Kenalog

The surgical techniques used to treat these patients have been previously described [31 (link), 34 (link)]. Conventional 25-gauge three-port pars plana vitrectomy (PPV) techniques were used. During the procedure, a diluted suspension of triamcinolone acetonide (Kenalog 40 mg/mL; Bristol-Myers Squibb, New York, NY, USA) was used to improve visualization of the posterior hyaloidal condensation (PHC) and its base. Active suction was used to pull the PHC from the superficial retina via a soft-tipped microcannula before the use of perfluorocarbon heavy liquid injection to hydropneumatically reattach the retina and drain the subretinal fluid (SRF), followed by argon laser endophotocoagulation after complete retinal reattachment. A low-lying SB was placed in a subset of cases to support the retinal edge and release the residual traction. These cases were selected based on the following findings: inferior location of the GRT, evidence of PVR grade C or worse, GRT cases with a circumferential tear extension of less than 180º, and the presence of other risk factors such as trauma, young age, high myopia, and hereditary conditions such as Marfan syndrome and Stickler syndrome [35 (link), 36 (link)]. Finally, a 15% octafluoropropane (C₃F₈) gas mixture or 5000 cs silicone oil were added as a long-lasting tamponade.

CD1 female mice, 6–8 weeks old were used in the corticosteroid (triamcinolone acetate, Kenalog) experiments. Mice were obtained from Charles River Laboratories (Raleigh, NC). For the corticosteroid model, mice were immunosuppressed with a single dose of Kenalog (Bristol-Myers Squibb Company, Princeton, NJ, USA) injected subcutaneously (s.c.) at 40 mg/kg 1 day prior to inoculation. For the immunocompetent experiments, mice 10–12 weeks old with targeted myeloid deletions of HIF1α created via crosses into a background of lysozyme M–driven cre (HIF^C) expression and littermate controls (cre-/HIF1α floxed) were used as described in [22] (link). For infections, mice were lightly anesthetized and immobilized in an upright position using rubber bands attached to a Plexiglas stand for oropharyngeal aspiration. A blunt 20G needle attached to a 1 ml syringe was advanced into the trachea to deliver the indicated number of conidia (3–7×10⁷) in a volume of 0.05 ml PBS or PBS with 0.025% Tween-20.

Standardized steroid and chemotherapeutic mouse models of invasive pulmonary aspergillosis were used for this study^{31 (link)}. For RNA-Seq experiments, female mice (CD1), 6–8 weeks of age, were housed 4 animals per cage in a controlled environment in the Dartmouth CCMR facility consisting of HEPA filtered air, autoclaved food ad libitum. Steroid model mice received a single dose of Kenalog (Bristol-Myers Squibb Company, Princeton, NJ, USA) injected subcutaneously (s.c.) at 40 mg/kg 1 day prior to inoculation. Chemotherapy model mice received intraperitoneal (i.p.) injections of cyclophosphamide (Baxter Healthcare Corporation, Deerfield, IL, USA) at 175 mg/kg 2 days prior to inoculation and a subcutaneously (s.c.) injection of Kenalog at 40 mg/kg 1 day prior to inoculation. Mice were inoculated via the intranasal route with 2 × 10⁶ 
A. fumigatus strain CEA10 (also called CBS144.89) conidia per mouse on day 0. Mock samples were inoculated with sterile phosphate buffer saline. Mouse lungs were harvested from each animal on day 2 and 3 post inoculation. All animal studies were carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institute of Health, and were approved by the respective Virginia Tech and Dartmouth IACUCs.