Appropriate concentrations of EOs determined based on preliminary tests. 0.1, 0.2, 0.42, 0.87, and 1.8 µl/l air for fennel, 0.001, 0.003, 0.012, 0.042, and 0.15 µl/l air for lavender, and 0.1, 0.18, 0.34, 0.64, and 1.2 μl/l air for tarragon were prepared for concentration-mortality response tests. Filter papers with a diameter of 2 cm were attached to the inner surface of the vial caps with a volume of 50 ml. Desired concentrations of EOs were poured on each paper using a micropipette. Each concentration was replicated four times, and pure acetone (Merck, Germany) was used as a control. Twenty adult insects were placed in each vial, and covered using a net. Then the cap of the vials was screwed tightly and samples were kept at 28 ± 2°C under a relative humidity of 60 ± 5% and a photoperiod of 16:8 h (L: D). After 24 h, the number of dead insects was recorded.
Pure acetone
Manufactured by Merck Group
Sourced in Germany, United States
Pure acetone is a colorless, volatile, and flammable liquid chemical compound. It has a characteristic pungent odor. The core function of pure acetone is as a solvent, used for dissolving a wide range of organic compounds, including fats, oils, paints, inks, and adhesives.
Automatically generated - may contain errors
Lab products found in correlation
Phosphatidylcholine, by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Cholesterol, by Merck Group (1 mentions)
Sodium taurocholate, by Merck Group (1 mentions)
Sodium phosphate, by Merck Group (1 mentions)
Luteolin, by Extrasynthese (1 mentions)
Formic acid, by Merck Group (1 mentions)
Acetic anhydride, by ITW Reagents (1 mentions)
Hipersolv chromanorm, by Avantor (1 mentions)
Luteolin 6 c glucoside, by Extrasynthese (1 mentions)
Methanol hipersolv, by Avantor (1 mentions)
Luteolin 7 o glucoside, by Extrasynthese (1 mentions)
Ethanol, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Milli q water purification system, by Merck Group (1 mentions)
Epigallocatechin gallate (egcg), by Merck Group (1 mentions)
Folin ciocalteu s reagent, by Merck Group (1 mentions)
Formic acid, by Biosolve (1 mentions)
Gallic acid, by Merck Group (1 mentions)
Superfrost plus slide, by Thermo Fisher Scientific (1 mentions)
8 protocols using pure acetone
1
Concentration-Mortality Response of Essential Oils
2
Fractionation and Characterization of Lipids
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Methanol Hipersolv®, absolute ethanol, and pure acetone (Merck®, Darmstadt, Germany) were used for the delipidified infusion fractionation. For the monitorization and characterization of fractions, methanol Lichrosolv and Methanol Hipersolv® (VWR, PA, USA), glacial acetic acid, ethyl acetate, formic acid (98–100%), acetone and toluene (Merck®), deionized water Milli-Q (Millipore Symplicity®, Molsheim, France), diphenylboryloxyethylamine and polyethylene glycol 4000 (Sigma® Chemical Co., St. Louis, MO, USA), p-dimethylaminocynamaldehyde and sulfuric acid (Merck®) were used.
For in vitro evaluation of micellar solubility, sodium taurocholate (European Pharmacopoeia Reference Standard®, Strasbourg, France), phosphatidylcholine (Sigma® Chemical Co., St. Louis, MO, USA), cholesterol (Sigma® Chemical Co., St. Louis, MO, USA), sodium chloride (Merck®), sodium phosphate (Merck®), and chloroform (HiPersolv Chromanorm®, VWR, Radnor, PA, USA) were used. The Liebermann–Buchard reagent was prepared by mixing 0.5 mL of sulfuric acid (J. T. Baker®, Deventer, The Netherlands) in 10 mL of acetic anhydride (Panreac®, Barcelona, Spain), as described by [24 ]. Authentic standards of the flavonoids luteolin (Extrasynthese®, Genay, France), luteolin-6-C-glucoside (isoorientin) (Extrasynthese®), and luteolin-7-O-glucoside (Extrasynthese®) were also used.
