Mast cells are critical effectors of allergic disease that can be activated by numerous stimuli. We have examined mast cell control by the inflammatory cytokine, IL-33, as well as IgG. In the first study reported here, we found that the synthetic glucocorticoid, dexamethasone, potently and rapidly suppressed IL-33-induced cytokine production from murine bone marrow–derived and peritoneal mast cells, as well as human mast cells. Dexamethasone also antagonized IL-33-mediated enhancement of IgE-induced cytokine production and migration. Although dexamethasone had no effect on IL-33-induced phosphorylation of MAP kinases or NFκB p65 subunit, it antagonized AP-1 and NFκB-mediated transcriptional activity. Finally, intraperitoneal administration of dexamethasone completely abrogated IL-33-mediated peritoneal neutrophil recruitment and prevented plasma IL-6 elevation. These data demonstrate that steroid therapy may be an effective means of antagonizing the effects of IL-33 on mast cells in vitro and in vivo, acting partly by suppressing IL-33-induced NFκB and AP-1 activity. In the second study reported here, we found that Fcγ receptor crosslinkage activated the transcription factor Stat5B through a Fyn kinase-dependent pathway. We then showed that STAT5B is critical for IgG-induced cytokine production by mast cells but not macrophages. To expand these studies, we employed the K/BxN model of inflammatory arthritis, which has roles for mast cells and macrophages. In this model, Fyn or STAT5B deficiency only affected the arthritic flare that primarily depends on mast cell degranulation, without affecting the severity of the joint swelling. By contrast, Lyn kinase deficiency significantly exacerbated arthritis. These studies indicate a clinically relevant, lineage-restricted role for the Lyn/Fyn-STAT5 cascade. Collectively, our work demonstrates that mast cell activation by diverse stimuli can be suppressed by steroid intervention and selectively targeted by disrupting kinase-transcription factor pathways.

Adult immunizations effectively reduce morbidity, mortality, and transmission rates of multiple diseases; however, outpatient providers often a struggle to convince patients to accept vaccinations. This project’s aim is to address vaccination rates in our adult population, focusing first on the influenza vaccine in year one (2016), and then on pneumococcal vaccine in year two (2017), by 1) using a strong quality improvement strategy (known as a Champion Team) and 2) implementing a clinic program consisting of provider training, improved documentation, and informative posters targeted at patients. A quality improvement strategy known as a “Champion Team” provided a strong mechanism through which we developed and implemented the interventions across both years. Specifically, the Champion Team consisted of key stakeholders (nurses, residents, physician faculty, and informatics expert) who identified, developed, and evaluated the program. Programming included an annual health care professional training session for each vaccine (early fall of 2016 and 2017 for flu, spring 2017 for pneumococcal), improved documentation strategies and nursing uptake, and informative posters in the clinic. We assayed data from our patient electronic health record to evaluate: the percentage of our patient population for whom an immunization was documented relative to the number of unique patients seen in our clinic during that time frame. This approach in year one showed a marked increase in influenza vaccination rates in our clinic. During the 2014/2015 and 2015/2016 flu seasons our clinic vaccination rates were 39.98% and 42.05% respectively. After implementation of our champion team and clinic wide program to increase rates in 2016 our vaccination rates for the 2016/2017 flu seasons was 50.88%. Pneumonia data for a full year are under analyses and will be included in this presentation. We anticipate a similar increase in rates for our pneumococcal vaccinations. Our Champion Team and clinic wide program were perceived as relatively low-effort interventions yet appeared to increase vaccinations over the course of this study. The replication of these findings across pneumonia data (pending) and, in future work, with the herpes zoster vaccine (planned for Year 3), will increase our confidence that increases in rates were attributable to these very accessible interventions.

