Applied Biochemistry and Biotechnology最新文献

Preeclampsia (PE) is a pregnancy-related syndrome that can lead to a variety of pathophysiological processes, such as impaired implantation. The pathogenesis of PE involves circular RNA (circRNA). The study aims to determine the role of a novel circRNA, circ_0003314, in trophoblast cell phenotypes. Circ_0003314, microRNA-26b-5p (miR-26b-5p) and IL-1 receptor accessory protein (IL1RAP) expression were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation was investigated by MTT assay and 5-Ethynyl-2'-deoxyuridine assay. Cell migration and invasion were investigated by transwell assay. Cell apoptotic rate and angiogenesis were investigated by flow cytometry analysis and tube formation assay, respectively. Protein expression was detected by western blotting. The binding relationship between miR-26b-5p and circ_0003314 or IL1RAP was identified using dual-luciferase reporter assay and RNA pull-down assay. Circ_0003314 and IL1RAP expression were significantly increased, while miR-26b-5p was decreased in placental tissues of PE patients. Circ_0003314 overexpression inhibited trophoblast cell proliferation, migration, invasion and angiogenesis and induced cell apoptosis. Additionally, circ_0003314 acted as a sponge for miR-26b-5p, and miR-26b-5p bound to IL1RAP. Introduction of miR-26b-5p or silencing of IL1RAP attenuated the effects of circ_0003314 overexpression on trophoblast cell phenotypes. Further, circ_0003314 induced IL1RAP expression through miR-26b-5p in trophoblast cells. Circ_0003314 regulated trophoblast cell phenotypes by increasing IL1RAP expression through binding to miR-26b-5p.

The green synthesis of copper oxide nanoparticles (CuO) mediated by crude ethanolic extract and the n-butanol fraction of Adiantum venustum represents a groundbreaking approach in nanotechnology, combining ecological sustainability with advanced functionality. This innovative method leverages the natural bioactive compounds present in A. venustum to produce CuO nanoparticles, which exhibit remarkable antioxidant, anti-inflammatory, antimicrobial, and anti-proliferative properties. The green synthesized nanoparticles were characterized using a variety of techniques, as XRD confirmed the crystalline nature of the CuO nanoparticles, with a crystallite size of 14.65 nm for CuO-C and 18.73 nm for CuO-B. The grain sizes of CuO-C (14.09 ± 0.17 nm) and CuO-B (67.88 ± 2.08 nm) were determined using transmission electron microscopy micrographs. Furthermore, the synthesized nanomaterial and the crude ethanolic extract, n-butanol fraction, were examined for their biological potentials namely antioxidant, anti-inflammatory, antimicrobial, and anti-proliferative activity against HeLa cancer cells. Among the synthesized nanomaterials, copper oxide nanoparticles synthesized by utilizing the n-butanol fraction have appeared as a potential biomedical agent. CuO-B has arisen as an antioxidant agent with IC₅₀ values of 44.63 ± 0.49 µg/mL, 48.49 ± 0.17 µg/mL, and 35.39 ± 0.61 µg/mL for DPPH, FRAP, and reducing power assay, respectively. Furthermore, the significant antibacterial potential of CuO-B against gram-positive (S. aureus MIC 46.88 µg/mL) and gram-negative (K. pneumonia MIC 23.48 µg/mL) bacterial strains cannot be neglected either. Along with this, the IC₅₀ value (138.07 µg/mL) of CuO-B against HeLa cells proved it to be a potential anticancerous agent. Hence, this novel approach emphasized that these synthesized nanoparticles have tremendous biological potential and can be applied to various fields of agriculture and biomedicine.

D-tagatose is a valuable rare sugar with potential health benefits such as antiobesity, low-calorie, prebiotic, and anticancer. However, its production is mainly depending on chemical or enzymatic catalysis. Herein, a cobalt-based metal-organic framework (MOF) was developed at room temperature in an aqueous system using a self-assembly method. The L-arabinose isomerase (L-AI) was immobilized into this unique MOF by an in situ encapsulation process. The morphology and structural aspects of the MOF preparations were characterized by different analytical techniques such as scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), confocal laser scanning microscopy (CLSM), Fourier transform infrared spectroscopy (FT-IR), and X-Ray diffraction (XRD). Moreover, thermogravimetric analysis (TGA) suggested the high thermal stability of the L-AI@MOF. Significantly, the immobilized catalyst exhibited enhanced catalytic efficiency (k_cat/K_m) of 3.22 mM^-1 s^-1 and improved turnover number (k_cat) of 57.32 s^-1. The L-AI@MOF efficiently catalyzes the synthesis of D-tagatose from D-galactose up to the equilibrium level (~ 50%) of isomerization in heterogeneous catalysis. Interestingly, L-AI@MOF was found stable and reusable for more than five cycles without the requirement of additional metal ions during catalysis. Thus, L-AI stabilized in the MOF system demonstrated a higher catalytic activity and potential guidance for the sustainable synthesis of rare sugar D-tagatose.

