Pharmacologic Tumor PDL1 Depletion with Cefepime or Ceftazidime Promotes DNA Damage and Sensitivity to DNA-Damaging Agents

The interaction between tumor surface-expressed PDL1 and immune cell PD1 for the evasion of antitumor immunity is well established and is targeted by FDA-approved anti-PDL1 and anti-PD1 antibodies. Nonetheless, recent studies highlight the immunopathogenicity of tumor-intrinsic PDL1 signals that can contribute to the resistance to targeted small molecules, cytotoxic chemotherapy, and αPD1 immunotherapy. As genetic PDL1 depletion is not currently clinically tractable, we screened FDA-approved drugs to identify those that significantly deplete tumor PDL1. Among the candidates, we identified the β-lactam cephalosporin antibiotic cefepime as a tumor PDL1-depleting drug (PDD) that increases tumor DNA damage and sensitivity to DNA-damaging agents in vitro in distinct aggressive mouse and human cancer lines, including glioblastoma multiforme, ovarian cancer, bladder cancer, and melanoma. Cefepime reduced tumor PDL1 post-translationally through ubiquitination, improved DNA-damaging-agent treatment efficacy in vivo in immune-deficient and -proficient mice, activated immunogenic tumor STING signals, and phenocopied specific genetic PDL1 depletion effects. The β-lactam ring and its antibiotic properties did not appear contributory to PDL1 depletion or to these treatment effects, and the related cephalosporin ceftazidime produced similar effects. Our findings highlight the rapidly translated potential for PDDs to inhibit tumor-intrinsic PDL1 signals and improve DNA-damaging agents and immunotherapy efficacy.     

Lung Extracellular Matrix Hydrogels Enhance Preservation of Type II Phenotype in Primary Alveolar Epithelial Cells

One of the main limitations of in vitro studies on lung diseases is the difficulty of maintaining the type II phenotype of alveolar epithelial cells in culture. This fact has previously been related to the translocation of the mechanosensing Yes-associated protein (YAP) to the nuclei and Rho signaling pathway. In this work, we aimed to culture and subculture primary alveolar type II cells on extracellular matrix lung-derived hydrogels to assess their suitability for phenotype maintenance. Cells cultured on lung hydrogels formed monolayers and maintained type II phenotype for a longer time as compared with those conventionally cultured. Interestingly, cells successfully grew when they were subsequently cultured on a dish. Moreover, cells cultured on a plate showed the active form of the YAP protein and the formation of stress fibers and focal adhesions. The results of chemically inhibiting the Rho pathway strongly suggest that this is one of the mechanisms by which the hydrogel promotes type II phenotype maintenance. These results regarding protein expression strongly suggest that the chemical and biophysical properties of the hydrogel have a considerable impact on the transition from ATII to ATI phenotypes. In conclusion, culturing primary alveolar epithelial cells on lung ECM-derived hydrogels may facilitate the prolonged culturing of these cells, and thus help in the research on lung diseases.     

Solid-phase receptor-based assay for the detection of cyclic imines by chemiluminescence, fluorescence, or colorimetry

The spirolides and gymnodimines are marine phycotoxins included in the group of cyclic imines. The toxicity of these compounds to humans is still unknown, although their toxicity by intraperitoneal injection in rodents is very high. A receptor-based method was developed using the competition of the 13-desmethyl spirolide C with biotin-labeled α-bungarotoxin for binding to nicotinic acetylcholine receptors and the immobilization of the α-bungarotoxin-receptor complex on streptavidin-coated surfaces. The quantification of the immobilized receptor can be achieved using a specific antibody. Finally, after the addition of a secondary antibody labeled with horseradish peroxidase, three alternative substrates of this enzyme generate a chemiluminescent, fluorescent, or colorimetric signal. The assay performs well in shellfish extracts and the detection range is 5-150 nM of 13-desmethyl spirolide C in shellfish extracts, which is at least 5 times more sensitive than the existing fluorescence polarization assay. This assay can also detect gymnodimine, although with 10 times lower sensitivity than the spirolide. The detection of cyclic imines with microplate assays would be useful for screening purposes in order to reduce the number of samples to be processed by bioassays or analytical methods.     

