Insulin secretion from the pancreatic islet β cell is required for normal glucose homeostasis. The pancreatic islet has an extensive capillary network lined by endothelial cells. These endothelial cells are known to produce factors that can enhance β cell survival and proliferation. My hypothesis is that the islet endothelial cell also produces paracrine factors that can impact β-cell secretory function. The aim of my study is to determine the time course over which secreted factors from islet endothelial cells can enhance glucose stimulated insulin secretion (GSIS) from islet β cells. To perform these studies, we first generate islet endothelial cell conditioned media (IECCM) from MS1 cells, a transformed murine islet endothelial cell line. Pancreatic islets are then exposed to IECCM for up to 48 hours, and insulin release in response to basal (2.8 mM) and stimulatory glucose levels (20 mM) is determined. We have shown that exposure of islets to IECCM for 48 hours does not alter basal insulin release, but enhances glucose-stimulated insulin release by 50%. My ongoing studies will determine whether exposure of islets to IECCM for 1, 4, 12 or 24 hours is also able to enhance glucose-stimulated insulin release. These time course experiments will form the basis for studies planned for my senior year, where we will determine whether this increase in insulin release results from increased insulin biosynthesis, or is independent of insulin production, acting for example to stimulate exocytosis of preformed insulin-containing secretory granules. These studies may uncover the existence of a novel endothelial-endocrine axis that is crucial for β-cell function. These data could lead to new targets for therapeutic drugs that promote release of endothelial factors that regulate β-cell function.

In Acute Myeloid Leukemia (AML), adhesion of leukemia cells to the bone marrow microenvironment confers resistance to chemotherapy drugs. The receptors CXCR4 and CD44, an E-selectin ligand when properly glycosylated, have been implicated in the poor prognosis of AML. It has been shown that adhesion inhibitors restore chemotherapy sensitivity, but the E-selectin pathway has not been previously investigated. Previous data have shown that 81% of the AML patients tested showed at least 10% expression of E-selectin ligand. Hence, we proceeded to test E-selectin by utilizing antibodies to E-selectin ligand and chemokine receptor CXCR4 to block the association with stromal cell components in vitro. CFSE staining protocol was then utilized to read fluorescence intensity of cells with and without antibodies. The ability of HECA 452, antibody to E selectin ligand, and GMI, the bifunctional inhibitor to E-selectin and CXCR4, to block adhesion to stromal cell line HS27a was investigated. To further assess the interaction between E-selectin and AML cells, plates coated with E-selectin as opposed to HS27a stromal cells were utilized in the blocking assay. It was found via the use of Cell Titer Glo that HECA and GMI displayed a significantly lower luminescence compared to their respective control indicating that both antibodies were effective in preventing AML cell adhesion. We plan to continue this study by utilizing chemotherapeutic agents Cytarabine and Daunorubicin in conjunction with the antibodies to eliminate AML cells in vitro. Our research will allow us to determine how to prevent chemotherapy resistance which would make it effective in treatment of patients with Acute Myeloid Leukemia.

Most adults have some degree of atherosclerosis. In a subset of these individuals, atherosclerosis progresses more rapidly. Increased activity of proteases (enzymes that digest proteins) in atherosclerotic plaques is a plausible cause both of atherosclerosis progression and of atherosclerotic plaque rupture. Plaque rupture is often a terminal event that results in blockage of arteries, heart attacks and strokes. Urokinase plasminogen activator (uPA, a protease that is present in plaques) functions by converting plasminogen (a zymogen, or enzyme precursor) into its active form, plasmin. The uPA/plasminogen pathway facilitates macrophage migration, enhances the release of inflammatory mediators, and activates other proteolytic enzymes, all of which may contribute to atherosclerosis progression and plaque rupture. We hypothesize that inhibition of uPA will decrease atherosclerosis progression and prevent plaque rupture. To begin to test our hypothesis, we use a mouse model of uPA-accelerated atherosclerosis (mice that are genetically predisposed to develop atherosclerosis and express high levels of uPA). We inject an antibody (mU1) that binds to and inhibits uPA, or a control antibody, and we measure atherosclerosis in aortic roots and pinned aortas. We anticipate finding less atherosclerosis in mice treated with mU1. We also expect to find fewer macrophages in atherosclerotic plaques of mice treated with mU1. If we find that inhibition of uPA decreases atherosclerosis progression and that it is also well tolerated by mice, we will later set up an experiment in older mice to determine whether anti-uPA treatment with mU1 will prevent plaque rupture. If uPA activity contributes to atherosclerosis progression, and if inhibiting uPA decreases atherosclerosis progression and prevents plaque rupture, drugs could be developed that inhibit uPA. These drugs could be used to treat humans who are at high risk for atherosclerosis progression and plaque rupture.

