Standard reagents and solvents were commercially available and used without further purification, unless otherwise noted: sodium chloride, 0.9% USP and sterile water for Injection, USP were purchased from Hospira; Dehydrated Alcohol for Injection, USP was obtained from Akorn Inc.; Ascorbic Acid for Injection, USP was acquired from Bioniche Pharma; molecular sieves were purchased from Alltech; and HPLC columns were acquired from Phenomenex. Other synthesis components were obtained as follows: sterile filters were acquired from Millipore; C18 Sep-Paks were purchased from Waters Corporation; 10 cc sterile vials were obtained from HollisterStier. Sep-Paks were flushed with 10 mL of ethanol followed by 10 mL of sterile water prior to use. [11C]CO2 was produced with a General Electric (GE) PETTrace cyclotron. High purity nitrogen containing 0.5% O2 was irradiated with 16.5 MeV to generate [11C]CO2, which was delivered to a GE TRACERlab FXC-Pro synthesis module via stainless steel lines. Radiochemical syntheses were carried out using GE TRACERLab FXC-Pro synthesis modules housed in Comecer hot-cells. Doses produced were assessed via standard quality control techniques (see below) and were appropriate for rodent studies.
Sterile filter
Manufactured by Merck Group
Sourced in United States, Germany, Italy
Sterile filters are laboratory equipment designed to remove particulate matter and microorganisms from fluids, gases, or other substances. They function by utilizing a porous membrane that traps contaminants while allowing the desired components to pass through.
Automatically generated - may contain errors
Lab products found in correlation
Sep pak c18, by Waters Corporation (5 mentions)
Hplc grade acetonitrile, by Thermo Fisher Scientific (5 mentions)
Lc 2010a ht system, by Bioscan (3 mentions)
Dehydrated alcohol for injection, by Akorn (2 mentions)
Porapak q, by Waters Corporation (2 mentions)
Qma light and c18 light sep paks, by Waters Corporation (2 mentions)
Sterile water for injection, by Pfizer (2 mentions)
Lysis buffer, by Merck Group (2 mentions)
Rpmi 1640, by Merck Group (2 mentions)
Pettrace cyclotron, by GE Healthcare (1 mentions)
Tracerlab fxc pro synthesis module, by GE Healthcare (1 mentions)
Decitabine, by Merck Group (1 mentions)
5 aza 2 deoxycytidine decitabine, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
B fc 1000 radiation, by Bioscan (1 mentions)
Solvents and reagents, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Sodium chloride (nacl), by Pfizer (1 mentions)
B fc 1000, by Bioscan (1 mentions)
Sep pak, by Waters Corporation (1 mentions)
29 protocols using sterile filter
1
Radiolabeled Compound Production
2
Aflatoxin B1 Degradation by Strain N17-1
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The effects of cell free supernatant, bacterial cells and intracellular cell extracts of strain N17-1 on reduction of AFB1 were studied according to method as described previously [16 (link),22 (link),36 (link)]. Strain N17-1 was pre-cultivated in 4 mL NB at 37 °C for 12 h agitation at 180 rpm, and 1 mL of the culture was transferred to 100 mL of the same medium. After cultivation at 37 °C with shaking at 180 rpm for 48 h, the cells were harvested by centrifugation (5000× g, 10 min, 4 °C). The supernatant was collected and tested for AFB1 degradation.
The pellets were washed twice with phosphate buffer (50 mM, pH 7.0) before re-suspended in the phosphate buffer. The AFB1 degradation analysis was performed as described above. The phosphate buffer substituted bacterial cell suspensions in control.
Pellets were suspended in phosphate buffer (50 mM, pH 7.0; 3 mL buffer per gram cell mass). The suspension was disintegrated by using ultrasonic cell disintegrator (Ningbo Xinzhi Instruments Inc., Ningbo, China). The suspension was centrifuged at 12,000 g for 10 min at 4 °C. The supernatant was filtered aseptically using sterile filters of 0.22 μm pore size (Millipore, Darmstadt, Germany). The AFB1 degradation tests were performed as described above. Phosphate buffer substituted intracellular cell extracts in control.
The pellets were washed twice with phosphate buffer (50 mM, pH 7.0) before re-suspended in the phosphate buffer. The AFB1 degradation analysis was performed as described above. The phosphate buffer substituted bacterial cell suspensions in control.
Pellets were suspended in phosphate buffer (50 mM, pH 7.0; 3 mL buffer per gram cell mass). The suspension was disintegrated by using ultrasonic cell disintegrator (Ningbo Xinzhi Instruments Inc., Ningbo, China). The suspension was centrifuged at 12,000 g for 10 min at 4 °C. The supernatant was filtered aseptically using sterile filters of 0.22 μm pore size (Millipore, Darmstadt, Germany). The AFB1 degradation tests were performed as described above. Phosphate buffer substituted intracellular cell extracts in control.
3
Ethanolic Extraction of Propolis and Bee Pollen
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The procedure described by Dziedzic et al. (2013) [20 (link)] with modifications was used to prepare an ethanolic extract of propolis and ethanolic extract bee pollen. Crushed propolis (40g) or bee pollen (40 g) (apiary, Poland) was mixed with 96.0% ethanol (100ml). The suspensions were stored in the darkness at room temperature (25°C) with shaking (200 rpm, 6 hours per day) for 4 days. Then the infusion was placed at -20°C for precipitating alcohol insoluble compounds of propolis. After 24 hours, the resulting infusion was filtrated through sterile filters (0.45 μm, Millipore). The 40.0% EEP and 40.0% EEBP were stored at room temperature (25°C).
