Theory to Practice: Integrating Service-Learning into a Pre-Practicum Introduction to School Counseling Course

Service learning is a method of instruction that integrates volunteerism, as well as reflection, in the classroom. The purpose of this article is, therefore, to describe the implementation and design of a pre-practicum service learning experience into an Introduction to School Counseling course. The authors describe the empirically grounded design of the service learning experience, as well as the course goals, how it was integrated into the class, and a summary of student experiences

Exploring Pre-practicum Site-Based Experiential Learning in School Counselor Preparation

In this study, participation in a pre-practicum site-based experiential learning assignment was examined. First semester pre-service school counselors who were enrolled in an introductory school counseling course, engaged in observation, counseling, and academic support with students and clients at community and school sites. Qualitative data was collected across three cohort groups and examined through phenomenological inquiry. Participants shared common themes, including that they: (a) Recognized inequities while they expanded their worldview; (b) Preferred a structured experience; (c) Valued relationships; and (d) Drew connections to the classroom and the field

The Effects of Rhodiola Crenulata Extract on Proliferation and Differentiation in Glioblastoma Multiforme

Purpose: Purpose of the study was to evaluate the effects of rhodiola crenulata plant extract on glioblastoma in vitro. Methods: U-87MG glioblastoma multiforme cell line was utilized for evaluation in this study. Cells were treated with 100ug/ml or 200ug/ml of rhodiola crenulata and compared to ethanol vehicle control. Proliferation was measured at 24, 48, 72, and 96 hours after treatment utilizing an MTS proliferation assay. To further assess proliferation a clonogenicity assay was conducted. These cells were treated with ethanol vehicle control, 100ug/ml of rhodiola, radiation, or combined rhodiola/radiation treatment. To evaluate differentiation the expression of glial fibrillary acidic protein (GFAP), a protein marker of differentiation, was assessed with immunocytochemistry. Results: Effects on proliferation were initially noted at 48hours after treatment and observed through the 96-hour period. The effects on proliferation were noted in both treatment groups. At 96-hours after treatment significant difference was noted between the 100ug/ml of rhodiola and control group (p=0.0065) and significant difference noted between the 200ug/ml of rhodiola and control group (p=0.0006). Cell clonogenicity was reduced in the cells treated with 100ug/ml of rhodiola. The decreased number of colonies was significant when comparing the radiation treated cells with 100ug/ml rhodiola treated cells (p=0.0030). GFAP was overexpressed in the rhodiola treatment group when compared to expression in the control group (Figure 1). Conclusion: Rhodiola crenulata extract effectively decreases proliferation and increases differentiation of glioblastoma cells in vitro. Further work is required to fully understand the extent and full effects rhodiola crenulata has glioblastoma cells

A Novel Approach to Targeted Oncologic Therapy - Co-culture Viability of Polymer Prodrug Conjugation to Mesenchymal Stem Cells

Background/Purpose: Conjugation of polymer prodrugs to tumor homing cells, such as Mesenchymal Stem Cells (MSCs), could provide a vehicle for actively targeted delivery of polymer prodrugs. Methods: Human Bone Marrow MSCs were conjugated to either a doxorubicin polymer prodrug or free doxorubicin and were co-cultured with T-cells. Viability was assessed through the use of a Vi-cell counter. In Vivo Migration Analysis was performed NOD SCID mice implanted with subcutaneous MDA MB-231 breast cancer xenografts. Following tumor establishment, mice were injected via lateral tail vein injection with either saline or polymer loaded MSCs. Five days following stem cell injection, mice were euthanized, tumors were harvested and sections were analyzed using fluorescent microscopy and immuno-histochemical staining for cd105. Results: T-cell viability was reduced when co-cultured with MSCs conjugated to free doxorubicin although cells co-cultured with MSCs conjugated to doxorubicin polymer did not exhibit reduced viability. Polymer loaded MSCs displayed intact tumor homing migratory ability in vivo (Figure 1). Conclusion: MSCs conjugated to doxorubicin released the drug, resulting in reduced neighboring T-cells viability. MSCs loaded with polymer maintained their migratory capacity were able to migrate to tumors in vivo. MSCs therefore represent a potential vehicle for targeted drug delivery. Future work will focus on developing methods for releasing the drug upon successful delivery to the target in vivo

