Progress 10/01/12 to 07/01/13
Outputs
Target Audience: The research is being prepared for publication but was not presented at meetings during the last reporting period. The target audience was the same as for other reporting periods, both academic scientists and others interested in food safety. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Dr. Threadgill's postdoc Dr. Kristen Delaney has been able to move to an independent faculty position at Fayetteville State University as a result of her training and professional development as a result of working on this project. How have the results been disseminated to communities of interest? Once the manuscript is accepted for publication, the results will be available to communities of interest. The manuscript will be submitted before the holiday shutdown. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Experiments in C. jejuni have been completed to submit the first publication. Some additional studies with E. coli are proposed to extend the results. The initial results with E. coli unfortunately did not yield the expected results and thus experimental methods will need to be reviewed and revised. Both the primary laboratory (Dr. Threadgill, NCSU to Texas A&M) and the collaborating laboratory (Dr. Jobin, UNC to UF) have moved to new locations. These new laboratories will be restarted and the work continued from the new locations.
Publications
|
Progress 10/01/11 to 09/30/12
Outputs
OUTPUTS: Activities were as follows: Additional experiments were completed to finalize a publication for this project. We have obtained a second C. jejuni strain mutated for the Cj 0372 gene. We have obtained E. coli mutants for a similarly annotated gene and expressed both Cj 0372 and Cj 0371. Antibodies to both of these proteins have been generated. We have verified that Cj0371 is produced at normal levels in the Cj 0372 mutant and thus demonstrated that the loss of Cj 0372 is responsible for the phenotypes we have measured. Posters on the research were presented at the NC ASM meeting Oct. 2011 by both a postdoc and undergraduate student. The postdoc also presented a poster on this work at the 16th Intl. Campylobacter, Helicobacter and Related Organisms Mtg, 2011. Our own efforts have led to a soon to be completed independent publication. Through our collaborators at UNC a related publication was successfully completed. PARTICIPANTS: Participants: PI: Dr. Deborah Threadgill, Post-doctoral fellow: Dr. Kristen Delaney, Research Associate: Ms. Erin Harrell (MS) and NCSU undergraduate student Ms. Bridget Conley. Collaborators: Dr. Christian Jobin, and Dr. Xiaolun Sun, UNC Chapel Hill. Training opportunities for Dr. Kristen Delaney as well as Ms. Bridget Conley. TARGET AUDIENCES: Target Audiences: The NC ASM and CHRO are public meetings where any interested party may register to attend. The knowledge gained is therefore available to the public prior to publication of completed research. PROJECT MODIFICATIONS: We are now able to work with two mutant C. jejuni strains that we created, and also to work with E. coli mutants to determine the precise function of the annotated genes. We have established new connections with labs that work on the E. coli enzyme to obtain reagents and protocols so that we can complete our C. jejuni studies.
Impacts
Change in knowledge: We have come to a clearer understanding about a previously unknown gene in C. jejuni that we saw was elevated in expression at the chicken body temperature. This gene appears to be a conserved gene related to coping with oxidative stress that is also present in E. coli and we are using E. coli mutants as well as gene expression clones from E. coli to verify the C. jejuni studies. Change in action: Recent publications from groups working with E. coli have indicated a clear path forward for our research, and we are confident our revised and expanded publication will now be accepted at a ranked journal. We can now assay the function of the C. jejuni gene using clear protocols for E. coli, and can also show that the gene has similar functions in C. jejuni to what it has in E. coli.
