William Taylor - US grants

A recently isolated maize transposable element, Mul, has been used to generate new mutants. We propose to use rapid screening methods to recover mutants deficient in one or more photosynthetic activities or groups of functionally related proteins. Further characterization of selected mutants will allow us to concentrate on those whose gene products appear to regulate the synthesis or assembly of photosynthetic complexes. Since the mutants were caused by Mul insertions, a Mul probe will be used to isolate mutants by molecular cloning. The role of the cloned genes in synthesis or assembly of multiprotein photosynthetic complexes will be determined.

The levels of the 5S RNA gene specific positive transcription factor, TFIIIA, appear to play a critical role in the developmental regulation of 5S RNA genes in Xenopus. The expression of the TFIIIA gene is known to be developmentally regulated at the level of mRNA accumulation. It is the objective of this proposal to identify the factors which control TFIIIA gene expression in oocytes and in somatic cells. This work is specifically directed at examining the molecular interactions involved in expression of-the TFIIIA gene in oocytes and somatic cells. In addition, differences between the expression of the TFIIIA gene in oocytes and somatic cells will be examined. Experiments to define cis-acting sequences of the TFIIIA gene and surrounding genomic DNA sequences, which affect the transcription of the gene, will be performed using standard gene fusion and transfection techniques. Deletion mutations of the TFIIIA gene and surrounding genomic DNA sequences will be prepared by in vitro mutagenesis protocols. The mutated sequences will then be assayed for the ability to direct specific transcription of the TFIIIA gene in oocytes and somatic cells. In this manner, it should be possible to define DNA sequences from the TFIIIA gene and surrounding genomic regions which act in a positive and/or negative fashion to control the expression of the TFIIIA gene in oocytes and somatic cells. Differences in the expression of the deletion mutants in oocytes and somatic cells will be examined carefully. These differences are likely to define regions of the TFIIIA gene and surrounding genomic sequences which are important in the tissue specificity of the TFIIIA gene. Finally, the interaction of cis-acting sequences which affect the expression of the TFIIIA gene with proteins (trans-acting factors) from oocytes and somatic cells will be examined using various DNA- protein binding assays. The presence or absence in oocytes or somatic cells or cell type specific modification of the DNA binding proteins may play a crucial role in the expression of the TFIIIA gene. Purification of the proteins which interact with cis-acting sequences which control the expression of the TFIIIA gene will be attempted. The purification of these proteins should allow the detailed examination of their role in the transcription of the TFIIIA gene in oocytes and somatic cells, including any modifications and tissue specificity.

The long term goal of this project is to find out which of the gamma-ray induced lesions in DNA are responsible for the biological effects; lethality, mutagenesis and chromosomal rearrangements. The specific aim of this proposal is to determine which role each kind of gamma-ray induced DNA-damage plays in causing lethality, mutagenesis and recombination in an E. coli- plasmid DNA system. The plasmid DNA will be irradiated in vitro under controlled conditions so that the amounts and ratios of the kinds of damage can be changed. The amounts of damage will be determined by measuring the conversion of the form I to forms II and III DNA under neutral conditions and after alkaline hydrolysis. The biological effects will be measured by transfecting the irradiated plasmid and the separated DNA forms into calcium treated E. coli host cells. The irradiated plasmid will be separated into forms I, II and III by agarose gel electrophoresis and sucrose gradient sedimentation. Lethality will be determined from the loss of tetracycline or ampicillin resistant colonies depending on which plasmid is used. Mutagenesis will be measured as forward mutations of the Lac gene in plasmid PBRPI which contains the E. coli Lac operon cloned into PBR322. The host cells in this case will be delta lac. Recombination will be measured by counting the number of Lac-colonies produced when this plasmid is transfected into an E. coli host containing a mutant (Lac Y1) allele. The recombinations will be confirmed by plasmid isolation and restriction enzyme analysis to locate the Pst I and Pvu II sites associated with the Lac Y1 mutation. The structure of the Lac genes in E. coli host DNA will be checked by Southern blotting of restriction fragments separated on agarose gels. Using particular combinations of Lac+ and Lac- hosts with plasmids pBRM4 and pBRM5, which contain subcloned segments of the Lac Y gene, the number of reciprocal and non-reciprocal recombinations from host-to-plasmid and plasmid-to-host will be determined. In the long term this system will be used to measure and/or predict the biological effects of radio-protective or sensitizing agents which may be used in radiation therapy or for protection of populations likely to be exposed to gamma-rays or other highly ionizing radiation.