This report describes the case of a 56-year-old man who presented with blurry vision, increased intraocular pressure, and conjunctival injection after posterior chamber intraocular lens implantation. Initially post-operative endophalmitis and foreign body inflammation were considered as differential diagnoses, but after further examination pan-uveitis was diagnosed. Uveitis is an ocular finding that may indicate several diseases, one of which is Behçet’s Disease. During the interview, the patient mentioned a history of apthous ulcers and genital ulcers which then lead to the clinical diagnosis of Behçet’s Disease. This report emphasizes that Behçet’s Disease is rare in Caucasians. Therefore, is frequently misdiagnosed in North America due to variable presentations and by not exploring the option when analyzing differential diagnoses. Early diagnosis and intervention will prevent the development of blindness and fatality due to complications of the disease.

The effects of alcohol abuse are multi-dimensional since alcohol is widely known to affect both the innate and adaptive immune systems. Recently, epigenetics has come into focus and has been implicated in many diseases as well as substance abuse disorders. Therefore, research efforts of understanding the epigenetic mechanisms underlying substance abuse effects including alcohol abuse have become more predominant. In our laboratory, we have studied different epigenetic changes induced by alcohol exposure including regulation of histone deacetylases (HDACs), histone quantity, and histone modifications such as acetylation and deacetylation. We have observed differential effects of acute and chronic alcohol exposure in human monocyte-derived dendritic cells (MDDCs) wherein our laboratory previously found that HDACs were modulated in MDDCs treated acutely with alcohol in vitro, and in MDDCs from alcohol users. Our previous work has also demonstrated that alcohol consumption affects the dendritic cell function by modulating inflammatory markers and cannabinoid receptors such as CB2 and GPR55 through epigenetic modifications. For instance, chronic alcohol exposure upregulates histone (H) 4 acetylation at lysine 12 (H4k12ac) and acute alcohol effects on histone acetylation are associated with an increase in GPR55 expression. The hypothesis of the study is that chronic alcohol modulates human MDDC function through epigenetic mechanisms. Therefore, the primary objective of this research project is to elucidate novel pathways involving histone post-translational modifications due to chronic alcohol exposure in human dendritic cells. For this study, monocytes isolated from commercially available human buffy coats were differentiated into MDDCs, which were treated with chronic alcohol levels of 0.1 % (100mg/dL) and 0.2 % (200mg/dL) for 5 days in the presence or absence of the histone acetyltransferase inhibitor NU9056 (50nM) or the GPR55 antagonist CID16020046 (5µM). Results showed that chronic alcohol levels upregulated H4K12ac in MDDCs and this was associated with a concomitant increase in GPR55 gene and protein expression. Further, NU9056 and CID16020046 were able to reduce alcohol-induced inflammatory chemokine MCP-2 and reactive oxygen species production indicating that H4K12ac may be an inflammation and oxidative stress regulator. Additionally, NU9056 and CID16020046 could potentially reduce alcohol-induced inflammation and serve as potential therapeutic targets for alcohol use disorders.

Vitiligo is an autoimmune disease of the skin characterized by epidermal depigmentation that results from CD8+ T cell-mediated destruction of pigment producing melanocytes. Vitiligo affects up to 1% of the population and current treatments are moderately effective at facilitating repigmentation by suppressing cutaneous autoimmune inflammation to promote melanocyte regeneration. In order to cause disease, CD8+ T cells must overwhelm the mechanisms of peripheral tolerance in the skin and if we understand the suppressive mechanisms that are compromised during vitiligo, we can potentially use this information to improve existing treatments or engineer novel interventions. Therefore, my goal is to characterize the regulatory factors in the skin that suppress depigmentation during vitiligo. Our lab has developed a mouse model of vitiligo that accurately reflects human disease and I used this model to demonstrate that regulatory T cells suppress CD8+ T cell-mediated depigmentation and interact with CD8+ T cells in the skin during vitiligo. In this model of disease, I investigated the molecules involved in regulatory T cell function and observed that the chemokine receptors CCR5 and CCR6 play different roles in regulatory T cell suppression. While CCR6 facilitates regulatory T cell migration to the skin, CCR5 is dispensable for migration but required for optimal regulatory T cell function. Additionally, I used our mouse model to demonstrate that Langerhans cells suppress the incidence of disease during vitiligo. Taken together the results from these studies provide novel insights into the mechanisms of suppression during vitiligo.