DNA benefits from a system of self-maintenance, containing genes that code for the repair of its damage. The base-excision repair pathway serves as the primary means of preventing mutations attributed to oxidative stress, such as 8-oxoguanine (8-oxoG), within both the nuclear and mitochondrial DNA. Initiated by the 8-oxogunine DNA glycosylase (OGG1), the removal of 8-oxoG is known to prevent genomic instability and neoplasia. The current study presents new data indicating a role for OGG1 in the maintenance of energy homeostasis at both cellular and whole-body levels. Oxidative stress introduced via high-fat diet (HFD) yielded increased adiposity and hepatic steatosis within the OGG1 knockout (OGG1-/-) mouse model. The HFD-fed OGG1-/- mice were also prone to increased levels of plasma insulin and impaired glucose tolerance. The same mice exhibited a higher resting VCO₂, indicating a preference for carbohydrate metabolism versus fat oxidation. Analysis of expression levels in the livers of OGG1-/- mice revealed significant downregulation of the metabolism-regulating transcriptional coactivator PGC1 and other genes integral to fatty acid oxidation. Consistent with a reduction in PGC1a, multiple genes involved in the TCA cycle were significantly downregulated. Also noted was a reduction in glycogen stores within the OGG1-/- livers, and fasting plasma ketones were significantly reduced. We conclude that OGG1 deficiency alters cellular substrate metabolism, fosters a fat-sparing phenotype, and renders the OGG1-/- mouse highly susceptible to obesity and related diseases. An understanding of OGG1 and its role in energy homeostasis will contribute to the direction of research within the realm of disease initiation due to oxidative stress.

Comic books have long been used as an educational tool to improve and protect public health in the U.S. In 2008, the Native Comic Book Project (NCBP) was launched as a youth-focused community education project of Native People for Cancer Control (NPCC); a National Cancer Institute funded Community Network Program Center, based at the University of Washington. The availability of cancer screening tests, high quality health care, encouragement for health promoting behaviors, and access to cancer clinical trials, are often not available to tribal communities. As a result, American Indians and Alaska Natives (AI/ANs) experience the worst cancer related disparities and have the poorest survival rate among all other racial/ethnic groups. The purpose of the NCBP is to reduce cancer risk by involving AI/AN youth in creating comic books on health topic such as health living, tobacco use, cancer risk, disease prevention, and physical fitness. The NCBP is modeled after Dr. Michael Bitz’s The Comic Book Project, but is adapted for AI/AN youth and incorporates Native storytelling and traditional ways of living. We are currently conducting a formal evaluation of the NCBP using pre- and post-intervention assessments to measure knowledge, habits, and decision-making regarding tobacco use, healthy eating, exercise habits, and human papilloma virus (HPV).

