference to N-terminal H3 peptides containing mono- or dimethyl K9. X-ray crystallography revealed the basis for recognition of the methylated lysine by a partial hydrophobic cage with three tryptophans and one acidic residue. Substitution of key residues in the cage eliminated the H3 tail interaction. Hence, G9a and GLP contain a new type of methyllysine binding module (the ankyrin repeat domains) and are the first examples of protein (histone) methyltransferases harboring in a single polypeptide the activities that generate and read the same epigenetic mark.

  2. A female adnexal tumor of probable Wolffian origin showing positive O-6-methylguanine-DNA methyltransferase methylation

    Science.gov (United States)

    Kwon, Min Jung; Yun, Min Jeong

    2016-01-01

    Female adnexal tumor of probable Wolffian origin (FATWO) is a rare disease entity that arises from the mesonephric duct system. FATWO is different than other gynecological cancers in terms of embryology. Here, we describe the case of a 52-year-old woman with malignant FATWO. The patient underwent explorative laparotomy and surgical staging after a frozen section revealed malignancy. Detailed examination of the pathologic findings were consistent with FATWO. Counseling and further testing were provided to the patient to assess the risk of germline mutation and epigenetic change. An O-6-methylguanine-DNA methyltransferase gene methylation test was positive, and all other tests were normal. This is the first study to report a case of O-6-methylguanine-DNA methyltransferase methylation with FATWO in Korea. PMID:27462603

  3. 西兰花多肽对大鼠C6胶质瘤细胞生长的影响%The effect of broccoli polypeptide on C6 glioma cells of rat

    Institute of Scientific and Technical Information of China (English)

    徐俊杰; 于洪泉; 国巍; 赵伟; 金宏; 温娜; 齐玲

    2012-01-01

    目的 研究西兰花多肽对大鼠C6胶质瘤细胞生长的影响.方法 培养C6胶质瘤细胞,用西兰花多肽(0、0.01、0.1、1、10μ/ml)作用细胞72h,MTT法检测胶质瘤细胞的生长情况.结果 西兰花多肽作用C6胶质瘤细胞72h时,各药物浓度组都表现为对细胞的抑制作用,1μl/ml组(0.58 ±0.08 vs 0.75 ±0.02)和10μl/ml组(0.07 ±0.01 vs 0.75 ±0.02)表现出显著的抑制生长作用(P<0.01).结论 西兰花多肽可抑制C6胶质瘤细胞的生长,并且随着药物浓度升高药物作用增强,说明西兰花多肽具有抑制肿瘤细胞生长的作用.%Objective To study the effects of broccoli polypeptide on the proliferation of C6 glioma cells. Method Cultured C6 glioma cells were treated with different dose of broccoli polypeptide(0,0. 01 ,0. 1 ,1 , 10μl/ml)for 72h. The level of cell growth was detected by MTT assay. Results After 72h,all dosed of polypeptide inhibited the growth of cells, especially in 1(0,1/μml group(0. 58 ±0.08 vs 0.75 ±0.02) and 10(1,1/μml group(0.07 ±0.01 vs 0.75 ±0.02) when compared with respective control group(P <0. 01). Conclusion Broccoli polypeptide may inhibit the proliferation of C6 glioma cells, and the effect is increased with dose, which suggests that broccoli polypeptide can inhibit the growth of tumor cells.

  4. 细胞分裂素类生长调节剂对青花菜采后衰老的影响%Effects of Cytokinin Growth Regulators on Senescence of Harvested Broccoli (Brassica oleracea L. var. italica) Florets

    Institute of Scientific and Technical Information of China (English)

    王云莉; 王成荣; 王然; 杨绍兰; 刘宪路

    2009-01-01

    Application of cytokinin growth regulators, KT (Kinetin), 6-BA (Benzyladenine), and CPPU [N-(2-chloro-4-pyridyl)-N'-pheuylurea] on delaying senescence of harvested broccoli florets was studied. The results showed that use of KT, 6-BA or CPPU on harvested broccoli delayed floret yellowing and chlorophyll degradation, decreased respiration and ethylene production rate, postponed climacteric time, and retarded membrane permeability loss. The content of endogenous ABA was lower and the levels of IAA and GA were higher in the broccoli treated with 6-BA and CPPU than those in the control during 12-84 h. The treated effects enhanced with increasing concentration in the study. The CPPU treatment (40 mg·L~(-1)) was most effective in delaying the broccoli yellowing, followed by 6-BA and KT.%以青花菜'绿皇后'品种为试材,研究了25℃条件下细胞分裂素类生长调节剂KT、6-BA和CPPU对延缓青花菜采后衰老的效果.结果表明:3种细胞分裂素类生长调节剂处理均在不同程度上延缓了青花菜花蕾黄化和叶绿素分解,推迟了呼吸和乙烯跃变峰出现的时间,降低了峰值,并维持了细胞膜稳定性.内源激素测定结果表明,6-BA和CPPU处理使青花菜花蕾在试验的前期阶段(12~84 h)一直保持相对较高的内源IAA和GA水平,相对较低的内源ABA水平.3种细胞分裂素类生长调节剂以CPPU延缓青花菜衰老效果最为明显,其次是6-BA,KT效果最弱.延缓青花菜采后衰老效果随着处理的浓度增加而增加,以40 mg·L~(-1)CPPU延缓青花菜衰老效果最佳.

