• Development And Characterization Of Polymeric Formulations And Microneedles For Dermal Drug Delivery

      Kim, Yujin
      Skin provides an attractive route for drug delivery, as transdermal delivery systems can ensure noninvasive and sustained delivery of medication across the skin into systemic circulation. Although the skin is a convenient site for drug administration, it only permits passive penetration of permeants with certain physicochemical properties. As a result, different strategies or devices have been developed to enhance the drug permeation into and across skin. The first aim of this study was to evaluate the efficacy of multiple dosing of trolamine salicylate (TS) as a topical analgesic on in vitro skin permeation. Also, the effect of sonophoresis on topical delivery of analgesic was evaluated. Results suggested that the multiple dosing (17.4 ± 4.1 μg/sq.cm) delivered a significantly higher amount of drug than the single dose (6.5 ± 0.6 μg/sq.cm). Also, the use of ultrasound enhances topical absorption of TS into and across skin. In the second aim, we focused on developing a new formulation to reduce the frequency of application of topical analgesics. A Poly lactic-co-glycolic acid (PLGA) based bio-adhesive polymeric solutions were prepared successfully and formed a thin film upon application in situ. A significantly higher amount of TS was delivered from a formulation containing 20% PLGA (45 ± 4 µg/sq.cm) as compared to PLGA-free counterpart (0.6 ± 0.2 µg/sq.cm). In the third aim, hyaluronic acid (HA) microneedles (MNs) were fabricated with and without magnesium ascorbyl phosphate (MAP) to facilitate the delivery of the active agent into and across the skin. The results showed that successful fabrication of HA MN, and the application of the current MN technology enhanced the delivery of MAP into skin (96.8±3.9 µg/sq.cm) compared to passive delivery (44.9±16.3 µg/sq.cm). In the fourth aim, the feasibility of transdermal delivery of heparin, a hydrophilic macromolecule, through laser-microporated skin was investigated. There was no passive permeation of heparin across the skin. However, permeation of heparin from the laser-treated group delivered 13.4 ± 0.62 µg/sq.cm after 24 h. Furthermore, heparin was not delivered to either the epidermis or dermis passively, whereas the laser-treatment group enabled delivery of heparin to the epidermis as well as the dermis.
    • Development And Evaluation Of An Adolescent Chronic Restraint Stress (acrs) Protocol To Model Adult Depression In Female Rats / By Meghan Hibicke.

      Hibicke, Meghan
      Major depressive disorder (MDD), a mood disorder that disproportionately affects women, is often predicated by childhood or adolescent stress. However, females have traditionally been excluded from pharmacological research, and the influence of adolescent stress on depressive-like behaviors has not been widely reported. Additionally, reports correlating adolescent stress with adult depressive-like behaviors have largely neglected to evaluate the effect of common antidepressants. The purpose of this research was to develop and validate an adolescent chronic restraint stress (aCRS) protocol using female rats in order to address the impact of adolescent stress on female adult depressive-like characteristics. During the pilot study, rats were restrained 60 minutes daily for 12 consecutive days beginning in early adolescence, then allowed to mature to adulthood. Control and restrained animals were handled and weighed daily. Behavioral testing included sucrose preference testing (SPT), elevated plus maze (EPM), locomotor activity, novel object recognition (NOR) and object location (OLT), and forced swim tests (FST). Rats were sacrificed immediately following the FST, and trunk blood was collected for corticosterone (CORT) analysis. Four subsequent studies modified the aCRS protocol by increasing the restraint period from 12 to 14 days, using exact-age animals, and by including a treatment period during which antidepressants (fluoxetine and desipramine) or saline were given, and by changing the number and timing of the behavioral assays and sample collections. Additionally, brain-derived neurotrophic factor (BDNF) was assessed. In all studies, aCRS rats displayed significantly increased depressive-like behavior in the FST, but no differences in anxiety-like behaviors or locomotor activity vs control rats. Desipramine, but not fluoxetine, consistently decreased depressive-like behavior in restrained rats. Results of SPT, NOR, OLT, and CORT were inconsistent across experiments. Serum, but not HIP or FCX BDNF, positively correlated with immobility in the FST (a depressive-like behavior), which is not consistent with the literature. In the final experiment, we observed that uterine weights were not normally distributed, with aCRS rats trending low. In conclusion, aCRS elicits increased depressive-like behaviors in adult female rats, without altering anxiety-like behaviors or locomotor activity. Future investigations using aCRS rats should include further evaluation of reproductive hormones and BDNF.
    • Development and Evaluation of Micro/Nanoparticulate Vaccines for Human Papillomavirus and Influenza

