For the samples of the elicitor-treated and the nontreated group, only the leaves of the above-ground part of the S. chinensis were collected, foreign substances were removed, and naturally dried, and then ground to 40 mesh was used. Then, preparation of extract experimental samples was 1 g of S. chinensis leaves powder was mixed with 100 mL of 70% ethanol, homogenized for 1 min at 16,000 rpm, and agitated for 24 h. Finally, the sample extracts was filtered and concentrated by utilizing a filter paper No. 41 (Whatman Inc., Piscataway, NJ, USA) and Buchi rotavapor (Buchi Labortechnik AG, Flawil, Switzerland). The extracted extract was freezedried (freeze dryer, FD8518, Ilshinbiobase, Yangju, Korea) to remove moisture, and then stored at a low temperature in a sample storage refrigerator at 4 °C or -20 °C and used for experiments.
Fd8518
Manufactured by IlShinBioBase
Sourced in Japan
The FD8518 is a dual channel fluorescence detector designed for high-performance liquid chromatography (HPLC) applications. It features two independent optical channels for simultaneous detection of different fluorescent compounds. The device employs a xenon flash lamp as the excitation source and utilizes photomultiplier tubes (PMTs) for sensitive fluorescence measurements. Key specifications and capabilities of the FD8518 are provided in the product datasheet.
Automatically generated - may contain errors
Lab products found in correlation
No 41 filter paper, by Cytiva (1 mentions)
Jsm 6490lv, by JEOL (1 mentions)
E 1010, by Hitachi (1 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
Vega 2 lsu, by TESCAN (1 mentions)
Whatman no 1 filter paper, by Cytiva (1 mentions)
Penicillin streptomycin, by Cytiva (1 mentions)
Fetal bovine serum (fbs), by Cytiva (1 mentions)
Dmem high glucose medium, by Cytiva (1 mentions)
12 protocols using fd8518
1
Standardized Extraction of Schisandra chinensis Leaves
2
Visualizing Bacterial Cell Morphology under UA-CS NPs
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The impacts of UA-CS NPs on the cell morphology of bacteria was checked under scanning electron microscopy (SEM) as described earlier with slight modification [84 (link)]. Briefly, the overnight grown cells were diluted (1:100) in their respective growth media and treated with sub-MIC of UA-CS NPs. These cell cultures were incubated at 37 ℃ for 4 h under shaking conditions (200 rpm). UA-CS NPs treated cell culture (1 mL) were placed in the 24-well plate containing nylon membrane (0.5 × 0.5 cm) and incubated for 5 h at 37 ℃. The bacterial cells on the nylon membrane surface were fixed using formaldehyde (2%) and glutaraldehyde (2.5%) at 4 ℃ overnight. The cells had been fixed using the above-fixing agents were washed thrice using PBS; these fixed cells were then further dehydrated using increasing concentrations of ethyl alcohol. These nylon membranes were freeze-dried using a freeze dryer (FD8518, ilShinBiobase Co. Ltd., Dongducheon, Korea). These membranes were directly affixed to SEM stubs and coated with gold for 120s using ion-sputter (E-1010, Hitachi, Japan). Finally, bacterial cell morphologies were visualized by using JSM-6490LV (JEOL, Tokyo, Japan) at the magnification of × 5000 and voltage of 15 kV.
3
Fabrication of Usnic Acid-Chitosan Nanoparticles
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The methodology used for the preparation of UA-CS NPs was a single-step ionotropic gelification method as described earlier with slight modifications [22 (link),39 (link)]. Briefly, low molecular weight chitosan (1.2% w/v) was initially dissolved in 1% acetic acid in a total volume of 100 mL of deionized water. The chitosan solution was continuously agitated at the room temperature for 12 h and added with 0.3 g Tween 60 to obtain a homogenous mixture. The mixture was stirred for 60 min at 50 ℃. The usnic acid (0.5%, w/v) solution prepared in dimethyl sulfoxide was slowly added and allowed to mix by stirring for 2 h at room temperature. Two different concentrations of TPP, which are 0.5% and 1.0%, were used for the preparation of UA-CS NPs. The solution of TPP was prepared in a total of 40 mL deionized water and was slowly added dropwise with continuous stirring for 2 h. The light yellow color mixture was centrifuged (13,000 rpm for 15 min) at 4 ℃. Similarly, CS NPs (unloaded with UA) was also prepared using 0.5% and 1.0% TPP. The obtained pellet was washed repeatedly for three times with deionized water and kept at −70 ℃ in a refrigerator to solidify. Finally, the frozen samples were freeze-dried using a freeze dryer (FD8518, ilShinBiobase Co. Ltd., Dongducheon, Korea).
