Discovery of a first-in-class orally available HBV cccDNA inhibitor.

BACKGROUND & AIMS: The persistence of covalently closed circular DNA (cccDNA) in infected hepatocytes is the major barrier preventing viral eradication with existing therapies in patients with chronic hepatitis B. Therapeutic agents that can eliminate cccDNA are urgently needed to achieve viral eradication and thus HBV cure. METHODS: A phenotypic assay with HBV-infected primary human hepatocytes (PHHs) was employed to screen for novel cccDNA inhibitors. A HBVcircle mouse model and a uPA-SCID (urokinase-type plasminogen activator-severe combined immunodeficiency) humanized liver mouse model were used to evaluate the anti-HBV efficacy of the discovered cccDNA inhibitors. RESULTS: Potent and dose-dependent reductions in extracellular HBV DNA, HBsAg, and HBeAg levels were achieved upon the initiation of ccc_R08 treatment two days after the HBV infection of PHHs. More importantly, the level of cccDNA was specifically reduced by ccc_R08, while it did not obviously affect mitochondrial DNA. Additionally, ccc_R08 showed no significant cytotoxicity in PHHs or in multiple proliferating cell lines. The twice daily oral administration of ccc_R08 to HBVcircle model mice, which contained surrogate cccDNA molecules, significantly decreased the serum levels of HBV DNA and antigens, and these effects were sustained during the off-treatment follow-up period. Moreover, at the end of follow-up, the levels of surrogate cccDNA molecules in the livers of ccc_R08-treated HBVcircle mice were reduced to below the lower limit of quantification. CONCLUSIONS: We have discovered a small-molecule cccDNA inhibitor that reduces HBV cccDNA levels. cccDNA inhibitors potentially represent a new approach to completely cure patients chronically infected with HBV. IMPACT AND IMPLICATIONS: Covalently closed circular DNA (cccDNA) persistence in HBV-infected hepatocytes is the root cause of chronic hepatitis B. We discovered a novel small-molecule cccDNA inhibitor that can specifically reduce cccDNA levels in HBV-infected hepatocytes. This type of molecule could offer a new approach to completely cure patients chronically infected with HBV.

Toughening and Crack Healing Mechanisms in Nanotwinned Diamond Composites with Various Polytypes.

As an emerging ceramic material, recently synthesized nanotwinned diamond composites with various polytypes embedded in nanoscale twins exhibit unprecedented fracture toughness without sacrificing hardness. However, the toughening and crack healing mechanisms at the atomic scale and the associated crack propagation process of nanotwinned diamond composites remain mysterious. Here, we perform large-scale atomistic simulations of crack propagation in nanotwinned diamond composites to explore the underlying toughening and crack healing mechanisms in nanotwinned diamond composites. Our simulation results show that nanotwinned diamond composites have a higher fracture energy than single-crystalline and nanotwinned diamonds, which originates from multiple toughening mechanisms, including twin boundary and phase boundary impeding crack propagation, crack deflection and zigzag paths in nanotwins and sinuous paths in polytypes, and the formation of disordered atom clusters. More remarkably, our simulations reproduce more detailed crack propagation processes at the atomic scale, which is inaccessible by experiments. Moreover, our simulations reveal that crack healing occurs due to the rebonding of atoms on fracture surfaces during unloading and that the extent of crack healing is associated with whether the crack surfaces are clean. Our current study provides mechanistic insights into a fundamental understanding of toughening and crack healing mechanisms in nanotwinned diamond composites.

Microbial community analysis of membrane bioreactor incorporated with biofilm carriers and activated carbon for nitrification of urine.

The integration of powdered activated carbon and biofilm carriers in a membrane bioreactor (MBR) presents a promising approach to address the challenge of long hydraulic retention time (HRT) for nitrification of hydrolysed urine. This study investigated the effect of the incorporation in the MBR on microbial dynamics, focusing on dominant nitrifying bacteria. The results showed that significant shifts in microbial compositions were observed with the feed transition to full-strength urine and across different sludge growth forms. Remarkably, the nitrite-oxidizing bacteria Nitrospira were highly enriched in the suspended sludge. Simultaneously, ammonia-oxidizing bacteria, Nitrosococcaceae thrived in the attached biomass, showing a significant seven-fold increase in relative abundance compared to its suspended counterpart. Consequently, the incorporated MBR displayed 36% higher nitrification rate and 40% HRT reduction compared to the conventional MBR. This study provides valuable insights on the potential development of household or building scale on-site nutrient recovery from urine to fertiliser.

