The increased cultivation of highly productive C4 crop plants may contribute to a second green revolution in agriculture. However, the regulation of mineral nutrition is rather poorly understood in C4 plants. To understand the impact of C4 photosynthesis on the regulation of sulfate uptake by the root and sulfate assimilation into cysteine at the whole plant level, seedlings of the monocot C4 plant maize (Zea mays) were exposed to a non-toxic level of 1.0 µl l−1 atmospheric H2S at sulfate-sufficient and sulfate-deprived conditions. Sulfate deprivation not only affected growth and the levels of sulfur- and nitrogen-containing compounds, but it also enhanced the expression and activity of the sulfate transporters in the root and the expression and activity of APS reductase (APR) in the root and shoot. H2S exposure alleviated the establishment of sulfur deprivation symptoms and seedlings switched, at least partly, from sulfate to H2S as sulfur source. Moreover, H2S exposure resulted in a downregulation of the expression and activity of APR in both shoot and root, though it hardly affected that of the sulfate transporters in the root. These results indicate that maize seedlings respond similarly to sulfate deprivation and atmospheric H2S exposure as C3 monocots, implying that C4 photosynthesis in maize is not associated with a distinct whole plant regulation of sulfate uptake and assimilation into cysteine.
Hyperhomocysteinemia is a risk factor for cardiovascular disease, neurological disorders, and bone abnormalities. The key enzyme in homocysteine metabolism, cystathionine-β-synthase (CBS) is recognized as a target for new homocysteine-lowering therapies including enzyme replacement and gene therapy. Currently, there are no pharmacotherapies available that enhance CBS activity through its allosteric mechanism. The only known allosteric activator of CBS is S-adenosyl-L-methionine (SAM), which is available as a food supplement, but its effectiveness is limited by low membrane permeability and universal involvement in methylation reactions as a substrate. The discovery of CBS activators in high-throughput screening is challenging due to a lack of dedicated assays. Available HTS-compatible activity assays for CBS rely on measuring the product hydrogen sulfide or methanethiol where the signal increases with increased CBS activity. In the case of fluorescence-based assays, it is challenging to discern activators from autofluorescent compounds. In this study, we introduce a homocysteine consumption assay for isolated human CBS (HconCBS) based on the absorbance of Ellman's reagent. This assay leverages a decrease in signal upon CBS activation, with performance parameters exceeding the requirements for high-throughput screening. In a commercial library of 3010 compounds, the HconCBS assay identified 10 hit compounds as more active than SAM, whereas a fluorescence-based assay using 7-azido-4-methylcoumarin (AzMC) identified 141 hits. HconCBS identified 101 compounds with autoabsorbance which did not include hit compounds, while the fluorescence-based assay identified 383 autofluorescent compounds which included all hit compounds. While 4 out of 10 HconCBS hits were confirmed when purchased from a new source, the compounds affected homocysteine rather than CBS. Nevertheless, HconCBS consistently detected the CBS activator seleno-adenosyl-L-methionine (SeAM) added to 4 library plates and re-discovered the same library hits in 3 out of 4 re-screened plates. Taken together, HconCBS was designed to enable the discovery of allosteric CBS activators with greater reliability than fluorescence-based methods. Despite identifying some compounds that acted on homocysteine rather than CBS, the assay consistently identified the CBS activators SAM and SeAM and demonstrated reproducibility across two screening rounds. These findings establish HconCBS as a valuable tool for identifying potential therapeutic candidates for hyperhomocysteinemia, addressing a key gap in the development of CBS-targeted pharmacotherapies.
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Problems of energy security, diversification of energy sources, and improvement of technologies (including alternatives) for obtaining motor fuels have become a priority of science and practice today. Many scientists devote their scientific research to the problems of obtaining effective brands of alternative (reformulated) motor fuels. Our scientific school also deals with the problems of the rational use of traditional and alternative motor fuels.This article focused on advances in motor fuel synthesis using natural, associated, or biogas. Different raw materials are used for GTL technology: biomass, natural and associated petroleum gases. Modern approaches to feed gas purification, development of Gas-to-Liquid-technology based on Fischer–Tropsch synthesis, and liquid hydrocarbon mixture reforming are considered.Biological gas is produced in the process of decomposition of waste (manure, straw, grain, sawdust waste), sludge, and organic household waste by cellulosic anaerobic organisms with the participation of methane fermentation bacteria. When 1 tonne of organic matter decomposes, 250 to 500–600 cubic meters of biogas is produced. Experts of the Bioenergy Association of Ukraine estimate the volume of its production at 7.8 billion cubic meters per year. This is 25% of the total consumption of natural gas in Ukraine. This is a significant raw material potential for obtaining liquid hydrocarbons for components of motor fuels.We believe that the potential for gas-to-liquid synthetic motor fuels is associated with shale and coalfield gases (e.g. mine methane), methane hydrate, and biogas from biomass and household waste gases.