Field-induced phase transition and relaxor character in submicrometer-structured lead-free (Bi0.5Na0.5)0.94Ba0.06TiO3 piezoceramics at the morphotropic phase boundary

Submicrometer-structured (Bi(0.5)Na(0.5))(0.94)Ba(0.06)TiO(3) ceramics ((G) < 720 nm) from nanopowders were studied. The real part of the optimum room temperature set of piezoelectric coefficients obtained from resonances of the BNBT6 dense ceramic disks and shear plates [d(31) = (-37 + 1.33i) pC·N(-1), d(15) = (158.3 - 8.31i) pC·N(-1), k(t) = 40.4%, k(p) = 26.8%, and k(15) = 40.2%] and d(33) (148 pC·N(-1)) can be compared with the reported properties for coarse-grained ceramics. Shear resonance of thickness-poled plates is observed at T = 140°C. Permittivity versus temperature curves of poled samples show relaxor character up to T(i) = 230°C on heating and T(i) = 210°C on cooling of the depoled samples. The phase transition from the room-temperature ferroelectric (FE) to a low-temperature non-polar at zero field (LTNPZF) phase can be observed as a sharp jump in ε(δ)(33)'(T) curves or, as the degree of poling decreases, as a soft change of slope of the curves at T(FE-LTNPZF) = T(d) = 100°C. This dielectric anomaly is not observed on cooling of depoled samples, because the FE phase is field-induced. The observed macroscopic piezoelectric activity above T(d) is a consequence of the coexistence of nanoregions of the FE phase in the interval between T(FE-LTNPZF) and T(i).     

Development of a predictive model for choosing cure agents for elastomer blends

In elastomer blends with either polarity differences or unbalanced double bond concentration, cure agents have a preferential partition to one of the phases, with a tendency of having overcure in one phase and undercure in the other. To achieve proper vulcanization and improved performance in the final product, it is essential to have a correct balance between solubility and miscibility of each of the various cure agents in the elastomer blend composite. Cure agents with varying polarities and various reactivities have different crosslinking densities in the rubber compound. A homogeneous crosslinking density leads to superior mechanical properties. The method presented in this work includes a new tool based on a genetic optimization algorithm for assessing the partitioning of cure agents in different elastomers and their blends. The quantitative data allowed for a series of analyses of the solubility of the cure agents in the elastomer mixtures of different phases and was validated by correlation with their physico–chemical and mechanical properties of the resulting blend. This is an important tool for planning a cure system in rubber blends and to avoid incorrect partitioning of cure agents and consequent uneven curing of the final compound. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41929.     

Determination of cosmetic ingredients causing extrusion‐coated and adhesive joint multilayer packaging delamination

In order to study the effect of several compounds on packaging stability, different cosmetic ingredients at two concentration levels were added to a NeoPCL® (Acofarma, Terrassa, Spain) water emulsion, and the preparations packed in sachets and stored at 40°C during 3 months. After that, the packaging was subjected to a T‐peel test and headspace solid‐phase microextraction gas chromatography mass spectrometry (HS‐SPME‐GC‐MS) analysis. The HS‐SPME‐GC‐MS analyses were performed using a 75 μm carboxen polydimethylsiloxane fibre to check for the presence of the studied analytes in the inner layers. The study revealed that the presence of a phenyl and a hydroxyl group in the compound structure lead to an important loss of adhesion between packaging layers joined by an adhesive. The interaction between the cosmetic ingredient and the adhesive was proposed as the main cause of the loss of adhesion. However, extrusion‐coating packaging was more susceptible to delamination, particularly with the volatile compounds. Copyright © 2009 John Wiley & Sons, Ltd.     

Pressure drop correlation for oil–refrigerant R134a mixture flashing flow in a small diameter tube

This paper presents an experimental investigation of the ester oil ISO VG10/refrigerant R134a mixture flashing flow in a 6.0 m long, 3.22 mm ID tube, which is one of the primary steps towards the construction of a methodology for the study of the lubrication and gas leakage in refrigeration compressors. The phase change starts with solubility reduction of the refrigerant in the oil as the pressure decreases due to the friction forces. In this flashing flow the foam pattern is observed at the end of the tube as vapor quality reaches high values, and this is a particular phenomenon of this kind of mixture flow. In order to study this pressure drop, an experimental apparatus was designed to allow the measurement of both pressure and temperature profiles along the tube as well as the visualization of the flow patterns. Pressure and temperature distribution along the flow were measured for saturation pressure ranging from 450 to 650 kPa, mass flux ranging from about 2000 to 3000 kg/(m²s), temperatures around 303 K, and inlet refrigerant concentration varying between 0.2 and 0.4 kg ref/kg mixt. An available correlation proposed to predict the frictional pressure drop for a mixture composed by the mineral oil SUNISO 1GS and refrigerant R12 flowing in small diameter tubes yielded large deviations in predicting the ester oil and refrigerant R134a mixture flow. A new correlation has been proposed that fitted the experimental data with rms deviations of 24%.