Room-temperature super-elongation in high-entropy alloy nanopillars.

Nanoscale small-volume metallic materials typically exhibit high strengths but often suffer from a lack of tensile ductility due to undesirable premature failure. Here, we report unusual room-temperature uniform elongation up to ~110% at a high flow stress of 0.6-1.0 GPa in single-crystalline <110>-oriented CoCrFeNi high-entropy alloy nanopillars with well-defined geometries. By combining high-resolution microscopy and large-scale atomistic simulations, we reveal that this ultrahigh uniform tensile ductility is attributed to spatial and synergistic coordination of deformation twinning and dislocation slip, which effectively promote deformation delocalization and delay necking failure. These joint and/or sequential activations of the underlying displacive deformation mechanisms originate from chemical compositional heterogeneities at the atomic level and resulting wide variations in generalized stacking fault energy and associated dislocation activities. Our work provides mechanistic insights into superplastic deformations of multiple-principal element alloys at the nanoscale and opens routes for designing nanodevices with high mechanical reliability.

Application of Benchtop NMR for Metabolomics Study Using Feces of Mice with DSS-Induced Colitis.

Nuclear magnetic resonance (NMR)-based metabolomics, which comprehensively measures metabolites in biological systems and investigates their response to various perturbations, is widely used in research to identify biomarkers and investigate the pathogenesis of underlying diseases. However, further applications of high-field superconducting NMR for medical purposes and field research are restricted by its high cost and low accessibility. In this study, we applied a low-field, benchtop NMR spectrometer (60 MHz) employing a permanent magnet to characterize the alterations in the metabolic profile of fecal extracts obtained from dextran sodium sulfate (DSS)-induced ulcerative colitis model mice and compared them with the data acquired from high-field NMR (800 MHz). Nineteen metabolites were assigned to the 60 MHz 1H NMR spectra. Non-targeted multivariate analysis successfully discriminated the DSS-induced group from the healthy control group and showed high comparability with high-field NMR. In addition, the concentration of acetate, identified as a metabolite with characteristic behavior, could be accurately quantified using a generalized Lorentzian curve fitting method based on the 60 MHz NMR spectra.

Stretchable polymer composites with ultrahigh piezoelectric performance.

Flexible piezoelectric materials capable of withstanding large deformation play key roles in flexible electronics. Ferroelectric ceramics with a high piezoelectric coefficient are inherently brittle, whereas polar polymers exhibit a low piezoelectric coefficient. Here we report a highly stretchable/compressible piezoelectric composite composed of ferroelectric ceramic skeleton, elastomer matrix and relaxor ferroelectric-based hybrid at the ceramic/matrix interface as dielectric transition layers, exhibiting a giant piezoelectric coefficient of 250 picometers per volt, high electromechanical coupling factor keff of 65%, ultralow acoustic impedance of 3MRyl and high cyclic stability under 50% compression strain. The superior flexibility and piezoelectric properties are attributed to the electric polarization and mechanical load transfer paths formed by the ceramic skeleton, and dielectric mismatch mitigation between ceramic fillers and elastomer matrix by the dielectric transition layer. The synergistic fusion of ultrahigh piezoelectric properties and superior flexibility in these polymer composites is expected to drive emerging applications in flexible smart electronics.

3D printing of inorganic nanomaterials by photochemically bonding colloidal nanocrystals.

3D printing of inorganic materials with nanoscale resolution offers a different materials processing pathway to explore devices with emergent functionalities. However, existing technologies typically involve photocurable resins that reduce material purity and degrade properties. We develop a general strategy for laser direct printing of inorganic nanomaterials, as exemplified by more than 10 semiconductors, metal oxides, metals, and their mixtures. Colloidal nanocrystals are used as building blocks and photochemically bonded through their native ligands. Without resins, this bonding process produces arbitrary three-dimensional (3D) structures with a large inorganic mass fraction (~90%) and high mechanical strength. The printed materials preserve the intrinsic properties of constituent nanocrystals and create structure-dictated functionalities, such as the broadband chiroptical responses with an anisotropic factor of ~0.24 for semiconducting cadmium chalcogenide nanohelical arrays.

Fabrication of Transparent and Flexible Digital Microfluidics Devices.

