Malic acid production from renewables: a review

Malic acid derived from fossil resources is currently applied in the food and beverage industries with a medium global production capacity. However, in the transition from a fossil-based to a bio-based economy, biotechnologically produced l -malic acid may become an important platform chemical with many new applications, especially in the field of biopolymers. In this review, currently used petrochemical production routes to dl -malic acid are outlined and insights into possible bio-based alternatives for microbial l -malic acid production are provided. Besides ecological reasons, the possibility to produce enantiopure l -malic acid by microbial fermentation is the biggest advantage over chemical synthesis. State-of-the-art and open challenges concerning production host engineering, substrate choice and downstream processing are addressed. With regard to production hosts, a literature overview is given covering the leading natural production strains of Aspergillus, Ustilago and Aureobasidium, as well as Escherichia coli as the most important engineered recombinant host. The utilization of renewable substrates as an alternative to glucose is emphasized in particular as a key aspect for a competitive bio-based production. Out of the alternative substrates discussed in this review, the industrial side-streams crude glycerol and molasses seem to be most promising for large-scale l -malic acid production. © 2019 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Vegetation Change in Great Lakes Coastal Wetlands: Deviation from the Historical Cycle

Water-level change is integral to the structure and function of Great Lakes coastal wetlands, and many studies document predictable relationships between vegetation and water level. However, anthropogenic stressors, such as invasive species, land-use change, and water-level stabilization, interact to shift the historical cycle (of native vegetation migration up- and down-slope) toward dominance by invasive Typha species. Knowing from earlier studies that water-level stabilization alters the historical vegetation cycle, we asked if similar shifts can occur where water levels are not stabilized. Using historical aerial photographs of three coastal wetlands (in Lake Michigan's Green Bay, Wisconsin), we determined that habitat dominated by Typha species has expanded to eliminate wet meadow habitat. Between 1974 and 1992, linear regressions showed strong, significant relationships of both meadow area (R² ≥ 0.894; p < 0.02) and marsh area (R² ≥ 0.784; p < 0.05) to water level in all three wetlands. In 2000, meadow area was below that predicted by the historical pattern due to the landward advance of marsh habitat during a year of decreasing water levels. In the same period, land use in the wetland watersheds converted from agriculture to urban. Urbanization and the replacement of native Typha latifolia by the invasive hybrid Typha xglauca may have overwhelmed the beneficial impact of water-level fluctuation. The documentation of vegetation shifts, as herein, is an essential step in the process of preserving and restoring ecological integrity.     

Synthesis and Pharmacological In Vitro Investigations of Novel Shikonin Derivatives with a Special Focus on Cyclopropane Bearing Derivatives

Melanoma is the deadliest form of skin cancer and accounts for about three quarters of all skin cancer deaths. Especially at an advanced stage, its treatment is challenging, and survival rates are very low. In previous studies, we showed that the constituents of the roots of Onosma paniculata as well as a synthetic derivative of the most active constituent showed promising results in metastatic melanoma cell lines. In the current study, we address the question whether we can generate further derivatives with optimized activity by synthesis. Therefore, we prepared 31, mainly novel shikonin derivatives and screened them in different melanoma cell lines (WM9, WM164, and MUG-Mel2 cells) using the XTT viability assay. We identified (R)-1-(1,4-dihydro-5,8-dihydroxy-1,4-dioxonaphthalen-2-yl)-4-methylpent-3-enyl 2-cyclopropyl-2-oxoacetate as a novel derivative with even higher activity. Furthermore, pharmacological investigations including the ApoToxGlo^TM Triplex assay, LDH assay, and cell cycle measurements revealed that this compound induced apoptosis and reduced cells in the G1 phase accompanied by an increase of cells in the G2/M phase. Moreover, it showed hardly any effects on the cell membrane integrity. However, it also exhibited cytotoxicity against non-tumorigenic cells. Nevertheless, in summary, we could show that shikonin derivatives might be promising drug leads in the treatment of melanoma.     

Structure-Activity Relationships of Benzamides and Isoindolines Designed as SARS-CoV Protease Inhibitors Effective against SARS-CoV-2

Inhibition of coronavirus (CoV)-encoded papain-like cysteine proteases (PL^pro) represents an attractive strategy to treat infections by these important human pathogens. Herein we report on structure-activity relationships (SAR) of the noncovalent active-site directed inhibitor (R)-5-amino-2-methyl-N-(1-(naphthalen-1-yl)ethyl) benzamide ( 2 b ), which is known to bind into the S3 and S4 pockets of the SARS-CoV PL^pro. Moreover, we report the discovery of isoindolines as a new class of potent PL^pro inhibitors. The studies also provide a deeper understanding of the binding modes of this inhibitor class. Importantly, the inhibitors were also confirmed to inhibit SARS-CoV-2 replication in cell culture suggesting that, due to the high structural similarities of the target proteases, inhibitors identified against SARS-CoV PL^pro are valuable starting points for the development of new pan-coronaviral inhibitors.     

