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42 protocols using up200st

1

Synthesis of Graphene Oxide and Modified Graphene

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An aqueous suspension of GO was prepared using a modified Hummer’s method described in our previous study [49 (link)]. In brief, 0.1 g of flake graphite powder (Sigma Aldrich, St. Louis, MO, USA) was placed in 14 mL of concentrated sulfuric acid (Rushim, Moscow, Russia) with the 0.4 g of potassium permanganate (Rushim, Moscow, Russia). The resulting reaction mixture was placed in a beaker and stirred for three weeks. The reaction was stopped with a 5% solution of hydrogen peroxide (Rushim, Moscow, Russia) (7 mL) and washed with deionized water until the filtrate became neutral. A brown gel-like mass was diluted with water (50 mL) and treated with ultrasound for 5 min on a disperser Hielscher Up 200 St (Hielscher Ultrasonics, Teltow, Germany) at 60 W.
An aqueous suspension of MOG was obtained by the electrochemical exfoliation of graphite. A detailed description of the process was provided in our previous study [50 (link)]. Briefly, a graphite rod (Polyprof-L LLC, Moscow, Russia) was electrochemically exfoliated in an NH4SO3 (Rushim, Moscow, Russia) solution at 10 V. The resulting product was washed with distilled water and dispersed in water by using an ultrasonic disperser Hielscher Up 200 St (Hielscher Ultrasonics, Teltow, Germany) at 30 W for 10 min.
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2

Functional Textile Finishing via Nanoparticle Deposition

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The suspensions were brought to a concentration of 1 mg/mL by diluting previously obtained suspensions. The concentration was determined by weighing the dry residue using Vibra AF-R220CE (Shinko Denshi Co. ltd., Tokyo, Japan). Suspensions of GO/CDs and MOG/CDs were obtained by mixing the suspensions at a ratio of 1:1, followed by ultrasonic treatment at a power of 30 W for 30 min using Hielscher Up 200 St (Hielscher Ultrasonics, Teltow, Germany). Cotton fabrics (bleached calico with a density of 120 g/m2, Ivanovo Garment Factory, Ivanovo, Russia) were used as the base material. Textile samples were dipped into aqueous suspensions of GO, MOG, CDs, GO/CDs, and MOG/CDs at a concentration of 1 mg/mL and treated with mild ultrasound for 30 min at room temperature. The fabrics were then dried under normal conditions until they were completely dry. To evaluate the washing stability, the samples were washed according to GOST 9733.4-83 [51 ]. The samples were placed in a washing solution that consisted of 0.5 g of soap (Belgorod soap facroty, Belgorod, Russia), 0.2 g of soda (Bashkirskaya Sodovaya Kompaniya, Sterlitamak, Russia), and 100 mL of water. The detergent solution in the beaker containing the samples was stirred at 300 rpm using an magnetic stirrer HS Pro-DT (Stegler, Moscow, Russia) at 60 °C for 30 min. Photographs of the fabrics before and after washing are shown in Figure 9.
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3

Preparation of Aqueous Black Pigment Dispersion

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Example 1

The following materials were put into a metal beaker, and manual agitation was performed sufficiently so as to obtain a mixture.

Carbon black CB1: 15 parts by mass

Ion-exchanged water: 85 parts by mass

Basic compound (34-percent potassium hydroxide aqueous solution): 2.2 parts by mass

The resulting mixture was further subjected to ultrasonic dispersion for 3 minutes by using an ultrasonic dispersing machine described below. In this regard, the applied energy per unit carbon black at this time was 0.7 W/g.

Ultrasonic dispersing machine (UP200St produced by hielscher)

Maximum output: 200 W

Frequency: 26 kHz

After a lapse of 30 minutes from finishing of dispersion, 15 parts by mass (3 parts by mass as resin solid content) of Urethane resin aqueous solution PUS was added as a resin having anionic groups, and manual dispersion was performed sufficiently so as to obtain an aqueous black pigment dispersion.

At this time, the proportion of resin in the resulting aqueous black pigment dispersion was 3/15=0.2, where the proportion of resin was denoted as “mass of resin having anionic group/mass of carbon black” and was an indicator of the amount of addition of the resin having anionic groups.

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4

Preparing Preformed Fibrils of α-Synuclein

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Recombinant full-length human WT α-syn with a N-terminal His-tag and a C-terminal Avi-tag was obtained as previously described [34 (link)]. To prepare PFFs, 0.9 mg/mL recombinant protein in PBS was incubated at 37°C with constant agitation for 4 days. The presence of cross-β-sheet structure and final plateau of aggregation was verified by Thioflavin T fluorescence. The reaction product was centrifuged (20000 g) at 4°C for 1 h and the supernatant was discarded. The pellet was resuspended in PBS (1.67 mg/mL) and sonicated with a Hielscher UP200St ultrasonic homogeniser (two cycles of 1 min in pulses of 10 s and 5 s rest). The resulting PFFs were aliquoted, snap frozen and stored at −80°C. Commercial untagged human WT α-syn PFFs (StressMarq) were also employed. Prior to use, PFFs were further diluted in PBS into the required concentration and re-sonicated (1 min in pulses of 10 s and 5 s rest). PFFs were directly added to the medium of WT neurons to reach a final concentration of 5 μg/mL. Equal volume of PBS was added as control condition.
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5

