Comparison of In-Person Versus Telephone Interviews for Early Syphilis and HIV Partner Services in King County, Washington (2010-2014)

Thesis (Master's)--University of Washington, 2016-08

megaTAL-mediated Gene Editing at the CCR5 locus

Thesis (Ph.D.)--University of Washington, 2016-08

Identification of protein targets of nevirapine reactive metabolites using click chemistry and mass spectrometry-based differential proteomics

Abstract : Adverse drug reactions (ADRs) are undesirable effects caused after administration of a single dose or prolonged administration of drug or result from the combination of two or more drugs. Idiosyncratic drug reaction (IDR) is an adverse reaction that does not occur in most patients treated with a drug and does not involve the therapeutic effect of the drug. IDRs are unpredictable and often life-threatening. Idiosyncratic reaction is dependent on drug chemical characteristics or individual immunological response. IDRs are a major problem for drug development because they are usually not detected during clinical trials. In this study we focused on IDRs of Nevirapine (NVP), which is a non-nucleoside reverse transcriptase inhibitor used for the treatment of Human Immunodeficiency Virus (HIV) infections. The use of NVP is limited by a relatively high incidence of skin rash. NVP also causes a rash in female Brown Norway (BN) rats, which we use as animal model for this study. Our hypothesis is that idiosyncratic skin reactions associated with NVP treatment are due to post-translational modifications of proteins (e.g., glutathionylation) detectable by MS. The main objective of this study was to identify the proteins that are targeted by a reactive metabolite of Nevirapine in the skin. The specific objectives derived from the general objective were as follow: 1) To implement the click chemistry approach to detect proteins modified by a reactive NVP-Alkyne (NVP-ALK) metabolite. The purpose of using NVP-ALK was to couple it with Biotin using cycloaddition Click Chemistry reaction. 2) To detect protein modification using Western blotting and Mass Spectrometry techniques, which is important to understand the mechanism of NVP induced toxicity. 3) To identify the proteins using MASCOT search engine for protein identification, by comparing obtained spectrum from Mass Spectrometry with theoretical spectrum to find a matching peptide sequence. 4) To test if the drug or drug metabolites can cause harmful effects, as the induction of oxidative stress in cells (via protein glutathionylation). Oxidative stress causes cell damage that mediates signals, which likely induces the immune response. The results showed that Nevirapine is metabolized to a reactive metabolite, which causes protein modification. The extracted protein from the treated BN rats matched 10% of keratin, which implies that keratin was the protein targeted by the NVP-ALK.

Att leva med HIV/AIDS i Norden

Bakgrund: Humant immunbristvirus, HIV, är ett virus som angriper kroppens immunsystem. Acquired immunodeficiency syndrome, AIDS, utvecklas från HIV efter lång tid utan behandling. I samhället råder det okunskap och rädsla kring sjukdomen vilket skapar stigmatisering och diskriminering som dagligen påverkar de som lever med HIV/AIDS. Attityden mot HIV-smittade personer är hos många negativ. Det finns sjuksköterskor som inte vill behandla HIV-smittade på grund av rädsla och okunskap. Att få diagnosen är livsomställande och att handskas med det är inte lätt. Syfte: Syftet var att beskriva hur det är att leva med HIV/AIDS i Norden. Metod: Litteraturöversikten baserades på sex kvantitativa och sju kvalitativa artiklar för att få en fördjupad kunskap och en överblick över kunskapsläget för hur det är att leva med HIV/AIDS i Norden. Resultat: I resultatet framkommer det tre kategorier som tillsammans ger en överblick om hur det är att leva med HIV/AIDS. Dessa kategorier är; anpassning till ett nytt liv, hålla hemligt eller komma ut och begränsad sexualitet. Stigmatisering påverkade personernas syn på sig själva och deras beslut kring avslöjandet av sjukdomen. Slutsats: Personer som lever med HIV bemöts ofta med okunskap, fördomar och avståndstagande, därför är bemötandet avgörande. Litteraturöversikten bidrar med information och kunskap som senare kan användas av vårdpersonal för att få en djupare förståelse för personer som lever med HIV. Sjuksköterskan bör ha kunskap om HIV/AIDS för att på bästa sätt kunna ge god omvårdnad, då okunnighet och fördomar i samhället ansågs vara värre än själva sjukdomen.Nyckelord:

Health and mood among HIV‐positive outpatients attending an ART Clinic of a University Hospital

