Search

One of the biggest challenges the clinical research industry currently faces is the accurate forecasting of patient enrollment (namely if and when a clinical trial will achieve full enrollment), as the stochastic behavior of enrollment can significantly contribute to delays in the development of new drugs, increases in duration and costs of clinical trials, and the over- or under- estimation of clinical supply. This study proposes a Machine Learning model using a Fully Convolutional Network (FCN) that is trained on a dataset of 100,000 patient enrollment data points including patient age, patient gender, patient disease, investigational product, study phase, blinded vs. unblinded, sponsor CRO selection, enrollment quarter, and enrollment country values to predict patient enrollment characteristics in clinical trials. The model was tested using a dataset consisting of 5,000 data points and yielded a high level of accuracy. This development in patient enrollment prediction will optimize portfolio demand planning and help avoid costs associated with inaccurate patient enrollment forecasting.

Inflammatory bowel disease (IBD) is a disease that is classified into two subtypes: ulcerative colitis (UC) and Crohn’s disease (CD). Symptoms can range from mild discomfort to requiring surgical intervention and affects approximately 1-in-200 adults in America alone, with global incidence rates increasing. While many treatments exist for IBD, perhaps the main reason for the lack of a cure is that there are many different pathogeneses that all lead to a very similar expression of symptoms. Over 240 IBD loci have been identified to date, yet the causative allele that drives the association has only been identified in ~60 of these 240. Even with the known IBD loci, it is estimated that the known heritability for CD and UC is minimal at 13% and 8%, respectively. Genetic risk research across many populations around the world show different rates, or even complete absence, of known risk loci identified is Caucasian populations. This wide irregularity in genetic risk across populations gives reason for additional genomic research both within and outside of previously focused populations. In this study, we approach two novel methods for the discovery of additional risk loci in IBD. One method utilized multi-omic data (e.g. whole exome sequencing, methylation, and RNA-seq) and machine learning approaches in a discordant/concordant sib-pair study. The machine learning methods used were iClusterPlus and MoCluster. The iClusterPlus analysis yielded four “top priority” genes with GBP2 as a novel discovery in IBD. MoCluster yielded 10 “top priority” genes with CSK as a novel discovery in IBD. Additionally, the MoCluster analysis identified five KEGG pathways as having strong relation with IBD: "Platelet activation"; "Viral protein interaction with cytokine and cytokine receptor"; "NF-kappa B signaling pathway"; "Ferroptosis"; and "Epithelial cell signaling in Helicobacter pylori infection". A second method utilized the largest African American population of IBD patients to date in a dense endophenotype study. Using a stratified analysis to test for endophenotype-independent risk loci yielded five genes of interest. Three of the genes, TNFSF8, HLA-DQB1, and CHRNA4, were shown to increase risk of disease and two of the genes, ADCY7 and LSAMP, were found to be protective. In this study, I have identified novel genes and pathways for further IBD research. These methods have the potential to greatly increase the known genetic causes for IBD and other diseases by expanding the classifications of disease and utilizing these distinctions. An increased number of genetic loci with known, specific disease progressions will enable physicians with enhanced targeted screening capabilities, leading to an increase in quality of life for affected patients.

This research focuses on the development of a low dose radiotherapy (LD-RT) device for treatment of acute respiratory distress syndrome (ARDS), a respiratory illness that affects millions of people a year. The use of orthovoltage (200 – 500 keV) X-ray energy provides many advantages over traditional radiotherapy delivery with linear accelerators, such as low cost and greater accessibility. In addition, X-ray tubes have been shown throughout history to provide good treatment outcomes for pneumonia, and research has shown LD-RT to be just as effective with ARDS. This proposal summarizes my efforts in determining the dosimetric properties of a LD-RT system to deliver treatment in a fast and effective manner. My research involves Monte Carlo transport simulations in MCNP to calculate the dose delivery and coverage of a whole lung treatment. In addition, different setup geometries and beam modifications will be explored to optimize the treatment delivery. The development of such a system would greatly expand the treatment options for ARDS type illness at a lower cost and greater accessibility than current radiotherapy options.

