From supercritical carbon dioxide to gas expanded liquids in extraction and chromatography of lipids

Analytical‐scale extraction and chromatography of oils, fats and other liposoluble compounds can be achieved by using supercritical carbon dioxide as a solvent. Since the 90's when supercritical fluid chromatography (SFC) was a hot topic, this technology has developed into a robust, modern analytical technique that uses any proportions of compressed CO₂ mixed with an organic solvent. Supercritical fluid extraction (SFE) on the other hand is only recently starting to reform in a similar way, towards the use of more robust extraction system and enabling mixing of compressed CO₂ with larger proportions of organic solvents. In this Feature article, the development of SFC and SFE into what options we have today is described, including the latest trend of using CO₂‐expanded liquid (CXL) as extraction solvent for lipids.     

A Model Framework for Transportation and Community Plan Analysis

A major weakness of present planning models is their neglect of the distributional consequences that result from alternative transportation and urban development plans. This shortcoming has serious implications for the evaluation and plan selection process because the costs and benefits considered can, by definition, be no more than partial. The model framework presented here makes explicit certain of these consequences; in particular, changes in the level of transportation service available to specific groups in the community, However, the approach described in this article does not represent a comprehensive framework for plan analysis and evaluation. For example, it does not consider the externalities generated by transportation plans, such as the social and environmental impacts resulting from the physical presence of transportation facilities. The model does, however, have the advantage of analyzing certain of the service characteristics of the transportation system at the micro scale, where they are more compatible with the treatment of externalities. Both service characteristics and externalities can then be incorporated in a comprehensive evaluation framework, such as those suggested by Hill (1968), Lichfield (1969), and, more recently Turner (1972).     

An Engineered Cholesterol Oxidase Catalyses Enantioselective Oxidation of Non-steroidal Secondary Alcohols

The enantioselective oxidation of 2° alcohols to ketones is an important reaction in synthetic chemistry, especially if it can be achieved using O₂-driven alcohol oxidases under mild reaction conditions. However to date, oxidation of secondary alcohols using alcohol oxidases has focused on activated benzylic or allylic substrates, with unactivated secondary alcohols showing poor activity. Here we show that cholesterol oxidase (EC 1.1.3.6) could be engineered for activity towards a range of aliphatic, cyclic, acyclic, allylic and benzylic secondary alcohols. Additionally, since the variants demonstrated high (S)-selectivity, deracemisation reactions were performed in the presence of ammonia borane to obtain enantiopure (R)-alcohols.     

Clinical and Genetic Re-Evaluation of Inherited Retinal Degeneration Pedigrees following Initial Negative Findings on Panel-Based Next Generation Sequencing

Although rare, inherited retinal degenerations (IRDs) are the most common reason for blind registration in the working age population. They are highly genetically heterogeneous (>300 known genetic loci), and confirmation of a molecular diagnosis is a prerequisite for many therapeutic clinical trials and approved treatments. First-tier genetic testing of IRDs with panel-based next-generation sequencing (pNGS) has a diagnostic yield of ≈70–80%, leaving the remaining more challenging cases to be resolved by second-tier testing methods. This study describes the phenotypic reassessment of patients with a negative result from first-tier pNGS and the rationale, outcomes, and cost of second-tier genetic testing approaches. Removing non-IRD cases from consideration and utilizing case-appropriate second-tier genetic testing techniques, we genetically resolved 56% of previously unresolved pedigrees, bringing the overall resolve rate to 92% (388/423). At present, pNGS remains the most cost-effective first-tier approach for the molecular assessment of diverse IRD populations Second-tier genetic testing should be guided by clinical (i.e., reassessment, multimodal imaging, electrophysiology), and genetic (i.e., single alleles in autosomal recessive disease) indications to achieve a genetic diagnosis in the most cost-effective manner.     

Hot-rolling and forging of mica-filled polyolefins

As in the case of fibers, the mechanical properties of plastic composites containing mica flakes are extremely sensitive to flake orientation in the direction of an applied stress, so that even a small angular displacement can cause major reductions in the strength, modulus, and fracture toughness. In order to encourage parallel alignment of mica flakes in a thermoplastic composite, two methods of flow orientation were examined. In the first series, rectangular billets of mica-filled, high-density polyethylene were hot-pressed in order to cause longitudinal melt flow in a narrow channel. A parallel series of experiments was also carried out with mica-filled polypropylene in which the composite was extruded and calendered into a thin, continuous strip. In both processing techniques, the resulting extensional flow produced large increases in the tensile and flexural properties. The performance of the mica-filled poly-propylene was limited by its tendency to fibrillate during rolling. Drop-impact measurements recorded a four-fold increase in fracture toughness. The increased tensile and flexural properties were attributed to both the greater degree of parallel alignment of the mica flakes and the increased molecular orientation in the direction of flow. Such flow orientation methods appear necessary if the full benefit of mica-flake reinforcement is to be achieved.     