  5. 不同西兰花品种中硫代葡萄糖苷的组分与含量分析%Glucosinolate Component and Content Analysis of Different Broccoli Varieties

    Institute of Scientific and Technical Information of China (English)

    丁云花; 何洪巨; 宋曙辉; 简元才; 赵学志; 王文琪

    2015-01-01

    Glucosinolates component and content of 29 broccoli varieties were analyzed by HPLC method. As a result, a total of eight kinds of glucosinolates in broccoli bud-head were detected. Four kinds of them were aliphatic glucosino-lates, such as glucoiberin(IBE), progoitrin(PRO), glucoraphanin(RAA) and gluconapin(NAP), and the others were indolic glucosinolates, such as 4-Hydroxyglucobrassicin(4OH), glucobrassicin(GBC), 4-Methoxyglucobrassicin(4ME) and Neoglu-cobrassicin (NEO). The content of total glucosinolates was 11.27μmol·g-1 DW. RAA, GBC and NEO were the predomi-nant component of glucosinolates in broccoli bud-head, with a proportion of 34.78%, 25.02% and 23.25% respectively. The content of total glucosinolates in different variety of broccoli varied from 5.60 μmol·g-1 DW to 17.04 μmol·g-1 DW.%采用 HPLC 法测定分析了29个西兰花品种的硫代葡萄糖苷组分与含量,结果检测到8种硫代葡萄糖苷,包括4种脂肪族硫苷:3-甲基硫氧烯丙基硫苷、2-羟基-3-丁烯基硫苷、4-甲基硫氧丁基硫苷和3-丁烯基硫苷;4种吲哚族硫苷:4-羟基吲哚基-3-甲基硫苷、3-甲基吲哚基硫苷、4-甲氧基吲哚基-3-甲基硫苷和1-甲氧基吲哚基-3-甲基硫苷。总硫苷含量平均为11.27μmol/g,RAA、GBC 和 NEO 是西兰花三种主要的硫苷成分,分别占总硫苷含量的34.78%、25.02%和23.25%。不同西兰花品种的总硫苷含量变异范围5.60~17.04μmol/g,存在较大差异。

  6. Control Efficacy of Insecticides Mixture on Broccolis%几种药剂混配防治西兰花根部病害综合效果试验

    Institute of Scientific and Technical Information of China (English)

    方辉; 王会福; 余山红; 毛肇蓬

    2013-01-01

      为了有效防治西兰花根肿病、根腐病、根结线虫病等根部病害,提升西兰花的产量和品质,采用灌根法进行几种药剂混配防治西兰花根部病害田间药效试验,结果表明:68%精甲霜•锰锌WG 500倍+3.2%阿维菌素EC 1000倍+微补精力600倍+微补根力钙600倍对西兰花根部病害具有较好的综合防治效果,其中连续2次灌根后50 d的校正防效达82.1%,分别比68%精甲霜•锰锌WG 500倍+3.2%阿维菌素EC 1000倍、5%石灰氮GR 1200 kg/hm2高出5.6%、21.3%,持效期较长,安全性好,因此可以在生产上加以推广应用。%  The purpose of the trial was to control clubroot disease, ceitocybe bescens, root knot nematode disease of broccoli, in order to promote the broccoli production and quality. Mixture of several insecticides was used on broccoli by root­irrigation method in this trail. The result showed that the mixture of 68%Mefenoxam­Mancozeb WG 1:500 plus 3.2%Avermectins EC 1:1 000 plus 425 g/L MNPK­38 g/L Calcium 1:600 and 160 g/L Calcium 1:600 had better control effect on Broccoli root diseases, and the 50 days’ correction control effect after two consecutive root­pouring reached 82.1%, which was higher than that of the mixture of 68%Mefenoxam­Mancozeb WG 1:500 plus 3.2%Avermectins EC 1:1 000 and that of 5%Calcium Cyanamide GR 1 200kg/hm2 by 5.6% and 21.3%, respectively. With longer effective lasting period and better security, this mixture can be demonstrated to apply to the practical production.

