Predictors of health-related quality of life among military HIV-infected individuals.

OBJECTIVES: To determine long-term predictors of health-related quality of life (HRQOL) and evaluate the treatment effect of highly active antiretroviral therapy (HAART) on HRQOL in the US Military HIV Natural History Study (NHS) cohort. METHODS: Participants were a nested cohort of the NHS who responded to the Rand Short Form 36 questionnaire administered from 2006 to 2010. Physical component summary scores (PCS) and mental component summary scores (MCS) were computed using standard algorithms. HAART-status was categorized as non-protease inhibitor-based (NPI-HAART), protease inhibitor-based (PI-HAART), HAART-naïve, or off-HAART. Mixed linear random effects models were used to estimate changes in PCS and MCS over time for treatment and covariates (including CD4 count, HIV viral load, medical and mental comorbidities). RESULTS: Eight hundred and twelve participants met the inclusion criteria. There was no difference in PCS or MCS between those on PI-HAART compared to NPI-HAART. Significant predictors of PCS were CD4 count < 200 cells/mm3 (ß = - 2.90), CD4 count 200-499 cells/mm3 (ß = - 0.80), and mental comorbidity (ß = - 3.23). Others were medical comorbidity, AIDS-defining illness, being on NPI-HAART, HAART-naïve, age, and rank. Those with medical comorbidities experienced yearly improvement in PCS. Predictors of MCS were CD4 count < 200 cells/mm3 (ß = - 2.53), mental comorbidity (ß = - 4.58), and being African American (ß = 2.59). CONCLUSION: HRQOL was significantly affected by low CD4 count, medical and mental comorbidities. Addressing these modifiable factors would be expected to improve the physical and mental HRQOL of the cohort. Our study did not find any treatment benefit of NPI-HAART over PI-HAART on HRQOL in the long term.

Hepatitis B Birth Dose Effects on Childhood Immunization in the U.S.

INTRODUCTION: The Advisory Committee on Immunization Practices recommends administering the first dose of hepatitis B vaccine at birth, making it the first vaccine that many children receive. However, few studies examine whether children who miss the birth dose are at increased risk of vaccination delay. This study investigates birth dose as a determinant of up-to-date immunization status at age 18 months, considering 7 core childhood vaccine series: diphtheria, tetanus, and acellular pertussis; polio; measles, mumps, and rubella; Haemophilus influenzae type B; varicella; hepatitis B; and pneumococcal conjugate vaccine. METHODS: Cross-sectional data were collected in 2017 by National Immunization Survey-Child, a nationally representative survey of children aged 19-35 months living in the U.S., and were analyzed in 2019. The primary outcome was combined 7-vaccine series (4:3:1:3:3:1:4) up-to-date status at 18 months. Doubly robust estimates of association were calculated using survey logistic regression and propensity scores estimated with boosted classification and regression trees. RESULTS: Children who received the birth dose had 2.01 (95% CI=1.74, 2.33) times the odds of being up-to-date on the combined 7-vaccine series as children who did not. ORs for all the 7 individual vaccine series were positive, ranging from 1.59 (95% CI=1.28, 1.97) for measles, mumps, and rubella to 4.97 (95% CI=3.97, 6.24) for hepatitis B. CONCLUSIONS: Receiving the birth dose is positively associated with up-to-date status later in childhood, highlighting the importance of starting vaccination early. The association is insensitive to confounding by factors observed in National Immunization Survey-Child, but investigation of unobserved factors such as vaccine hesitancy could provide critical information to guide intervention strategy.

Lower health-related quality of life predicts all-cause hospitalization among HIV-infected individuals.

