Solu medrol

Methylprednisolone (Solu-Medrol: Pfizer, Sollentuna, Sweden) was administered at baseline to some animals at a 30 mg × kg⁻¹ dose to induce immunosuppression and facilitate fungal growth.

In the whole-liver perfusion model, the HA, PV, and CBD were dissected free and cannulated on the back table. Briefly, the gallbladder was removed and the cystic duct and artery was ligated. The HA was cannulated with a 12F cannula, and the PV with a 24F cannula (Organ Assist, Groningen, The Netherlands). The CBD was cannulated with a 4 mm acorn tip cannula (Medtronic, Minneapolis, MN, USA). The hepatic veins were left uncannulated. The liver was weighed immediately before and after NMP and functional data was normalized to preperfusion weight. Finally, a pre-pump flush consisting of lactated ringers (LR), 0.05 mg/ml of solumedrol (Pfizer, NYC, NY, USA), and 1 ug/L of Epoprostenol (GlaxoSmithKline, Research Triangle Park, NC, USA) was flushed with 2L through the PV and 1L through the HA. Three whole livers with tPA (labeled W1-W3) were compared to seven whole livers with no-tPA (labeled C1-C7).

GO severity and clinical activity stage were evaluated at presentation by trained investigators. GO severity was evaluated according to the EuGoGo guidelines (21 (link)). The lid fissure width was evaluated in millimeters by a router, and proptosis was evaluated with a Hertel exophthalmometer. Diplopia was classified according to the Gorman score. A complete ophthalmological evaluation was carried out by an expert ophthalmologist. The GO was defined as moderate to severe when eye disease had a sufficient impact on daily life with one or more of the following: lid retraction 2 mm or more, moderate or severe soft tissue involvement, exophthalmos three or more mm above 21, and inconstant or constant diplopia. GO was considered active when the clinical activity score (CAS) was ≥3 (22 (link)). Corticosteroid treatment for GO was administered to patients with moderate to severe active GO (21 (link)) and consisted of intravenous MPDS (Solumedrol; Pfizer, Karlsruhe, Germany) injections with a cumulative dose of 4.5 g subdivided in 12 weekly infusions (500 mg each week for 6 weeks followed by 250 mg week for 6 weeks); this treatment was initiated in all patients within 8 months after starting ATD and following the EuGoGo protocol (23 (link), 24 (link)).

Induction of profound persistent neutropenia and immunosuppression was initiated 6 days before intracisternal inoculation and maintained using cytarabine (Ara-C) (Cytarabine injection; Hospira, Inc., Lake Forest, IL) at a dosage of 440 mg/m² for 5 consecutive days (days 1-5) with subsequent administration on days 8-9 and 13-14, as previously described.²⁰ Methylprednisolone (Solu-Medrol^®, Pfizer Inc., NY, NY) was administered at 5 mg/kg of body weight for immunosuppression from day 1 to the end of experiment. Ceftazidime (75 mg/kg i.v. twice daily; Glaxo Pharmaceuticals, Division of Glaxo Inc., Research Triangle Park, NC), gentamicin (5 mg/kg i.v. every other day; Elkins-Sinn, Inc., Cherry Hill, NJ), and vancomycin (15 mg/kg i.v. daily; Abbott Laboratories, North Chicago, IL) were administered intravenously beginning on day 4 of the study to prevent bacterial infections. Vancomycin (50 mg/L) was added to the drinking water to prevent antibiotic-associated diarrhea due to Clostridium spiroforme from the beginning of induction of neutropenia.

Animals were given intraperitoneal CTLA4-Ig (Abatacept, Bristol Myers-Squibb, New York, NY, USA) or isotype Ig control (VEGF-Ig; Avastin, Genentech, South San Francisco, CA, USA) twice a week, with an initial dose of 1 mg and subsequent doses of 0.5 mg. Animals given steroids were given a single 200 µg bolus dose of methylprednisone (Solu-Medrol, Pfizer, New York, NY, USA) on day 6 posttransplantation.