DESCRIPTION (Adapted from the applicant's abstract): The primary goal of this work is to understand the molecular mechanism by which the Transcription Factor IIIA (TFIIIA) gene is developmentally regulated. The studies to date have concentrated on the identification of cis-elements required for the expression of the TFIIIA gene in immature oocytes, mature oocytes and somatic cells. Eight or more of these cis-elements have been identified, several of which appear to be developmentally regulated. The developmentally regulated elements are functionally expressed at different times, one is specific for immature oocytes and the others (several negative and one positive) are "embryo-specific" (not functional in oocytes). The times at which these factors are regulated correspond to times in development when TFIIIA mRNA levels change (10-fold more TFIIIA mRNA in immature oocytes than in mature oocytes and approximately 10-5 more TFIIIA mRNA in mature oocytes than in post-gastrula embryos). The investigator has identified trans-acting factors which bind to most of these elements and have obtained putative cDNA clones for one of the developmentally regulated factors (the one which binds to the immature oocyte-specific element). This proposal concentrates on expanding the results on the interactions between these elements and the further characterization of factors which bind to these elements. The investigator proposes to isolate the various trans-acting factors and/or isolate cDNA clones encoding these factors. These cDNA clones will then provide one with molecular probes for these factors. The functions of these factors and interactions between them and other transcription factors will then be examined using in vivo and in vitro approaches. Qualitative and quantitative differences in the structure, activity, and interactions of these factors will be determined at various times during development.

DESCRIPTION (Adapted from the applicant's abstract): The primary goal of this work is to understand the molecular mechanism by which the Transcription Factor IIIA (TFIIIA) gene is developmentally regulated. The studies to date have concentrated on the identification of cis-elements required for the expression of the TFIIIA gene in immature oocytes, mature oocytes and somatic cells. Eight or more of these cis-elements have been identified, several of which appear to be developmentally regulated. The developmentally regulated elements are functionally expressed at different times, one is specific for immature oocytes and the others (several negative and one positive) are "embryo-specific" (not functional in oocytes). The times at which these factors are regulated correspond to times in development when TFIIIA mRNA levels change (10-fold more TFIIIA mRNA in immature oocytes than in mature oocytes and approximately 10-5 more TFIIIA mRNA in mature oocytes than in post-gastrula embryos). The investigator has identified trans-acting factors which bind to most of these elements and have obtained putative cDNA clones for one of the developmentally regulated factors (the one which binds to the immature oocyte-specific element). This proposal concentrates on expanding the results on the interactions between these elements and the further characterization of factors which bind to these elements. The investigator proposes to isolate the various trans-acting factors and/or isolate cDNA clones encoding these factors. These cDNA clones will then provide one with molecular probes for these factors. The functions of these factors and interactions between them and other transcription factors will then be examined using in vivo and in vitro approaches. Qualitative and quantitative differences in the structure, activity, and interactions of these factors will be determined at various times during development.

0085095
Epstein
The purpose of this grant is to explore the possibilities of greatly
expaniding a successful outreach program developed at the Department of Physics and Astronomy at California State University - Los Angeles, known at the University Preparatory Program (UPP). This effort will include data gathering and making contact with potential partners in universities, high schools, and corporations in greater Los Angeles and the southwestern United States.

Professor William Taylor of the University of Central Arkansas is supported by the Experimental Physical Chemistry program to perform experimental studies on metal ions complexed with hydrocarbons in the gas phase. Transition metals are frequently used as catalysts for activating hydrocarbons, but it is not understood how reactivity depends on the electronic state of the metal. This proposal seeks to study the reactivity of several late first-, second- and third-row transition metal ions with small hydrocarbons as a function of electronic excitation of the metal. Low energy ion drift cell and electronic state chromatographic methods will be used to study reactive systems such as platinum ion and methane. A successful outcome of this proposal would be a model of transition metal reactivity that would allow better catalysts to be designed. Important issues in this model include considerations of electron configuration and conservation of electron spin. New results from the larger metals proposed will be interpreted in the light of the PI's previous results on first row chemistry.