On June 8, 2006, Merck announced the debut of Gardasil, the world's first vaccine found successful in preventing human papillomavirus (HPV) infections, a sexually transmitted infection that is one of the main causes of certain cancers in men and women, including cervical, vulvar, penile and anal cancers. To promote the vaccine's release, Merck launched Gardasil's "One Less" advertising campaign that included television commercials, print ads and a consumerfocused website (www.Gardasil.com), each promoting the message that "you" could now be "one less woman" affected by cervical cancer ("One Less" campaign). The vaccine, tested and approved only for females age 9-26, was advertised to this age group, as well as parents or guardians responsible for making medical decisions for female minors. As the campaign launched, commercials depicted females laughing and enjoying hobbies while mentioning the positive decision they made to receive the Gardasil vaccine. Many commercials also included portrayals of mothers talking happily about their decision to get their young daughters vaccinated. Interestingly, male figures were completely left out of Gardasil's "One Less" campaign ads, despite the fact that in reality, males administer the vaccine as medical professionals, transmit the infection as sexual partners, and suffer cancers as HPVinfected patients. Males were even left out of the ads as parents, who were always portrayed by women in the ad campaign. iv Informed consumers may have expected all this to change on Oct. 16, 2009 – three years after Gardasil's debut – when the Food & Drug Administration (FDA) approved the vaccine for use in males age 9-26 to protect against HPV-caused genital warts. Though Merck's vaccine was now accessible to more consumers than ever, the advertising that surrounded this medical breakthrough changed very little. Television commercials for the vaccine still promoted Gardasil primarily to women for the purpose of preventing HPV-related cervical cancer. Again, men were not featured in commercials as medical professionals, parents, guardians, romantic partners, or even as patients able to get the vaccine. Males did begin appearing on the vaccine's official website, however these depictions were limited to showing only young boys, who appeared standing with a mother's protective arm around them. Males that represent the older age range (up to age 26) were never shown. What effect does the lack of male representation have on the verbal and nonverbal message these ads are sending consumers about who fits in the target consumer group, as well as who is at risk for an HPV infection? On a broader level, how does gender representation as a whole affect pharmaceutical advertisements and the adoption of the potentially life-saving products they promote? How does a pharmaceutical technology become "gendered"? How do specific gender portrayals impact the educational aspects of pharmaceutical ads, which may shape a consumer's opinion of who is at risk for an illness, and who is responsible for its treatment or prevention? And how do these gender portrayals connect with, reflect or reinforce v dominant cultural beliefs about the roles males and females play in protecting themselves and others from disease? In this study, I investigate these questions using a blended cultural studies/social sciences research perspective, first looking at the controversial history of direct-toconsumer pharmaceutical advertising and the gender stereotypes that traditionally exist in this form of rhetoric. I then test the affect Merck's gender portrayals has on its ad message in a blind study done with a small sample population, which provides evidence that Merck's ads are confusing and exclusive of certain populations, particularly men. I then investigate how Merck's existing gender portrayals, and strong focus on women, reflect larger historical beliefs on the roles that males and females play in health care and in the family. I show how, through advertising, Gardasil has become "gendered" as a pharmaceutical technology for female children. From here, I will show how pharmaceutical companies, such as Merck, have both reflected and reinforced the belief that women are the primary caregivers to children, how this stereotype is both damaging and statistically incorrect, and how using it targets Gardasil ads to a very narrow population of consumers, miscommunicating the message of who is at risk for illness contraction and perhaps even damaging sales in addition to prevention. I later provide evidence that Merck's current Gardasil ad series and other actions in the marketplace are dangerously misleading certain populations regarding the nature of the HPV virus, the protective abilities of the vaccine, and the populations responsible for accessing Gardasil. I then provide the argument that gendering Gardasil as a "women's technology" is done intentionally by Merck, which has a history of making vi profits a priority over responsibly treating patient health. I conclude by providing detailed suggestions on how Merck can augment their current ad series to de-gender Gardasil to become more medically responsible, and break out of the cycle of portraying men and women using damaging and outdated stereotypes. Instead, my suggestions for changes to Gardasil's advertising approach would make the vaccine's messages appeal to all audiences at risk.