While red blood cell transfusions require basic blood type matching (A+, B-, O+, etc...), effective platelet transfusions require complex histocompatibility matching, testing to make sure specific alleles of genes are the same between donor and recipient so as to prevent rejection; this is similar to the testing required for organ transplants. Currently, platelets are transfused from random donors until a patient shows signs of rejection, at which point a blood center must attempt to find a histocompatible donor out of its registry to donate on short notice. Using a canine model, the Slichter lab has created a treatment, through a combination of filtration leukoreduction to remove antigen presenting white cells (APCs) and irradiation to inactivate residual APCs, which may prevent transfused recipients from rejecting non-histocompatible donor platelets. In my research, I will focus on canine immune responses to platelet transfusions. The lab transfuses canines with mismatched, treated platelets and measures the recovery and survival of radiolabeled donor platelets to document rejection; antibody levels are also measured, to correlate decreased donor platelet recoveries and survivals as being immune mediated. I perform an Enzyme-Linked Immunoabsorbent spot (ELI-spot) assay to test the production of gamma interferon (IFN-g). IFN-g is produced primarily by activated T-h1 and CD8 lymphocytes as well as other cells such as macrophages and mast cells. As such, the ELI-spot serves as a direct indication of the very beginning of an immune response. My research aims to expand on the current antibody data and directly correlate the recipient’s response to transfused platelets with the results of the ELI-spot assay. This research allows a direct link between cell interactions and immune responses, which can then be compared to the donor’s platelet recovery and survival data to see if a correlation exists. This will allow us to better judge the effectiveness of various platelet treatments by identifying significant correlations between in vitro data and post-transfusion platelet responses. Not only can this lead to breakthroughs in platelet delivery, but the results may also be useful in detecting early organ transplant rejection.

Schizophrenia is a debilitating neuropsychiatric disorder with a worldwide prevalence of 1%. Although the heritability of schizophrenia is high, there is a large proportion of affected individuals with no family history. We are testing the hypothesis that many different severe mutations are the causal genetic factors for schizophrenia. In order to test our hypothesis, we sequenced the exomes of approximately 50 trios consisting of an affected individual and his/her parents who are free of mental illness. Exome sequencing is a rapid and robust method for looking at only coding portions of the genome, and allows for the detection of rare variants in those regions. Sequence data is then run through several bioinformatics filters to ensure that the variant is not common in the general population, and that the variant is probably damaging to the gene product. All variants that pass through the bioinformatics filters are validated with Sanger sequencing. De novo mutations were found in 19 different genes in 11 trios. Of the mutations 13 were single base pair changes, and 6 were truncating nonsense or insertion/deletion events. After these experiments have been carried out on approximately 100 trios, we will use the group of candidate genes to identify critical pathways that may be affected in schizophrenia. Understanding the structure of these pathways may allow for the development of targeted therapies for schizophrenia.

Schizophrenia is a devastating neurodevelopmental disorder whose genetic influences continue to be elusive. Rare, gene-disrupting genomic deletions and duplications – called copy number variants (CNVs) – have been implicated in schizophrenia; however, much remains to be understood about which genes are causative, as well as the cellular mechanisms involved. Biological follow up of individual CNVs may give insight to the origin of schizophrenia. We focused on a CNV present in a patient with early onset schizophrenia. This mutation duplicates the 5' ends of two genes that lie head-to-head on chromosome 11q22: DCUN1D5, a previously uncharacterized gene predicted to be involved in cullin neddylation of ubiquitin ligase complexes, and DYNC2H1, a dynein active in cilia. Both are expressed in brain and are plausible candidate genes for schizophrenia. Using RNAseq, we detected novel DCUN1D5 transcripts in the patient’s lymphoblasts that we predict will result in aberrant, truncated DCUN1D5 protein. To test the conformational stability of these aberrant proteins, we then expressed epitope-tagged full length and truncated DCUN1D5 in HEK293 cell lines. These transfected cells also allow us to study the functional capabilities of the aberrant proteins in the cullin neddylation pathway, and the specific cullin binding partners of DCUN1D5. Investigation of these aberrant proteins and their function may shed light on pathways that contribute to schizophrenia, potentially guiding the search for new candidate genes and the development of novel treatment strategies.