Sustainable agriculture and the provision of food for all become dependent on the availability of efficient diagnostic techniques for the prompt identification of plant diseases. Current scientific findings suggest that nanotechnology can positively affect the agrifood industry by reducing the adverse effects of agricultural practices on human health and the environment, increasing food security and productivity, and fostering social and economic justice. Nanomaterials' unique physical and chemical characteristics have made it possible to employ them as cutting-edge, effective diagnostic instruments for various plant infections and other significant disease biomarkers. By creating diagnostic instruments and methods, nanobiosensors significantly contribute to the revolution of farming. In real time, nanobiosensors can detect infections, metabolites, pesticides, nutrient levels, soil moisture, and temperature. This helps with precision farming techniques and maximises resource use. To better address agricultural concerns, we have included the most recent research on the concept, types, applications, commercial aspects, and future scope of nanobiosensors in this review.

A complex of ovotransferrin and lysozyme was directly isolated from egg white using an anti-transferrin antibody-immobilized membrane after antiserum proteins were separated by non-denaturing two-dimensional electrophoresis and transferred onto a membrane. The complex retained lysozyme activity that catalyzes the breakdown of peptidoglycans in the bacterial cell wall at the β1-4 bond between N-acetylmuramic acid and N-acetylglucosamine residues. The activity of the purified lysozyme was suppressed to 6.4% in the presence of 1 μmol Fe²⁺, whereas that of the mixture of the purified lysozyme and ovotransferrin was maintained at 58%. The activity of the purified lysozyme was suppressed to 35% in the presence of 10 nmol Fe³⁺, whereas that of the mixture of the purified lysozyme and ovotransferrin was maintained at 66%. Furthermore, the bacteriolytic activity against Bacillus subtilis of egg white with reduced glycoproteins such as ovotransferrin was assessed, and the bacteriolytic activity was found to be suppressed in the presence of Fe²⁺ and Fe³⁺. This suppression was ions, thereby alleviating the inhibition of lysozyme activity by iron ions. A complex of ovotransferrin and lysozyme is efficient because ovotransferrin effectively captures iron ions near lysozyme. Thus, protein complexes containing enzymes can be applied to control their activity.

Rare ginsenosides Rg3 and Rh2, which exhibit diverse pharmacological effects, are derivatives of protopanaxadiol (PPD). UDP-glycosyltransferases, such as the M315F variant of Bs-YjiC (Bs-YjiCm) from Bacillus subtilis and UGTPg29 from Panax ginseng, can efficiently convert PPD into Rh2 and Rh2 into Rg3, respectively. In the present study, the N178I mutation of Bs-YjiCm was introduced, resulting in an increase in Rh2 production. UDP-glycosyltransferase UGTPg29 was then engineered to improve its robustness through semi-rational design. The variant R91M/D184M/A287V/A342L, which indicated desirable stability and activity, was utilized in coupling with the N178I variant of Bs-YjiCm and sucrose synthase AtSuSy from Arabidopsis thaliana to set up a "one-pot" three-enzyme reaction for the biosynthesis of Rg3. The influential factors, including the ratio and concentration of UDP-glycosyltransferases, pH, and the concentrations of UDP, sucrose, and DMSO, were optimized. On this basis, a fed-batch strategy was adopted to achieve a Rg3 yield as high as 12.38 mM (9.72 g/L) with a final yield of 68.78% within 24 h. This work may provide promising UDP-glycosyltransferase candidates for ginsenoside biosynthesis.

