Cdk8 Kinase Module Modifies Expression of Specific Translation-Related Proteins Before and After Stress

Translation is tightly coupled to growth status. Efficient protein synthesis is necessary for cell growth in nutrient rich environments, while global translation inhibition combined with selective translation of stress-responsive mRNAs helps limit growth in times of stress. Environmental stress cues which inhibit the nutrient-sensing complex TORC1 are known to reduce general translation, but how does the cell alter protein synthesis machinery to adapt to these conditions? A few mechanisms to promote cell survival in nitrogen starvation include post-translational modification and selective degradation of specific mRNA-binding translation factors, as well as inhibition of activators of genes whose products are required for general translation. How and when these occur, however, have remained elusive. Here, we demonstrate in Saccharomyces cerevisiae that the highly conserved Cdk8 kinase module (CKM) of the mediator complex (cyclin C, Cdk8, Med13, and Med12) transcriptionally upregulates specific 60S ribosome proteins and translation initiation factors such as eIF4G1 to maintain steady state levels of translation-related proteins in physiological conditions. Yeast CKM is known to predominantly repress stress response genes (SRG), and our previous findings revealed that SRG suppression is relieved through the degradation of Med13 and cyclin C following both cell survival and death cues. Our recent data further suggest that degradation of the CKM following nitrogen starvation also plays a transcriptional role in fine-tuning the expression of translation-related proteins. The CKM is thus a multi-faceted hub that can provide insight to how the cell adapts to stress at the levels of transcription, translation, and degradation

The Involvement of Ubiquitin in Med13 Cyclin C Degradation Following Cellular Stress

The Cdk8 Kinase Module is a dissociable regulator of cellular stress response genes, with degradation of its components Med13 and cyclin C eventually determining cell fate decisions such as engaging cell survival or cell death mechanisms. We aimed to explore the roles of ubiquitin in degradation of the Cdk8 Kinase Module following nitrogen starvation, with respect to the potential involvement of deubiquitinating enzyme Doa4, lysine linkage at position K63, and E2 ubiquitin conjugating enzymes Ubc4 and Ubc5. We utilized Western blot analysis to observe nitrogen starvation-induced degradation of Med13-HA in wild-type, doa4 mutant, and K63R yeast strains; degradation of cyclin C-MYC in wild-type and K63R strains; and Atg8-GFP activity in wild-type and ubc4/5 mutant strains, with Pgk1-GFP as a loading control. Results indicated that Med13 was degraded in doa4 mutant and K63R yeast strains, while cyclin C was stabilized in K63R strains. Additionally, GFP was cleaved from Atg8 in both wild-type and ubc4/5 mutant strains. We therefore determined that Med13 degradation does not require Doa4 or K63, and that Ubc4 and Ubc5 are specific to Med13 autophagy. However, K63 ubiquitin linkage is implicated in cyclin C degradation following nitrogen starvation, warranting further investigation

“Oscillating Between Hope and Despair”: A Narrative Case Study of Culture and Coping for Women in Engineering in Higher Education and Industry

Women in engineering in all phases (students, faculty, and industry) are traditionally underrepresented, and have been underrepresented in the field for decades (Eaton et al., 2020). The United States government has invested in STEM disciplines to address the low presence of women in STEM fields and the STEM workforce (National Academy of Sciences, 2016). Lower representation can be attributed to numerous factors, including a lack of institutional commitment, lack of representation throughout students’ upbringing, inappropriate cultural recruitment/outreach efforts, educational discrepancies throughout PK-12, and social expectations among others (Seymour & Hewitt 1997; Geisinger & Raman, 2013; Camacho et al., 2010; Smith et al., 2012). Not only is there low representation in the field of engineering, but also low retention for students, faculty, and industry due to the same factors that impact representation. The current qualitative study is a narrative case study utilizing relational interviewing. Participants included three women in the engineering field-- an undergraduate student, a professional engineer, and a faculty member and administrator. The key themes we uncovered-- Impact of Dominant Culture; Lack of Belonging/Connection; Justification of Existence; Emotional Turmoil; Coping Strategies—are woven throughout and across the narratives. Actionable change that we hope will come from this narrative study include determining ways we can make the engineering field more inclusive at all levels– in the classroom, on co-op, at universities, in academia, and on the field in industry

Training terminology on perceptions of women who engage in muscle strengthening activities

The present study examined the impact of muscle strengthening terminology on impressions formed of female exercisers, in addition to the influence of participant impression motivation and BSRI category on ratings of personality and physical attributes. Male and female participants ( N = 265, M age = 21.23) were presented with one of four vignettes describing a female target (weight trainer, resistance trainer, strength trainer, control). Participants then rated the target on personality and physical characteristics. Results indicated no significant differences among ratings of target types ( p \u3e .05). Moreover, the participants\u27 impression motivation did not influence target ratings ( p \u3e .05). A significant main effect emerged for BSRI category ( p \u3c .05). Participants classified as masculine-typed rated all targets as less kind compared to participants classified as feminine-typed or androgynous. It is possible the vignettes did not provide enough information about muscle strengthening to elicit stereotypes. Avenues for future directions are discussed

