Browsing by Subject "HIV"
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Item A Review of the Literature on Faith-Based Organization's HIV/AIDs Care and Prevention Efforts in Sub-Saharan Africa(2005-05-01) Rojas, Zeida G.; Lurie, Sue; Urrutia-Rojas, XimenaRojas, Zeida G., A Review of the Literature on Faith-Based Organization’s HIV/AIDS Care and Prevention in Sub-Saharan Africa. Master of Public Health (Community Health), May 2005, 83 pp., 20 diagrams, bibliography, 10 titles. The thesis assesses the role of faith-based organizations (FBOs) involved in HIV/AIDS related care and prevention activities in Sub-Saharan Africa. Do FBOs have the ability to address the multi-faceted syndrome that HIV/AIDS brings to an individual, their family and community? Can FBOs be effective partners to carry out prevention and care initiatives? Faith-based organizations are generally overlooked as potential partners and leaders in the fight against HIV/AIDS. FBOs are often the only genuine nongovernmental organizations in many rural parts of poor countries, or at a minimum, they are the strongest and most influential. Due to their organizational networks, FBOs are able to mobilize people and resources, and to reach rural or isolated areas. Evidence of current FBO efforts demonstrates that FBOs have the ability to address the multifaceted effects of HIV/AIDS and can become indispensable partners for government health agencies and NGOs.Item A Social Vaccine for HIV/AIDS: Investing in Our Future Through Curriculum-Based Education in Tanzania: A Conceptual Implementation Model(2009-05-01) Tenende, Tunu; Lykens, KristineAn estimated 10 million young people worldwide are living with HIV with a yearly incidence of 2 million in the 15-24 year old population. This is nearly half of all new cases that are being reported. Worldwide roughly 39.5 million people are dying of Acquired Immune Deficiency Syndrome (AIDS) or are infected with the Human Immuno-deficiency Virus (HIV). Of these 39.5 million people, 24.7 million live in Africa; a strong majority in resource scarce countries. In Tanzania alone there are 1.4 million people living with HIV making the percentage of people with HIV at 6.2%. With other prevention efforts that have been underway in the past few years, we have thus far been able to see a drop in people affected from 7.0% to what is now current at 6.2%. With an understanding that the country is capable of making stride in lowering incidence which will ultimately lower prevalence down the line, focus on the countries youth has been an avenue to explore. Though there are studies that have looked into prevention methods and even more specifically have looked into curriculum-based prevention programs, there have not been any that focus on a comprehensive approach that includes seeking governmental support and mandating a change in the curriculum to focus on HIV/AIDS education. Based on literature review of components that can influence the development, implementation and incorporation of an HIV/AIDS prevention program in Tanzania, this paper concludes that a curriculum-based HIV/AIDS prevention program is the most effective and efficient manner to reach the community and bring about the biggest change.Item Astrocyte Elevated Gene-1, a Novel Modulator of Astrocyte Function: Implications for neuroAIDS, aging and glioblastoma(2013-12-01) Vartak, Neha; Ghorpade, AnujaVartak-Sharma, Neha N., Astrocyte elevated gene-1, a novel modulator of astrocyte function: Implications for NeuroAIDS, aging and glioblastoma. Doctor of Philosophy (Biomedical Sciences), Nov, 2013, 180 pp., 1 table, 40 illustrations, 336 bibliographies. Recent attempts to analyze human immunodeficiency virus (HIV)-1-induced gene expression changes in astrocyte identified a multifunctional oncogene, astrocyte elevated gene-1 (AEG-1), as an HIV-1 and tumor necrosis factor-inducible transcript. Subsequently, due to its homology to mouse breast cancer metastasis protein, metadherin, AEG-1 was largely implicated in carcinogenesis of diverse cancer types. However, the role of AEG-1 in astrocytes, the original cell type in which AEG-1 was first identified, still remains to be investigated. In the present study, we identified AEG-1 as a novel modulator of astrocyte function during reactive astrogliosis, neuroinflammation and neurodegeneration, and elucidated its implications in NeuroAIDS, aging and cancer. Our in vitro and in vivo studies recognized AEG-1 modulation of astrocyte migration and proliferation towards the wound site, thereby regulating astrocyte wound healing, a fundamental homeostatic function of astrocytes. Further, AEG-1 expression analyses in HIV-1+ and HIV-1 encephalitic human brain tissues provided the necessary physiological evidence for AEG-1 induction upon HIV-1 neuroinvasion. Herein, we identified AEG-1 as an inflammatory response gene and as an important upstream regulator of NF-κB signaling in astrocytes. Our results demonstrated AEG-1 cytoplasmic and nuclear interaction with NF-κB p65 subunit, which was crucial for NF-κB nuclear translocation, thereby regulating astrocyte neuroinflammation. In the same study, we also identified AEG-1 as a novel regulator of astrocyte glutamate clearance, an important determinant of neurocognitive CNS function, by modulating the expression of the key glutamate transporter, excitatory amino acid transporter 2. Analyses of AEG-1 expression in the cognitive centers of the brain of aging individuals demonstrated AEG-1 age-dependent expression in the human brain, which further proposed a role for AEG-1 in cellular oxidative stress responses. Herein, we identified a novel antioxidant cytoprotective role of AEG-1 in astrocytes and astrocytoma cells. Cellular localization studies by confocal microscopy revealed AEG-1 localization to the dense fibrillar components of the nucleolus in response to injury or oxidative stress, suggesting AEG-1 implication in ribosomal RNA processing. Our results demonstrated AEG-1 regulation of catalase activation and Nrf2 stabilization in response to oxidative stress and further elucidated AEG-1 modulation of Nrf2 nuclear translocation, the first step in antioxidant cellular defense mechanisms. The results presented in this thesis provide insight into the role of oncogene AEG-1 in human astrocytes