Applied Biochemistry and Biotechnology最新文献

Bean common mosaic virus (BCMV) is a severe plant pathogen of common bean (Phaseolus vulgaris L.), that causes huge yield losses across the globe. The virus has a wide host range and varied modes of transmission, due to which its management is challenging. Pathogen-derived resistance, which entails inserting virus-derived gene sequences into transgenic plants, is extremely effective in overcoming plant viruses. However, owing to ethical and biosecurity concerns, transgenic crops have not been widely accepted. Exogenous application of double-stranded RNA (dsRNA) is a new and intriguing method for inducing resistance against plant viruses. In this study, the efficacy of exogenous application of dsRNAs synthesized from BCMV helper component proteinase (HC-Pro) and coat protein (CP) genes were assessed in three plants: tobacco (Nicotiana tabacum), common bean (Phaseolus vulgaris L.), and cowpea (Vigna unguiculata), and both dsRNAs elicited a resistance response. dsRNA targeting the HC-Pro gene of BCMV was found more effective in inducing RNAi-based resistance than dsRNA targeting the CP gene of BCMV with a more pronounced effect in cowpea than tobacco and common bean. We showed the stability and transport of both dsHC-Pro and dsCP in inoculated to non-inoculated young leaves. We also showed the ability of mesoporous silica nanoparticles (MSP) conjugated with dsHC-Pro to provide prolonged stability and broader resistance against BCMV in common bean, extending protection from 12 dpi up to 20 dpi, compared to naked dsHC-Pro. Our results suggest that dsRNA produced from HC-Pro and CP genes of BCMV can induce RNAi-based resistance against BCMV infection.

Medicinal plants have long been recognized as a valuable source of human health due to their therapeutic potential in treating a variety of diseases. Tarenna alpestris, a plant native to the Megamalai Hills in the Western Ghats of India, has traditionally been used for numerous medicinal purposes. However, despite its extensive use in folklore, scientific validation of its therapeutic properties remains limited. This study aims to evaluate the phytochemical composition of Tarenna alpestris, assess its antioxidant properties, and explore its potential anticancer effects against HT-29 colon cancer cells. The phytochemical profile was determined using preliminary screening and gas chromatography-mass spectrometry (GC-MS). Antioxidant activity was measured through DPPH, FRAP, H₂O₂, and N₂O₂ assays. The anticancer effects were investigated using the MTT assay for cell viability, AO/EtBr staining for apoptosis detection, DAPI staining for nuclear fragmentation analysis, and flow cytometry for cell cycle analysis. The phytochemical analysis identified several bioactive compounds, including flavonoids, alkaloids, terpenoids, and phenolic acids. Sixteen phytocomponents were detected from the extract by GCMS analysis, the major compounds are 9-octadecynoic acid (21%), N-hexadecanoic acid (16%), methylene diamine, N,N'-diacetyl (15%), 1-allyl-cyclohexane-1,2-diol (11%) 2-methyl-6-methylene-octa-1,7-dien-3-ol (5%), squalene (4%), and lupeol (3%) respectively. The antioxidant assays demonstrated significant free radical scavenging activity, with IC50 values comparable to known antioxidant standards. The antioxidant enzymatic activity of Tarenna alpestris extract suggests a potent ability to neutralize reactive oxygen species and protect against oxidative damage. In vitro studies revealed that Tarenna alpestris extract significantly inhibited the proliferation of HT-29 colon cancer cells and induced apoptosis, based on concentration dependent manner. The concentration needed to inhibit 50% of cell growth, known as the IC₅₀ value, was found to be 26 ± 0.20 μg/mL. Additionally, cell cycle analysis showed G0/G1 phase arrest in treated cells. Tarenna alpestris exhibits a robust phytochemical profile with substantial antioxidant and anticancer properties. These findings support its potential as a therapeutic agent for cancer prevention and treatment. Further research, including in vivo studies, is warranted to fully elucidate its therapeutic efficacy and mechanisms of action.

