Reducing misclassification bias in cervical dysplasia risk factor analysis with p16-based diagnoses.

OBJECTIVE: Conventional hematoxylin and eosin (HE)-based diagnoses have been the reference standard for cervical cancer risk factor analyses. However, this HE-based method is known to have modest interobserver reproducibility and only moderate predictive value. In contrast, more recent immunohistochemical-based diagnoses using the neoplastic marker p16 are known to improve diagnostic accuracy. Our objective was to test whether p16-based diagnoses would significantly affect high-grade dysplasia (cervical intraepithelial neoplasia 2+) risk factor analysis compared with the current reference standard (HE). MATERIALS AND METHODS: Retrospective cohort of 500 index cases were randomly selected from a series of more than 5,000 cervical biopsies performed at Kaiser Permanente Northwest from 1997 to 2003 after a patient's first abnormal cervical Pap smear (positive for atypical squamous cells of undetermined significance). Subjects were subsequently excluded if they did not have at least 5 years of clinical follow-up, including cervical biopsies, or 3 reproducibly negative Pap smears. This yielded 358 cases for risk factor analysis. The index biopsies and all follow-up biopsies were immunostained for p16 and the proliferation marker Ki-67, which were then independently reviewed by 2 pathologists blinded to clinical outcomes. Data were analyzed by χ test and logistic regression modeling. RESULTS: We observed clinically significant diagnostic errors in 22% of index biopsies. Improved accuracy using p16 strengthened the risk estimate of low family income for cervical intraepithelial neoplasia 2+ (odds ratio = 1.71, 95% confidence interval = 1.09-2.63) compared with HE-based diagnoses (odds ratio = 1.12, 95% confidence interval = 0.72-1.72). The addition of Ki-67 staining did not significantly influence these results. CONCLUSIONS: p16-based diagnoses may affect the power of risk factor analysis, especially when using small cohorts.

Patient activation and improved outcomes in HIV-infected patients.

BACKGROUND: The Patient Activation Measure (PAM) assesses several important concepts in chronic care management, including self-efficacy for positive health behaviors. In HIV-infected populations, better self-efficacy for medication management is associated with improved adherence to antiretroviral medications (ARVs), which is critically important for controlling symptoms and slowing disease progression. OBJECTIVE: To determine 1) characteristics associated with patient activation and 2) associations between patient activation and outcomes in HIV-infected patients. DESIGN: Cross-sectional survey. PARTICIPANTS: 433 patients receiving care in four HIV clinics. METHODS: An interviewer conducted face-to-face interviews with patients following their HIV clinic visit. Survey data were supplemented with medical record abstraction to obtain most recent CD4 counts, HIV viral load and antiretroviral medications. MAIN MEASURES: Patient activation was measured using the 13-item PAM (possible range 0-100). Outcomes included CD4 cell count > 200 cells/mL(3), HIV-1 RNA < 400 copies/mL (viral suppression), and patient-reported adherence. KEY RESULTS: Overall, patient activation was high (mean PAM = 72.3 [SD 16.5, range 34.7-100]). Activation was lower among those without vs. with a high school degree (68.0 vs. 74.0, p < .001), and greater depression (77.6 lowest, 70.2 middle, 68.1 highest tertile, p < .001). There was no association between patient activation and age, race, gender, problematic alcohol use, illicit drug use, or social status. In multivariable models, every 5-point increase in PAM was associated with greater odds of CD4 count > 200 cells/mL(3) (aOR 1.10 [95 % CI 1.01, 1.21]), adherence (aOR 1.18 [95 % CI 1.09, 1.29]) and viral suppression (aOR 1.08 [95 % CI 1.00, 1.17]). The association between PAM and viral suppression was mediated through adherence. CONCLUSIONS: Higher patient activation was associated with more favorable HIV outcomes. Interventions to improve patient activation should be developed and tested for their ability to improve HIV outcomes.

CD8+ T cells provide an immunologic signature of tuberculosis in young children.