Retrocyclin RC-101, a θ-defensin with lectin-like properties, potently inhibits infection by many HIV-1 subtypes by binding to the heptad repeat (HR)-2 region of gp41 and preventing six-helix bundle formation. In the present study, we used in silico computational exploration to identify residues of HR2 that interacted with RC-101 and then analyzed the HIV-1 Sequence Database at LANL for residue variations in the HR1 and HR2 segments that could plausibly impart in vivo resistance. Docking RC-101 to gp41 peptides in silico confirmed its strong preference for HR2 over HR1, and implicated residues crucial for its ability to bind HR2. We mutagenized these residues in pseudotyped HIV-1 JR.FL reporter viruses, and subjected them to single round replication assays in the presence of 1.25-10ug/ml RC-101. Except for one mutant that was partially resistant to RC-101, the other pseudotyped viruses with single-site cationic mutations in HR2 manifested absent or impaired infectivity or retained wild-type susceptibility to RC-101. Overall, these data suggest that most mutations capable of rendering HIV-1 resistant to RC-101 will also exert deleterious effects on the ability of HIV-1 to initiate infections - an interesting and novel property for a potential topical microbicide.

Although nucleic acid tests (NATs) for human immunodeficiency virus (HIV) exhibit many advantages, such as early detection and viral load quantification, over immunological assays, their widespread use is limited by their demand for high-level infrastructure, sophisticated equipment, and advanced staff competence. Furthermore, when quantifying viral loads of patients, it has been reported that these assays can underestimate viral quantities by 22- to 100-fold due to primer-template mismatches in more divergent HIV subtypes. Therefore, we have developed a cost-effective and sequence-independent assay for the detection and quantification of HIV utilizing a modified nucleic acid sequence-based amplification (NASBA) protocol coupled to an electrochemical DNA biosensor. The modified NASBA reaction involves the addition of a 22-nucleotide tag between the T7 RNA Polymerase Promoter and the hybridizing region of the reverse primer. As a result, this tag will be placed at the 5' end of each amplicon regardless of the target sequence. We then designed the DNA sensor to hybridize to this segment of the amplicon specifically. Therefore, hybridization, and subsequently, detection, is dependent only on the presence of this tag and not the viral RNA sequence itself. As a result, the issue of underestimating viral loads is eliminated as multiple primers can be used in one reaction without having to use multiple sensors. The use of an isothermal NASBA technique and a reusable gold-disc electrode for the sensor helps drive down the cost of the assay by eliminating the need for thermocyclers and fluorometers used by conventional NATs.

Autoimmune and thyroid conditions account for a substantial proportion of the morbidity and mortality experienced in the United States, affecting >40 million Americans combined. Co-occurrence of both an autoimmune and thyroid condition is also likely, particularly among women. Epidemiologic studies on both sets of conditions have examined many risk factors, including demographic, lifestyle, genetic, and environmental risk factors. However, one area which has been neglected is the effect of early life exposures on the development of autoimmune and thyroid conditions. To investigate the potential association between an individual’s birth weight (by category;in uteroand early childhood, extending the hypothesis to include thyroid conditions.

Obesity is recognized as an important risk factor for the development of many chronic diseases such as hypertension, Type 2 diabetes mellitus (T2DM) cardiovascular disease, cancer, renal disease, neurologic dysfunction, metabolic syndrome and asthma (3, 4). Circulating serum adiponectin levels in obese asthmatics have been reported to be low. Therefore, we aimed to investigate the role of adiponectin in a mucus hypersecretion model and hypothesized that adiponectin would decrease IL-13 induced MUC5AC expression from differentiated NHBE cells and that increasing concentrations of IL-13 would cause a decrease in Adipo R1 and Adipo R2 expression. MUC5AC expression with exposure to adiponectin was not significant. However, mRNA expression of Adipo R1 and Adipo R2 was significantly decreased by stimulation of IL-13 for acute (24 hours) and chronic (14 days) exposure. Therefore, the obese state and specifically IL-13 concentration could play a role in Adipo R1 and Adipo R2 expression within NHBE cells.