Spontaneously immortalized microglia, SIM-A9 cell line (cat.#CVCL_5I31) was purchased from Kerafast (Boston, MA) and U-87 MG (ATCC HTB-14) cell line was purchased from ATCC (Manassas, VA). SIM-A9 cells were maintained in complete growth media containing Dulbecco’s Modified Eagle Medium/F12 (DMEM/F12, HyClone, Logan, UT) supplemented with L-glutamine, sodium pyruvate, heat-inactivated fetal bovine serum; 10% (FBS, HyClone) and horse serum; 5% (HS, Gibco Molecular Probes, New Zealand). The media were protected against microbial contamination using 1.5 μg/mL penicillin and 1.5 U/mL streptomycin (Pen Strep, Gibco Life Technologies Corp., Grand Island, NY). The cells were incubated in a humidified 5% CO2 incubator at 37±0.5°C (Isotemp, Thermo Fisher Scientific). Prior to passaging, the cells were washed using 1x Modified Dulbecco’s Phosphate Buffered Saline; DPBS (HyClone) and were dissociated using 1x enzyme-free Cell Dissociation Buffer containing 1mM EGTA (EMD Millipore Corp, MA), 1mM EDTA (Fisher Bioreagents, Darmstadt, Germany) and 1mg/mL dextrose in DPBS.
Isotemp
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Isotemp is a laboratory incubator used for maintaining a constant temperature environment. It is designed to provide precise temperature control for various applications in the laboratory setting.
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Lab products found in correlation
Whatman, by Cytiva (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Coreldraw x4, by Corel (2 mentions)
Ethylene diamine tetraacetic acid (edta), by Thermo Fisher Scientific (1 mentions)
Ethylene glycol tetraacetic acid (egta), by Merck Group (1 mentions)
Tryple express enzyme, by Thermo Fisher Scientific (1 mentions)
U 138 mg atcc htb 16, by American Type Culture Collection (1 mentions)
U 87 mg atcc htb 14, by American Type Culture Collection (1 mentions)
Hydrochloric acid (hcl), by Thermo Fisher Scientific (1 mentions)
Concentrated nitric acid, by Thermo Fisher Scientific (1 mentions)
Microplate reader, by Bio-Rad (1 mentions)
Goldfisch fat extractor, by Labconco (1 mentions)
B10p benchtop meter, by Avantor (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Neurobasal medium, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Merck Group (1 mentions)
Cytosine arabinoside, by Merck Group (1 mentions)
Sonic dismembrator ultrasonic processor, by Thermo Fisher Scientific (1 mentions)
Vcx 1500, by Sonics (1 mentions)
Calcium nitrate tetrahydrate, by Loba Chemie (1 mentions)
45 protocols using isotemp
1
Culturing Spontaneously Immortalized Microglia
2
Cell Culture Protocol for U-87 MG and U-138 MG
3
Determination of Lead in Kidney Tissue
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Pb concentrations in kidney homogenate were analyzed by flame atomic absorption spectroscopy (FAAS). Processing of the samples was performed as previously described [14 (link)]. The frozen kidney homogenate samples were dried at 65°C for 24 h. Dried samples were then weighed and ashed at 500°C for 12~16 h in a muffle furnace (Isotemp, Thermo Fisher Scientific, ON, USA). Ashed samples were digested by heating to a slow boil in 1 mL of concentrated hydrochloric acid (10 M, Thermo Fisher Scientific) and 3 mL of concentrated nitric acid (6 M, Thermo Fisher Scientific) over 4~6 h. One to two drops of hydrogen peroxide (H2O2, 4% w/v) were then added, and samples were boiled until only 0.5 mL of liquid remained. Water was added to concentrated samples (15~20 mL), and absorbance readings were performed at 283.3 nm using FAAS (Spectra AA-10 Varian Spectrophotometer). Standard curves for Pb were prepared from commercially available standards (Sigma). BSA 1577A and Tort-2 (National Institute of Standards and Technology, MD, USA) were used as controls for tissue digestion and preparation procedures.