Human embryonic stem cells (hESCs) are pluripotent, or able to differentiate into all cell types of a developing embryo. They can be used to engineer human tissue, but are prone to immune rejection when used in medical treatment. Similar to hESCs but derived from the reprogramming of adult somatic cells, human induced pluripotent stem cells (hiPSCs) were developed that have the potential for use in regenerative therapy with less risk of rejection. However, hiPSC research faces its own challenges, such as inefficient reprogramming rates and variation in observed pluripotency. Previous research has shown that methylation patterns of hESCs can be distinguished from hiPSCs at both the DNA and histone level, suggesting less pluripotency in hiPSCs than in hESCs. While exhibiting greater overall methylation on DNA but less on histones, many of the hypermethylated DNA regions of hESCs are adjacent to these differentially methylated histones. The location of the hypermethylated regions suggests that erasing specific histone methylation marks may improve hiPSC reprogramming rates by creating a more open chromatin landscape, and may create a cascade effect that allows for additional methylation of the genome. This experiment investigates the correlation between histone methylation and hiPSC reprogramming efficiency and accuracy. We reprogrammed human fibroblast cells with lentiviruses expressing various demethylases and transcription factors in order to compare resultant hiPSC methylomes to traditional hiPSC and hESC lines. Opening of the chromatin is expected to improve reprogramming rates and allow for proper methylation of the genome, creating an hiPSC closer in phenotype to hESCs and more useful for clinical therapies.

Toxoplasma gondii (T. gondii), the parasite responsible for human Toxoplasmosis, has previously been shown to be inhibited by Bumped Kinase Inhibitors (BKIs) via inhibition of Calcium Dependent Protein Kinase 1 (CDPK1) in vivo. While BKIs inhibit T. gondii very well with high selectivity for parasite and not human functions, and have low toxicity, CDPK1 itself is significantly less inhibited by the compounds in vitro, suggesting that the observed inhibition may in part be due to other enzymes. Using genomic data, we selected three T. gondii kinases with similar gatekeeper regions to CDPK1 that could be structurally vulnerable to BKIs as well. In an effort to identify and understand these Off-target enzymes, two ongoing experiments were conducted. The first aim was to purify the suspected Off-target enzymes and assay them in vitro against the same BKI compounds that were so effective against CDPK1. The second aim was to genetically manipulate the protein structure of the Off-targets in the gatekeeper region of the enzyme to match that of the known CDPK1 enzyme, to verify the mechanism of action of the BKIs. The likely enzymes to be Off-targets were difficult to purify, and the project is in progress. Future work depends on new methods to purify the Off-targets, in order to make progress toward completing the goals of the project. CDPK1 is an attractive target for therapeutic development for Toxoplasmosis, and further knowledge of Off-target enzymes that are vulnerable in the same way in T. gondii can lead to much-needed low resistance therapeutics.

 Mycobacterium tuberculosis infection is a leading cause of death worldwide, but the full details of how the human immune system responds to these bacteria are not known. Our lab studies CD1 proteins which allow T-cells to respond to bacterial lipids. We discovered that low expression of CD1A on dendritic cells is associated with an increased susceptibility to tuberculosis; however, the molecular details of CD1A-deficiency are not known. I cloned and analyzed the CD1A promoter from four CD1A-deficient and five control individuals. I identified twelve genetic variations in the CD1A promoter. Of these, eight were single nucleotide polymorphisms (SNPs), three were insertions or deletions, and one was a variable tandem repeat. One SNP (-199 T/C) and the variable tandem repeat showed an association with CD1A-deficiency. A T/T genotype at position -199 from the transcription start site correctly identified five out of five control subjects while a C/C genotype identified three out of four CD1A-deficient subjects. The variable tandem repeat was a string of 20 to 60 thymine bases. The CD1A-deficient subjects showed uniform repeat lengths, while the repeat lengths among control subjects were variable. These data identify two loci within the CD1A promoter that will be the target of future studies aimed at understanding the molecular mechanism of CD1A-deficiency.

South Africa’s KwaZulu-Natal province currently has the highest HIV prevalence in the world at approximately 30%. The University of Washington’s International Clinical Research Center is currently conducting a pilot program for a new HIV testing method, Home-Based Counseling and Testing for HIV (HBCT), to address the HIV epidemic. A mathematical model of heterosexual HIV transmission in KwaZulu-Natal was created to evaluate the long-term consequences of HBCT on HIV prevalence. This model simulates a population of susceptible and infected individuals with varying sexual activity levels and therefore, risk. The model also divides the population among twelve age cohorts and the infected individuals are further divided into three HIV stages and a final AIDS stage, as defined by the World Health Organization. The model was created in MATLAB and validated with HIV prevalence data for KwaZulu-Natal from the Joint United Nations Programme on HIV/AIDS. Preliminary results from the model indicate that HBCT will diagnose more cases of HIV than other testing methods and, with successful linkage to care, will decrease HIV prevalence in the future. While the possibility of HIV elimination by HBCT is yet to be determined, preliminary results indicate that HBCT is effective at linking infected individuals with care, including antiretroviral therapy. This work has the potential to guide health policy in South Africa, and given the newfound effectiveness of pre-exposure prophylaxis and treatment as prevention, HBCT provides a promising future for controlling the HIV epidemic.

