Leveraging digital assets: lessons from a 14-year-old isotope tracer course for professional scientists.

The COVID-19 pandemic and subsequent policies (e.g., social distancing, travel restrictions) challenged both organizers for and attendees of programs typically held in-person. Many scientific training programs quickly adapted to virtual formats by incorporating digital assets developed for virtual learning and remote social engagement. At the outset, the value of continuing digital elements with future in-person events was unclear. To examine how virtual resources supported heterogeneous professional training programs, we reviewed survey data for a 14-year-old training program for scientific professionals titled Isotope Tracers in Metabolic Research: Principles and Practice of Kinetic Analysis. We found a positive relationship between survey satisfaction and the post-pandemic, in-person program that included digital assets held in 2022 when compared to pre-pandemic in-person programs. To better understand the post-pandemic program satisfaction, we assessed the 2021 virtual course format and survey data. We found that although there was a desire to return to in-person programs, the digital assets and approaches were valued. In examining the individual programmatic elements in the 2022 in-person course, there was better "value and understandability" of lectures over previous in-person years. These findings highlight how incorporating new digital engagement strategies for professional development benefit even the most established programs in supporting heterogeneous learners.

How to design a broad mindfulness program: One approach to introducing mindfulness to a STEM commumity within a university setting.

Mindfulness has become popular in recent decades as a tool for psychological well-being. However, mindfulness has yet to find a solid footing as a routine practice within the Science, Technology, Engineering, and Mathematics (STEM) field. Here, we discuss the design of an introductory mindfulness program that provides the STEM community with a broad overview of various mindfulness methods. We also discuss delivery implementation methods and other considerations when designing mindfulness programming. This article provides resources for those interested in incorporating mindfulness into the STEM field, especially within the university setting.

How to design an art-science program? Self-reported benefits for artists and scientists in the VI4 artist-in-residence program.

While many new programs bridge the arts and sciences, a data-based examination of art-science program design can lead to more efficient programming. The Vanderbilt Institute for Infection, Immunology, and Inflammation Artist-in-Residence program is a virtual program that brings together undergraduate student "artists" and faculty-level "scientists" to generate science-art content. We have recruited over 80 artists and 50 scientists to collaborate in creating visual science communication content. Using self-reported data from both groups, we performed qualitative and quantitative analyses to define sources for negative and positive experiences for artists and scientists. We also identify areas for improvement and key features for in producing a positive experience. We found that artists participants had more positive responses about "learning something new" from the program than scientists. We also found that for both artists and scientists the length of the program and the virtual nature were identified as key features that could be improved. However, the most surprising aspect of our analysis suggests that for both "way of thinking" and "science communication to the public or general audience," were seen as significant beneficial gains for scientists compared to artists. We conclude this analysis with suggestions to enhance the benefits and outcomes of an art-science program and ways to minimize the difficulties, such as communication and collaboration, faced by participants and program designers.

Innovations in Undergraduate Research Training Through Multisite Collaborative Programming: American Heart Association Summer Undergraduate Research Experience Syndicate.

Background To support diversity in biomedical science, the American Heart Association launched the Supporting Undergraduate Research Experiences for undergraduate students from underrepresented backgrounds to provide mentorship and high-level exposure at 5 leading medical institutions. Here we describe the initial formation of the partnership and the alteration made in response to the program to accommodate COVID-19 safety precautions. Methods and Results We outline how programming shifted from local, in-person programming in the summer of 2019 to a collaborative, mainly virtual curriculum in 2020 using students' self-reported before and after surveys from both 2019 (n=33) and 2020 (n=42). Students from both in-person (2019) and virtual programs (2020) self-reported significant gains in scientific proficiency. A qualitative-directed content analysis of student open-response questions was performed. Students reported extensive benefits from the 2020 virtual training, including Personal Gains, Research Skills, Thinking and Working Like a Scientist, and Attitudes and Behaviors. Notedly, we observed increases in the Attitudes and Behaviors category. We outline the pros and cons of in-person and virtual programming and make recommendations moving forward in a postpandemic world with hybrid work and learning systems. Conclusions Our effort informs the development of future undergraduate research training programs, significantly maximizing a hybrid training modality. The American Heart Association Supporting Undergraduate Research Experiences serves as a model for building multi-institutional partnerships and providing research experiences that overcome institutional barriers and support students' interests, commitment, and ability to persist in science, technology, engineering, and math fields.