      Vo, Trinh; College of Pharmacy
      Influenza and Human papillomavirus (HPV) are among the most common infectious deseases prevalent globally. The influenza viruses cause approximately 200,000 hospitalizations each year, resulting in a significant financial burden to the healthcare industry. Similarly, HPVs cause virulent infections that have multiple serotypes capable of producing genital warts and cervical cancer. In both cases the infection is spread through viral agents, The influenza virus is transmitted by aerosolized mist of coughs and sneezes with HPV is a sexually transmitted disease. Prevention of influenza and HPV is possible with the help of vaccines, and currently, there are several vaccines for both infectious diseases. However, in the case of influenza, development of a vaccine is challenging as the virus undergoes rapid mutations and hence, the vaccine needs to be modified every season. Along with the constant need for vaccination rates. In the case of HPV, the need for a novel vaccine delivery system is more crucial. HPV is directly linked to cervical cancer and developing a novel strategy to improve vaccination globally is a major priority. Commercial vaccines are not only expensive but in developing countries the financial cost of maintaining cold storage and vaccine administration is significantly challenging. The developing non-parenteral vaccines is essential to improve patient compliance without affecting safety and efficacy of the vaccine. This project has two main sections focusing on development of an oral microparticulate vaccine for influenza and HPV. In the first part of this project, we developed an oral microparticulate vaccine for influenza. The influenza vaccine consists of a M2e protein which is a viral protein common to all influenza viruses encapsulated into a microparticle. Adjuvants such as Alum, MPL. R848, MF59, Flagellin, CpG and P4 were encapsulated in microparticles separately. Microphages werestimulated with the vaccine-adjuvants combinations and subsequently., splenocytes were incubated with activated macrophages to investigate the T-cell activity. We observed significant enhancement of immune markers such as nitric oxide, CD80, CD86, and MHC II with several adjuvants. Furthermore, CD4 and CD8 T cell expression was also significantly enhanced in groups treated with particulate adjuvants. In the second part of this project, we formulated a novel oral and transdermal vaccine delivery system for HPV. Development of potent vaccines against HPV can prevent serious complications such as genital warts and cervical cancer. Therefore a novel non-parenteral vaccine against HPV may significantly reduce incidence of HPV worldwide. In this study, three different microparticualte vaccine formulations have been prepared and characterized: Bovine Albumin Serum (BSA) cross0linked with glutaradehyde; b-cyclodextrin, and cellulose acetate succinate matrices. We systematically studied activation of the immune markers such as nitirc oxide. CD80, CD86, CD40 and MHC-II. In vivo studies demonstrated elevated humoral and cell-mediated immune response in murine models. We observed elevated IgG titers in mice vaccinated with microparticulate HPV16 VLP vaccine, Furthermore, flow cytometry analysis revealed expansion of specific T-cell populations in vivo. This study proves the efficacy of the HPV-16 oral microparticulate vaccine as a promising alternative to conventional vaccines, In the alter part of the HPV vaccine project we investigated the potential of transdermal HPV vaccine as a novel strategy to improve vaccinations globally. In order to combat HPV infection, vaccines must be affordable, self-administered and efficacious. Transdermal vaccines can be self-administered. are almost painless and tap the immune cells present under the skin. Transdermal vaccines have gained immense attention due to their efficacy. In this project, our main aim was to develop a microparticulate HPV16 VLP vaccine for transdermal vaccination. HPV-16 VLP was encapsulated in microparticles and characterized for size, zeta potential, encapsulation efficiency and microparticle yield. Scanning electron microscope images confirmed the particles were irregular with surface indentations. Western blot analysis was performed on HPV16 VLP extracted from microparticles, which confirmed the retention of antigenicity of the epitope following microparticles, which confirmed the retention of antigenicity of the epitope following microparticle fromulation. Following in vitro characterization studies, we performed extensive in vivo studies to investigate the potential of the microparticulate vaccine to initiate a robust and sustained immune response. IgC titers for mice vaccinated with microparticulate vaccine were significantly elevated as compared to antigen solutions adminstered by the transdermal route. Flow cytometry was performed on vaccinated and control groups to investigate the expansion of specific T-cell populations. Vaccinated mice demonstrated significant increase in CD4, CD45R, CD27 and CD62L cell populations. Expansion of these specific T-cell subsets demonstrates the efficacy of the microparticulate vaccine when administered transdermally. Thus, our study proves the efficacy of the microparticulate vaccine over vaccine solutions and the novel approach of administering the vaccine through a minimally invasive, pain-free route that will help to improve acceptability of vaccination against HPV.
    • Development of Microencapsulated Formulations for Prostate Cancer Vaccine and Live Cells

