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Academic literature on the topic 'HAT activity'

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Published: 10 December 2022
Last updated: 28 January 2023

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Journal articles on the topic "HAT activity"

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Yang, Hongmei, Wei Wei, Michael Menconi, and Per-Olof Hasselgren. "Dexamethasone-induced protein degradation in cultured myotubes is p300/HAT dependent." American Journal of Physiology-Regulatory, Integrative and Comparative Physiology 292, no. 1 (January 2007): R337—R334. http://dx.doi.org/10.1152/ajpregu.00230.2006.

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Muscle proteolysis during sepsis and other catabolic conditions is, at least in part, regulated by glucocorticoids. Dexamethasone-treated myotubes are a commonly used in vitro model of muscle wasting. We reported recently that treatment of cultured L6 myotubes with dexamethasone resulted in increased gene and protein expression of the nuclear cofactor p300 but it is not known whether glucocorticoids upregulate p300 histone acetyl transferase (HAT) activity in muscle and whether p300/HAT activity regulates glucocorticoid-induced muscle proteolysis. Here, we found that treatment of cultured L6 myotubes with dexamethasone resulted in increased nuclear p300/HAT activity. Treatment of myotubes with p300 siRNA or transfection of muscle cells with a plasmid expressing p300 that was mutated in its HAT activity domain blocked the dexamethasone-induced increase in protein degradation, supporting a role of p300/HAT in glucocortiocoid-induced muscle proteolysis. In addition to increased HAT activity, treatment of the myotubes with dexamethasone resulted in reduced nuclear expression and activity of histone deacetylases (HDACs) 3 and 6. When myotubes were treated with the HDAC inhibitor trichostatin A, protein degradation increased to the same degree as in dexamethasone-treated myotubes. The results suggest that glucocorticoids increase HAT and decrease HDAC activities in muscle, changes that both favor hyperacetylation. The results also provide evidence that dexamethasone-induced protein degradation in cultured myotubes is, at least in part, regulated by p300/HAT activity.
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Smith, Aaron T., Meredith R. Livingston, Antonello Mai, Patrizia Filetici, Sherry F. Queener, and William J. Sullivan. "Quinoline Derivative MC1626, a Putative GCN5 Histone Acetyltransferase (HAT) Inhibitor, Exhibits HAT-Independent Activity against Toxoplasma gondii." Antimicrobial Agents and Chemotherapy 51, no. 3 (December 18, 2006): 1109–11. http://dx.doi.org/10.1128/aac.01256-06.

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ABSTRACT We report that quinoline derivative MC1626, first described as an inhibitor of the histone acetyltransferase (HAT) GCN5, is active against the protozoan parasite Toxoplasma gondii in vitro. However, MC1626 does not inhibit Toxoplasma GCN5 HATs or reduce HAT-mediated activity; rather, this quinoline may target the plastid organelle called the apicoplast.
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Hansson, Magnus L., Anita E. Popko-Ścibor, Mariana Saint Just Ribeiro, Beverley M. Dancy, Mikael J. Lindberg, Philip A. Cole, and Annika E. Wallberg. "The transcriptional coactivator MAML1 regulates p300 autoacetylation and HAT activity." Nucleic Acids Research 37, no. 9 (March 20, 2009): 2996–3006. http://dx.doi.org/10.1093/nar/gkp163.

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Hsu, Chih-Hung, Margaret D. T. Chang, Kang-Yu Tai, Yu-Ting Yang, Pei-Shan Wang, Chi-Ju Chen, Yan-Hsiung Wang, Sheng-Chung Lee, Cheng-Wen Wu, and Li-Jung Juan. "HCMV IE2-mediated inhibition of HAT activity downregulates p53 function." EMBO Journal 23, no. 11 (May 13, 2004): 2269–80. http://dx.doi.org/10.1038/sj.emboj.7600239.

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Menzel, Gerhard, Tony Heitkam, Kathrin M. Seibt, Faisal Nouroz, Manuela Müller-Stoermer, John S. Heslop-Harrison, and Thomas Schmidt. "The diversification and activity of hAT transposons in Musa genomes." Chromosome Research 22, no. 4 (November 7, 2014): 559–71. http://dx.doi.org/10.1007/s10577-014-9445-5.

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Morris, Brett M., Suzanne L. Hawley, and Leslie Hebb. "Large Starspot Groups on HAT-P-11 in Activity Cycle 1." Research Notes of the AAS 2, no. 1 (February 6, 2018): 26. http://dx.doi.org/10.3847/2515-5172/aaac2e.

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Apak, Reşat, Mustafa Özyürek, Kubilay Güçlü, and Esra Çapanoğlu. "Antioxidant Activity/Capacity Measurement. 2. Hydrogen Atom Transfer (HAT)-Based, Mixed-Mode (Electron Transfer (ET)/HAT), and Lipid Peroxidation Assays." Journal of Agricultural and Food Chemistry 64, no. 5 (February 2016): 1028–45. http://dx.doi.org/10.1021/acs.jafc.5b04743.

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Wright, Diana, Torsten Wurm, Nicholas Polakowski, Jean-Michel Mesnard, and Isabelle Lemasson. "HBZ inhibits the HAT activity of the cellular coactivators p300 and CBP." Retrovirology 8, Suppl 1 (2011): A150. http://dx.doi.org/10.1186/1742-4690-8-s1-a150.

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Sarthi, Jessica, and Felice Elefant. "dTip60 HAT Activity Controls Synaptic Bouton Expansion at the Drosophila Neuromuscular Junction." PLoS ONE 6, no. 10 (October 27, 2011): e26202. http://dx.doi.org/10.1371/journal.pone.0026202.

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Yin, Bo-Kun, and Zhao-Qi Wang. "Beyond HAT Adaptor: TRRAP Liaisons with Sp1-Mediated Transcription." International Journal of Molecular Sciences 22, no. 22 (November 18, 2021): 12445. http://dx.doi.org/10.3390/ijms222212445.

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The members of the phosphatidylinositol 3-kinase-related kinase (PIKK) family play vital roles in multiple biological processes, including DNA damage response, metabolism, cell growth, mRNA decay, and transcription. TRRAP, as the only member lacking the enzymatic activity in this family, is an adaptor protein for several histone acetyltransferase (HAT) complexes and a scaffold protein for multiple transcription factors. TRRAP has been demonstrated to regulate various cellular functions in cell cycle progression, cell stemness maintenance and differentiation, as well as neural homeostasis. TRRAP is known to be an important orchestrator of many molecular machineries in gene transcription by modulating the activity of some key transcription factors, including E2F1, c-Myc, p53, and recently, Sp1. This review summarizes the biological and biochemical studies on the action mode of TRRAP together with the transcription factors, focusing on how TRRAP-HAT mediates the transactivation of Sp1-governing biological processes, including neurodegeneration.
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Dissertations / Theses on the topic "HAT activity"

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Dunphy, Elizabeth Louise. "TAF1 HAT activity in cell proliferation /." Thesis, Connect to this title online; UW restricted, 2003. http://hdl.handle.net/1773/6250.