Transition metal ions have the ability to break sigma bonds, and this property is most useful in converting hydrocarbons to other compounds. Understanding the mechanism of this process will allow design of new and better catalysts for use in chemical synthesis and the petroleum industry.

The approach used in this project takes advantage of high-level interest in graphics, animation and storytelling. The Java-based Animation: Building viRtual Worlds for Object-oriented programming in Community colleges (JABRWROC) project uses simulation and visualization in a 3D, interactive, animation environment to introduce fundamental, object oriented programming concepts to novice programmers. The objectives are to 1) decrease attrition in introductory programming courses in community colleges, where open enrollment is conductive to high numbers of "at-risk" students, 2) attract students to computer and technology-related majors, 3) improve computer literacy courses to meet computer FITness guidelines defined by the NRC, by reintroducing programming /problem-solving modules, and 4) provide professional development for community college faculty in the use of innovative technology. Implementation occurs in three phases: developing and modifying instructional materials including lab exercises for traditional and returning learners; pilot testing in on-campus and distance learning classes, and dissemination. The potential impact includes over 6000 community college students on three campuses over the 3 year period. Urban students, specifically women and minorities, will be the primary audience for these new course materials. The assessment and evaluation will include persistence and attrition rates, student achievement, attitude surveys and interviews with instructors and students.

This ATE Project project addresses three issues in computing technology education in community colleges: (1) the need to reduce first-year attrition rates in computer science and information technology programs, (2) the need to adapt innovative approaches for teaching introductory programming to community college students, and (3) the need to provide effective community college faculty development materials for training in using innovative approaches. In our previous ATE project, JABRWOC (#0302542), they studied the effects of teaching fundamental programming concepts with visualization in pre-CS1 courses in community colleges. In this project, the innovative approach tested in JABRWOC will be extended to include a transition from Alice to Java in CS1 courses in community colleges, using the new, more powerful version of the software. In addition, this project proposes to develop a peer mentoring model and workshop materials designed to support community college faculty as they learn to teach with this innovative approach. Through our evaluation efforts, we will study the effectiveness of this approach for retaining students in pre-CS1 and CS1 courses and the effectiveness of the peer mentoring model and workshop materials for community college instructors.

In this project supported by the Chemical Structure, Dynamics and Mechanisms Program of the Division of Chemistry, Professor William Taylor of the University of Central Arkansas and his undergraduate research team will examine the activation of sigma bonds in halogenated molecules by metal ions. The reactions of interest will be carried out in the gas phase using a selected ion drift cell reactor which will generate transition metal ions in both ground and excited electronic states using a sputtering glow discharge source. These ions will then be mass selected via a quadrupole mass filter and introduced into a drift cell containing low pressures of helium and the neutral reactant molecule of interest. Subsequent analysis of reaction products will be carried out using a second quadrupole. This instrument will also be used to characterize the metal ion electronic state distributions via electronic state chromatography (ESC). ESC will also be used to determine the state-specificity of reaction products. Neutral reaction species will include various halogenated methanes (CH3X, CF3X, where X =Cl, Br and I), SF5Cl and SF5Br, and others. The experiments are designed to reveal the dependence of reaction outcome (e.g., halogen or halide abstraction, HX elimination) on various parameters, including the electronic state of the metal species as well as structural characteristics of the reactant neutral. Temperature dependent kinetic studies of selected reactions will be aimed at identifying likely transition states. These experimental efforts will be complemented by ensity functional calculations of reaction potential energy surfaces.

Transition metal ions have the ability to break sigma bonds, and this property is most useful in converting hydrocarbons to other, more useful compounds. Understanding the mechanism of this process will allow design of new and better catalysts for use in chemical synthesis and the petroleum industry. In addition, the project will provide research opportunities to undergraduates in an area which they might not otherwise be exposed to. Undergraduates participating in this work benefit from a valuable research experience which serves them well in a variety of subsequent career choices including graduate/professional programs and science-related jobs.

