Applied Biological Chemistry最新文献

Kale (Brassica oleracea) is a cruciferous vegetable known for health benefits, primarily due to sulforaphane, a compound with notable anticancer properties. Recently, we reported that the application of geraniol can significantly increase sulforaphane content in kale. However, the molecular mechanisms underlying this enhancement remain unexplored. In this study, we aimed to elucidate the metabolic pathways by which geraniol influences sulforaphane biosynthesis. Geraniol was applied as a 500-ppm solution, sprayed on the leaves of kale cultivated in an indoor aeroponic farm, once daily for two consecutive days, one week prior to harvest. As a result, we found that the sulforaphane content in the geraniol-treated group was 2.2 times higher than in the control group. Moreover, gene expression analysis demonstrated significant upregulation of CYP83, SUR1, and UGT, which are key genes involved in sulforaphane biosynthesis, with increases of 1.7, 1.3, and 1.1-fold, respectively, in the geraniol-treated group compared to the control group. These findings suggest that geraniol enhances sulforaphane content by upregulating genes critical to its biosynthesis.

Dysbiosis of the gut microbiota has increasingly been associated with atopic dermatitis (AD), a chronic inflammatory dermatological disorder. Butyrate, a short-chain fatty acid recognized for its significant anti-inflammatory capabilities, has garnered particular interest among gut microbial metabolites. The process that converts acetoacetyl-CoA to 3-hydroxybutyryl-CoA, which is important for making butyrate, is carried out by L26HBD, an enzyme from F. prausnitzii L2-6 that depends on NAD⁺. We determined the crystal structure of L26HBD in association with NAD⁺ and acetoacetyl-CoA to clarify the structural information of its catalytic action. The monomeric enzyme consists of two distinct domains: a C-terminal domain responsible for dimerization and an N-terminal Rossmann fold that binds NAD⁺. The enclosure of the active site arises from a significant conformational shift in the clamp-lid domain induced by substrate binding, with a root-mean-square deviation of 2.88 Å. The induced fit mechanism was corroborated by structural comparisons between the ligand-free and substrate-bound forms, revealing substrate-driven cavity contraction. Despite the identification of the acetoacetyl-CoA binding mechanism, electron density and B-factor measurements indicated that it exhibited lower stability compared to NAD⁺ binding. These findings enhance our understanding of butyrate biosynthesis in commensal gut bacteria by providing mechanistic insights into substrate detection and catalysis by L26HBD.

We investigated microbial community dynamics and potential pathogen persistence across treatment stages in a livestock manure treatment plant (LMTP) and a municipal wastewater treatment plant (WWTP). To quantify taxon-specific absolute abundances, we used quantitative microbiome profiling (QMP) by combining 16 S rRNA gene amplicon sequencing with quantitative PCR (qPCR) and 16 S rRNA gene copy number (16 S GCN) correction. The LMTP influent was enriched with fermentative and methanogenic taxa such as Candidatus Cloacamonas and Methanobrevibacter and showed a consistent decline in the total microbial load through successive stages. In contrast, the WWTP influent was dominated by gut-associated bacteria, including Lactococcus and Segatella. During sludge treatment and anaerobic digestion, genera such as Xylanibacter and Methanothermobacter became enriched. Functional predictions indicated that anaerobic chemoheterotrophy generally predominated from influent to effluent in both facilities. Although total microbial abundance decreased substantially in the final effluents, potentially pathogenic genera such as Moraxella, Acinetobacter, and Mycobacterium remained detectable at low levels. Integrating absolute abundance data with microbial profiling revealed significant taxon shifts that were missed by relative abundance analysis. These findings provide a more robust basis for assessing microbial dynamics and pathogen persistence, and offer practical insights for risk-based management of wastewater treatment processes.

Although space exploration has immense untapped potential, the harsh conditions of outer space pose severe threats to the physiological and mental health of astronauts. Microgravity and space radiation can have a multitude of biological effects, ranging from adverse physiological changes to alterations in gene expression. Therefore, increasing amounts of biological research are urgently needed to devise countermeasures for the astronauts during long-term space missions. In space biology research, Caenorhabditis elegans (C. elegans) offers several advantages over other animal models because of its small size, short lifespan, genetic traceability, and lack of ethical complications. In this review, we summarize the equipment and space research conducted thus far using C. elegans. Several biological alterations caused by environmental conditions in space have been identified, such as genomic, epigenomic, metabolic, muscular and neuromuscular, immunological, neuronal, and longevity changes, thus highlighting the applicability of C. elegans as a model organism. In addition, we explore the feasibility of integrating chemical analysis into space research, as well as incorporating biopharmaceuticals and nutraceuticals in the treatment of spaceflight-associated disorders.