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Hamed, Munerah. "Effect of p300 HAT Activity on Myogenic Differentiation." Thèse, Université d'Ottawa / University of Ottawa, 2013. http://hdl.handle.net/10393/23707.

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Skeletal muscle specification and differentiation programs are regulated by the myogenic regulatory factors which include Myf5, MyoD, myogenin and Mrf4. Upstream of the MRFs, the transcription co-activators and other intracellular and extracellular signals play crucial roles in regulating skeletal myogenesis. Histone acetyltransferase activity of p300 is required for Myf5 and MyoD expression. Furthermore, the MyoD core enhancer region is indispensable for MyoD expression. However, the mechanism by which p300 activates MyoD gene expression is to be determined. The histone acetyltransferase activity of p300 can be inhibited by small molecule inhibitors such as curcumin. Thus, using the inhibitor approach on stem cells is useful to investigate the role of p300 in activating MyoD expression during myogenesis. We here show that curcumin was able to inhibit stem cell determination and differentiation into skeletal myocytes. We also show that p300 is present, and histone acetylation is high at the core enhancer region. Therefore, we provide evidence that p300 is directly involved in MyoD gene expression during skeletal myogenesis.
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Brooks, Kimberlee. "Grade nine students and policy : perspectives of the school nutrition and physical activity environment." Thesis, Lethbridge, Alta. : University of Lethbridge, Faculty of Health Sciences, c2012, 2012. http://hdl.handle.net/10133/3114.

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The school environment is often the focus of healthy eating and physical activity policies intended to address childhood obesity. However, students are usually excluded from the development and implementation of such policies. The purpose of this study was to explore grade nine student perspectives regarding: supports and barriers to healthy eating and physical activity at school; strategies for improving the school nutrition and physical activity environments; and potential student contribution to the development, implementation, and evaluation of school-based policies. A qualitative approach utilizing focus groups and photovoice with 30 students from two schools in southern Alberta was used to collect data. Major themes included: access to healthy food choices; teacher influences; peer influences; access to physical activity opportunities; impact of marketing; and conflicting messages. Students can provide valuable insights to policy development, implementation and evaluation. Implications for future research and policy development are reviewed.
xi, 179 leaves : ill. ; 29 cm
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Husain, Ralf. "Hat die Belastung gestillter Kinder mit persistenten organischen Schadstoffen Einfluss auf natürliche Killerzellen?" Doctoral thesis, Humboldt-Universität zu Berlin, Medizinische Fakultät - Universitätsklinikum Charité, 2004. http://dx.doi.org/10.18452/15045.

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Gestillte Kinder sind über die Muttermilch mit persistenten organischen Schadstoffen (POPs) belastet. Tierexperimentelle Studien deuten auf eine besondere Empfindlichkeit des sich entwickelnden Immunsystems für POPs hin. Mögliche Effekte dieser Verbindungen auf den kindlichen Organismus sind bisher kaum untersucht. Es wurden potenzielle Einflüsse verschiedener POPs (PCDDs, PCDFs, PCBs, beta-HCH, HCB und pp-DDE) auf natürliche Killerzellen (NK-Zellen) und NK-Aktivität bei gestillten im Vergleich zu nicht gestillten Kindern untersucht. NK-Zellen sind eine Lymphozytensubpopulation (CD3-CD56/16+), die über ihre zytotoxische Aktivität allogene, virusinfizierte und maligne Zellen ohne vorherige Sensibilisierung töten kann. Es wurden 66 gesunde Kinder im Alter von 11-12 Monaten untersucht, davon waren 50 Kinder mindestens 4 Monate lang voll gestillt und 16 Kinder nicht gestillt. Aus einer Region mit bekannter erhöhter PCDD/PCDF-Belastung stammten 13 gestillte Kinder. Die NK-Zellzahlen wurden mittels Immunphänotypisierung am Durchflusszytometer bestimmt. Die Aktivität der NK-Zellen wurde mit einem nicht-radioaktiven, durchflusszytometrischen Zytotoxizitäts-Assay gemessen. Die POP-Konzentrationen im Blutfett der Probanden wurden kommerziell bestimmt. Weder bei den NK-Zellzahlen, noch bei der NK-Aktivität konnten zwischen den gestillten und nicht-gestillten Kindern signifikante Unterschiede im t-Test nachgewiesen werden. In Korrelationsanalysen zeigten sich keine signifikanten Einflüsse der POP-Konzentrationen auf NK-Zellzahlen und NK-Aktivität. Im Laufe der Untersuchung zeigte sich, dass der eingesetzte Zytotoxizitäts-Assay nur semiquantitative Daten lieferte. Die vorliegenden Ergebnisse und weitere Befunde bezüglich des Immunsystems der Probanden weisen darauf hin, dass die relativ hohe Belastung lange gestillter Säuglinge mit POPs nicht zu einer biologischen Wirkung im kindlichen Organismus führt. Angesichts der nachgewiesenen positiven Effekte des Stillens, kann diesbezüglich die bestehende Stillempfehlung bekräftigt werden.
Breast-fed infants are exposed to persistent organic pollutants (POPs) via breast milk. Animal studies indicate a special sensitivity of the maturing immune system to POPs. Possible effects of these compounds on the infantile organism are so far barely examined. Potential influences of several POPs (PCDDs, PCDFs, PCBs, beta-HCH, HCB and pp-DDE) on natural killer (NK) cells and NK activity of breast-fed infants in comparison to formula-fed infants were investigated. NK cells are a subset of lymphocytes (CD3-CD56/16+) that can kill allogeneic, virus-infected and malignant cells via their cytotoxic activity without prior sensitization. The study group consisted of 66 healthy infants examined at age 11 to 12 months, of which 50 infants were breast-fed and 16 infants were formula-fed. 13 breast-fed infants came from a region with known increased PCDD/PCDF-burden. Numbers of NK cells were measured by flow cytometric immunophenotyping. NK activity was analysed by a non-radioactive flow cytometric cytotoxicity assay. POP concentrations in the blood fat of the probands were calculated commercially. There were no significant differences between breast-fed and formula-fed infants concerning number and activity of NK cells in the t-test. Analysis of correlation showed no significant influences of POP concentrations on the number and activity of NK cells. In the course of the study the data obtained by the employed cytotoxicity assay proved to be only semiquantitative. The presented findings and further results concerning the immune system of the study subjects suggest that the relatively high burden of long-term breast-fed infants with POPs does not lead to a biological effect in the infantile organism. Regarding the proven positive effects of breast-feeding the existing recommendation to breastfeed can be encouraged.
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Worku, Netsanet, August Stich, Arwid Daugschies, Iris Wenzel, Randy Kurz, Rene Thieme, Susanne Kurz, and Gerd Birkenmeier. "Ethyl pyruvate emerges as a safe and fast acting agent against Trypanosoma brucei by targeting pyruvate kinase activity." Universitätsbibliothek Leipzig, 2015. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-179599.