Virtual Vanderbilt Summer Science Academy highlighted the opportunity to impact early STEMM students career knowledge through narrative.

Due to COVID-19 precautions, the Vanderbilt University summer biomedical undergraduate research program, the Vanderbilt Summer Science Academy (VSSA), rapidly transitioned from offering an in-person training program to a virtual seminar format. Our program typically supports undergraduate development through research and/or clinical experience, meeting with individuals pursuing postgraduate training, and providing career development advice. Evidence supports the idea that summer programs transform undergraduates by clarifying their interest in research and encouraging those who haven't previously considered graduate studies. We were interested in exploring whether a virtual, synchronous program would increase participants' scientific identity and clarify postgraduate career planning. Rather than create a virtual research exposure, our 5-week "Virtual VSSA" program aimed to simulate the casual connections that would naturally be made with post-undergraduate trainees during a traditional summer program. In seminars, presenters discussed 1) their academic journey, explaining their motivations, goals, and reasons for pursuing a career in science as well as 2) a professional story that illustrated their training. Seminars included Vanderbilt University and Medical School faculty, M.D., MD/Ph.D., as well as Ph.D. students from diverse scientific and personal backgrounds. In addition, weekly informational sessions provided an overview of the nature of each degree program along with admissions advice. Through pre-and post-program surveys, we found that students who registered for this experience already strongly identified with the STEMM community (Science, Technology, Engineering, Mathematics, and Medicine). However, participation in the Virtual VSSA increased their sense of belonging. We also uncovered a gap in participants' understanding of postgraduate pathways prior to participation and found that our program significantly increased their self-reported understanding of postgraduate programs. It also increased their understanding of why someone would pursue a Ph.D. or Ph.D./MD versus M.D. These changes did not uniformly impact participants' planned career paths. Overall, by providing personal, tangible stories of M.D., MD/Ph.D., and Ph.D. training, the Virtual VSSA program offered seminars that positively impacted students' sense of belonging with and connection to the STEMM disciplines.

Production Processes for Creating Educational Videos.

Asynchronous video-based educational resources allow for increased course material engagement. In today's climate, educators are encouraged to create videos for online instruction but are typically given limited production guidance. Few formal resources exist to guide educators for high-quality video production in a non-studio setting. This article is a how-to guide for producing videos using widely available primary resources through three steps: preproduction, production, and postproduction. During preproduction, educators consider style and project scope, including the "what, how, and why" of the content. For production, we have provided information on the set, light, sounds, and video equipment needed for optimizing video production in a non-studio setting. Finally, during postproduction, the educator considers how to combine and edit the video as well as organize content. Overall, this article is an approachable guide to help educators begin their low-budget video-production journeys.

mGlu2 and mGlu3 Negative Allosteric Modulators Divergently Enhance Thalamocortical Transmission and Exert Rapid Antidepressant-like Effects.

Non-selective antagonists of metabotropic glutamate receptor subtypes 2 (mGlu2) and 3 (mGlu3) exert rapid antidepressant-like effects by enhancing prefrontal cortex (PFC) glutamate transmission; however, the receptor subtype contributions and underlying mechanisms remain unclear. Here, we leveraged newly developed negative allosteric modulators (NAMs), transgenic mice, and viral-assisted optogenetics to test the hypothesis that selective inhibition of mGlu2 or mGlu3 potentiates PFC excitatory transmission and confers antidepressant efficacy in preclinical models. We found that systemic treatment with an mGlu2 or mGlu3 NAM rapidly activated biophysically unique PFC pyramidal cell ensembles. Mechanistic studies revealed that mGlu2 and mGlu3 NAMs enhance thalamocortical transmission and inhibit long-term depression by mechanistically distinct presynaptic and postsynaptic actions. Consistent with these actions, systemic treatment with either NAM decreased passive coping and reversed anhedonia in two independent chronic stress models, suggesting that both mGlu2 and mGlu3 NAMs induce antidepressant-like effects through related but divergent mechanisms of action.