The ideal “off the shelf” tissue engineering, small-diameter (< 6 mm inner diameter (ID)) vascular graft hinges on designing a template that facilitates transmural ingrowth of capillaries to regenerate an endothelized neointimal surface. Previous traditionally electrospun (TES) approaches to create bioresorbable vascular grafts lack the pore sizes required to facilitate transmural capillary ingrowth required for successful in situ neovascular regeneration. Therefore, the ability to create scaffolds with program-specific architectures independent of fiber diameter via the relatively recent sub-technique of near-field electrospinning (NFES) represents a promising solution to create tissue engineering vascular grafts. These programmed large pore sizes are anticipated to promote in situ regeneration and improve the outcomes as well as the quality of life of patients with arterial disease. In this dissertation, we manufactured via NFES as well as characterized biodegradable polydioxanone (PDO) small-diameter vascular grafts. Chapter 1 introduces the need for off-the-shelf, small-diameter vascular grafts to facilitate in situ regeneration, the process and pore size limitations of TES vascular grafts, and the promising use of NFES to develop precisely tailored PDO vascular grafts. Chapter 2 describes the process of NFES and details the current progress in NFES of biomedical polymers as well as the major limitations that exist in the field. Chapters 3, 4, and 5 contain primary research exploring the creation of an NFES vascular graft scaffold and characterizing the mechanical as well as biological response of these scaffolds. Specifically, in Chapter 3 we demonstrate a NFES apparatus designed around a commercial 3D printer to write PDO microfibers. The processing parameters of air gap, polymer concentration, translational velocity, needle gauge, and applied voltage were characterized for their effects on PDO fiber diameter. The processing parameters of polymer concentration and translational fiber deposition velocity were further characterized for their effects on fiber crystallinity and individual fiber uniformity. The precision of fiber stacking via a 3D printer was qualitatively evaluated to inform the creation of 3D scaffolds to guide the alignment of human gingival fibroblasts. It was found that fiber diameters correlate positively with polymer concentration, applied voltage, and needle gauge and inversely correlate with translational velocity and air gap distance. Individual fiber diameter variability decreases, and crystallinity increases with increasing translational fiber deposition velocity. These data resulted in the creation of tailored PDO 3D scaffolds which guided the alignment of primary human fibroblast cells. Together, these results suggest that NFES of PDO can be scaled to create precise geometries with tailored fiber diameters for vascular graft scaffolds. In Chapter 4, we demonstrated a NFES device to semi-stably write PDO microfibers. The polymer spinneret was programmed to translate in a stacking grid pattern, which resulted in a scaffold with highly aligned grid fibers that were intercalated with low density, random fibers. As a consequence of this random switching process, increasing the grid dimensions resulted in both a lower density of fibers in the center of each grid in the scaffold as well as a lower density of “rebar-like” stacked fibers per unit area. These hybrid architecture scaffolds resulted in tailorable as well as greater surface pore sizes as given by scanning electron micrographs and effective object permeability as indicated by fluorescent microsphere filtration compared to TES scaffolds of the same fiber diameter. Furthermore, these programmable scaffolds resulted in tailorability in the characterized mechanical properties ultimate tensile strength, percent elongation, yield stress, yield elongation, and Young’s modulus independent of fiber diameter compared to the static TES scaffold characterization. Lastly, the innate immune response of neutrophil extracellular traps (NETs) was further attenuated on NFES scaffolds compared to TES scaffolds. These results suggest that this novel NFES scaffold architecture of PDO can be highly tailored as a function of programming for small diameter vascular graft scaffolds. In Chapter 5, we created two types of NFES PDO architectures, as small-diameter vascular graft scaffolds. The first architecture type consisted of a 200 x 200 µm and 500 x 500 µm grid geometry with random fiber infill produced from one set of processing parameters, while the second architecture consisted of aligned fibers written in a 45°/45° and 20°/70° offset from the long axis, both on a 4 mm diameter cylindrical mandrel. These vascular graft scaffolds were characterized for their effective object transit pore size, mechanical properties, and platelet-material interactions compared to TES scaffolds and Gore-Tex® vascular grafts. It was found that effective pore size, given by 9.9 and 97 µm microsphere filtration through the scaffold wall for NFES grafts, was significantly more permeable compared to TES grafts and Gore-Tex® vascular grafts. Furthermore, the characterized mechanical properties of ultimate tensile strength, percent elongation, suture retention, burst pressure, and Young’s modulus were all tailorable for NFES grafts, independent of fiber diameter, compared to TES graft characterization. Lastly, platelet adhesion was attenuated on large pore size NFES grafts compared to the TES grafts which approximated the low level of platelet adhesion measured on Gore-Tex® grafts, with all grafts showing minimal platelet activation given by P-selectin surface expression. Together, these results suggest a highly tailorable process for the creation of the next generation of small-diameter vascular grafts. Lastly, Chapter 6 expounds future considerations for continuing research in NFES technology, NFES for general tissue engineering, and NFES for vascular tissue engineering as well as gives final conclusions. Together, the finding of this dissertation indicated that NFES vascular grafts result in seamless, small diameter tubular scaffolds with programmable pore sizes on the magnitude anticipated to facilitate transmural endothelialization as well as programmable mechanical properties that approximate native values. Thus, this work represents the next step in developing bioinstructive designed scaffolds to facilitate in situ vascular regeneration to improve the outcomes as well as the quality of life of patients with arterial vascular disease.

The prevalence of prediabetes and diabetes in the United States and around the world has increased faster than expected in the last 30 years. The economic burden this costs a nation can be astronomic both in terms of expense and loss in productivity. One-third of U.S. adults, 86 million people, have prediabetes. Effective management is needed that can reach these 86 million, and others at high risk, to reduce their progression to diagnosed Type 2 diabetes. After the literature review, there was not enough literature to support how these led to the progression to diabetes. The abundant literature is centered on how to prevent complications and improve the quality of life of those living with type 2 diabetes. This paper will focus on the longitudinal association between these social determinants and how they may predispose to the progression to Type 2 diabetes.

In 1912, after having lived on the island since the Civil War, the mixed-race community of Malaga Island, located just off of the coast of Phippsburg, Maine, were forcibly removed from their homes. Some of those who were removed were sent to the School for the Feeble-Minded in Pownal, Maine. The forced removal of the residents and the sterilization of those who were institutionalized were inextricably grounded to the notions of race, gender, and eugenics. Although Gov. Frederick Plaisted at the time deemed the dismantling of the island as critical in raising property values and the tourism industry, it’s clear that these were simply excuses to rid the island of its residents. Maine was ashamed of Malaga’s mixed-race community and used the term, “malagite” as a racial slur (Maine State Museum, 2021). Examining scholarly articles on eugenics, race, and gender I will explore the constructions behind the eradication and the relocation of the Malaga Islanders. As an intern for the Maine Womens Magazine, I will attempt to write an article that will showcase these findings. The tensions that may arise as Maine residents respond to this article, will also be explored as indicative of the continual eradication and dismissal of Malaga Island residents and Maines’ racist past.

In the quantum logic of the DNA molecule, electrons are held and conducted coherently as spinless Cooper pairs and are shielded from electromagnetic energy by a Faraday cage effect of the double lipid bilayer of the nuclear membrane. The magnetic vector potential generated by cellular depolarization can synchronize logical activity in portions of the DNA molecule by affecting spin directions of appropriately oriented spinless electrons via the Aharonov-Bohm effect, but is not blocked by that Faraday cage effect. Within the logically and thermodynamically reversible chiral enantiomeric symmetry of the deoxyribose moieties the decoherent transition of Cooper pair to Dirac pair in a p-orbital of the C2-C3 covalent bond effects chiral selection between the C2-endo and C3-endo conformations. Such a spin-1/2 chiral collective movement of particles can be considered as a quasiparticle excitation that is its own antiparticle (C2-endo vs. C3-endo), meeting the definition of a Majorana fermion.

The DNA molecule can be modeled as a quantum logic processor, and this model has been supported by pilot research that experimentally demonstrated non-local communication between cells in separated cell cultures. This modeling and pilot research have important implications for information sciences, providing a potential architecture for quantum computing that operates at room temperature and is scalable to millions of qubits, and including the potential for an entanglement communication system based upon the quantum DNA architecture. Such a system could be used to provide non-local quantum key distribution that could not be blocked by any shielding or water depth, would be simultaneous over any distance, and could not be electromagnetically interfered with or eavesdropped upon. The quantum DNA model also has implications for artificial neural networks and can provide architecture for a system of quantum random number generation.

