The antiviral protein viperin inhibits HCV replication via interaction with NS5A

The interferon-stimulated gene viperin has been shown to have antiviral activity against hepatitis C virus (HCV) in the context of the HCV replicon, although the molecular mechanisms responsible are not well understood. Here we demonstrate that viperin plays an integral part in the ability of interferon to limit replication of cell culture derived HCV (JFH-1) that accurately reflects the complete viral life cycle. Using confocal microscopy and Fluorescence Resonance Energy Transfer (FRET) analysis we demonstrate that viperin localizes and interacts with HCV NS5A at the lipid droplet interface. In addition viperin also associates with NS5A and the pro-viral cellular factor, VAP-A at the HCV replication complex. The ability of viperin to limit HCV replication was dependent on residues within the C-terminus as well as an N-terminal amphipathic helix. Removal of the amphipathic helix redirected viperin from the cytosolic face of the ER and the lipid droplet to a homogenous cytoplasmic distribution, coinciding with a loss of antiviral effect. C-terminal viperin mutants still localized to the lipid droplet interface and replication complexes but did not interact with NS5A proteins as determined by FRET analysis. In conclusion we propose that viperin interacts with NS5A and the host factor VAP-A to limit HCV replication at the replication complex. This highlights the complexity of host control of viral replication by interferon stimulated gene expression

Health and social care of home-dwelling frail older adults in Switzerland : a mixed methods study

Home-dwelling frail older adults are often faced with multimorbidity and complex care needs, requiring health and social care systems that support frail older adults to age in place. The objective of this paper was to investigate the types of formal health and social care as well as informal care and social support used by home-dwelling frail older adults; whether they perceive their support as sufficient; and their experience with and preferences for care and support

Generation and characterisation of transgenic zebrafish expressing human alpha-1-antitrypsin

The Z variant of α1-antitrypsin (Z-AAT) is the most common “severe” deficiency allele of α1-antitrypsin. It affects approximately 2% to 5% of Caucasians of European descent [1-3]. The Z mutation propels the intracellular linkages between Z-AAT monomers forming polymers which results in high levels of retention within the endoplasmic reticulum of hepatocytes [4]. This hepatic accumulation has been associated with the development of Z-AAT-induced liver disease in some PiZZ patients. A collection of different tools is needed to help in the understanding of the exact role of α1-antitrypsin polymers in the development of Z-AAT-induced liver disease. In this thesis, mAbs specific to Z-AAT polymers were generated by using yeast derived Z-AAT as an antigen for injection into rats. The process of mAbs generation and screening led to the identification and characterisation of two unique mAbs 23A5 and 26D5. In particular, mAb 23A5 was found to preferentially bind to α1-antitrypsin polymers. The mAb 23A5 possibly binds a different epitope to mAb 2C1 that has been shown to recognise Z-AAT polymers formed in vivo. mAb 23A5 works in a range of applications such as indirect immunofluorescence, native polyacrylamide gel electrophoresis (PAGE) immunoblotting, denaturing PAGE immunoblotting, immunoprecipitation and immunohistochemistry. A zebrafish liver (ZFL) cell line expressing α1-antitrypsin was also developed in this thesis. Z-AAT in ZFL cells showed delayed secretion compared to wild type α1-antitrypsin (AAT). The expression of Z-AAT was associated with the development of cellular bodies in the cytoplasm of the ZFL cells. There was an increase in the number of ZFL cells displaying the cytoplasmic cellular bodies after heat treatment indicating that the cytomegalovirus promoter used to drive expression of α1-antitrypsin is responsive towards heat. There was evidence of Z-AAT polymers intracellularly and extracellularly in Z-AAT expressing ZFL cells. This indicates the formation of pathological polymers of α1-antitrypsin occurring in the ZFL cell system. Last, a transgenic α1-antitrypsin zebrafish model was developed to study the effects of hepatic accumulation of Z-AAT. The transgenic Z-AAT zebrafish develop “disease” phenotypes related to glycogen storage. The “glycogen” phenotypes were associated with high levels of hepatic glycogen and formation of vacuoles containing glycogen within the cytoplasm of the hepatocytes. Heat treatment of the Z-AAT transgenic zebrafish led to a change in the distribution pattern of hepatic Z-AAT resulting in a more perinuclear and cytoplasmic localisation, suggesting increased Z-AAT retention. There are two key findings from the α1-antitrypsin zebrafish model. First, there is evidence to suggest that transgenic α1-antitrypsin zebrafish maintain hepatic human AAT better than Z-AAT. Therefore, understanding the mechanism might be useful for therapeutic purposes for PiZZ patients. Second, variability was observed in Z-AAT transgenic zebrafish in that not all the fish developed the “disease” phenotypes. This observation is consistent with the PiZZ human condition where not all the patients develop Z-AAT-induced liver disease. Therefore, the transgenic α1-antitrypsin zebrafish model is a suitable model to study the effects of hepatic Z-AAT accumulation