RATIONALE: The immunologic events surrounding primary Mycobacterium tuberculosis infection and development of tuberculosis remain controversial. Young children who develop tuberculosis do so quickly after first exposure, thus permitting study of immune response to primary infection and disease. We hypothesized that M. tuberculosis-specific CD8(+) T cells are generated in response to high bacillary loads occurring during tuberculosis. OBJECTIVES: To determine if M. tuberculosis-specific T cells are generated among healthy children exposed to M. tuberculosis and children with tuberculosis. METHODS: Enzyme-linked immunosorbent spot assays were used to measure IFN-γ production in response to M. tuberculosis-specific proteins ESAT-6/CFP-10 by peripheral blood mononuclear cells and CD8(+) T cells isolated from Ugandan children hospitalized with tuberculosis (n = 96) or healthy tuberculosis contacts (n = 62). MEASUREMENTS AND MAIN RESULTS: The proportion of positive CD8(+) T-cell assays and magnitude of CD8(+) T-cell responses were significantly greater among young (<5 yr) tuberculosis cases compared with young contacts (P = 0.02, Fisher exact test, P = 0.01, Wilcoxon rank-sum, respectively). M. tuberculosis-specific T-cell responses measured in peripheral blood mononuclear cells were equivalent between groups. CONCLUSIONS: Among young children, M. tuberculosis-specific CD8(+) T cells develop in response to high bacillary loads, as occurs during tuberculosis, and are unlikely to be found after M. tuberculosis exposure. T-cell responses measured in peripheral blood mononuclear cells are generated after M. tuberculosis exposure alone, and thus cannot distinguish exposure from disease. In young children, IFN-γ-producing M. tuberculosis-specific CD8(+) T cells provide an immunologic signature of primary M. tuberculosis infection resulting in disease.

An Assessment of Serological Assays for SARS-CoV-2 as Surrogates for Authentic Virus Neutralization.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in late 2019 and has since caused a global pandemic resulting in millions of cases and deaths. Diagnostic tools and serological assays are critical for controlling the outbreak, especially assays designed to quantitate neutralizing antibody levels, considered the best correlate of protection. As vaccines become increasingly available, it is important to identify reliable methods for measuring neutralizing antibody responses that correlate with authentic virus neutralization but can be performed outside biosafety level 3 (BSL3) laboratories. While many neutralizing assays using pseudotyped virus have been developed, there have been few studies comparing the different assays to each other as surrogates for authentic virus neutralization. Here, we characterized three enzyme-linked immunosorbent assays (ELISAs) and three pseudotyped vesicular stomatitis virus (VSV) neutralization assays and assessed their concordance with authentic virus neutralization. The most accurate assays for predicting authentic virus neutralization were luciferase- and secreted embryonic alkaline phosphatase (SEAP)-expressing pseudotyped virus neutralizations, followed by green fluorescent protein (GFP)-expressing pseudotyped virus neutralization, and then the ELISAs. IMPORTANCE The ongoing COVID-19 pandemic is caused by infection with severe acute respiratory syndrome virus 2 (SARS-CoV-2). Prior infection or vaccination can be detected by the presence of antibodies in the blood. Antibodies in the blood are also considered to be protective against future infections from the same virus. The "gold standard" assay for detecting protective antibodies against SARS-CoV-2 is neutralization of authentic SARS-CoV-2 virus. However, this assay can only be performed under highly restrictive biocontainment conditions. We therefore characterized six antibody-detecting assays for their correlation with authentic virus neutralization. The significance of our research is in outlining the advantages and disadvantages of the different assays and identifying the optimal surrogate assay for authentic virus neutralization. This will allow for more accurate assessments of protective immunity against SARS-CoV-2 following infection and vaccination.

Effect of mode of delivery on the incidence of urinary incontinence in primiparous women.

