Applied Biological Chemistry最新文献

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.

Background

Pneumonia is a serious respiratory disease that significantly impacts human health. Quercetin (QUE), a plant extract, has been shown to exert protective effects against lung injury, but its precise mechanisms remain unclear.

Methods

Lipopolysaccharide (LPS) was used to induce injury in embryonic lung fibroblasts. Cell viability, proliferation, and apoptosis were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, 5-ethynyl-2’-deoxyuridine (EdU) staining, and flow cytometry assay, respectively. Reactive oxygen species (ROS), malondialdehyde (MDA), and Fe²⁺ levels were evaluated using corresponding assay kits. The levels of inflammatory factors were determined by enzyme-linked immunosorbent assay (ELISA). Quantitative real-time PCR (qRT-PCR) and western blot were employed to analyze the expression and correlation of related genes. Methylated RNA immunoprecipitation (MeRIP) was employed to verify the regulatory mechanism between genes.

Results

QUE inhibited embryonic lung fibroblast injury. Both transducin β-like 1 X-linked receptor 1 (TBL1XR1) and methyltransferase 3 (METTL3) were highly expressed in lung injury tissues and cells, which were decreased by QUE treatment. TBL1XR1 deficiency weakened LPS-induced embryonic lung fibroblast injury. Mechanistically, METTL3 mediated the N⁶-methyladenosine (m6A) methylation and stabilized the TBL1XR1 mRNA. TBL1XR1 up-regulation alleviated the retardation of METTL3 absence on embryonic lung fibroblast injury. Furthermore, METTL3 overexpression reversed the inhibition of QUE on embryonic lung fibroblast injury.

Conclusion

QUE inhibited embryonic lung fibroblast injury by regulating the METTL3-mediated m6A methylation of TBL1XR1 mRNA.

The soil-applied pesticide can transfer to cultivated crops, and the residue concentration in the crops is decreased time-dependently by biological decomposition and crop growth. Pencycuron was most popularly used as a fungicide in field soil; however, the residue dissipation in greenhouse soil and its bioconcentration factor (BCF) for spinach, one of the most popular leafy vegetables, were not investigated. To estimate the residue dissipation of the soil-applied pencycuron, the residue dissipation was monitored in the recommended dose and the double dose treatments, and the 50% dissipation times ranged from 50 to 54 days; these times were met by the spinach cultivation time from seedling to harvest. The pencycuron residues in the spinach at the recommended dose and the double dose treatments were 0.006 mg kg⁻¹ and 0.025 mg kg⁻¹ at 48 days after the treatment (DAT), and 0.003 mg kg⁻¹ and 0.008 mg kg⁻¹ at 55 DAT, respectively. From the experimental data, the estimated BCFs of pencycuron were 0.003–0.008 for the spinach at 48 DAT and 0.001–0.002 at 55 DAT. During the seven-day additional cultivation, the fresh weights of the spinach were increased 1.5–1.8 times; thus, the crop growth would be considered as a main factor for the estimation of the crop residue.

This study aimed to analyze the chemical composition of Syrian black seed honey, and then utilized its extract to synthesize silver nanoparticles (AgNPs), and assess their antimicrobial properties. The sugar content of the black seed honey showed fructose, glucose, and sucrose levels at 38.7 ± 1.50 g/100 g, 7.4 ± 2.13 g/100 g and 0.7 ± 0.20 g/100 g, respectively. Total phenolic content (TPC) and total flavonoid content (TFC) were measured at 263.3 ± 0.72 mg GAE /100 g, and 76.41 ± 1.20 mg QE /100 g, respectively. High-performance liquid chromatography (HPLC) confirmed the presence of six compounds: kaempferol, caffeic acid, cinnamic acid, apigenin, quercetin and chrysin. A concentrated solution of the honey, containing sugars, phenolics and flavonoids, was used to synthesize AgNPs, which were characterized using UV–Vis Spectroscopy, Dynamic Light Scattering (DLS) and Field Emission Scanning Electron Microscopy (FESEM). The AgNPs were well-dispersed and stable, with sizes ranging from 3 to 15 nm, with a polydispersity index (PDI) of 0.314 ± 0.02, and a ζ-potential of -21.7 mV according to DLS measurements. Microscopy and UV-Vis spectroscopy confirmed the formation of nanoparticles. The stable AgNPs were spherical with an average size of 26 nm according to FESEM, and the localized surface plasmon resonance (LSPR) peak in the UV-Vis Spectroscopy was observed at 415 nm. The antimicrobial activity of AgNPs was evaluated against Staphylococcus aureus, Staphylococcus epidermidis, Klebsiella spp., Escherichia coli, Pseudomonas aeruginosa, and Candida spp. The inhibition zones were (22 ± 0.28, 25 ± 0.41, 20 ± 0.33, 20 ± 0.27, 23 ± 0.43, and 20 ± 0.20 mm) respectively.