This study proposed a fabrication method for thin, film-based, transparent, and flexible digital microfluidic devices. A series of characterizations were also conducted with the fabricated digital microfluidic devices. For the device fabrication, the electrodes were patterned by laser ablation of 220 nm-thick indium tin oxide (ITO) layer on a 175 µm-thick polyethylene terephthalate (PET) substrate. The electrodes were insulated with a layer of 12 µm-thick polyethylene (PE) film as the dielectric layer, and finally, a surface treatment was conducted on PE film in order to enhance the hydrophobicity. The whole digital microfluidic device has a total thickness of less than 200 µm and is nearly transparent in the visible range. The droplet manipulation with the proposed digital microfluidic device was also achieved. In addition, a series of characterization studies were conducted as follows: the contact angles under different driving voltages, the leakage current density across the patterned electrodes, and the minimum driving voltage with different control algorithms and droplet volume were measured and discussed. The UV-VIS spectrum of the proposed digital microfluidic devices was also provided in order to verify the transparency of the fabricated device. Compared with conventional methods for the fabrication of digital microfluidic devices, which usually have opaque metal/carbon electrodes, the proposed transparent and flexible digital microfluidics could have significant advantages for the observation of the droplets on the digital microfluidic device, especially for colorimetric analysis using the digital microfluidic approach.

CuH-Catalyzed Enantioselective Reductive Coupling of 1,3-Dienes and Trifluoromethyl Ketoimines or α-Iminoacetates.

The intermolecular addition of allylic copper species generated from diene and copper hydride remains elusive. Herein copper hydride catalyzed asymmetric cross reductive coupling of conjugated dienes and ketoimines including trifluoromethyl ketoimines and α-iminoacetates was first achieved using chiral Ph-BPE as the ligand, providing rapid access to structurally and optically enriched homoallylic amines containing two vicinal stereogenic centers with up to 95% yield, 99% ee, and 11:1 diastereoselectivities.

Interfacially Locked Metal Aerogel Inside Porous Polymer Composite for Sensitive and Durable Flexible Piezoresistive Sensors.

Flexible pressure sensors play significant roles in wearable devices, electronic skins, and human-machine interface (HMI). However, it remains challenging to develop flexible piezoresistive sensors with outstanding comprehensive performances, especially with excellent long-term durability. Herein, a facile "interfacial locking strategy" has been developed to fabricate metal aerogel-based pressure sensors with excellent sensitivity and prominent stability. The strategy broke the bottleneck of the intrinsically poor mechanical properties of metal aerogels by grafting them on highly elastic melamine sponge with the help of a thin polydimethylsiloxane (PDMS) layer as the interface-reinforcing media. The hierarchically porous conductive structure of the ensemble offered the as-prepared flexible piezoresistive sensor with a sensitivity as high as 12 kPa-1 , a response time as fast as 85 ms, and a prominent durability over 23 000 compression cycles. The excellent comprehensive performance enables the successful application of the flexible piezoresistive sensor as two-dimensional (2D) array device as well as three-dimensional (3D) force-detecting device for real-time monitoring of HMI activities.

Impact of source-separation of urine on treatment capacity, process design, and capital expenditure of a decentralised wastewater treatment plant.

In this study, the impact of urine diversion on the treatment capacity, treatment process, and capital costs of a decentralised wastewater treatment plant (WWTP) was simulated using BioWin. The data for simulation including for economic analysis were obtained from a real decentralised WWTP at Sydney. Simulation was conducted for two alternative process design scenarios of a WWTP: membrane bioreactor (MBR) without denitrification and anaerobic MBR in place of aerobic MBR and compared to existing process design. The simulation shows that with about 75% urine diversion (through source separation), the treatment capacity of the existing WWTP can be doubled although above 40% urine diversion, the impact appears less rapid. When the urine diversion exceeds 75%, it was found that the anoxic tank for biological denitrification becomes redundant and the current wastewater treatment process could be replaced with a simpler and much less aeration intensive membrane bioreactor (MBR) producing similar effluent quality with a 24% reduction in capital expenditure (footprint) cost. Anaerobic MBR can be a potential alternative to aerobic MBR although pre-treatment becomes essential before reverse osmosis treatment for water reuse applications. Sensitivity analysis has revealed that by operating the bioreactor at higher mixed liquor suspended solids concentrations (9 g/L instead of 5 g/L) could help increase the WWTP treatment capacity by about 3.5 times at 75% urine diversion. Hence, urine diversion (until nitrogen-limiting conditions occur above 75% urine diversion) can increase the treatment capacity of an existing WWTP and reduce the capital expenses due to reduced plant footprint.

