Search

The poor prognosis for pancreatic ductal adenocarcinoma (PDAC) patients is due in part to the highly fibrotic nature of the tumors that impedes delivery of therapeutics, including nanoparticles (NPs). Our prior studies demonstrated that proglumide, a cholecystokinin receptor (CCKR) antagonist, reduced fibrosis pervading PanIN lesions in mice. Here, we further detail how the reduced fibrosis elicited by proglumide achieves the normalization of the desmoplastic tumor microenvironment (TME) and improves nanoparticle uptake. One week following the orthotopic injection of PDAC cells, mice were randomized to normal or proglumide-treated water for 3-6 weeks. Tumors were analyzed ex vivo for fibrosis, vascularity, stellate cell activation, vascular patency, and nanoparticle distribution. The histological staining and three-dimensional imaging of tumors each indicated a reduction in stromal collagen in proglumide-treated mice. Proglumide treatment increased tumor vascularity and decreased the activation of cancer-associated fibroblasts (CAFs). Additionally, PANC-1 cells with the shRNA-mediated knockdown of the CCK2 receptor showed an even greater reduction in collagen, indicating the CCK2 receptors on tumor cells contribute to the desmoplastic TME. Proglumide-mediated reduction in fibrosis also led to functional changes in the TME as evidenced by the enhanced intra-tumoral distribution of small (<12 nm) Rhodamine-loaded nanoparticles. The documented in vivo, tumor cell-intrinsic anti-fibrotic effects of CCK2R blockade in both an immunocompetent syngeneic murine PDAC model as well as a human PDAC xenograft model demonstrates that CCK2R antagonists, such as proglumide, can improve the delivery of nano-encapsulated therapeutics or imaging agents to pancreatic tumors.

The utilization and conversion of glucose to volatile acids were monitored in anaerobic digestors by C-nuclear magnetic resonance. Glucose was converted to lactate and acetate. Lactate was subsequently converted to propionate. The distribution of the labeled carbons in propionate suggested that minor amounts were produced via the randomizing pathway and that the major portion of propionate was derived from lactate.

Methane production from added substrate per se was approximately twofold greater from delignified straw than that from untreated straw when fermented with cattle manure.

Anaerobic digestors were fed daily with dairy cattle manure (5% total solids) augmented with 0-20 mM glucose and were monitored daily for gas volume and composition and volatile acid content. Propionate accumulated in digestors that were fed glucose at initial digestor concentrations of 10 mM or more. Digestors that received 14-20 mM glucose failed, but identical digestors that received 20 mM glucose plus 10 mM acetate or HCO-3 did not fail. The sparing effect of HCO-3 was primarily buffering and the similar behavior of digestors that received acetate suggest that acetate metabolism perhaps provided additional HCO-3 for buffering. Analyses of H2 and volatile acid concentrations during a 6-8-h period following feeding in digestors fed glucose or glucose plus acetate showed that propionate and H2 accumulated simultaneously and that H2 concentrations were 3 microM or less. Monitoring 13C-labeled glucose metabolism via 13C nuclear magnetic resonance indicated that glucose was primarily converted to lactate and that the major product from lactate was propionate in both glucose and glucose plus acetate fermentations.