4
Extraction and Characterization of Phragmitis Rhizoma
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Phragmitis rhizoma was purchased from Kwangmyungdang Medicinal Herbs Co. (Ulsan, Republic of Korea) and identified by Dr. Goya Choi, K-herb Research Center, Korea Institute of Oriental Medicine, Republic of Korea. A voucher specimen (KIOM-CRC-#172) was deposited at the KM Convergence Research Division, Korea Institute of Oriental Medicine. The dried Phragmitis rhizoma (500.0 g) was finely ground using a mechanical pulverizer and then subjected to repeated extraction in boiling distilled water (100 °C, 2 h, 10 L × 2 times,) using a reflux extraction system from KOC Biotech (Daejeon, Republic of Korea). The extract solution was filtered through cotton wool, concentrated in the EYELA rotary evaporator system (Tokyo Rikakikai, Tokyo, Japan) and freeze-dried (FD8518, Ilshin Biobase, Dongducheon, Republic of Korea) to produce a final water extract (EPR, 60.21 g, 12.0%). The extract was stored at 4 °C in a dark plastic bottle with a silica gel desiccant until use.
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Phragmitis rhizoma was purchased from Kwangmyungdang Medicinal Herbs Co. (Ulsan, Republic of Korea) and identified by Dr. Goya Choi, K-herb Research Center, Korea Institute of Oriental Medicine, Republic of Korea. A voucher specimen (KIOM-CRC-#172) was deposited at the KM Convergence Research Division, Korea Institute of Oriental Medicine. The dried Phragmitis rhizoma (500.0 g) was finely ground using a mechanical pulverizer and then subjected to repeated extraction in boiling distilled water (100 °C, 2 h, 10 L × 2 times,) using a reflux extraction system from KOC Biotech (Daejeon, Republic of Korea). The extract solution was filtered through cotton wool, concentrated in the EYELA rotary evaporator system (Tokyo Rikakikai, Tokyo, Japan) and freeze-dried (FD8518, Ilshin Biobase, Dongducheon, Republic of Korea) to produce a final water extract (EPR, 60.21 g, 12.0%). The extract was stored at 4 °C in a dark plastic bottle with a silica gel desiccant until use.
5
Extraction and Characterization of Ficus viridissima Fruit
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The dried fruits of F. viridissima were supplied by Kwangmyungdang Medicinal Herbs Co. (Ulsan, Republic of Korea) and identified by Dr. Goya Choi (Herbal Medicine Research Center, Korea Institute of Oriental Medicine, KIOM). A voucher specimen (KIOM-CRC#518) was stored in the Clinical Medicine Division of KIOM, Republic of Korea. The dried materials (1000 g) were extracted for 3 h, twice in a water refluxed system (10 L) and filtered through a mesh strainer and then cotton wool. The filtrates were further concentrated using a rotary evaporator (N-1200A, Rikakikai, Tokyo, Japan) and dried using a freeze dryer (FD8518, IlshinBioBase, Dongduchun, Republic of Korea). The final aqueous extracts of F. viridissima fruits (EFVF) were homogenized and stored in an air-tight container under desiccated condition. For the in vitro studies, EFVF was dissolved at 4 mg/mL in sterile phosphate-buffered saline (PBS) as a stock solution and stored at −80 °C. For the animal studies, EFVF was suspended in a sterile 0.5% CMC solution right before use.
6
Blending and Roasting Varieties of Green Beans
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The 12 varieties of green beans used in this experiment are shown in Table 1 .
The beans were obtained from a local coffee company: 3 kinds of natural and pulped natural, 6 kinds of washed, 2 kinds of honey, and a kind of glazed green. The blending process varied depending on the number of products randomly blended with green beans. Eleven kinds of BC were prepared after mixing 6 or 7 kinds of green beans. The blended green beans were freeze-dried in a freeze dryer (FD8518, Ilshin Biobase Co., Ltd., Dongducheon, Korea) at a temperature of −30°C and a pressure of 0.8 mmHg for 36 h. The dried samples were roasted at 225°C for 15 min and crushed. The crushed sample was used for further analyses.
The beans were obtained from a local coffee company: 3 kinds of natural and pulped natural, 6 kinds of washed, 2 kinds of honey, and a kind of glazed green. The blending process varied depending on the number of products randomly blended with green beans. Eleven kinds of BC were prepared after mixing 6 or 7 kinds of green beans. The blended green beans were freeze-dried in a freeze dryer (FD8518, Ilshin Biobase Co., Ltd., Dongducheon, Korea) at a temperature of −30°C and a pressure of 0.8 mmHg for 36 h. The dried samples were roasted at 225°C for 15 min and crushed. The crushed sample was used for further analyses.