BACKGROUND: Health-related quality of life (HRQOL) is a patient-centered outcome measure used in assessing the individual's overall functional health status but studies looking at HRQOL as a predictive tool are few. This work examines whether summary scores of HRQOL are predictive of all-cause hospitalization in the US Military HIV Natural History Study (NHS) cohort. METHODS: The Short Form 36 (SF-36) was administered between 2006 and 2010 to 1711 NHS cohort members whose hospitalization records we had also obtained. Physical component summary scores (PCSS) and mental component summary scores (MCSS) were computed based on standard algorithms. Terciles of PCSS and MCSS were generated with the upper terciles (higher HRQOL) as referent groups. Proportional hazards multivariate regression models were used to estimate the hazard of hospitalization for PCSS and MCSS separately (models 1 and 2, respectively) and combined (model 3). RESULTS: The hazard ratios (HR) of hospitalization were respectively 2.12 times (95% CI: 1.59-2.84) and 1.59 times (95% CI: 1.19-2.14) higher for the lower and middle terciles compared to the upper PCSS tercile. The HR of hospitalization was 1.33 times (95% CI: 1.02-1.73) higher for the lower compared to the upper MCSS tercile. Other predictors of hospitalization were CD4 count < 200 cells/mm3 (HR = 2.84, 95% CI: 1.96, 4.12), CD4 count 200-349 cells/mm3 (HR = 1.67, 95% CI: 1.24, 2.26), CD4 count 350-499 cells/mm3 (HR = 1.41, 95% CI: 1.09, 1.83), plasma viral load > 50 copies/mL (HR = 1.82, 95% CI: 1.46, 2.26), and yearly increment in duration of HIV infection (HR = 0.94, 95% CI: 0.93, 0.96) (model 3). CONCLUSION: After controlling for factors associated with hospitalization among those with HIV, both PCSS and MCSS were predictive of all-cause hospitalization in the NHS cohort. HRQOL assessment using the SF-36 may be useful in stratifying hospitalization risk among HIV-infected populations.

Health-related quality of life among military HIV patients on antiretroviral therapy.

OBJECTIVE: The aims of this study were: (i) to determine the factors associated with HRQOL at baseline in our cohort, and (ii) to evaluate if there are differences in baseline HRQOL measures by antiretroviral treatment. METHODS: The Short Form 36 (SF-36) was administered between 2006 and 2010 among members of the United States HIV Natural History Study cohort (NHS), and participants who completed the SF-36 were included in the study. Physical component summary (PCS) and mental component summary (MCS) scores were computed based on standard algorithms. Multivariate linear regression models were constructed for PCS and MCS to estimate the association between selected variables and HRQOL scores. RESULTS: Antiretroviral therapy (ART) was not independently associated with HRQOL scores. Factors associated with PCS were CD4+ count < 200 cells/mm3 (ß = -5.84, 95% CI: -7.63, -4.06), mental comorbidity (ß = -2.82, 95% CI: -3.79, -1.85), medical comorbidity (ß = -2.51, 95% CI: -3.75, -1.27), AIDS diagnosis (ß = -2.38, 95% CI: -3.79, -0.98). Others were gender, military rank, marital status, and age. Factors independently associated with MCS were CD4+ count < 200 cells/mm3 (ß = -1.93, 95% CI: -3.85, -0.02), mental comorbidity (ß = -6.25, 95% CI: -7.25, -5.25), age (ß = 0.37, 95% CI: 0.14, 0.60), and being African American (ß = 1.55, 95% CI: 0.63, 2.47). CONCLUSION: Among military active duty and beneficiaries with HIV, modifiable factors associated with HRQOL measures included advanced HIV disease, and mental or medical comorbidity. Addressing these factors may improve quality of life of HIV-infected individuals in the NHS cohort.

No contribution of lifestyle and environmental exposures to gender discrepancy of liver disease severity in chronic hepatitis b infection: Observations from the Haimen City cohort.

BACKGROUND: Previous studies have noted significant gender difference in the risk of liver cancer among hepatitis B chronic infection patients. Some indicated that it might be due to lifestyle-related differences. This paper tests whether or not such a gender discrepancy among the chronic hepatitis B population is confounded by lifestyle and environment related exposures. METHODS: We retrieved a sample of 1863 participants from a prospective cohort in Haimen City, China in 2003. Liver disease severity was categorized as "normal", "mild", "moderate", and "severe" based on a clinical diagnosis. Lifestyle and environmental exposures were measured by questionnaires. We used factor analysis and individual variables to represent lifestyle and environmental exposures. We applied the cumulative logit models to estimate the effect of gender on liver disease severity and how it was impacted by lifestyle and environmental exposures. RESULTS: Gender and HBeAg positivity were independent risk factors for more severe liver disease. Compared to females, males were 2.08 times as likely to develop more severe liver disease (95% CI: 1.66-2.61). Participants who were HBeAg positivite were 2.19 times (95% CI: 1.61-2.96) as likely to develop more severe liver disease compared to those who were negative. Controlling for lifestyle and environmental exposures did not change these estimations. CONCLUSIONS: Males in the HBV infected population have an increased risk of severe liver disease. This gender effect is independent of the lifestyle and environmental exposures addressed in this study. Our findings support the hypothesis that gender discrepancies in HCC risk are attributable to intrinsic differences between males and females.

