Conjugation of hydrophobic moieties enhances potency of antisense oligonucleotides in the muscle of rodents and non-human primates.

We determined the effect of attaching palmitate, tocopherol or cholesterol to PS ASOs and their effects on plasma protein binding and on enhancing ASO potency in the muscle of rodents and monkeys. We found that cholesterol ASO conjugates showed 5-fold potency enhancement in the muscle of rodents relative to unconjugated ASOs. However, they were toxic in mice and as a result were not evaluated in the monkey. In contrast, palmitate and tocopherol-conjugated ASOs showed enhanced potency in the skeletal muscle of rodents and modest enhancements in potency in the monkey. Analysis of the plasma-protein binding profiles of the ASO-conjugates by size-exclusion chromatography revealed distinct and species-specific differences in their association with plasma proteins which likely rationalizes their behavior in animals. Overall, our data suggest that modulating binding to plasma proteins can influence ASO activity and distribution to extra-hepatic tissues in a species-dependent manner and sets the stage to identify other strategies to enhance ASO potency in muscle tissues.

Hepatotoxicity of high affinity gapmer antisense oligonucleotides is mediated by RNase H1 dependent promiscuous reduction of very long pre-mRNA transcripts.

High affinity antisense oligonucleotides (ASOs) containing bicylic modifications (BNA) such as locked nucleic acid (LNA) designed to induce target RNA cleavage have been shown to have enhanced potency along with a higher propensity to cause hepatotoxicity. In order to understand the mechanism of this hepatotoxicity, transcriptional profiles were collected from the livers of mice treated with a panel of highly efficacious hepatotoxic or non-hepatotoxic LNA ASOs. We observed highly selective transcript knockdown in mice treated with non-hepatotoxic LNA ASOs, while the levels of many unintended transcripts were reduced in mice treated with hepatotoxic LNA ASOs. This transcriptional signature was concurrent with on-target RNA reduction and preceded transaminitis. Remarkably, the mRNA transcripts commonly reduced by toxic LNA ASOs were generally not strongly associated with any particular biological process, cellular component or functional group. However, they tended to have much longer pre-mRNA transcripts. We also demonstrate that the off-target RNA knockdown and hepatotoxicity is attenuated by RNase H1 knockdown, and that this effect can be generalized to high affinity modifications beyond LNA. This suggests that for a certain set of ASOs containing high affinity modifications such as LNA, hepatotoxicity can occur as a result of unintended off-target RNase H1 dependent RNA degradation.

The future of nuclear power: value orientations and risk perception.

Since the turn of the 21st century, there has been a revival of interest in nuclear power. Two decades ago, the expansion of nuclear power in the United States was halted by widespread public opposition as well as rising costs and less than projected increases in demand for electricity. Can the renewed enthusiasm for nuclear power overcome its history of public resistance that has persisted for decades? We propose that attitudes toward nuclear power are a function of perceived risk, and that both attitudes and risk perceptions are a function of values, beliefs, and trust in the institutions that influence nuclear policy. Applying structural equation models to data from a U.S. national survey, we find that increased trust in the nuclear governance institutions reduces perceived risk of nuclear power and together higher trust and lower risk perceptions predict positive attitudes toward nuclear power. Trust in environmental institutions and perceived risks from global environmental problems do not predict attitudes toward nuclear power. Values do predict attitudes: individuals with traditional values have greater support for, while those with altruistic values have greater opposition to, nuclear power. Nuclear attitudes do not vary by gender, age, education, income, or political orientation, though nonwhites are more supportive than whites. These findings are consistent with, and provide an explanation for, a long series of public opinion polls showing public ambivalence toward nuclear power that persists even in the face of renewed interest for nuclear power in policy circles.

Impact of chronic liver disease and cirrhosis on health utilities using SF-6D and the health utility index.

Assessment of health-related quality of life (HRQL) and health utilities have become important aspects of clinical research. Patient-derived utility adjustments are frequently used in economic analysis. Although HRQL has been frequently studied among patients with liver disease, extensive data on the health utilities of patients with liver disease are not available. Recently, SF-6D has been developed to obtain utility scores from the widely used Short Form 36 questionnaire. To assess health utilities of patients with chronic liver disease using 2 utility assessments [SF-6D and Health Utility Index 2 (HUI-2)], a total of 140 patients were identified from our Liver Disease Quality of Life Database with HRQL data available, as well as clinical and demographic data. Of the 140 patients, 42% were female, had a mean age of 49.4 years (standard deviation = +/-11.2) 36% had hepatitis B virus (HBV), 29% had hepatitis C (HCV), 24% had cholestatic liver disease, and 11% had another liver disease (for example, nonalcoholic steatohepatitis). Bivariate analyses indicated that HBV patients had the highest health status as measured by all of SF-6D and HUI-2 subscales and the overall SF-6D and HUI-2 utility measures, whereas patients with HCV and cholestatic liver disease had similar scores, and those with other liver diseases had the poorest quality of life. When controlling for the effects of gender, age, and cirrhosis, impact of chronic liver disease diagnosis on utility scores persisted only for the SF-6D, with HCV patients having significantly poorer health than HBV patients. In conclusion, SF-6D provides not only a generic assessment of HRQL but also a utility score that can be used for economic analysis of patients with chronic liver disease.