Glucose-free fructose production from Jerusalem artichoke using a recombinant inulinase-secreting Saccharomyces cerevisiae strain.

PURPOSE OF WORK: Using inulin (polyfructose) obtained from Jerusalen artichokes, we have produced fructose free of residual glucose using a recombinant inulinase-secreting strain of Saccharomyces cerevisiae in a one-step fermentation of Jerusalem artichoke tubers. For producing fructose from inulin, a recombinant inulinase-producing Saccharomyce cerevisiae strain was constructed with a deficiency in fructose uptake by disruption of two hexokinase genes hxk1 and hxk2. The inulinase gene introduced into S. cerevisiae was cloned from Kluyveromyces cicerisporus. Extracellular inulinase activity of the recombinant hxk-mutated S. cerevisiae strain reached 31 U ml(-1) after 96 h growth. When grown in a medium containing Jerusalem artichoke tubers as the sole component without any additives, the recombinant yeast accumulated fructose up to 9.2% (w/v) in the fermentation broth with only 0.1% (w/v) glucose left after 24 h.

Fertiliser recovery from source-separated urine via membrane bioreactor and heat localized solar evaporation.

Urine with its abundant macronutrients (N-P-K) is an ideal resource for the production of fertiliser. However, the odor and pathogens in the raw urine must be removed to meet the public acceptance of urine collection systems and to enable its safe reuse as a fertiliser. In this work, real urine was collected and treated through a pilot-scale gravity-driven membrane bioreactor (GDMBR) to remove the malodorous organics and to nitrify almost 50% of the ammonia into nitrate. The stablised urine was subsequently distilled via low-cost heat localized solar evaporation (HLSE) to produce a non-odorous solid fertiliser. The developed HLSE with a small footprint can attract bulk solution into a vertical insulated space and quickly heat it up to 68 °C within 1 h. The HLSE process had vapour flux at 1.3 kg m-2 h-1 as well as high solar to vapour conversion efficiency at 87%. Based on the EDX mapping and XRD analysis, the generated crystals are mainly NaNO3, NH4Cl, NaCl, NH4H2PO4 and K2HPO4, which are ideal nutrients for vegetation. In this study, the produced urine-derived fertilisers have a better performance on the growth of the leafy basil than the all-purpose commercial fertilisers. Generally, the GDMBR-HLSE is a promising cost-effective and green technology for nutrients recovery from urine.

Ultrahigh specific strength in a magnesium alloy strengthened by spinodal decomposition.

Strengthening of magnesium (Mg) is known to occur through dislocation accumulation, grain refinement, deformation twinning, and texture control or dislocation pinning by solute atoms or nano-sized precipitates. These modes generate yield strengths comparable to other engineering alloys such as certain grades of aluminum but below that of high-strength aluminum and titanium alloys and steels. Here, we report a spinodal strengthened ultralightweight Mg alloy with specific yield strengths surpassing almost every other engineering alloy. We provide compelling morphological, chemical, structural, and thermodynamic evidence for the spinodal decomposition and show that the lattice mismatch at the diffuse transition region between the spinodal zones and matrix is the dominating factor for enhancing yield strength in this class of alloy.

Enhanced expression of heterologous inulinase in Kluyveromyces lactis by disruption of hap1 gene.

Inulinase gene (Kcinu) derived from Kluyveromyces cicerisporus was expressed extracellularly in Kluyveromyces lactis using an episomal vector directed by Kcinu promoter. The influence of hap1 gene disruption on the expression of inulinase was studied. Inulinase activity in the supernatant of the recombinant Klhap1Delta strain was 391 U ml(-1) after cultured 120 h, which was 2.2-fold that of the wild type host. The relative inulinase mRNA level of the Klhap1Delta strain was 11.3-fold that of the wild type strain, and the expression plasmid was more stable in the mutant host. Based on these results, the disruption of hap1 facilitated the high and stable expression of inulinase controlled by Kcinu promoter in K. lactis.

Urine Treatment on the International Space Station: Current Practice and Novel Approaches.