Hepatitis B knowledge among key stakeholders in Haimen City, China: Implications for addressing chronic HBV infection.

BACKGROUND: This article describes hepatitis B-related knowledge, attitudes and practices after completion of the Gateway to Care campaign, a citywide public health education program that targeted city residents, health care providers and individuals chronically infected with hepatitis B virus in Haimen City, China. METHODS: Pre/post questionnaires assessed hepatitis B knowledge change among health care providers and post-campaign surveys evaluated hepatitis B knowledge, attitudes and behaviors (including stigma-related beliefs and practices) among health care providers, city residents and chronically infected individuals. Focus groups were conducted to gain a more in-depth understanding of the needs of the target communities, and to identify future intervention strategies to improve hepatitis B testing and linkage to care and treatment. RESULTS: Results indicate high levels of hepatitis B knowledge among multiple stakeholders in Haimen City, with significant knowledge improvement among health care providers. Stigma-related beliefs and myths regarding separation of infected individuals from certain aspects of family life were common among all stakeholder groups, despite high levels of accurate knowledge about hepatitis B transmission and prevention. Self-report of hepatitis B screening was low among city residents, as was awareness of hepatitis B treatment. CONCLUSIONS: More efforts are needed to improve awareness of HBV treatment, decrease HBV-related stigma, improve screening rates, and reduce cost of antiviral treatment. Future interventions in Haimen City should be driven by behavioral change theory, to not only improve knowledge, but to improve screening behaviors and address hepatitis B-related stigma and discrimination.

Hepatitis-Associated Liver Cancer: Gaps and Opportunities to Improve Care.

The global burden of hepatocellular carcinoma (HCC; primary liver cancer) is increasing. HCC is often unaccompanied by clear symptomatology, causing patients to be unaware of their disease. Moreover, effective treatment for those with advanced disease is lacking. As such, effective surveillance and early detection of HCC are essential. However, current screening and surveillance guidelines are not being fully implemented. Some at-risk populations fall outside of the guidelines, and patients who are screened are often not diagnosed at an early enough stage for treatment to be effective. From March 17 to 19, 2015, the Hepatitis B Foundation sponsored a workshop to identify gaps and limitations in current approaches to the detection and treatment of HCC and to define research priorities and opportunities for advocacy. In this Commentary, we summarize areas for further research and action that were discussed throughout the workshop to improve the recognition of liver disease generally, improve the recognition of liver cancer risk, and improve the recognition that screening for HCC makes a life-saving difference. Participants agreed that primary prevention of HCC relies on prevention and treatment of viral hepatitis and other underlying etiologies. Earlier diagnosis (secondary prevention) needs to be substantially improved. Areas for attention include increasing practitioner awareness, better definition of at-risk populations, and improved performance of screening approaches (ultrasound, biomarkers for detection, risk stratification, targeted therapies). The heterogeneous nature of HCC makes it unlikely that a single therapeutic agent will be universally effective. Medical management will benefit from the development of new, targeted treatment approaches.

Choice of measures of vaccination and estimates of risk of pediatric pertussis.

BACKGROUND: Vaccination uptake at the individual level can be assessed in a variety of ways, including traditional measures of being up-to-date (UTD), measures of UTD that consider dose timing, like age-appropriate vaccination, and risk reduction from individual doses. This analysis compared methods of operationalizing vaccination uptake and corresponding risk of pertussis infection. METHODS: City-wide case-control study of children in Philadelphia aged 3 months through 6 years, between 2001 and 2013. Multiple logistic regression was used to isolate the independent effects of each measure of vaccination uptake and the corresponding relative odds of pertussis. RESULTS: Being UTD on vaccinations was associated with a 52% reduction in risk of pertussis (OR 0.48, 95% CI: 0.34, 0.69). Evaluation of delayed receipt of vaccine versus on-time UTD yielded similar results. There was a decrease in risk of pertussis for each additional dose received with the greatest reduction in pertussis infection observed from the first (OR 0.48, 95% CI: 0.28, 0.83) and second dose (OR 0.17, 95% CI: 0.08, 0.34). Additional doses conferred minimal additional protection in this age group. CONCLUSION: Examining vaccination status by individual doses may offer improved predictive capacity for identifying children at risk for pertussis infection compared to the traditional UTD measure.

