Applied Biochemistry and Biotechnology最新文献

Antiviral lignin, designated as FR₂₀₀, was produced using acidic microwave glycerolysis of sugarcane bagasse. The lignin strongly inhibited infection by the human norovirus surrogate, feline calicivirus (FCV) without substantial cytotoxicity. The antiviral activity emerged through direct contact of the lignin with the virion. To develop antiviral fibers, cotton was coated with the antiviral lignin AIF₂₀₀, a crude fraction obtained in the process of FR₂₀₀ preparation. Autofluorescence microscope images confirmed successful even coating of the lignin on the cellulose fiber surfaces. ATR FT-IR spectra supported the incorporation of the lignin. The strong anti-FCV activity of the cellulose-lignin assembly was observed by measuring viral titers of the filtrate passing through the lignin-immobilized cotton. The antiviral material developed in this study, totally composed of lignocellulosic biomass, is potentially useful for protecting our health by capturing virions in daily life and livestock environments.

The healing process of wounds, as soft tissue injuries, is challenging. Bone marrow mesenchymal stem cells (BMSCs) and exosomes (Ex) they secrete are critical for skin wound healing. Paracrine signaling mechanisms appear to facilitate the therapeutic properties of BMSCs. However, BMSC therapy has not yet been extensively explored in terms of specific cellular interactions between macrophages and BMSCs. In this study, macrophage depletion had a substantial negative impact on the wound healing capabilities of BMSCs, highlighting macrophages' crucial role in facilitating wound healing processes mediated by BMSCs. BMSCs transferred to the wound site promoted M2 polarization and alleviated wound healing. The co-cultivation of BMSCs and macrophages resulted in an enhanced phenotypic polarization towards M2. A mechanistic explanation for this effect was found in BMSCs-Ex. Additionally, miR-153-3p, derived from BMSCs-Ex, was identified as a regulator of macrophage polarization via its targeting of KPNA5. Through the transfer of BMSCs-Ex-derived miR-153-3p, BMSCs are capable of promoting M2 polarization and can potentially facilitate wound healing.

Mesenchymal stem cells (MSCs) and their secreted exosomes (Exos) have attracted much interest for their potential against ischemia/reperfusion injury (IRI). In the present research, we employed rat ovarian ischemia/reperfusion injury (OIRI) model and hypoxia/reoxygenation (H/R) model of primary rat ovarian granulosa cells (RGCs) to investigate whether BMSC-Exos could alleviate OIRI. Our data suggested that administration of BMSCs in rats significantly reduced OIRI-resultant histopathological changes, oxidative stress, inflammation, and pyroptosis. In addition, BMSCs downregulated the expression of proteins related to the TXNIP/NLRP3 inflammasome pathway. Based on in vitro experiments, BMSC-Exos could be internalized by RGCs and curbed oxidative stress and pyroptosis in H/R-injured RGCs. This effect may be due to the regulation of TXNIP/NLRP3 inflammasome pathway. Remarkably, our in vivo data were in concordance with our in vitro results, suggesting that BMSC-Exos suppressed OIRI-induced oxidative stress and pyroptosis via the TXNIP/NLRP3 inflammasome pathway. Overall, these results demonstrate good therapeutic efficacy of BMSC-Exos for treating OIRI, which may provide experimental evidence for the potential clinical benefits of BMSC-Exos for ovarian protection.

The metabolic enzyme ATP citrate lyase is overexpressed in several cancers and links glucose metabolism with de novo fatty acid synthesis pathway by catalyzing the conversion of citrate into acetyl CoA and oxaloacetate. Potassium hydroxycitrate, its natural inhibitor, exhibits anticancer activity; however, its use is limited due to low bioavailability. This study aims to improve the efficacy of hydroxycitrate by its encapsulation in bovine milk exosome surface conjugated with folate for targeting lung cancer cells. The mean particle size of potassium hydroxycitrate-loaded exosomes (Exo-KH) and paclitaxel exosomes (Exo-Pac) was 183 nm and 174 nm; they had spherical morphology and encapsulation efficiency of 16.87 ± 2.78% and 27.65 ± 3.23%, respectively. In the in vitro study, Exo-KH suppressed the proliferation of A549 cells and significantly reduced ACLY mRNA expression. In addition to ACLY, EXO-KH also downregulated the mRNA expression of other crucial metabolic enzymes such as fatty acid synthase and isocitrate dehydrogenase 1. EXO-KH formulation caused significant increase in apoptosis rate (< 75%) and reactive oxygen species production and reduced ACLY protein expression in A549 cells. Moreover, the pharmacokinetic study revealed the sustained release of hydroxycitrate (half-life 22.74 h and clearance 0.13 µg/ml) from the exoformulation. Altogether, the study findings highlight the beneficial role of EXO-KH formulation against lung cancer.