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Background: Human African Trypanosomiasis (HAT) also called sleeping sickness is an infectious disease in humans caused by an extracellular protozoan parasite. The disease, if left untreated, results in 100% mortality. Currently available drugs are full of severe drawbacks and fail to escape the fast development of trypanosoma resistance. Due to similarities in cell metabolism between cancerous tumors and trypanosoma cells, some of the current registered drugs against HAT have also been tested in cancer chemotherapy. Here we demonstrate for the first time that the simple ester, ethyl pyruvate, comprises such properties. Results: The current study covers the efficacy and corresponding target evaluation of ethyl pyruvate on T. brucei cell lines using a combination of biochemical techniques including cell proliferation assays, enzyme kinetics, phasecontrast microscopic video imaging and ex vivo toxicity tests. We have shown that ethyl pyruvate effectively kills trypanosomes most probably by net ATP depletion through inhibition of pyruvate kinase (Ki = 3.0±0.29 mM). The potential of ethyl pyruvate as a trypanocidal compound is also strengthened by its fast acting property, killing cells within three hours post exposure. This has been demonstrated using video imaging of live cells as well as concentration and time dependency experiments. Most importantly, ethyl pyruvate produces minimal side effects in human red cells and is known to easily cross the blood-brain-barrier. This makes it a promising candidate for effective treatment of the two clinical stages of sleeping sickness. Trypanosome drug-resistance tests indicate irreversible cell death and a low incidence of resistance development under experimental conditions. Conclusion: Our results present ethyl pyruvate as a safe and fast acting trypanocidal compound and show that it inhibits the enzyme pyruvate kinase. Competitive inhibition of this enzyme was found to cause ATP depletion and cell death. Due to its ability to easily cross the bloodbrain- barrier, ethyl pyruvate could be considered as new candidate agent to treat the hemolymphatic as well as neurological stages of sleeping sickness.
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Hart, Jonathan Michael. "The influence of biofilm on the antifungal activity of amine oxide." View the abstract Download the full-text PDF version, 2009. http://etd.utmem.edu/ABSTRACTS/2009-015-Hart-index.htm.

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Thesis (M.S.)--University of Tennessee Health Science Center, 2009.
Title from title page screen (viewed on August 11, 2009). Research advisor: Jegdish P. Babu, Ph.D. Document formatted into pages (ix, 32 p. : ill.). Vita. Abstract. Includes bibliographical references (p. 28-31).
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Olsson, Kristina, and Helena Bäckström. "Tjugoåringars upplevelse av vilka faktorer som har haft betydelse för hur fysiskt aktiva de varit över tid." Thesis, Mälardalens högskola, Akademin för hälsa, vård och välfärd, 2016. http://urn.kb.se/resolve?urn=urn:nbn:se:mdh:diva-31502.

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SAMMANFATTNING Bakgrund: Fysisk aktivitet främjar hälsan och medför positiva effekter för kroppen. Barns och ungdomars fysiska aktivitet påverkas av olika faktorer på individ- och omgivningsnivå. Faktorerna interagerar med varandra och påverkar den fysiska aktiviteten på olika sätt och i varierande grad, faktorerna kan delas in i olika system i den Ekologiska systemteorin. Det kan exempelvis handla om föräldrars och vänners attityder och hur den fysiska miljön är utformad. Syfte: Syftet med denna studie var att utifrån ett Ekologiskt systemteoretiskt perspektiv undersöka 20-åringars upplevelse av vilka faktorer som har haft betydelse för hur fysiskt aktiva de har varit från 11-års ålder fram till idag. Metod: Studien genomfördes med en kvalitativ design med induktiv ansats. Fem deltagare intervjuades utifrån en intervjuguide innehållande semistrukturerade frågor. Kvalitativ innehållsanalys användes för att analysera och tolka materialet. Resultat: Analysen resulterade i 4 kategorier med 14 underkategorier. Kategorierna blev: ”Engagemang i idrotten påverkas av inre känslor och yttre krav”, ”Att vara en del av gemenskapen”, ”Den fysiska miljön och de socioekonomiska förutsättningarnas inverkan” och ”Andra möjliga påverkande faktorer på utövande”. Slutsats: Resultatet visade att de faktorer som har störst påverkan finns inom micro- och exosystemen. Faktorerna varierar med åldern och innefattar främst föräldrar, vänner och tillgänglighet.
ABSTRACT Background: Physical activity promotes health, and will have positive effects for the body. Children and adolescents physical activity is influenced by different factors at individual and environmental level. The factors integrate with each other and affect the physical activity in different ways and can be divided into different systems according to the ecological systems theory. It could for example involve the attitudes of parents and friends and how the physical environment is designed. Aim: Based on Ecological systems theory the purpose of this study is to examine 20-yearolds experience of which factors that have had an impact on their physical activity from 11 years old until today. Method: The study was conducted with a qualitative design with inductive approach. Five participants were interviewed based on an interview guide containing semi-structured questions. Qualitative content analysis was used to analyze and interpret the material. Result: The analysis resulted in 4 categories with 14 subcategories. The categories were: "Involvement in sport is affected by internal feelings and external demands", "Being a part of the community", "The impact of physical environment and socioeconomic conditions" and "Other possible factors affecting the activity". Conclusion: The result showed that the factors with the greatest impact is within the micro and exosystems. The factors varies over time and focuses on parents, friends and availability.
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Lloyd, Jonathan Andrew. "Hate Managers and Where They Target: An Analysis of Hate Crime as Hate Group Self-Help." Thesis, Virginia Tech, 2019. http://hdl.handle.net/10919/90883.