and ameliorates our understanding of astrocyte-mediated processes in normal and disease-relevant pathologies, ranging from HIV-1-associated neurocognitive disorders and traumatic CNS injuries to primary neoplasms of the brain.Item Astrocyte TIMP-1: Regulation and Gene Delivery in HAND(2018-12) Joshi, Chaitanya R.; Ghorpade, Anuja; Clark, Abbot F.; He, Johnny J.; Sumien, Nathalie; Rickards, Caroline A.Despite antiretroviral therapy, HIV-associated neurocognitive disorders (HAND) persist in 30-70% of patients. During HAND, elevated matrix metalloproteinases (MMPs) in the brain exacerbate the disease by blood-brain barrier breakdown, neuroinflammation and direct neurotoxicity. Tissue inhibitors of metalloproteinases (TIMPs) counter MMP activity. In the brain, TIMP-1 is primarily produced by astrocytes in response to injury or inflammation. However, TIMP-1 is downregulated during chronic inflammation in astrocytes and in HIV encephalitis brain tissues. We propose that restoring astrocyte TIMP-1 levels could mitigate neurodegeneration due to its MMP-inhibitory and -independent neuroprotective functions. HIV-1 non-productively infects astrocytes, which express viral proteins such as transactivator of transcription (Tat). As Tat mimics aspects of HAND by direct and indirect mechanisms, glial fibrillary acidic protein (GFAP) promoter-restricted Tat expressing (GT-Tg) mice were used to model HAND in our studies. Prolonged astrocyte Tat expression in GT-Tg mice resulted in HAND-relevant behavioral impairments characterized by higher anxiety, lower ambulation, impaired spatial learning, and memory. Importantly, behavioral deficits were accompanied by altered brain MMP/TIMP balance. Our data from GT-Tg mouse model confirmed neurocognitive decline and TIMP-1 dysregulation in the context of HAND. As TIMP-1 was downregulated with prolonged Tat expression in mice, we focused on replenishing TIMP-1 via gene delivery to the brain using cationic polymers. Polyethylenimine (PEI) is a highly efficient polymer for transfecting mammalian cells, however, high cytotoxicity restricts its use. Hence, PEI was modified using arginine (A) and stabilized with polyethylene glycol (P) to produce multiple AnPn analogues. AnPn analogues were biocompatible and successfully delivered reporter genes to primary neural cells. Select AnPn led to sustained reporter gene expression in human astrocytes and in mouse brains. In order to restrict gene expression to astrocytes, truncated GFAP promoters were used to drive gene expression. Subsequently, GFAP promoters were modified enhancing their activity and increasing gene expression. Lastly, successful polymer-mediated GFAP promoter-driven TIMP-1 gene delivery was demonstrated in human astrocytes. Overall, these findings enhance our understanding HIV-1 Tat-mediated TIMP-1 regulation, provide a novel therapeutic TIMP-1 gene delivery system, and pave the way for future investigations geared towards preclinical translation of TIMP-1-based HAND therapy.Item CHEMOKINE CXCL8 MODULATES HIV-1 REPLICATION IN HUMAN MONOCYTE-DERIVED MACROPHAGES(2013-04-12) Mamik, Manmeet K.Purpose: Chemokine CXCL8 is an important neutrophil chemoattractant implicated in various neurodegenerative disorders. We previously reported that HIV-1 infection is linked to upregulation of CXCL8 in brain tissue. We also reported that SHP2 and MAPK pathways regulate the expression of CXCL8 in human astrocytes. In the post-ART era, low level of productive replication of HIV-1 in brain is a critical component of neuropathogenesis regulation. The present study investigated the effect of CXCL8 on productive infection of HIV-1 in human monocytes-derived macrophages (MDM). Methods: Human MDM were infected with blood and brain-derived HIV-1 isolates, HIVADA and HIVJR-FL. HIV p24 levels were assayed from culture supernatants with ELISA. DNA was isolated from infected cells treated with/without CXCL8 (10-100ng/ml) and amplified for two-LTR circles, as a measure of integration We employed human promonocytic cell line U937 as an efficient transfection system. U937 cells were transfected with pHIV-LTR and promoter activity was measured by luciferase reporter assay. Results: Treatment with CXCL8 led to significant upregulation (p<0.01) in HIV-1 p24 levels in supernatants of HIV-infected MDM, as determined by ELISA. Reverse transcriptase (RT) activity was significantly increased (p<0.01) in HIV-infected MDM treated with CXCL8. We compared the formation of 2-LTR circles in CXCL8 treated vs untreated HIV-infected MDM by RT-PCR, as a measure of viral genome integration. Transient transfection of U937 cells with HIV-LTR construct containing luciferase reporter gene resulted in increased luciferase activity when treated with CXCL8. Conclusions: The results show that CXCL8 mediates productive infection of HIV-1 in MDM and induces HIV-1 LTR promoter activity in U937 cells. Detailed understanding of the mechanisms involved could aid in therapeutic intervention strategies by modulating levels of this chemokine in the brain.Item CROFELEMER FOR HIV-ASSOCIATED DIARRHEA: SUSTAINED EFFICACY, SAFETY, AND ADHERENCE DURING A 6-MONTH RANDOMIZED, PLACEBO-CONTROLLED TRIAL(2014-03) Clay, Patrick G.Combination antiretroviral therapy (cART) in HIV+ individuals has increased life expectancy; however, increased ART exposure is also associated with adverse effects that can negatively impact overall health, work productivity, and healthcare resource utilization. Diarrhea, an adverse event (AE) of ART, remains a substantial burden associated with reduced quality of life and may result in ART nonadherence or treatment failure. Reported prevalence of diarrhea in the community is up to 28% of patients receiving cART. Crofelemer, extracted from the stem bark latex of the Croton lechleri tree, is a minimally absorbed, first-in-class antidiarrheal agent indicated for the symptomatic relief of noninfectious diarrhea in adults with HIV/AIDS receiving ART. Crofelemer inhibits chloride ion (Cl–) secretion and accompanying high-volume water loss in secretory diarrhea via dual inhibition of cystic fibrosis transmembrane conductance regulator and calcium-activated Cl– channels in the intestinal epithelium. Purpose (a): To assess patient adherence to and efficacy and safety of crofelemer 125 mg twice