Anesthesia and Euthanasia of Brine Shrimp (Artemia franciscana).

Invertebrates are often overlooked as laboratory animals, yet they are commonly used in toxicology, developmental, cellular and molecular biology, and radiation studies with euthanasia as an endpoint. Little is known regarding appropriate euthanasia methods for invertebrate species, particularly for Artemia. Here, we evaluated the AVMA-recommended 2-step method of euthanasia in brine shrimp (Artemia franciscana). Artemia were exposed first to anesthetic solutions of 60% alcohol, 2.5 mg/L eugenol, or 4 g/L tricaine methanesulfonate (TMS) and then were transferred to euthanasia solutions of 70% alcohol, 95% alcohol, or 10% neutral buffered formalin. We examined time to anesthesia, behavioral response to anesthesia, anesthesia recovery, and time to euthanasia. Our results show that 2.5 mg/L eugenol and 4 g/L TMS inconsistently achieved anesthesia. Although 60% alcohol produced anesthesia, the time to anesthesia varied among replicate groups, and exposure resulted in an increase in abnormal behavior. We therefore do not recommend any of the tested anesthetic solutions for use in Artemia. Although all 3 euthanasia solutions were effective, more research is needed to provide recommendations regarding euthanasia for this species.

The Gain-of-Function Integrin ß3 Pro33 Variant Alters the Serotonin System in the Mouse Brain.

Engagement of integrins by the extracellular matrix initiates signaling cascades that drive a variety of cellular functions, including neuronal migration and axonal pathfinding in the brain. Multiple lines of evidence link the ITGB3 gene encoding the integrin ß3 subunit with the serotonin (5-HT) system, likely via its modulation of the 5-HT transporter (SERT). The ITGB3 coding polymorphism Leu33Pro (rs5918, PlA2) produces hyperactive αvß3 receptors that influence whole-blood 5-HT levels and may influence the risk for autism spectrum disorder (ASD). Using a phenome-wide scan of psychiatric diagnoses, we found significant, male-specific associations between the Pro33 allele and attention-deficit hyperactivity disorder and ASDs. Here, we used knock-in (KI) mice expressing an Itgb3 variant that phenocopies the human Pro33 variant to elucidate the consequences of constitutively enhanced αvß3 signaling to the 5-HT system in the brain. KI mice displayed deficits in multiple behaviors, including anxiety, repetitive, and social behaviors. Anatomical studies revealed a significant decrease in 5-HT synapses in the midbrain, accompanied by decreases in SERT activity and reduced localization of SERTs to integrin adhesion complexes in synapses of KI mice. Inhibition of focal adhesion kinase (FAK) rescued SERT function in synapses of KI mice, demonstrating that constitutive active FAK signaling downstream of the Pro32Pro33 integrin αvß3 suppresses SERT activity. Our studies identify a complex regulation of 5-HT homeostasis and behaviors by integrin αvß3, revealing an important role for integrins in modulating risk for neuropsychiatric disorders.SIGNIFICANCE STATEMENT The integrin ß3 Leu33Pro coding polymorphism has been associated with autism spectrum disorders (ASDs) within a subgroup of patients with elevated blood 5-HT levels, linking integrin ß3, 5-HT, and ASD risk. We capitalized on these interactions to demonstrate that the Pro33 coding variation in the murine integrin ß3 recapitulates the sex-dependent neurochemical and behavioral attributes of ASD. Using state-of-the-art techniques, we show that presynaptic 5-HT function is altered in these mice, and that the localization of 5-HT transporters to specific compartments within the synapse, disrupted by the integrin ß3 Pro33 mutation, is critical for appropriate reuptake of 5-HT. Our studies provide fundamental insight into the genetic network regulating 5-HT neurotransmission in the CNS that is also associated with ASD risk.