  7. Structure of the Q237W mutant of HhaI DNA methyltransferase: an insight into protein-protein interactions

    OpenAIRE

    Dong, Aiping; Zhou, Lan; Zhang, Xing; Stickel, Shawn; Roberts, Richard J.; Cheng, Xiaodong

    2004-01-01

    We have determined the structure of a mutant (Q237W) of HhaI DNA methyltransferase, complexed with the methyl-donor product AdoHcy. The Q237W mutant proteins were crystallized in the monoclinic space group C2 with two molecules in the crystallographic asymmetric unit. Protein-protein interface calculations in the crystal lattices suggest that the dimer interface has the specific characteristics for homodimer protein-protein interactions, while the two active sites are spatially...

  8. O6-methylguanine DNA methyltransferase as a promising target for the treatment of temozolomide-resistant gliomas

    OpenAIRE

    Fan, C-H; Liu, W-L; Cao, H.; Wen, C; Chen, L.; G. Jiang

    2013-01-01

    Temozolomide (TMZ) is an alkylating agent currently used as first-line therapy for gliomas treatment due to its DNA-damaging effect. However, drug resistance occurs, preventing multi-cycle use of this chemotherapeutic agent. One of the major mechanisms of cancer drug resistance is enhanced activity of a DNA repair enzyme, O6-methylguanine-DNA-methyltransferase (MGMT), which counteracts chemotherapy-induced DNA alkylation and is a key component of chemoresistance. MGMT repairs TMZ-induced DNA ...

  9. Cytoplasmic sequestration of an O6-methylguanine-DNA methyltransferase enhancer binding protein in DNA repair-deficient human cells

    OpenAIRE

    Frank Y. Chen; Harris, Linda C.; Joanna S Remack; Brent, Thomas P.

    1997-01-01

    O6-Methylguanine-DNA methyltransferase (MGMT), an enzyme that repairs adducts at O6 of guanine in DNA, is a major determinant of susceptibility to simple methylating carcinogens or of tumor response to anticancer chloroethylating drugs. To investigate the mechanisms underlying cellular expression of this DNA repair enzyme, we focused on the role of a 59-bp enhancer of the human MGMT gene in the regulation of its expression. By using chloramphenicol acetyltransferase reporter assays, we found ...

  10. A novel route to product specificity in the Suv4-20 family of histone H4K20 methyltransferases

    OpenAIRE

    Southall, Stacey M.; Cronin, Nora B.; Wilson, Jon R.

    2013-01-01

    The delivery of site-specific post-translational modifications to histones generates an epigenetic regulatory network that directs fundamental DNA-mediated processes and governs key stages in development. Methylation of histone H4 lysine-20 has been implicated in DNA repair, transcriptional silencing, genomic stability and regulation of replication. We present the structure of the histone H4K20 methyltransferase Suv4-20h2 in complex with its histone H4 peptide substrate and S-adenosyl methion...

  11. Effects of Active‐Site Modification and Quaternary Structure on the Regioselectivity of Catechol‐O‐Methyltransferase

    OpenAIRE

    Law, Brian J. C.; Bennett, Matthew R.; Thompson, Mark L; Levy, Colin; Shepherd, Sarah A; Leys, David; Micklefield, Jason

    2016-01-01

    Abstract Catechol‐O‐methyltransferase (COMT), an important therapeutic target in the treatment of Parkinson's disease, is also being developed for biocatalytic processes, including vanillin production, although lack of regioselectivity has precluded its more widespread application. By using structural and mechanistic information, regiocomplementary COMT variants were engineered that deliver either meta‐ or para‐methylated catechols. X‐ray crystallography further revealed how the active‐site r...

  12. Noncompetitive Inhibition of Indolethylamine-N-methyltransferase by N,N-Dimethyltryptamine and N,N-Dimethylaminopropyltryptamine

    OpenAIRE

    Chu, Uyen B.; Vorperian, Sevahn K.; Satyshur, Kenneth; Eickstaedt, Kelsey; Cozzi, Nicholas V; Mavlyutov, Timur; Hajipour, Abdol R.; Ruoho, Arnold E.

    2014-01-01

    Indolethylamine-N-methyltransferase (INMT) is a Class 1 transmethylation enzyme known for its production of N,N-dimethyltryptamine (DMT), a hallucinogen with affinity for various serotonergic, adrenergic, histaminergic, dopaminergic, and sigma-1 receptors. DMT is produced via the action of INMT on the endogenous substrates tryptamine and S-adenosyl-l-methionine (SAM). The biological, biochemical, and selective small molecule regulation of INMT enzyme activity remain largely unknown. Kinetic m...

  13. Protein Isoaspartate Methyltransferase Prevents Apoptosis Induced by Oxidative Stress in Endothelial Cells: Role of Bcl-Xl Deamidation and Methylation

    OpenAIRE

    Amelia Cimmino; Rosanna Capasso; Fabbri Muller; Irene Sambri; Lucia Masella; Marianna Raimo; Maria Luigia De Bonis; Stefania D'Angelo; Vincenzo Zappia; Patrizia Galletti; Diego Ingrosso

    2008-01-01

    BACKGROUND: Natural proteins undergo in vivo spontaneous post-biosynthetic deamidation of specific asparagine residues with isoaspartyl formation. Deamidated-isomerized molecules are both structurally and functionally altered. The enzyme isoaspartyl protein carboxyl-O-methyltransferase (PCMT; EC 2.1.1.77) has peculiar substrate specificity towards these deamidated proteins. It catalyzes methyl esterification of the free alpha-carboxyl group at the isoaspartyl site, thus initiating the repair ...