daily for up to 6 months. Methods (b): Randomized, phase 3, double-blind, placebo-controlled, 2-stage trial (Figure 1). The optimal crofelemer (Fulyzaq™, Salix Pharmaceuticals, Inc., Raleigh, NC, USA) dose (125, 250, or 500 mg twice daily) was determined in stage 1. Based on the stage 1 interim efficacy and safety analysis, crofelemer 125 mg twice daily was selected as the optimal dose and was evaluated in additional patients in stage 2; data for crofelemer 125 mg twice daily were combined (stage 1 and 2). Primary efficacy measure: percentage of patients achieving clinical response, defined as ≤2 watery stools per week for ≥2 of 4 weeks during the placebo-controlled phase (1-sided analysis). Results (c): Demographic and baseline characteristics were similar between crofelemer 125 mg twice daily (n = 136) and placebo (n = 138) groups (Table 1); patients had a mean of 2.7 to 3.0 watery stools per day (ie, >18 watery stools per week). Primary measure: A significantly larger percentage of patients treated with crofelemer achieved clinical response compared with placebo (17.6% vs 8.0%; P < 0.01). Continued and sustained improvement in weekly clinical response and a mean improvement from baseline in diarrhea symptoms was observed in patients who continued to receive crofelemer 125 mg twice daily during the placebo-free phase. Conclusions (d): Crofelemer 125 mg twice daily was efficacious and well tolerated, and improvements in noninfectious diarrhea symptoms appeared to be durable for at least 6 months in an HIV+ population receiving stable cART. Treatment of diarrhea in HIV+ individuals may provide several important benefits, such as improvement in cART adherence.Item Doctors, Patients, and Adherence to HIV Medication: Findings of the Communication, Communities, and Health Study(2008-05-01) Seater, Margaret; Kimberly Fulda; Kathryn CardarelliThis study is about whether doctors have the potential to influence adherence by forming a solid patient-doctor relationship. This study is also about health disparities; specifically, if racialized life experiences have any association with either adherence or the formation of a solid patient-doctor relationship. Self-reported racial discrimination was shown to be a risk factor for non-adherence (OR 4.725, p-value [less than] 0.05), while compassionate behavior on the part of the clinician predicted adherence (OR 0.062, p-value [less than] 0.1 trend). Future directions include applying for extramural funding to conduct a clinical trial emphasizing communication as a way to eliminate health disparities. In the long term, the goal of medical educators should be to recruit more non-white physicians in order to further eliminate health disparities.Item Effect of 2018 American College of Cardiology/American Heart Association Guideline Change on Statin Prescription for People Living with HIV(Elsevier B.V., 2023-03-28) Pan, Meng; Agovi, Afiba Manza-A.; Anikpo, Ifedioranma O.; Fasanmi, Esther O.; Thompson, Erika L.; Reeves, Jaquetta M.; Thompson, Caitlin T.; Johnson, Marc E.; Golub, Vitaly; Ojha, Rohit P.The American College of Cardiology (ACC)/American Heart Association (AHA) guidelines were updated in 2018 to explicitly recommend statin use for primary cardiovascular disease prevention among people living with HIV (PLWH), but little is known about the effect of this guideline change. We aimed to assess the effect of the 2018 ACC/AHA guideline change on statin prescription among PLWH. We used data from an institutional HIV registry to identify PLWH aged 40-75 years, engaged in HIV care between June 2016 and May 2021, had a LDL cholesterol between 70 and 189 mg/dl, 10-year atherosclerotic cardiovascular disease (ASCVD) risk score >/=7.5%, no prior statin prescription, and no history of diabetes or ASCVD. Our outcome of interest was a new statin prescription within 12 months of eligibility. We estimated standardized risk difference (RD) with 95% confidence limits (CL) by comparing prescription probabilities before and after guideline change. Our study population comprised 251 PLWH (171 before, 80 after the guideline change), of whom 57% were aged <55 years, 82% were male, and 45% were non-Hispanic black. The standardized 12-month statin prescription risk was 43% (95% CL: 31%, 60%) after the guideline change and 19% (95% CL: 13%, 26%) before the guideline change (RD = 25%, 95% CL: 9.1%, 40%). Our results suggest that the 2018 ACC/AHA guideline change increased statin prescription among PLWH, but a sizable proportion of eligible PLWH were not prescribed statin. Future studies are needed to identify strategies to enhance implementation of statin prescription guidelines among PLWH.Item Evaluation of NK Cell – Astrocyte Interactions: Potential Role in HIV-Associated Neurocognitive Disorders and HIV- Associated Dementia(2015-05-01) Bowen, Kelly E.; Mathew, Porunelloor A.; Mathew, Stephen O.; Hodge, Lisa M.NK cells play important roles in immunity against pathogens and cancer. NK cell functions are regulated by inhibitory and activating receptors binding corresponding ligands on the surface of target cells. During pathological conditions, NK cells were shown to be recruited to the CNS and could impact CNS physiology by killing glial cells and by secreting IFN-g. Astrocytes are intimately involved in immunological and inflammatory events occurring in the CNS and reactive astrogliosis is a key feature in HIV-associated neurocognitive disorders (HAND). There is little data on NK cell-astrocyte interactions and ligands expressed on astrocytes that could impact NK cell function. This study aimed to identify NK-associated ligands expressed by human astrocytes that confer this NK-directed cytotoxicity of astrocytes and assay the cytotoxicity differences in presence and absence of HIV 3S peptide. Using a fusion protein consisting of the extracellular domain of NKp44 fused to Fc portion of human IgG, we determined the expression of a novel ligand for NKp44 (NKp44L) on astrocytes. Incubation of astrocytes with 3S peptide downregulated NKp44L expression on astrocytes implicating protection from NK mediated killing. Thus, our study demonstrated that NKp44 has a protective effect on astrocytes from NK cell mediated killing during HIV infection. Astrocytes could also secrete cytokines that affect the expression