OBJECTIVE: To estimate the effect of mode of delivery on the incidence of urinary incontinence in primiparous women. METHODS: A population-based survey was mailed to all Oregon women who delivered a liveborn neonate in a 1-year period. Data were collected on urinary incontinence, childbirth experience, and other risk factors for incontinence at 3-6 months postpartum. Univariable analyses were conducted using t tests and Wilcoxon rank-sum tests for continuous variables and chi tests for categorical variables. Logistic regression analyses were used to estimate odds ratios and 95% confidence intervals for demographic and clinical risk factors. RESULTS: A total of 15,787 women completed the survey, for a response rate of 39%. Of these women, 5,599 were primiparous, completed the survey in the desired timeframe, submitted information on their urinary continence, and did not have incontinence before pregnancy. A total of 955 (17.1%) reported leakage of urine. Women who had vaginal deliveries were more likely to have urinary incontinence than women who had cesarean deliveries (odds ratio 4.96 [95% confidence interval 3.82-6.44], P<.001). This risk increased with assisted delivery and perineal laceration. No statistical difference in the incidence of urinary incontinence was found among women who had elective cesarean deliveries (6.1%), women who had cesarean deliveries after laboring (5.7%), and women who had cesarean deliveries after laboring and pushing (6.4%). CONCLUSION: Urinary incontinence is common in the immediate postpartum period after a woman's first pregnancy. Although vaginal delivery increases the risk of urinary incontinence, labor and pushing alone without vaginal delivery do not appear to increase this risk significantly. LEVEL OF EVIDENCE: II.

Comprehensive definition of human immunodominant CD8 antigens in tuberculosis.

Despite widespread use of the Bacillus Calmette-Guerin vaccine, tuberculosis, caused by infection with Mycobacterium tuberculosis, remains a leading cause of morbidity and mortality worldwide. As CD8+ T cells are critical to tuberculosis host defense and a phase 2b vaccine trial of modified vaccinia Ankara expressing Ag85a that failed to demonstrate efficacy, also failed to induce a CD8+ T cell response, an effective tuberculosis vaccine may need to induce CD8+ T cells. However, little is known about CD8, as compared to CD4, antigens in tuberculosis. Herein, we report the results of the first ever HLA allele independent genome-wide CD8 antigen discovery program. Using CD8+ T cells derived from humans with latent tuberculosis infection or tuberculosis and an interferon-γ ELISPOT assay, we screened a synthetic peptide library representing 10% of the Mycobacterium tuberculosis proteome, selected to be enriched for Mycobacterium tuberculosis antigens. We defined a set of immunodominant CD8 antigens including part or all of 74 Mycobacterium tuberculosis proteins, only 16 of which are previously known CD8 antigens. Immunogenicity was associated with the degree of expression of mRNA and protein. Immunodominant antigens were enriched in cell wall proteins with preferential recognition of Esx protein family members, and within proteins comprising the Mycobacterium tuberculosis secretome. A validation study of immunodominant antigens demonstrated that these antigens were strongly recognized in Mycobacterium tuberculosis-infected individuals from a tuberculosis endemic region in Africa. The tuberculosis vaccine field will likely benefit from this greatly increased known repertoire of CD8 immunodominant antigens and definition of properties of Mycobacterium tuberculosis proteins important for CD8 antigenicity.

Quality of life in women with postpartum anal incontinence.

OBJECTIVE: To estimate the effect of anal incontinence on postpartum quality of life (QOL). METHODS: We conducted a longitudinal follow-up study of women reporting anal incontinence on a statewide postpartum survey to estimate the effect of anal incontinence on QOL up to 2 years postpartum. Quality of life was assessed every 6 months for 2 years based on the Uebersax incontinence impact questionnaire. The effect of severe anal incontinence on QOL was determined by multivariate logistic regression. RESULTS: A total of 1,050 women reported persistent anal incontinence and returned at least two completed QOL surveys during the 2-year study period. Among women with anal incontinence, 51% reported frustration caused by anal incontinence, 26% reported that anal incontinence affected their emotional health, 18.5% reported anal incontinence affected their child-caring abilities, and 16.2% reported a negative effect on social activities. One in three women with anal incontinence reported severe symptoms. Compared with women with mild anal incontinence, women with severe anal incontinence were four to seven times as likely to report that anal incontinence affected emotional health and the ability to perform daily activities. However, only 10% of women sought medical help for anal incontinence at 6 months, 13.5% at 1 year, and 16.7% at 2 years. CONCLUSION: Even 2 years after delivery, more than a quarter of women who experience a new onset of anal incontinence after childbirth report persistently negative QOL. Despite this, few women discuss anal incontinence with their medical providers. LEVEL OF EVIDENCE: III.