Antibacterial Nanoparticles with Natural Photosensitizers Extracted from Spinach Leaves.
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ABSTRACT:
SUBMITTER: Ludacka P
PROVIDER: S-EPMC8756605 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:In this study, two kinds of novel carbazole-based ethynylpyridine salts: 3,6-bis[2-(1-methylpyridinium)ethynyl]-9-pentyl-carbazole diiodide (BMEPC) and 3,6-bis[2-(1-methylpyridinium)ethynyl]-9-methyl-carbazole diiodide (BMEMC) have been employed as photosensitizers owing to their excellent antibacterial activity. These molecules possess symmetric A-π-D-π-A-type structures, which would bring in the unique optical properties. The inhibition zone measurement of a gradient concentration from 0 to 100 μM showed BMEPC and BMEMC photoinduced antibacterial activity against Escherichia coli. Diameters of zone of inhibition were up to 15 and 14 mm under laser irradiations. Under the exposure of the laser of 442 nm with a power density of 20 mW/cm2, the minimum inhibitory concentrations (MICs) of BMEPC on E. coli were between 3.5 and 6.9 μM and that of BMEMC were between 9.4 and 18.8 μM, respectively. In the dark experiments as a control, the MIC value is between 6.9 and 13.8 μM for BMEPC, whereas it is between 187.5 and 225.0 μM for BMEMC. By the comparison of the MIC values of BMEPC and BMEMC with laser irradiation and in dark, the laser-induced toxicity on bacteria is more evident, though both of the derivatives have dark toxicity. With the laser irradiation duration of 30 s and 10 min for BMEPC and BMEMC, respectively, the survival rate of E. coli approximates zero. An antibacterial mechanism has been proposed based on the electron paramagnetic resonance characterization, which indicates that a nitride radical is generated under laser irradiation. The carbazole-based ethynylpyridine photosensitizers would provide high potential for further applications in photodynamic therapy.
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Project description:Spinach is a useful source of dietary vitamins and mineral elements. Breeding new spinach cultivars with high nutritional value is one of the main goals in spinach breeding programs worldwide, and identification of single nucleotide polymorphism (SNP) markers for mineral element concentrations is necessary to support spinach molecular breeding. The purpose of this study was to conduct a genome-wide association study (GWAS) and to identify SNP markers associated with mineral elements in the USDA-GRIN spinach germplasm collection.A total of 14 mineral elements: boron (B), calcium (Ca), cobalt (Co), copper (Cu), iron (Fe), potassium (K), magnesium (Mg), manganese (Mn), molybdenum (Mo), sodium (Na), nickel (Ni), phosphorus (P), sulfur (S), and zinc (Zn) were evaluated in 292 spinach accessions originally collected from 29 countries. Significant genetic variations were found among the tested genotypes as evidenced by the 2 to 42 times difference in mineral concentrations. A total of 2402 SNPs identified from genotyping by sequencing (GBS) approach were used for genetic diversity and GWAS. Six statistical methods were used for association analysis. Forty-five SNP markers were identified to be strongly associated with the concentrations of 13 mineral elements. Only two weakly associated SNP markers were associated with K concentration. Co-localized SNPs for different elemental concentrations were discovered in this research. Three SNP markers, AYZV02017731_40, AYZV02094133_57, and AYZV02281036_185 were identified to be associated with concentrations of four mineral components, Co, Mn, S, and Zn. There is a high validating correlation coefficient with r?>?0.7 among concentrations of the four elements. Thirty-one spinach accessions, which rank in the top three highest concentrations in each of the 14 mineral elements, were identified as potential parents for spinach breeding programs in the future.The 45 SNP markers strongly associated with the concentrations of the 13 mineral elements: B, Ca, Co, Cu, Fe, Mg, Mn, Mo, Na, Ni, P, S, and Zn could be used in breeding programs to improve the nutritional quality of spinach through marker-assisted selection (MAS). The 31 spinach accessions with high concentrations of one to several mineral elements can be used as potential parents for spinach breeding programs.