For in vitro evaluation of micellar solubility, sodium taurocholate (European Pharmacopoeia Reference Standard®, Strasbourg, France), phosphatidylcholine (Sigma® Chemical Co., St. Louis, MO, USA), cholesterol (Sigma® Chemical Co., St. Louis, MO, USA), sodium chloride (Merck®), sodium phosphate (Merck®), and chloroform (HiPersolv Chromanorm®, VWR, Radnor, PA, USA) were used. The Liebermann–Buchard reagent was prepared by mixing 0.5 mL of sulfuric acid (J. T. Baker®, Deventer, The Netherlands) in 10 mL of acetic anhydride (Panreac®, Barcelona, Spain), as described by [24 ]. Authentic standards of the flavonoids luteolin (Extrasynthese®, Genay, France), luteolin-6-C-glucoside (isoorientin) (Extrasynthese®), and luteolin-7-O-glucoside (Extrasynthese®) were also used.
3
Analyzing Phytochemicals in Dutch Camellia sinensis
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Fresh tea leaves (Camellia sinensis) were collected from tea plants grown in the Netherlands. Immediately following the collection, leaves were transferred to the laboratory and washed with distilled water for further treatment.
Gallic acid and Folin-Ciocalteu’s reagent were purchased from Sigma-Aldrich (St. Louis, MO, USA). Pure acetone, methanol, and ethanol were purchased from Merck (Darmstadt, Germany). Standards of epicatechin (EC), epicatechin gallate (ECG), epigallocatechin (EGC), and epigallocatechin gallate (EGCG) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ultrahigh-performance liquid chromatography/mass spectrometry (UHPLC/MS)-grade acetonitrile with 0.1% (v/v) formic acid and water with 0.1% (v/v) formic acid were purchased from Biosolve (Valkenswaard, The Netherlands). Water was prepared using a Milli-Q water purification system (Millipore, Billerica, MA, USA).
Gallic acid and Folin-Ciocalteu’s reagent were purchased from Sigma-Aldrich (St. Louis, MO, USA). Pure acetone, methanol, and ethanol were purchased from Merck (Darmstadt, Germany). Standards of epicatechin (EC), epicatechin gallate (ECG), epigallocatechin (EGC), and epigallocatechin gallate (EGCG) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ultrahigh-performance liquid chromatography/mass spectrometry (UHPLC/MS)-grade acetonitrile with 0.1% (v/v) formic acid and water with 0.1% (v/v) formic acid were purchased from Biosolve (Valkenswaard, The Netherlands). Water was prepared using a Milli-Q water purification system (Millipore, Billerica, MA, USA).
4
Protein Precipitation and Quantification
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Supernatants were collected after each TNFα treatment and stored at −20 °C. To precipitate secreted proteins, four volumes of pure acetone (Sigma-Aldrich) were added to one volume of supernatant and incubated at −20 °C overnight. After a centrifugation step at 10,000× g for 10 min, supernatant was discarded, and the remaining acetone was air-dried. The pellet was resuspended in RIPA lysis buffer supplemented with anti-proteases and anti-phosphatases. Bradford assay was performed for protein quantification and 50 µg of proteins were used for Western blot analysis. Then, immunoblots were performed as described above. Secreted targets were normalized to BSA.
5
Immunofluorescence Virus Detection Protocol
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Viral antigens were detected by immunofluorescence using specific monoclonal antibodies labelled with fluorescein isothiocyanate (IMAGEN Herpes Simplex Virus (HSV) Type 1 and 2; K610611-2 Thermo Fisher Scientific, Waltham, MA, USA).
After 24 h post-infection, the culture medium was removed and pure acetone (Sigma-Aldrich) was added. After incubating for 20 min to fix the cells, the acetone was removed and allowed to air dry. Monoclonal antibody was added at a 1/2 dilution and incubated at 37 °C for 30 min in an orbital shaker and in a humid chamber. After the incubation time had passed, the excess antibody was removed and two washes were carried out with PBS and one with water for 3 min each. The glass was removed from the Shell-vial, placed on a slide and visualized using the OLYMPUS BX-61 ultraviolet light microscope (Olympus, Tokio, Japan) where the staining of the infected cells wasobserved. In addition, images of different fields of vision were taken and analysed using the free software Image-J with the help of the Technical Scientific Services of the University of Oviedo.