      Akalkotkar, Archana; College of Pharmacy
      In the current trend, novel particle based platform technology has paved its way to various therapeutic benefits in our lives. In this study, we explored the way of harnessing this technology in order to formulate effective vaccines in the quest for mass immunization in a cost-effective manner. Here we propose a custom formulation of vaccines, which will render effects of sustained release, site specificity and cost-effective delivery of various antigens to enhance the immunity. We used the novel nano-micro technology based of Spray drying as a platform to deliver vaccines against Prostate cancer and encapsulated live cells. In this study, we formulated a whole cell lysate vaccine against prostate cancer via oral and transdermal route of administration. The formulation intended for oral adminstration was optimized for M-cell targeting, The formulation increased the immunogenicity of the vaccine by incorporating the antigen into an albumin matrix having a size of around 0.35-1.2 um that acted as a synthetic adjuvant. The animals were vaccinated with 1 prime and 4 booster doses administered every 14 days over 10 weeks duration, followed by challenge with live tumor cells which showed protection after oral vaccination. Unlike subcutaneous injections, administration-using microneedles is painless and in general can increase the permeability of many compounds ranging in size from small molecules to proteins and microparticles that do not normally penetrate the skin. The mice vaccinated via transdermal route showed promising results in delaying the tumor growth compared to the controls. The study was extrapolated in a therapeutic approach. This would mimic the real clinical scenario where the patient who was diagnosed with prostate cancer would come in the clinic for immunotherapy. The microparticle based delivery system showed protection in vaccinated group compared to the controls. This shows the microparticle based delivery system can be an useful tool for delivery as well as live cells in vivo.
    • Development of Spray Dried Microparticulate Delivery Systems for Vaccines and Oligonucleotides