To evaluate how individuals at different stages of infection with HIV perceive their health status and its association with mood states. With the introduction of Highly Active Antiretroviral Therapy in 1996, the quality of life of people living with HIV has improved. However, the literature emphasises the negative effects of the disease on the mental health of individuals suffering from this condition and the high incidence of depression among infected individuals. Although people diagnosed and living with HIV are overwhelmed by emotions, we found that various emotional manifestations are understudied within this group of patients. A cross-sectional study was conducted in an outpatient unit of a University Hospital (antiretroviral therapy clinic), with a consecutive sample composed of 152 patients. Data were collected through a questionnaire used to assess the sociodemographic and clinical characteristics, the Short Form (36) Health Survey, and the Profile of Mood States scale. The health status negatively affects the role at the emotional and mental health dimensions. The participants showing a worse health condition than in the previous year had higher levels of tension/anxiety, depression/dejection, fatigue/inertia and confusion/bewilderment. The stage of disease and the profile of mood state emerged as independent phenomena. The results of this study indicate that nurses worldwide should be aware of the emotional aspects (negative emotions strongly impact health) related to the subjective perception of a worsening health status, regardless of the stage of the disease.

Um caso pediátrico de tuberculose miliar: achados raros e evolução atípica

Introdução: A tuberculose miliar resulta da disseminação linfohematogénica do Mycobacterium tuberculosis, sendo uma manifestação grave da infeção. Caso clínico: Criança de 9 anos, género feminino, com his¬tória de febre prolongada. O diagnóstico de tuberculose miliar foi colocado após telerradiografia torácica com infiltrado reticu¬lonodular difuso bilateral, e corroborado pelo achado de tubér¬culos coroideus no olho direito e visualização de bacilos álcool¬ ¬ácido resistentes em amostra de suco gástrico. Detetaram¬-se tuberculomas cerebrais na ressonância magnética. Isolou¬se Mycobacterium tuberculosis multissensível em amostras de suco gástrico. Após mais de 40 dias de tratamento, persistia a febre e baciloscopia positiva. Foi excluída infeção pelo vírus da imunodeficiência humana. Não foram detetadas complicações. Posteriormente, a evolução clínica foi favorável. Discussão/Conclusão: A tuberculose mantém-¬se um diag¬nóstico relevante na criança com febre prolongada. A associa¬ção da imagem torácica, baciloscopias positivas e tubérculos coroideus foram fundamentais para a celeridade do diagnóstico e implementação do tratamento. Reforça¬-se a importância de manter elevado índice de suspeição para uma patologia que tem tratamento.

Paradoxical Deterioration During Anti-Tuberculous Therapy in Non-HIV-Infected Patients with Pleural Tuberculosis: A Pragmatic Approach

We report a case of paradoxical deterioration. A male patient diagnosed with pleural tuberculosis, but who was not infected with human immunodeficiency virus (HIV), experienced clinical deterioration 3 weeks after the initiation of anti-tuberculous treatment. After other diagnoses were ruled out, a paradoxical response to treatment was established and the patient was started on systemic corticosteroids. Paradoxical response to treatment should be considered in patients with clinical deterioration after they start on anti-tuberculous treatment.

HIV Reverse Transcriptase Fidelity And Inhibition Are Modulated By Divalent Cations In A Concentration-Dependent Manner In Vitro