Characterization and tests of planar Co3O4 model catalysts prepared by chemical vapor deposition

The chemical vapor deposition method was used to deposit thin films of cobalt oxide starting with cobalt (II) acetylacetonate and oxygen. The deposition process was investigated and the obtained films were identified as a cubic spinel-type polycrystalline Co₃O₄ with a crystallite size of 30–40 nm. The coating was carbon-free and the surface oxygen concentration was measured to be 66 at.% with AES analysis. Smooth and highly uniform thin films were deposited on planar stainless steel substrates and subjected to TPR and catalysis tests that show positive correlation. The apparent activation energy of Co₃O₄ reduction to CoO was measured to be (33±5) kJ/mol. The catalytic activity of Co₃O₄ was investigated toward the conversion of both propane and ethanol to carbon dioxide. Though the catalytic action was registered at the same temperature, the deactivation process was seen to be different. The catalytic conversion of ethanol induces a fast deactivation process, which was linked to its high ability to reduce Co₃O₄.     

Effect of particle dimension on biocompatibility of carbon nanomaterials

With various emerging applications ranging from medicine to materials and electronics, the risk of exposure to nanomaterials is rapidly increasing. Several routes of exposure to nanomaterials exist; the most important being dermal contact and inhalation. In this dermal toxicity study, the cellular effects of carbon-based materials with diameters ranging from micro- to nano-dimension were investigated using mouse keratinocytes (HEL-30). The carbon materials tested included carbon fibers (CF; 10 μm diameter), carbon nanofibers (CNF; 100 nm diameter), multi-walled carbon nanotubes (MWCNT; 10 nm diameter), and single-walled carbon nanotubes (SWCNT; 1 nm diameter). CF and CNF did not significantly affect cell viability; however, MWCNT and SWCNT reduced cell viability in a time-dependent manner up to 48 h, with full recovery of mitochondrial function by the 72 h time point. After a 24 h exposure, cells exposed to MWCNT produced up to 3-fold higher increase in reactive oxygen species than those exposed to SWCNT. The results of this study suggest that high-aspect ratio carbon material toxicity is dependent on dimension and composition.     

Attention: Reaction Time and Accuracy Reveal Different Mechanisms.

The authors propose that there are 2 different mechanisms whereby spatial cues capture attention. The voluntary mechanism is the strategic allocation of perceptual resources to the location most likely to contain the target. The involuntary mechanism is a reflexive orienting response that occurs even when the spatial cue does not indicate the probable target location. Voluntary attention enhances the perceptual representation of the stimulus in the cued location relative to other locations. Hence, voluntary attention affects performance in experiments designed around both accuracy and reaction time. Involuntary attention affects a decision as to which location should be responded to. Because involuntary attention does not change the perceptual representation, it affects performance in reaction time experiments but not accuracy experiments. The authors obtained this pattern of results in 4 different versions of the spatial cuing paradigm. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

SK119, a Novel Shikonin Derivative,Leads to Apoptosis in Melanoma Cell Lines and Exhibits Synergistic Effects with Vemurafenib and Cobimetinib

Melanoma is a complex and heterogenous disease, displays the deadliest form of skin cancer, and accounts for approx. 80% of all skin cancer deaths. In this study, we reported on the synthesis and pharmacological effects of a novel shikonin derivative (SK119), which is active in a nano-molar range and exhibits several promising in vitro effects in different human melanoma cells. SK119 was synthesized from shikonin as part of our search for novel, promising shikonin derivatives. It was screened against a panel of melanoma and non-tumorigenic cell lines using XTT viability assays. Moreover, we studied its pharmacological effects using apoptosis and Western blot experiments. Finally, it was combined with current clinically used melanoma therapeutics. SK119 exhibited IC₅₀ values in a nano-molar range, induced apoptosis and led to a dose-dependent increase in the expression and protein phosphorylation of HSP27 and HSP90 in WM9 and MUG-Mel 2 cells. Combinatorial treatment, which is highly recommended in melanoma, revealed the synergistic effects of SK119 with vemurafenib and cobimetinib. SK119 treatment changed the expression levels of apoptosis genes and death receptor expression and exhibited synergistic effects with vemurafenib and cobimetinib in human melanoma cells. Further research indicates a promising potential in melanoma therapy.     

A case study on software risk analysis and planning in medical device development

Software failures in medical devices can lead to catastrophic situations. Therefore, it is crucial to handle software-related risks when developing medical devices, and there is a need for further analysis of how this type of risk management should be conducted. The objective of this paper is to collect and summarise experiences from conducting risk management with an organisation developing medical devices. Specific focus is put on the first steps of the risk management process, i.e. risk identification, risk analysis, and risk planning. The research is conducted as action research, with the aim of analysing and giving input to the organisation’s introduction of a software risk management process. First, the method was defined based on already available methods and then used. The defined method focuses on user risks, based on scenarios describing the expected use of the medical device in its target environment. During the use of the method, different stakeholders, including intended users, were involved. Results from the case study show that there are challenging problems in the risk management process with respect to definition of the system boundary and system context, the use of scenarios as input to the risk identification, estimation of detectability during risk analysis, and action proposals during risk planning. It can be concluded that the risk management method has potential to be used in the development organisation, although future research is needed with respect to, for example, context limitation and how to allow for flexible updates of the product.