Membrane Potentials, Synaptic Responses, Neuronal Circuitry, Neuromodulation and Muscle Histology Using the Crayfish: Student Laboratory Exercises

The purpose of this report is to help develop an understanding of the effects caused by ion gradients across a biological membrane. Two aspects that influence a cell\u27s membrane potential and which we address in these experiments are: (1) Ion concentration of K+ on the outside of the membrane, and (2) the permeability of the membrane to specific ions. The crayfish abdominal extensor muscles are in groupings with some being tonic (slow) and others phasic (fast) in their biochemical and physiological phenotypes, as well as in their structure; the motor neurons that innervate these muscles are correspondingly different in functional characteristics. We use these muscles as well as the superficial, tonic abdominal flexor muscle to demonstrate properties in synaptic transmission. In addition, we introduce a sensory-CNS-motor neuron-muscle circuit to demonstrate the effect of cuticular sensory stimulation as well as the influence of neuromodulators on certain aspects of the circuit. With the techniques obtained in this exercise, one can begin to answer many questions remaining in other experimental preparations as well as in physiological applications related to medicine and health. We have demonstrated the usefulness of model invertebrate preparations to address fundamental questions pertinent to all animals

Building for the Exchange

Our project, “Building for the Exchange,” is trying to solve a problem that has been progressing for nearly a century. Modern development practices have changed the small business landscape in Morehead. It was once a vibrant downtown core with neighborhoods built around it. A surge in population over the last 70 years has caused businesses to spread across the community. The downtown has been left with empty storefronts, struggling to keep viable businesses. To try to solve this problem, our class was split into four groups and we were tasked to create business plans to fill vacant spaces in the downtown area of Morehead.https://scholarworks.moreheadstate.edu/celebration_posters_2023/1055/thumbnail.jp

Bentho-Pelagic Divergence of Cichlid Feeding Architecture Was Prodigious and Consistent during Multiple Adaptive Radiations within African Rift-Lakes

<b>Background</b> How particular changes in functional morphology can repeatedly promote ecological diversification is an active area of evolutionary investigation. The African rift-lake cichlids offer a calibrated time series of the most dramatic adaptive radiations of vertebrate trophic morphology yet described, and the replicate nature of these events provides a unique opportunity to test whether common changes in functional morphology have repeatedly facilitated their ecological success.<p></p> <b>Methodology/Principal Findings</b> Specimens from 87 genera of cichlid fishes endemic to Lakes Tanganyka, Malawi and Victoria were dissected in order to examine the functional morphology of cichlid feeding. We quantified shape using geometric morphometrics and compared patterns of morphological diversity using a series of analytical tests. The primary axes of divergence were conserved among all three radiations, and the most prevalent changes involved the size of the preorbital region of the skull. Even the fishes from the youngest of these lakes (Victoria), which exhibit the lowest amount of skull shape disparity, have undergone extensive preorbital evolution relative to other craniofacial traits. Such changes have large effects on feeding biomechanics, and can promote expansion into a wide array of niches along a bentho-pelagic ecomorphological axis.<p></p> <b>Conclusions/Significance</b> Here we show that specific changes in trophic anatomy have evolved repeatedly in the African rift lakes, and our results suggest that simple morphological alterations that have large ecological consequences are likely to constitute critical components of adaptive radiations in functional morphology. Such shifts may precede more complex shape changes as lineages diversify into unoccupied niches. The data presented here, combined with observations of other fish lineages, suggest that the preorbital region represents an evolutionary module that can respond quickly to natural selection when fishes colonize new lakes. Characterizing the changes in cichlid trophic morphology that have contributed to their extraordinary adaptive radiations has broad evolutionary implications, and such studies are necessary for directing future investigations into the proximate mechanisms that have shaped these spectacular phenomena

MPH Capstone experiences: promising practices and lessons learned

To ensure workforce readiness, graduate-level public health training programs must prepare students to collaborate with communities on improving public health practice and tools. The Council on Education for Public Health (CEPH) requires Master of Public Health (MPH) students to complete an Integrative Learning Experience (ILE) at the end of their program of study that yields a high-quality written product demonstrating synthesis of competencies. CEPH suggests written products ideally be “developed and delivered in a manner that is useful to external stakeholders, such as non-profit or governmental organizations.” However, there are limited examples of the ILE pedagogies and practices most likely to yield mutual benefit for students and community partners. To address this gap, we describe a community-led, year-long, group-based ILE for MPH students, called Capstone. This service-learning course aims to (1) increase capacity of students and partner organizations to address public health issues and promote health equity; (2) create new or improved public health resources, programs, services, and policies that promote health equity; (3) enhance student preparedness and marketability for careers in public health; and (4) strengthen campus-community partnerships. Since 2009, 127 Capstone teams affiliated with the Department of Health Behavior at the Gillings School of Global Public Health at The University of North Carolina at Chapel Hill have worked with seventy-nine partner organizations to provide over 103,000 h of in-kind service and produce 635 unique products or “deliverables.” This paper describes key promising practices of Capstone, specifically its staffing model; approach to project recruitment, selection, and matching; course format; and assignments. Using course evaluation data, we summarize student and community partner outcomes. Next, we share lessons learned from 13 years of program implementation and future directions for continuing to maximize student and community partner benefits. Finally, we provide recommendations for other programs interested in replicating the Capstone model