Human immunodeficiency virus (HIV) rapidly evolves through generation and selection of mutants that can escape drug therapy. This process is fueled, in part, by the presumably highly error prone polymerase reverse transcriptase (RT). Fidelity of polymerases can be influenced by cation co-factors. Physiologically, magnesium (Mg2+) is used as a co-factor by RT to perform catalysis, however, alternative cations including manganese (Mn2+), cobalt (Co2+), and zinc (Zn2+) can also be used. I demonstrate here that fidelity and inhibition of HIV RT can be influenced differently, in vitro, by divalent cations depending on their concentration. The reported mutation frequency for purified HIV RT in vitro is typically in the 10-4 range (per nucleotide addition), making the enzyme several-fold less accurate than most polymerases. Paradoxically, results examining HIV replication in cells indicate an error frequency that is ~10 times lower than the error rate obtained in the test tube. Here, I reconcile, at least in part, these discrepancies by showing that HIV RT fidelity in vitro is in the same range as cellular results, in physiological concentrations of free Mg2+ (~0.25 mM). At low Mg2+, mutation rates were 5-10 times lower compared to high Mg2+ conditions (5-10 mM). Alternative divalent cations also have a concentration-dependent effect on RT fidelity. Presumed promutagenic cations Mn2+ and Co2+ decreases the fidelity of RT only at elevated concentrations, and Zn2+, when present in low concentration, increases the fidelity of HIV-1 RT by ~2.5 fold compared to Mg2+. HIV-1 and HIV-2 RT inhibition by nucleoside (NRTIs) and non-nucleoside RT inhibitors (NNRTIs) in vitro is also affected by the Mg2+ concentration. NRTIs lacking 3'-OH group inhibited both enzymes less efficiently in low Mg2+ than in high Mg2+; whereas inhibition by the “translocation defective RT inhibitor”, which retains the 3ʹ-OH, was unaffected by Mg2+ concentration, suggesting that NRTIs with a 3ʹ-OH group may be more potent than other NRTIs. In contrast, NNRTIs were more effective in low vs. high Mg2+ conditions. Overall, the studies presented reveal strategies for designing novel RT inhibitors and strongly emphasize the need for studying HIV RT and RT inhibitors in physiologically relevant low Mg2+ conditions.

Prevalence and associated factors of TB/HIV co-infection among HIV Infected patients in Amhara region, Ethiopia.

Background: Tuberculosis is one of the world’s most common causes of death in the era of Human immunodeficiency virus. The purpose of this study was to determine the prevalence and associated factors of TB/HIV co-infection. Methods: Hospital based retrospective studies were conducted among adult HIV-positive patients. Logistic regression method and Chi square test were applied. Results: A total of 571 HIV positive study participants were enrolled. Of these, 158 (27.7%) were found to have pulmonary tuberculosis. Lower baseline CD4 count<200cell/μl, patients who drunk alcohol, patients who were ambulatory at the initiation of ART, patients whose marital status was single were significant predictors for increased risk of tuberculosis in PLWHIV (P <0.05). Non smoker patients, patients in WHO clinical stage I, patients in WHO clinical stage II and ownership of the house had significant protective benefit against risk of TB (P <0.05). Conclusion: The prevalence of TB/HIV co-infection in adults on ART in our study was moderately high. Having advanced clinical status and presence of risk factors were found to be the predicting factors for co-infection. The health office should open TB/HIV co-infection units in the hospitals and health workers should be cautious when a patient has an advanced disease.

Cerebro-meningeal infections in HIV-infected patients: a study of 116 cases in Libreville, Gabon.

Background: Cerebro-meningeal pathology is common in human immunodeficiency virus (HIV) infection and the aetiology is often difficult to ascertain with certainty. Objective: To describe the major suspected and identified causes of meningeal or encephalitic syndromes in HIV infection in Libreville, Gabon. Methods: A descriptive study using clinical records of patients hospitalised in the Department of Medicine in the Military Hospital of Libreville (Gabon) between January 2006 and May 2010. Clinical features were evaluated using multivariable logistic regression to evaluate association with the outcome of a clinical improvement or death. Results: The most frequent neurological symptoms were reduced level of consciousness (54.3%), headache (55.2%), motor deficit (38.7%), and convulsions (36.2%). Cerebral toxoplasmosis represented 64.7% of diagnoses, followed by cryptococcal neuromeningitis in 12.9% of cases. Tuberculoma was diagnosed in 4 cases and lymphoma in 2 cases. In 9.5% of cases, no aetiology was determined. Toxoplasmosis treatment led to clinical improvement in 69.3% of cases with suspected cerebral toxoplasmosis. Overall mortality was 39.7%. Conclusion: The diagnosis of neurological conditions in HIV positive patients is difficult, particularly in a low-resource setting. A trial of treatment for toxoplasmosis should be initiated first line with all signs of neurological pathology in a patient infected with HIV.

Five years retrospective cohort analysis of treatment outcomes of TB-HIV patients at a PEPFAR/DOTS Centre in South Eastern Nigeria.