7
Fucoidan-Mediated Synthesis of Gold Nanoparticles
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The entire procedure employed in synthesizing the fucoidan-gold nanoparticles (Fu–AuNPs) was adapted from the previous reports, with a slight modification [29 (link)] (Figure 1 ). The reaction was started by dissolving 1 mM HAuCl4∙3H2O into the sterile deionized water (200 mL) at 60 °C, followed by a continuous stirring. The solution’s alkaline pH (8.5) was adjusted using 0.1 M NaOH. After 1 h stirring, the solution of fucoidan (5 mg) prepared in deionized water was added dropwise, and the pH of the final solution was adjusted to 8.5–9.0. The formation of the AuNPs from the reaction mixture was carried out by striving continuously for 2 h at 60 °C. The formation of the AuNPs was tracked and monitored from the start of the reaction, by measuring the solution’s UV-visible absorption spectra. The scanning of absorption spectra from 200 to 700 nm was carried out using a microplate reader (BioTek, Winooski, VT, USA). In addition, the visible appearance of a wine-red color was also used as an indication of AuNP synthesis. After verifying the synthesis of the AuNPs by measuring the absorption spectra and appearance of the wine-red color, the solution was frozen at −70 °C. The frozen sample was dried in powder form using a freeze drier (FD8518, ilShinBiobase Co. Ltd., Yangju-si, Republic of Korea).
8
Visualizing NP-Treated Microbial Biofilms
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Each NP-treated microbial cell's biofilm architecture was observed using scanning electron microscopy (SEM) [48 (link)]. A nylon membrane (0.5 × 0.5 cm) was put in a 24-well microplate. Cell culture was deposited on the membrane in a 24-well microplate according to the biofilm assay. Cells were additionally treated with PVD-AgNPs, while some were left untreated for the control group. The plate was incubated for 24 h at 37 °C. Biofilm cells were fixed with formaldehyde (2%) and glutaraldehyde (2.5%) in the well overnight at 4 °C. Each well was washed three times with phosphate-buffered saline (pH: 7.4) to remove unattached cells. Increasing ethanol concentrations were used to eliminate moisture from these attached cells. The membranes were applied to the SEM stub after freeze drying (FD8518, ilShinBiobase Co. Ltd., Korea). Both treated and untreated biofilm cells were seen using VEGA II LSU (TESCAN, Czech) microscopy at 10 kV voltage and 8.00 kx (10 μm) magnification.
9
Bacterial Cell Culture Metabolite Profiling
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Bacterial strain C1 was grown in an MRS medium for 24 h at 37 °C,
and when cell growth reached ∼9 log cfu/mL, the cell cultures
were centrifuged (10 000g for 20 min) at 4
°C. Using a freeze-dryer (FD8518, ilShinBiobase Co. Ltd., Yangju-si,
Korea), the recovered filter-sterilized supernatant was freeze-dried
into powder form. The sterilized supernatant was employed for three
separate purposes: (1) testing antibacterial activity, (2) synthesis
of metallic nanoparticles, and (3) secondary metabolite profiling
from LAB strain C1 cell culture.
and when cell growth reached ∼9 log cfu/mL, the cell cultures
were centrifuged (10 000g for 20 min) at 4
°C. Using a freeze-dryer (FD8518, ilShinBiobase Co. Ltd., Yangju-si,
Korea), the recovered filter-sterilized supernatant was freeze-dried
into powder form. The sterilized supernatant was employed for three
separate purposes: (1) testing antibacterial activity, (2) synthesis
of metallic nanoparticles, and (3) secondary metabolite profiling
from LAB strain C1 cell culture.
10
Extraction and Characterization of Forsythia suspensa
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The dried fruits of F. suspensa were supplied from Kwangmyung-dang Medicinal Herbs Co. (Ulsan, Republic of Korea), and their morphology was carefully validated by Dr. Goya Choi Herbal Research Specialized Center, Korea Institute of Oriental Medicine (KIOM), Republic of Korea. A voucher specimen (KIOM010016) was deposited in the Clinical Medicine Division of KIOM. The herb materials were extracted as previously described [32 (link)]. Briefly, the dried materials (1 kg) were refluxed in distilled water (10 L) for 3 h, twice. The extracts were filtered, concentrated using a rotary evaporator (N-1200A, Tokyo, Japan), freeze dried using a freeze drier (FD8518, IlshinBioBase, Dongduchun, Republic of Korea), and then, homogenized. The final product, EFSF was stored under desiccated condition until use.
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