Constitutive promoters such as CaMV 35S and ubiquitin are commonly utilized in crop genome editing. However, their ectopic overexpression patterns may lead to off-target effects. To address this limitation, tissue-specific or developmentally regulated promoters offer promising alternatives. The RIBOSOMAL PROTEIN S5A (RPS5A) promoter has demonstrated superior editing efficiency compared to the 35S and ubiquitin promoters in dicotyledonous species, yet its potential application in monocots remains unexplored. In this study, we identified and functionally characterized the Oryza sativa RPS5 (OsRPS5) promoters and evaluated their utility in CRISPR/Cas9-mediated genome editing. The activities of the OsRPS5 promoters were assessed through GFP reporter expression in rice protoplasts, and their genome editing capability was validated by targeting two endogenous genes, OsPDS and OsBADH2. Genome editing driven by the OsRPS5 promoter targeting OsPDS resulted in albino phenotypes in approximately 50% of the transgenic lines, with insertion/deletion mutations confirmed through sequencing analysis. Notably, the genome editing efficiency driven by the OsRPS5 promoter was comparable to that of the widely used constitutive promoters in monocots. These findings suggest the OsRPS5 promoter as a potentially more precise and efficient alternative to constitutive promoters for genome editing applications in monocot crops.

Background

Amidst the rising global prevalence of diabetes, exploring novel anti-diabetic agents remains a crucial endeavor. This study investigated the biochemical mechanism of action of a poly-herbal formulation (PHF5) on HepG2 cell lines as well as molecular interactions between bioactive compounds of PHF5 and PKB/Akt, AMPK. PHF5 was formulated from leaves of Ocimum gratissimum, Vernonia amygdalina, Gongronema latifolium, Gnetum africanum, and Aloe barbadensis.

Method

The study employed an experimental design encompassing both in vitro and in silico analysis. HepG2 cells were treated with PHF5 in in vitro studies that looked at parameters like cell viability, glucose uptake, and lipid accumulation. Also, glycation and fructosamine formation were studied in bovine serum albumin (BSA) that had been exposed to fructose and PHF5. In silico investigations utilized virtual screening and molecular docking simulations to elucidate the interactions of phytochemicals from PHF5 with key target enzymes involved in glucose metabolism.

Results

It was found that PHF5 contained key phenolics such as quercetic, rutin etc. through HPLC profiling. In silico modeling demonstrated favorable binding of rutin and quercetin in PHF5 to PKB/Akt and AMPK, key proteins involved in glucose metabolism. The finding here suggests an antidiabetic action of PHF5, which is mediated via activation of the P13K/Akt pathway leading to trafficking of GLUT4 and simulation of insulin secretion. The findings also revealed significant enhancements in cell viability and glucose uptake, coupled with reduced lipid accumulation in HepG2 cells following treatment with PHF5. Additionally, PHF5 demonstrated a mitigating effect on glycation and fructosamine formation.

Conclusion

This study sheds light on the diverse phytochemical composition of PHF5, highlighting potential interactions with crucial enzymes involved in glucose metabolism. The observed promising outcomes points at the potential of PHF5 as a valuable anti-diabetic agent.

The widespread use and improper disposal of plastics in the environment lead to microplastic (MP) pollution. Polyethylene terephthalate (PET) plastics are widely used as single-use plastics, and the mass use of these plastics is contaminating aquatic and terrestrial environments. The transportation of those plastic fragments on agricultural land increases the risk to crop production and food safety. Therefore, the study aimed to evaluate the effect of polyethylene terephthalate microplastics (PET-MPs) on plant growth, nutrient uptake, and physiological stress responses. A short-term effect of PET-MPs (0.1 g/L) on plant growth was assessed using radish (Raphanus sativus) and carrot (Daucus carota var. sativa) grown in half-strength Hoagland solution for one week. PET-MPs did not significantly affect plant biomass and nutrient uptake by plants. Micronutrients such as Cu, Fe, Mn, and Zn were mostly increased in roots and decreased in shoot samples of both plants with PET-MP treatment compared to the control. Although short-term exposure of plants to PET-MPs did not significantly affect plant biomass and nutrient uptake, a significant difference was observed in the physiological stress responses. Chlorophyll a and b contents were significantly (p < 0.05) decreased in radish leaves after PET-MP treatment. Malondialdehyde (MDA) content in the leaves of radish plants significantly increased, indicating that the plant was facing abiotic stress in PET-MP treatment. This study advances understanding of MP-induced phytotoxicity and highlights its potential implications for food safety in agroecosystems.