Background: Human immunodeficiency virus (HIV) associated tuberculosis (TB) remains a major global public health challenge, with an estimated 1.4 million patients worldwide. Co-infection with HIV leads to challenges in the diagnosis and treatment of patients. Objectives: The aim of this study was to assess treatment outcomes of a cohort of smear positive TB-HIV co-infected patients over a five-year study period. Methods: A retrospective cohort study of 600 smear-positive tuberculosis patients registered at the chest unit of the University of Nigeria Teaching Hospital, Enugu from January 2008 to December 2012 was done. The data was analyzed using SPSS Version 17. Results: One hundred and three (17.2%) of the patients were co-infected with TB/HIV, while 398 (66.3%) and 99 (16.5%) were HIV negative and unknown respectively. Among the co-infected patients, 45(43.7%) were cured as against 222(55.8%) in the TBHIV negatives (Z=4.53, p=0.000, 95%CI= 0.12-0.34). Respectively in the TB-HIV co-infected and TB-HIV negative patients, treatment completed were 21(20.4%) and 71(17.8%) (Z=9.15, p=0.000, 95%= 0.4035-0.60); defaulted 19(18.5%) vs 70 (17.6%) (Z=9.29, p=0.000, 95%CI=0.42-0.60), died 10(9.7%) vs. 6(1.5%) (Z=1.22, p=0.224, 95%CI= -0.0286-0.1086), and failures were 1(0.9%) vs. 7(1.8%) (Z=2.48, p=0.013, 95%CI=0.04-0.10). Treatment success rate was lower in TB-HIV co-infected patients, 64.1% compared to TB-HIV negative patients with 73.6%. Also those that defaulted among the TB-HIV co-infected patients (18.5%) were higher than 17.6% among TB-HIV negative patients, a difference of 0.9%. Conclusion: Findings demonstrate that HIV co-infection affects TB treatment outcomes adversely. Treatment adherence, timely and sustained access to antiretroviral therapy for TB/HIV co-infected patients are important.

Profile and treatment outcomes of patients with tuberculosis in Northeastern Ethiopia: a cross sectional study.

Background: Data on the epidemiology of tuberculosis and its treatment outcomes were incomplete in the study area and this study was done to fill this gap. Methods: Institution based cross sectional study was conducted from January 2011 to December 2014. A total of 949 TB patients who were on treatment in North Eastern Ethiopia, Eastern Amhara region were included. Data was analyzed using SPSS version 20. Frequency, percentages and means were used to present data. To assess the associations of treatment outcomes with sex, age, type of TB and human immunodeficiency virus (HIV), logistic regression was used. Results: The proportion of smear positive and negative pulmonary TB, and extra pulmonary TB were 187/949 (19.7%), 322/949 (33.9%) and 440/949 (46.4%), respectively. Treatment success rate was 853/949 (89.9%). Smear positive pulmonary TB and TB/HIV co-infections were significantly associated with unsuccessful treatment outcome, P≤ 0.002. Conclusion: Extra pulmonary TB was the most prevalent types of TB followed by smear negative pulmonary TB. Treatment success rate was above the WHO target of 85%. The causes for the high proportion of smear negative PTB and EPTB should be further investigated. Special emphasis should be put on smear positive PTB patients and TB/HIV co-infected patients to decrease unsuccessful treatment outcome and TB transmissions.

Transient hyperglycemia in patients with tuberculosis in Tanzania: implications for diabetes screening algorithms.

BACKGROUND: Diabetes mellitus (DM) increases tuberculosis risk while tuberculosis, as an infectious disease, leads to hyperglycemia. We compared hyperglycemia screening strategies in controls and patients with tuberculosis in Dar es Salaam, Tanzania. METHODS: Consecutive adults with tuberculosis and sex- and age-matched volunteers were included in a case-control study between July 2012 and June 2014. All underwent DM screening tests (fasting capillary glucose [FCG] level, 2-hour CG [2-hCG] level, and glycated hemoglobin A1c [HbA1c] level) at enrollment, and cases were tested again after receipt of tuberculosis treatment. Association of tuberculosis and its outcome with hyperglycemia was assessed using logistic regression analysis adjusted for sex, age, body mass index, human immunodeficiency virus infection status, and socioeconomic status. Patients with tuberculosis and newly diagnosed DM were not treated for hyperglycemia. RESULTS: At enrollment, DM prevalence was significantly higher among patients with tuberculosis (n = 539; FCG level > 7 mmol/L, 4.5% of patients, 2-hCG level > 11 mmol/L, 6.8%; and HbA1c level > 6.5%, 9.3%), compared with controls (n = 496; 1.2%, 3.1%, and 2.2%, respectively). The association between hyperglycemia and tuberculosis disappeared after tuberculosis treatment (adjusted odds ratio [aOR] for the FCG level: 9.6 [95% confidence interval {CI}, 3.7-24.7] at enrollment vs 2.4 [95% CI, .7-8.7] at follow-up; aOR for the 2-hCG level: 6.6 [95% CI, 4.0-11.1] vs 1.6 [95% CI, .8-2.9]; and aOR for the HbA1c level, 4.2 [95% CI, 2.9-6.0] vs 1.4 [95% CI, .9-2.0]). Hyperglycemia, based on the FCG level, at enrollment was associated with tuberculosis treatment failure or death (aOR, 3.3; 95% CI, 1.2-9.3). CONCLUSIONS: Transient hyperglycemia is frequent during tuberculosis, and DM needs confirmation after tuberculosis treatment. Performance of DM screening at tuberculosis diagnosis gives the opportunity to detect patients at risk of adverse outcome.