  14. Genome-wide identification and comparative analysis of cytosine-5 DNA methyltransferases and demethylase families in wild and cultivated peanut

    Directory of Open Access Journals (Sweden)

    Pengfei eWang

    2016-02-01

    Full Text Available AbstractDNA methylation plays important roles in genome protection, regulation of gene expression and was associated with plants development. Plant DNA methylation pattern was mediated by cytosine-5 DNA methyltransferases and demethylase. Although the genomes of AA and BB wild peanuts have been fully sequence, these two gene families have not been studied. In this study we report the identification and analysis of putative cytosine-5 DNA methyltransferases (C5-MTases and demethylase in AA and BB wild peanuts. Cytosine-5 DNA methyltransferases in AA and BB wild peanuts could be classified in known MET, CMT and DRM2 groups based on their domain organization. This result was supported by the gene and protein structural characteristics and phylogenetic analysis. We found that some wild peanut DRM2 numbers didn’t contain UBA domain which was different from other plants such as Arabidopsis, maize, soybean. Five DNA demethylase were found in AA genome and five in BB genome. The selective pressure analysis showed that wild peanut C5-MTases gene mainly underwent purifying selection but many positive selection sites can be detected. Conversely, DNA demethylase genes mainly underwent positive selection during evolution. Additionally, the expression dynamic of cytosine-5 DNA methyltransferases and demethylase genes in different cultivated peanut tissues were analyzed. Expression result showed that cold, heat or drought stress could influence the expression level of C5-MTases and DNA demethylase genes in cultivated peanut. These results are useful for better understanding the complexity of these two gene families, and will facilitate epigenetic studies in peanut.

  15. O6-Methylguanine-DNA methyltransferase status in neuroendocrine tumours: prognostic relevance and association with response to alkylating agents

    OpenAIRE

    Walter, T.; van Brakel, B; Vercherat, C; Hervieu, V; Forestier, J.; Chayvialle, J-A; Molin, Y.; Lombard-Bohas, C; Joly, M-O; Scoazec, J-Y

    2015-01-01

    Background: O6-Methylguanine-DNA methyltransferase (MGMT) loss of expression has been suggested to be predictive of response to temozolomide in neuroendocrine tumours (NETs), but so far, only limited data are available. We evaluated the prognostic and predictive value of MGMT status, assessed by two molecular methods and immunohistochemistry, in a large series of NETs of different origins. Methods: A total of 107 patients, including 53 treated by alkylants (temozolomide, dacarbazine or strept...

  16. Methylation pattern of the O6-methylguanine-DNA methyltransferase gene in colon during progressive colorectal tumorigenesis

    OpenAIRE

    NAGASAKA, Takeshi; Goel, Ajay; Notohara, Kenji; Takahata, Takaomi; Sasamoto, Hiromi; Uchida, Takuyuki; Nishida, Naoshi; Tanaka, Noriaki; Boland, Clement Richard; Matsubara, Nagahide

    2008-01-01

    O6-methylguanine-DNA methyltransferase (MGMT) is a DNA repair gene which is frequently methylated in colorectal cancer (CRC). However, it remains controversial whether methylation of specific CpG sequences within MGMT promoter leads to loss of its protein expression, and if MGMT methylation correlates with G to A transition mutations in KRAS. Two methylation sensitive regions (Mp and Eh region) of MGMT promoter were investigated in 593 specimens of colorectal tissue: 233 CRCs, 104 adenomatous...

  17. O6-Methylguanine DNA methyltransferase protein expression in tumor cells predicts outcome of temozolomide therapy in glioblastoma patients

    OpenAIRE

    Spiegl-Kreinecker, Sabine; Pirker, Christine; Filipits, Martin; Lötsch, Daniela; Buchroithner, Johanna; Pichler, Josef; Silye, Rene; Weis, Serge; Micksche, Michael; Fischer, Johannes; Berger, Walter

    2009-01-01

    O6-Methylguanine DNA methyltransferase (MGMT) is implicated as a major predictive factor for treatment response to alkylating agents including temozolomide (TMZ) of glioblastoma multiforme (GBM) patients. However, whether the MGMT status in GBM patients should be detected at the level of promoter methylation or protein expression is still a matter of debate. Here, we compared promoter methylation (by methylation-specific polymerase chain reaction) and protein expression (by Western blot) in t...