Strychnine, an alkaloid recognized for its antagonist effects on glycine and acetylcholine receptors, is highly toxic and banned in several countries. It is also set to restricted in Republic of Korea from 2024, creating a need for a quantitative test method to detect strychnine in livestock and fishery products. Therefore, this study aims to develop an analytical method to support the safety management of strychnine. The proposed method extracts and purifies strychnine from livestock and fishery products using ethyl acetate with 2% ammonium hydroxide and primary–secondary amine, respectively. The method demonstrated high sensitivity, with a coefficient of determination exceeding 0.99 and a limit of quantification of 0.005 mg/kg. Average recoveries ranged from 84.7 to 112.9%, with a coefficient of variation below 13.1%. These results meet the Codex guidelines (CAC/GL 71–2009). The developed method was employed to monitor strychnine residue levels in livestock and fishery products, but no residues were detected. The developed method can aid in the quality management of domestically distributed livestock and fishery products.

In this study, we investigated the bioavailability of MK-7 from Bacillus subtilis natto extract Vitamin K2 using the HyperCelle method in a randomized, double-blind, controlled, parallel-group clinical trial. HyperCelle Vitamin K2 showed an average diameter of 321.4 nm with a polydispersity index (PI) of 0.210 and exhibited a population of spherical nanoparticles with a well-defined core-shell structure using dynamic light scattering (DLS) and transmission electron microscopy (TEM), respectively. Its water solubility was higher than that of the powdered form of Bacillus subtilis natto extract Vitamin K2 (negative control). The plasma concentration of Menaquinone-7 (MK-7) from HyperCelle Vitamin K2 was significantly higher than that of the negative control group at 6-, 10-, 24-, and 72-hours post-ingestion (p < 0.05). Both AUC (106.407 ± 73.794 µg·h/mL) and Cmax (3.038 ± 2.184 µg/mL) of MK-7 from HyperCelle Vitamin K2 were significantly greater compared to the negative control (AUC: 36.216 ± 18.985 µg·h/mL, Cmax: 1.474 ± 0.696 µg/mL) (p < 0.05). These results indicate that HyperCelle Vitamin K2, which was well encapsulated with a smaller particle size, enhanced water solubility and consequently improved the bioavailability of MK-7. Given the well-recognized role of MK-7 in supporting bone health, the improved absorption observed in this study suggests potential benefits for biological activity, although further validation is needed before confirming its efficacy in functional food applications.

Consumption of wild plants is a common practice globally; however, not all plants are safe for human consumption, as some are toxic. Toxic plants often resemble edible species, which makes their identification difficult, particularly for non-experts. Convallaria majalis, a poisonous plant containing cardiac glycosides, poses a significant danger if mistakenly ingested. Allium microdictyon is a popular edible wild vegetable in East Asia. Owing to their similar appearance, accidental mixing can occur during harvesting or processing. In this study, specific primer pairs were designed to target chloroplast genes to distinguish the edible plant Allium microdictyon from the toxic plant Convallaria majalis, and their practicality was tested. The specificity, sensitivity, and applicability of the quantitative real-time PCR assay were evaluated using all primer pairs. Six primer sets (three for A. microdictyon and three for C. majalis) exhibited strong linearity, with correlation coefficients exceeding 0.98 and slopes ranging from − 3.22 to − 3.56. PCR efficiencies ranged from 90.80% to 97.85%. Cycle threshold (Ct) values corresponding to 0.1% of the binary mixture were used as the cut-off values. Additionally, the specificity of the primer pairs was validated by analyzing 13 non-target plant species and assessed for practicality using 15 commercial samples. The developed primer pairs will aid in preventing the misidentification of toxic wild plants and serve to guide the identification of toxic species, thereby contributing to public health and safety.