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I explore the relationships between hate group activity, community factors, and the likelihood of hate crime occurrence within a county area. I integrate considerations raised by Routine Activity and Social Control theorists as well as current hate crime literature to frame my concept of the hate manager, an agent of social control that utilizes hate crimes as a means of enacting extralegal self-help for hate groups. I explore the relationship between hate managers and hate crime by testing a model relating hate group activity and hate crime occurrences by location. Next, I correlate hate crime occurrences with hate group activity at the county level for the state of Virginia using public data. I find that a hate group's presence holds greater predictive power than nearly any other factor for hate crime likelihood. My findings illustrate the nature of hate crime as a means of social control; whereby hate groups act as a parochial order and maintain hierarchical relations between offenders and victims through means of disciplinary crimes. I conclude by outlining suggestions for future research into the role of the hate manager.
Master of Science
In my thesis, I ask the question of how hate groups methodically encourage where hate crimes occur. I do this by creating the concept of the hate manager. Hate managers are figures which influence would-be criminals into their illegal acts. They do this by stoking the fears necessary for them to act outside legal boundaries in reaction to some feeling of threat, an act known as self-help. Hate crimes, I argue, are a form of self-help where the feeling of threat is directed towards individuals belonging to some marginalized group. By looking at data collected by various agencies in the state of Virginia, I discover that the presence of a hate group in a county is a stronger predictor for such acts than any other factor for hate crime likelihood. By doing so, I demonstrate that hate crimes are a form of social control. That is, I argue that hate groups maintain a sense of order or ranking by means of illegal and disciplinary self-help in the form of hate crimes. I conclude my thesis by outlining suggestions for future exploration of the hate manager’s role.
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Aymard, Chloé. "Nouvelles méthodes électrochimiques pour le criblage d’inhibiteurs de transcétolases." Thesis, Lyon, 2018. http://www.theses.fr/2018LYSE1212/document.

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Cette thèse a pour objectif de développer une méthode électrochimique permettant de cribler à haut débit des inhibiteurs d'enzymes. Pour cela, une enzyme cible a été sélectionnée pour son intérêt thérapeutique, la transcétolase (TK) : elle est impliquée dans de nombreuses pathologies (cancer, maladies neurodégénératives, diabète…) et dans la survie de certains parasites pathogènes. Afin de mesurer l'activité de la TK, deux systèmes rapporteurs ont été développés, à l'aide de plaques de criblage électrochimique (PCE) constituées de 96 électrodes indépendantes. Le premier système rapporteur repose sur un système bienzymatique nécessitant l'intervention d'une enzyme auxiliaire, la galactose oxydase (GAOx) sous forme libre ou immobilisée dans la laponite. Cette enzyme est capable d'oxyder les produits de réaction de la TK et de produire du peroxyde d'hydrogène. Le format 96 des PCE a permis d'optimiser rapidement la détection électrochimique par ampérométrie pulsée par intermittence (IPA) d'activité oxydase et seulement 10 minutes sont nécessaires pour réaliser 96 mesures simultanées. Parallèlement, afin d'exploiter au maximum le système à 96 électrodes, cette détection d'activité oxydase a également été réalisée, en 10 minutes par électrochimiluminescence. Cette méthode offre l'avantage d'être plus sensible et moins variable que par IPA, mais limite l'utilisation de matrice d'immobilisation d'enzymes. La GAOx a ensuite été utilisée pour mesurer l'activité TK, cependant, les conditions réactionnelles n'étant pas optimales en présence du système bienzymatique (TK-GAOx), des temps d'incubation longs sont nécessaires et sont peu adaptées au criblage d'inhibiteurs de la TK. Un second système rapporteur ne nécessitant plus d'enzyme auxiliaire et faisant intervenir uniquement un seul substrat de la TK a été optimisé. Ce système repose sur l'oxydation de l'intermédiaire réactionnel formé par la fixation du cofacteur (la thiamine pyrophosphate) et du substrat sur l'enzyme par le ferricyanure. Cette méthode permet de mesurer 96 activités TK en seulement 7 minutes et a aisément été utilisée pour cribler une bibliothèque de molécules. Le criblage de 1360 molécules a permis d'identifier un nouvel inhibiteur de cet enzyme, présentant une CI50 de 63 μM. Le système électrochimique a également été utilisé pour déterminer le mécanisme d'inhibition (non compétitive pure partielle) et la constante d'inhibition associée (3,4 μM). Ces résultats sont inédits dans le domaine de l'électrochimie et offrent un large éventail d'applications que ce soit pour le criblage d'activité enzymatiques ou d'inhibiteurs d'enzymes
This thesis focuses on the development of a new electrochemical method allowing the high throughputscreening of enzyme inhibitors. For this purpose, a target enzyme has been selected for its therapeutic interest,transketolase (TK): this enzyme is involved in many diseases (cancer, neurodegenerative diseases, diabetes ...) and inthe survival of pathogenic parasites. To measure TK activity, two reporter systems have been developed by usingelectrochemical plates, composed of 96 independent electrodes.The first one is based on a bienzymatic system, requiring the use of an auxiliary enzyme, galactose oxidase(GAOx), in its soluble form or immobilized in laponite. This enzyme is able to oxidize TK products and producehydrogen peroxide. The 96-well format allowed to quickly optimize the electrochemical detection of oxidase activityby intermittent pulse amperometry (IPA) of oxidase activity and only 10 minutes are required to perform 96simultaneous measurements. In parallel, in order to harness the 96-well electrochemical system, this detection ofoxidase activity was also carried out in 10 minutes using electrochemiluminescence. This method is more sensitiveand less variable than IPA but is limited to the use of soluble enzymes. However, the reaction conditions are notoptimal for the bienzymatic system (TK-GAOx): long incubation times are required and are poorly adapted for thescreening of TK inhibitors.A second reporter system no longer requiring an auxiliary enzyme and involving only one TK substrate hasbeen optimized. This system relies on the oxidation by ferricyanide of the reactional intermediate resulted of thebinding of the cofactor (thiamine pyrophosphate) and the substrate. This method allows to measure 96 TK activitiesin only 7 minutes and was easily used to screen an in-house chemical library. The screening of 1360 molecules leadto the identification of a new TK inhibitor with an IC50 of 63 μM. This electrochemical system was also used todetermine the mechanism of inhibition (partial non-competitive mechanism) and the associated inhibition constant(3.4 μM). These results are innovative in the field of electrochemistry and offer a wide range of applications forenzymatic activity screening or the screening of enzymes inhibitors
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Hall, Lori L. "Race and Online Hate: Exploring the Relationship between Race and the Likelihood of Exposure to Hate Material Online." Diss., Virginia Tech, 2018. http://hdl.handle.net/10919/82037.