Loss of Serotonin Transporter Function Alters ADP-mediated Glycoprotein αIIbß3 Activation through Dysregulation of the 5-HT2A Receptor.

Reduced platelet aggregation and a mild bleeding phenotype have been observed in patients chronically taking selective serotonin reuptake inhibitors (SSRIs). However, it remains unclear how SSRIs, which inhibit the plasma membrane serotonin transporter (SERT), modulate hemostasis. Here, we examine how sustained inhibition of SERT activity alters serotonergic signaling and influences platelet activation and hemostasis. Pharmaceutical blockade (citalopram dosing) or genetic ablation (SERT(-/-)) of SERT function in vivo led to reduced serotonin (5-hydroxytryptamine (5-HT)) blood levels that paralleled a mild bleeding phenotype in mice. Transfusion of wild-type platelets to SERT(-/-) mice normalized bleeding times to wild-type levels, suggesting that loss of SERTs causes a deficiency in platelet activation. Although SERT(-/-) platelets displayed no difference in P-selectin or αIIbß3 activation upon stimulation with thrombin, ADP-mediated αIIbß3 activation is reduced in SERT(-/-) platelets. Additionally, synergistic potentiation of αIIbß3 activation by ADP and 5-HT is lost in SERT(-/-) platelets. Acute treatment of wild-type platelets with 5-HT2A receptor (5-HT2AR) antagonists or SSRIs revealed that functional 5-HT2ARs, not SERTs, are necessary for the synergistic activation of αIIbß3 by dual 5-HT/ADP stimulation. Pharmacological studies using radiolabeled guanosine 5'-3-O-([(35)S]thio)triphosphate and [(3)H]ketanserin revealed that platelets isolated from SERT(-/-) or citalopram-treated mice have reduced activation of G-proteins coupled to 5-HT2ARs and receptor surface expression. Taken together, these data demonstrate that sustained SERT loss of function reduces 5-HT2AR surface expression that is critical for the synergistic activation of αIIbß3 by 5-HT and ADP. These results highlight an antiplatelet strategy centered on blocking or desensitizing 5-HT2AR to attenuate ADP-mediated αIIbß3 activation.

Pro32Pro33 mutations in the integrin ß3 PSI domain result in αIIbß3 priming and enhanced adhesion: reversal of the hypercoagulability phenotype by the Src inhibitor SKI-606.

The plasma-membrane integrin αIIbß3 (CD41/CD61, GPIIbIIIa) is a major functional receptor in platelets during clotting. A common isoform of integrin ß3, Leu33Pro is associated with enhanced platelet function and increased risk for coronary thrombosis and stroke, although these findings remain controversial. To better understand the molecular mechanisms by which this sequence variation modifies platelet function, we produced transgenic knockin mice expressing a Pro32Pro33 integrin ß3. Consistent with reports utilizing human platelets, we found significantly reduced bleeding and clotting times, as well as increased in vivo thrombosis, in Pro32Pro33 homozygous mice. These alterations paralleled increases in platelet attachment and spreading onto fibrinogen resulting from enhanced integrin αIIbß3 function. Activation with protease-activated receptor 4- activating peptide, the main thrombin signaling receptor in mice, showed no significant difference in activation of Pro32Pro33 mice as compared with controls, suggesting that inside-out signaling remains intact. However, under unstimulated conditions, the Pro32Pro33 mutation led to elevated Src phosphorylation, facilitated by increased talin interactions with the ß3 cytoplasmic domain, indicating that the αIIbß3 intracellular domains are primed for activation while the ligand-binding domain remains unchanged. Acute dosing of animals with a Src inhibitor was sufficient to rescue the clotting phenotype in knockin mice to wild-type levels. Together, our data establish that the Pro32Pro33 structural alteration modifies the function of integrin αIIbß3, priming the integrin for outside-in signaling, ultimately leading to hypercoagulability. Furthermore, our data may support a novel approach to antiplatelet therapy by Src inhibition where hemostasis is maintained while reducing risk for cardiovascular disease.