  18. A Small-Molecule Probe of the Histone Methyltransferase G9a Induces Cellular Senescence in Pancreatic Adenocarcinoma

    OpenAIRE

    Wang, Qiu; Paulk, Joshiawa Lanair James; Kubicek, Stefan; Kemp, Melissa M.; Adams, Drew J.; Shamji, Alykhan Farid; Wagner, Bridget K.; Schreiber, Stuart L.; Yuan, Yuan

    2012-01-01

    Post-translational modifications of histones alter chromatin structure and play key roles in gene expression and specification of cell states. Small molecules that target chromatin-modifying enzymes selectively are useful as probes and have promise as therapeutics, although very few are currently available. G9a (also named euchromatin histone methyltransferase 2 (EHMT2)) catalyzes methylation of lysine 9 on histone H3 (H3K9), a modification linked to aberrant silencing of tumor-suppressor gen...

  19. Downregulation of Histone H3 Lysine 9 Methyltransferase G9a Induces Centrosome Disruption and Chromosome Instability in Cancer Cells

    OpenAIRE

    Kondo, Yutaka; Shen, Lanlan; Ahmed, Saira; Boumber, Yanis; Sekido, Yoshitaka; Haddad, Bassem R; Issa, Jean-Pierre J.

    2008-01-01

    Background Modifications of the histone amino-terminal tails affect access of regulatory factors and complexes to chromatin and thereby influence biological processes. Cancer cells are characterized by prominent epigenetic dysregulation, including histone modifications. However, the functional roles of the histone methyltransferases (HMT) in cancer remain unclear. Methodology/Principal Findings We studied RNAi-based inhibition (knockdown, KD) of 2 different H3K9 HMTs, SUV39H1 and G9a. Knockdo...

  20. Activating and inhibitory functions for the histone lysine methyltransferase G9a in T helper cell differentiation and function

    OpenAIRE

    Lehnertz, Bernhard; Northrop, Jeffrey P.; Antignano, Frann; Burrows, Kyle; Hadidi, Sima; Mullaly, Sarah C.; Rossi, Fabio M.V.; Zaph, Colby

    2010-01-01

    Accumulating evidence suggests that the regulation of gene expression by histone lysine methylation is crucial for several biological processes. The histone lysine methyltransferase G9a is responsible for the majority of dimethylation of histone H3 at lysine 9 (H3K9me2) and is required for the efficient repression of developmentally regulated genes during embryonic stem cell differentiation. However, whether G9a plays a similar role in adult cells is still unclear. We identify a critical role...

  1. Epistasis between catechol-O-methyltransferase and type II metabotropic glutamate receptor 3 genes on working memory brain function

    OpenAIRE

    Tan, Hao-Yang; Chen, Qiang; Sust, Steven; Joshua W Buckholtz; Meyers, John D.; Egan, Michael F.; Mattay, Venkata S.; Meyer-Lindenberg, Andreas; Weinberger, Daniel R.; Callicott, Joseph H.

    2007-01-01

    Dopaminergic and glutamatergic systems are critical components responsible for prefrontal signal-to-noise tuning in working memory. Recent functional MRI (fMRI) studies of genetic variation in these systems in catechol-O-methyltransferase (COMT) and in metabotropic glutamate receptor mgluR3 (GRM3), respectively, suggest that these genes influence prefrontal physiological signal-to-noise in humans. Here, using fMRI, we extend these individual gene findings to examine the combined effects of CO...

  2. Therapy and progression – induced O6-methylguanine-DNA methyltransferase and mismatch repair alterations in recurrent glioblastoma multiforme

    OpenAIRE

    Agarwal, S.; Suri, V.; M C Sharma; C. Sarkar

    2015-01-01

    Despite multimodality treatment protocol including surgical resection, radiotherapy, and chemotherapy in patients with glioblastoma multiforme (GBM), most suffer from treatment failure and tumor recurrence within a few months of initial surgery. The effectiveness of temozolomide (TMZ), the most commonly used chemotherapeutic agent, is largely dependent on the methylation status of the promoter of the gene O6-methylguanine-DNA methyltransferase (MGMT) and the integrity of the mismatch repair (...

  3. Affect-Modulated Startle: Interactive Influence of Catechol-O-Methyltransferase Val158Met Genotype and Childhood Trauma

    OpenAIRE

    Klauke, Benedikt; Winter, Bernward; Gajewska, Agnes; Zwanzger, Peter; Reif, Andreas; Herrmann, Martin J.; Dlugos, Andrea; Warrings, Bodo; Jacob, Christian; Mühlberger, Andreas; Arolt, Volker; Pauli, Paul; Deckert, Jürgen; Domschke, Katharina

    2012-01-01

    The etiology of emotion-related disorders such as anxiety or affective disorders is considered to be complex with an interaction of biological and environmental factors. Particular evidence has accumulated for alterations in the dopaminergic and noradrenergic system – partly conferred by catechol-O-methyltransferase (COMT) gene variation – for the adenosinergic system as well as for early life trauma to constitute risk factors for those conditions. Applying a multi-level approach, in a sample...