A Contextual Analysis and Logic Model for Integrated Care for Frail Older Adults Living at Home: The INSPIRE Project

Implementation science methods and a theory-driven approach can enhance the understanding of whether, how, and why integrated care for frail older adults is successful in practice. In this study, we aimed to perform a contextual analysis, develop a logic model, and select preliminary implementation strategies for an integrated care model in newly created information and advice centers for older adults in Canton Basel-Landschaft, Switzerland.; We conducted a contextual analysis to determine factors which may influence the integrated care model and implementation strategies needed. A logic model depicting the overall program theory, including inputs, core components, outputs and outcomes, was designed using a deductive approach, and included stakeholders' feedback and preliminary implementation strategies.; Contextual factors were identified (e.g., lack of integrated care regulations, existing community services, and a care pathway needed). Core components of the care model include screening, referral, assessment, care plan creation and coordination, and follow-up. Outcomes included person-centred coordinated care experiences, hospitalization rate and symptom burden, among others. Implementation strategies (e.g., nurse training and co-developing educational materials) were proposed to facilitate care model adoption.; Contextual understanding and a clear logic model should enhance the potential for successful implementation of the integrated care model

A Randomized Controlled Trial of Soy Isoflavone Intake on Mammographic Density among Malaysian Women

Soy intake is associated with lower breast cancer risk in observational studies concerning Asian women, however, no randomized controlled trials (RCT) have been conducted among Asian women living in Asia. This three-armed RCT assessed the effects of one-year soy isoflavone (ISF) intervention on mammographic density (MD) change among healthy peri- and postmenopausal Malaysian women. This study was registered at ClinicalTrials.gov (NCT03686098). Participants were randomized into the 100 mg/day ISF Supplement, 50 mg/day ISF Diet, or control arm, and assessed for change in absolute and relative dense area from digital mammograms conducted at enrolment and after 12 months, compared over time across study arms using Kruskal-Wallis tests. Out of 118 women enrolled, 91 women completed the intervention, while 27 women (23%) were lost in follow up. The ISF supplement arm participants observed a larger decline in dense area (−1.3 cm2), compared to the ISF diet (−0.5 cm2) and control arm (−0.8 cm2), though it was not statistically significant (p = 0.48). Notably, among women enrolled within 5 years of menopause; dense area declined by 6 cm2 in the ISF supplement arm, compared to 2 in the control arm (p = 0.13). This RCT demonstrates a possible causal association between soy ISF intake and MD, a biomarker of breast cancer risk, among Asian women around the time of menopause, but these findings require confirmation in a larger trial

Evaluation of stool short chain fatty acids profiles in the first year of life with childhood atopy-related outcomes

Introduction: Short chain fatty acids (SCFAs) are the main intestinal intermediate and end products of metabolism of dietary fibers/polyphenols by the gut microbiota. The aim of this study was to evaluate the biological implication of stool SCFA profiles determined in the first year of life on the clinical presentation of allergic outcomes in childhood. Methods: From the Growing Up in Singapore Toward healthy Outcomes (GUSTO) cohort, a sub-cohort of 75 participants was recruited. Scheduled questionnaire data was collected for cumulative prevalence of physician-diagnosed eczema, wheezing with the use of nebuliser, and allergen sensitization till the age of 8 years. Stool samples collected at week 3 and months 3, 6 and 12 were quantitated for 9 SCFAs using LC/MS/MS. SCFA data were grouped into lower (below the 25th) and higher (above the 75th percentiles) categories. Generalized Linear Mixed Models was employed to analyse longitudinal association between SCFAs and atopy-related outcomes. Results: Children with lower stool butyric acid levels (≤25th percentile) over the first 3 time points had higher odds ratio (OR) for wheezing (adjOR = 14.6), eczema (adjOR = 13.2), food sensitization (adjOR = 12.3) and combined outcomes of both wheezing and eczema (adjOR = 22.6) till age 8 years, compared to those with higher levels (≥75 percentile). Additionally, lower longitudinal levels of propionic acid (≤25th percentile) over 4 time points in first year of life was associated with recurrent wheezing (≥2 episodes) till 8 years (adjOR = 7.4) (adj p < 0.05). Conclusion: Our results suggest that relatively low levels of gut SCFAs in early life are associated with increased susceptibility to atopic-related outcomes in childhood