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This research examines the relationship between race and exposure to online hate material. The utilization of websites, weblogs, newsgroups, online games, radio broadcasts, online newsletters and a myriad of other online platforms has proliferated race-based hate groups in the US (Shafer 2002). According to the Southern Poverty Law Center (SPLC), the number of hate groups has been on the rise since the 1990s and continues to gain momentum with the advent of social media (Potok 2017). Exposure to separatist ideologies has propelled these radical rightwing groups into the mainstream by way of social media platforms, as they are "the most active producers of online hate material" (Costello, Hawdon, Ratliff, and Grantham 2016: pg. 313). That dissemination of radical rightwing ideologies, such as white supremacy, racial purity and racial solidarity, exists is not enough in understanding what individuals are exposed to race-based hate ideologies in online platforms. Exposure is the key to understanding the growth of these race-based hate groups and ways of countering the efforts to disseminate radical rightwing ideologies due to its relationship to hate group emergence and persistence. More so, understanding how these groups target individuals and recruit through social networking sites can provide insight into exposure. Exposure to hate material aids groups in recruiting new members and victimizing potential targets. In the same manner, exposure to hate material is victimization of those who are exposed. In a sample collected by Costello et al. (2016a), of those exposed to hate material online nearly half centered on race. Thus, it is tantamount that research be conducted examining the role that race plays in determining who is exposed to hate material online, and how individuals react to hate material based on race. This dissertation will examine the importance of exposure to hate. Specifically, this dissertation will analyze survey data gathered from the Online Extremism Survey using logistic regression analysis and linear regression to understand exposure to hate material online and routine activity theory.
Ph. D.
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Books on the topic "HAT activity"

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1968: Das Jahr, das alles verändert hat. München: Piper, 1998.

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Ted, Enik, ed. Dr. Seuss' cat in the hat, the movie!: Wipe-off talking activity book. Lincolnwood, Ill: Publications International, 2003.

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"Der Hass hat uns geeint": Junge Rechtsextreme und ihr Ausstieg aus der Szene. Frankfurt/Main: Campus, 2006.

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Wer nicht kämpft, hat schon verloren: Frauen und der Mut zur Macht. Reinbek bei Hamburg: Rowohlt, 1986.

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Min zhu nü shen hao hang hai ri ji. Taibei Shi: Lian he bao she, 1990.

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Liang Han Wei Jin Nan Bei chao shi qi Guandong hao zu yan jiu. Chengdu Shi: Sichuan da xue chu ban she, 2007.

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Sarang to myŏngye to irŭm to namgim ŏpsi: Na ŭi han salmae. Sŏul-si: Hanʼgyŏre Chʻulpʻan, 2009.

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Hai wai Hua qiao yu xin hai ge ming. Guangzhou: Ji nan da xue chu ban she, 2011.

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Armiwulan, Hesti. Menggugat hak politik perempuan! [Jakarta]: Kerjasama Konsorsium Swara Perempuan (KSP) dan the Ford Foundation, 2005.

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Christiansen, Peter Munk. De som meget har--: Store danske virksomheder som politiske aktører. Århus, Danmark: Magtudredningen, 2003.

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Book chapters on the topic "HAT activity"

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Janes, Robert R., and Richard Sandell. "Posterity has arrived." In Museum Activism, 1–22. Abingdon, Oxon ; New York, NY : Routledge, 2019. | Series: Museum meanings: Routledge, 2019. http://dx.doi.org/10.4324/9781351251044-1.

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Gerson, Joseph. "He Has Never Stopped." In Chomsky for Activists, 200–203. New York, NY: Routledge, 2021. | Series: Universalizing resistance: Routledge, 2020. http://dx.doi.org/10.4324/9781003105619-30.

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Asea, Alexzander, and Punit Kaur. "The Chaperokine Activity of HSPA1A." In Heat Shock Proteins, 201–13. Dordrecht: Springer Netherlands, 2012. http://dx.doi.org/10.1007/978-94-007-4740-1_13.

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Nagai, Katsuya, Akiko Hashida, and Hachiro Nakagawa. "Dimethylarginine has Digitalis-Like Activity." In Guanidines 2, 115–22. Boston, MA: Springer US, 1989. http://dx.doi.org/10.1007/978-1-4613-0821-8_14.

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Kaur, Punit, and Alexzander A. A. Asea. "The Chaperokine Activity of Heat Shock Proteins." In Heat Shock Proteins, 3–22. Cham: Springer International Publishing, 2019. http://dx.doi.org/10.1007/978-3-030-02254-9_1.

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Ronczkowski, Michael R. "Behavioral Traits and Suspicious Activity." In Terrorism and Organized Hate Crime, 61–82. Fourth edition. | Boca Raton : CRC Press, [2018]: CRC Press, 2017. http://dx.doi.org/10.4324/9781315203133-4.

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Mande, Shekhar C., C. M. Santosh Kumar, and Aditi Sharma. "Evolution of Bacterial Chaperonin 60 Paralogues and Moonlighting Activity." In Heat Shock Proteins, 101–21. Dordrecht: Springer Netherlands, 2013. http://dx.doi.org/10.1007/978-94-007-6787-4_7.

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Haegele, Justin A., and Wesley J. Wilson. "10 Things I Hate about ‘Inclusion' in Physical Education." In Reflexivity and Change in Adaptive Physical Activity, 21–33. London: Routledge, 2022. http://dx.doi.org/10.4324/9781003196747-4.

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Belval, Luke N., and Lawrence E. Armstrong. "Comparative Physiology of Thermoregulation." In Sport and Physical Activity in the Heat, 3–14. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-70217-9_1.

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Huggins, Robert A., William M. Adams, Mark J. Buller, and David P. Looney. "Technology." In Sport and Physical Activity in the Heat, 163–89. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-70217-9_10.

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Conference papers on the topic "HAT activity"

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Yan, Jie, Xianlin Zeng, Anfu Zhou, and Huadong Ma. "MM-HAT: Transformer for Millimeter-Wave Sensing Based Human Activity Recognition." In GLOBECOM 2022 - 2022 IEEE Global Communications Conference. IEEE, 2022. http://dx.doi.org/10.1109/globecom48099.2022.10000673.

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Milanović, Žiko, Edina Avdović, Marko Antonijević, and Zoran Marković. "DIRECT SCAVENGING ACTIVITY OF 4,7-DIHYDROXYCOUMARIN DERIVATIVE TOWARDS SERIES OF CHLOROMETHYLPEROXY RADICALS." In 1st INTERNATIONAL Conference on Chemo and BioInformatics. Institute for Information Technologies, University of Kragujevac,, 2021. http://dx.doi.org/10.46793/iccbi21.462m.

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In this study, the mechanisms of antiradical activity of the previously synthesized 4,7- dihydroxycoumarin derivative:(E)-3-(1-((4-hydroxyphenyl)amino)ethylidene)-2,4- dioxochroman-7-yl acetate (A-4OH) according to the series of chloromethylperoxy radicals: OOCH2Cl (chloromethylperoxy), OOCHCl2 (dichloromethylperoxy) and OOCCl3 (trichloromethyl peroxy) were investigated. To assess the mechanisms of antiradical activity of A-4OH against chloromethylperoxy radicals, the following mechanistic pathways were examined: Hydrogen Atom Transfer (HAT), Single-Electron Transfer followed by Proton Transfer (SET-PT), and Radical Adduct Formation (RAF). The investigations were performed in water using sophisticated computational methods. The results of the study show that the activity against selected radicals decreases in series •OOCCl3 > •OOCHCl2 > •OOCH2Cl. The results of thermodynamic and kinetic parameters indicate that A-4OH inactivates selected radicals via the HAT mechanism.
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Cvijetić, Ilija, Miljan Bigović, Petar Ristivojević, Maja Vitorović-Todorović, Mire Zloh, and Dušanka Milojković-Opsenica. "THERMODYNAMICS OF THE ANTIOXIDANT ACTIVITY OF HUMULONES AND OTHER ANTIOXIDANTS FROM BEER – A MOLECULAR MODELING APPROACH." In 1st INTERNATIONAL Conference on Chemo and BioInformatics. Institute for Information Technologies, University of Kragujevac, 2021. http://dx.doi.org/10.46793/iccbi21.408c.