Environmental health conditions in protracted displacement: a systematic scoping review

Adequate environmental health services are critical for human rights, health, and development, especially in the context of forced displacement. There are more than 70 million forcibly displaced persons worldwide, most in protracted situations, having been displaced for more than two years. Some live in camps or informal settlements, but most live in urban areas. Environmental health services are important in the transition from emergency re- sponse to sustainable development in these settings, but evidence on environmental health in displaced popula- tions is disparate and of variable quality. We conducted a systematic scoping review of environmental conditions, exposures, and outcomes in protracted displacement settings; obstacles to improvement in environmental health services; and recommendations made for improvement. We included 213 publications from peer- reviewed and grey literature databases. Data were extracted on environmental health topics including water, sanitation, hygiene, overcrowding, waste management, energy supply, vector control, menstrual hygiene, air quality, and food safety. Most studies present data from low- and lower-middle income countries. Northern Africa and Western Asia and Sub-Saharan Africa are the most-represented regions. There is substantial evidence on water, sanitation, and crowding, but few studies report findings on other environmental health topics. Water-related disease, parasites, and respiratory infections are frequently cited and studies report that services often fail to meet international standards for humanitarian response. The most frequent obstacles and recommendations are institutional, political, or implementation-related, but few studies provide concrete recommendations for im- provement. Our review compiles and characterizes the research on environmental health in protracted displace- ment. We recommend including displaced populations in international environmental health policy and monitoring initiatives, and bridging from humanitarian response to sustainable development by preparing for long-term displacement from the early stages of a crisis

Ruthenium-cyclopentadienyl bipyridine-biotin based compounds: Synthesis and biological effect

Prospective anticancer metallodrugs should consider target-specific components in their design in order to overcome the limitations of the current chemotherapeutics. The inclusion of vitamins, which receptors are overexpressed in many cancer cell lines, has proven to be a valid strategy. Therefore, in this paper we report the synthesis and characterization of a set of new compounds [Ru(eta(5)-C5H5)(P(C6H4R)(3))(4,4'-R'-2,2'-bpy)](+) (R = F and R' = H, 3; R = F and R' = biotin, 4; R = OCH3 and R' = H, 5; R = OCH3 and R' = biotin, 6), inspired by the exceptional good results recently obtained for the analogue bearing a triphenylphosphane ligand. The precursors for these syntheses were also described following modified literature procedures, [Ru(eta(5)-C5H3)(P(C6H4R)(3))(2)Cl], where R is -F (1) or -OCH3 (2). The structure of all compounds is fully supported by spectroscopic and analytical techniques and by X-ray diffraction studies for compounds 2, 3, and 5. All cationic compounds are cytotoxic in the two breast cancer cell lines tested, MCF7 and MDA-MB-231, and much better than cisplatin under the same experimental conditions. The cytotoxicity of the biotinylated compounds seems to be related with the Ru uptake by the cells expressing biotin receptors, indicating a potential mediated uptake. Indeed, a biotin-avidin study confirmed that the attachment of biotin to the organometallic fragment still allows biotin recognition by the protein. Therefore, the biotinylated compounds might be potent anticancer drugs as they show cytotoxic effect in breast cancer cells at low dose dependent on the compounds' uptake, induce cell death by apoptosis and inhibit the colony formation of cancer cells causing also less severe side effects in zebrafish.This work was financed by the Portuguese Foundation for Science and Technology (Fundacao para a Crencia e Tecnologia, FCT) within the scope of Projects UID/QUI/00100/2019 and PTDC/QUI-QIN/28662/2017. This work was supported by the strategic program UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through the COMPETE2020 -Programa Operacional Competitividade e Intemacionalizacao (POCI). A.V. acknowledges the Investigator FCT2013 Initiative for the Project IF/01302/2013 and CEEC-IND/01974/2017 (acknowledging FCT, as well as POPH and FSE, the European Social Fund). L.C.-R, A.R.B. and A.P. thank FCT for their Ph.D. Grants (SFRH/BD/100515/2014, SFRH/BD/139271/2018, and SFRH/BD/139412/2018, respectively). L.C.-R also acknowledges Fulbright Research Grant 2017/2018 with the support of FCT. Brittany Karas acknowledges NJAES-RutgersNJ01201 and NIEHS Training Grant T32-ES 007148 and B.T.B. and C.D. acknowledge NIH-NIEHS P30 ES005022. K.R.C. acknowledges NJAES Project 01202 (W2045) and NIH ES005022