of NK receptors on NK cells. We evaluated the expression of receptors on NK cells after co-culture with astrocytes. CD38 expression was increased on primary NK cells after incubation with astrocytes. CD38 is expressed on both NK cells and astrocytes and has an important implication in HIV-1 infection. Blocking CD38 signaling in our studies decreased astrocyte lysis, suggesting CD38 signaling has important implications in NK-astrocyte interactions. Future studies providing novel insights into the role of NK cells in the pathogenesis of HAND and other brain disorders might result in the development of NK cell based therapies for brain pathologies.Item Functional Role of 2B4+ CD8+ T Cells Against HIV Epitopes(2009-07-01) Aldy, Kim N; Mathew, StephenAcquired immune deficiency syndrome (AIDS), caused by Human Immunodeficiency Virus (HIV) is the most devastating global health problem. Long Term Non Progressors (LTNP) are seropositive individuals that have delayed progression to AIDS. Potent anti-HIV CD8+ T cell responses are associated with LTNP’s; therefore, an effective cytotoxic T cell (CTL) response is vital for the immune system to control HIV. Human leukocyte antigen (HLA) B14 and HLA B27 types are commonly present in LTNP’s. 2B4 (CD244) is expressed on NK cells, monocytes, basophils, eosinophils, and a small population of CD8+ T cells. Although the expression of 2B4+CD8+ T cells has been shown to increase during HIV disease progression, the role of 2B4+ CD8+ T cells has not been established. Studying the differences in activation and cytotoxic activity of 2B4+ and 2B4¯ CD8+ T cells will expand our knowledge of this receptor in T cells, which has not been fully addressed in the past. HIV infected patients could benefit from immunotherapy efforts using the knowledge we gain from 2B4 studies in T cells. In addition, the information gathered from this study can be used for in vivo studies of this receptor in natural HIV infection. The immunomodulatory effects of Corticotrophin releasing hormone and Epinephrine on 2B4- and 2B4+ CD8+ T cells during activation and the effects on cytotoxicity have not been published to date. Knowledge about their effects will allow us to learn the function of these hormones that are released due to stress during immune regulation in HIV disease.Item HIV AND HCV INFECTION OF THE BRAIN IN HUMANIZED MICE(2013-04-12) Zhang, ZiugenPurpose: In this study, we wished to determine the possibility of HIV and HCV infections in the brains of these mice in preparation of using the mice to study HIV and HCV co-infection of the brain and the roles of the co-infection in HIV/HCV-associated neurological diseases . Methods: Human CD34+ human hematopoietic stem cells (HSC) and hepatocyte progenitors were co-transplanted into the Balb/CRag2-/-𝛄C-null mice, which led to efficient engraftment of human leukocytes and hepatocytes. These humanized mice were infected with HCV alone or HCV and HIV. Two months after infection, livers and brains of these mice were collected. Half of the brain was fixed and paraffin-embedded for immunostaining for HIV p24 and HCV core; the other half of the brain was used for DNA and RNA extraction. Polymerase Chain Reaction (PCR) was performed to detect HIV DNA; Strand-specific Reverse Transcription Polymerase Chain Reaction (RT-PCR) was performed to detect positive-strand and negative strand HCV RNA. Results: Three of four mice in HCV mono-infected group, one of five mouse in the HCV/HIV co-infected group showed severe fibrosis in liver. The HCV core protein and HIV P24 protein were detected in the brain, although faint. All 5 mouse infected with HIV were detected for HIV DNA in the brain. All 4 HCV mono-infected mice and all 5 HCV/HIV co-infected mice were detected for positive-strand HCV RNA in their brains, while only 3 of those brains were detected for negative-strand HCV RNA. Conclusions: These results showed that HCV and HIV are both capable of gaining access into the brain and replicating in the brain of these mice and suggest that humanized mice could be used to study the effects of HCV infection and HCV/HIV co-infection on the brain dysfunction.Item HIV AND NUTRITION IN A COMMUNITY SETTING(2014-03) Leber, Julie; DeHaven, MarkPurpose (a): A balanced diet and good nutrition help maintain a strong immune system for resisting disease and contribute to improved quality of life. Weight loss, wasting, and malnutrition are common problems which can contribute to HIV disease progression. With recent advances in effective antiretroviral medications, good nutrition can help those infected with HIV to better process their many medications. Diet (and exercise) may help control other symptoms such as diarrhea, nausea, and fatigue, and other metabolic abnormalities such as high blood sugar, cholesterol, and triglycerides. The purpose of this project was to assess the behaviors, knowledge, and attitudes related to modifiable lifestyle factors for improving health outcomes among residents of an HIV/AIDS living facility in Fort Worth (Samaritan House). This study will provide the baseline for understanding the potential value in making future nutritional interventions within the living facility. Methods (b): The Samaritan House in Fort Worth is dedicated to creating a supportive community providing housing and resources for positive change in the lives of persons living with HIV/AIDS and other special needs. A conglomerate of validated questionnaires was administered to Samaritan House residents in order to assess their knowledge, attitudes, and behaviors with regards to nutrition, physical activity, depression, and smoking. Results (c): Results showed that residents intake patterns did not meet the dietary recommendations with regards to fat, fruits, vegetables, and fiber, and that the majority of residents worry considerably about their health but do not change their eating habits because of it. The majority of residents answered that motivations for healthy behavior were driven by internal rather than external factors. 64% screened positive for depression. 