      Ubale, Ruhi V; College of Pharmacy
      In recent years, research related to the development of biotherapeutics has gained tremendous interest in industry as well as academia. There are a number of biologics available in the market and many more currently being evaluated in clinical trials. Their development has gained momentum due to the range of advantages they offer over traditional small molecule drugs. However, there still remain challenges with their delivery mainly due to the poor stability of these molecules in harsh physiological conditions. A number of strategies are being devised for the delivery of biomolecules by non-invasive routes to make delivery easier and more patient compliant. Microparticles are one of the options being explored as they offer protection, sustained release and enhanced bioactivity to the biomolecule. In this project, we have studied the use of microparticles for the development of formulations for a meningococcal polysaccharide vaccine and an antisense oligonucleotide to NF-kB. Meningitis occurs majorly in children and is caused by Neisseria meninigitidis. Infection by this bacterium spreads rapidly through the body resulting in loss of limbs, hearing and can also be fatal. Hence, vaccination is considered as the only means prevention for meninigitis. Marketed vaccines use protein conjugated bacterial capsular polysaccharides as the antigen. Polysacchatides are considered inherently weak antigens with a T- independent immune response. With the development of a microparticulate vaccine using a protien matrix, we expected to stimulate the T-dependent immune pathway for these antigens avoiding the use of a conjugated Diphtheria toxoid. We have studied the enhancement of innate immune responses by the microparticulate vaccine. The antigen used N. meningitidis polysaccharide A, aslo an adjuvant- kdtA(unglecosylated lipid A)- was studied for its response. The microparticulate vaccine was characterized for its physicochemical characteristics like size, charge and toxicity. Also it was studied for the innate immune responses the microparticles can elicit after incubation with murine and human macrophages. Release of innate immune markers like TNF-a, IL-1B, IL-8, nitric oxide and reactive oxygen species from macrophages was studied and the microparticulate vaccine was observed to elicit a desirable innate immune response. Antigen presentation by macropaphages stimulated with the microparticulate vaccine was also studied by visualizing autophagy. Lung inflammation occurs doe to an infection or a damaging agent that irritates the lung lining or pleura. NF-kB is a nuclear transcription factor activated by stimuli like endotoxin and bacteria that initiates the synthesis of pro inflammatory cytokines such as TNF-a, IL-1, IL-6, etc. Antisense are single stranded RNA complementary to a chosen sequence that offer great potential to inhibit the synthesis of individual proteins by interfering with protein translation. As intracellular penetration of antisence compounds has proven to be a limiting factor in their effectiveness, we have proposed to develop a microparticulate formulation for pulmonary delivery of antisense to NF-kB and evaluate their efficacy. The microparticulate formulation of antisense to NF-kB was characterized for its physiochemical characteristics like size, charge, antisense content and release. Since enhancing uptake into cells was the crux of the study, we also visualized and quantified uptake of microparticles by microphages. We looked at the period of lung resistance and the biodistribution of the microparticles using a near infrared bioimager. Serum levels of proinflammatory cytokines were studied after induction of lung inflammation in a rat model. Animals that were delivered antisense microparticles to the lung showed reduced levels of TNF-a and IL-1b.
    • Diffusion Of And Access To Innovation: The Efficacy Of Bring-your-own-device (byod) Programs To Improve Academic Achievement Of Students From Low-income Families

      Coston-Scott, Marcia Elizabeth
      ABSTRACT MARCIA COSTON-SCOTT THE EFFICACY OF BRING-YOUR-OWN-DEVICE (BYOD) PROGRAMS TO IMPROVE ACADEMIC ACHIEVEMENT OF STUDENTS FROM LOW-INCOME FAMILIES Under the direction of OLIVIA M. BOGGS, Ed.D. The purpose of this mini-ethnographic case study was to examine the perspectives of school leaders and teachers regarding the effectiveness of mobile learning devices in improving academic achievement of low-income students in the disciplines of mathematics and reading and to analyze the implementation/integration, policies, procedures and evaluation techniques of a Bring Your Own Device (BYOD) Program in a Title I school. The research targeted the persistent problem of academic achievement gap between students from low income families and their peers from high income families in the areas of mathematics and reading which can further be exacerbated by the limited or restricted access to the use of mobile devices as learning tools. The study also examined the implementation/integration challenges of Bring Your Own Device (BYOD) programs. Of concern was how mobile learning devices in the classroom can help students from low-income families improve student achievement in mathematics and reading. Moreover, the study investigated the implementation and integration challenges schools face when using technology in schools. The research was conducted at a Title 1 middle school with a successful history of implementing a BYOD Program. The school district is recognized throughout the state, country and internationally for their Bring Your Own Technology (BYOT) Program. The data analysis from the semi-structured interviews conducted with the Principal, Technology Support Specialist and open ended survey responses from three Mathematics teachers, two Reading teachers and one English Language Arts teacher and document analysis and observation notes produced four themes that were aligned with the theoretical framework. The findings of the research explain how the school uses mobile technology as an instructional strategy in conjunction with researched based teaching practices to improve student achievement in the areas of mathematics and reading. Also, the findings of the research explain how the implementation of a well-developed Professional Development Program, along with technology training, is essential to providing support for teachers to enhance their instruction by integrating technology usage in classrooms.
    • Disruptive Innovation For Student Success At A Twenty-first Century Four-year Baccalaureate Institution