Abstract Background and Significance: Managing pain is a challenge for many organizations, including the U.S. military. The added complication of managing sleep disruption compounds the issue. Examining pain and sleep disruption in the context of work performance within the military is critical to understanding the impact of these conditions on our nation’s fighting forces. Purpose: Examine the associations of chronic pain and sleep disruption in the context of work performance among active duty military service members. Research Aims: 1) Describe sociodemographic characteristics, military service characteristics, clinical characteristics, and work performance of active duty service members, 2) Examine relationships between these characteristics, and work performance, and 3) Determine what characteristics increase task performance and contextual performance, and decrease counterproductive work behavior. Design: A cross-sectional observational study that examines associations between patients with chronic pain and sleep disruption, in the context of work performance. Methods: Subjects were recruited and consented from the empaneled pain management clinic. Participants completed a single visit in which self-reported demographics, pain status, sleep status, and work performance were captured. Findings: 145 participants completed the study. Females accounted for 27.6% (n = 40) of participants, exceeding the 14.4% overall military average. Special Forces participants had higher levels of opioid use and depression compared to the rest, but demonstrated better work performance. Age, depression, and sleep were consistently strong predictors of work performance. Patients performed better with age, while those with depression and sleep disruption performed poorly. Implications: A closer look at the Special Forces population in the context of pain, sleep, and work performance could provide a better understanding of how these patients cope with pain and sleep disruption. Research focused on the differences in work performance among age groups may provide a better understanding of coping strategies. Focused depression research can lead to a greater understanding of how mental health impacts pain, sleep, and work. Overall, this study revealed some interesting relationships that may lead to interventions within this population. The findings of this study opens the door to explore multiple approaches that could lead to treatments and preventions for military members suffering from chronic pain.

Context: Blood flow restriction (BFR) has previously been used in a rehabilitation setting to decrease the external load applied to post-surgical patients and those with chronic debilitating conditions. BFR in a performance context has been explored minimally, particularly as it applies to baseball pitching over time. Objective: The purpose of this study was to retrospectively investigate the trend in throwing velocity of fastballs with BFR implementation over one competitive season. Design: This study was a retrospective longitudinal design on pitching velocity of fastballs throughout a competitive season. Setting: The data collection and analysis was completed at a mid-Atlantic university. Patients or other participants: Four Division I starting baseball pitchers. Inclusion criteria were a starting baseball pitcher during the 2018-2019 season, on the official baseball roster for that season and had a sport physical on file. Exclusion criteria were any individual not meeting the inclusion criteria. Intervention: Data analysis was performed using data recorded by Trackman during home games throughout the 2018-2019 competitive baseball season. The fastball pitch velocity data was deidentified and participants were assigned a subject number. BFR was applied by the full-time baseball athletic trainer to all 4 pitchers following pitching bouts during all 60 games during the 2018-2019 baseball competitive season. The BFR occlusion protocol varied based on the limb size/arm girth of the individual, with a goal of reaching approximately 50 percent of the limb occlusive pressure. The cuff was then be inflated for five minutes with no concomitant exercise and then deflated for one minute. This process of inflation followed by deflation was repeated three times, for a total treatment duration of 18 minutes, in which, the cuff was inflated for 15 minutes. Main Outcome Measures: Pitching velocity of fastballs, number of fastballs thrown and number of innings pitched were all recorded and analyzed for each subject. Results: The highest fastball pitch velocity average was 96.0 ± 0.9 mph, which occurred during game 11 in inning 1 and inning 3 (96.0 ± 0.7 mph), pitched by subject 1. The lowest fastball pitch velocity average was 86.4 ± 1.2 mph, which occurred during game 4, inning 2 pitched by subject 2. In all games except for games 2, 6, 7, 10 and 14, the fastball pitching velocity over the game remained consistent with a slight decrease. In all games except for games 2, 6 and 15, the fastball pitching velocity averages by inning over the game remained relatively consistent with a decrease. Games 7 and 10 depicted the most consistent fastball pitches thrown over the course of a game, with no detectable increase or decrease from the line of best fit. Subject 1 pitched in four games and had the highest overall pitch velocity average throughout the 4 games with 92.45 ± 0.95 mph and was also the most consistent fastball pitcher on average. Subject 3 exhibited the second highest overall pitch velocity over a span of 5 games, with an average pitch velocity of 92.06 ± 1.2 mph. Next was subject 4, with an average pitch velocity of 88.86 ± 1.0 mph. Causality could not be determined whether the consistency in fastball velocity was related to the use of a BFR recovery treatment due to deviations from the BFR protocol outlined in the intervention. Conclusions: The number of fastballs thrown decreased throughout a single game progression, but the average number of fastballs thrown throughout the season remained consistent from game to game. The average pitching velocity of fastballs thrown remained consistent throughout individual games. Furthermore, average fastball pitching velocity remained consistent with a slight increase in throwing velocity among individual subjects.

Undiagnosed individuals with signs and symptoms of obstructive sleep apnea (OSA) are at higher risk of severe health complications than healthy individuals, and they pose a huge economic burden on society as well. Some scholars reported the lack of public knowledge of OSA as a major contributor to this phenomenon. However, there is a lack of information about assessing public knowledge and health belief of OSA and its nature across the risk level of having OSA. Two self-reporting questioners were used in this mixed method study, a PI developed survey, the Obstructive Sleep Apnea Knowledge and Belief (OSA-KAB) © was used assess adults’ knowledge and health beliefs surrounding OSA, and the STOP-Bang questionnaire that was used to categorized individuals based on their risk level of developing OSA. The quantitative data result indicated a sever lack of knowledge of OSA across all seven subscales and was confirmed by the qualitive data. Additionally, while there was a significant, positive relationship between the knowledge and health belief, neither knowledge nor health belief scores were influenced by the risk level of OSA. The results of this support a definite need to increase public knowledge about OSA, specifically risk factors and complications due to their impact on health belief. In addition, business owners need to identify the economic value of addressing their employees’ risk of OSA, as the cost of diagnosing and treating OSA was identified as a major barrier to usage.

Familial hypercholesterolemia (FH) is a common genetic disease and has been studied with the aim of finding a curative measure for decades. FH is caused by mutations in the low-density lipoprotein receptor (LDLR) resulting in defects in LDL-cholesterol (LDL-C)-receptor mediated endocytosis and development of premature cardiovascular disease (CVD). Here I describe the use of a corrected and non-corrected LDLR FH cell model to investigate receptor-mediated endocytosis and statin effects. For these studies, we reprogrammed FH fibroblast cells to induced pluripotent stem cells (iPSC) and confirmed their pluripotency and ability to differentiate to hepatocyte-like cells (HLC). A clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) design was created and used to successfully correct a 3 base pair deletion in the class II LDLR FH iPSC. Western blot and immunocytochemistry analysis showed maturation and proper localization of the LDLR after correction in iPSC and HLC. We used internalization analyses to detect LDL-C receptor mediated endocytosis in non-corrected and corrected HLC. Real time PCR for ER stress markers showed that statin-induced accumulation of an immature LDLR in non-corrected FH cells did not activate the unfolded protein response. Together, this work demonstrates the capacity to utilize the resources of CRISPR, iPSC, and HLC to study misfolded class II LDLR providing a physiologically relevant in vitro model for investigating the differential effects of mutant versus corrected LDLR-mediated metabolism and current treatments like statins.