Flavonoids are known to possess biological effects like anti-inflammatory, antibacterial, antioxidant, and antidiabetic properties. Similarly, silver nanoparticles (AgNPs) have been widely used in the biomedical industry for therapy and diagnostics for a long time. This study investigates the potential of naringenin functionalized silver nanoparticles (AgN NPs) as a potential wound healing agent. The synthesis of AgN NPs was carried out using the one-pot synthesis method in the alkaline pH. Naringenin is used as the capping and the reducing agent. The naringenin-capped AgNPs were synthesized in six different concentrations. The structural, morphological, and spectroscopic characterization for each sample was conducted. The size of the nanoparticles was studied using the dynamic light scattering (DLS) experiment and further confirmed using TEM. The crystalline structure was investigated using X-ray diffraction, and AgN NPs exhibited a fcc crystal structure. The FTIR confirmed the capping of naringenin on AgNPs. All samples were tested for antibacterial activity, and the results demonstrated zones of inhibition against both Gram-positive Staphylococcus aureus and Gram-negative bacteria, such as Escherichia coli and Pseudomonas aeruginosa. Also, AgN NPs exhibited dose-dependent anti-inflammatory, antioxidant, and antidiabetic properties. The wound healing potential of AgN NPs was evaluated using a scratch wound assay in L929 cell lines. After 24 h, the scratch area was significantly reduced in the AgN NPs-treated sample, indicating enhanced cell migration compared to naringenin. Hence, these findings suggest that AgN NPs may serve as a more promising wound-healing agent than naringenin.

Colorectal cancer (CRC) is one of the common deadliest cancers worldwide. In Malaysia, the numbers of new CRC cases were horrific and worrisome. Telomerase is both prognostic indicator and predictor of carcinogenesis in CRC patients. Berberine, a telomerase inhibitor, was used in clinical trials and metabolomic studies; however, the association of telomerase with metabolites and metabolic pathways was not fully understood. Colorectal cancer cell line HCT 116 was cultured and treated with 10.54 µg/mL berberine. The cells were harvested at different time points to conduct subsequent analyses. The methods used in this research were real time-polymerase chain reaction (RT-PCR) to assess RNA expressions; Western blot to determine protein levels; TELOTAGGG Telomerase PCR ELISA to determine relative telomerase activity (RTA); 4',6-diamidino-2-phenylindole (DAPI) staining to determine percentage of nuclei damage; fluorescence microscopy for cell area; spectrophotometric potassium iodide assay for intracellular hydrogen peroxide concentration [H₂O₂]; as well as liquid chromatography mass spectrometry (LCMS) and tandem mass spectrometry (MS/MS) to investigate the intracellular metabolites. Partial least square-discriminant analysis (PLS-DA) score plot exhibited an improved separation compared to principal component analysis (PCA) when metabolomic data analysis of HCT 116 at various berberine treatment durations was conducted. Time and berberine treatment had an impact on RTA in HCT 116. RTA was discovered to be positively and negatively correlated to 14 and 2 metabolites, respectively. Glutamic acid was consistently found correlated to RTA. Other four metabolites, i.e., MG(14:0), [3-[hydroxy(phosphonooxy)phosphoryl]oxyphenyl] phosphono hydrogen phosphate), (3S,6S)-6-[[(3S,6R)-6-[(2S,3S,5S)-2,5-diiodo-4-methoxy-6-methyloxan-3-yl]oxy-3,4,5-trihydroxyoxan-2-yl]methoxy]-3,4,5-trihydroxyoxane-2-carboxylic acid, and 1-[5-O-(5'-adenylyloxyphosphonyl)-beta-D-ribofuranosyl]-5-amino-1H-imidazole-4-carboxamide, were newly discovered to be connected to RTA in HCT 116. Four metabolic pathways that majorly affected shared glutamic acid and glutamine. Nitrogen metabolism, D-glutamine and D-glutamate metabolism, glyoxylate and dicarboxylate metabolism, and aminoacyl-tRNA biosynthesis have been identified to be associated with RTA. Network analyses hinted that glutamic acid was also associated with oxidative stress mechanism. The multiple roles glutamic acid acted in diverse metabolic pathways and interaction networks emphasized the importance of glutamic acid in HCT 116 regarding RTA. This research establishes the association between RTA and several chosen RNAs, proteins, metabolites, and oxidative stress mechanisms, consequential in morphological alteration in HCT 116, to expand the knowledge of the intricate biological relationships and telomerase mechanism in CRC.