The nature of Systemic Lupus Erythematosus (SLE) involves a complex network of uncontrolled activations of signals, which leads to a vast range of symptoms. SLE is an autoimmune disease that manifests itself when the immune system attacks the body’s cells and tissues, resulting in inflammations in joints, organs, and skin. The course of the disease is unpredictable and much more is to be understood about the mechanisms of the illness and the background of the patients who are prone to the disease. This study explores both the mechanistic and clinical aspect of SLE in an attempt to gain a broader view of the disease. Using 3 interferon stimulated genes, MX1, PKR, and CXCL10, we evaluated interferon scores of 66 patients. We then utilized cluster analysis to divide patients into two main groups based on interferon scores: Group 1 consisted of controls and SLE patients with low interferon scores (0.4), Group 2 consisted of SLE patients with high interferon scores (10.4). These groups were compared with interferon stimulated genes, RNase1 and Toll-Like Receptors 7 (TLR7), which positively correlated with interferon scores. A system called Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) is often utilized to quantitatively assess patients’ disease activity. The criteria in the evaluation of patients included symptoms often associated with SLE. The score in SLEDAI reflected the disease activity of the patient. In this study, we investigated the relationship between SLEDAI and interferon score, in which we found no correlation. However, when we examined Group 2, there was a trend towards better correlation with SLEDAI. In the clinical aspect, we examined age, race, and gender of SLE patients. In addition, we collected information on autoantibody titers such as double stranded DNA (dsDNA), antiphospholipid antibodies, and antinuclear antibodies (ANA), all of which may be important indicators of disease activity.

Acute Myeloid Leukemia (AML) is a malignancy of blood cells due to the proliferation of undifferentiated cells leading to hematopoietic insufficiency. It is known that genetic changes are causally associated with AML. FLT3 gene (Fms-Like Tyrosine kinase 3) is essential in regulating normal hematopoiesis, and mutations in these genes have been shown to be common in AML and lead to autonomous cell growth. In addition to mutations, our lab has identified novel transcript variants in FLT3 gene that may have biologic and clinical significance. These novel variants are a result of alternative splicing of exons, a regulatory developmental process that can result in production of different proteins from the same gene. My project is to study these novel FLT3 variants via molecular techniques and to determine the prevalence of these transcript variants from a cohort of patients with AML. I tested for the presence of FLT3 transcript variants in AML patient specimens relative to normal bone marrow. I analyzed my amplified products with a software called Gene Mapper, which identifies variations in size and expression rate of gene transcripts. In the normal bone marrow control, only one (wild type) product was identified. However, in the leukemic specimens, 4 different transcript variants were identified at different frequencies. The most intriguing transcript was missing exons 19 and 20. This would cause the tyrosine kinase receptor to have a truncation of the functional, kinase domain of the receptor. This variant would lead to the translation of a protein product that is devoid of kinase activity. I plan to correlate our data with clinical outcome to determine the clinical significance of these transcript variants. If associated with poor outcome, patients can get placed into a higher risk category, where they can be treated with more intensive chemotherapy followed by bone marrow transplantation.