Prevention of perinatal hepatitis B transmission in Haimen City, China: Results of a community public health initiative.

In regions where hepatitis B virus (HBV) is endemic, perinatal transmission is common. Infected newborns have a 90% chance of developing chronic HBV infection, and 1 in 4 will die prematurely from HBV-related liver disease. In 2010, the Hepatitis B Foundation and the Haimen City CDC launched the Gateway to Care campaign in Haimen City, China to improve awareness, prevention, and control of HBV infection citywide. The campaign included efforts to prevent perinatal HBV transmission by screening all pregnant women for hepatitis B surface antigen (HBsAg), following those who tested positive, and administering immunoprophylaxis to their newborns at birth. Of 5407 pregnant women screened, 185 were confirmed HBsAg-positive and followed until delivery. At age one, 175 babies were available for follow up testing. Of those, 137 tested negative for HBsAg and positive for antibodies to HBsAg, indicating protection. An additional 34 HBsAg-negative babies also tested negative for antibodies to HBsAg or had indeterminate test results, were considered to have had inadequate immune responses to the vaccine, and were given a booster dose. A higher prevalence of nonresponse to HBV vaccine was observed among babies born to hepatitis B e antigen (HBeAg)-positive mothers and mothers with high HBV DNA titers. The remaining 4 babies tested positive for HBsAg and negative for antibodies, indicative of active HBV infection. The mothers of all 4 had viral loads ≥8×10(6) copies/ml in the third trimester. Although inadequate response or nonresponse to HBV vaccine was more common among babies born to HBeAg-positive and/or high viral load mothers, these risk factors did not completely predict nonresponsiveness. All babies born to HBV-infected mothers should be tested upon completion of the vaccine series to ascertain adequate protection. Some babies of HBeAg-positive mothers with high viral load may still become HBV infected despite timely immunoprophylaxis with HBV vaccine and HBIG.

Gateway to Care campaign: a public health initiative to reduce the burden of hepatitis B in Haimen City, China.

BACKGROUND: An estimated one million people worldwide die each year from complications of chronic hepatitis B infection (CHB), including liver cancer. A disproportionate number of infections and deaths occur in China. The incidence and mortality of liver cancer in Haimen City is among the highest in China, and in the world. A multi-year citywide campaign was aimed at eliminating hepatitis B virus (HBV) infection and significantly reducing the number of liver cancer deaths due to CHB in Haimen City, China. METHODS: Strategies included a public health information campaign targeting the 1.03 million city residents; specialized health education for leaders and providers to increase adoption of evidence-based HBV management protocols; establishment of health care infrastructure and management systems; and increased prevention and care delivery to key subpopulations (especially pregnant women). RESULTS: The project developed and deployed broad-reaching public awareness and health education tools and modules to 280,000 households and at community-based events. More than 90% of targeted healthcare providers and 80% of the community leaders/government officials attended educational seminars during the project period (1,441 health care providers; 1,883 local government officials). A centralized registration and management system for pregnant women was developed and instituted, 100% of pregnant women were enrolled (5,407 women over one year), and all infants born to HBV-infected mothers received one dose of HBIG and the first dose of HBV vaccine by 24 hours of birth. CONCLUSIONS: Lessons from the implementation phase of the project include the importance of: gaining early and ongoing support from the local government and health bureau for success in reaching the targeted populations; and having project management by a local, experienced, and trusted health expert to navigate implementation and relationships, and help develop culturally and linguistically appropriate materials.

Chronic HBV infection outside treatment guidelines: is treatment needed?

