Manufacture of Multilayered Artificial Cell Membranes through Sequential Bilayer Deposition on Emulsion Templates

Efforts to manufacture artificial cells that replicate the architectures, processes and behaviours of biological cells are rapidly increasing. Perhaps the most commonly reconstructed cellular structure is the membrane, through the use of unilamellar vesicles as models. However, many cellular membranes, including bacterial double membranes, nuclear envelopes, and organelle membranes, are multilamellar. Due to a lack of technologies available for their controlled construction, multilayered membranes are not part of the repertoire of cell-mimetic motifs used in bottom-up synthetic biology. To address this, we developed emulsion-based technologies that allow cell-sized multilayered vesicles to be produced layer-by-layer, with compositional control over each layer, thus enabling studies that would otherwise remain inaccessible. We discovered that bending rigidities scale with the number of layers and demonstrate inter-bilayer registration between coexisting liquid–liquid domains. These technologies will contribute to the exploitation of multilayered membrane structures, paving the way for incorporating protein complexes that span multiple bilayers.     

Semi-volatile organic compounds in the Campaspe River system (Victoria, Australia)

The levels of semi-volatile organic compounds (SVOCs) in the Campaspe River system, North Central Victoria, Australia, were monitored from January to September 1995, and the environmental trends and sources of these compounds were determined. The land use within particular catchments dictated the complexity of the SVOCs in run-off water. Run-off from native forest contained the greatest number of SVOCs. Soil and ground litter were shown to contain the greatest complexity of SVOCs, mostly terpenes. However, only a small proportion were leached by run-off with the remainder being tightly bound to the soil or volatilizing to the atmosphere. Under the prevailing drought of early 1995, the rivers contained no detectable SVOCs until the first catchment run-off after significant rainfall in May. The maximum complexity and concentration of the SVOCs coincided with extensive run-off after heavy rainfall in June, effecting a surge of SVOCs with the flood peak. Fatty acids, their amides and hydrocarbons made up the majority of the compounds identified and were in the 1-39 micrograms/l concentration range. The levels then diminished as the catchment was flushed by continual run-off. These results support Lake's 1967 hypothesis that natural floodwaters contain a signature of SVOCs entering from the catchment. The downstream passage of these compounds was obstructed by a man-made storage, Lake Eppalock, with only low concentrations of SVOCs (1-3 micrograms/l) in the outflow, displaying no correlation to run-off.     

Sourcing faecal pollution: a combination of library-dependent and library-independent methods to identify human faecal pollution in non-sewered catchments

Library-dependent (LD) (biochemical fingerprinting of Escherichia coli and enterococci) and library-independent (LI) (PCR detection of human-specific biomarkers) methods were used to detect human faecal pollution in three non-sewered catchments. In all, 550 E. coli isolates and 700 enterococci isolates were biochemically fingerprinted from 18 water samples and compared with metabolic fingerprint libraries of 4508 E. coli and 4833 enterococci isolates. E. coli fingerprints identified human unique biochemical phenotypes (BPTs) in nine out of 18 water samples; similarly, enterococci fingerprints identified human faecal pollution in 10 water samples. Seven samples were tested by PCR for the detection of biomarkers. Human-specific HF134 Bacteroides and enterococci surface protein (esp) biomarkers were detected in five samples. Four samples were also positive for HF183 Bacteroides biomarker. The combination of biomarkers detected human faecal pollution in six out of seven water samples. Of the seven samples analysed for both the indicators/markers, at least one indicator/marker was detected in every sample. Four of the seven PCR-positive samples were also positive for one of the human-specific E. coli or enterococci BPTs. The results indicated human faecal pollution in the studied sub-catchments after storm events. LD and LI methods used in this study complimented each other and provided additional information regarding the polluting sources when one method failed to detect human faecal pollution. Therefore, it is recommended that a combination of methods should be used to identify the source(s) of faecal pollution where possible.     

Binding Mode and Structure–Activity Relationships around Direct Inhibitors of the Nrf2–Keap1 Complex

An X‐ray crystal structure of Kelch‐like ECH‐associated protein (Keap1) co‐crystallised with (1S,2R)‐2‐[(1S)‐1‐[(1,3‐dioxo‐2,3‐dihydro‐1H‐isoindol‐2‐yl)methyl]‐1,2,3,4‐tetrahydroisoquinolin‐2‐carbonyl]cyclohexane‐1‐carboxylic acid (compound (S,R,S)‐ 1 a ) was obtained. This X‐ray crystal structure provides breakthrough experimental evidence for the true binding mode of the hit compound (S,R,S)‐ 1 a , as the ligand orientation was found to differ from that of the initial docking model, which was available at the start of the project. Crystallographic elucidation of this binding mode helped to focus and drive the drug design process more effectively and efficiently.     