In Vivo Evaluation of a Biomimetic Polymer-Doxorubicin Conjugate for Cancer Therapy

This poster will describe a novel polymer pro-drug platform designed for conjugation and delivery of chemotherapeutics. Specifically, polymer pro-drugs were prepared from functional polymer zwitterions and doxorubicin (DOX), and evaluated in vivo to assess toxicological, pharmacokinetic and therapeutic properties. The biocompatible polymer scaffold (PolyMPC) consists of zwitterionic phosphorylcholine pendent groups, which mimic the natural hydrophilic moieties of phospholipids in cell membranes, and hydrazone linkages that allow for pH-triggered release of DOX. PolyMPC-DOX pro-drugs were isolated as dry solids using a facile strategy that allows for precise control of molecular weight and DOX incorporation. In vivo toxicity of PolyMPC and PolyMPC-DOX was assessed in a murine model. The maximum tolerated dose of the pro-drug was five times greater than that of free DOX, while PolyMPC alone exhibited no toxicity even at a dose of 800 mg/kg. A pharmacokinetic study in tumor-bearing mice demonstrated a significant increase in circulation half-life of conjugated DOX (t1/2=2 hours) compared to free DOX (t1/2=15 minutes), with conjugated DOX detectable in blood serum for longer than 24 hours. This pronounced enhancement in circulation time was attributed to the macromolecular scaffold, which precludes rapid renal clearance compared to native DOX. Examination of mice given PolyMPC-DOX five days after injection in the PK study showed a three-fold increase of drug accumulated in tumor tissue compared to that of mice treated with free DOX and drug accumulation in off-target organs was reduced for mice given DOX conjugate. The therapeutic efficacy of the PolyMPC-DOX conjugates was then assessed in an orthotopic murine breast cancer model. The treatment group given PolyMPC-DOX exhibited a two-fold increase in overall survival and a significant reduction in average tumor volume compared to the free DOX and saline control groups. A study evaluating the therapeutic efficacy of PolyMPC-DOX in a human ovarian xenograft tumor model is ongoing

Identification of nicotine-seeking and avoiding larval zebrafish using a new three-choice behavioral assay

IntroductionNicotine dependence is one of the main causes of preventable diseases in the United States. Nicotine-seeking and avoidance behavioral assays in larval zebrafish could be used for identifying potential new pharmacotherapeutics in an early phase of drug discovery and could facilitate the identification of genes and genomic variations associated with nicotine-seeking and avoidance behavior.MethodsA new three-choice behavioral assay has been developed for the identification of nicotine-seeking and avoiding larval zebrafish. The three choices are represented by three compartments of a gradient maze. Video-recording and subsequent quantitative analysis of the swimming track was carried out using EthovisionXT (Noldus).ResultsThree behavioral phenotypes could be identified. Nicotine-seeking larval zebrafish occupied nicotine compartments for longer periods and entered the nicotine-containing compartments most frequently. Nicotine-avoiders spent most of the cumulative time in the water compartment or entered the water compartment most frequently. Non-seekers remained in the center compartment for most of the time. In the gradient maze, about 20–30% of larval zebrafish had a preference for low nicotine concentrations whereas nicotine avoidance was stronger at higher nicotine concentrations. Lower concentrations of nicotine (0.63 μM, 6.3 μM) resulted in higher percentages of nicotine seekers whereas high nicotine concentrations (63 μM, 630 µM) resulted in higher percentages of nicotine avoiders. Pre-treatment of larval zebrafish with nicotine slightly increased the percentage of nicotine avoiders at lower nicotine concentrations. Treatment with varenicline strongly increased the percentage of nicotine avoiders at lower nicotine concentrations.ConclusionThe results show that larval zebrafish have individual preferences for nicotine that could change with drug treatment. The three-choice gradient maze assay for larval zebrafish provides a new testing paradigm for studying the molecular and cellular mechanisms of nicotine action and the discovery of potential new pharmacotherapeutics for the treatment of smoking cessation

Acute multi-sgRNA knockdown of KEOPS complex genes reproduces the microcephaly phenotype of the stable knockout zebrafish model