BACKGROUND: There are now seven antivirals approved for use in the management of chronic HBV infection in the US. Current professional guidelines recommend the use of antiviral treatment in only a distinct subset of the total HBV chronically infected population, estimated to be more than 350 million worldwide. The subset of chronically HBV-infected individuals for whom the antivirals have been demonstrated to produce desirable outcomes are those with abnormal liver enzymes and a viral load above a defined threshold, presumably identifying those at highest risk for development of cirrhosis and hepatocellular carcinoma. However, some individuals whose clinical features place them outside these guidelines, for whom treatment is not recommended, are also at significant risk for liver disease complications and liver-related death. METHODS: In this report, we produce new estimates of the age-specific risks of liver-related death in people outside the current treatment guidelines using published data from multiple populations. RESULTS: Our results indicate that the age-specific 10-year risks of liver-related mortality in these individuals range from 0.3-4% in the West to 0.3-20% in Asia. CONCLUSIONS: The magnitude of these risks and the estimated size of the global population that falls outside of current treatment guidelines have led us to consider whether medical interventions are also needed for these individuals, either with currently approved therapeutics or yet-to-be-discovered medications targeting new mechanisms of antiviral effect. Potential targets for development of new medications are discussed.

Comparative effectiveness of empiric ß-lactam monotherapy and ß-lactam-macrolide combination therapy in children hospitalized with community-acquired pneumonia.

OBJECTIVE: To determine the comparative effectiveness of ß-lactam monotherapy and ß-lactam and macrolide combination therapy on clinical outcomes in the treatment of children hospitalized with community-acquired pneumonia (CAP). STUDY DESIGN: This multicenter retrospective cohort study included children aged 1-18 years who were hospitalized with CAP and received ß-lactam antibiotic therapy either alone or in combination with a macrolide. Data were obtained from the Pediatric Health Information System. Associations between empiric antibiotic therapy and hospital readmission for the same episode of pneumonia were estimated using exact logistic regression. Associations between empiric antibiotic therapy and length of hospital stay were estimated using a generalized estimating equation with negative binomial distribution. RESULTS: There were 20 743 patients hospitalized with CAP. Of these, 24% received ß-lactam and macrolide combination therapy on admission. Compared with children who received ß-lactam monotherapy, children who received ß-lactam plus macrolide combination therapy were 20% less likely to stay in the hospital an additional day (adjusted relative risk 0.80; 95% CI, 0.75-0.86) but did not have a different readmission rate (relative risk 0.69; 95% CI, 0.41-1.12). An effect of combination treatment on reduced length of stay was not evident in children <6 years of age but increased with increasing age groups thereafter. CONCLUSION: School-aged patients hospitalized with CAP who received ß-lactam plus macrolide combination therapy have a shorter length of stay and similar rates of readmission compared with school-aged patients who receive ß-lactam monotherapy.

Dichlorodiphenyltrichloroethane and risk of hepatocellular carcinoma.

Dichlorodiphenyltrichloroethane (p,p'-DDT), an organochlorine pesticide known to have deleterious health effects in humans, has been linked to hepatocellular carcinoma (HCC) in rodents. A recent study has reported that p,p'-DDT and its most persistent metabolite, dichlorodiphenyldichloroethylene (p,p'-DDE), may also be associated with HCC in humans. To examine whether there is an association between p,p'-DDT and/or p,p'-DDE in a population at high-risk of developing HCC, a nested case-control study was conducted within the 83,794 person Haimen City Cohort in China. Sera and questionnaire data were collected from all participants between 1992 and 1993. This study included 473 persons who developed HCC and 492 who did not, frequency matched on sex, age and area of residence. p,p'-DDT and p,p'-DDE levels were determined by mass spectrometry. Hepatitis B viral infection status (based on hepatitis B virus surface antigen; HBsAg) was also determined. p,p'-DDT and/or p,p'-DDE serum levels were significantly associated with sex, area of residence, occupation, alcohol consumption and cigarette smoking. Adjusting for age, sex, area of residence, HBsAg, family history of HCC, history of acute hepatitis, smoking, alcohol, occupation (farmer vs. other) and levels of p,p'-DDT or p,p'-DDE, odds ratios (OR) and 95% confidence intervals (CI) were calculated via unconditional logistic regression. Overall, the highest quintile of p,p'-DDT was associated with an increased risk of HCC, OR = 2.96 95% CI; 1.19-7.40. There were no statistically significant associations with p,p'-DDE. Overall, these results suggest that recent exposure to p,p'-DDT may increase risk of HCC.