  4. Catechol O-Methyltransferase Haplotype Predicts Immediate Musculoskeletal Neck Pain and Psychological Symptoms after Motor Vehicle Collision

    OpenAIRE

    McLean, Samuel A.; Diatchenko, Luda; Lee, Young M.; Swor, Robert A.; Domeier, Robert M; Jones, Jeffrey S.; Jones, Christopher W.; Reed, Caroline; Harris, Richard E; Maixner, William; Clauw, Daniel J.; Liberzon, Israel

    2010-01-01

    Genetic variations in the catechol-o-methyltransferase (COMT) gene have been associated with experimental pain and risk of chronic pain development, but no studies have examined genetic predictors of neck pain intensity and other patient characteristics after motor vehicle collision (MVC). We evaluated the association between COMT genotype and acute neck pain intensity and other patient characteristics in 89 Caucasian individuals presenting to the emergency department (ED) after MVC. In the E...

  5. Investigating the Potential Role of Genetic and Epigenetic Variation of DNA Methyltransferase Genes in Hyperplastic Polyposis Syndrome

    OpenAIRE

    Drini, Musa; Nicholas C. Wong; Hamish S Scott; Craig, Jeffrey M; Dobrovic, Alexander; Chelsee A Hewitt; Dow, Christofer; Young, Joanne P; Jenkins, Mark A; Saffery, Richard; Macrae, Finlay A.

    2011-01-01

    Background Hyperplastic Polyposis Syndrome (HPS) is a condition associated with multiple serrated polyps, and an increased risk of colorectal cancer (CRC). At least half of CRCs arising in HPS show a CpG island methylator phenotype (CIMP), potentially linked to aberrant DNA methyltransferase (DNMT) activity. CIMP is associated with methylation of tumor suppressor genes including regulators of DNA mismatch repair (such as MLH1, MGMT), and negative regulators of Wnt signaling (such as WIF1). In...

  6. Methyltransferase PRMT1 Is a Binding Partner of HBx and a Negative Regulator of Hepatitis B Virus Transcription

    OpenAIRE

    Benhenda, Shirine; Ducroux, Aurélie; Rivière, Lise; Sobhian, Bijan; Ward, Michael D.; Dion, Sarah; Hantz, Olivier; Protzer, Ulrike; Michel, Marie-Louise; Benkirane, Monsef; Semmes, Oliver J.; Buendia, Marie-Annick; Neuveut, Christine

    2013-01-01

    The hepatitis B virus X protein (HBx) is essential for virus replication and has been implicated in the development of liver cancer. HBx is recruited to viral and cellular promoters and activates transcription by interacting with transcription factors and coactivators. Here, we purified HBx-associated factors in nuclear extracts from HepG2 hepatoma cells and identified protein arginine methyltransferase 1 (PRMT1) as a novel HBx-interacting protein. We showed that PRMT1 overexpression reduced ...

  7. Direct methylation of FXR by Set7/9, a lysine methyltransferase, regulates the expression of FXR target genes

    OpenAIRE

    Balasubramaniyan, Natarajan; Ananthanarayanan, Meena; Suchy, Frederick J.

    2012-01-01

    The farnesoid X receptor (FXR) is a ligand (bile acid)-dependent nuclear receptor that regulates target genes involved in every aspect of bile acid homeostasis. Upon binding of ligand, FXR recruits an array of coactivators and associated proteins, some of which have intrinsic enzymatic activity that modify histones or even components of the transcriptional complex. In this study, we show chromatin occupancy by the Set7/9 methyltransferase at the FXR response element (FXRE) and direct methylat...

  8. The Role of the Methyltransferase Domain of Bifunctional Restriction Enzyme RM.BpuSI in Cleavage Activity

    OpenAIRE

    Arthur Sarrade-Loucheur; Shuang-yong Xu; Siu-Hong Chan

    2013-01-01

    Restriction enzyme (REase) RM.BpuSI can be described as a Type IIS/C/G REase for its cleavage site outside of the recognition sequence (Type IIS), bifunctional polypeptide possessing both methyltransferase (MTase) and endonuclease activities (Type IIC) and endonuclease activity stimulated by S-adenosyl-L-methionine (SAM) (Type IIG). The stimulatory effect of SAM on cleavage activity presents a major paradox: a co-factor of the MTase activity that renders the substrate unsusceptible to cleavag...

  9. The Putative Protein Methyltransferase LAE1 of Trichoderma atroviride Is a Key Regulator of Asexual Development and Mycoparasitism

    OpenAIRE

    Aghcheh, Razieh Karimi; Irina S. Druzhinina; Kubicek, Christian P

    2013-01-01

    In Ascomycota the protein methyltransferase LaeA is a global regulator that affects the expression of secondary metabolite gene clusters, and controls sexual and asexual development. The common mycoparasitic fungus Trichoderma atroviride is one of the most widely studied agents of biological control of plant-pathogenic fungi that also serves as a model for the research on regulation of asexual sporulation (conidiation) by environmental stimuli such as light and/or mechanical injury. In order ...