Under the supervision of Dr. Emily Jones (PI), William T. Taylor (co-PI) will analyze skeletal remains of ancient horses to shed light on the origins of horse herding and riding in eastern Eurasia. The use of the domestic horse for food and transport has been linked with major social, political, and economic changes to human societies, but its origins in prehistory are poorly understood. This project will address these questions through demographic study to identify ancient herding practices, morphological analyses to establish evidence for horseback riding and/or driving, and radiocarbon dating. This research will improve understanding of the links between domestic horse use, ecology, and human behavior. In addition, the project will fund the completion of Mr. Taylor's dissertation at the University of New Mexico, and will lead to the creation of an open-access 3D database of ancient horse remains to preserve these data for research and education. The work will be shared with the international community through talks and peer-reviewed manuscripts in both Mongolian and English.

Mr. Taylor's research will test predictions that horse-riding, herding societies developed in response to new ecological opportunities during the 2nd millennium BCE, centuries earlier than some existing models for East Asia suggest. By comparing ancient horses and modern specimens with known histories, the co-PI will characterize the impacts of bridling and riding on the equine skull, and using a 3D scanner, develop quantitative techniques for identifying horse transport in archaeological contexts. By estimating the age and sex of horse specimens buried at late Bronze Age stone monuments in Mongolia, Mr. Taylor will evaluate the possibility of ancient horse herding between 1300 and 700 BCE. With the aid of a portable 3D scanner, he will analyze these archaeological specimens for pathological changes, and use these data to assess the hypothesis that the horses were ridden or used to pull chariots. Finally, using new and published radiocarbon dates, Mr. Taylor will produce a chronological model to link changes in ancient Mongolian horse use with key environmental and cultural developments in Eurasia. These complementary studies will help to develop explanatory models for the origins and spread of horse herding and riding, which can be applied to modern and ancient societies around the globe.

In this project supported by the Chemical Structure, Dynamics and Mechanisms Program of the Division of Chemistry, Professor William Taylor of the University of Central Arkansas and his undergraduate research team are studying reactions between small molecules and ions which both contain atoms of halogen elements (the halogens include fluorine, chlorine, bromine and iodine). The ions contain transition metal elements (for example copper, nickel or gold). Many important industrial processes today involve catalysts that contain transition metal atoms. The studies in Professor Taylor's laboratory thus inform the broader area of catalysis, which is a key part of the production of an estimated 90% of all chemical products. Catalytic processes have a tangible environmental benefit by either direct remediation of pollutants or by reducing the environmental burden of industrial chemical production. Undergraduates participating in this work benefit from a valuable research experience that increases retention rates in the sciences, and prepares them for a variety of career choices.

The project explores the influence of electronic modifications to metal centers via the ligand environment on bond activation in neutral molecules. Studies addressed in this project examine the chemistry of MX+ (M=Ni, Cu, Au; X=F, Cl, Br, I) with a number of halogenated compounds known to be potent greenhouse gases and ozone depleters. In addition, reactions of CuX+ and AuX+ (X=Cl, F) with methane are examined in order to explore low-energy processes leading to the functionalization of this abundant, yet relatively inert, chemical resource. All reactions are studied experimentally in the gas phase using a selected ion drift cell reactor which generates transition metal ions in a sputtering glow discharge source. These metal ions are then be chemically modified with the addition of the desired halogen, mass selected, and introduced into a drift cell containing the neutral reactant molecule of interest. Analysis of reaction products is carried out using a second quadrupole. Experimental results are complemented computationally with density functional calculations and MP2 theory to gain additional insights into possible reaction mechanisms.

