Benchmarking to the Gold Standard: Hyaluronan‐Oxime Hydrogels Recapitulate Xenograft Models with In Vitro Breast Cancer Spheroid Culture

Many 3D in vitro models induce breast cancer spheroid formation; however, this alone does not recapitulate the complex in vivo phenotype. To effectively screen therapeutics, it is urgently needed to validate in vitro cancer spheroid models against the gold standard of xenografts. A new oxime‐crosslinked hyaluronan (HA) hydrogel is designed, manipulating gelation rate and mechanical properties to grow breast cancer spheroids in 3D. This HA‐oxime breast cancer model maintains the gene expression profile most similar to that of tumor xenografts based on a pan‐cancer gene expression profile (comprising 730 genes) of three different human breast cancer subtypes compared to Matrigel or conventional 2D culture. Differences in gene expression between breast cancer cultures in HA‐oxime versus Matrigel or 2D are confirmed for 12 canonical pathways by gene set variation analysis. Importantly, drug response is dependent on the culture method. Breast cancer cells respond better to the Rac inhibitor (EHT‐1864) and the PI3K inhibitor (AZD6482) when cultured in HA‐oxime versus Matrigel. This study demonstrates the superiority of an HA‐based hydrogel as a platform for in vitro breast cancer culture of both primary, patient‐derived cells and cell lines, and provides a hydrogel culture model that closely matches that in vivo.     

Functional and health-endorsing properties of wheat and barley cell wall’s non-starch polysaccharides

Cereals have captured global importance owing to the presence of bioactive moieties in cell wall. Numerous components have been considered but non-starch polysaccharides (NSP) of cereals cell wall are of prime concern. In this comprehensive review, the basic aim is to elaborate the functional and nutritional importance of cereals cell wall with special reference to NSP. Among bioactive components of cell wall, NSP, such as arabinoxylans, ß-glucans, and arabinogalactans of wheat and barley, have gained much importance. Moreover, literature revealed that NSP have greater role as prebiotic, immunomodulator, antioxidant, anti-diabetic, and cardio-protector as well as major food applications.     

Computationally Empowered Workflow Identifies Novel Covalent Allosteric Binders for KRASG12C

Due to its frequent mutations in multiple lethal cancers, KRAS is one of the most-studied anticancer targets nowadays. Since the discovery of the druggable allosteric binding site containing a G12C mutation, KRAS^G12C has been the focus of attention in oncology research. We report here a computationally driven approach aimed at identifying novel and selective KRAS^G12C covalent inhibitors. The workflow involved initial enumeration of virtual molecules tailored for the KRAS allosteric binding site. Tools such as pharmacophore modeling, docking, and free-energy perturbations were deployed to prioritize the compounds with the best profiles. The synthesized naphthyridinone scaffold showed the ability to react with G12C and inhibit KRAS^G12C. Analogues were prepared to establish structure-activity relationships, while molecular dynamics simulations and crystallization of the inhibitor-KRAS^G12C complex highlighted an unprecedented binding mode.     

Bidirectional transepithelial water transport: measurement and governing mechanisms

In the search for the mechanisms whereby water is transported across biological membranes, we hypothesized that in the airways, the hydration of the periciliary fluid layer is regulated by luminal-to-basolateral water transport coupled to active transepithelial sodium transport. The luminal-to-basolateral (JWL-->B) and the basolateral-to-luminal (JWB-->L) transepithelial water fluxes across ovine tracheal epithelia were measured simultaneously. The JWL-->B (6.1 microliter/min/cm2) was larger than JWB-->L (4.5 microliter/min/cm2, p < 0.05, n = 30). The corresponding water diffusional permeabilities were PdL-->B = 1.0 x 10(-4) cm/s and PdB-->L = 7.5 x 10(-5) cm/s. The activation energy (Ea) of JWL-->B (11.6 kcal/mol) was larger than the Ea of JWB-->L (6.5 kcal/mol, p < 0.05, n = 5). Acetylstrophanthidin (100 microM basolateral) reduced JWL-->B from 6.1 to 4.4 microliter/min/cm2 (p < 0. 05, n = 5) and abolished the PD. Amiloride (10 microM luminal) reduced JWL-->B from 5.7 to 3.7 microliter/min/cm2 (p < 0.05, n = 5) and reduced PD by 44%. Neither of these agents significantly changed JWB-->L. These data indicate that in tracheal epithelia under homeostatic conditions, JWB-->L was dominated by diffusion (Ea = 4.6 kcal/mol), whereas approximately 30% of JWL-->B was coupled to the active Na+,K+-ATPase pump (Ea = 27 kcal/mol).     

Multiple alpha-glucoside transporter genes in brewer's yeast

Maltose and maltotriose are the two most abundant fermentable sugars in brewer's wort, and the rate of uptake of these sugars by brewer's yeast can have a major impact on fermentation performance. In spite of this, no information is currently available on the genetics of maltose and maltotriose uptake in brewing strains of yeast. In this work, we studied 30 brewing strains of yeast (5 ale strains and 25 lager strains) with the aim of examining the alleles of maltose and maltotriose transporter genes contained by them. To do this, we hybridized gene probes to chromosome blots. Studies performed with laboratory strains have shown that maltose utilization is conferred by any one of five unlinked but highly homologous MAL loci (MAL1 to MAL4 and MAL6). Gene 1 at each locus encodes a maltose transporter. All of the strains of brewer's yeast examined except two were found to contain MAL11 and MAL31 sequences, and only one of these strains lacked MAL41. MAL21 was not present in the five ale strains and 12 of the lager strains. MAL61 was not found in any of the yeast strains. In three of the lager strains, there was evidence that MAL transporter gene sequences occurred on chromosomes other than those known to carry MAL loci. Sequences corresponding to the AGT1 gene, which encodes a transporter of several alpha-glucosides, including maltose and maltotriose, were detected in all but one of the yeast strains. Homologues of AGT1 were identified in three of the lager strains, and two of these homologues were mapped, one to chromosome II and the other to chromosome XI. AGT1 appears to be a member of a family of closely related genes, which may have arisen in brewer's yeast in response to selective pressure.     

The Effect of Fibre Surface Treatment on the Failure of Continuous Carbon Fibre/Epoxy Resin Composites

The failure mechanisms of a composite, consisting of continuous, aligned, high strength, polyacrylonitrile (PAN) based carbon fibre in an epoxy resin, under uniaxial tension, have been studied. In order to study the effect of the interphase/interface strength, six different levels of an electrochemical fibre surface treatment were used. Single tows containing approximately 12,000 treated carbon fibres were impregnated to produce composite rods with a fibre volume fraction of 0.55. Lengths of this impregnated tow were also set in the centre of glass-fibre/epoxy resin composite coupons which were used to study the mechanisms of failure of the embedded tows. Acoustic emission was used to monitor all samples and bundle failure was found to occur after a build-up of sub-critical damage events as previously modelled.¹ Microdebond tests demonstrated an initial increase of interfacial strength which levelled out at the higher levels. In impregnated samples with high surface treatments, catastrophic failure occurred with the crack propagating approximately perpendicular to the fibre direction. However, in samples with lower fibre surface treatments, longitudinal splitting (not accounted for in current models), occurred, meaning that a greater length of composite was involved in the final failure process. Acoustic emission has been shown to have an approximately direct relation with the predicted number of single fibre breaks in composite test-pieces; however, there was no significant difference attributable to the different surface treatments. The hybrid test coupons allow a detailed assessment of the failure mechanisms within the impregnated carbon tow. The failure strains of the embedded tow is some 5% higher than that of unsupported tow. The Weibull modulus is of the same order.