Adenovirus-Mediated Inducible Expression of a PD-L1 Blocking Antibody in Combination with Macrophage Depletion Improves Survival in a Mouse Model of Peritoneal Carcinomatosis

Immune checkpoint inhibitors (ICIs) have demonstrated remarkable efficacy in a growing number of malignancies. However, overcoming primary or secondary resistances is difficult due to pharmacokinetics issues and side effects associated with high systemic exposure. Local or regional expression of monoclonal antibodies (mAbs) using gene therapy vectors can alleviate this problem. In this work, we describe a high-capacity adenoviral vector (HCA-EFZP-aPDL1) equipped with a mifepristone-inducible system for the controlled expression of an anti-programmed death ligand 1 (PD-L1) blocking antibody. The vector was tested in an immune-competent mouse model of colorectal cancer based on implantation of MC38 cells. A single local administration of HCA-EFZP-aPDL1 in subcutaneous lesions led to a significant reduction in tumor growth with minimal release of the antibody in the circulation. When the vector was tested in a more stringent setting (rapidly progressing peritoneal carcinomatosis), the antitumor effect was marginal even in combination with other immune-stimulatory agents such as polyinosinic-polycytidylic acid (pI:C), blocking mAbs for T cell immunoglobulin, mucin-domain containing-3 (TIM-3) or agonistic mAbs for 4-1BB (CD137). In contrast, macrophage depletion by clodronate liposomes enhanced the efficacy of HCA-EFZP-aPDL1. These results highlight the importance of addressing macrophage-associated immunoregulatory mechanisms to overcome resistance to ICIs in the context of colorectal cancer.     

Hydrogenation of unsaturated fatty esters during isobutane chemical ionization mass spectrometry

Hydrogenation of double bonds was observed to occur during the isobutane chemical ionization mass spectrometry (MS) of unsaturated fatty esters. Chemical ionization (CI) spectra of a series of methyl esters in the C16–C20 carbon range containing 0–4 double bonds showed a variety of ionization characteristics in the molecular ion cluster, including hydride abstraction, charge exchange, protonation and, for the unsaturated fatty acids (FA), hydrogenation of the double bond followed by protonation. The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

Effects of bile salts on rat hepatic acyl CoA:Cholesterol acyltransferase

Acyl CoA:cholesterol acyltransferase (EC2.3.1.26, ACAT), responsible for intracellular esterification of cholesterol, may play an important role in cholesterol trafficking within the cell, and thus, in maintenance of cellular cholesterol homeostasis. Bile acids are potential regulators of cholesterol trafficking in the liver. Therefore, the effect of bile salts on hepatic ACAT activity was studied in the perfused rat liver. ACAT activity was increased after liver perfusion with either taurocholate or taurochenodeoxycholate. However, addition of these bile salts at physiological concentrationsin vitro had little effect on microsomal ACAT activity. The increase in hepatic ACAT activity due to perfusion with bile salts was accompanied by reduced accumulation of very low density lipoprotein cholesterol in the perfusate, but there was no effect on 3-hydroxy-3-methylglutaryl-CoA reductase activity. Hepatic ACAT activity was decreased after bile diversion for four hours in the intact animal. This treatment had no statistically significant effect on 3-hydroxy-3-methylglutaryl-CoA reductase activity. These data suggest that bile salts induce changes in hepatic compartmentation and traffic of cholesterol within the hepatocyte accompanied by response of ACAT activity to maintain cellular cholesterol homeostasis.     

Functional properties of collagenous material from chicken feet

Collagenous material was obtained from chicken feet skins and tendons, defatted with ethylic ether and treated with 0.05 M and 0.6 M NaCl solutions. Part of the material was dried at 35 degrees C in a forced air convection oven and another was freeze-dried. The air dried material had 77.2 g/100 g of collagen and the freeze-dried material 76.7 g/100 g. Both dehydrated materials showed the same behaviour for gel formation and cold water holding capacity. The material air dried had higher emulsifying capacity than the freeze-dried one. The collagen of freeze-dried material had higher solubility in 0.5 M acetic acid and water at 70 degrees C than air dried material. Gel strength, emulsion stability and water holding capacity at 60 degrees C were higher for freeze-dried material. The results indicate the potential use of these materials as functional ingredients in meat products.     

Functional properties of a protein product from Caryodendron orinocense (Barinas nut)

The functional properties of Caryodendron orinocense protein product were investigated and compared with those of soybean (Glycina maxima). The product protein content was 24.47 g/100 g (Nx6.25). Solubility increased at both sides of the isoelectric point (pH 4.0) and with increased NaCl concentration up to 0.5M. Compared with soybean flour (50% protein), the protein product exhibited higher water and oil absorption, but lower emulsifying activity, emulsion stability, foaming capacity, and foam stability, the last one increase at higher pH. Emulsifying activity, foaming capacity, and foam stability were ionic strength dependent. C. orinocense protein product increased its emulsifying activity steadily from 0.05M to 0.75M NaCl, while it remained almost constant for soybean flour. Foaming capacity increased drastically at pH 10. The minimum time and concentration to form a gel was 20% in 4 min and 10% in 8 min for the Caryodendron protein product and soybean flour, respectively. The bulk density was 0.5056+/-0.0041 g/mL.     

Resins containing amino and carboxylic acid groups. Synthesis,characterization, and metal ion retention properties

The crosslinked poly[3‐(methacryloylamino)propyl]dimethyl(3‐sulfopropyl)ammonium hydroxide], P(MAPDSA), and poly[3‐(methacryloylamino)propyl]dimethyl(3‐sulfopropyl)ammonium hydroxide‐co‐acrylic acid], P(MAPDSA‐co‐AA), were synthesized by radical polymerization. The resins were completely insoluble in water. Due to the lower metal ion retention of P(MAPDSA), the metal ions investigated under competitive and noncompetitive conditions for Cu(II), Cd(II), Hg(II), Zn(II), Pb(II), and Cr(III) ions by batch and column equilibrium procedures were carried out only for P(MAPDA‐co‐AA), particularly for Hg(II). The resin–Hg(II) ion equilibrium was achieved before 15 min. The resin showed a maximum retention capacity value for Hg(II) at pH 2 of 1.89 meq/g. The resin showed a high selectivity to Hg(II) ions. The recovery of the resin was investigated at 25°C with different concentrations of HNO₃ and HClO₄. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 98: 525–530, 2005     

Determination of the molecular weight distribution of cellulose on calibrated gel columns

CMC samples obtained by gel filtration on a preparative scale were used to calibrate an agarose column. The relation between molecular weight and the peak elution volume for monodisperse samples were calculated as well as the relation between band broadening and the peak elution volume. In the calculations of the molecular weight distribution curves of the studied CMC samples, consideration was given to the variable band broadening by proper transformations of the experimental data, thus making it possible to obtain numerical solutions free from oscillations. The method admits a rapid determination of the molecular weight distribution of cellulose and related materials.