Publications
- Sun X, Threadgill DS, Jobin C. 2012. Campylobacter jejuni induces colitis through activation of mammalian target of rapamycin signaling pathway. Gastroenterology. 142 (86-95)
|
Progress 10/01/09 to 09/30/10
Outputs
OUTPUTS: Activities were as follows: Experiments were initiated to finalize a publication for this project. In particular, a manuscript had been submitted several years ago without successful acceptance by a journal. With the release of some of the promised start-up monies, this manuscript was able to be revisited and further critical experimentation instituted. Services: During this time the original first author of the paper (now an Asst. Professor at a nearby university) has been helping the new postdoc and myself with the research with the aim of resubmission in the last year of the project. I will be submitting for NSF funding in the 2011 fiscal year that will include this collaboration. PARTICIPANTS: PI: Dr. Deborah Threadgill, Postdoctoral fellow: Dr. Jinzhi Wang (NCSU), Collaborator: Dr. Jason Andrus Meredith University, Raleigh NC Collaborator: Dr. Christian Jobin, UNC-Chapel Hill Training opportunities for the postdoctoral fellow Dr. Wang whose original background is in food science rather than bacterial pathogenesis. Also training provided for one post-BS NCSU Microbiology major, and a current NCSU Microbiology major. TARGET AUDIENCES: We will be presenting a poster on this work at the American Society for Microbiology in May. This meeting is open to members of the society as well as the general public. The knowledge obtained is therefore available to the public prior to publication of the manuscript. PROJECT MODIFICATIONS: Major changes in the approach to the project have taken place because of the release of sufficient funds to allow the completion of the project. In particular, we can now express the protein that is lacking in the mutant strain and explore its function. These studies were not possible previously because of the unknown nature of the protein and the anticipated expense involved therefore in characterizing the function. Staffing for the lab also was problem and has been partially solved with the addition of Dr. Wang as well as the two students who have been involved.
Impacts
Change in knowledge: We have come to a deeper understanding about one of the genes identified during our studies of protein glycosylation in C. jejuni. This gene when mutated causes a number of phenotypic effects but we were unclear about the exact mechanism of its action. Now we have a clearer picture of what the gene's function is and this is helping us with the manuscript completion. Change in action: A new experimental direction to explore the mechanism involved with the unknown gene's function has been instituted. We are confident that this will enhance our ability to convince reviewers of the importance of our study results, since some of the prior criticisms were directed at the unknown nature of the genes involved and what they could be doing.
Publications
- No publications reported this period
|
Progress 10/01/08 to 09/30/09
Outputs
OUTPUTS: Although this research project was filed in October 2008, the laboratory renovations were not completed until March 2009. The start-up for this project was frozen in May 2009, thus the project could not be initiated as planned. The laboratory was set up with previously purchased equipment, but no needed supplies or necessary equipment could be purchased. Collaborative projects at other institutions were continued, and these contributed to the publication listed below. PARTICIPANTS: Dr. Deborah Threadgill, PI and Jennifer Lowther, Research Specialist were the NCSU participants. Ms. Lowther contributed to lab set up and minimal data generation as well as supervision of an undergraduate student worker (hired with NCI funds for another project). Dr. Threadgill supervised Ms. Lowther's activities and recommended additional training as appropriate. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Because of the lack of funding for this project, alternative funding sources for research were sought. In particular, the lab obtained funding from NCI for an unrelated project. Funding for another unrelated project was sought from NIDCR/NIAID, and will be revisited this fiscal year. The UNC collaboration with Dr. Christian Jobin also sought funding for C. jejuni research, and this will be revisited in the next fiscal year. Because there was no supply or equipment budget for the project, the research goals were severely attenuated. In particular, the goals were reduced to improve quality control of the bacterial strains that would be used by Dr. Jobin's group as well as Dr. Threadgill's research program once funded.
Impacts
Through a collaborative project carried out at UNC-Chapel Hill, results were obtained for a mouse model of C. jejuni infection. These results will be helpful to the goals for the outlined project once the funds for the project are restored.
Publications
- Lippert E, Karrasch T, Sun X, Allard B, Herfarth HH, Threadgill D, Jobin C 2009 Gnotobiotic IL-10; NF-kappaB mice develop rapid and severe colitis following Campylobacter jejuni infection. PLoS One.4(10):e7413.
|
|