Enhanced efficacy and sensory properties of an anti‐dandruff shampoo containing zinc pyrithione and climbazole

Dandruff is a common complaint and is suffered by as much as half of the population at some time post puberty. The condition is characterized by the presence of flakes on the scalp and in the hair, and is often accompanied by itch. The most common treatment for dandruff is the use of shampoo formulations that contain fungistatic agents such as zinc pyrithione (ZPT) and octopirox. Whilst most antidandruff shampoos are effective in resolving the symptoms of dandruff these shampoos can often result in hair condition that is less than acceptable to consumers which can lead to a tendency for them to revert to use of a non‐antidandruff shampoo. This can result in a rapid return of dandruff symptoms. The aim of this investigation was to study the impact of using a combination of antidandruff actives and silicones on the resolution of dandruff and to deliver superior sensory properties to the hair. We have demonstrated that shampoo containing the dual active system of ZPT/Climbazole deposits both active agents onto a model skin surface (VitroSkin) and reduces Malassezia furfur regrowth in vitro. Clinical evaluation of the dual active shampoo demonstrated superior efficacy and retained superiority during a regression phase where all subjects reverted to using a non‐antidandruff shampoo. We have also demonstrated that it is possible to deposit silicone materials from antidandruff shampoo uniformly over both virgin and damaged hair fibres that results in smoother hair fibres (as evidenced by reduced dry friction). This combination of antidandruff agents and conditioning silicones delivered from a shampoo provides subjects with superior antidandruff efficacy and desired end sensory benefits ensuring compliance and longer term dandruff removal.     

Characterisation of CYP102A25 from Bacillus marmarensis and CYP102A26 from Pontibacillus halophilus: P450 Homologues of BM3 with Preference towards Hydroxylation of Medium‐Chain Fatty Acids

Cytochrome P450 monooxygenases are highly desired biocatalysts owing to their ability to catalyse a wide variety of chemically challenging C?H activation reactions. The CYP102A subfamily of enzymes are natural catalytically self‐sufficient proteins consisting of a haem and FMN‐FAD reductase domain fused in a single‐component system. They catalyse the oxygenation of saturated and unsaturated fatty acids to produce primarily ω?1, ω?2 and ω?3 hydroxy acids. These monooxygenases have potential applications in biotechnology; however, their substrate range is still limited and there is a continued need to add diversity to this class of biocatalysts. Herein, we present the characterisation of two new members of this class of enzymes, CYP102A25 (BMar) from Bacillus marmarensis and CYP102A26 (PHal) from Pontibacillus halophilus, both of which express readily in a recombinant bacterial host. BMar exhibits the highest activity toward myristic acid and shows moderate activity towards unsaturated fatty acids. PHal exhibits broader activity towards mid‐chain‐saturated (C₁₄–C₁₈) and unsaturated fatty acids. Furthermore, PHal shows good regioselectivity for the hydroxylation of myristic acid, targeting the ω?2 position for C?H activation.     

Fish intake and serum levels of organochlorines among Japanese women

This study evaluates background serum levels of selected organochlorine compounds among Japanese women of reproductive age and investigates whether lifestyle factors, especially dietary factors, may be associated with these levels. A cross-sectional study was performed on 80 Japanese women, aged 26-43 years, who complained of infertility and were confirmed not to have endometriosis. The serum levels of total toxic equivalency (TEQ), 18 polychlorinated dibenzo-p-dioxins (PCDDs)/polychlorinated dibenzofurans (PCDFs), 4 coplanar polychlorinated biphenyls (cPCBs), 36 ortho-substituted polychlorinated biphenyls (PCBs), and 13 chlorinated pesticides or their metabolites were measured and data were collected on the women's age, residence, occupation, body mass index (BMI), smoking and alcohol habit and 6 dietary intakes (fish, meats, rice, vegetables, fruits and dairy products). The serum median level of total TEQ was 25.1 pg TEQ/g lipid, that of PCDDs/PCDFs/cPCBs was 11.5 pmol/g lipid, that of PCBs was 0.46 nmol/g lipid, and that of total pesticides was 1.32 nmol/g lipid. The serum levels of total TEQ, PCDDs/PCDFs/cPCBs, PCBs and pesticides were positively associated with age (P for trend=0.003, 0.01, 0.005 and 0.01, respectively) and frequent fish consumption (P for trend=0.002, 0.003, 0.0003 and 0.006, respectively). Other lifestyle factors were not associated with serum organochlorine levels. The present study suggests that Japanese women who consume fish frequently in their reproductive period tend to accumulate organochlorines in their bodies.     

Investigating the suitability of electrohydrodynamic lithography for the fabrication of cell substrates

We have investigated the feasibility of using electrohydrodynamic lithography (EHDL) to produce large lateral areas of patterned polymer substrates for use in biomedical applications. Half centimetered squared regions of uninterrupted patterning were obtained and characterised by profilometry and microscopy. The patterning was found to be elliptical in shape and was composed of concentric bands of distinct patterns centred on initial, randomly located nucleation sites. The size of the patterned area was limited by the degree to which the inter-electrode gap could be kept perfectly parallel, with a difference in height of just 15 nm influencing the patterning. Such sensitivity meant that issues such as the stiffness of the electrodes were important. Hierarchical patterning was achieved by combining EHDL with fracture-induced structuring. The substrates fabricated using EHDL were demonstrated to be viable for cell culture in vitro using fibroblast (3T3) and muscle (C2C12) cell lines.