54% were active smokers, but 41% had tried to quit in the past year. With regards to physical activity, 33% of residents had a high level, 33% had a medium level, and 33% had a low level. Conclusions (d): Nutritional and lifestyle renovation are a potential source of improvement at Samaritan House. This results of this study will be used to provide a foundation by which later studies can be conducted that examine the effects of dietary interventions (through education and influence of the charitable donations providing food) on the holistic health of this population, which will serve to improve the quality of life and prevention of disease.Item HIV Related Risk Behaviors: A Comparitive Study of Urban, Suburban, and Rural U.S. Adolescents(2006-08-01) Patil, Godavari D.; Karan Singh; Sejong Bae; Francise Soto MasGodavari D. Patil, HIV Related Risk Behaviors: A Comparative Study of Urban, Suburban, and Rural U.S. Adolescents. Masters of Public Health (Biostatistics), August 2006, 120 pp., 29 tables, References, 209 titles. This explorative study YRBS 2003 data provides the prevalence of HIV-related risky sexual behaviors and predictors of such behaviors across gender, race/ethnicity, and metro status (N=15, 214) during 2003. Overall, more urban male adolescents engaged in health-compromising behaviors. A significant association was found between gender, race/ethnicity, and metro status and sexual behaviors and associated risk behaviors such as alcohol, drug use, and mental health indicators. These associated risk behaviors were not only associated among themselves and with sexual behaviors variables but also turned out to be responsible predictor variables for HIV related sexual risk behaviors. Minority groups especially black adolescents were at higher risk of contracting HIV infection as having multiple sexual partners was highest (8 fold) among black adolescents compared to mixed & other race, and Hispanic adolescents. Suburban adolescents were nearly two times more likely that rural and urban adolescents to having multiple partners. Results indicated that younger the age more the involvement in sexual and other risky behaviors.Item IDENTIFYING HIGH RESPONDER POPULATIONS TO CROFELEMER FOR TREATMENT OF NONINFECTIOUS DIARRHEA IN HIV+ INDIVIDUALS(2014-03) Clay, Patrick G.Background: Diarrhea remains a significant burden in some HIV+ individuals. Crofelemer is a first-in-class, minimally absorbed, botanically derived drug indicated for symptomatic relief of noninfectious diarrhea in adults with HIV/AIDS taking antiretroviral therapy. This analysis examined the efficacy of crofelemer 125 mg BID in specific subpopulations. Methods: In a 4-week, phase 3, randomized, double-blind, placebo-controlled trial in HIV+ adults receiving antiretroviral therapy, crofelemer 125 mg BID (n = 136) was compared with placebo (n = 138) for reducing noninfectious diarrhea. The primary efficacy endpoint (≤2 watery stools/wk for ≥2 of 4 weeks) was examined in subgroups that differed according to demographic (sex, age, race, time since HIV diagnosis, and duration of diarrhea) and baseline characteristics (severity of diarrhea, prior antidiarrheal medication [ADM] use, stool consistency, viral load, CD4+ cell count, and use of protease inhibitors). Results: At baseline, patients experienced (range of means) 18.9-21.0 watery stools/week, and investigators attributed diarrhea most commonly (75%) to antiretroviral therapy (ritonavir-containing agents, 34%-36%; efavirenz/tenofovir/emtricitabine, 15%-22%) or HIV enteropathy (24%). Across all subgroups considered, treatment effects favored crofelemer. Pronounced treatment differences (crofelemer – placebo) were observed for patients with prior use of ≥2 ADMs (+28%; P 2 watery stools/day (+10%; P = 0.03), diarrhea duration [greater than] 2 years (+11%; P = 0.03), and use of protease inhibitors (+11%; P = 0.021). Conclusions: These findings suggest that the crofelemer antidiarrheal effect is robust and generalizable across patient subpopulations. Purpose (a): To determine which patients with HIV/AIDS are most likely to respond to treatment with crofelemer 125 mg twice daily. Methods (b): Patients and Study Design: HIV+ adults taking a stable ART regimen for ≥4 weeks, with a history of diarrhea (persistently loose stools despite regular antidiarrheal medication use or ≥1 watery bowel movement per day without regular antidiarrheal medication use for ≥1 month). Exclusion criteria included CD4+ cell count/μL and positive gastrointestinal biopsy, culture, or stool test for infectious agents in the previous 4 months. Randomized, double-blind, phase 3 trial of crofelemer 125 mg or placebo administered twice daily for 4 weeks. Assessments: Primary efficacy measure: percentage of patients achieving clinical response, defined as ≤2 watery stools per week for ≥2 of 4 weeks of treatment. Assessment of efficacy was based on patient diaries, which recorded symptoms of diarrhea (eg, stool consistency, stool frequency), adherence to study drug and ART, and use of antidiarrheal and prohibited medications; results were entered daily using an interactive voice response system. Stool consistency score was computed using the mean of all reported stool scores for 1 day; stools were scored using a scale ranging from 1 to 5 (1 = very hard; 5 = watery). Statistical Analyses: Efficacy was assessed in the intention-to-treat population composed of all randomized patients receiving ≥1 dose of study drug (1-sided for primary efficacy analysis, based on technique described by Posch et al13; 2-sided for subgroup analyses). Fisher’s exact test was used for comparisons between crofelemer and placebo for subgroup analyzed. Subgroup analyses were not corrected for multiple comparisons. Results (c): Patient Population: Demographic and baseline characteristics were similar between the 2 groups. Patients in the crofelemer and placebo treatment arms were of similar age (mean, 45 vs 44.8 y), sex (male, 84.6% vs 84.1%), and race/ethnicity (white, 39.0% vs 42.0%; black, 37.5% vs 38.4%; Hispanic, 22.8% vs 18.1%; American Indian/Alaskan Native, 0.7% vs 0%; other, 0% vs 1.5%). Patients in both groups averaged >18.9 watery bowel movements per week. Historical use of antidiarrheal medication was common, reported by 79 patients (58%) receiving crofelemer 125 mg and 83 patients (60%) receiving placebo. Loperamide-containing agents were historically used by 53 (39%) and 60 (43%) patients treated with crofelemer and placebo, respectively. Although not permitted during the 4 weeks