4
Evaluating Adipose-Derived Stem Cell Viability
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The hADSCs were cultured in CEFOgroTM ADMSC Human Adipose tissue-derived MSC Growth Medium containing CEFOgroTM ADMSC Supplement (10% FBS, 0.02% Penicillin & Streptomycin) (growth medium (GM)) or DMEM containing 10% FBS, 1% PS, 10 mM β-glycerol phosphate disodium salt hydrate, 300 µM ascorbic acid, and 0.1 µM dexamethasone (osteogenic medium (OM)) in a 5% CO2/37°C incubator (Isotemp, Thermo Fisher Scientific). The GM and OM were replaced every 3 days in all cell experiments using hADSCs. The hADSCs were dived into groups including non-treated (control), 20 µM GNPs treated, 20 µM SPVD treated, and 20 µM VGNPs treated groups. Each group was seeded onto a 24-well culture plate at a seeding density of cells/well (n = 4) including GM. EZ-Cytox was used to evaluate optical densities of hADSCs. At 1, 3, and 7 days of evaluations, hADSCs seeded in 24-well culture plates were washed by DPBS and were treated EZ-Cytox solution. This was diluted using free GM including no supplements at a ratio of 1:10. After 2 h of incubation, the intensity was measured by a microplate reader (BioRad, Hercules, USA) at a wavelength of 450 nm.
5
Microwave Plasma Surface Modification of Cotton
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Purified cotton fibers (never ground intact fibers) were oven-dried at 105 °C overnight (Isotemp, Thermofisher Scientific, Pittsburg, PA, USA) and subsequently subjected to microwave oxygen plasma treatment using a plasma chamber (PLASMAtech Inc., Erlanger, KY, USA) (flow rate = 60 mL/min, pressure = 25 Pa, generator frequency = 2.45 GHz, time = 8 min to 48 min with an increment of 4 min, power = 500 W). Plasma surface-modified (PSM) cotton fibers were identified as PSM-0 (untreated), PSM-8 (8 min), PSM-12 (12 min), PSM-16 (16 min), etc.
6
Cell Culture in Laminar Flow Cabinet
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All cell culture work was conducted in a type 2 laminar flow cabinet (ThermoFisher Scientific, 41940037) and all solutions were pre-warmed in a water bath (Isotemp©; ThermoFisher Scientific, 300232999) at 37°C. Cultured cells were incubated at 37°C in a humidified atmosphere with 5% carbon dioxide (CO2) in a CellXpert incubator (Eppendorf, 6734I6002000).
7
Proximate Analysis and pH Monitoring of PPF Fermentation
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Moisture content was determined gravimetrically by calculating the mass loss after drying 2 g sample in an oven (APT.line™ ED, BINDER GmbH, Tuttlingen, Germany) at 130 °C for 1 h, according to AOAC method 925.10 (18 ). Protein content (N/%·6.25) was determined according to AOAC method 920.87 (19 ) using a Kjeldahl micro digestor (model 6030000; Labconco) and distillation unit (Rapid Still I; Labconco). Ash content was determined according to AOAC method 923.03 (20 ) in which 3 g sample was placed overnight in a muffle furnace (Isotemp®; Thermo Fisher Scientific, Waltham, MA, USA) at 550 °C and expressed as the difference of the mass of the sample before and after heating. Crude lipid was determined gravimetrically after solvent extraction with ether according to AOAC method 920.39 (21 ) using a Labconco Goldfisch fat extractor. Proximate analysis was reported on a dry mass basis. To determine the pH, 15 g of PPF fermentation medium at each time interval (0, 1, 5, 9, and 11 h) were transferred to a 25-mL beaker and the pH was measured under stirring conditions using a pH meter (B10P Benchtop Meter; VWR, Mississauga, ON, Canada) and magnetic stirrer plate (RO 5; IKA Works Inc., Wilmington, NC, USA).