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Project description:Minimally processed ready-to-eat (RTE) vegetables are increasingly consumed for their health benefits. However, they also pose a risk of being ingested with food-borne pathogens. The present study investigated the ability of RTE spinach and rocket to support the growth of Listeria monocytogenes as previous studies provided contradicting evidence. Findings were compared to growth on iceberg lettuce that has repeatedly been shown to support growth. Products were inoculated with a three-strain mix of L. monocytogenes at 10 and 100 cfu g-1 and stored in modified atmosphere (4 kPa O2, 8 kPa CO2) at 8 °C over 7-9 days. Spinach demonstrated the highest growth potential rate of 2 to 3 log10 cfu g-1 over a 9-day period with only marginal deterioration in its visual appearance. Growth potential on rocket was around 2 log10 cfu g-1 over 9 days with considerable deterioration in visual appearance. Growth potential of iceberg lettuce was similar to that of rocket over a 7-day period. Growth curves fitted closely to a linear growth model, indicating none to limited restrictions of growth over the duration of storage. The high growth potentials of L. monocytogenes on spinach alongside the limited visual deterioration highlight the potential risks of consuming this raw RTE food product when contaminated.
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Project description:Chlorophylls and their derivatives are currently used in a wide range of applications. To replace the volatile organic solvents commonly applied for their extraction from biomass, aqueous solutions of non-ionic surfactants are studied herein in the extraction of chlorophylls from spinach leaves. Aqueous solutions of several surfactants were screened, demonstrating that their hydrophilic-lipophilic balance (HLB) plays the pivotal role on the extraction performance, with the best results obtained for surfactants with a HLB ranging between 10 and 13. A response surface methodology (RSM) was then used to optimize operational conditions (surfactant concentration, solid-liquid ratio and temperature), leading to a maximum extraction yield of chlorophylls of 0.94 mg/g. After the extraction step, the chlorophylls-rich extract was concentrated by heating above the surfactant-water cloud point, leading to the separation into two-phases, and to a concentration factor of 9 and a recovery of 97% of chlorophylls in the surfactant-rich phase. The antioxidant activity of the extracts was finally appraised, showing that the antioxidant activity of the aqueous chlorophylls-rich extracts is higher than that obtained with volatile organic solvents. The obtained results show the potential of aqueous solutions of non-ionic surfactants to extract highly hydrophobic compounds from biomass and their possible direct use in cosmetic and nutraceutical applications, without requiring an additional recovery or purification step.
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Project description:We have previously shown that stromal and thylakoid-bound ascorbate peroxidase (APX) isoenzymes of spinach chloroplasts arise from a common pre-mRNA by alternative splicing in the C-terminus of the isoenzymes [Ishikawa, Yoshimura, Tamoi, Takeda and Shigeoka (1997) Biochem. J. 328, 795-800]. To explore the production of mature, functional mRNA encoding chloroplast APX isoenzymes, reverse transcriptase-mediated PCR and S1 nuclease protection analysis were performed with poly(A)+ RNA or polysomal RNA from spinach leaves. As a result, four mRNA variants, one form of thylakoid-bound APX (tAPX-I) and three forms of stromal APX (sAPX-I, sAPX-II and sAPX-III), were identified. The sAPX-I and sAPX-III mRNA species were generated through the excision of intron 11; they encoded the previously identified sAPX protein. Interestingly, the sAPX-II mRNA was generated by the insertion of intron 11 between exons 11 and 12. The use of this insertional sequence was in frame with the coding sequence and would lead to the production of a novel isoenzyme containing a C-terminus in which a seven-residue sequence replaced the last residue of the previously identified sAPX. The recombinant novel enzyme expressed in Escherichia coli showed the same enzymic properties (except for molecular mass) as the recombinant sAPX from the previously identified sAPX-I mRNA, suggesting that the protein translated from the sAPX-II mRNA is functional as a soluble APX in vivo. The S1 nuclease protection analysis showed that the expression levels of mRNA variants for sAPX and tAPX isoenzymes are in nearly equal quantities throughout the spinach leaves grown under normal conditions. The present results demonstrate that the expression of chloroplast APX isoenzymes is regulated by a differential splicing efficiency that is dependent on the 3'-terminal processing of ApxII, the gene encoding the chloroplast APX isoenzymes.
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