Nonhypothesis Analysis of a Mutagenic Soybean (Glycine max [L.]) Population for Protein and Fatty‐Acid Composition

Soybean is a major source of oil for food, feed, and biofuel production. Mutagenesis is a tool for creating unique traits useful in breeding programs. The aim of this study is to use nonhypothesis statistical testing methods to make decisions about a mutagenic population. To this end, a total of 1037 mutation lines and 28 wild‐type lines were analyzed for fatty‐acid composition and protein content. Principal component analysis (PCA) was used to analyze the fatty acid profile, multivariate analysis of variance (MANOVA) to build a selection model for seed weight per plant and weight per 10 seeds, and clustering in conjunction with power analysis to determine the minimum number of individuals needed to create a MANOVA selection model for the oil to protein content. Five of the 35 possible entries were identified by PCA analysis for stearic acid and four of 16 possible entries for oleic acid. Interestingly, most of the selected mutants were validated genetically. In fact, selected mutants with high seed stearic acid or high seed oleic acid contents were verified to carry mutations on GmFAD2‐1A, GmFAD2‐1B, and GmSACPD‐C genes. This shows a promising method of identifying smaller portion of the population to screen for desired mutations.     

Study of Some Experimental Parameters in the Synthesis of Triacylglycerols with CLA Isomers and Structural Analysis

The present research deals with the synthesis of structured triacylglycerols (TAG) by enzymatic treatment of sn-1,3-diacylglycerol (sn-1,3-DAG) with conjugated linoleic acid (CLA) isomers using the immobilized lipase from Rhizomucor miehei (Lipozyme^® IM) under different experimental conditions. In particular, the influence of reaction parameters, such as temperature, enzymatic load, reaction time and DAG/CLA ratio has been evaluated using an experimental design software with a screening objective. Two responses have been selected, they are the percentage of CLA isomers in total TAG and in the sn-2- position and a three-level-4-factor fractional factorial experimental design was used to screen the variables. The results showed that the selected experimental variables have an influence on the enzymatic reaction, in particular, the DAG/CLA substrate ratio and the temperature, both of which inversely correlated with CLA incorporation, but also the enzymatic load and the reaction time, both directly correlated with CLA incorporation. The best results for CLA isomer % content both in total TAG (46.3%) and in the sn-2- position (52.2%) were obtained at 40 °C for 96 h, with 20% enzymatic load and a 0.5 reactive ratio.     

Commercial bentonites as detoxifier of broiler feed contaminated with aflatoxin

Some bentonites have been probed to be efficient as sequestering agents for aflatoxins; they decreased the bioavailability of the toxin in the gastrointestinal tracts of birds when they are incorporated in the diets. The binding capacity of these adsorbents varied with the rheological source and even among batches of a given source. Three bentonites from different sources in Argentina, which have very different aflatoxins (AFs) adsorption capacity, were studied. The characterization comprises chemical analysis, scanning electron microscopy (SEM), X-ray power diffraction (XRD), Hg intrusion porosimetry, swelling capacity, etc. The main factors affecting the adsorption of AFs seem to be related to the isomorphic substitution of the montmorillonite and to electrostatic interactions generated by the surface charge of the samples. Neither the mean pore size nor the percentage of quartz has any effect upon the AFs adsorption.     

Influence of Laser Glazing on the Characterization of Plasma-Sprayed YSZ Coatings

In this study, 8 wt.% yttria-stabilized zirconia powder was deposited on the substrates by atmospheric plasma spray. The coatings were post-treated by laser glazing under different parameters in order to densify them. The characterization of the laser molten pools under different laser treatment conditions was studied. Preheating processes were also employed. Scanning electron microscopy observations of the surface and cross section of as-sprayed and laser-glazed coatings were carried out to investigate the influence of laser glazing on the microstructure on laser-glazed coatings. The results show that preheating processes improve the coating in terms of deepening the laser-glazed layer, reducing the number of vertical cracks and surface density of cracks and widening the molten pool. Finally, the influences of linear energy density on the characterization of the glazed layer are discussed.     

