Accelerated and efficient neuronal differentiation of Sox1GFP mouse embryonic stem cells in vitro using nicotinamide

A major challenge for advancement of clinical neuronal replacement therapies is the production of high yields of purified neuronal populations of appropriate phenotype with control over proliferation to prevent tumorigenesis. We previously reported that treatment of mouse embryonic stem cell (mESC;46CSox1GFP reporter cell line) monolayer cultures with the vitamin B3 metabolite nicotinamide at the early onset of development not only increased the efficiency of neuronal generation by two-fold but also enriched the ratio of purified neurons to non-neuronal cells in culture. This study aimed to investigate if nicotinamide enhances neural induction in this model and whether it also promotes the production/differentiation of specific neuronal subtypes. To address these aims, monolayer mESC cultures were treated with nicotinamide (10 mM) for different durations and immunocytochemistry/fluorescence microscopy was performed to assess the expression of stem cell, neural progenitor (NP) and neuronal subtype markers. Morphometric analyses were also performed to assess the extent of differentiation. Nicotinamide treatment significantly decreased Oct4+ pluripotent cells and concomitantly increased GFP+ cells at day 4, suggesting enhanced neural lineage commitment. By day 14, nicotinamide treatment (from day 0-7) reduced both Oct4+ and GFP expression concomitant with enhanced expression of neuron-specific β-tubulin, indicative of accelerated neuronal differentiation. Nicotinamide selectively enhanced the production of catecholaminergic, serotonergic and GABAergic neurons and, moreover, enhanced various aspects of neuronal morphology and maturation. Collectively, these data demonstrate a direct effect of nicotinamide at the initial stages of embryonic stem cell differentiation which could be critical for rapidly andefficiently promoting neural commitment to highly enriched neuronal lineages. The strong clinical potential of nicotinamide could successfully be applied to future neural cell-based therapies including Parkinson’s and Huntington’s disease, both to eradicate proliferating cells and for a more enhanced and specific differentiatio

A closer look into empathy among medical students: The career eulogy as a lens

Background Empathy is widely considered to be key to being an effective physician. The measurement of empathy is important to those designing medical education. The majority of the literature on empathy is based on survey scales that ask the learner to express their degree of agreement with a series of statements. We have previously studied and published an entirely projective measure that we term a career eulogy. Methods We had 65 students based at a regional rural medical school campus complete measures of a career eulogy (CE) and the Jefferson Scale of Empathy (JSE) over their four years in medical school. We then calculated weighted correlations between these two instruments. We also asked students to rank 10 factors that they thought affected student responses about empathy. Results We found a significant moderate correlation of JSE score with mentions of compassion on the CE (r = 0.414, p= 0.001). We also found that women scored higher on both instruments. The only factor showing consensus among students was that the general outlook on life was the most likely factor explaining student empathy responses. Conclusions Mentions in the compassion category on the CE appear to be measuring a concept very similar to empathy on the JSE. Students expressed that factors affecting their responses about empathy are very individual and that only interventions to change the general outlook on life may affect these measures of empathy. Having used the CE for the last five years, we find it to be a brief, very useful exercise both for measurement of empathy and as a group facilitation method in our professional identity curriculum. We welcome others to use our CE instrument in larger and more diverse groups to determine its true value in both measuring empathy and facilitating group process

New approaches to investigating the function of mycelial networks

Fungi play a key role in ecosystem nutrient cycles by scavenging, concentrating, translocating and redistributing nitrogen. To quantify and predict fungal nitrogen redistribution, and assess the importance of the integrity of fungal networks in soil for ecosystem function, we need better understanding of the structures and processes involved. Until recently nitrogen translocation has been experimentally intractable owing to the lack of a suitable radioisotope tracer for nitrogen, and the impossibility of observing nitrogen translocation in real time under realistic conditions. We have developed an imaging method for recording the magnitude and direction of amino acid flow through the whole mycelial network as it captures, assimilates and channels its carbon and nitrogen resources, while growing in realistically heterogeneous soil microcosms. Computer analysis and modeling, based on these digitized video records, can reveal patterns in transport that suggest experimentally testable hypotheses. Experimental approaches that we are developing include genomics and stable isotope NMR to investigate where in the system nitrogen compounds are being acquired and stored, and where they are mobilized for transport or broken down. The results are elucidating the interplay between environment, metabolism, and the development and function of transport networks as mycelium forages in soil. The highly adapted and selected foraging networks of fungi may illuminate fundamental principles applicable to other supply networks

Keeping Rural Medical Students Connected to their Roots: A “Home for the Holidays” Immersion Experience

Keeping Rural Medical Students Connected to their Roots: A “Home for the Holidays” Immersion Experience Abstract Introduction Preclinical medical students with an initial affinity for rural practice often experience urban disruption during training. Longer summer rural immersion experiences have been shown to be somewhat effective in keeping rural students connected to their rural roots. Some students cannot adapt their schedules to these longer experiences. In response, a brief practice experience near home during the winter break of the academic year was established. Methods Nineteen detailed rural practice assessments were completed by preclinical medical students during school years 2009-2014 and essays and anonymous evaluations of the experience were analyzed. Results All students reported the four components of the experience increased understanding of rural medicine and all but 2 reported it was relevant to their career objectives. These two, at this early stage in training, decided rural practice was not for them and subsequently chose sub-specialty training. Six of the 12 (50%) who have completed medical school chose a family medicine residency. The themes in the essay portion included insights into rural lifestyle, returning home, health system organization issues, and patient expectations and traditions. Conclusions Initial experience with a “home for the holidays” approach for rural immersion of rural medical students was successful in reconnecting these students to their roots, and 17/19 (90%) reported their intention to choose rural practice

