Analysis of Distal-less Gene Expression in Ciona intestinalis

Several homeobox-containing genes related to Distal-less (Dlx/Dll) have been isolated from a wide variety of organisms and have been shown to function as developmental regulators. In Drosophila only one Distal-less gene has been identified so far, and in vertebrates many components of the Distal-less family have been characterized. This suggests that, during the evolution of the Chordate phylum, the Dlx genes arose from an ancestral Distal-less gene via gene duplication. Three Dll homeoboxes have been isolated from the protochordate Ciona intestinalis and their clustered arrangement has been described. Since Ciona is regarded as one of the most primitive extant chordates, the present analysis gives us an insight into how these fundamental biological genes are evolved or are conserved during chordate evolution. The goal of this project was to clone coding sequences from the DllA/DllB cluster of Ciona intestinalis in order to observe the gene expression using in situ hybridization. During my study of DllA and DllB genes in Ciona, I used various techniques including PCR, restriction digest, sticky-end ligation, and transformation, to obtain cDNA from each gene. These cDNAs, DNA copies of the mRNA sequences, were used to create digoxigenin-labeled antisense RNA probes by in vitro transcription with T7 RNA polymerase. The in-situ hybridization documented gene expression in the epithelial cells and neural tissue, which is consistent with the expression patterns found in other species. The DllA probe was expressed in the anterior epithelial tissue in mid-tail stage embryos, on the adhesive papillae of late-tail stage embryos, and the bilateral primordia of the atrial siphon of larvae. DllB probe in blastula through early tail stages showed expression patterns in epidermal and neuroectodermal cells

Enhancing Attendance of the Greater Burlington YMCA Diabetes Fitness Program

Introduction: Since 1999, the Burlington YMCA has offered a free aerobics class to members of the community with diabetes. Additionally, the class regularly monitors participants’ health status. These data support the well-established benefit of exercise in diabetes. Medical Monitoring •hemoglobin A1C •blood pressure •resting pulse rate (before and after each twelve week session) About the Program •Began in 1999 •Free •Physician’s referral required •2h aerobics, twice per week: 1h land + 1h waterhttps://scholarworks.uvm.edu/comphp_gallery/1001/thumbnail.jp

MyChart Messaging: Patient Preferences about Timing of Provider Responses to Medical Advice Requests

Background: Patients frequently utilize portal messaging to reach their healthcare provider.1 This mode of communication allows for greater patient engagement in health concerns.2 MyChart messaging has grown recently, increasing physician workload.3,4 Understanding patient preferences for MyChart messaging may improve patient communication, potentially decreasing the response burden on physicians. Methods: A 12-question REDCap survey was sent via MyChart message to 31,502 patients in 8 UVM Health Network (UVMHN) affiliated adult primary care and family medicine practices. Survey data were analyzed in excel. Key Results: The survey was completed by 2011 (6.4%) patients, with 77% of respondents having used MyChart to ask their provider a question. MyChart messaging is always or often (69.1%) the first method patients use to contact their healthcare provider with 22% submitting urgent concerns. There is a significant association between when a patient prefers to receive a message and the time it takes providers to respond to their message (p \u3c 0.001). On average, patients would prefer to not restrict MyChart messaging to business hours. Discussion: Patients prefer prompt message responses from their providers and providers are meeting this preference. Future work could better observe the relationship between provider response time and patient preferences for physician response time. Data from Epic should be analyzed to determine if patient perceptions align with MyChart message records. Other efforts should include educating patients about appropriate messages to send via MyChart, or if a phone call is warranted. 1 North F, Luhman KE, Mallmann EA, Mallmann TJ, Tulledge-Scheitel SM, North EJ, Pecina JL. A Retrospective Analysis of Provider-to-Patient Secure Messages: How Much Are They Increasing, Who Is Doing the Work, and Is the Work Happening After Hours? JMIR Med Inform. 2020 Jul 8;8(7):e16521. doi: 10.2196/16521. 2 Avdagovska M, Ballermann M, Olson K, Graham T, Menon D, Stafinski T. Patient Portal Implementation and Uptake: Qualitative Comparative Case Study. J Med Internet Res. 2020;22(7):e18973. 3 Arndt BG, Beasley JW, Watkinson MD, et al. Tethered to the EHR: Primary Care Physician Workload Assessment Using EHR Event Log Data and Time-Motion Observations. Ann Fam Med. 2017;15(5):419-426. 4 Laccetti AL, Chen B, Cai J, et al. Increase in Cancer Center Staff Effort Related to Electronic Patient Portal Use. J Oncol Pract. 2016 (12):e981- e990

Provider Perspectives on Language Barriers in MyChart Enrollment for Patients with Limited English Proficiency