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Recent experimental study identified eight potent antioxidants in German beers, including isoxanthohumol, (R)- and (S)-adhumulone, cis– and trans-iso-adhumulone, cis– and trans-iso- n-humulone, and desdimetyhyl-octahydro-iso-cohumulone. To provide insights into the structural basis of their radical scavenging activity, we calculated the thermodynamic feasibility of two common antioxidant mechanisms, hydrogen atom transfer (HAT) and single electron transfer followed by proton transfer (SET-PT), using the density functional theory (DFT) with B3LYP/6-311g++(2d,2p) method in the gas phase and implicit solvation model of water. The calculated bond dissociation enthalpies (BDEs) and ionization potential (IP) of all compounds were compared with the corresponding values for resveratrol, a highly potent antioxidant found in red wine. The fully reduced humulone isomer, desdimetyhyl-octahydro-iso-cohumulone, could scavenge free radicals via HAT as revealed by BDEs 5.1 and 23.9 kJ/mol lower than the values for resveratrol in gas phase and water, respectively. Furthermore, the enolic –OH group was identified as the pharmacophoric hotspot for the interaction of humulones with the reactive free radicals. The HAT potency of this group is significantly reduced through the formation of strong intramolecular hydrogen bond (IHB) with the β-keto group. Moreover, the SET-PT mechanism was thermodynamically favorable for isoxanthohumol. These results strongly suggest higher antioxidant activity of beers with the increased content of the reduced forms of humulones and their isomers.
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Antonijević, Marko, Žiko Milanović, Dušica Simijonović, Zoran Marković, and Snežana Marković S. Zoran. "ANTIOXIDATIVE POTENCY AND RADICAL SCAVENGING ACTIVITY OF SELECTED COUMARIN-HYBRIDS." In 1st INTERNATIONAL Conference on Chemo and BioInformatics. Institute for Information Technologies, University of Kragujevac,, 2021. http://dx.doi.org/10.46793/iccbi21.458a.

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In previous studies, it was found that coumarins with hydrazide moiety show good antioxidative potential, while similar coumarins with hydrazone moiety are good anticancer agents. In this paper, the antioxidative potency and radical scavenging activity of two coumarin hydrazone derivatives were investigated. For this purpose, density functional theory method M062X with 6-311G++(d,p) basis set was implemented. It was found that investigated compounds exhibit good antioxidative potency, with very similar BDE values regardless of the position involved. On the other hand, PA values show that a preferable functional group for proton loss depends on the position of the OH group. In ortho position, OH group shows lower antioxidative potency than NH group, while in the same time in para position OH group is favourable position for antioxidative activity reactions. A similar situation is obtained by investigation of radical scavenging mechanisms, with the more pronounced difference in BDE between the positions in favour of the NH group. While SPLET is the most probable mechanism which is in competition with HAT in some cases (hydroxy radical) SET-PT was found to be a non-operative mechanistic pathway.
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Marković, Svetlana, and Jelena Tošović. "CHLOROGENIC ACID – APPLICATION OF CONTEMPORARY DENSITY FUNCTIONALS TO A SINGLE MOLECULE." In 1st INTERNATIONAL Conference on Chemo and BioInformatics. Institute for Information Technologies, University of Kragujevac,, 2021. http://dx.doi.org/10.46793/iccbi21.081m.

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Although chlorogenic acid (5-O-caffeoylquinic acid, 5CQA) is a dietary phenol known for its pharmacological and nutritional properties, its structural features and mechanisms of oxidative action have not been completely elucidated. Clarification of the 5CQA structure was conducted by comparing the experimental and simulated IR, Raman, 1H-NMR, 13C-NMR, and UV spectra. For this purpose, a comprehensive conformational analysis of 5CQA was performed to reveal its most stable conformations in the gas-state and solution. Excellent agreement between all experimental and simulated spectra indicates correct arrangement of the atoms in the 5CQA molecule. In addition, the most stable conformation in solution coincides with that predicted with sophisticated NMR experiments. The quantum mechanics–based test for overall free-radical scavenging activity was applied for the investigation of antioxidative capacity of 5CQA relative to trolox (6-hydroxy-2,5,7,8- tetramethylchroman-2-carboxylic acid, Tx) as a reference compound. Hydrogen atom transfer (HAT), radical adduct formation (RAF), sequential proton loss electron transfer (SPLET), and single electron transfer – proton transfer (SET-PT) reactions of 5CQA and Tx with HO· and CH3OO· radicals were examined in benzene, pentyl ethanoate, and basic aqueous solutions. In non-polar solvents 5CQA reacts with HO· via HAT and RAF mechanisms, whereas HAT is the only reaction pathway with CH3OO·. At physiological conditions 5CQA exists in the form of monoanion and dianion. Both anionic forms undergo only HAT mechanism with CH3OO·. With HO·, the anions conform to the HAT, RAF, SPLET, and SET-PT mechanisms. Because all reactions of dianion are diffusion controlled, its contribution to scavenging HO· is comparable to that of more abundant monoanion. The calculated rate constant for overall reaction of 5CQA with HO· is in perfect agreement with the corresponding experimental value. The order of reactivity toward selected free radicals is the same in nonpolar and polar solutions: in comparison to Tx, 5CQA is more reactive toward HO·, but less reactive toward CH3OO·. Very good agreement between the experimental and calculated results confirms the ability of contemporary density functionals to quantify subtle physico-chemical interactions.
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Milanovic, Žiko, Ana Kesic, Edina Avdovic, Jelena Đorovic Jovanovic, and Dejan Milenkovic. "UTICAJ pH VREDNOSTI NA ANTIRADIKALSKI KAPACITET 4,7- DIHIDROKSIKUMARINA." In XXVI savetovanje o biotehnologiji sa međunarodnim učešćem. University of Kragujevac, Faculty of Agronomy, 2021. http://dx.doi.org/10.46793/sbt26.481m.