Contact PD/PI: Taylor, W. Robert Admin-Core-001 (377) Project Summary / Abstract - Administrative Core The Administrative Core will provide structure to the Georgia Clinical & Translational Science Alliance (GaCTSA), evaluate GaCTSA's efforts, and use that data to continually improve performance. To meet these goals, the Administrative Core will: 1) Provide leadership, structure and organization, governance, and communication support to enable GaCTSA to fully capitalize on unique and complementary strengths of the academic, healthcare, and development partners of the GaCTSA, 2) Coordinate the reporting of data across Cores, Functions, and collaborating institutions; enhance existing informatics systems and analytics to improve metric collection; build innovative data systems and survey methodologies; disseminate evaluation reports to GaCTSA leadership and oversight committees; and adopt and develop innovative evaluation metrics and techniques (including the new Common Metrics) to produce a comprehensive, data-rich assessment of GaCTSA impact throughout Georgia, and 3) Commit to improving quality of clinical and translational research by enhancing existing institutional infrastructures to ensure participant safety and ethical human subject research, then applying a systematic approach to maximize the efficiency of operations and distribute that knowledge across the partner institutions. Project Summary/Abstract Page 629 Contact PD/PI: Taylor, W. Robert Admin-Core-001 (377) OMB Number: 4040-0001 Expiration Date: 06/30/2016 RESEARCH & RELATED Senior/Key Person Profile (Expanded) PROFILE - Project Director/Principal Investigator Prefix: First Name*: W. Middle Name Robert Last Name*: Taylor Suffix: M.D. Position/Title*: Professor of Medicine Organization Name*: Emory University Department: Division: Street1*: Cardiology Division Street2: Emory University City*: Atlanta County: State*: GA: Georgia Province: Country*: USA: UNITED STATES Zip / Postal Code*: 303220000 Phone Fax Number: 404-727-3585 E-Mail*: wtaylor@emory.edu Number*: 404-727-8921 Credential, e.g., agency login: BKKMCTM Project Role*: Other (Specify) Other Project Role Category: Core Lead Degree Type: MD,PHD,BA Degree Year: File Name Mime Type Attach Biographical Sketch*: Attach Current & Pending Support: Page 630 Tracking Number: GRANT12172810 Funding Opportunity Number: PAR-15-304 . Received Date: 2016-05-25T16:47:02.000-04:00

PROJECT ABSTRACT The Georgia Clinical and Translational Science Alliance (Georgia CTSA) is a compelling partnership of Emory University, Morehouse School of Medicine (MSM), the Georgia Institute of Technology and the University of Georgia (UGA). The KL2 Program is an essential component of the Georgia CTSA and will respond to these critical needs to train a new and diverse generation of clinical and translational research (CTR) investigators who will carry out high impact research and translate new discoveries to improve health. The KL2 Program is focused on enhancing the career development of a diverse and talented group of junior faculty (Instructor or Assistant Professor level) with either PharmD, PhD and/or MD degrees at the Georgia CTSA partner institutions. Dr. Dunn will use the Georgia CTSA KL2 Program to obtain personalized training by completing carefully planned didactic coursework, individualized instruction, attending seminars and conferences, and completing supervised research experiences under the guidance of a mentoring team of seasoned investigators with expertise in depression physiology, neuroendocrine and stress physiologic pathways, biostatistical microbiome analysis, and translational research methods. The objective of this proposal is to explore biological pathways underpinning Postpartum depression (PPD)-the most common peripartum mood disorder affecting more than 500,000 women in the US annually. The latest evidence suggests that several biological mechanisms influence PPD symptoms including altered immune and inflammatory pathways. One area of scientific inquiry that has been found to influence immune and inflammatory pathways that has also emerged as a key depression biomarker and target for therapeutic intervention in non-pregnant human and animal models is the brain-gut microbiota axis (BGMA), an unexplored area of research within the context of PPD. Recent findings have shown that individuals with major depression demonstrate a different gut microbial composition than non-depressed individuals. Gut associated microbes are known to leak across the gut initiating inflammatory immune reactions in the gut mucosa as well as the host systemic inflammatory response. However, neither gut microbiome composition or gut leakiness ? or their contribution to host inflammation --have been explored in the context of PPD symptoms. Thus, to investigate the contribution of the BGMA on postpartum depressive symptoms in accordance with clinical postpartum care guidelines (within the first 3 weeks postpartum), DSM-V criteria (prenatal onset/ within 4 weeks following delivery), and recent Georgia maternal mortality data findings, we will prospectively enroll a pilot sample of 60 women at 3 weeks postpartum. We will investigate the relationships between the gut microbiome composition, markers of intestinal barrier dysfunction (LPS binding protein), systemic inflammatory markers (cytokines), and postpartum depressive symptoms. We will also measure and control for factors that have the potential to disturb the BGMA (diet, sleep, stress, and medications).