The structures of the spinal cord and vertebral column are designed to provide flexibility, while still providing ample protection for the spinal cord deep within. While it does offer remarkable protection against most routine trauma, the spinal cord is still vulnerable to high-force etiologies of trauma and may become damaged as a result. These events are referred to as primary injury. Following the initial injury, the body’s own physiological responses cause a cascade of deleterious effects, known as secondary injury. Secondary injury is a major therapeutic target in mitigating the effects of spinal cord injury (SCI), and much research is currently being done to develop more effective treatment options.

Radioembolization of liver tumors using yttrium-90 (Y-90) labeled microspheres is a proven method of treatment. The primary decay mode of Y-90 is beta-minus decay. The electrons produced during decay deliver a cytotoxic radiation dose in a localized volume. The radiation dose is sufficient to kill tumor cells yet spare the normal liver. The Y-90 microspheres are delivered via a catheter that has been placed in an artery that supplies blood to the tumor. Microspheres are injected and the blood distributes the microspheres. The current practice involves two separate procedures. The first procedure is an assessment of the vasculature of the liver and tumor. This assessment is performed with radiolabeled macro-aggregated albumin particles (Tc-99m MAA). The Tc-99m MAA is injected via a catheter placed in the same location that will be used for the delivery of Y-90 microspheres. The distribution of the Tc-99m MAA particles is imaged using a gamma camera. This data is used to predict the microsphere distribution and subsequent dose to the tumor, normal liver, lungs and other organs. The data may be used to adjust the Y-90 activity and to perform crude pre-treatment dosimetry. Accurate pre-treatment dosimetry is largely absent. This study was performed to create a positron-emitting surrogate microsphere that was close in size and shape to the Y-90 microspheres. Positron emission tomography (PET) is superior to SPECT for activity quantification. Fluorine-18 (F-18) was chosen for initial experiments. There were two aims of the research: Aim 1: To develop surrogate PET microsphere. This included radiolabeling, stability testing in vitro and stability testing in an animal. Aim 2: To develop a computational technique to perform dosimetry with the surrogate microsphere. PET images of the surrogate radionuclide were imported into a Monte Carlo program. PET images were used to simulate the distribution of the Y-90 microspheres. The output of the simulation was a tomographic map of absorbed dose from the Y-90. Validation of the image-based dosimetry was performed by measuring known amounts of F-18 and Y-90 in a phantom. The measured dose within the phantom was compared to the dose predicted with the Monte Carlo simulation.

The Bragg peak geometry of the depth dose distributions for hadrons allows for precise and effective dose delivery to tumors while sparing neighboring healthy tissue. Further, compared against other forms of radiotherapeutic treatments, such as electron beam therapy (EBT) or photons (x and \(\gamma\)-rays), hadrons create denser ionization events along the particle track, which induces irreparable damage to DNA, and thus are more effective at inactivating cancerous cells. The measurement of radiation's ability to inactivate cellular reproduction is the relative biological effectiveness (RBE). A quality related to the RBE that is a measurable physical property is the linear energy transfer (LET), a measurement of the deposited energy over the particle track. The purpose of this work was to develop a method of measuring LET of radiotherapeutic beams using an air-filled ionization chamber (IC) and liquid-filled ionization chamber (LIC). Through analysis of the IC and LIC signals, we determined a correlation between the recombination index (IC/LIC) and dose-averaged LET.

Protein Kinase D1 (PKD1) is a serine threonine kinase which is downregulated in Prostate, Breast and Colon Cancer. It functions as a tumor suppressor in different cancer cells. Downregulation of PKD1 is known to be associated with aggressiveness of the cancer. PKD1 is known to regulate many key oncogenic signaling pathways such as E-cadherin, β-catenin and Androgen Receptor signaling pathways. Aberrant expression of these oncogenic pathways leads to transformation of cells from normal to malignant phenotype, thereby leading to increased proliferation, growth and metastasis to distant organs of these cancer cells. Literature evidence also points to the fact that E-cadherin β-catenin and PKD1 play a role in regulation of epithelial mesenchymal transition (EMT). To fully understand how PKD1 regulates β-catenin signaling, we investigated the effect of PKD1 overexpression on β-catenin signaling in colon cancer cells. We observed that PKD1 overexpression is responsible for inhibition of cell proliferation and colony formation ability of different colon cancer cell lines. Moreover, nuclear PKD1 overexpression leads to inhibition of β-catenin transcription activity in colon cancer cells. Further evaluation in in vivo mouse model showed that PKD1 is responsible for inhibition of colon cancer tumor growth in xenograft mouse model. This paved way for us to look for the effect of PKD1 on other downstream targets of β-catenin pathway which regulate EMT process in cancer cells such as Metastasis associated Protein 1. Metastasis associated Protein 1 (MTA1) is a nucleosome remodeling and histone deacetylase protein (NuRD) which is overexpressed in all the cancers. MTA1 is an initiator of epithelial and mesenchymal transition and is responsible for cancer cells metastasizing to different organs of the body. Expression of MTA1 directly correlates with the aggressiveness of the cancer. MTA1 is known to regulate β-catenin and Androgen Receptor signaling pathways leading to cancer cells acquiring metastatic capabilities. Therefore, in our study we evaluated the inverse correlation between MTA1 and PKD1 in different cancer cells. To investigate the cellular effect of PKD1 in prostate and colon cancer, stable PKD1 overexpressing prostate (C4-2) and colon cancer cells (SW480) were utilized. PKD1 overexpression inhibited MTA1 expression in prostate and colon cancer cells. PKD1 interacts, phosphorylate, translocate and degrades MTA1. Kinase domain and N terminal domain of PKD1 play a significant role in MTA1 interaction and phosphorylation. Phosphorylation of MTA1 leads to nuclear export via golgi and trans-golgi network to lysosome. Bryostatin-1 is a macrocyclic lactone which modulates PKD1 activity. Bryostatin-1 was used to activate PKD1 expression in C4-2 cells and MTA1 translocation was then tracked. This translocation of MTA1 to lysosome is a ubiquitin dependent phenomenon leading protein degradation. PKD1 overexpression leads to inhibition of tumor growth and bone metastasis leading to inhibition of osteoblast to osteoclast formation as determined by RANK expression. PTEN Knockout and TRAMP mouse model also show inverse correlation between PKD1 and MTA1 expression in prostate tissues at different weeks. Human tissue microarray of prostate, colon and breast cancer (MTA1 is overexpressed and PKD1 is downregulated in breast cancer, therefore, we tested our hypothesis in breast cancer as well) showed inverse correlation between PKD1 and MTA1 in different grade tumor tissue signifying clinical relevance of this correlation. For proof of concept of our hypothesis we used ormeloxifene because Bryostatin-1 has mild toxicity issue. Ormeloxifene is a novel modulator of PKD1 activity and it targets rapidly dividing cells Further, we investigated the effect of ormeloxifene on activation of PKD1 leading to inhibition of cancer metastasis. We observed specific activation of PKD1 expression of ormeloxifene which inhibited MTA1 expression leading to inhibition of tumor growth in xenograft mouse. We further evaluated the efficacy of ormeloxifene to inhibit metastatic prostate cancer cells (PC3 and DU145). Ormeloxifene showed excellent anti-cancer efficacy against prostate cancer as it inhibited cell proliferation, invasion and migration of metastatic prostate cancer cells. Moreoever, ormeloxifene induced cell cycle arrest at G0/G1 phase by regulating key cell cycle regulatory proteins. It also inhibited metastasis of prostate cancer leading to inhibition of key metastatic markers involved to epithelial mesenchymal transition. Ormeloxifene also showed excellent in vivo efficacy against metastatic prostate cancer cells. Therefore, ormeloxifene could be a potential therapeutic modality for metastatic cancers as it targets EMT signaling. To conclude, we for the very first time have elucidated a novel regulatory mechanism of PKD1 mediated regulation of MTA1 that plays an important role in cancer progression and metastasis. For cancer cells to metastasize PKD1 expression is suppressed with subsequent increased expression of MTA1. We elucidated that repression of MTA1 with subsequent activation of MTA1 leads to attenuation of cancer metastasis. Moreover, therapeutic modality that targets this novel regulatory pathway leading to activation of PKD1 and inhibition of MTA1 is an ideal candidate for treatment of advanced stage metastatic cancers.