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The mechanisms of antiradical activity of 4,7-dihydroxycoumarin against peroxy (HOO•) radical were investigated. For this purpose were used sophisticated computational methods. Investigation was performed in the water, at different pH values (0-14). Acid-base species, which are represented in diverse molar fraction (f) at different pH values, contribute to a comprehensive examination of antiradical capacity. An overall rate constant (koverall) of the favorable reaction pathways (HAT and RAF) of 2.07×102 M-1s-1 at physiological pH indicates a moderate ability to neutralization HOO• radicals. A value of 1.05×103 M-1 s-1 occurring at pH > 10 indicates a maximum antiradical capacity of 4,7-dihydroxymumarin against the HOO• radical.
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Liote, F., M. P. Wautier, E. Savariau, H. Setiadi, and J. L. Wautier. "INHIBITION OF ENDOTHELIAL CELL PROLIFERATION BY NORMAL HUMAN MONOCYTES." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643170.

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Human peripheral blood monocytes and macrophages possess factors which are capable of inhibiting or stimulating endothelial cell proliferation. We have further explored if such activity is due to cytotoxic effects of monocytes. Normal mononuclear cells were isolated first by density gradient. Monocytes were then purified by three different techniques: 1) counter centrifugation elutriation (CCE) (Beckman) 2) selective adhesion to gelatin-plasma (GPI) 3) selective adhesion to fibronectin (Fn). Cytotoxicity was estimated by counting the release of 51cr used to label the human umbilical vein endothelial cells (HUVE) prior to the addition of monocytes. Whilst [3H] thymidine incorporation by HUVE permitted us to measure the effect of monocytes on the growth of the endothelial cells. Monocytes were incubated with HUVE (12×103) for 24 to 36h at various concentrations '(1.5-12×103). No cytotoxic effect could be demonstrated but an inhibition of [3h] thymidine uptake was observed and was dependent upon monocytes concentration. Monocytes isolated on GP1 exhibited a significantly higher inhibitory effect (p<0.05) compared to those purified on Fn or by CCE.(GP1: 85±6%, Fn:58±6%, CCE:67±5%). These results indicated t*hat normal monocytes can inhibit endothelial cell proliferation. This activity appeared to be higher when monocytes were isolated on GP1 which suggest that the adhesion on this surface could stimulate monocytes not only by its fibronectin receptor. This inhibitory activity of monocyte on endothelial cells proliferation could be different in patients with vascular disorders.
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"Nootropic activity of humic substances." In Seventh International Conference on Humic Innovative Technologies "Humic substances and technologies for resilience" (HIT – 2022). NP CBR "Humus Sapiens", 2022. http://dx.doi.org/10.36291/hit.2022.065.

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"The reasons of humic substances biological activity." In Seventh International Conference on Humic Innovative Technologies "Humic substances and technologies for resilience" (HIT – 2022). NP CBR "Humus Sapiens", 2022. http://dx.doi.org/10.36291/hit.2022.088.

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"Laurus nobilis Has Antibacterial Activity Against Staphylococcus aureus." In International Conference on Food, Biological and Medical Sciences. International Institute of Chemical, Biological & Environmental Engineering, 2014. http://dx.doi.org/10.15242/iicbe.c0114594.

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Reports on the topic "HAT activity"

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Kerem, Zohar, Yael Vodovotz, David Bonfil, Steven J. Schwartz, and Mark Failla. Do Saponins Present in Model Systems and Legume Bread Modulate Cholesterol Absorption in vitro and in vivo? United States Department of Agriculture, August 2011. http://dx.doi.org/10.32747/2011.7592656.bard.

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No abstract. The overall general goal of the research project was to provide evidence for the beneficial biological activity of saponins from edible legumes, and to incorporate them into bread hat will become a functional food. Its functionality is in its ability to attenuate the absorption of dietary lipids and cholesterol.
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Patton, Brenda, Sara Cutler, James S. Dickson, Steven M. Lonergan, and Chad H. Stahl. Colicin E1 has High Antimicrobial Activity against Listeria Monocytogenes in Culture and on Packaged Ham. Ames (Iowa): Iowa State University, January 2007. http://dx.doi.org/10.31274/ans_air-180814-79.

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Nasol, D. Heat of Hydration of Low Activity Cementitious Waste Forms. Office of Scientific and Technical Information (OSTI), July 2015. http://dx.doi.org/10.2172/1204364.

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Second Physical Activity Almanac. Chair Andrea Ramírez Varela. Ediciones Uniandes, November 2021. http://dx.doi.org/10.51572/202102.

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En este almanaque, GoPA! presenta el Segundo Juego completo de 164 Tarjetas de País con la evidencia disponible hasta 2019-2020 sobre investigación, políticas y vigilancia de la actividad física en todo el mundo, correspondiente a una cobertura global del 75,6%. El proyecto ha reunido con éxito a representantes de casi todos los países del mundo para abordar la actividad física como un problema de salud pública. En 2015, se lanzaron 139 Country Cards y en 2020 este número ha aumentado a 164 Country Cards. El propósito del Almanaque es ayudar a los países a determinar sus necesidades e iniciar o mejorar la recopilación de datos estandarizados, los sistemas de vigilancia, la formulación de políticas, la investigación, las estadísticas de carga de salud, el desarrollo de programas y la evaluación relacionada con la actividad física. Alentamos a los gobiernos, investigadores, defensores de la actividad física y al público en general a utilizar este recurso para mejorar los niveles de actividad física de la población.
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McCoskey, Jacob K. Instrument Qualification of Custom Fabricated Water Activity Meter for Hot Cell Use. Office of Scientific and Technical Information (OSTI), January 2014. http://dx.doi.org/10.2172/1119690.

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Niu, Gang. Imaging Heat Shock Protein 90 (Hsp90) Activity in Hormone-Refractory Prostate Cancer. Fort Belvoir, VA: Defense Technical Information Center, March 2009. http://dx.doi.org/10.21236/ada506361.

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Hodges, Thomas K., and David Gidoni. Regulated Expression of Yeast FLP Recombinase in Plant Cells. United States Department of Agriculture, September 2000. http://dx.doi.org/10.32747/2000.7574341.bard.