Sepsis is a serious systemic infection with a high mortality rate. More and more evidence suggested that necroptosis plays a crucial role in the pathogenesis and progression of sepsis. This study aimed to elucidate the biological function and clinical significance of necroptosis in sepsis, and identify new potential biomarkers to improve the diagnosis and treatment of sepsis. Firstly, we identified 40 differentially expressed necroptosis related genes (DENRGs). Subsequently, a protein interaction (PPI) network of 40 DENRGs was constructed. Based on the key NRGs in the PPI network, the LASSO algorithm was used to screen eight diagnostic-related NRGs in sepsis, and a diagnostic model and risk score were constructed. The ROC analysis results indicated that the eight NRGs diagnostic model has good diagnostic performance (AUC = 0.955). There is a significant difference in risks core between normal samples and sepsis patients. The results of immune infiltration analysis showed that eight diagnostic-related NRGs were significantly correlated with multiple immune cells. Given the clinical significance of necroptosis in sepsis, we identified two molecular subtypes of sepsis based on eight NRGs. The necroptosis score of subtype 1 is significantly lower than that of subtype 2, while the immune score of subtype 1 is significantly higher than that of subtype 2. In summary, we developed and validated a diagnostic model and risk score based on eight NRGs, and identified two completely different subtypes associated with sepsis. Our research may provide new insights into the mechanisms of necroptosis in sepsis and the identification of potential biomarkers.

Fruit wastes contain significant levels of polyphenols with various health-related effects. However, polyphenols have several limitations that hinder their industrial applications. In this study, solid-state fermentation was used as a pre-extraction treatment to not only increase the extraction efficiency of polyphenols from watermelon peel (WP), but also to overcome their drawbacks and improve other properties. Specifically, optimal conditions in the integrated process, fermentation and extraction, were determined by response surface methodology for the highest polyphenol recovery from WP. The post-process extract showed a dramatic improvement in polyphenol content with an increase in various phenolic compounds determined by UHPLC-MS/MS and a significant decrease in cytotoxicity. In addition, two limitations of polyphenols, low stability and bioaccessibility, were significantly improved. Various health potentials of WP were also enhanced: antioxidant (radical scavenging and ferric reduction), anti-inflammatory (inhibition of heat-induced hemolysis, protein denaturation and protease), selectively antibacterial (anti-growth, anti-survival and anti-biofilm for pathogenic bacteria), prebiotic (promotion of probiotic growth and biofilm formation), anti-diabetic (α-glucosidase and α-amylase inhibition), and anti-melanogenic (tyrosinase inhibition). The mechanisms of enzyme inhibition for anti-diabetic and anti-melanogenic capacity were further elucidated by enzyme kinetic studies. Overall, the study suggests an efficient process to convert WP into high-value products with prospects in the circular economy.

This study aimed to investigate how Tanshinone IIA (Tan IIA) affects gastric cancer cell (MGC803) proliferation under anaerobic conditions, which are linked to drug resistance and tumor growth. The proliferation of MGC803 cells under both aerobic and anaerobic conditions in response to Tan IIA was assessed using the Cell Counting Kit-8 (CCK-8) assay. To elucidate the molecular mechanisms underlying these effects, proteomics analysis was performed following treatment with 50 µmol/L Tan IIA, focusing on alterations in Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Additionally, in vitro evaluations such as glucose uptake, lactate production, and adenosine triphosphate (ATP) synthesis were employed to validate the alterations in glycolytic activity observed in anaerobic cells treated with Tan IIA. Under anaerobic conditions, Tan IIA enhanced the inhibitory effect on the proliferation of MGC803 cells. Proteomics data revealed that a total of 6629 proteins were identified and quantified using liquid chromatography-tandem mass spectrometry (LC-MS/MS), with 2604 proteins exhibiting significant changes (fold change > 2 or < 0.5, P < 0.05). KEGG analysis highlighted the perturbation of glycolytic pathway by Tan IIA under anaerobic conditions, accompanied by reduced glucose uptake, lactate production, and ATP synthesis. Additionally, a downregulation of glycolytic enzyme expression was observed at both the mRNA and protein levels, including glyceraldehyde-3-phosphate dehydrogenase (GAPDH), lactate dehydrogenase A (LDHA), phosphofructokinase 2 (PFKP), and pyruvate dehydrogenase (PDH). Tan IIA inhibits the proliferation of MGC803 cells by disrupting the glycolysis under anaerobic conditions, offering a potential treatment for anaerobiosis-resistant solid tumors.