NOGO-A induction and localization during chick brain development indicate a role disparate from neurite outgrowth inhibition

BACKGROUND: Nogo-A, a myelin-associated protein, inhibits neurite outgrowth and abates regeneration in the adult vertebrate central nervous system (CNS) and may play a role in maintaining neural pathways once established. However, the presence of Nogo-A during early CNS development is counterintuitive and hints at an additional role for Nogo-A beyond neurite inhibition. RESULTS: We isolated chicken NOGO-A and determined its sequence. A multiple alignment of the amino acid sequence across divergent species, identified five previously undescribed, Nogo-A specific conserved regions that may be relevant for development. NOGO gene transcripts (NOGO-A, NOGO-B and NOGO-C) were differentially expressed in the CNS during development and a second NOGO-A splice variant was identified. We further localized NOGO-A expression during key phases of CNS development by in situ hybridization. CNS-associated NOGO-A was induced coincident with neural plate formation and up-regulated by FGF in the transformation of non-neural ectoderm into neural precursors. NOGO-A expression was diffuse in the neuroectoderm during the early proliferative phase of development, and migration, but localized to large projection neurons of the optic tectum and tectal-associated nuclei during architectural differentiation, lamination and network establishment. CONCLUSION: These data suggest Nogo-A plays a functional role in the determination of neural identity and/or differentiation and also appears to play a later role in the networking of large projection neurons during neurite formation and synaptogenesis. These data indicate that Nogo-A is a multifunctional protein with additional roles during CNS development that are disparate from its later role of neurite outgrowth inhibition in the adult CNS

Integrated genomic characterization of pancreatic ductal adenocarcinoma

We performed integrated genomic, transcriptomic, and proteomic profiling of 150 pancreatic ductal adenocarcinoma (PDAC) specimens, including samples with characteristic low neoplastic cellularity. Deep whole-exome sequencing revealed recurrent somatic mutations in KRAS, TP53, CDKN2A, SMAD4, RNF43, ARID1A, TGFβR2, GNAS, RREB1, and PBRM1. KRAS wild-type tumors harbored alterations in other oncogenic drivers, including GNAS, BRAF, CTNNB1, and additional RAS pathway genes. A subset of tumors harbored multiple KRAS mutations, with some showing evidence of biallelic mutations. Protein profiling identified a favorable prognosis subset with low epithelial-mesenchymal transition and high MTOR pathway scores. Associations of non-coding RNAs with tumor-specific mRNA subtypes were also identified. Our integrated multi-platform analysis reveals a complex molecular landscape of PDAC and provides a roadmap for precision medicine

Increasing Stringiness of Low Fat Mozzarella Cheese Using Polysaccharides

We examined the ability of polysaccharides to function as fat mimetics in low-fat (LF) mozzarella string cheese to improve functionality by acting like fat globules to separate protein fibers during cheese extrusion. Low-fat mozzarella cheese curd made from 273 kg of 0.7% fat milk was salted at a rate of 10 g/kg then divided into 3.6-kg batches that were hand-stretched in 5% brine at 80° C and formed into a homogeneous mass. The hot cheese was hand mixed with a hot 80° polysaccharide slurry, placed into a small piston-driven extruder and then forced through a 16-mm die to form the string cheese. Extruded string cheese was cut manually into approximately 15-cm lengths. From preliminary trials using starches (waxy corn, waxy rice, and instant tapioca starch), xanthan and guar gums, and polydextrose, we determined that LF string cheese made using xanthan gum most closely resembled commercial string cheese. LF cheese was then made using a 10% xanthan gum slurry added at 0.25%, 0.5%, 1.0%, 1.5%, and 2.0% (wt/wt) levels. Cheeses were analyzed for fat, salt, pH, and moisture. After 2 wk of 4° C storage, the cheese was analyzed for extent of stringiness by pulling apart the cheese longitudinally, visually observing and photographing the size, length and appearance of individual strings of cheese. Hardness was determined using a Warner-Bratzler shear test. A consumer liking test was conducted after 2 and 8 wk storage time at 6° C. At 2-wk storage, using a hedonic scale (1 to 9) for overall liking, the LF string cheese with 1% added xanthan slurry (score = 6.8) was liked more (PWhen considered on a JAR scale, most of the panelists scored the LF cheese with added xanthan gum (1%) as having the right texture, while only some did so for the retail cheese. The LF control cheese with no added gum was considered too firm. Using a visual comparison, adding the xanthan gum slurry produced greater fiber formation with the longest and most complete string separation. After 8 wk storage, the LF cheeses had softened extensively with fracture stress for LF cheese decreasing from 12 to 20 kg at 2 wk to 1.5 to 3 kg at 8 wk. Extent of stringiness also decreased during storage