Quantitative dynamics of hepatitis B basal core promoter and precore mutants before and after HBeAg seroconversion.

BACKGROUND & AIMS: Hepatitis B e antigen (HBeAg) seroconversion is an important clinical and virological "landmark" during chronic hepatitis B virus (HBV) infection. Mutant viruses carrying the precore G1896A and/or the basal core promoter (BCP) A1762T/G1764A mutations are associated with HBeAg seroconversion. However, the exact role of these mutants in HBeAg seroconversion remains unclear, partly because the evolution of these mutant viruses before and after seroconversion has not been well studied. METHODS: Using our novel mutant quantification methods, the percentage of the mutant viruses was analyzed both cross-sectionally and longitudinally, before and after seroconversion. RESULTS: Cross-sectional analysis showed that the percentage of both precore and BCP mutants gradually increased with age in the HBeAg-positive population. Follow-up of 18 HBeAg-positive patients revealed that the mutant percentage may stay low and stable for many years, followed by a steady increase in the percentage of G1896A and/or A1762T/G1764A mutants, from <10% to 50-100%, within about 3 years prior to seroconversion. In all cases, increase of mutant percentage was preceded or accompanied by elevated serum alanine aminotransferase. After the seroconversion, the mutant percentage could remain high or decrease significantly, sometimes to below 20%. CONCLUSIONS: Levels of G1896A and A1762T/G1764A mutants (of genotypes B and C) in the HBeAg-positive patients may predict the time of HBeAg seroconversion. The dominance of these mutants in the HBeAg-positive phase is more likely the result of immune selection rather than the enhanced replication capability of the mutants. However, anti-HBe antibody may not be a major selection force for these mutants.

Fumonisin B1 and risk of hepatocellular carcinoma in two Chinese cohorts.

Fumonisin B1 (FB1), a mycotoxin that contaminates corn in certain climates, has been demonstrated to cause hepatocellular cancer (HCC) in animal models. Whether a relationship between FB1 and HCC exists in humans is not known. To examine the hypothesis, we conducted case-control studies nested within two large cohorts in China; the Haimen City Cohort and the General Population Study of the Nutritional Intervention Trials cohort in Linxian. In the Haimen City Cohort, nail FB1 levels were determined in 271 HCC cases and 280 controls. In the General Population Nutritional Intervention Trial, nail FB1 levels were determined in 72 HCC cases and 147 controls. In each population, odds ratios and 95% confidence intervals (95%CI) from logistic regression models estimated the association between measurable FB1 and HCC, adjusting for hepatitis B virus infection and other factors. A meta-analysis that included both populations was also conducted. The analysis revealed no statistically significant association between FB1 and HCC in either Haimen City (OR=1.10, 95%CI=0.64-1.89) or in Linxian (OR=1.47, 95%CI=0.70-3.07). Similarly, the pooled meta-analysis showed no statistically significant association between FB1 exposure and HCC (OR=1.22, 95%CI=0.79-1.89). These findings, although somewhat preliminary, do not support an associated between FB1 and HCC.

Methylation of the CpG sites only on the sense strand of the APC gene is specific for hepatocellular carcinoma.

Hypermethylation of the promoter of the tumor suppressor gene, adenomatous polyposis coli (APC), occurs in various malignancies, including hepatocellular carcinoma (HCC). However, reports on the specificity of the methylation of the APC gene for HCC have varied. To gain insight into how these variations occur, bisulfite PCR sequencing was performed to analyze the methylation status of both sense and antisense strands of the APC gene in samples of HCC tissue, matched adjacent non-HCC liver tissue, hepatitis, cirrhosis, and normal liver tissues. DNA derived from fetal liver and 12 nonhepatic normal tissue was also examined. These experiments revealed liver-specific, antisense strand-biased CpG methylation of the APC gene and suggested that, although methylation of the antisense strand of the APC gene exists in normal liver and other non-HCC disease liver tissue, methylation of the sense strand of the APC gene occurs predominantly in HCC. To determine the effect of the DNA strand on the specificity of the methylated APC gene as a biomarker for HCC detection, quantitative methylation-specific PCR assays for sense and antisense strand DNA were developed and performed on DNA isolated from HCC (n = 58), matched adjacent non-HCC (n = 58), cirrhosis (n = 41), and hepatitis (n = 39). Receiver operating characteristic curves were constructed. With the cutoff value set at the limit of detection, the specificity of sense and antisense strand methylation was 84% and 43%, respectively, and sensitivity was 67.2% and 72.4%, respectively. This result demonstrated that the identity of the methylated DNA strand impacted the specificity of APC for HCC detection. Interestingly, methylation of the sense strand of APC occurred in 40% of HCCs from patients with serum AFP levels less than 20 ng/mL, suggesting a potential role for APC as a biomarker to complement AFP in HCC screening.