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Research activities in both our laboratories were directed toward development of control of the FLP/frt recombination system for plants. As described in the text of the research proposal, the US lab has been engaged in developing regulatory strategies such as tissue-specific promoters and the steroid-inducible activation of the FLP enzyme while the main research activities in Israel have been directed toward the development and testing of a copper-regulated expression of flp recombinase in tobacco (this is an example of a promoter activation by metal ions). The Israeli lab hat additionally completed experiments of previous studies regarding factors affecting the efficiency of recombinase activity using both a gain-of-function assay (excisional-activation of a gusA marker) and loss of function assay (excision of a rolC marker) in tobacco. Site-specific recombinase systems, in particular the FLP/frt and R/RS systems of yeast and the Cre/lox system of bacteriophage P1, have become an essential component of targeted genetic transformation procedures both in animal and plant organisms. To provide more flexibility in transgene excisions by the recombinase systems as well as gene targeting, and to widen possible applications, the development of controlled or regulated recombination systems is highly desirable and was therefore the subject of this research proposal. There are a few possible mechanisms to regulate expression of a recombinase system. They include: 1) control of the recombination system by having the target sites (e.g. frt) in one plant and the flp recombinase gene in another, and bringing the two together by cross fertilization. 2) regulation of promoter activities by external stimuli such as temperature, chemicals, metal ions, etc. 3) regulation of promoter activities by internal signals, i.e. cell- or tissue-specific, or developmental regulation. 4) regulation of enzyme activity by providing cofactors essential for biochemical reactions to take place such as steroid molecules in conjunction with a steroid ligand-binding protein (domains). During the course of this research our major emphasis have been focused toward studying the feasibility of hybrid seed production in Arabidopsis, using FLP/frt. Male-sterility was induced using the antisence of a pollen- and tapetum-specific gene, bcp1, isolated from Arabidopsis. The sterility inducing gene was flanked by frt sites. Upon cross pollination of flowers of male-sterile plants with pollen from FLP-containing plants, viable seeds were produced, and the progeny hybrid plants developed normally. The major achievement from this work is the first demonstration of using a site-specific recombinase to restore fertility in male-sterile plants (see attached paper, Luo et al., Plant J 2000; 23:423-430). The implication from this finding is that site-specific recombination systems can be applied in crop plants as a useful alternative method for hybrid seed production.
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McIntyre, Phillip, Susan Kerrigan, and Marion McCutcheon. Australian Cultural and Creative Activity: A Population and Hotspot Analysis: Albury-Wodonga. Queensland University of Technology, 2020. http://dx.doi.org/10.5204/rep.eprints.206966.

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Albury-Wodonga, situated in Wiradjuri country, sits astride the Murray River and has benefitted in many ways from its almost equidistance from Sydney and Melbourne. It has found strength in the earlier push for decentralisation begun in early 1970s. A number of State and Federal agencies have ensured middle class professionals now call this region home. Light industry is a feature of Wodonga while Albury maintains the traditions and culture of its former life as part of the agricultural squattocracy. Both Local Councils are keen to work cooperatively to ensure the region is an attractive place to live signing an historical partnership agreement. The region’s road, rail, increasing air links and now digital infrastructure, keep it closely connected to events elsewhere. At the same time its distance from the metropolitan centres has meant it has had to ensure that its creative and cultural life has been taken into its own hands. The establishment of the sophisticated Murray Art Museum Albury (MAMA) as well as the presence of the LibraryMuseum, Hothouse Theatre, Fruit Fly Circus, The Cube, Arts Space and the development of Gateway Island on the Murray River as a cultural hub, as well as the high profile activities of its energetic, entrepreneurial and internationally savvy locals running many small businesses, events and festivals, ensures Albury Wodonga has a creative heart to add to its rural and regional activities.
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Gibbs, A., and R. S. Thomason. Compilation of Data on Radionuclide Data for Specific Activity, Specific Heat and Fission Product Yields. Office of Scientific and Technical Information (OSTI), September 2000. http://dx.doi.org/10.2172/762019.

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Locy, Robert D., Hillel Fromm, Joe H. Cherry, and Narendra K. Singh. Regulation of Arabidopsis Glutamate Decarboxylase in Response to Heat Stress: Modulation of Enzyme Activity and Gene Expression. United States Department of Agriculture, January 2001. http://dx.doi.org/10.32747/2001.7575288.bard.

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Most plants accumulate the nonprotein amino acid, g-aminobutyric acid (GABA), in response to heat stress. GABA is made from glutamate in a reaction catalyzed by glutamate decarboxylase (GAD), an enzyme that has been shown by the Israeli PI to be a calmodulin (CaM) binding protein whose activity is regulated in vitro by calcium and CaM. In Arabidopsis there are at least 5 GAD genes, two isoforms of GAD, GAD1 and GAD2, are known to be expressed, both of which appear to be calmodulin-binding proteins. The role of GABA accumulation in stress tolerance remains unclear, and thus the objectives of the proposed work are intended to clarify the possible roles of GABA in stress tolerance by studying the factors which regulate the activity of GAD in vivo. Our intent was to demonstrate the factors that mediate the expression of GAD activity by analyzing the promoters of the GAD1 and GAD2 genes, to determine the role of stress induced calcium signaling in the regulation of GAD activity, to investigate the role of phosphorylation of the CaM-binding domain in the regulation of GAD activity, and to investigate whether ABA signaling could be involved in GAD regulation via the following set of original Project Objectives: 1. Construction of chimeric GAD1 and GAD2 promoter/reporter gene fusions and their utilization for determining cell-specific expression of GAD genes in Arabidopsis. 2. Utilizing transgenic plants harboring chimeric GAD1 promoter-luciferase constructs for isolating mutants in genes controlling GAD1 gene activation in response to heat shock. 3. Assess the role of Ca2+/CaM in the regulation of GAD activity in vivo in Arabidopsis. 4. Study the possible phosphorylation of GAD as a means of regulation of GAD activity. 5. Utilize ABA mutants of Arabidopsis to assess the involvement of this phytohormone in GAD activation by stress stimuli. The major conclusions of Objective 1 was that GAD1 was strongly expressed in the elongating region of the root, while GAD2 was mainly expressed along the phloem in both roots and shoots. In addition, GAD activity was found not to be transcriptionally regulated in response to heat stress. Subsequently, The Israeli side obtained a GAD1 knockout mutation, and in light of the objective 1 results it was determined that characterization of this knockout mutation would contribute more to the project than the proposed Objective 2. The major conclusion of Objective 3 is that heat-stress-induced changes in GAD activity can be explained by heat-stress-induced changes in cytosolic calcium levels. No evidence that GAD activity was transcriptionally or translationally regulated or that protein phosphorylation was involved in GAD regulation (objective 4) was obtained. Previously published data by others showing that in wheat roots ABA regulated GABA accumulation proved not to be the case in Arabidopsis (Objective 5). Consequently, we put the remaining effort in the project into the selection of mutants related to temperature adaptation and GABA utilization and attempting to characterize events resulting from GABA accumulation. A set of 3 heat sensitive mutants that appear to have GABA related mutations have been isolated and partially characterized, and a study linking GABA accumulation to growth stimulation and altered nitrate assimilation were conducted. By providing a better understanding of how GAD activity was and was not regulated in vivo, we have ruled out the use of certain genes for genetically engineering thermotolerance, and suggested other areas of endeavor related to the thrust of the project that may be more likely approaches to genetically engineering thermotolerance.
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