Shannonstryhn6120

4; 95% CI 1.7, 3.3), exclusive alcohol use (AOR 1.5; 95% CI 1.2, 1.8), and polysubstance use (AOR 3.9; 95% CI 2.8, 5.6 including tobacco and AOR 1.8; 95% CI 1.1, 3.0 excluding tobacco) compared with W2 never substance use. In this national study, tobacco susceptibility identified U.S. youth at risk for onset of tobacco and other substances, perhaps reflecting common etiology and clustering of substance use in youth. Identifying and preventing tobacco-susceptible youth from progressing to using addictive substances must remain a public health priority. The crystal structure of monomeric red fluorescent protein FusionRed (λex/λem 580/608 mn) has been determined at 1.09 Å resolution and revealed two alternative routes of post-translational chemistry, resulting in distinctly different products. The refinement occupancies suggest the 6040 ratio of the mature Met63-Tyr64-Gly65 chromophore and uncyclized chromophore-forming tripeptide with the protein backbone cleaved between Met63 and the preceding Phe62 and oxidized Cα-Cβ bond of Tyr64. We analyzed the structures of FusionRed and several related red fluorescent proteins, identified structural elements causing hydrolysis of the peptide bond, and verified their impact by single point mutagenesis. These findings advance the understanding of the post-translational chemistry of GFP-like fluorescent proteins beyond the canonical cyclization-dehydration-oxidation mechanism. They also show that impaired cyclization does not prevent chromophore-forming tripeptide from further transformations enabled by the same set of catalytic residues. Our mutagenesis efforts resulted in inhibition of the peptide backbone cleavage, and a FusionRed variant with ~30% improved effective brightness. Ethylene (ethene, C2H4) was introduced into V-type crystalline starches (V-starches, namely V6- and V7-types) with different single helix contents (8.35-35.54%) by a solid encapsulation method. The morphological and structural properties of V-starches and their inclusion complexes (ICs) were characterized. The V-starches prepared with n-propanol (1-propanol) and ethanol showed a V6-type crystalline structure, whereas V-starches prepared with isopropanol (2-propanol) and t-butanol (2-methyl-2-propanol) exhibited V7-type crystalline structure. The ethylene concentrations in ICs followed the order of Vn-propanol > Vethanol > Visopropanol > Vt-butanol > Vmethanol, which appeared to be associated with the diameter of the V-starch single helices. Compared with V7-type starches, ethylene was more effectively encapsulated into V6-type starches with a smaller inner diameter. The controlled-release characteristics of ICs showed a diffusion-limited mechanism and first-order kinetics for ICs in different temperatures and relative humidities, respectively. Ethylene encapsulated in V6-type-ICs showed better controlled release kinetics than V7-type-ICs, except for Vt-butanol-IC, which probably resulted from residual t-butanol in the single helices interfering with the release of ethylene. V-starches (particularly V6-type) appear to be effective gas storage and release systems for ethylene and this encapsulation technology should enable precisely-controlled and targeted applications of ethylene for food processing and agricultural applications. Since β-cyclodextrin (β-CD) was the competitive inhibitor of pullulanase, substrate utilization and product inhibition were a conflict in β-CD production when using pullulanase for debranching to release more convertible substrate. Here, a phenylalanine complexed with cyclodextrin ligand via classic hydrophobic interaction and hydrogen bond in the crystal structure of pullulanase and absolutely conserved in all homologous, was selected to study its contribution in inhibition and to get mutants with lower affinity to β-CD. Mutants were generated by substituting phenylalanine with tyrosine, valine, histidine, cysteine, aspartic acid or alanine. Enzyme activity assay and ITC data proved that F476 mutants successfully reduced the protein affinity to β-CD. Although circular dichroism spectra and fluorescence spectroscopy results showed F476 mutation affected the secondary structure and microenvironment of pullulanase, mutants F476C and F476H retained relatively higher enzyme activity and significantly decreased affinity to β-CD. The conversion rate of starch into β-CD increased by 6.0% and 6.6%, when adding F476C or F476H in the production process. Glioblastoma (GBM) is the most malignant central nervous system tumor, with poor prognosis. Temozolomide (TMZ) has been used as a first-line drug for the treatment of GBM for over a decade, but its treatment benefits are limited by acquired resistance. Polysaccharides from Cibotium barometz (CBPs) are polysaccharides purified from the root of Cibotium barometz (L.) J. Sm., possessing sensitizing activity. The purpose of this study was to investigate the anti-cancer effect of CBP from different processing methods on U87 cells using a 1H NMR-based metabolic approach, complemented with qRT-PCR and flow cytometry, to identify potential markers and discover the targets to explore the underlying mechanism. Cibotium barometz is usually processed under sand heating in clinical applications. Polysaccharides from both the processed (PCBP) and raw (RCBP) C. barometz were prepared, and the effect on enhancing the sensitivity to TMZ was investigated in vitro. CBP can significantly increase the toxicity of TMZ to the U87 cell line, promote apoptosis, enhance cell cycle changes, and arrest cells in S phase, and RCBP demonstrated better activity. Multivariate statistical analyses, such as principal component analysis (PCA) and orthogonal projection to latent structure with discriminant analysis (OPLS-DA), were used to identify metabolic biomarkers, and 12 metabolites in the cell extract samples were clearly identified as altered after RCBP exposure. NMR-based cell