Background: Solanum melongena Linnaeus (brinjal) belongs to the Solanaceae family and is also known as eggplant. It is one of the most common vegetables that is grown abundantly and consumed by a large number of people. However, it is found to be highly susceptible to harmful pests such as brinjal shoot and fruit borer, (Leucinodes orbonalis) which are responsible for causing severe damage to the plant's health and, correspondingly, its yield. Damages include shoot and leaf spoilage which leads to overall hampering of the metabolic process of the plant. This study aims to suggest that the plant, Solanum melongena L., has certain self-induced mechanisms to withstand these stress and pest attacks by secreting compounds known as "Chlorogenic Acid." Chlorogenic acid is known to be a plant-derived product and is a part of secondary metabolites. Different plant parts were examined for their diverse secondary metabolite content under laboratory conditions. The study was further proceeded by implementing chlorogenic acid exogenously, on the pest-infected plants at a concentration of 1 mg/ml in two different foliar sprays, one consisting of simple water and another 50% ethanol. Moreover, molecular analysis shows a higher expression of the genes which are pivotal for the secretion of chlorogenic acid within the plant itself. The results of this research reveal that chlorogenic acid exhibits a massive potential in controlling pest attacks against Solanum melongena L. and can be used as a potential bio-pesticide.

Currently, there is a demand for rapid, sensitive, low-cost, portable, and visualized testing technologies for point-of-care testing (POCT). However, most traditional testing methods face challenges such as long testing times, complicated operations, and high costs, limiting their implementation in resource-limited areas and hindering the fulfillment of POCT demands. Lateral flow assay (LFA) has emerged as an ideal detection technique for POCT, particularly when utilizing gold nanoparticles (AuNPs) as labels. This approach not only enables visualization with the naked eye but also reduces the need for expensive reading instruments. The technologies reviewed in this paper encompass integrated detection technology utilizing amplification technique and LFA, integrated detection technology utilizing clustered regularly interspaced short palindromic repeats (CRISPR) system and LFA, the utilization of surface-enhanced Raman spectroscopy (SERS) in LFA detection technique, the utilization of aptamers in LFA detection technique, and the utilization of DNA barcodes in LFA detection technique. By integrating these advanced techniques, there is significant potential to overcome the limitations of LFA, including low sensitivity, poor specificity, inability to quantify, and false positives, thereby enabling broader applications in resource-constrained settings. Additionally, this article comprehensively evaluates the strengths and weaknesses of each approach, underscoring the immense developmental potential of AuNPs-based LFA in point-of-care testing (POCT).

Regular long-term exercise can benefit the body and reduce the risk of several diseases, such as cardiovascular disease, diabetes, and obesity. However, the proteomic and metabolomic changes, as well as the physiological responses associated with long-term exercise, remain incompletely understood. To investigate the effects of long-term exercise on the human body, 14 subjects with long-term exercise habits and 10 subjects without exercise habits were selected for this study. Morning urine samples were collected and analyzed for untargeted metabolomics and proteomics using liquid chromatography-mass spectrometry. A total of 404 differential metabolites and 394 differential proteins were screened in this research, and the analysis results indicated that long-term exercise may affect energy metabolism, amino acid synthesis and metabolism, nucleotide metabolism, steroid hormone biosynthesis, and the inflammatory response. These findings offer a more comprehensive understanding of the molecular effects of long-term exercise on the human body and provide a basis for future research exploring the underlying mechanisms.