Health-related quality of life in patients with chronic hepatitis B.

UNLABELLED: Although chronic hepatitis C (CH-C) has consistently been shown to impair patients' health-related quality of life (HRQL), the impact of chronic hepatitis B (CH-B) on HRQL has not been fully explored. AIM: Compare HRQL between patients with CH-B, CH-C, primary biliary cirrhosis (PBC) and healthy controls. DESIGN: Three HRQL questionnaires [Chronic Liver Disease Questionnaire (CLDQ), Short Form 36 (SF-36) and the Health Utility Index (HUI Mark-2 and Mark-3)] were administered prospectively. Additional clinical and laboratory data and normative data for healthy individuals, were available. ANALYSIS: Scores were compared using analysis of variance and multiple regression. RESULTS: One hundred and forty-six patients with CH-B, CH-C and PBC were included [mean age 47.1 years (+/-11.6), 41% female, 33% cirrhosis]. CH-C and PBC patients scored the lowest on all CLDQ, SF-36 and HUI domains compared with CH-B patients and healthy controls. CH-B patients had scores similar to the healthy population, measured by most CLDQ and SF-36 scales. However, the HUI scores for CH-B patients showed more impairment than population norms. Having CH-B and not having cirrhosis were predictive of utility and HRQL scores in multivariate models. CONCLUSIONS: CH-B patients have better HRQL than CH-C, PBC and population norms. CH-B patients' overall utility scores are lower than population norms.

Anger experiences among hepatitis C patients: relationship to depressive symptoms and health-related quality of life.

The authors examined anger among hepatitis C (HCV) patients and its relationship to health-related quality of life (HRQL) and depression. Eighty-seven HCV patients who received pegylated interferon-alpha(2b) and ribavirin were included. Patients' mean age was 48 years; 42% were women, and 60% were white. Patients experienced moderate anger while undergoing HCV treatment. Angry feelings increased during treatment in some domains, specifically, Control Over Anger and Angry Reaction. Greater anger was associated with more depression and poorer HRQL. Findings point to the importance for physicians to screen for a wide range of neuropsychiatric side effects of interferon, including anger.

Development of siRNA for therapeutics: efficient synthesis of phosphorothioate RNA utilizing phenylacetyl disulfide (PADS).

Efficient synthesis of phosphorothioate RNA (PS-RNA) is demonstrated by using phenylacetyl disulfide (PADS) in a mixture of pyridine and acetonitrile (1:1, v/v) for 3 min. Sulfurization is achieved with >99.8% stepwise efficiency. This reagent also performs efficiently during synthesis of RNA containing PS:PO mixed backbone.

Depression, anemia and health-related quality of life in chronic hepatitis C.

BACKGROUND/AIMS: Hepatitis C (HCV) infected patients have significant health-related quality of life (HRQL) impairment which worsens during anti-viral therapy. Our aim was to examine the association of HRQL with treatment-induced depression and anemia. METHODS: Two hundred and seventy-one HCV patients who received pegylated interferon alfa 2b and ribavirin were included. Data on HRQL, depressive symptoms, laboratory values and socio-demographic characteristics were collected. RESULTS: Mean age was 47.1+/-6.5, 69% were male, and 73% were White. HCV patients' HRQL declined during anti-viral therapy but returned to or exceeded baseline levels within 24 weeks of completion. Anemia and depression were both associated with HRQL impairment. The effects of depression on HRQL were strong; once depression scores were included other factors were no longer significant. Patients' depressive symptoms tended to increase during the initial half of treatment regimen. Those with higher body mass index (BMI), cirrhosis, and women reported more HRQL impairments. HRQL scales were generally not associated with alcohol abuse, age, race, ALT and HCV RNA levels. CONCLUSIONS: Anti-viral therapy for HCV is associated with diminished HRQL. Although anemia and depression were associated with this impairment, depression was the most consistent predictor. Future studies are needed to see whether proactive management of these side effects can improve patients' HRQL and the efficacy of antiviral therapy for hepatitis C.

The personal and social context of planning for end-of-life care.

OBJECTIVES: To examine the potential facilitators of or deterrents to end-of-life planning for community-dwelling older adults, including personal (health-related and sociodemographic) and social (physician and family) influences. DESIGN: In-person interviews with older adults, telephone interviews with physicians and family members. SETTING: Cleveland, Ohio. PARTICIPANTS: Two hundred thirty-one adults aged 65 to 99 who were aging in place, 99 of their primary care physicians, and 127 of their family members. MEASUREMENTS: Questions assessing older adults' discussions with others about end-of-life plans, implementation of advance directives, and physical (Older American Resources and Services) illness index and mental (Short Portable Mental Status Questionnaire) health status. RESULTS: Just fewer than half of older adults had executed an advance directive and discussed their wishes with others. Only personal characteristics of elderly individuals were related to end-of-life plans, with whites, unmarried individuals, and younger adults more likely to have made preparations. Older adults' health status, as evaluated by the patient, physician, and caregiver, did not relate to the tendency to have made advance care plans. Older adults' family members were much more likely to report knowledge of advance care plans than were physicians. CONCLUSION: These findings suggest that many physicians are not talking with their patients about their end-of-life wishes. Furthermore, the propensity to have such discussions may relate more to the personal preferences and level of comfort of patients, physicians, and family members than on the health status of the older adult.