Irradiation Sample Clauses

Irradiation. Irradiation of components containing lymphocytes is performed in order to render those cells incapable of reproduction and to reduce the likelihood of transfusion associated graft vs. host disease (TA-GVHD). TA-GVHD is a rare but devastating complication of transfusion that has a very high mortality rate and no routinely available effective treatment. Prevention is the best management of TA-GVHD. AABB Standards require irradiated blood be provided to recipients at risk of TA-GVHD, recipients receiving blood products from relatives, and recipients receiving HLA- matched or crossmatched blood components. a. Irradiated cellular blood products should be given to, but not limited, to patients with the following:  Transfusions from blood relatives  HLA-matched transfusions  Granulocyte transfusions  Intrauterine transfusions  Congenital immunodeficiency (ie DiGeorge Syndrome)  BMT and stem cell transplantation  Hodgkin disease  NHL  Therapy with purine analogue drugs

Irradiation. High-energy HZE particle irradiations were performed in Brookhaven, NY at the NASA Space Radiation Laboratory (NSRL). The X- ray (low-LET) exposures were conducted at Emory University using an X-RAD 320 biological irradiator (Precision X-Ray, North Branford, CT).

Irradiation. Exponentially growing cells were seeded in 10-cm dishes or 6-well plates. Twenty-four hours later, the cells were exposed to recombinant viruses AdMLP.apoptin or AdCMV.LacZ at an MOI of 5 at 37ºC. Fresh medium (2 mL) was then added to each well and incubation was continued for 24 hrs. One day after the viral infection, cells were irradiated using a 6 MV linear accelerator. Doses were given in a single fraction of 2–6 Gy. After irradiation, the dishes were immediately returned to the incubator. First treated cells were subcellularly fractionated. The cell monolayer (106 cells/10-cm dish) was rinsed twice with ice-cold PBS, scraped and centrifuged at 2000 g for 5 min. The cells were resuspended in cell extraction buffer (300 mM sucrose, 10 mM Hepes [pH 7.4], 50 mM KCl, 5 mM EGTA, 5 mM MgCl2, 1 mM dithiothreitol, 1 mM phenylmethylsulfonyl fluoride, and 100 µM cytochalasin B), left on ice for 30 min, and homogenized by means of vortexing the lysates. Unbroken cells and nuclei were pelleted by centrifugation for 5 min at 2000 g. Heavy membranes were removed from the resulting supernatant by centrifugation for 5 min at 14000 g. The resulting supernatant is the crude cytosolic fraction. All samples were frozen in liquid nitrogen and stored at -80° C until analysis by sodiumdodecylsulfide-polyacrylamide gel electrophoresis (SDS-PAGE). Next, the cytoplasmic cytochrome c levels were determined. Equal amounts of cytosolic protein extract were boiled in reducing sample buffer for 5 min, loaded in a lane of a sodiumdodecylsulfide-15% polyacrylamide (SDS-PAA) gel, and electroblotted onto Immobilon-P membranes (Millipore, Bedford, MA, USA). The membranes were blocked for 1 hour in PBST-buffer (10 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.05% Tween-20) containing 5% nonfat dried milk. Subsequently, the blots were then incubated during overnight with monoclonal antibody 7H8.2C12 (Pharmingen; dilution 1:1000) to detect cytochrome c. Following overnight incubation, membranes were washed and incubated for 1 h at room temperature with secondary antibody. Membranes were washed three more times and incubated for 5 min at room temperature with Super Signal HRP substrate (Xxxxxx). Positive signals were visualized by enhanced chemiluminescence according to the manufacturer’s protocol (Amersham, Roosendaal, the Netherlands). Cells (8 × 105/dish) were grown in 60-mm plates and treated as follows: Cells were collected by gentle scraping, washed once with ice- cold PBS, and lysed in lysis-bu...