Electrically Enhanced Boron Removal from Silicon Using Slag

An approach to enhance silicon refining using slag has been developed. The enhancement of the process was carried out by applying electrical potential difference across the slag and the silicon phase. This resulted in a shift in the apparent equilibrium in favor of higher partition ratio for the impurities. The application of electrical potential difference also enhanced the mass transfer rate increasing the overall kinetics of the process. This has been demonstrated in laboratory experimentations for the removal of boron from silicon-boron melts using slag. A CaO-SiO₂-Al₂O₃ slag was reacted with Si-B melt at 1823 K (1550 °C). Electrical potential differences were applied through graphite rods immersed in each of the liquid phase. The results showed that the apparent B-partition ratio and the apparent slag mass transfer coefficient were increased by a factor of 1.2 and 1.4, respectively, when a potential difference of 3 V was applied to the phases. The technique has the potential to be used for improving the existing slag refining process by increasing the overall kinetics and the slag capacity to absorb the impurities.     

Consumer Assessment of Beef Tenderloin Steaks from Various USDA Quality Grades at 3 Degrees of Doneness

A consumer study was conducted to determine palatability ratings of beef tenderloin steaks from USDA Choice, USDA Select, and USDA Select with marbling scores from Slight 50 to 100 (USDA High Select) cooked to various degrees of doneness. Steaks were randomly assigned to 1 of 3 degree of doneness categories: very‐rare, medium‐rare, or well‐done. Consumers (N = 315) were screened for preference of degree of doneness and fed 4 samples of their preferred doneness (a warm‐up and one from each USDA quality grade treatment in a random order). Consumers evaluated steaks on an 8‐point verbally anchored hedonic scale for tenderness, juiciness, flavor, and overall like as well as rated steaks as acceptable or unacceptable for all palatability traits. Quality grade had no effect (P > 0.05) on consumer ratings for tenderness, juiciness, flavor, and overall like scores, with all traits averaging above a 7 (“like very much”) on the 8‐point scale. In addition, no differences (P > 0.05) were found in the percentage of samples rated as acceptable for all palatability traits, with more than 94% of samples rated acceptable for each trait in all quality grades evaluated. Steaks cooked to well‐done had lower (P < 0.05) juiciness scores than steaks cooked to very‐rare or medium‐rare and were rated lower for tenderness (P < 0.05) than steaks cooked to a very‐rare degree of doneness. Results indicate consumers were not able to detect differences in tenderness, juiciness, flavor, or overall like among beef tenderloin steaks from USDA Choice and Select quality grades.     

Analysis of steelmaking kinetics from IMPHOS pilot plant data

It is widely accepted that understanding the kinetics of steelmaking is a complex task, and reliable and validated kinetics models are required for developing successful steelmaking process models. Therefore, as an initial attempt, this paper analyses the applicability of first order kinetics to explain the steelmaking reaction kinetics using the published data in the IMproving Phosphorus Refining research report. The process data for 20 heats in a 6?tonne pilot plant were analysed for the removal of carbon, silicon, manganese and phosphorus using first order kinetics with static and dynamic equilibrium conditions. It was observed that the removal behaviour of silicon closely followed a first order kinetics relationship, while that of carbon only approximately followed a first order kinetics relationship. The removal of manganese did not show a good degree of fit with first order kinetics using static equilibrium condition, but a clear improvement was observed when calculated using dynamic equilibrium condition. In contrast, the kinetics of phosphorus oxidation did not follow any first order relationship.     

Mechanical properties of morcellised bone graft with the addition of hydroxyapatite

Mixtures of morcellised bone graft (MBG) and hydroxyapatite (HA) are frequently used in revision arthroplasty surgery. However, the changes in the mechanical properties from adding HA to MBG are unknown. This study used a uniaxial compression test to replicate impaction bone grafting and subsequent early postoperative weightbearing to investigate the effect of adding different proportion of HA to MBG. To achieve this aim, human MBG was subjected to increasing impaction forces and the apparent stiffness and creep for each stress level determined. Subsequently, increasing proportions porous and non porous HA were added to the MBG. The major findings were that the apparent stiffness for MBG increased and the associated creep decreased both with the application of increasing stress and with the addition of increasing proportions of HA. In conclusion, greater proportions of HA in the graft mixture improved the mechanical response compared with MBG impacted under the same force. This improvement replicated the properties of pure MBG under high axial stress. This study indicates that graft mixtures of MBG and HA can be tailormade for patients. The need for less impaction force in MBG:HA mixtures to obtain the same properties as pure MBG may decrease the risk of intraoperative fracture.