After 24 h post-infection, the culture medium was removed and pure acetone (Sigma-Aldrich) was added. After incubating for 20 min to fix the cells, the acetone was removed and allowed to air dry. Monoclonal antibody was added at a 1/2 dilution and incubated at 37 °C for 30 min in an orbital shaker and in a humid chamber. After the incubation time had passed, the excess antibody was removed and two washes were carried out with PBS and one with water for 3 min each. The glass was removed from the Shell-vial, placed on a slide and visualized using the OLYMPUS BX-61 ultraviolet light microscope (Olympus, Tokio, Japan) where the staining of the infected cells wasobserved. In addition, images of different fields of vision were taken and analysed using the free software Image-J with the help of the Technical Scientific Services of the University of Oviedo.
6
Histological Analysis of RhoA Knockout Murine Kidneys
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Kidneys from RhoAfl/fl or RhoA+/fl (N=3) and RhoAfl/fl Cd21+/Cre (N=6) mice of age 33-49 were embedded in O.C.T.-compound (SAKURA), shock frozen in liquid nitrogen and stored at -80°C. Two µm sections were prepared using a cryo-microtome (Leica cryostat CM1950) with a C35 microtome blade (Feather). Sections were mounted on SuperfrostPLUS slides (Thermo Scientific) and fixed within pure acetone (Sigma Aldrich) for 9 min. Sections were stained with HE stain (Waldeck GmbH & Co.) and PAS stain (Sigma Aldrich). The slides were blinded and the sections were examined independently by two senior pathologists for pathological changes in the general cellularity and architectural features of the kidney.
7
FTIR Analysis of Material Samples
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The spectral measurements were made with a UV-Vis Perkin-Elmer Lambda25 and an FTIR Jasco 6300 spectrometer.
An ATR accessory equipped with a diamond crystal (Pike Technologies, Madison, Wisconsin, USA) allows the collection of FTIR spectra directly on a sample without any special preparation. The FTIR spectra were recorded in the region of 4000–400 cm−1, with a TGS detector, and apodization Cosine. The spectral data were processed with JASCO Spectra Manager software, version 2. Samples were scanned at a 4 cm−1 resolution, accumulation: 100 scans. Background reference spectra were recorded using air after every sample to minimize the interference due to carbon dioxide and water vapor in the atmosphere. Between measurements, the ATR crystal was carefully cleaned using pure acetone (Sigma-Aldrich Co., Saint Louis, MO, USA), then, it was dried with soft tissue [43 (link),44 (link),45 (link)].
All measurements were taken at room temperature (T = 23 °C). For each sample, three replicate spectra were recorded to ensure spectral reproducibility and to assess analytical precision; then, the average spectrum was complete.
An ATR accessory equipped with a diamond crystal (Pike Technologies, Madison, Wisconsin, USA) allows the collection of FTIR spectra directly on a sample without any special preparation. The FTIR spectra were recorded in the region of 4000–400 cm−1, with a TGS detector, and apodization Cosine. The spectral data were processed with JASCO Spectra Manager software, version 2. Samples were scanned at a 4 cm−1 resolution, accumulation: 100 scans. Background reference spectra were recorded using air after every sample to minimize the interference due to carbon dioxide and water vapor in the atmosphere. Between measurements, the ATR crystal was carefully cleaned using pure acetone (Sigma-Aldrich Co., Saint Louis, MO, USA), then, it was dried with soft tissue [43 (link),44 (link),45 (link)].
All measurements were taken at room temperature (T = 23 °C). For each sample, three replicate spectra were recorded to ensure spectral reproducibility and to assess analytical precision; then, the average spectrum was complete.
8
Cellulose Dissolution and Regeneration
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Wood pulp with a degree of polymerization of 450 was provided by Innovia Films Ltd. The ionic liquid 1-ethyl-3-methylimidazolium acetate (EMIMAc) (≥95%), dimethyl sulfoxide (DMSO) (99.9%), pure ethanol, pure acetone and deuterated water (D2O) were purchased from Sigma-Aldrich. Ultra-pure water (H2O) used in the coagulation bath was obtained from a Millipore water system.
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