HIV-2 is endemic in West Africa, but has a much more limited spread worldwide than HIV-1. Compared to HIV-1 infection, HIV-2 is characterized by a much longer asymptomatic stage, lower plasma viral loads, slower decrease in CD4 cell count, decreased AIDS associated mortality rate, and lower rates of genital tract shedding, mother to child transmission, and sexual transmission. Nonetheless, a signficant proportion of HIV-2 infected individuals will eventually progress to AIDS, and many of these individuals will benefit from antiretroviral therapy (ART). However HIV-2 is intrinsically resistant to nonnucleoside reverse-transcriptase inhibitors (NNRTI's) and the fusion inhibitor T-20, and reports suggest HIV-2 may be partially resistant to protease inhibitors. However the mutations in HIV-2 that confer resistance are not well characterized. Improving treatment of infected individuals requires a thorough understanding of the mutations that occur in the HIV-2 strains of ART treated patients and the effect of these mutations on the efficacy of ART. Plasma samples used in this study were obtained from ARV-treated HIV-2 infected individuals participating in the Senegalese Initiative for Access to Antiretrovirals (ISAARV) program. A phylogenetic analysis approach was used to identify common mutations occuring in the HIV-2 strains of individuals receiving protease inhibitor (PI) based therapy. Gaining an understanding of the mutations that occur in the HIV-2 strains of indiviuals receiving PI-based ART and their effect on ART efficacy is a first step in improving treatment of HIV-2 infected individuals to prevent progression to AIDS.

Syphilis is a disease that is increasing in the United States. It is estimated that, every year, twelve million new syphilis infections occur globally. The development of a vaccine is therefore a high priority. Proteins in the outer membrane of the bacterium that causes syphilis, Treponema pallidum subsp.pallidum (T. pallidum) and surface-exposed epitopes that are recognized by the immune system must be identified to be used as targets for vaccines and new serological tests. One promising target for vaccine development is TprC, a predicted outer membrane protein with surface-exposed loops. This project will determine whether these predicted surface loops on the TprC protein are recognized by antibodies. TprC also varies among syphilis strains in discrete regions of the protein. The protein TprC was synthesized as small peptide fragments consisting of 20 amino acids, with 10 amino acids of overlap between each peptide. All of the peptides fragments including peptides of variable regions will be tested using anti serum from each strain. These peptides are exposed to antibodies from infected rabbit sera and visualized by a labeled secondary antibody. Those peptides visualized by the secondary antibody are regions of TprC recognized by the immune system. These ongoing experiments will determine whether the surface-exposed regions of Tpr C are the major targets of the antibody response and whether antibodies induced by infection of one strain will be effective against another strain; both crucial in vaccine design.

Cell labeling and tracking hold enormous potential to illuminate cellular pathway mechanisms. Because current labeling methods are either unsafe or unavailable for humans, a need exists for a safe and effective cell labeling technique suitable to track cells administered for cell therapy. The stable carbon isotope, C13, is safe for human use, incorporates into cellular DNA when taken up as C13 glucose in culture medium, and can be detected by GC mass spectrometry. We proposed to evaluate the use of C13 to label and track exogenous bone marrow derived macrophages (BMMCs) administered after myocardial infarct (MI). To confirm exogenous BMMCs can be detected in the heart, we injected green fluorescent protein (GFP) labeled BMMCs into the tail vein of mice three days following MI. Some heart sections were assessed for GFP positive cells by immunohistochemistry. Other hearts underwent dispersion and isolation of macrophages. Quantitation revealed the percentage of exogenously delivered C13 labeled GFP BMMCs in the cardiac macrophage population was 10 percent. Preliminary cell proliferation assay results suggest no considerable difference between the growth of BMMCs with or without C13 glucose. Finally, we cultured GFP BMMCs for ten days in 100% C13 glucose media for tail vein injections to mice following MI. Three days later, we excised the hearts to isolate cardiac macrophages. Cultured heart cells and macrophages were analyzed via GC mass spectrometer to quantify labeled DNA. Although we could detect C13 in in vitro labeled macrophages, increased C13 cellular label could not be detected in isolated macrophages. Unlabeled macrophages appropriately had no increase in label. Ultimately, while this study was unsuccessful in detecting a C13 cellular label in vivo with a GC mass spectrometer, in vitro successes suggest technique refinement could lead to a powerful and safe therapeutic stem cell label.