Until recently, morpholino oligonucleotides have been widely employed in zebrafish as an acute and efficient loss-of-function assay. However, off-target effects and reproducibility issues when compared to stable knockout lines have compromised their further use. Here we employed an acute CRISPR/Cas approach using multiple single guide RNAs targeting simultaneously different positions in two exemplar genes (osgep or tprkb) to increase the likelihood of generating mutations on both alleles in the injected F0 generation and to achieve a similar effect as morpholinos but with the reproducibility of stable lines. This multi single guide RNA approach resulted in median likelihoods for at least one mutation on each allele of >99% and sgRNA specific insertion/deletion profiles as revealed by deep-sequencing. Immunoblot showed a significant reduction for Osgep and Tprkb proteins. For both genes, the acute multi-sgRNA knockout recapitulated the microcephaly phenotype and reduction in survival that we observed previously in stable knockout lines, though milder in the acute multi-sgRNA knockout. Finally, we quantify the degree of mutagenesis by deep sequencing, and provide a mathematical model to quantitate the chance for a biallelic loss-of-function mutation. Our findings can be generalized to acute and stable CRISPR/Cas targeting for any zebrafish gene of interest

The Effect of 3D Printed Models on Surgical Planning and Outcomes for Partial Nephrectomies

Purpose: Partial nephrectomies to remove renal masses are kidney-sparing procedures that rely on anatomical comprehension. Prior research demonstrates complex and expensive 3D models (~$1000) increased surgeons’ confidence in selecting their operative plan. We aim to use simpler and cheaper 3D models to improve preoperative surgeon confidence and support operative management. Methods: 3D printed models (~$35) of the affected kidney, mass, and renal vasculature were created using preoperative CT or MRI imaging of 17 patients presenting for partial nephrectomies at Thomas Jefferson University (TJU) between July and December 2020. The models were created at TJU using Ultimaker technology. Surgeons filled out three surveys assessing their surgical plan and confidence in the plan: before seeing the model, after seeing the model before surgery, and after surgery. Patients with 3D modeling were crossmatched by demographics and operative technique with patients without 3D modeling who had partial nephrectomies between 2018 - 2019. Results and Conclusions: 16 of 17 attendings completed the surveys. Surgeon confidence increased before (7.6) and after seeing the 3D model (7.9) on a 10-point scale (10=most confident). On postoperative surveys, attendings rated the models 8.3 out of 10 in their helpfulness to anatomical comprehension. Patients with 3D modeling had slightly higher rates of selective renal artery clamping over complete vascular clamping when compared to crossmatched partial nephrectomies without 3D modeling. Cost-effective 3D models can be helpful tools for surgeons to understand anatomical relationships and reduce complete vascular clamping that may be difficult with imaging alone. Other surgical fields may benefit from preoperative education through 3D modeling

DU Undergraduate Showcase: Research, Scholarship, and Creative Works: Abstracts

Abstracts from the DU Undergraduate Showcase

Clinical Sequencing Exploratory Research Consortium: Accelerating Evidence-Based Practice of Genomic Medicine

Despite rapid technical progress and demonstrable effectiveness for some types of diagnosis and therapy, much remains to be learned about clinical genome and exome sequencing (CGES) and its role within the practice of medicine. The Clinical Sequencing Exploratory Research (CSER) consortium includes 18 extramural research projects, one National Human Genome Research Institute (NHGRI) intramural project, and a coordinating center funded by the NHGRI and National Cancer Institute. The consortium is exploring analytic and clinical validity and utility, as well as the ethical, legal, and social implications of sequencing via multidisciplinary approaches; it has thus far recruited 5,577 participants across a spectrum of symptomatic and healthy children and adults by utilizing both germline and cancer sequencing. The CSER consortium is analyzing data and creating publically available procedures and tools related to participant preferences and consent, variant classification, disclosure and management of primary and secondary findings, health outcomes, and integration with electronic health records. Future research directions will refine measures of clinical utility of CGES in both germline and somatic testing, evaluate the use of CGES for screening in healthy individuals, explore the penetrance of pathogenic variants through extensive phenotyping, reduce discordances in public databases of genes and variants, examine social and ethnic disparities in the provision of genomics services, explore regulatory issues, and estimate the value and downstream costs of sequencing. The CSER consortium has established a shared community of research sites by using diverse approaches to pursue the evidence-based development of best practices in genomic medicine