  10. Genetic contribution of catechol-O-methyltransferase variants in treatment outcome of low back pain: a prospective genetic association study

    OpenAIRE

    Omair, Ahmad; Benedicte A Lie; Reikeras, Olav; Holden, Marit; Brox, Jens I

    2012-01-01

    Background Treatment outcome of low back pain (LBP) is associated with inter-individual variations in pain relief and functional disability. Genetic variants of catechol-O-methyltransferase (COMT) gene have previously been shown to be associated with pain sensitivity and pain medication. This study examines the association between COMT polymorphisms and 7–11 year change in Oswestry Disability Index (ODI) and Visual Analog Score (VAS) for LBP as clinical outcome variables in...

  11. Genetic contribution of catechol-O-methyltransferase variants in treatment outcome of low back pain: a prospective genetic association study

    OpenAIRE

    Omair Ahmad; Lie Benedicte; Reikeras Olav; Holden Marit; Brox Jens

    2012-01-01

    Abstract Background Treatment outcome of low back pain (LBP) is associated with inter-individual variations in pain relief and functional disability. Genetic variants of catechol-O-methyltransferase (COMT) gene have previously been shown to be associated with pain sensitivity and pain medication. This study examines the association between COMT polymorphisms and 7–11 year change in Oswestry Disability Index (ODI) and Visual Analog Score (VAS) for LBP as clinical outcome variables in patients ...

  12. Genetic contribution of catechol-O-methyltransferase variants in treatment outcome of low back pain: a prospective genetic association study

    OpenAIRE

    Omair, Ahmad; Lie, Benedicte Alexandra; Reikeras, Olav; Holden, Marit; Brox, Jens Ivar

    2012-01-01

    Background Treatment outcome of low back pain (LBP) is associated with inter-individual variations in pain relief and functional disability. Genetic variants of catechol-O-methyltransferase (COMT) gene have previously been shown to be associated with pain sensitivity and pain medication. This study examines the association between COMT polymorphisms and 7–11 year change in Oswestry Disability Index (ODI) and Visual Analog Score (VAS) for LBP as clinical outcome variables in patients treated w...

  13. The histone methyltransferase SDG8 mediates the epigenetic modification of light and carbon responsive genes in plants

    OpenAIRE

    Ying LI; Mukherjee, Indrani; Thum, Karen E; Tanurdzic, Milos; Katari, Manpreet S.; Obertello, Mariana; Edwards, Molly B; McCombie, W Richard; Martienssen, Robert A.; Coruzzi, Gloria M.

    2015-01-01

    Background Histone methylation modifies the epigenetic state of target genes to regulate gene expression in the context of developmental and environmental changes. Previously, we used a positive genetic screen to identify an Arabidopsis mutant, cli186, which was impaired in carbon and light signaling. Here, we report a deletion of the Arabidopsis histone methyltransferase SDG8 in this mutant (renamed sdg8-5), which provides a unique opportunity to study the global function of a specific histo...

  14. MTAP deletion confers enhanced dependency on the PRMT5 arginine methyltransferase in cancer cells | Office of Cancer Genomics

    Science.gov (United States)

    The discovery of cancer dependencies has the potential to inform therapeutic strategies and to identify putative drug targets. Integrating data from comprehensive genomic profiling of cancer cell lines and from functional characterization of cancer cell dependencies, we discovered that loss of the enzyme methylthioadenosine phosphorylase (MTAP) confers a selective dependence on protein arginine methyltransferase 5 (PRMT5) and its binding partner WDR77. MTAP is frequently lost due to its proximity to the commonly deleted tumor suppressor gene, CDKN2A.

  15. The Histone Methyltransferase SMYD2 Methylates PARP1 and Promotes Poly(ADP-ribosyl)ation Activity in Cancer Cells

    OpenAIRE

    Lianhua Piao; Daechun Kang; Takehiro Suzuki; Akiko Masuda; Naoshi Dohmae; Yusuke Nakamura; Ryuji Hamamoto

    2014-01-01

    Poly(ADP-ribose) polymerase-1 (PARP1) catalyzes the poly(ADP-ribosyl)ation of protein acceptors using NAD+ as the substrate is now considered as an important target for development of anticancer therapy. PARP1 is known to be post-translationally modified in various ways including phosphorylation and ubiquitination, but the physiological role of PARP1 methylation is not well understood. Herein we demonstrated that the histone methyltransferase SMYD2, which plays critical roles in human carcino...

