Freed–Hardeman University

Binding between DNA topoisomerase II and plasmid DNA can be measured using a variety of methods. The electrophoretic mobility shift assay (EMSA) is an efficient and inexpensive means for examining DNA affinity. This method relies on the principle that increasing concentrations of enzyme will result in more topoisomerase bound to DNA. As more binding occurs, the DNA migration in an agarose gel is decreased causing an upward "shift" in the DNA molecules when visualized in the gel. This shift can be compared across a range of concentrations to establish binding parameters under varying circumstances such as substrate type, the presence of a compound or drug, incubation time, or other conditions.

DNA Topoisomerase IIα (Top2A) is a nuclear enzyme that is a cancer drug target, and there is interest in identifying novel sites on the enzyme to inhibit cancer cells more selectively and to reduce off-target toxicity. The C-terminal domain (CTD) is one potential target, but it is an intrinsically disordered domain, which prevents structural analysis. Therefore, we set out to analyze the sequence of Top2A from 105 species using bioinformatic analysis, including the PSICalc algorithm, Shannon entropy analysis, and other approaches. Our results demonstrate that large (10th-order) interdependent clusters are found including non-proximal positions across the major domains of Top2A. Further, CTD-specific clusters of the third, fourth, and fifth order, including positions that had been previously analyzed via mutation and biochemical assays, were identified. Some of these clusters coincided with positions that, when mutated, either increased or decreased relaxation activity. Finally, sites of low Shannon entropy (i.e., low variation in amino acids at a given site) were identified and mapped as key positions in the CTD. Included in the low-entropy sites are phosphorylation sites and charged positions. Together, these results help to build a clearer picture of the critical positions in the CTD and provide potential sites/regions for further analysis.

Aim: This study aimed to explore the clinic learning models through student perspectives using that insight to explore how these students viewed a newly established speech clinic program and understand the organizational and learning benefits and limitations of the clinic and how it can be improved for future service for student learning.
 Methods: This exploratory study utilized an observation analysis to understand how the student-led functioning of the clinic fits into both the learning models of Bloom’s Taxonomy and Bandura’s Triadic Reciprocal Model of Causality and the factors that contribute most to these learning models. These models were chosen due to the systemic and progressive nature of learning that reflects the academic mission of the university’s accrediting body: the Southern Association of Colleges and Schools Commission on Colleges. A mixed closed and open-ended student questionnaire was utilized to gain insight into themes that influence the student-led learning model.
 Results: The clinic was overall seen as a place of relationship building and skills development. The main contributors to this relationship-building and skills development lie in the ability of students to apply what they have been learning in the classroom to a functional situation, as well as the opportunity to serve the community and build confidence and leadership skills.
 Conclusion: Student leadership roles in the clinic are leading to self-efficacy which is seen in the highest level of Bloom’s Taxonomy and Bandura’s Triadic Reciprocal Model of Causality.

The intersection of religion and science often elicits polarizing views among scientists, though approximately half of American scientists identify as religious. Mounting evidence also supports the role of spirituality in comprehensive patient care. The purpose of this study was to explore the religiosity of faculty who teach in the anatomical sciences at U.S. colleges and universities. Surveys were administered to anatomists through two professional societies. Two-thirds (64.9%, 74/114) of respondents identified as religious, 26.3% (30/114) as atheist, and 8.8% (10/114) as agnostic. Most respondents (64.9%, 74/114) disagreed with the statement, "There is no place for religion and science to intersect." Approximately one in three respondents expressed concern that sharing/disclosing their religious beliefs would negatively affect the perceptions of colleagues (32.5%, 37/114) and students (28.9%, 33/114) toward them. Faculty at faith-based institutions were more open to disclosing their beliefs (p = 0.045), and highly religious individuals were more concerned (p = 0.001). Fewer than one-fifth of respondents 17.5% (20/114) personally experienced mistreatment or discrimination within academic settings due to their religious beliefs. Most respondents held politically liberal-leaning views (71.0%, 76/107). Highly religious individuals were more likely to be politically conservative (p < 0.001). Overall, this study demonstrates that the number of anatomists who identify as religious may be higher than that of other biological disciplines and that mistreatment due to religious views remains a challenge for some in the profession. Continued dialogue regarding the role of religion in professional identity expression may be an important step in mitigating religion-focused mistreatment and discrimination in academic settings.