of treatment, concomitant use of antidiarrheal medications (eg, loperamide, diphenoxylate /atropine, bismuth subsalicylate) were reported by 1.1% and 4.0% of patients receiving crofelemer or placebo, respectively. Efficacy: A significantly greater percentage of patients achieved clinical response during treatment with crofelemer 125 mg twice daily versus placebo (P = 0.0096; Figure 2). Conclusions (d): Subgroup analyses confirm crofelemer 125 mg twice daily provides robust clinical response across multiple patient subpopulations. Crofelemer was particularly effective in patients with factors associated with severe diarrhea, consistent with its antisecretory effects on intestinal chloride channels.Item Intercellular Nef transfer and HIV-1 infection of astrocytes(2015-05-01) Luo, Xiaoyu; He, Johnny J.; Ghorpade, Anuja; Wordinger, Robert J.Acquired immune deficiency syndrome (AIDS) is a pandemic caused by human immunodeficiency virus type 1 (HIV-1). It is a major health issue in many parts of the world ever since its discovery in 1981. The most devastating effect of HIV-1 infection is the graduate loss of CD4+ T cells, which eventually leads to the dysfunction of the immune system, susceptibility to opportunistic infections and cancer. HIV-1 Nef protein is long known as an essential pathogenic factor for HIV-1/AIDS pathogenesis. A few recent studies including ours have demonstrated that Nef can be transferred to neighboring cells and alters the function of these cells. However, the underlying mechanism of intercellular Nef transfer is in dispute. In the first part of our study, we characterized two potential underlying mechanisms for intercellular Nef transfer: direct cell-cell contact and exosomes using several complementary strategies and a panel of exosomal markers. First, we showed that Nef was transferred from Nef-expressing or HIV-infected CD4+ T lymphocytes to CD4+ T lymphocytes and astrocytes, and that the transfer was mainly associated with tunneling nanotube formation. Then we determined that Nef enhanced virological synapse formation and induced cytoskeleton re-arrangement and cell surface protrusions, suggesting that Nef promotes the establishment of intercellular connection and communication between infected cells and uninfected cells. Thirdly, we examined the possibility of Nef transfer through exosomes. In the exosome uptake assay, Nef transfer was undetectable while exosome marker CD81 transferred rapidly. In contrast, Nef was detected in crude exosomes collected from Nef-transfected 293T. In addition, two different populations of exosomes were successfully separated by OptiPrep gradient fractionation and determined as AChE+/CD81low/TSG101low exosomes and AChE- /CD81high/TSG101high exosomes. We determined that Nef was selectively secreted into the AChE+/CD81low/TSG101low population. Lastly, microscopic imaging showed no significant Nef detection in exosomal vesicle-like structures in and out the cell. Taken together, this study shows that Nef transfer requires direct cell-cell contact such as tunneling nanotubes, not cell-free exosomes. In addition, this study reveals existence of two types of exosomes: AChE+/CD81low/TSG101low exosomes and AChE/CD81high/TSG101high exosomes. In the second part, we characterized HIV-1 infection of astrocytes. Astrocytes are the most abundant cells in the central nervous system (CNS) and play important roles in HIV-1/neuroAIDS. Detection of HIV-1 proviral DNA, RNA and early gene products but not late structural gene products in astrocytes in vivo and in vitro indicates that astrocytes are susceptible to HIV-1 infection albeit in a restricted manner. We, as well as others have shown that cell-free HIV-1 is capable of entering CD4- astrocytes through human mannose receptor-mediated endocytosis. In this study, we took advantage of several newly developed fluorescence protein-based HIV-1 reporter viruses and further characterized HIV-1 interaction with astrocytes. First, we found that HIV-1 was successfully transferred to astrocytes from HIV-infected CD4+ T cells in a cell-cell contact- and gp120-dependent manner. In addition, we demonstrated that compared to endocytosis-mediated cell-free HIV-1 entry and subsequent degradation of endocytosed virions, cell-cell contact between astrocytes and HIV-infected CD4+ T cells led to robust HIV-1 infection of astrocytes but retained the restricted nature of viral gene expression. Furthermore, we showed that HIV-1 latency was established in astrocytes. Lastly, we demonstrated that infectious progeny HIV-1 was readily recovered from latently infected astrocytes in a cell-cell contact-mediated manner. Taken together, our studies point to the importance of the cell-cell contact-mediated HIV-1 interaction with astrocytes and provide direct evidence to support the notion that astrocytes are HIV-1 latent reservoirs in the CNS.Item INTERCELLULAR TRANSFER OF HIV-1 NEF AMONG CD4 T CELLS IS MEDIATED BY DIRECT CELL-CELL CONTACT(2013-04-12) Luo, XiaoyuPurpose: HIV-1 Nef is an essential pathogenic factor for acquired immunodeficiency syndrome disease progression. It binds to over 30 putative cellular components and initiates a great variety of functions. Recent studies have demonstrated that Nef can be transferred from infected cells to uninfected cells and leads to significant changes on the bystander cells. This intercellular Nef transfer is possibly linked to the massive depletion of CD4 T cells in disease progression. However, the underlying mechanism for the intercellular Nef transfer is still not fully understood. In this study, by using advanced tools, we tested two potential mechanisms for intercellular Nef transfer among CD4 T cells including direct Nef transfer through cell-cell contact and Nef secretion in the form of exosomes. Methods: Nef-GFP Jurkat cells were co-cultured with red dye-labeled uninfected Jurkat cells for 8 hours. Cell mixture was analyzed under florescence microscope. To determine the percentage of Nef transfer during HIV infection, HIV and HIV Nef-deleted virus-infected jurkat cells were collected when cells reached 50% infected. The percentage of Nef+ cells in uninfected population and how it affected by cell density was analyzed by Immunostaining and flow-cytometry. In order to test if exosomes are involved in