A reliable, robust, and resilient water recovery system is of paramount importance on board the International Space Station (ISS). Such a system must be able to treat all sources of water, thereby reducing resupply costs and allowing for longer-term space missions. As such, technologies able to dewater urine in microgravity have been investigated by different space agencies. However, despite over 50 years of research and advancements on water extraction from human urine, the Urine Processing Assembly (UPA) and the Water Processor Assembly (WPA) now operating on the ISS still achieve suboptimal water recovery rates and require periodic consumables resupply. Additionally, urine brine from the treatment is collected for disposal and not yet reused. These factors, combined with the need for a life support system capable of tolerating even dormant periods of up to one year, make the research in this field ever more critical. As such, in the last decade, extensive research was conducted on the adaptation of existing or emerging technologies for the ISS context. In virtue of having a strong chemical resistance, small footprint, tuneable selectivity and versatility, novel membrane-based processes have been in focus for treating human urine. Their hybridisation with thermal and biological processes as well as the combination with new nanomaterials have been particularly investigated. This article critically reviews the UPA and WPA processes currently in operation on the ISS, summarising the research directions and needs, highlighted by major space agencies, necessary for allowing life support for missions outside the Low Earth Orbit (LEO). Additionally, it reviews the technologies recently proposed to improve the performance of the system as well as new concepts to allow for the valorisation of the nutrients in urine or the brine after urine dewatering.

Bromide and iodide selectivity in membrane capacitive deionisation, and its potential application to reduce the formation of disinfection by-products in water treatment.

The formation of toxic disinfection by-products during water disinfection due to the presence of bromide and iodide is a major concern. Current treatment technologies such as membrane, adsorption and electrochemical processes have been known to have limitations such as high energy demand and excessive chemical use. In this study, the selectivity between bromide and iodide, and their removal in membrane capacitive deionisation (MCDI) was evaluated. The results showed that iodide was more selectively removed over bromide from several binary feed waters containing bromide and iodide under various initial concentrations and applied voltages. Even in the presence of significant background concentration of sodium chloride, definite selectivity of iodide over bromide was observed. The high partial-charge transfer coefficient of iodide compared to bromide could be a feasible explanation for high iodide selectivity since both bromide and iodide have similar ionic charge and hydrated radius. The result also shows that MCDI can be a potential alternative for the removal of bromide and iodide during water treatment.

Venous arterialization for the treatment of large-area foot skin retrograde avulsion.

Between 2009 and 2011, three patients with large-area foot skin retrograde avulsion (more than 1% of the body surface area) underwent venous arterialization. Anastomosis of the artery in the wound surface with the vein in the skin flap and an appropriate number of venous end-to-end anastomoses were performed. The skin flaps survived in all 3 patients. Six months postoperatively, the flap elasticity and appearance were close to that of normal skin, and foot function was better without scar contracture. When venous arterialization is used to treat foot avulsion, the following points should be noted. Surgical indications include no fresh bleeding from the wound edge of the avulsed skin after debridement, more complete avulsed skin, and superficial veins that do not completely separate from the avulsed skin. Venous arterialization is not suitable to avulsion with fresh bleeding, avulsed skin in small fragments, and avulsion with a subcutaneous venous network embolism. During debridement, the subcutaneous venous network should be protected to avoid exposing the vein stems outside the fat layer. If the avulsion is less than 1% of the body surface area, arterial-venous anastomosis can provide adequate blood supply. Venous-venous anastomosis is performed as much as possible to enhance venous return and decrease microcirculatory pressure, which is conducive to the establishment of effective blood circulation.

[Simultaneous saccharification and fermentation of Jerusalem artichoke tubers to ethanol with an inulinase-hyperproducing yeast Kluyveromyces cicerisporus].

Jerusalem artichoke tubers with inulin as major component are potential feedstock for fuel ethanol production, and Kluyveromyces cicerisporus Y179 expressing high level of inulinase is suitable for ethanol production with this feedstock by simultaneous saccharification and fermentation approach. In this article, the impact of inoculum, aeration and temperature on ethanol production by the yeast was studied. The experimental results illustrated that inoculum with different levels and seed collected at different cultivation times had negligible effect, while anaerobic conditions enhanced ethanol production, and more ethanol was produced by the yeast at 30 degrees C than at 37 degrees C or 42 degrees C. The medium using Jerusalem artichoke tuber meal as sole component with 22% (W/V) total sugars was inoculated with 36 h-precultured seed at 10% (V/V), and the batch fermentation was conducted in a 5 L fermentor at 30 degrees C with a stirring speed of 300 r/min under anaerobic conditions. After 144 h, 12.3% (V/V) ethanol was produced and the yield of ethanol from sugars was 86.9% of its theoretical one, with 93.6% sugars consumed. These results indicate that K. cicerisporus Y179 is a promising candidate for industrial ethanol production using Jerusalem artichoke tuber feedstock.