  16. The Histone Methyltransferase SMYD2 Methylates PARP1 and Promotes Poly(ADP-ribosyl)ation Activity in Cancer Cells12

    OpenAIRE

    Piao, Lianhua; Kang, Daechun; Suzuki, Takehiro; Masuda, Akiko; Dohmae, Naoshi; Nakamura, Yusuke; Hamamoto, Ryuji

    2014-01-01

    Poly(ADP-ribose) polymerase-1 (PARP1) catalyzes the poly(ADP-ribosyl)ation of protein acceptors using NAD+ as the substrate is now considered as an important target for development of anticancer therapy. PARP1 is known to be post-translationally modified in various ways including phosphorylation and ubiquitination, but the physiological role of PARP1 methylation is not well understood. Herein we demonstrated that the histone methyltransferase SMYD2, which plays critical roles in human carcino...

  17. DNA methyltransferase 1 functions through C/ebpa to maintain hematopoietic stem and progenitor cells in zebrafish

    OpenAIRE

    Liu, Xiaohui; Jia, Xiaoe; Yuan, Hao; Ma, Ke; Chen, Yi; Jin, Yi; Deng, Min; Pan, Weijun; Chen, Saijuan; Chen, Zhu; de The, Hugues; Zon, Leonard; Zhou, Yi; Zhou, Jun; Zhu, Jun

    2015-01-01

    Background: DNA methyltransferase 1 (Dnmt1) regulates expression of many critical genes through maintaining parental DNA methylation patterns on daughter DNA strands during mitosis. It is essential for embryonic development and diverse biological processes, including maintenance of hematopoietic stem and progenitor cells (HSPCs). However, the precise molecular mechanism of how Dnmt1 is involved in HSPC maintenance remains unexplored. Methods: An N-ethyl-N-nitrosourea (ENU)-based genetic scree...

  18. The Histone Methyltransferase Inhibitor A-366 Uncovers a Role for G9a/GLP in the Epigenetics of Leukemia.

    Directory of Open Access Journals (Sweden)

    William N Pappano

    Full Text Available Histone methyltransferases are epigenetic regulators that modify key lysine and arginine residues on histones and are believed to play an important role in cancer development and maintenance. These epigenetic modifications are potentially reversible and as a result this class of enzymes has drawn great interest as potential therapeutic targets of small molecule inhibitors. Previous studies have suggested that the histone lysine methyltransferase G9a (EHMT2 is required to perpetuate malignant phenotypes through multiple mechanisms in a variety of cancer types. To further elucidate the enzymatic role of G9a in cancer, we describe herein the biological activities of a novel peptide-competitive histone methyltransferase inhibitor, A-366, that selectively inhibits G9a and the closely related GLP (EHMT1, but not other histone methyltransferases. A-366 has significantly less cytotoxic effects on the growth of tumor cell lines compared to other known G9a/GLP small molecule inhibitors despite equivalent cellular activity on methylation of H3K9me2. Additionally, the selectivity profile of A-366 has aided in the discovery of a potentially important role for G9a/GLP in maintenance of leukemia. Treatment of various leukemia cell lines in vitro resulted in marked differentiation and morphological changes of these tumor cell lines. Furthermore, treatment of a flank xenograft leukemia model with A-366 resulted in growth inhibition in vivo consistent with the profile of H3K9me2 reduction observed. In summary, A-366 is a novel and highly selective inhibitor of G9a/GLP that has enabled the discovery of a role for G9a/GLP enzymatic activity in the growth and differentiation status of leukemia cells.

  19. Association Between the Catechol-O-Methyltransferase Val158Met Polymorphism and Self-Perceived Social Acceptance in Adolescent Girls

    OpenAIRE

    Waugh, Christian E.; Dearing, Karen F.; Joormann, Jutta; GOTLIB, IAN H.

    2009-01-01

    Low perceived social acceptance is a significant risk factor for emotional difficulties in children. No studies, however, have examined genetic factors that may underlie individual differences in perceived social acceptance. In the present study we examined the relation between polymorphisms on the catechol-O-methyltransferase (COMT) Val158Met and serotonin transporter promoter (5-HTTLPR) genes and perceived social acceptance in 103 adolescent girls. Only the COMT polymorphism was related to ...

  20. Association between the Catechol-O-Methyltransferase (COMT) Val158Met Polymorphism and Manual Aiming Control in Healthy Subjects

    OpenAIRE

    Lage, Guilherme M.; Miranda, Débora M.; Romano-Silva, Marco A.; Simone B. Campos; Albuquerque, Maicon R.; Corrêa, Humberto; Leandro F. Malloy-Diniz

    2014-01-01

    Background Prefrontal dopamine is catabolized by the catechol-O-methyltransferase (COMT) enzyme. Current evidence suggests that the val/met single nucleotide polymorphism in the COMT gene can predict the efficiency of executive cognition in humans. Individuals carrying the val allele perform more poorly because less synaptic dopamine is available. Methodology/Principal Findings We investigated the influence of the COMT polymorphism on motor performance in a task that requires different execut...