intercellular Nef transfer among CD4 T cells, we utilized optiprep gradient centrifugation to separate exosomes from viral particles and then detect Nef expression in exosome fractions. Results: Nef was detected in the bystander Jurkat cells after co-cultured with Nef-expressing /infected Jurkat cells. Nef was transferred through nanotubes and filopodia bridges by forming connections between infected cells and uninfected cells. There was about 3-10% of uninfected cells that were found to be Nef+ in infected Jurkat cells (50% infection). The intercellular Nef transfer was enhanced by increasing the cell density. Using optiprep gradient centrifugation, virus was successfully separated with exosomes and Nef was only detected in the virus fractions but not exosome fractions. Conclusions: Taken together, our studies showed that Nef can be transferred to uninfected bystander CD4+ T cells either from Nef-expressing cells or from HIV-infected cells. This intercellular Nef transfer is mediated by direct cell-cell contact possibly through synapse or nanotube like structures. Exosome is not involved in the intercellular Nef transfer.Item POPULATION PHARMACOKINETIC ANALYSIS DEMONSTRATES NO DRUG-DRUG INTERACTIONS BETWEEN CROFELEMER, A NOVEL TREATMENT FOR NONINFECTIOUS DIARRHEA IN HIV+ INDIVIDUALS, AND ANTIRETROVIRAL THERAPY(2014-03) Clay, Patrick G.Crofelemer, a minimally absorbed, botanically derived, first-in-class antidiarrheal agent, was approved by the US Food and Drug Administration in December 2012 at a dose of 125 mg twice daily for symptomatic relief of noninfectious diarrhea in adult patients with HIV/AIDS on ART. Secretory diarrhea, particularly noninfectious diarrhea, is associated with combination antiretroviral therapy (ART) therapy. It is characterized by increased secretion of chloride ions and the movement of sodium and water into the intestinal lumen as a result of cyclic adenosine monophosphate (cAMP)–stimulated cystic fibrosis transmembrane conductance regulator (CFTR) and calcium-activated chloride channel (CaCC) activation by HIV, ART, or bacterial enterotoxins. Crofelemer is a dual inhibitor of CFTR and CaCC in the intestinal epithelium. For patients with HIV/AIDS receiving combination ART, comorbidities requiring management or treatment with other therapies increase the potential for drug-drug interactions (DDI) with ART6; clinically significant DDI have been reported in 27% to 40% of patients with HIV/AIDS taking ART. Significant DDI with ART may be caused by induction or inhibition of major metabolic pathways, or by drug transporters, resulting in reductions in ART efficacy, safety, and tolerability. Purpose (a): To determine the potential for crofelemer to alter the pharmacokinetics (PK) of concomitantly administered ART agents in HIV+ patients in the ADVENT trial. Methods (b): HIV+ adults taking a stable ART regimen for ≥4 weeks, with a history of diarrhea (ie, persistently loose stools despite regular antidiarrheal medication use or ≥1 watery bowel movement per day without regular antidiarrheal use for ≥1 month) and CD4+ count ≥100 cells/μL. ADVENT is a randomized, double-blind, two-stage phase 3 trial (Figure 1) conducted from October 2007 to January 2011 Pharmacokinetic Assessments. A multiple-trough sampling design14 was used to assess population PK. Sample collection for PK assessments occurred at baseline, prior to dosing at randomization, at the end of the placebo-controlled phase (ie, week 4), and at the end of the study (ie, week 24). Immune status was evaluated by analysis of CD4+ cell count and HIV viral load. Crofelemer concentrations were assayed by validated high-performance liquid chromatography with fluorescence detection (Celerion, Lincoln, NE, USA); ART concentrations were assayed by Tandem Labs (Salt Lake City, UT, USA) and the University of North Carolina Clinical Pharmacology and Analytical Chemistry Core Facility (Chapel Hill, NC, USA). Results (c): More than 97.5% of patients in the PK population (n = 353) had received ≥ 3 combination ART regimens before the study. Patients received 126 different combinations of ART; the most frequently used combination ART was EFV, FTC, and TNF (n = 60). All 6 of the most common ARTs demonstrated marked exposure variability during the study (Figure 2). Week 4 and 24 ART steady-state trough drug concentrations in patients receiving crofelemer, regardless of crofelemer dose, were comparable with concentrations obtained during the crofelemer-free period (for example, TNF in Figure 3). Comparable results were demonstrated with RTV, FTC, 3TC, LPV, and EFV (Table 2). Crofelemer had no statistically significant effect on the PK of the 6 most commonly used ARTs in ADVENT, as assessed by the Bonferroni correction approach (Table 2). Administration of crofelemer had no negative impact on clinical immune parameters (HIV viral load and CD4+ cell counts) • In >96% of patients, crofelemer concentrations were below the limit of quantitation (50 ng/mL). Conclusions (d): Crofelemer had no significant effect on the PK of the most frequently-used ART evaluated in this study. Consistent with the absence of effects on ART PK, crofelemer did not adversely affect ART efficacy, based on HIV viral load or CD4+ cell counts. Crofelemer was not systemically absorbed to a significant extent.Item Qualitative Analysis on HIV/AIDS Clinical Trial Recruitment Marketing Practices(2022-05) Dapoz, Anthony M., Jr.; Simecka, Jerry W.Recruitment continues to be one of the largest barriers to clinical trial success. Based on review of clinical trial marketing literature, there are no marketing industry standards established for recruitment of HIV/AIDS clinical trials. The purpose of this study was to begin finding optimal marketing recruitment strategies in HIV/AIDS clinical trials research. A literature review yielded 20 articles detailing recruitment marketing practices. A ClinicalTrials.gov search found 57 trials meeting inclusion criteria. Data extracted included recruitment strategies, original enrollment goals, estimated enrollment goals and actual enrollment goals. Regression analysis found no significant relationship between marketing strategies and recruitment rate. Added financial and managerial considerations should be incorporated with a robust site level marketing campaign