Background: There are 25 million people in the United States with limited English proficiency (LEP). Patients with LEP are a vulnerable population who may have worse health outcomes compared to English-proficient patients, as they face barriers to safe, effective, and high-quality care. Previous studies show this population is likely to benefit from utilization of patient portals, which allow users to access personal health information and communicate with healthcare providers. Current literature reports that patients with LEP and their families have a strong interest in using the patient portal as it can help them understand their providers and health, remember their care plan, and have more control over their health. However, there is less portal access and utilization among patients with LEP. This study was conducted to obtain provider perspectives surrounding MyChart engagement of patients with LEP to identify potential barriers to MyChart enrollment at the University of Vermont Health Network (UVMHN). Methods: EPIC’s Slicer Dicer tool was utilized to obtain UVMHN patient counts organized by reported language and MyChart activation. Data was collected from an anonymous REDCap survey sent to 211 providers within UVMHN Departments of General Internal Medicine, Family Medicine, and Pediatrics, and analyzed utilizing Excel. Results: 67.9% of UVMHN Primary Care English speakers have MyChart activated compared to 46.9% of patients with LEP. Providers strongly believe that MyChart is beneficial to patients and families with 72.1% of providers agreeing that it improves the patient-provider relationship. When asked specifically about patients with LEP, 36% of providers agree that MyChart access is beneficial to care. 49% report they are likely to encourage MyChart enrollment to their patients with LEP; however, only 14% know how to assist in enrollment. Differences in knowledge of language services are seen in different visit modalities: 78.1% of providers report ability to provide language services during in-person visits compared to 48.8% for video visits and 46.3% for telephone visits. Once made aware of the differences in MyChart activation status between English proficient patients and patients with LEP, providers frequently requested more education around language assistance, MyChart activation instructions to provide patients in their dominant language, translation tools within EPIC, and training for support staff to also encourage patients with LEP to enroll in MyChart. Conclusions: While providers overwhelmingly believe MyChart is useful to patient care and are likely to recommend it to their patients with LEP, they lack the tools and education necessary to aid in equitable enrollment. Educational materials and translated resources should be created for providers to utilize in assisting their patients with LEP. A follow-up study with patients would provide an opportunity to compare patient and provider responses and gain additional perspectives on MyChart access and utilization

Non-overlapping expression patterns of the clustered Dll-A/B genes in the ascidian Ciona intestinalis

The Ci-Dll-A and Ci-Dll-B genes of Ciona intestinalis are arranged in a convergently transcribed gene cluster. This genomic arrangement is similar to that of the multiple bigene clusters of the Dlx homologs in vertebrates. Analysis of whole genome sequences showed that linkage to the Hox cluster is conserved with the vertebrate clusters. Phylogenetic analysis supports gene trees consistent with homology of the ascidian and vertebrate Dlx clusters, and in combination with the apparent conservation of genomic arrangement, it is concluded that the ascidian cluster is most likely homologous with the vertebrate clusters. Using whole-mount in situ hybridization, Ci-Dll-B transcripts were detected in all ectodermal lineages through gastrulation. Expression is radically downregulated in the neurula with detectable expression disappearing around the time that Ci-Dll-A expression appears in the anterior ectoderm. By the late tailbud stage Ci-Dll-A transcripts were detected in the bilateral atrial primordia and persisted in the atrial rudiments to the larval stage, suggesting a role in development of these neural placode-like structures. This non-overlapping expression contradicts a common pattern seen in clustered genes, where as adjacent paralogs have largely overlapping expression domains. Enhancer sharing is often proposed as an explanation for the overlapping expression of gene cluster members. For this case of non-overlapping expression a model is proposed in which repressors acting at different stages override one or more shared enhancers. The enhancer sharing prevents breakup of the cluster while the independent temporal suppressors hide the presence of the shared enhancers. © 2007 Wiley-Liss, Inc

Nitric Oxide Isoenzymes Regulate Lipopolysaccharide-Enhanced Insulin Transport across the Blood-Brain Barrier

Insulin transported across the blood-brain barrier (BBB) has many effects within the central nervous system. Insulin transport is not static but altered by obesity and inflammation. Lipopolysaccharide (LPS), derived from the cell walls of Gram-negative bacteria, enhances insulin transport across the BBB but also releases nitric oxide (NO), which opposes LPS-enhanced insulin transport. Here we determined the role of NO synthase (NOS) in mediating the effects of LPS on insulin BBB transport. The activity of all three NOS isoenzymes was stimulated in vivo by LPS. Endothelial NOS and inducible NOS together mediated the LPS-enhanced transport of insulin, whereas neuronal NOS (nNOS) opposed LPS-enhanced insulin transport. This dual pattern of NOS action was found in most brain regions with the exception of the striatum, which did not respond to LPS, and the parietal cortex, hippocampus, and pons medulla, which did not respond to nNOS inhibition. In vitro studies of a brain endothelial cell (BEC) monolayer BBB model showed that LPS did not directly affect insulin transport, whereas NO inhibited insulin transport. This suggests that the stimulatory effect of LPS and NOS on insulin transport is mediated through cells of the neurovascular unit other than BECs. Protein and mRNA levels of the isoenzymes indicated that the effects of LPS are mainly posttranslational. In conclusion, LPS affects insulin transport across the BBB by modulating NOS isoenzyme activity. NO released by endothelial NOS and inducible NOS acts indirectly to stimulate insulin transport, whereas NO released by nNOS acts directly on BECs to inhibit insulin transport