metabolomics provided a holistic method for the identification of CBP's apoptosis-enhancing mechanisms and the exploration of its potential applications in preclinical and clinical studies. V.The necessity to increase the efficiency of organ preservation has pushed physicians to consider the use of pharmacological additives in preservation solutions to minimize ischemia reperfusion injury. Here, we evaluated the effect of fucoidan, sulfated polysaccharide from brown seaweed, as an additive to IGL-1 (Institut Georges Lopez) preservation solution. Livers from Wistar rats were preserved for 24 h at 4 °C in IGL-1 solution, enriched or not with fucoidan (100 mg/L). Thereafter, they were subjected to reperfusion (2 h, at 37 °C) using an isolated perfused rat liver model. The addition of fucoidan to IGL-1 solution reduced hepatic injury (AST, ALT) and improved liver function compared to IGL-1 solution without fucoidan. In addition, we noted a significant increase in the phosphorylation of AMPK, AKT protein kinase and GSK3-β, leading to a reduction in VDAC phosphorylation, as well as a reduction in apoptosis (caspase 3), mitochondrial damage, oxidative stress and endoplasmic reticulum (ER) stress markers. Furthermore, ERK1/2 and P38 MAPKs phosphorylation significantly decreased after supplementation of IGL-1 solution with fucoidan. In conclusion, the supplementation of IGL-1 solution with fucoidan maintained liver graft integrity and function through the prevention of the ER stress, oxidative stress and mitochondrial dysfunction. Fucoidan could be considered as potential natural therapeutic agent to alleviate graft injury. Toll-like receptors (TLRs) play important roles in the innate system by recognizing pathogen-associated molecular patterns derived from various microbes. In this study, we reported the cloning and identification of paTLR3 and paTLR4 interactor with leucine rich repeats (TRIL) cDNA from silvery pomfret (Pampus argenteus). The full-length paTLR3 and paTRIL cDNA were 2996 and 3163 bp long, respectively. Both of the two proteins contained many LRR domains, one LRR-C terminal domain and one transmembrane region, which fits with the characteristic TLR and its analogue domain architecture. Phylogenetic analyses revealed that paTLR3 and paTRIL shared the closest relationship with Lateolabrax japonicas and Notothenia coriiceps, respectively. The expression levels of paTLR3 and paTRIL varied greatly among the examined tissues with the highest expression both in liver. Following exposure to V. anguillarum flagellin, A. hydrophila lipopolysaccharide (LPS) and L. plantarum lipoteichoic acid (LTA), paTLR3 and paTRIL were all up-regulated. V. anguillarum flagellin induced the highest expression levels of paTLR3 and paTRIL. A. hydrophila flagellin and A. hydrophila LPS induced the highest expression levels of IL-1β and IL-8, respectively. The present results will provide the valuable information for understanding the structure, function and the immune defense process of paTLR3 and paTRIL in silvery pomfret. V.AIMS Opioids (i.e. morphine) were found to induce triple negative breast cancer (TNBC) metastasis while nonsteroidal anti-inflammatory drugs (i.e. Selleck 4-PBA ketolorac) were associated with decreased metastasis in TNBC. These contradictory findings demand clarification on the effect of postoperative morphine and ketorolac on TNBC metastasis. MATERIALS AND METHODS TNBC xenograft mice were established using MDA-MB-231 cells. When tumors reached ~100 mm3, the primary tumor was resected. Mice were then randomly assigned to four groups (n = 14) (i) saline, (ii) morphine (10 mg kg-1) (iii) morphine + ketorolac (10 mg kg-1 of morphine and 20 mg kg-1 of ketorolac) (iv) ketorolac (20 mg kg-1); administrated for three consecutive days after resection. Three weeks after resection, the number of lung metastases was measured. Microvessel density, thrombospondin-1 (TSP-1) and c-Myc expression in recurrent tumors were determined. To elucidate the above phenomenon in vitro, MDA-MB-231 cells were treated according to the regiment above; with or without supplementation of an AKT inhibitor to determine the activation of PI3K/AKT/c-Myc pathway. KEY FINDINGS In mice, morphine promoted TNBC metastasis and angiogenesis, decreased TSP-1 expression and increased c-Myc expression, while co-administration of ketorolac significantly reversed the phenotypes above (p  less then  .05). Mechanistically, morphine inhibited TSP-1 secretion by activating PI3K/AKT/c-Myc pathway (p  less then  .05), while ketorolac promoted TSP-1 secretion (p  less then  .05) by suppressing PI3K/AKT/c-Myc pathway. SIGNIFICANCE Our study indicated that morphine enhanced TNBC metastasis and angiogenesis while ketorolac suppressed this effect. Mechanistically, this may be related to the enhancement of TSP-1 synthesis after ketorolac administration which further de-activated PI3K/AKT/c-Myc pathway. AIMS Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) have been associated with risk factors for metabolic syndrome (MetS). Our objective was to evaluate the effect of nicotinamide (NAM) on the activities, expression and protein content of cholinesterases in a MetS model. MAIN METHODS MetS was induced in male rats administrating 40% fructose to the drinking water for 16 weeks. Additionally, from 5th week onward, the carbohydrate solution was replaced by NAM, at several concentrations for 5 h each morning for the next 12 weeks. In the 15th week, the glucose tolerance test was conducted, and blood pressure was measured. After the treatment period had concluded, the biochemical profile; oxidant stress; proinflammatory markers; and the activity, quantity and expression of cholinesterases were evaluated, and molecular docking analysis was performed. KEY FINDINGS The MetS group showed anthropometric, hemodynamic and biochemical alterations and increased cholinesterase activity, inflammation and stress markers.