The plasma membrane protein Efr3b, the Tetratricopeptide domain repeat 7 (Ttc7), and the kinase phosphatidylinositol 4-kinase III alpha (PI4KIIIa) are found to form a complex a the plasma membrane. This complex is thought to play a role regulating the PI4P pools at the plasma membrane, which is imperative to membrane identity and vesicle transport.

Originally, God never put sickness on people. After the fall, the early history of man as recorded in the Bible further demonstrates Him refraining to use sickness. However, once the law was instituted God began to use sickness as a punishment. In the Old Testament, sickness was always defined as a curse and never a blessing. It was used by God to curse people for disobeying the law. In the New Testament, God was seen healing the multitudes instead of cursing them. Through the atonement of Jesus, believers have been redeemed from the curse of the law (Galatians 3:13). Healing has been provided with the forgiveness of sins. There are numerous reasons why people still are not healed but it is not God’s Will for them to remain ill.

Polymers have revolutionized the field of tissue engineering due to the countless possibilities of scaffolds that can be constructed from such a material. Through conjugation of other functional groups onto a polymer backbone, the mechanical properties of polymers can be controlled and has the ability to span a wide range of strengths, allowing for application in multiple tissue environments. One such polymer, hyaluronic acid (HA), is a natural polysaccharide found most abundantly in the extracellular matrix (ECM) of connective tissues with structural, lubricating, and wound healing functions in the human body; for this reason, HA has great potential for utilization in tissue engineering application. To strengthen the mechanical properties of HA, we conjugated 2-aminoethyl methacrylate (AEMA) to the carboxyl group of HA polymer in various degrees of substitution. In addition, different molecular weights of HA were used during synthesis to analyze molecular weight as a factor for tunable mechanical properties. The final product HA-AEMA, with use of photoinitiator Irgacure 2959, forms a hydrogel via UV polymerization following dissolution of macromer into photoinitiator solution. Both the rheological and mechanical properties of various macromer/photoinitiator solutions and hydrogels were measured, respectively, in regards to viscosity, solubility, shear modulus, and compression strength. To test HA-AEMA potential for use of tissue engineering applications involving stem cells, human dental pulp stem cells (DPSCs) were exposed to both HA-AEMA macromer and hydrogels for cytotoxicity and encapsulation studies by MTT using mitochondrial based assays and fluorescent viable cell staining respectively. Results of our experiments indicate successful synthesis of HA-AEMA macromer of various molecular weights of HA as well as degree of substitution of AEMA. As the molecular weight of HA and/or the degree of substitution of AEMA increased, the viscosity, swelling ratio, shear storage modulus, and compressive strength increased; specifically for storage modulus, more significant increases were observed as MW of HA was increased from 16 kDa to 66 kDa compared to increases from 66 kDa to 270 kDa. As the hydrogels swelled over time in phosphate buffer solution (PBS), the compressive strength of hydrogels diminished by a factor of 3 after 24 hours of swelling time and a factor of 2 from 1 day to 3 days swelling time, most likely due to hydrolysis of the ester bond along the HA chain. Results of the cytotoxicity studies indicate DPSCs survive exposure to HA-AEMA macromer as well as encapsulation inside hydrogels formed following the standard UV polymerization protocol. Based on these results, we have concluded that HA-based hydrogels have the potential to be used in various tissue engineering applications due to its tunable properties and biocompatibility to DPSCs

In the inner and outer hair cells (OHCs) of the inner ear, an unconventional myosin 15a localizes at the tips of mechanosensory stereocilia and plays an important role in forming and maintaining their normal structure. A missense mutation makes the motor domain of myosin 15a dysfunctional and is responsible for the congenital deafness DFNB3 in humans and deafness and vestibular defects in Shaker-2 (Sh2) mouse model. All hair cells of homozygous Shaker-2 mice (Myo15sh2/sh2) have abnormally short stereocilia, but, only stereocilia of Myo15sh2/sh2OHCs start to degenerate after the first few days of postnatal development and lose filamentous tip links between stereocilia that are crucial for mechanotransduction. The exact mechanisms of this degeneration are unknown even though they may underlie DFNB3 deafness in humans. We hypothesize that structural abnormalities in Myo15sh2/sh2 OHCs may alter the mechanical forces applied to the mechano-electrical transduction (MET) channels resulting in abnormal ionic homeostasis, which may lead to eventual degeneration of Myo15sh2/sh2 OHCs. Therefore, we investigated the ionic conductances and integrity of mechanotransduction apparatus in Myo15sh2/sh2 OHCs. Surprisingly, we found that myosin 15a-deficiency is associated not only with structural abnormalities of OHC stereocilia but also with alterations of voltage-gated ion conductances.