Producing High-Quality Titanium Alloy by a Cost-Effective Route Combining Fast Heating and Hot Processing

Powder metallurgy is a very attractive method for producing titanium alloys, which can be near-net-shape formed and have freedom in composition selection. However, applications are still limited due to product affordability. In this paper, we will discuss a possible cost-effective route, combining fast heating and hot processing, to produce titanium alloys with similar or even better mechanical properties than that of ingot metallurgy titanium alloys. Two titanium alloys, Ti-5Al-5V-5Mo-3Cr (Ti-5553) and Ti-5Fe, were successfully produced from HDH titanium powder and other master alloy powders using the proposed processing route. The effect of the processing route on microstructural variation and mechanical properties have been discussed.     

The GAUGAA Motif Is Responsible for the Binding between circSMARCA5 and SRSF1 and Related Downstream Effects on Glioblastoma Multiforme Cell Migration and Angiogenic Potential

Circular RNAs (circRNAs) are a large class of RNAs with regulatory functions within cells. We recently showed that circSMARCA5 is a tumor suppressor in glioblastoma multiforme (GBM) and acts as a decoy for Serine and Arginine Rich Splicing Factor 1 (SRSF1) through six predicted binding sites (BSs). Here we characterized RNA motifs functionally involved in the interaction between circSMARCA5 and SRSF1. Three different circSMARCA5 molecules (Mut1, Mut2, Mut3), each mutated in two predicted SRSF1 BSs at once, were obtained through PCR-based replacement of wild-type (WT) BS sequences and cloned in three independent pcDNA3 vectors. Mut1 significantly decreased its capability to interact with SRSF1 as compared to WT, based on the RNA immunoprecipitation assay. In silico analysis through the “Find Individual Motif Occurrences” (FIMO) algorithm showed GAUGAA as an experimentally validated SRSF1 binding motif significantly overrepresented within both predicted SRSF1 BSs mutated in Mut1 (q-value = 0.0011). U87MG and CAS-1, transfected with Mut1, significantly increased their migration with respect to controls transfected with WT, as revealed by the cell exclusion zone assay. Immortalized human brain microvascular endothelial cells (IM-HBMEC) exposed to conditioned medium (CM) harvested from U87MG and CAS-1 transfected with Mut1 significantly sprouted more than those treated with CM harvested from U87MG and CAS-1 transfected with WT, as shown by the tube formation assay. qRT-PCR showed that the intracellular pro- to anti-angiogenic Vascular Endothelial Growth Factor A (VEGFA) mRNA isoform ratio and the amount of total VEGFA mRNA secreted in CM significantly increased in Mut1-transfected CAS-1 as compared to controls transfected with WT. Our data suggest that GAUGAA is the RNA motif responsible for the interaction between circSMARCA5 and SRSF1 as well as for the circSMARCA5-mediated control of GBM cell migration and angiogenic potential.     

1,2,4-Oxadiazole Topsentin Analogs with Antiproliferative Activity against Pancreatic Cancer Cells,Targeting GSK3β Kinase

A new series of topsentin analogs, in which the central imidazole ring of the natural lead was replaced by a 1,2,4-oxadiazole moiety, was efficiently synthesized. All derivatives were pre-screened for antiproliferative activity against the National Cancer Institute (NCI-60) cell lines panel. The five most potent compounds were further investigated in various pancreatic ductal adenocarcinoma (PDAC) cell lines, including SUIT-2, Capan-1, and Panc-1 cells, eliciting EC₅₀ values in the micromolar and sub-micromolar range, associated with significant reduction of cell migration. These remarkable results might be explained by the effects of these new topsentin analogues on epithelial-to-mesenchymal transition markers, including SNAIL-1/2 and metalloproteinase-9. Moreover, flow cytometric analysis after Annexin V-FITC and propidium iodide staining demonstrated that these derivatives enhanced apoptosis of PDAC cells. Keeping with these data, the PathScan intracellular signaling and ELISA array revealed cleavage of caspase-3 and PARP and a significant inhibition of GSK3β phosphorylation, suggesting this kinase as a potential downstream target of our novel compounds. This was further supported by a specific assay for the evaluation of GSK3β activity, showing IC₅₀ values for the most active compounds against this enzyme in the micromolar range.