4
Oligo-fucoidan Extraction and Characterization
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The fucoidan powder from Laminaria japonica, a commercial product named Hi-Q Oligo-fucoidan®, was provided by Hi-Q Marine Biotech International Ltd. (New Taipei City, Taiwan). Briefly, the crude extract of Laminaria japonica was eluted with a NaCl gradient on a DEAE (Diethylaminoethyl)-Sephadex A-25 column. The fucose- and sulfate-enriched fraction eluted at a higher concentration of NaCl was collected and then hydrolyzed with glycolytic enzyme preparation to obtain our oligo-fucoidan (OF) sample [54 (link)]. The characteristics of oligo-fucoidan (OF) were as follows: average molecular weight of <667 Da with a 85.9% fucose content (127.2 ± 1.3 μmol/g), sulfate content 28.4% ± 2.1% (w/w), protein content 4.3% ± 0.3% (w/w), fat content 0.6% ± 0.1% (w/w), ash 4.1% ± 0.1% (w/w) and moisture content 3.9% ± 0.8% (w/w) [29 (link)]. It was dissolved in double-distilled H2O and stirred at 25 °C for 30 min. The dissolved solution was filtered using 0.22 μm sterile filters (Millipore, Billerica, MA, USA). decitabine (5-aza-2′-deoxycytidine, Cat #A3656, Sigma-Aldrich, St Louis, MO, USA) was used as a demethylating agent. Stock solutions of decitabine (20 mM) were dissolved in dimethyl sulfoxide (Cat #D2650, Sigma-Aldrich).
5
Preparation of Sterile Pharmaceutical Solutions
6
Radiolabeled Compound Synthesis and Purification
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Reagents and solvents were purchased from Aldrich Chemical or Fisher Scientific and were used without further purification unless noted. Chromatography columns for HPLC analysis and purification were purchased from Phenomenex or Waters. High performance liquid chromatography (HPLC) was performed using a Shimadzu LC-2010A HT system equipped with a Bioscan B- FC-1000 radiation detector. Sodium chloride, 0.9% USP and sterile water for Injection, USP were purchased from Hospira; Dehydrated Alcohol for Injection, USP was obtained from Akorn Inc. Sterile filters were acquired from Millipore; 10 cc sterile vials were obtained from Hollister-Stier; C18 sep-paks were purchased from Waters Corporation and were flushed with 10 ml of ethanol followed by 10 ml of sterile water prior to use. BCPP-EF reference standard 1 and labeling precursor 15 were synthesized in house. Detailed procedures are provided in the Supplementary Material .
7
Cytotoxicity Evaluation of Dental Materials
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Test materials used in this study were ProRoot MTA white-colored formula (Dentsply Tulsa Dental, Tulsa, Oklahoma, USA) and NFC (Sanat Avaran Vista, Iran).
After sterilization with an ultraviolet beam, the materials were mixed according to the manufacturer's guidelines under aseptic conditions.
Freshly mixed materials were placed into the plastic molds with 5-mm diameter and 3-mm height, and then incubated at 37˚C and 95% humidity for 24 hours.
After complete setting, extracts of the materials were produced as follows: 5 ml of complete Dulbecco's Modified Eagle Medium
(DMEM; Sigma Chemical Co., St. Louis, MO, USA) containing 10% Fetal Bovine Serum (FBS; Gibco, Grand Island, NY, USA) was mixed with 1 mg of each test
material in sterile vials. The mixture was incubated at 37˚C at 100% relative humidity for 72 hours. The medium was then drawn off and
filtered using sterile filters at 0.22 µm (Millipore; Billerica, MA, USA). The extracts were serially diluted 1:2 with complete DMEM to
achieve a total of six concentrations of each extract (neat, 1/2, 1/4, 1/8, 1/16, 1/32).
The cells in DMEM served as the negative control group. DMEM alone, in 96 well tissue culture plates, were used as the positive control group.
After sterilization with an ultraviolet beam, the materials were mixed according to the manufacturer's guidelines under aseptic conditions.
Freshly mixed materials were placed into the plastic molds with 5-mm diameter and 3-mm height, and then incubated at 37˚C and 95% humidity for 24 hours.
After complete setting, extracts of the materials were produced as follows: 5 ml of complete Dulbecco's Modified Eagle Medium
(DMEM; Sigma Chemical Co., St. Louis, MO, USA) containing 10% Fetal Bovine Serum (FBS; Gibco, Grand Island, NY, USA) was mixed with 1 mg of each test
material in sterile vials. The mixture was incubated at 37˚C at 100% relative humidity for 72 hours. The medium was then drawn off and
filtered using sterile filters at 0.22 µm (Millipore; Billerica, MA, USA). The extracts were serially diluted 1:2 with complete DMEM to
achieve a total of six concentrations of each extract (neat, 1/2, 1/4, 1/8, 1/16, 1/32).
The cells in DMEM served as the negative control group. DMEM alone, in 96 well tissue culture plates, were used as the positive control group.
8
Purification and Characterization of Reagents
9
Preparation and Purification of Radiopharmaceuticals
10
Radiolabeled Compound Purification Protocol
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Unless otherwise stated, reagents and solvents were commercially available and used without further purification: sodium chloride, 0.9% USP, and sterile water for injection, USP, were purchased from Hospira; ethanol was purchased from American Regent; HPLC grade acetonitrile was purchased from Fisher Scientific. Other synthesis components were obtained as follows: sterile filters were obtained from Millipore; sterile product vials were purchased from Hollister-Stier; C18 Sep-Paks were purchased from Waters Corporation. C18 Sep-Paks were flushed with 10 mL of ethanol followed by 10 mL of water prior to use. Radio-HPLC was performed using a Shimadzu LC-2010A HT system equipped with a Bioscan B-FC-1000 radiation detector.
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