Context: A pre-participation examination (PPE) has become standard practice among the athletic community. This examination commonly includes a multifaceted baseline concussion assessment and an injury history survey. Recent evidence suggests that neuropsychological testing can aid in predicting individuals at an increased risk of lower extremity injury. However, no known previous study has investigated the relationship between neuropsychological function and potential risk of sustaining a concussion. Objective: This study sought to identify a relationship between components of a standard PPE and an elevated risk of concussion. Design: All data was extracted from the institution concussion database. Setting: A large university in Southeast Georgia. Participants: One hundred and sixty-six participants were recruited for this study, of these participants eighty-two were in the concussed experimental group and eighty-two were in the matched healthy control group, with two excluded for invalid or incomplete ImPACT data. Main Outcome Measurements: Statistics included a descriptive analyses of gender, sport, and concussion history, a frequency analysis of the four ImPACT composites, total BESS score, total SAC score, gender, sport, and injury history, a series of one-way ANOVAs, a ROC analysis, and a discriminant function analysis. Results: The frequency analysis determined that there was some missing data, the descriptive analysis determined the following group means, verbal memory composite: 85.7 +/- 11, visual memory composite: 73.1 +/- 16.3, reaction time composite: 0.577 +/- 0.080, processing speed composite: 39.8 +/- 7.7, BESS: 13.1 +/- 6.1, SAC: 27+/- 2. The series of nine one-way ANOVAs showed no significant group differences. The ROC analysis determined the following cut off values for each PPE component, verbal memory composite: 83.5, visual memory composite: 81.5, reaction time composite: 0.63, processing speed composite: 33.05, BESS: 13, and SAC: 26. The discriminant function analysis revealed no significant predictors. Discussion: This study began to show that the basic components of the PPE may not be clinical predictors of concussions. This is clinically significant because it rules out the baseline assessment approach as something that could help identify individuals at an increased risk.

Patient safety is one of the most important health care challenges. It is a big concern since 1 in every 10 patients around the world is affected by healthcare errors. The focus of this study is given to preventable adverse events that caused by the errors or system flaw that could have been avoided. In this study, simulation models are developed using Arena to evaluate the impact of GS1 data standards on patient risk in healthcare supply chain. The focus was given to the provider hospital supply chain operations where inventory discrepancy and performance deficiencies in recall, return, and outdate management can directly affect patient safety. Simulation models are developed for various systems and scenarios to compare different performance measures and analyze the impact of GS1. The results indicates that as the validation points are closer to the point of use, the number of recalled or outdated products administered to a patient are still reduced significantly so checking at the bedside or PAR is critical. But validation only at these points may cause some problems such as stock outs; therefore, validating in other locations is also needed.

.

The increasing prevalence of antibiotic-resistant bacteria, including Mycobacterium tuberculosis, Streptococcus pneumoniae, Staphylococcus aureus, and Enterococcus faecalis, pushes us to discover new antibacterial agents to maintain adequate patient coverage. This body of work highlights the use of medicinal chemistry methodologies that encompass cross-disciplinary fields of study. Chapter 1 gives an introduction to the antibacterial drug targets, resistance, and how scientists are working to overcome obstacles encountered with drug-resistant bacteria. It also details modern medicinal chemistry applications in antimicrobial drug discovery. Chapter 2 details the use of a structure-guided library approach to drug design, in which large virtual libraries against the target are generated and filtered, based on pharmacophoric and structural constraints, to produce smaller and more structurally complex libraries prioritized for synthesis. In this work, bi-aryl sulfonamide libraries using contemporary medicinal chemistry techniques were synthesized as potential inhibitors of Mycobacterium tuberculosis cell wall biosynthesis via the rhamnose pathway. Chapter 3 describes the discovery of novel inhibitors of the PlsX/PlsY pathway to phosphatidic acid, a key intermediate in the biosynthesis of phospholipids in Gram-positive bacteria. Substrate mimics, incorporating various bioisosteric replacement head groups, were discovered demonstrating good enzyme inhibition and good antimicrobial activity against clinically relevant bacteria. Finally, Chapter 4 provides an overall discussion of the work detailed in this dissertation and future directions that will continue the advancement of these projects.

Utilizing recent advances in computer technology, Our Biomechanics Laboratory have made an effort to integrate computer animation and engineering analysis software into biomedical research, specifically towards simulation and animation of in vitro experimentation of the human cervical spine in the virtual world. The objectives of this study were to develop a virtual model of the human cervical spine for physics-based simulation and to apply the virtual model to studies of different surgical procedures and instrumentation. A process for creating an accurate virtual model of the human cervical spine was developed. The model consisted of seven vertebrae (C2-T1) connected with soft tissue components: intervertebral joint, facet joints, and ligaments. The soft tissue components were assigned nonlinear viscoelastic properties. The evaluation of the model included the percent contribution of rotation relative to global rotation, coupling behaviors, helical axes of motion pattern, global rotational stiffness curves, and animations of the disc and facet forces. This model was used to evaluate different mounting configurations for axial rotation testing and to identify a set of end constraint conditions that produced physiologic responses during axial rotational loading. This model was also used to simulate the biomechanical responses of single-level cervical fusion. The single-level fusion was found to produce increased motion compensation at the adjacent segments during flexion and extension. Greater increases in the disc forces were found in the spinal level superior to the fusion during flexion and inferior to the fusion during flexion extension. This model was also used to study of the biomechanical effects of different design features for cervical disc arthroplasty. A constrained spherical joint placed at the disc level significantly increased facet loads during extension. Lowering the rotational axis of the spherical joint into the subjacent body also caused a marginal increase in facet loading during flexion, extension, and lateral bending. Un-constraining the spherical joint to a plane at the disc level minimized facet load build up. The virtual model bridges the gap between the cadaveric-based in vitro tests and clinicalbased experimental studies to further the research and educational knowledge of cervical spine biomechanics.

