Methods: The NanoLuc™ Luciferase reporter protein was engineered to be expressed as a fusion protein for MNV-1 minor capsid protein, VP2. The foot-and-mouth disease virus 2A (FMDV2A) sequence was inserted between the 3'end of the reporter gene and the VP2 start sequence to allow co-translational 'cleavage' of fusion proteins during intracellular transcript expression. Amplification of the fusion gene was performed using a series of standard and overlapping polymerase chain reactions. The resulting amplicon was then cloned into three readily available backbones of MNV-1 cDNA clones.
Results: Restriction enzyme analysis indicated that the NanoLucTM Luciferase gene was successfully inserted into the parental MNV-1 cDNA clone. The insertion was further confirmed by using DNA sequencing.
Conclusion: NanoLuc™ Luciferase-tagged MNV-1 cDNA clones were successfully engineered. Such clones can be exploited to develop robust experimental assays for in vitro assessments of viral RNA replication.
METHOD: A multicentre, parallel, randomised, controlled, open-label, non-inferiority trial was conducted. Adult patients receiving CAPD were randomised in a 1:1 ratio to SSL or STS. The primary outcome was the rate of peritonitis after 1 year of follow-up.
RESULTS: A total of 472 subjects were randomised (SSL, n = 233; STS, n = 239). One subject in each group was excluded from the analysis as they withdrew consent before the first dialysis dose. Four hundred and seventy subjects (SSL, n = 232; STS, n = 238) were included in the modified intention-to-treat analysis. Non-inferiority between two groups was established as no significant difference was found in peritonitis rate (incident rate ratio: 0.91, 95% CI: 0.65-1.28). No significant difference was detected in weekly Kt/V (p = 0.58) and creatinine clearance (p = 0.55). However, the average ultrafiltration volume was significantly lower in SSL, with a mean difference of 93 ml (p < 0.01). SSL also demonstrated a 2.57-times higher risk of device defect than STS (95% CI: 1.77-3.75).
CONCLUSION: SSL was non-inferior in peritonitis rate compared to plastic-free STS over 1 year in patients requiring CAPD. There was no difference in the delivered dialysis dose, but there was a higher rate of device defects with SSL.
EXPERIMENTS: New graphene-philic surfactants carrying aromatic moieties in the hydrophilic headgroups and hydrophobic tails were synthesized by swapping the traditional sodium counterion with anilinium. 1H NMR spectroscopy was used to characterize the surfactants. These custom-made surfactants were used to assist the dispersion of GNPs in natural rubber latex matrices for the preparation of conductive nanocomposites. The properties of nanocomposites with the new anilinium surfactants were compared with commercial sodium surfactant sodium dodecylsulfate (SDS), sodium dodecylbenzenesulfonate (SDBS), and the previously synthesized aromatic tri-chain sodium surfactant TC3Ph3 (sodium 1,5-dioxo-1,5-bis(3-phenylpropoxy)-3-((3phenylpropoxy)carbonyl) pentane-2-sulfonate). Structural properties of the nanocomposites were studied using Raman spectroscopy, field emission scanning electron microscopy (FESEM), and high-resolution transmission electron microscopy (HRTEM). Electrical conductivity measurements and Zeta potential measurements were used to assess the relationships between total number of aromatic groups in the surfactant molecular structure and nanocomposite properties. The self-assembly structure of surfactants in aqueous systems and GNP dispersions was assessed using small-angle neutron scattering (SANS).
FINDINGS: Among these different surfactants, the anilinium version of TC3Ph3 namely TC3Ph3-AN (anilinium 1,5-dioxo-1,5-bis(3-phenylpropoxy)-3-((3phenylpropoxy)carbonyl) pentane-2-sulfonate) was shown to be highly efficient for dispersing GNPs in the NRL matrices, increasing electrical conductivity eleven orders of magnitude higher than the neat rubber latex. Comparisons between the sodium and anilinium surfactants show significant differences in the final properties of the nanocomposites. In general, the strategy of increasing the number of surfactant-borne aromatic groups by incorporating anilinium ions in surfactant headgroups appears to be effective.