8
Osteogenic Differentiation of hASCs
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The hASCs (Cefobio, Seoul, Republic of Korea) were cultured in growth medium (GM) or osteodifferentiation medium (OM) at 37°C in 5% CO2 incubator (Isotemp, Thermo Fisher Scientific). The GM was composed of DMEM, 10% FBS, and 1% PS. The OM was composed of GM, 10 mM β-glycerol phosphate disodium salt hydrate, 300 µM ascorbic acid, and 0.1 µM dexamethasone. In the all cell experiments using hASCs, and the GM and OM were changed every 3 days.
9
Enzymatic Hydrolysis of Rice Protein Isolate
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Hydrolysis of RPI were carried out a in a 200 mL jacketed reactor. Temperature was water-bath controlled (Isotemp, Thermo Fisher Scientific, Waltham, MA, USA) and pH was maintained constant with a 902 Titrando system (Metrohm Ltd., Herisau, Switzerland) with 0.5 mol L−1 HCl. Initial RPI concentration was 1% (w/v, based on protein purity and dry matter basis). The suspension was driven to appropriate conditions (temperature and pH) prior to enzyme addition.
The initial selective hydrolysis on RPI with the Prolyve® enzyme was set at pH 3.0, 50 °C, and E/S ratio of 1/500 (g enzyme/g substrate). The final volume was 50 mL. The reaction was stopped after 7 h of hydrolysis by a pH-shift from 3 to 9.
For the mechanistic study, a suspension of RPI was implemented in a final volume of 60 mL. Temperature and pH range were according to supplier’s data: 40–50–60 °C and pH 3.0–4.0–5.0. The applied E/S ratio range was 1/100–1/500–1/1000 (g enzyme/g substrate). Samples (1 mL) were collected at 0–0.25–0.5–1–3–5 h.
The initial selective hydrolysis on RPI with the Prolyve® enzyme was set at pH 3.0, 50 °C, and E/S ratio of 1/500 (g enzyme/g substrate). The final volume was 50 mL. The reaction was stopped after 7 h of hydrolysis by a pH-shift from 3 to 9.
For the mechanistic study, a suspension of RPI was implemented in a final volume of 60 mL. Temperature and pH range were according to supplier’s data: 40–50–60 °C and pH 3.0–4.0–5.0. The applied E/S ratio range was 1/100–1/500–1/1000 (g enzyme/g substrate). Samples (1 mL) were collected at 0–0.25–0.5–1–3–5 h.
10
Preparation and Characterization of Activated Bauxite
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Bauxite was collected from a mine in Visakhapatnam, Andhra Pradesh, India. Gibbsite was received from Alcoa. After oven drying each sample at 100 °C for 24 h to remove moisture, 5 g of bauxite was milled for 15 min and 5 g of gibbsite for 1 h in a stainless steel milling jar of a shaker ball mill (SPEX 8000 or SPEX 8000M) to generate micron sized powders. The milling time for the gibbsite was chosen from among several possible time intervals because it provided material with the surface area (by BET nitrogen adsorption, Gibbsite: 15.1 ± 2.1 m2/g) that was closest to that previously reported for Bauxite: 11.0 ± 3.0 m2/g (Cherukumilli et al., 2017b ). Some of the powdered bauxite was then heated at 300 °C for 4 h in a muffle furnace (Fisher Scientific, IsoTemp) to produce “thermally activated bauxite” according to the procedure of Cherukumilli et al. (2017b) Activated bauxite was demonstrated in our previous work to have significantly higher adsorption capacity for fluoride as compared to raw bauxite, as well as a higher surface area by BET, Activated Bauxite: 173 ± 25 m2/g (Cherukumilli et al., 2017b ). These values are consistent with trends in surface area of aluminum oxide and hydroxide species reported in the literature (Das et al., 2005 (link); Fleming and Goodboy, 1990 ).
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