Bordetella bronchiseptica colonizes the upper respiratory tract of a wide variety of mammals and is involved in diseases such as kennel cough in dogs, atrophic rhinitis in pigs and upper respiratory tract infections of laboratory animals. Studies were focused on developing a heterologous antigen expression system in B. bronchiseptica and to evaluate the potential of this organism as a candidate mucosal vaccine vector. Since the role of Bordetella bronchiseptica and Pasteurella multocida toxin (PMT) in the disease atrophic rhinitis of pigs was well documented, this research was focused in the possibility of developing a refined vaccine to control this disease. A promoter region related to heat shock protein genes was identified using a green fluorescent protein reporter system. This promoter drove high- level expression of GFP compared to a tac promoter or B. bronchiseptica fim N gene promoter. A nontoxic protective P. multocida toxin fragment and GFP were expressed in B.bronchiseptica in a broad-host-range plasmid vector PBBR1MCS2 under the control of the promoter region identified. Colonization kinetics, plasmid stability, and immune responses generated following intranasal inoculation of recombinant B. bronchiseptica were evaluated. While wild type and recombinant B. bronchiseptica colonized the mouse respiratory tract effectively, the plasmid was completely lost from the organism after 72 hours post- inoculation. After a single intranasal inoculation, B. bronchiseptica specific IgM, IgA and IgG responses were detected in serum and respiratory lavage. However, PMT-specific antibodies were not detected. Four intranasal inoculations with B. bronchiseptica expressing green fluorescent protein (GFP) induced a GFP-specific systemic and mucosal immune response, while similar inoculations with B. bronchiseptica expressing PMT fragment did not induce a PMT-specific immune response. This study also evaluated the immune response to a chimeric protein generated by combining a gene fragment encoding neutralizing epitopes of Mannheimia haemolytica leukotoxin and a fimbrial protein gene (fim N) from B. bronchiseptica. Immunization of mice with the recombinant chimeric protein elicited a significantly stronger anti-leukotoxin antibody response than comparable immunizations with fusion proteins lacking FIM N. The chimeric protein exhibited more stability. Leukotoxin is an important virulence factor in shipping fever pneumonia in feedlot cattle and is a critical protective antigen. This chimeric protein may be a candidate for inclusion in new generation vaccines against shipping fever pneumonia. The results of these studies strongly support the potential for developing B. bronchiseptica as a candidate mucosal vaccine vector and FIM N as a carrier protein for heterologous antigens.

Successful transition through meiosis is required for production of chromosomally-balanced gametes. When chromosome segregation goes awry during meiosis, aneuploidy can occur. Unfortunately, the mechanisms behind this nondisjunction are not well understood. Therefore, this dissertation has focused on learning more about the causative factors associated with aneuploidy during spermatogenesis. Are there factors that are always associated with leading to production of aneuploid sperm? One of the main goals of this dissertation is to find mouse models to study what factors may be involved in chromosome malsegregation; such as pairing, recombination, and transition through the division phases of meiosis. The first part of the dissertation will be an introduction into what is known about gamete aneuploidy. This section will review what is known about how meiotic error may arise in both humans and the mouse. The introduction will discuss links between factors that are thought to be associated with aneuploidy, and this dissertation will extend this information into new directions in analysis of predisposing factors of gamete aneuploidy. Part II focuses on a novel mouse model for gamete aneuploidy. PL/J males were found to be an important mouse model for both gamete aneuploidy and abnormal sperm-head morphology. In addition, it was found that PL/J males exhibit both genetic and phenotypic complexity in regard to the traits of aneuploidy and abnormal sperm-head morphology. Parts III-VII discuss other useful mouse models for study of gamete aneuploidy. Robertsonian heterozygous (Rb/+) translocation mice and Mlh1 -/- mice were both used to examine what happens when meiosis goes awry. For example, both Rb/+ and Mlh1 -/- mice were found to have a checkpoint that most likely detects unaligned or abnormal chromosome configurations. High percentages of MI spermatocytes in these mice were found to be apoptotic. In Part V, Brca2 -/- mice were rescued with the human BRCA2 transgene. These mice survive, but are sterile. Analysis was performed to determine the point of arrest in these mice and if they have features of a normal progression through meiosis. The last two chapters focus on different approaches for the study of aneuploidy. Part VI examines whether the topoisomerase-II inhibitor, etoposide, can induce meiotic nondisjunction. It was shown by sperm FISH that etoposide does induce meiotic nondisjunction, with the highest frequency of nondisjunction occurring at MII. The next part of this section discusses use of a novel screen for detection of new meiotic mutations. A sperm FISH screen was used in this study to detect dominant mutations. This study showed that although screening by sperm FISH is feasible, it is not a practical screen when large numbers of gametes need to be scored. The last section, Part VIII, is a summary of what we have learned and what directions should be taken to increase our understanding of how meiotic error arises leading to nondisjunction. This section will compare and contrast what we have learned from each mouse model and what factors may contribute to production of aneuploid sperm. The discovery of factors associated with aneuploidy will be essential in learning how to prevent the deleterious effects that occur as a result of malsegregation of chromosomes.