Quantifying Drug Content in Transdermal Patch

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An accurately weighed 2.54 cm 2 (trans)dermal patch was dissolved in 50 mL methanol by mechanically shaking and sonication (UP200st, Hielscher, D). Afterwards, the samples were left to rest overnight and, then, diluted 1:1 with mobile phase described below. Before the injection, samples were filtered with a 0.45 μm polypropylene filter (VWR International, I). The drug content in the (trans)dermal patch was calculated as a function of both the matrix mass (μg/g) and area (μg/cm 2 ).
The results were expressed as the mean ± standard deviation of three specimens for each formulation.
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6

Optimized Ultrasound-Assisted Extraction

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Plant material was ground and then homogenized in a blender (Grindomix GM200, Retsch, Haan, Germany) at 5000 rpm (time: 20 s) and then sieved (150 μm mesh size). Moisture presented a value of about 84.50% performed by a MA Moisture Sartorius analyzer (Goettingen, Germany). All analyses were performed in triplicate and the results expressed as mean values (±standard deviations). PUAE was carried out directly by a sonicator (Hielscher UP200St, Teltow, Germany) composed by a titanium sonotrode (diameter: 7 mm) at a constant frequency of 26 kHz. The experimental conditions (amplitude: 30%; duty cycle: 70%; time of extraction: 20 min) were previously optimised on the same raw materials [4 (link),10 (link)].
The extracts obtained (expressed as US) were filtered by Buchner (Whatman n. 1 paper), centrifuged (3000 rpm; 10 min), and dark stored at 4 °C until analysis.
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7

Preparation and Characterization of O/W Nanoemulsions

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A two-step process was employed to prepare the O/W nanoemulsions (Fig. 4b). All pre-emulsions were prepared by a Polytron PCU-2-110 homogenizer (Brinkmann Ind. Westbury, NY, USA). For the CCRD design, the preparation process is described as follows: HSO was mixed with distilled water in desired concentrations as shown in Table 1 to obtain a 50 ml solution. Then the different concentration of lecithin and poloxamer 188 (shown in Table 1) were added into this solution and homogenized for 10 min at a speed setting of half-maximum (= 5). Then the O/W nanoemulsions were formed by ultrasonicating (UP 200ST, Hielscher Ultrasonics, Teltow, Germany) the pre-emulsions for different time using 50% of full power. For the kinetic analysis employed, 5, 10, 20, 30 and 50 ml pre-emulsions were prepared by using HSO, lecithin and polaxamer 88 in the optimal ratio calculated from the CCRD design, using Polytron homogenizer, as described above. Samples were then ultrasonicated at 25, 50 and 100% power levels for upto 1 h. For verification with olive-oil samples, one other nanoemulsion was prepared using olive oil and tween 80 with the same method. To control the temperature (and thus isolate its’ effects) during the ultrasonication process, all samples were kept in ice-bath. All the nanoemulsions were stored at 4 °C for further tests.
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8

Fullerene Protein Hybrids Synthesis

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The C60@protein hybrids were prepared mixing an excess of fullerene powder with a 0.3 mM solution of each protein (5 mL), with a 2:1 stoichiometry. NaOH and HCl 1M were used to adjust pH of the protein solutions. The heterogeneous mixtures were then sonicated in a vial for 120 min using a probe tip ultrasonicator (Ultrasonic Processor UP200St, Hielscher Ultrasonics GmbH, Teltow, Germany), equipped with a sonotrode S26d7, used at 40% of the maximum amplitude). During the process, the sample was refrigerated with an ice bath. The dark brown turbid mixture obtained after the sonication was centrifuged at 10 kRCF. The resulting supernatant was then collected and characterized.
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9

Surfactant-stabilized SWCNT dispersion

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SWCNT powder was introduced to a 2% aqueous solution of SC in such a way that the final concentration of nanocarbon material and volume of the dispersion were 1 mg/mL and 40 mL, respectively. The mixture was homogenized by ultrasound tip sonicator (Hielscher UP200St, Teltow, Germany) with a constant power input of 50 W for 1 h. The processed dispersion was kept in an ice bath to avoid the thermal desorption of the surfactant from the SWCNT surface during the process. The dispersion was centrifuged (Eppendorf Centrifuge 5804 R, Hamburg, Germany) at a constant temperature (18 °C) for 1 h at 15,314× g to remove the non-individualized SWCNTs and impurities. The upper 80% of supernatant after centrifugation was collected and used for experiments.
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10

Preparation and Characterization of αSyn Fibrils

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The αSyn fibrils were prepared using a single monomer batch which was transferred to different buffer conditions (200 μM final monomer concentration) followed by incubation in the benchtop thermo-shaker with constant agitation (1200 rpm) at 37 °C for 7 days. Fibril samples were frozen and stored at −20 °C. Prior to the experiment, fibrils were thawed and sonicated using an ultrasonic probe (Hielscher UP200St). Sonication was carried out in repeating pulses of 3 second duration with 20% amplitude separated by 12 second pauses for 5 minutes (one minute total sonication time) to prevent sample overheating.
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