Retail food safety risks for populations of different races, ethnicities, and income levels.

Research has found that populations with low socioeconomic status (SES) and minority populations have greater access to small corner markets and less access to supermarkets than high-SES and Caucasian populations. This represents a significant difference in the farm-to-fork continuum that these populations experience. This research examined whether differential retail access to foods results in different food safety risks at the retail level for consumers with different demographics. U.S. Census Bureau census tracts with high African American, Asian, Hispanic, Caucasian, low-SES, and high-SES populations were identified in Philadelphia, PA. Approximately 60 retail food establishments were sampled in each census tract category from June 2008 to June 2010. Food samples collected at stores included milk, eggs, lunchmeat, sandwiches, and ready-to-eat (RTE) fresh fruit, greens, and herbs, when available. With the exception of milk and eggs, only food that had been handled and/or prepared at the retail level was sampled. Food samples were tested for temperature, aerobic plate count, coliforms, fecal coliforms, Escherichia coli, Staphylococcus aureus, and Listeria monocytogenes. The results indicated that internal egg temperatures were higher in samples from low-SES census tracts than in eggs from Caucasian census tracts, and eggs were more often found unrefrigerated in markets in low-SES and Asian census tracts. Milk samples from markets in Hispanic and low-SES census tracts had higher aerobic plate counts than high-SES census tract samples. Sandwiches from markets in high-SES census tracts had higher coliform counts than sandwiches from markets in all other census tract categories. Markets in Asian census tracts had a higher incidence of fecal coliform contamination on sandwiches than markets in Caucasian census tracts. Fecal coliforms were present in a percentage of RTE greens from markets in all census tracts except African American, with the highest percentages of RTE greens positive for fecal coliforms in low-SES (100%), Asian (71.4%), and Caucasian (45.5%) markets.

Quantification of complex precore mutations of hepatitis B virus by SimpleProbe real time PCR and dual melting analysis.

BACKGROUND: Hepatitis B virus (HBV) precore G1896A mutation is associated with Hepatitis B e antigen (HBeAg) seroconversion. This mutation and the adjacent G1899A mutation also appear to associate with increased risk of hepatocellular carcinoma. Quantitative mutant dynamics may help determine the potential of these mutants as clinical biomarkers. However, a reliable method to quantify either mutant is not available, partly because the viral genome has polymorphisms in general and the precore mutations are complex. OBJECTIVES: (1) To develop a reliable and ultrasensitive assay for the quantification of HBV G1896A and/or G1899A mutants. (2) To obtain preliminary data on the quantities of the precore mutants in patients. STUDY DESIGN: A SimpleProbe real time PCR assay was developed to quantify the HBV precore mutants. Dual melting analysis and a primer-probe partial overlap approach were used to increase detection accuracy. A wild-type selective PCR blocker was also developed to increase mutant detection sensitivity. RESULTS: The assay correctly identified the precore sequence from all 62 patient samples analyzed. More than 97% of precore sequences in the GenBank can be recognized. Mutant detection sensitivity reached 0.001% using a wild type-selective PCR blocker. At least one precore mutant can be detected from all 20 HBeAg-positive individuals who were negative for precore mutations by DNA sequencing. CONCLUSIONS: The reliability of this ultrasensitive mutation quantification assay was demonstrated. The same approaches may be useful for the detection of other clinically significant mutations. Evolution of the precore mutants warrants further studies.