Review: Head and neck squamous cell carcinoma in sub-Saharan Africa

Aim Review the literature from 1990 to 2013 to determine known anatomic sites, risk factors, treatments, and outcomes of head and neck squamous cell carcinoma (HNSCC) in sub-Saharan Africa. Methods Using a systematic search strategy, literature pertaining to HNSCC in sub-Saharan Africa was reviewed and patient demographics, anatomic sites, histology, stage, treatment, and outcomes were abstracted. The contributions of human immunodeficiency virus (HIV), human papillomavirus (HPV) and behavioural risk factors to HNSCC in the region were assessed. Results Of the 342 papers identified, 46 were utilized for review, including 8611 patients. In sub-Saharan Africa, the oropharyngeal/oral cavity was found to be the most common site, with 7750 cases (90% of all cases). Few papers distinguished oropharyngeal from oral cavity, making identification of possible HPV-associated oropharyngeal squamous cell carcinoma (SCC) difficult. SCC of the nasopharynx, nasal cavity, or paranasal sinuses was identified in 410 patients (4.8% of all cases). Laryngeal SCC was found in 385 patients (4.5% of all cases), and only 66 patients (0.8% of all cases) with hypopharyngeal SCC were identified. In 862 patients with data available, 43% used tobacco and 42% used alcohol, and reported use varied widely and was more common in laryngeal SCC than that of the oropharyngeal/oral cavity. Toombak and kola nut use was reported to be higher in patients with HNSCC. Several papers reported HIV-positive patients with HNSCC, but it was not possible to determine HNSCC prevalence in HIV-positive compared to negative patients. Reports of treatment and outcomes were rare. Conclusions The oropharyngeal/oral cavity was by far the most commonly reported site of HNSCC reported in sub-Saharan Africa. The roles of risk factors in HNSCC incidence in sub-Saharan Africa were difficult to delineate from the available studies, but a majority of patients did not use tobacco and alcohol.

Exploring fertility decisions among pregnant HIVpositive women on antiretroviral therapy at a health centre in Balaka, Malawi: A descriptive qualitative

Background The proportions of women of reproductive age living with the human immunodeficiency virus (HIV) vary between different regions of the world, with significantly higher proportions in sub-Saharan Africa. Family planning is one of the major issues that couples and families affected with HIV must confront. We aimed to assess the cultural and social factors associated with childbearing and family planning knowledge, decisionmaking, and practices among HIV-positive pregnant women attending antenatal clinic at a health centre in Balaka, Malawi. Methods This was a qualitative descriptive study carried out at Kalembo Health Centre in Balaka. A purposive sampling technique was used to select pregnant women enroled in the antiretroviral therapy (ART) programme. A sample size of thirty-five women was decided upon after data saturation. Qualitative inquiry was used during data collection. Data were analysed using systematic text condensation, while numbers and percentages were generated using Microsoft Excel. Results Out of 35 participants, 20 were aged between 25 and 34 years, and 18 had been married at least three times. All 35 women wished to have their own biological child. Factors, reported by participants, that promote childbearing included: the desire to please their husbands, fear of losing their husbands to others if they did not bear children, the knowledge that ART would help prevent their children from acquiring the virus, the desire to prove to others that they can also bear children, and a lack of family planning leading to unplanned pregnancies. Conclusions The factors that lead to pregnancies among women on ART in Balaka ranged from assured safety of the child from HIV, lack of contraception, to other factors related to their partners. The authors recognize and support the freedom for women to become pregnant and bear children, and, in the context of HIV infection, fertility and reproductive services should include a comprehensive approach towards addressing issues of HIV and AIDS and childbearing among infected women.