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Environmental Compliance and Control for Radiopharmaceutical Production
Published in Michael Ljungberg, Handbook of Nuclear Medicine and Molecular Imaging for Physicists, 2022
Ching-Hung Chiu, Ya-Yao Huang, Wen-Yi Chang, Jacek Koziorowski
Generally, radiopharmaceuticals used in a clinical situation need to be sterile, and the production process involves two parts: aseptic manufacture or on-site preparation, and terminal sterilization. In other words, radiopharmaceutical manufacturing and in-hospital radiopharmaceutical preparation are carried out either in laminar flow cabinets (LFC), lead-shielded workstations (hot cells), or in closed radiological protection workstations in a well-controlled cleanroom, and the aseptic operational process conducted at some or all stages, and terminal sterile filtration of final products [13]. In Europe, all aseptic manufacturing of sterile radiopharmaceuticals has to comply with the requirements described in the current [16] version (noted as EU GMP 2020) [17] of Annex 1 of the Eudralex Volume 4 guide. However, the basic requirements presented in PIC/S Guide PE 010-4 (noted as PE 010-4) [13] cover on-site preparation of sterile radiopharmaceuticals for direct supply to patients in hospital radiopharmacies of the PIC/S member country.
Polar body biopsy and its clinical application
Published in David K. Gardner, Ariel Weissman, Colin M. Howles, Zeev Shoham, Textbook of Assisted Reproductive Techniques, 2017
For aCGH, qPCR, and NGS, the first and second PBs must be transferred separately into individual PCR reaction tubes. This step should be performed in a laminar flow cabinet in order to avoid any contamination of the sample. Transfer can be easily accomplished by a 0.1-2.5-pL micropipette or by a stripping pipette used for oocyte denudation. Taking the PB into the pipette tip must be done under visual control using a stereomicroscope. Release of the PB in the PCR tube is best accomplished if the tube is prefilled with phosphate-buffered saline (PBS), the amount of which depends on the requirements of the subsequent amplification protocol. A usual protocol is 2.5 pL of PBS and, in this case, the tube should be pre-filled with 2.2 pL of PBS, and the transfer of a single PB is done in 0.3 pL of medium. After this, transfer tubes must be kept or stored in an upright position until amplification. Whole-genome amplification, labeling of amplified DNA, hybridization, and subsequent washing are usually performed according to the instructions provided by the manufacturer.
Definition of HLA-Dw Determinants Using Homozygous Typing Cells and the Mixed Lymphocyte Culture
Published in M. Kam, Jeffrey L. Bidwell, Handbook of HLA TYPING TECHNIQUES, 2020
In order to avoid any possible bacterial or fungal contamination of media, blood, or cultures, sterile technique is used at all times in a laminar flow cabinet. In addition, the following equipment is required: Inverted microscope and hemocytometer (Neubauer chamber) for counting cellsA multidispenser, e.g., 2500 μl Hamilton syringe for dispensing cells or disposable tips with any suitable pipetteTemperature-controlled water bathCentrifuges4°C Refrigerator to store mediaCell freezing machine or -70°C freezerLiquid nitrogen (LN2) to store the frozen cellsCO2 humidified incubator at 37°C to maintain the culturesCell harvestery-Irradiation machineLiquid scintillation analyzer or beta counter
Amphoteric Mannan as an Immune Response Modifier. New Model Immunobiologically Active Candida albicans Mannan-Based Formula
Published in Immunological Investigations, 2023
Ema Paulovičová, Lucia Paulovičová, Alžbeta Čížová, Jana Mečárová, Romana Vrzoňová, Pavol Farkaš, Slavomír Bystrický
Stock solutions and dilutions of natural C.albicans CCY 29-3-100 cellular mannan and the formulas M-Z, MUS-Z1, and MUS-Z2 were prepared aseptically using pre-sterilized disposable plastic wares and sterile, apyrogenic aqua pro injectione (Fresenius Kabi Italia S.r.l., Verona, Italy). The solutions were prepared in a laminar flow cabinet and sterilized with 0.2-μm filter (Q-Max®Syringe filter, Frisenette ApS, Knebel, Denmark) before cell exposure. The laminar flow cabinet was sterilized with 70% ethanol p.a. and UV for 30 min prior to all experiments. The stock solutions were controlled with EndoLISA® ELISA-based Endotoxin Detection Assay (Hyglos, Bernried am Starnberger See, Germany) and measured using the Cytation 5 Imager Multi-Mode Reader (BioTek, Winooski, USA) to determine endotoxin-free exposure conditions.
Hepatotoxic effect of tramadol and O-desmethyltramadol in HepG2 cells and potential role of PI3K/AKT/mTOR
Published in Xenobiotica, 2021
Manar A. Helmy, Hussein Abdelaziz Abdalla, Heba Allah Abd El Rahman, Dalia Alsaied Moustafa Ahmed
Several types of cells were used to assess hepatotoxicity with different sensitivity. However, HepG2 cells were ideal for our work due to lacking metabolic enzymatic activity. The idea of the study was to compare the hepatotoxicity of both the parent drug and the metabolite, so any change of tramadol to one of its metabolites within the cell affects the reliability of the results (Narimatsu et al. 2006). Cell culture procedure was performed in a sterile area using a Laminar flow cabinet biosafety class II level (Baker, SG403INT, Sanford, ME, USA). HepG2 cells were suspended in EMEM medium, 1% antibiotic-antimycotic mixture (10,000 U/ml Potassium Penicillin, 10,000µg/ml Streptomycin Sulphate and 25 µg/ml Amphotericin B) and 1% L-glutamine at 1% L-glutamine at 37 °C under 5% CO2 and cultured for 10 days. Cells were seeded at a concentration of 10 × 103 cells/well in fresh complete growth medium in 96-well microtiter plastic plates at 37 °C for 24 h under 5% CO2 using a water-jacketed Carbon dioxide incubator (Sheldon, TC2323, Cornelius, OR, USA). Media was aspirated, fresh medium (without serum) was added, and cells were incubated either alone (negative control) or with different concentrations of tested drugs. DMSO was not used because of its ability to activate the cytochrome metabolizing enzymes in HepG2 cell line (Narimatsu et al. 2006; Choi et al. 2009).
Silicon dioxide and titanium dioxide particles found in human tissues
Published in Nanotoxicology, 2020
Ruud J. B. Peters, Agnes G. Oomen, Greet van Bemmel, Loes van Vliet, Anna K. Undas, Sandra Munniks, Ronald L. A. W. Bleys, Peter C. Tromp, Walter Brand, Martijn van der Lee
Five types of post mortem human samples were analyzed: liver, spleen, kidney, and parts of the jejunum and ileum. These were obtained from 15 subjects (of which only 12 kidneys, jejunum and ileum were obtained). The complete and intact organs and tissue related to the beginning of the jejunum and the end of the ileum parts of the small intestine were obtained from bodies that were donated to the Department of Anatomy of the University Medical Center Utrecht for educational and research purposes. The bodies, 7 men and 8 women who died at the age of 64–98 years, had been fixed in 4% formaldehyde. During sample preparation, each organ was cut into small pieces and ground to a size range of 0.5–1 mm diameter. All sample handling and sample preparation were carried out in a laminar flow cabinet. To determine potential sample contamination, all materials that have been in contact with the organs were collected. The total-Si and -Ti concentrations in these materials or released by these materials were determined. The sum of the blank contributions of each of these materials was equal to (total-Si) or lower than (total-Si) the limit of detection (LOD) of the chemical analysis method for the tissues.