Alkaliptosis, crucial in various cancers, is a specific form of cell death. This study aims to screen a prognosis-related gene in kidney renal clear cell carcinoma (KIRC) using the alkaliptosis gene set and to investigate the roles of the gene in KIRC and its association with alkaliptosis. Transcriptome and clinical information of KIRC patients were collected from the TCGA-KIRC and GSE29609 database. We detected ZACN levels in normal kidney cells and KIRC cells and assessed the ZACN and JTC801-induced alkaliptosis relationship using immunoblotting and pH measurement. In the alkaliptosis gene set (IKBKB, NFKB1, CA9, CHUK, IKBKG, and RELA), NFKB1, CHUK, and IKBKG exhibited differential expression in TCGA-KIRC. Based on these three genes, two alkaliptosis patterns were identified in TCGA-KIRC. The independent prognostic gene for KIRC, ZACN, was screened. High ZACN in KIRC patients indicated a poor prognosis. ZACN was inversely related to the infiltration of anti-cancer immune cells such as CD4 + T cells, macrophages, and neutrophils, and it regulated the immune checkpoint and gene mutations. Patients characterized by high ZACN levels exhibited a heightened drug sensitivity to ABT737_1910, 5-Fluorouracil_1073, etc. ZACN expression in KIRC cells was increased relative to normal kidney cells and was inhibited in a concentration-dependent manner by JTC801. ZACN overexpression suppressed p-p65/p65 expression, increased expression of CA9, and lowered intracellular pH. In KIRC, ZACN inhibits JTC801-induced alkaliptosis. This study sheds light on a novel mechanism involving ZACN and alkaliptosis in KIRC, offering a promising avenue for further research and potential therapeutic interventions in KIRC management.

Endophytic fungi represent a significant renewable resource for the discovery of pharmaceutically important compounds, offering substantial potential for new drug development. Their ability to address the growing issue of drug resistance has drawn attention from researchers seeking novel, nature-derived lead molecules that can be produced on a large scale to meet global demand. Recent advancements in genomics, metabolomics, bioinformatics, and improved cultivation techniques have significantly aided the identification and characterization of fungal endophytes and their metabolites. Current estimates suggest there are approximately 1.20 million fungal endophytes globally, yet only around 16% (190,000) have been identified and studied in detail. This underscores the vast untapped potential of fungal endophytes in pharmaceutical research. Research has increasingly focused on the transformation of bioactive compounds by fungal endophytes through chemical and enzymatic processes. A notable example is the anthraquinone derivative 6-O-methylalaternin, whose cytotoxic potential is enhanced by the addition of a hydroxyl group, sharing structural similarities with its parent compound macrosporin. These structure-bioactivity studies open up new avenues for developing safer and more effective therapeutic agents by synthesizing targeted derivatives. Despite the immense promise, challenges remain, particularly in the large-scale cultivation of fungal endophytes and in understanding the complexities of their biosynthetic pathways. Additionally, the genetic manipulation of endophytes for optimized metabolite production is still in its infancy. Future research should aim to overcome these limitations by focusing on more efficient cultivation methods and deeper exploration of fungal endophytes' genetic and metabolic capabilities to fully harness their therapeutic potential.

Copper-based nanozyme has shown the superior in the oxidase-like activities due to its electron transfer ability between the Cu(I) and Cu(II) sites during the catalytic reactions. Herein, a Cu(I)-MOF (Cu-Mel) was readily synthesized by a traditional hydrothermal process using the precursors of Cu⁺ and melamine, which was then used in the laccase-like catalytic reactions for the first time. Some means, such as X-ray diffraction (XRD), Scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FT-IR), and X-ray photoelectron spectroscopy (XPS), were employed to character the microstructure of the Cu-Mel. The catalytic oxidation of the 4-aminoantipyrine (4-AP) and 2,4-dichlorophenol (2,4-DP) was adopted to evaluate the laccase-like catalytic ability of the resulting Cu-Mel. The catalytic conditions including the temperatures, the presence of alcohols, and the ionic concentrations were varied to optimize the laccase-like activities, based on that, the highest laccase-like catalytic activity is presented with a higher maximum reaction rate (V_max). The good storage stability is also presented by the Cu-Mel. The Cu-Mel was utilized in the degradation of Congo red, showing a good degradation efficiency. These findings facilitate the development of the laccase mimics and serve as a foundation for the design and applications of Cu-MOFs in the nanozyme realm.