Summary• CONSTANS is an evolutionarily-conserved central component of the genetic pathway that controls the onset of flowering in response to daylength. However, the specific biochemical mechanism by which the CONSTANS protein regulates the expression of its target genes remains largely unknown.• By using a combination of cell-based expression analysis and in vitro DNA binding studies, we have demonstrated that CONSTANS possesses transcriptional activation potential and is capable of directly binding to DNA.• CONSTANS was found to bind DNA via a unique sequence element containing a consensus TGTG(N2-3)ATG motif. This element is present in tandem within the FLOWERING LOCUS T promoter and is sufficient for CO binding and activity. The conserved CCT (CONSTANS, CONSTANS-like and TOC1) domain of CONSTANS was shown to be required for its recruitment to the DNA motif and other CCTcontaining proteins were also found to have the ability to regulate gene expression via this element.• The CCAAT box, which has been previously hypothesized as a recruitment site for complexes containing the CONSTANS protein, potentiated CONSTANSmediated activation but was not essential for CONSTANS recruitment to a target promoter or for its activity as a transcriptional factor.

Chemical inhibition of the mitochondrial electron transport chain (mtETC) by antimycin A (AA) or the TCA cycle by monofluoroacetate (MFA) causes increased expression of nucleus-encoded alternative oxidase (AOX) genes in plants. In order to better understand the mechanisms of this mitochondrial retrograde regulation (MRR) of gene expression, constructs containing deleted and mutated versions of a promoter region of the Arabidopsis thaliana AOX1a gene (AtAOX1a) controlling expression of the coding region of the enhanced firefly luciferase gene were employed to identify regions of the AtAOX1a promoter important for induction in response to mtETC or TCA cycle inhibition. Transient transformation coupled with in vitro and in vivo assays as well as plants containing transgenes with truncated promoter regions were used to identify a 93 base pair portion of the promoter, termed the MRR region, that was necessary for induction. Further mutational analyses showed that most of the 93 bp MRR region is important for both AA and MFA induction. Sub-regions within the MRR region that are especially important for strong induction by both AA or MFA were identified. Specific mutations in a W-box and Dof motifs in the MRR region indicate that these specific motifs are not important for induction. Recent evidence suggests that MRR of AOX genes following inhibition of the mtETC is via a separate signaling pathway from MRR resulting from metabolic shifts, such as those that result from MFA treatment. Our data suggest that these signaling pathways share regulatory regions in the AtAOX1a promoter. Arabidopsis proteins interacted specifically with a probe containing the MRR region, as shown by electrophoretic mobility shift assays and Southwestern blotting. These interactions were eliminated under reducing conditions.

When compared with the general United States population, Hispanic Americans are at an increased risk of developing type 2 diabetes mellitus (DM2) and are far more likely to suffer devastating complications related to the disease. The purpose of this quality improvement project was to determine whether the use of a culturally tailored, mobile application prototype educational tool increased DM2 prevention knowledge among Hispanic patients at risk for DM2. The educational tool contained information about DM2 including risk factors, prevention, and health maintenance. The prototype was developed to function like a working mobile application and a pre/posttest was administered to participants at three local Hispanic community health fairs in Cincinnati, Ohio. Paired t test analysis of the 27 completed surveys showed a statistically significant improvement in posttest scores. The results showed that the average score was 4.1 out of a total of five possible points in the pre-test. The mean total score of the post-test was 4.7, with a total improvement of the mean score of 0.6 (0.0001). It was concluded that there was a statistically significant improvement in the knowledge of DM2 prevention after reviewing the material presented in the application prototype. In addition, participants expressed a strong interest in a working mobile application that offers culturally tailored DM2 prevention education.