to optimize recruitment potential.Recruitment continues to be one of the largest barriers to clinical trial success. Based on review of clinical trial marketing literature, there are no marketing industry standards established for recruitment of HIV/AIDS clinical trials. The purpose of this study was to begin finding optimal marketing recruitment strategies in HIV/AIDS clinical trials research. A literature review yielded 20 articles detailing recruitment marketing practices. A ClinicalTrials.gov search found 57 trials meeting inclusion criteria. Data extracted included recruitment strategies, original enrollment goals, estimated enrollment goals and actual enrollment goals. Regression analysis found no significant relationship between marketing strategies and recruitment rate. Added financial and managerial considerations should be incorporated with a robust site level marketing campaign to optimize recruitment potential.Item SAFETY AND TOLERABILITY OF CROFELEMER 125 MG TWICE DAILY IN THE TREATMENT OF NONINFECTIOUS DIARRHEA IN HIV-SEROPOSITIVE PATIENTS ON ANTIRETROVIRAL THERAPY: RESULTS FROM A PHASE 3, 48-WEEK OPEN-LABEL STUDY(2014-03) Clay, Patrick G.Diarrhea remains a substantial health concern in patients with HIV in the era of combination antiretroviral therapy (ART), despite the decline in opportunistic infectious diarrhea. For example, noninfectious diarrhea attributable to ART-related adverse events (AEs) has a reported prevalence in the community of up to 28% of patients with HIV who are receiving ART. Diarrhea in patients with HIV is commonly a leaky-flux or secretory diarrhea, which is characterized by increased secretion of chloride ions (Cl-) and subsequent sodium and water flow into the gastrointestinal lumen. Crofelemer (Fulyzaq™, Salix Pharmaceuticals, Inc., Raleigh, NC, USA) is a minimally absorbed, first-in-class, botanically derived drug indicated for the symptomatic relief of noninfectious diarrhea in adults with HIV. Following oral dosing of crofelemer, plasma concentrations of the drug were below the level of quantification in [greater than] 99% of patients with HIV and diarrhea. Crofelemer is a dual inhibitor of the cyclic adenosine monophosphate (cAMP)-stimulated cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel and the calcium-activated Cl- channel (CACC). In a phase 3, double-blind, placebo-controlled trial (ADVENT), crofelemer 125 mg twice daily significantly reduced diarrhea in patients with HIV receiving ART compared with placebo, and the safety profile was comparable to placebo for up to 24 weeks. Purpose (a): To evaluate the long-term (up to 48 weeks) safety and tolerability of crofelemer 125 mg twice daily for the treatment of noninfectious diarrhea in patients with HIV. Methods (b): Phase 3, multicenter, open-label study of crofelemer 125 mg twice daily for up to 48 weeks. Results (c): Overall, 189 patients (75.6%) experienced ≥1 AE during this open-label study; the majority of AEs (90.5% of patients) were mild or moderate in intensity. The most commonly reported AEs were infection-related (eg, upper respiratory tract [16.8%], intestinal parasitic [12.4%], Giardia [8.0%]; none drug-related) or gastrointestinal-related (eg, nausea [5.6%], constipation [5.6%]) –Most patients (10 of 14 [71.4%]) reporting a constipation AE also used ADMs during the study. Only 9 (3.6%) patients reported diarrhea as an AE •AEs considered at least possibly related to study drug occurred in 9.2% of patients; the most commonly reported were constipation (3.6%), abdominal distension (2.0%), abdominal pain (1.2%), and flatulence (1.2%). No deaths occurred during the study; serious AEs occurred in 20 patients (8.0%), including infection in 10 patients; none were considered drug-related. Conclusions (d): Crofelemer 125 mg twice daily was well tolerated with a low incidence of AEs in HIV-seropositive patients with noninfectious diarrhea receiving ART, which is consistent with the minimal absorption of crofelemer. In this study, minimal clinical deterioration of immune status was observed for up to 48 weeks, suggestive of adherence to ART regimens and continued ART efficacy.Item Structural Determinants of Health Among Transgender Populations: Policing, Sex Work, and HIV(2023-12) Webb, Nathaniel J.; Kline, Nolan; Griner, Stacey; Rossheim, Matthew; Yockey, AndrewTransgender (trans) populations experience disproportionately high rates of HIV infection compared to cisgender populations. Additionally, due to overlapping layers of discrimination in education, housing, healthcare, and employment, trans populations are more likely to engage in a criminalized form of work, such as sex work. Policing has been identified as a potential structural determinant of HIV infection among individuals engaging in sex work. Trans populations, including those engaging in sex work, are more likely to interact with police and experience some form of police violence. This dissertation investigated policing as a potential structural determinant of HIV status and HIV testing among trans individuals who engage in sex work. Unadjusted and adjusted logistic regression analyses were utilized to identify associations between HIV status/HIV testing and police interactions. Mediation analyses were utilized to investigate police interactions as a potential mediating variable between HIV status/HIV testing and trans individuals who have engaged in street-based sex work. Statistically significant associations were identified between police interactions and HIV status in unadjusted (OR: 2.564, 95% CI: 1.166, 5.641, p-value = 0.019) and adjusted (aOR: 12.055, 95% CI: 3.076, 47.232, p-value <0.001) logistic regression models. Additionally, police interactions were not identified as a statistically significant mediating variable between HIV status/HIV testing and trans individuals engaging in street-based sex work. These findings suggest policing may act as a contributing factor towards HIV infection among trans individuals engaged in sex work, but further research is needed to elucidate this interaction. HIV infection prevention interventions need to include an intersectional lens that incorporates trans identities and address the structural issues that trans populations experience including discrimination in housing, employment, and healthcare.