Abstract Idiopathic pulmonary fibrosis (IPF) is an aggressive interstitial lung disease associated with progressive and irreversible deterioration of respiratory functions that lacks curative therapies. Despite IPF being associated with a dysregulated immune response, current antifibrotics aim only at limiting fibroproliferation. We show here that the P2RX7/IL-18/IFNG axis is downregulated in IPF patients and that P2RX7 has immunoregulatory functions. Using our positive modulator of P2RX7, we show that activation of the P2RX7/IL-18 axis in immune cells limits lung fibrosis progression in a mouse model by favoring an anti-fibrotic immune environment, with notably an enhanced IL-18-dependent IFN-γ production by lung T cells leading to a decreased production of IL-17 and TGFβ. Overall, we show the ability of the immune system to limit lung fibrosis progression by targeting the immunomodulator P2RX7. Hence, treatment with a small activator of P2RX7 may represent a promising strategy to help patients with lung fibrosis.

Sometimes, despite the most perfectly delivered writing lesson implemented with fidelity, students with emotional and behavioral disorders (EBD) do not make the progress expected by teachers or themselves. In these instances, teachers may individualize and differentiate for students with EBD not making adequate progress by purposefully intensifying elements of the writing instruction and supports provided to students. This article describes and explains how to intensify writing instruction to meet student needs.

Gap-enhanced Raman tags are a new type of optical probe that have wide applications in sensing and detection. A gap-enhanced Raman tag is prepared by embedding Raman molecules inside a gap between two plasmonic metals such as an Au core and Au shell. Even though placing Raman molecules beneath an Au shell seems counter-intuitive, it has been shown that such systems produce a stronger surface-enhanced Raman scattering response due to the strong electric field inside the gap. While the theoretical support of the stronger electric field inside the gap was provided in the literature, a comprehensive understanding of how the electric field inside the gap compares with that of the outer surface of the particle was not readily available. We investigated Au@SiO2@Au nanoparticles with diameters ranging from 35 nm to 70 nm with varying shell (2.5-10 nm) and gap (2.5-15 nm) thicknesses and obtained both far-field and near-field spectra. The extinction spectra from these particles always have two peaks. The low-energy peak redshifts with the decreasing shell thickness. However, when the gap thickness decreases, the low-energy peaks first blueshift and then redshift, producing a C-shape in the peak position. For every system we investigated, the near-field enhancement spectra were stronger inside the gap than on the outer surface of the nanoparticle. We find that a thin shell combined with a thin gap will produce the greatest near-field enhancement inside the gap. Our work fills the knowledge gap between the exciting potential applications of gap-enhanced Raman tags and the fundamental knowledge of enhancement provided by the gap.

Abstract Bone and joint infections (BJIs) are difficult to treat and affect a growing number of patients, in which relapses are observed in 10-20% of the case. These relapses, which call for prolonged antibiotic treatment and increase the risk of emergence of resistance, may originate from ill understood adaptation of the pathogen to the host. Here, we studied three pairs ofEscherichia colistrains corresponding to three cases of BJIs and their relapse to better understand in-patient adaptation.Whole genome comparison presented evidence for positive selection with prevalence of non-synonymous and loss of function mutations. Phenotypic characterization showed that biofilm formation capacity was not modified, contrary to what is usually described in such relapse cases. Although virulence was not modified, we identified the loss of two virulence factors (namely an AFA afimbrial adhesin and a YadA-like adhesin) contributing to immune system evasion in one of the studied relapse strain. Other selected strategies likely helped the relapse strains to outcompete competitors through global growth optimization and colicin production. This work highlights the variety of strategies allowing in-patient adaptation in BJIs.

The purpose of this study was to investigate a set of teacher-level factors that educator preparation providers (EPPs) may be examining to anticipate how effective teachers might be in the classroom. An ordinal logistic regression was performed to determine impact on teacher effectiveness, given five predictor variables: level (undergraduate vs. graduate), age, GPA, ACT, and Praxis II scores. The findings demonstrated that only level made a significant individual contribution to prediction, suggesting that undergraduate students were less likely to have higher teacher effectiveness scores than graduate students. The major implication for states may be to evaluate EPPs and their abilities to impact the quality of preservice teacher candidates separately based on whether they serve primarily undergraduate or graduate student populations.

This article discusses the similarities between the biographical presentations of Philo and Origen in Eusebius' Ecclesiastical History. The tension Eusebius feels between Philo Christianus and Philo Judaeus is certainly detectible in his presentation of the Therapeutae, a group about whom Philo reports and whom Eusebius considers the first Egyptian Christians. Eusebius recognizes that Philo is exegetically closer to Christianity, and religiously closer to Judaism. This realization creates an ambiguity in the Ecclesiastical History in which Philo is presented explicitly neither as Jew nor Christian, but can be identified as either.