In Acute Myeloid Leukemia (AML) the adhesion interaction between the AML blast cells and the bone marrow microenvironment confers resistance of the leukemia cells to the chemotherapy drugs. One possible receptor that might facilitate this adhesion is the VLA-6 integrin. Our laboratory has been found it to be highly expressed in the AML stem cells, the cells that are believed to give rise to leukemic relapse. To investigate the role of the VLA-6 receptor, I performed assays to observe the effects of blocking this receptor on cell adhesion as well as the chemotherapy resistance. The adhesion assay consists of measuring AML patient cell binding to laminin-coated wells, as laminin is the ligand for the VLA-6 adhesion receptor. Adhesion mediated by VLA-6 is assessed by measuring binding in the presence or absence of the anti-VLA-6 antibody. The chemotherapy resistance assay involved the same conditions as the adhesion assay. We expected that blocking the VLA- 6 receptor would prevent the cells from adhering to the laminin and that this would also lead to lessened drug resistance. However, for the cells from one patient studied so far, there has not been a significant difference between the amount of adhered cells in these conditions. Additional experiments are in progress with more patient samples to elucidate the significance of the VLA-6 receptor in AML physiology and survival.

Embryonic diapause or delayed implantation is an occurrence that effectively lengthens the gestation period of the embryo and allows offspring to be born into optimal conditions. In response to adverse environmental conditions, the mouse blastocyst enters a reversible state of cell quiescence, or extremely slowed development, resulting in delayed implantation. Our aim is to uncover the mechanisms that occur in diapause that allow stem cells to endure stressful environments. Theoretically we could generate “super” stem cells that retain pluripotency for longer periods and withstand harsh in vitro treatments. Gene expression profiling of the stem cell compartment of diapause embryos showed that the transcription factor Runx1 is upregulated and might be required to initiate diapause. Runx1 thus far has shown to induce a state of cell quiescence in haematopoietic stem cells (HSCs) and is responsible for haematopoiesis, the formation of blood cellular components, in early development1. Runx1 is also known as acute myeloid leukemia 1 protein (AML1) because mutations in it are the most common aberrations found in acute leukemia2. Additionally, we observed up-regulation of RUNX1 protein expression level in the primitive endoderm (PE) layer of the diapause embryos. We hypothesize that the increased expression of RUNX1 is required to maintain pluripotency of the stem cell compartments in the embryos. In vivo mouse models of Runx1 wild-type (+/+), heterozygous (+/-), and deficient (-/-) diapause embryos will be examined using confocal microscopy with antibodies against RUNX1, pluripotency marker (OCT4), and PE marker (GATA4), and the resulting morphologies of the Runx1 embryos will also be characterized.

The Institute of Medicine‘s (IOM) report on health disparities, “Unequal Treatment”, identified the medical system’s inability to distinguish minority groups with meaningful specificity as a primary barrier to recognizing critical disparities in health care and outcomes. A recent follow-up report on “Race, Ethnicity, and Language Data” called for more detailed demographic data collection. This study evaluated the validity of self-reported primary language to detect disparities in diabetes care across language groups of first generation, mostly non-English speaking outpatients at an international clinic. The REALL (Race, Ethnicity and Language proficiency Level) study is a cross-sectional pilot study, surveying a convenience sample of immigrant and refugee patients receiving primary care at an urban, county hospital. Participants were eligible for this study if they had diabetes based on medical record data and completed the REALL survey question about how many and what languages they speak, their reading ability in those languages, and the culture they most identify with. Survey data were linked with medical record data that included hemoglobin A1C (HA1C), low density lipoprotein (LDL) and systolic blood pressure (SBP). Diabetes control was evaluated across the 6 largest language groups. 250 patients were eligible for the study. The survey data were 90.84% concordant with registration data for primary language. Mean HA1Cs were lowest among Vietnamese patients, significantly lower than Somali patients and Cambodian patients. While 52% of Vietnamese patients had HA1C<7, only 26% of Somali patients had HA1C<7. There were no statistically significant differences across language groups in mean LDL level or SBP. Results suggest that there is variability in diabetic outcomes across language groups in first generation immigrant communities. Recent refugee communities are potentially more stressed due to lack of acculturation and community establishment. Further inquiry into their duration in the US, the clinical care system, and their dietary practices are advised.