Ain 93m

Two diets were used for this study: The control diet (CD) was a defined formulation (AIN93M, Teklad, Madison, WI) while the DP diet was composed of the AIN93M diet supplemented with 25% by weight dried plum powder (from the California Dried Plum Board) and was compounded by Teklad as described previously^{35 (link),37 (link),38 (link),43 (link),58 (link),72 (link),73 (link)}.

Male SCID Hairless Outbred mice (SHO, Crl:SHO-Prkdc^SCIDHr^hr) were purchased from Charles River (Willington, MA) and maintained on a purified irradiated maintenance diet (AIN-93 M) from Harlan Teklad (Madison, WI). All experimental procedures for animal use were previously reviewed and approved by the Institutional Animal Care and Use Committee at the University of Nebraska-Lincoln and conducted in accordance with the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health.
Six-week-old mice were implanted subcutaneously with 22Rv1 (PSMA positive, right flank) and PC3 M-LN4 (PSMA negative, left flank) cells (1 × 10⁶ cells/100 μL saline) and maintained until tumor size reached approximately 3 mm in diameter. At this time, tumor-bearing animals received either 1x PBS (100 μL), IRDye 800CW carboxylate (1 nmol), IgG 800CW (IgG 800CW, 75 μg), YC-27 800CW (1 nmol), or PSMA 800CW (75 μg) injected via the tail vein. Three doses, 0.25, 0.5, and 1.0 nmol, were evaluated in mice with 22Rv1 tumors (n = 3 per dose). Specificity was further confirmed by inhibition with 2-PMPA. For this experiment, preinjection of 2-PMPA (2 μg, intravenous administration) was followed by YC-27 800CW dose (0.5 nmole). Images were captured 24 h after injection of all compounds using the Pearl Impulse Small Animal imaging system.

C57BL/6J male mice were assigned randomly to one of two groups (n = 12 total, n = 6/group, n = 3/cage, n = 2 cages/group) and proffered either the CON or the mSAD for a period of 15 weeks. The CON (AIN-93M, TD.94048, Envigo, Madison, WI, United States) is formulated for optimal health and is a common standard reference diet used in rodent research (7 (link)). AIN-93M consists of 12.4% protein (13.7% kcal), 68.3% carbohydrate (75.9% kcal), and 4.1% fat (10.3% kcal) (Table 1). The modified Standard American diet (mSAD, TD.180061, Envigo, Madison, WI, United States) was formulated to approximate 50th percentile macronutrient intakes from data reported by NHANES. Macro- and micronutrient levels are listed in Table 1 and Table 2, respectively. The mSAD is composed of 12.3% protein dry matter (12.2% kcal), 49.5% carbohydrate (49.2% kcal), and 17.3% fat (38.6% kcal) (Table 1). The mSAD contains HFCS as the primary source of carbohydrate at a level of 26% of the diet (by weight). Multiple sources of saturated and unsaturated fat were incorporated in the mSAD, including palm oil, soybean oil, corn oil, cottonseed oil, lard, beef tallow, and anhydrous milkfat (Table 1). Moreover, the mSAD contains reduced levels of fiber and increased levels of sodium, which are supported by NHANES data (7 (link)).