A Career Eulogy Reflective Exercise: A View into Early Professional Identity Formation

Beginning medical students have a very early idea of what their physician identity will be. Providing a brief structured opportunity to reflect on the end of their career can be an important first step in identity formation. A reflective exercise was used in the summer prior to beginning medical school as each class of students at a regional rural medical school began a summer prematriculation program from 2015-2019. Students wrote what they wished to be said about them at the end of their career using a “Career Eulogy.” Identifiers were removed and narratives were coded into recurring text clusters by the authors. The students, on an anonymous evaluation, strongly agreed that the exercise facilitated the accomplishment of the program goals. Reflections from 42 entering medical students indicated a preference to be remembered for compassion, passion, quality, and patient relationships. Men more frequently mentioned family and enjoying life, and women more frequently mentioned patient relationships. Rural students included quality and being a teacher of medical students more frequently, and those with a physician parent included passion and community more frequently. Those who later chose family medicine as a specialty more often included references to enjoying life and family. This reflective exercise provided useful insight into early professional identity formation among these medical students and served to encourage group discussion. It could be used with larger, more diverse groups to determine its value and clarify true differences among the demographic and specialty choice patterns. This exercise could also be used annually for each student, providing a longitudinal view of identity formation

Do medical residents with rural upbringing show less decline in empathy during training? A report from a rural family medicine residency

Abstract Purpose Our focus in this study was to determine if demographic variables, including specifically rural upbringing, showed any association with a measure of empathy among family medicine residents at a rural site. Methods We surveyed 40 residents annually using the Jefferson Scale of Empathy (JSE) from 2016 to 2020 and compared scores between residents with rural vs urban hometowns. The response rate was 98/99 (99%) of completed survey opportunities. Findings There was no significant difference in JSE scores of rural vs urban residents at baseline, and both groups showed a decline after the PG-1 year. However, the rural-raised residents showed a significantly different rebound after the PG-2 and PG-3 year, with the urban-raised residents actually showing a slight continuing decline (p=.023 and p=.033). Conclusion These preliminary findings among family medicine residents at a rural site suggest that rural background residents might regain empathy better than urban background residents during the course of their training. Further study should validate our findings and address possible explanations, including the importance of cultural concordance with the patient population served. This and other hypotheses will be explored in further studies with focus groups and other contemporaneous measures

Rural Medical Student Opinions About Rural Practice: Does Choice of College Make a Difference?

Purpose While many programs assume medical students who attended rural colleges are more likely to practice in smaller towns later, there are few studies to support this assumption. This study examines aspects of medical students’ opinions about rural living and rural practice based on where they attended college. Method Students with rural upbringing were identified and administered a previously published rural health opinion survey. The opinion survey was administered across three academic years, from 2015 to 2017, and included a total of three scales and nine subscales for analysis. Factor analysis was used to identify the nine subscales. Results Respondents who attended small town colleges tended to have much stronger opinions about the positive aspects of small town living. Small town college attendees also had stronger positive opinions regarding rural practice than their larger college counterparts. Discussion These preliminary data indicate considering a medical school applicants’ choice of college may provide insight into their opinions about future rural practice. Additional research is needed to collect a larger dataset to examine the association of college choice with opinions about rural practice

Zebrafish Cytosolic Carboxypeptidases 1 and 5 Are Essential for Embryonic Development

The cytosolic carboxypeptidases (CCPs) are a subfamily of metalloenzymes within the larger M14 family of carboxypeptidases that have been implicated in the post-translational modification of tubulin. It has been suggested that at least four of the six mammalian CCPs function as tubulin deglutamylases. However, it is not yet clear whether these enzymes play redundant or unique roles within the cell. To address this question, genes encoding CCPs were identified in the zebrafish genome. Analysis by quantitative polymerase chain reaction indicated that CCP1, CCP2, CCP5, and CCP6 mRNAs were detectable between 2 h and 8 days postfertilization with highest levels 5–8 days postfertilization. CCP1, CCP2, and CCP5 mRNAs were predominantly expressed in tissues such as the brain, olfactory placodes, and pronephric ducts. Morpholino oligonucleotide-mediated knockdown of CCP1 and CCP5 mRNA resulted in a common phenotype including ventral body curvature and hydrocephalus. Confocal microscopy of morphant zebrafish revealed olfactory placodes with defective morphology as well as pronephric ducts with increased polyglutamylation. These data suggest that CCP1 and CCP5 play important roles in developmental processes, particularly the development and functioning of cilia. The robust and similar defects upon knockdown suggest that each CCP may have a function in microtubule modification and ciliary function and that other CCPs are not able to compensate for the loss of one

Strategic Scheduling of Infrastructure Repair and Maintenance: Volume 1—Decision Tree for Steel Bridge Painting

Even for the right treatment, improper timing can have consequences: premature application (treatment is applied too early) could mean wasteful spending even if users enjoy the benefits of higher pavement condition; deferred or delayed application (treatment is applied too late) could result in higher user costs due to poor condition and even reduced asset longevity. The objectives of this research were to establish the optimal condition or timing for each of the standard M&R treatment types typically used by INDOT; quantify the consequences of departures from such optimal conditions or timings; and to establish the optimal M&R treatment schedule for each asset family. The study focused on three asset types and their treatments: Painting of Steel Bridges. A painting decision tree was developed, to serve as a framework that would enable INDOT to consider other maintenance treatment types, namely spot repair/painting and overcoating. Bridge Deck Maintenance and Rehabilitation. Life-cycle condition-based deck M&R strategies based on different trigger results were proposed and presented. Pavement Maintenance, Rehabilitation, and Replacement. A framework was established to find the optimal scheduling for multiple treatments and recommend appropriate long-term M&R strategies for flexible and rigid pavements on different functional classes