Although remnant cardiomyocytes (CMs) possess a certain degree of proliferative ability, efficiency is too low for cardiac regeneration after injury. In this study, we identified a distinct stage ...within the initiation phase of CM reprogramming before the MET process, and microarray analysis revealed the strong up‐regulation of several mitosis‐related genes at this stage of reprogramming. Several candidate genes were selected and tested for their ability to induce CM proliferation. Delivering a cocktail of three genes, FoxM1, Id1, and Jnk3‐shRNA (FIJs), induced CMs to re‐enter the cell cycle and complete mitosis and cytokinesis in vitro. More importantly, this gene cocktail increased CM proliferation in vivo and significantly improved cardiac function and reduced fibrosis after myocardial infarction. Collectively, our findings present a cocktail FIJs that may be useful in cardiac regeneration and also provide a practical strategy for probing reprogramming assays for regeneration of other tissues.
Synopsis
The proliferation rate of remnant cardiomyocytes (CMs) is too low for cardiac regeneration after injury. Here, a gene cocktail is defined to induce CMs to efficiently proliferate, while improving cardiac function after myocardial infarction in mice.
The second day prior to the mesenchymal–epithelial transition was identified as the critical time for initiating cardiomyocyte reprogramming.
Microarray data analysis pinpointed a set of up‐regulated genes that were used to derive a reprogramming gene cocktail.
The gene cocktail recipe comprising FoxM1, Id1, and Jnk3‐shRNAs (FIJs) enhances cardiomyocyte proliferation.
Cardiomyocyte cell cycle re‐entry is followed by completed mitosis and cytokinesis to execute a delayed reprogramming.
The FIJs gene cocktail improves heart function after myocardial infarction in a mouse model.
The proliferation rate of remnant cardiomyocytes (CMs) is too low for cardiac regeneration after injury. Here, a gene cocktail is defined to induce CMs to efficiently proliferate, while improving cardiac function after myocardial infarction in mice.
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FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
Data-independent acquisition modes isolate and concurrently fragment populations of different precursors by cycling through segments of a predefined precursor m/z range. Although these selection ...windows collectively cover the entire m/z range, overall, only a few per cent of all incoming ions are isolated for mass analysis. Here, we make use of the correlation of molecular weight and ion mobility in a trapped ion mobility device (timsTOF Pro) to devise a scan mode that samples up to 100% of the peptide precursor ion current in m/z and mobility windows. We extend an established targeted data extraction workflow by inclusion of the ion mobility dimension for both signal extraction and scoring and thereby increase the specificity for precursor identification. Data acquired from whole proteome digests and mixed organism samples demonstrate deep proteome coverage and a high degree of reproducibility as well as quantitative accuracy, even from 10 ng sample amounts.
Biofluids contain molecules in circulation and from nearby organs that can be indicative of disease states. Characterizing the proteome of biofluids with DIA-MS is an emerging area of interest for ...biomarker discovery; yet, there is limited consensus on DIA-MS data analysis approaches for analyzing large numbers of biofluids. To evaluate various DIA-MS workflows, we collected urine from a clinically heterogeneous cohort of prostate cancer patients and acquired data in DDA and DIA scan modes. We then searched the DIA data against urine spectral libraries generated using common library generation approaches or a library-free method. We show that DIA-MS doubles the sample throughput compared to standard DDA-MS with minimal losses to peptide detection. We further demonstrate that using a sample-specific spectral library generated from individual urines maximizes peptide detection compared to a library-free approach, a pan-human library, or libraries generated from pooled, fractionated urines. Adding urine subproteomes, such as the urinary extracellular vesicular proteome, to the urine spectral library further improves the detection of prostate proteins in unfractionated urine. Altogether, we present an optimized DIA-MS workflow and provide several high-quality, comprehensive prostate cancer urine spectral libraries that can streamline future biomarker discovery studies of prostate cancer using DIA-MS.
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IJS, KILJ, NUK, PNG, UL, UM
The purpose of this grant proposal was to develop and seek funding for implementation of an art therapy program for adults with developmental disabilities in Orange County, California. The clients of ...Westview Services' adult day programs located in Orange County will have an opportunity to participate in the proposed art therapy program. Through a review of the literature on the needs and challenges as well as current services and interventions in place for individuals with developmental disabilities, the grant writer proposed and designed an art therapy program. This grant writer explored potential public and private funding sources, which yielded the Field Initiated Projects through the National Institute on Disability and Rehabilitation Research as the funding source with the most potential fitting the goals and objectives of this project. The actual submission and/or funding of this grant were not required for the successful completion of this academic project. Copies of dissertations may be obtained by addressing your request to ProQuest, 789 E. Eisenhower Parkway, P.O. Box 1346, Ann Arbor, MI 48106-1346. Telephone 1-800-521-3042; e-mail: disspub@umi.com
Riboswitches are metabolite-sensitive elements found in mRNAs that control gene expression through a regulatory secondary structural switch. Along with regulation of lysine biosynthetic genes, ...mutations within the lysine-responsive riboswitch (L-box) play a role in the acquisition of resistance to antimicrobial lysine analogs. To understand the structural basis for lysine binding, we have determined the 2.8Å resolution crystal structure of lysine bound to the Thermotoga maritima asd lysine riboswitch ligand-binding domain. The structure reveals a complex architecture scaffolding a binding pocket completely enveloping lysine. Mutations conferring antimicrobial resistance cluster around this site as well as highly conserved long range interactions, indicating that they disrupt lysine binding or proper folding of the RNA. Comparison of the free and bound forms by x-ray crystallography, small angle x-ray scattering, and chemical probing reveals almost identical structures, indicating that lysine induces only limited and local conformational changes upon binding.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
The purpose of this grant proposal was to develop and seek funding for implementation of an art therapy program for adults with developmental disabilities in Orange County, California. The clients of ...Westview Services' adult day programs located in Orange County will have an opportunity to participate in the proposed art therapy program. Through a review of the literature on the needs and challenges as well as current services and interventions in place for individuals with developmental disabilities, the grant writer proposed and designed an art therapy program. This grant writer explored potential public and private funding sources, which yielded the Field Initiated Projects through the National Institute on Disability and Rehabilitation Research as the funding source with the most potential fitting the goals and objectives of this project. The actual submission and/or funding of this grant were not required for the successful completion of this academic project.
The primary purpose of this study was to determine the degree to which Nipomo Healthy Start, a school-based mental health program, was able to meet the goals and objectives outlined in its original ...grant proposal to the state of California, and to determine what barriers prevent service delivery to students and families. Barriers to service delivery were determined using a survey which was given to the staff. Staff answered questions on a 7-point Likert scale about the frequency a specific item had been a barrier to service within the last year. Results indicated that barriers rooted in the family (M=3.93) were more frequently encountered than barriers rooted within the agency (M=3.55). The goals of the program were evaluated using primarily archival data in the form of annual reports made to the state, and in interviews with stakeholders, staff, and teachers. No statistically significant results were found for the program’s target outcomes, but there were improvements in most areas for the three school sites. This evaluation determined that Nipomo Healthy Start may not have met each target outcome as measured by its indicator in the grant proposal, but that the program’s responsiveness to the community’s needs, its continued flexibility, and commitment to the youth of Nipomo has created a critical and much used resource in the community.
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