By isolating RNA from mature nodules of the legume Lotus japonicas and employing an RT-PCR approach, two new cDNAs were characterized whose open reading frames code for members of the MIP (Major Intrinsic Protein) family of membrane proteins. These two genes products were termed (Lotus Intrinsic Membrane Protein 1 and 2) LIMP1 and LIMP2. Both LIMP1 and LIMP2 display all the hallmarks of the MIP protein family including, six putative transmembrane domains based on hydropathy plots, and the invariant NPA (asparagine-proline-alanine) signature motifs located symmetrically on two loops, which connect transmembrane helix 3-4 and transmembrane helix 5-6. Based on sequence information, LIMP1 is placed in the TIP (Tonoplast membrane Intrinsic Protein) subfamily of plant MIP proteins. Based on localization within the cells, TIPs are typically found on the tonoplast membrane of plant vacuoles. Functional characterization of LIMP1 by expression in Xenopus oocytes has demonstrated that it is a waterselective aquaporin. Using Northern blot techniques combined with immunolocalization, the expression patterns for LIMP1 were analyzed. The results indicate that LIMP1 is expressed predominately in nodules and roots. Based on Western blot analysis, however the highest LIMP1 protein levels are found in roots. LIMP1 appears to be regulated by diurnal cycling conditions, with expression increasing during the day hours, but falling off at the beginning of the night cycle. Increases in LIMP1 expression precedes the time of the diurnal cycle when the plant rate is the highest. This finding suggests that LIMP1, similar to other aquaporins, is regulated in response to the need for trancellular water uptake and transport from the soil during times of high transpiration stream activity. Although the LIMP2 protein shows some sequence similarity to LIMP1, it displays several features which distinguish it. Based on primary sequence identities, LIMP2 is classified in the plant NIP (Nodulin-like Intrinsic Protein) subfamily of MIP proteins which takes its name from the patriarch protein, nodulin 26. Nodulin 26 is a soybean nodule protein localized exclusively to the symbiosome membrane which surrounds the symbiotic bacteria during the nitrogen fixation process. Comparison of the LIMP2 protein sequence with other MIP family members shows that it shares the highest sequence identity (68%) with nodulin 26. In addition to structural similarity, nodulin 26 and LIMP2 also share the ability to facilitate the transport of water as well as small uncharged solutes (e.g. glycerol) upon expression in Xenopus oocytes. Analysis of the tissue distribution of LIMP2 by Northern blot and immunolocalization reveal that the LIMP2 transcript is expressed only in nodule tissue and that this protein, like nodulin 26, is present on the symbiosome membrane. LIMP2 and LIMP1 both show the ability to transport water, but the rate of transport is several-fold lower than that observed for high water transport MIP proteins such as mammalian aquaporin 1 (AQP1). Based on the crystal structures of AQP1 and the bacterial glycerol facilitator protein (GlpF), models of the selectivity filter for LIMP1 and LIMP2 were generated and analyzed in order to resolve the underlying characteristics for these observed differences. The proposed selectivity filter for LIMP2 displays a pore large enough to accommodate the passage of glycerol. Further, three key residues, a tryptophan in helix 2 and a valine in helix 5 and an arginine in the second NPA loop, form an amphipathic signature similar to the selectivity filter of the glyceroporin GlpF and are proposed to account for the glycerol permeability of nodulin 26 and LIMP2. In silica modeling of the predicted pore for LIMP1/TIPs shows a unique set of features. It shows a higher hyrdrophobic character than AQP1 and unusual substitutions, a histidine in helix 2 which replaces an invariant phenylalanine, an isoleucine in helix 5 which substitutes for a histidine, and a valine in the second NPA loop which substitutes for an invariant arginine. These substitutions suggest that the pore structure of LIMP1 is quite unlike the two models (AQP1 and GlpF), and further insight into factors that contribute to its rate and selectivity will have to await more detailed structural analysis. In addition to sequence and functional similarities, both nodulin 26 and LIMP2 share a conserved phosphorylation motif for CDPK (Calcium Dependent Protein Kinase) located on the cytosolic carboxyl terminal tail. In vitro assays using the carboxyl terminal sequence of LIMP2 suggest that similar to nodulin 26, it is phosphorylated by CDPK and is likely a target for calcium-dependent phosphorylation. Using an antibody which specifically recognizes the phosphorylated form of nodulin 26, a developmental pattern of phosphorylation is observed, where immature nodules show no or very low levels of phosphorylation and very old nodules show no phosphorylation of nodulin 26 although the protein is present in both young and old nodules at the same level as in mature nodules. Using this phosphorylation-specific antibody it was also determined that in mature soybean nodules the inherent level of phosphorylation is stimulated in response to osmotic stress (drought and salinity). Taken together, these results show that Lotus japonicus nodules have two MIP proteins. LIMP1 is a member of the TIP family of plant MIP proteins which is found predominately in root tissues where it forms a water selective channel, and its expression is controlled by diurnal variation. Its role in roots and nodules remains unknown, but it could play a role in transcellular water flow and cell volume regulation as proposed for other TIPs. In contrast, LIMP2 appears to be the Lotus japonicus ortholog of nodulin 26, an aquaglyceroporin that is localized on the symbiosome. LIMP2 and nodulin 26 are phosphorylated and likely regulated by calcium signaling through a symbiosome membrane CDPK. Previous studies show that phosphorylation of nodulin 26 enhances its water permeability. The present work suggests that phosphorylation by a calcium-dependent kinase is stress regulated and that phosphorylation of nodulin 26 may be among the osmoregulatorry responses of nodules necessary for stress adaptation, possibly by facilitating rapid water/solute flow to buffer the infected cell cytosol and symbiosome to fluctuations in osmotic gradients.

Problem Eating disorders, in the form of anorexia nervosa and bulimia nervosa, have been recognized as significant mental-health issues for the last three decades, and the incidence is rising as we approach the new millennium. Currently, many women who come into a mental-health setting due to depression, anxiety, low self-esteem, relationship issues, sexual issues, etc., are also struggling with eating-disordered behaviors, thoughts, and feelings. These behaviors, thoughts, and feelings may remain well hidden from the counselor throughout the course of therapy or until they become severe and more difficult to treat. Because eating disorders are very complex involving psychological, physical, and mental functioning, and because the symptoms become progressively more severe, early detection and intervention are essential for optimal outcome. Numerous assessment instruments exist but are not employed until obvious signs of eating disorders are exhibited. This study was designed to develop a subscale from the MMPI-2 items which will screen for eating disorders. Since the MMPI-2 is widely used early in the process of psychological evaluation, it was deemed the desirable instrument to use. Method The methodology for this study involved scale development and included four phases. Phase 1 was the initial study in which 354 MMPI answer sheets from eating-disordered individuals were compared to 238 MMPI answer sheets from non-eating-disordered individuals in order to determine MMPI items which differentiate between the two groups. Phase 2 utilized expert judges to evaluate the pertinence of each item on an eating-disorder questionnaire and to assign directionality. Phase 3 involved administering the items remaining after the first two phases to a new research sample comprised of eating-disordered and non-eating-disordered subjects. Phase 4 entailed eliminating the items which did not meet the total correlation criterion, and computing internal consistency using Cronbach’s alpha coefficient for the remaining items. Results This research resulted in the development of a 68-item proposed MMPI-2 subscale to screen for anorexia nervosa and bulimia nervosa. Each item met the differential criterion at the .01 level and the correlation coefficient at .33. The proposed MMPI-2 subscale has a reliability of .971 and its composition is unifactorial. Conclusions This research establishes the efficacy of utilizing the MMPI-2 to screen for eating disorders. Additional administration of the instrument is needed before it should move from research into practice.

Summary The consideration of the medical importance of phenothiazine derivatives and the known cancer chemotherapeutic value of the nitrogen mustards made the synthesis of some nitrogen mustard derivatives of the phenothiazines of particular interest. In the course of this research the N-{2-[bis-(2-chloroethyl)amino]ethyl} derivatives of phenothiazine, 7H-benzo[c]phenothiazine, 7H-dibenzo[c, h] phenothiazine, and 14H-dibenzo[a, h] phenothiazine were synthesized for anticancer evaluation. Also submitted to the Cancer Chemotherapy National Service Center for testing were the N-(2-chloroethyl) monofunctional nitrogen mustard derivatives of the first three phenothiazines mentioned above.