      Holloway, Kelly L.
      The problem targeted by this study is the widening imbalance between admissions and graduation rates occurring at many long-standing institutions of higher education. This study applied the theory of disruptive innovation to gain insight into the policies, procedures, beliefs, and perspectives of how a single college that was established in the twenty-first century adopted and enacted practices necessary to maintain an institution with consistent retention and competitive graduation rates. The study used an explanatory, qualitative single case study design to collect and analyze the data. The data collection methods included semi-structured, open-ended interviews and a focus group, and the review of nine relevant documents. Participants included 8 administrators, 1 faculty participant, as well as 5 faculty participants in a focus group. The research questions asked about the perceptions and influence of the mission, vision, and operating principals at the twenty-first century institution built on an innovative model. Four salient themes emerged from the data including Accessibility, Innovation, Attentiveness, and Student Success. The researcher concluded that student success measures permeated the findings of this research study. Recommendations for further study include: a longitudinal study of Innovative College, the examination of long-standing institutions that have adapted their cultures toward an accountable and innovative framework, an extension of this study to include student perceptions, and the examination of the effects of changes in leadership on the sustainability of Innovative College.
    • DKK1's Potential Role As A Biomarker In Pancreatic Adenocarcinoma

      Igbinigie, Eseosaserea Grace
      Dickkopf-1 (Dkk1)’s dysregulation has been implicated in the pathogenesis of a variety of cancers. It is part of the Dkk family of proteins that includes Dkk2, Dkk3 and Dkk4. This family of secreted proteins shares similar conserved cysteine domains and inhibits the Wnt/b-catenin pathway by causing the degradation of �?�-catenin, thereby stopping cell proliferation. Dkk1 has also been previously shown to affect the CKAP/Akt pathway to increase Akt phosphorylation and promote cell proliferation. To determine the location and pathway that Dkk1 may regulate in pancreatic cancer cells, we performed immunofluorescence assays on Suit-2 cells. The results showed that Dkk1 is mainly located in the nucleus with a small percentage of the proteins in the cytoplasm. For Dkk1’s potential receptors, CKAP4 was found to have a similar staining to Dkk1 while Lrp6 was found to be evenly spread through the nucleus and cytoplasm. Further staining with the Wnt/�?�-catenin downstream protein, �?�-catenin, showed that it was colocalized with Dkk1 in the nucleus indicating that Dkk1’s presence did not inhibit its ability to translocate into the nucleus. Further studies into the cause of Dkk1’s inability to degrade and stop �?�-catenin’s translocation that causes increased cell proliferation is needed.
    • Does Pedagogical Knowledge Support Effective Instructional Leadership In Usa Medical Education?

      Jones, Louise M.
      ABSTRACT LOUISE M. JONES DOES PEDAGOGICAL KNOWLEDGE SUPPORT EFFECTIVE INSTRUCTIONAL LEADERSHIP IN USA MEDICAL EDUCATION? Under the direction of DR. EDWARD BOUIE Ed. D The element of instructional quality in higher education is known to be a key influencer on successful student outcomes. The instructional leader is known to impact faculty instructional quality through their practices. Typically, these leaders receive little formalized preparation for their role in building capacity in instructional quality. The investigator posited that general pedagogical knowledge (GPK) would be linked to higher levels of engagement with effective leadership practices (ELP) in the leaders directly responsible for instructional quality. This cross-sectional study drew from leaders within U.S. medical education (n=189) as a subculture of higher education, using a 32 item Likert-type rating survey instrument, Educational Leadership Practices in Higher Education (ELPHE). Reliability, validity, and model-fit studies were carried out on the data set using inter-item correlation, and structural equation modeling before scoring the ELPHE subscales and full scale. The findings indicate that participants with a higher level of self-reported GPK were also more likely to have a higher level of engagement with evidence-based effective educational leadership practices. The findings from this study provide some insight into the type of knowledge and degree of mastery that best equips the leaders with direct responsibility for building capacity in faculty teaching excellence.