Hepatocellular carcinoma (HCC) is the most common type of liver cancer, characterized by a high morbidity rate. Long non-coding RNAs (lncRNAs) play an important role in regulating various cellular processes and diseases, including cancer. However, their specific roles and mechanisms in HCC are not fully understood. This study used a multi-cohort design to investigate necroptosis-related lncRNAs (NRLs) in patients with HCC. We curated a list of 1095 NRLs and 838 genes showing differential expression between tumor and normal tissues. Among them, we found 105 NRLs closely associated with the prognosis of HCC patients. The 10 lncRNAs (AC100803.3, AC027237.2, AL158166.1, LINC02870, AC026412.3, LINC02159, AC027097.1, AC139887.4, AC007405.1, AL023583.1) generated by LASSO-Cox regression analysis were used to create a prognostic risk model for HCC and group patients into groups based on risk. The KEGG analysis revealed distinct pathway enrichments in high-risk (H-R) and low-risk (L-R) subgroups. According to GO analysis, this study identified 230 differentially expressed genes (DEGs) that were significantly enriched in specific biological processes. Comparison of immune checkpoint-related genes (MCPGs) between H-R and L-R patients revealed significant differences. Moreover, we established a correlation between the risk scores of patients with liver cancer and their sensitivity to 16 chemotherapeutic agents. Employing protein-protein interaction (PPI) analysis, we identified 10 hub genes that potentially regulate the molecular networks involved in HCC development. This study is a pioneering effort to investigate the roles of NRLs in HCC. It opens a new avenue for potential targeted therapies and provides insights into the molecular mechanisms of HCC.

Natural alternatives to synthetic fungicides are gaining prominence as the focus sharpens on developing protective products. In this regard, the present study aimed to discern the antifungal potential of Artemisia absinthium L., a traditional medicinal plant native to Kashmir, by identifying its key phytoconstituents. Here, we present the notable antifungal activity of the leaf extract of A. absinthium against Fusarium oxysporum, Penicillium digitatum, and Alternaria solani, as observed through in vitro tests. Further refinement of the extract through chromatography isolated fraction 4 (F4), unveiling 27 compounds with no prior literature on their occurrence in A. absinthium. Additionally, in silico molecular docking analysis revealed three compounds which include Bruceine B (1), Guanidine, (phenylmethyl)- (2) and Ethyl alpha-d-glucoside (3) as potent inhibitors of multiple target key fungal enzymes such as endopolygalacturonase, chitin deacetylase and 1, 3, 8-trihydroxynaphthalene. The virtual screening unveiled compounds 1-3 within fraction 4, displaying robust binding energy ranging from -8 to -5.8 kcal/mol with multiple target enzymes. Notably, their efficacy surpassed that of the reference commercial fungicide, benodanil. This study underscores the burgeoning interest in harnessing natural alternatives for fungicidal applications, highlighting the potential of A. absinthium as a valuable resource in the quest for sustainable and effective bio-fungicides.

Traditional medicinal plants have attracted scientific interest due to their bioactive compounds, and the levels of their constituents vary with location and altitude. The present study was designed to evaluate the pharmacological potential of two selected traditional medicinal plants, Mikania micrantha and Ageratum houstonianum collected from two sites, Murlen National Park (MNP) and Dampa Tiger Reserve (DTR), located at different altitudes. Both plant species are used by local traditional healers in Mizoram, Northeast India, to treat various health problems. We hypothesized that altitudinal variation would affect these plants' chemical composition and bioactive potential. Plant extracts were evaluated for antioxidant and cytotoxic activities. The results show that the plants located at a higher altitude, i.e., MNP, showed higher TPC (615.7 ± 0.58 and 453.80 ± 0.95 µg gallic acid equivalents/mg of plant extract dry weight (µg GAE/mg) for M. micrantha and A. houstonianum , respectively) and TFC (135.4 ± 0.46 and 120.66 ± 1.93 µg quercetin equivalents/mg of plant extract dry weight (µg GE/mg) for M. micrantha and A. houstonianum, respectively). The extract of A. houstonianum. (MNP) exhibited significantly greater antioxidant activity against ABTS radicals (IC₅₀ 241.6 µg/mL) as compared to the extract of A. houstonianum (DTR) (IC₅₀ 371.2 µg/mL). The composition of the bioactive compounds present in the plants was determined using UPLC-ESI MS/MS and GC/MS, which detected five and ten compounds in the A. houstonianum and M. micrantha extracts, respectively. Plant species collected from the Murlen National Park site had high bioactivity potential and contained several bioactive compounds. A distinct variation between the volatile and non-volatile compounds was revealed. The collective data in this study show the influence of altitude on the biological compound production of selected medicinal plants. The findings will be utilized in the plant material needed for developing bioactive formulations.