Molecular cloning and functional characterization of the flavonoid 3′-hydroxylase gene from Rubus coreanus Miquel

Rubus coreanus Miquel is a Korean black raspberry used in folk medicine and functional foods. To investigate the biosynthesis pathway of anthocyanin in R. coreanus Miquel, the complete coding sequence of flavonoid 3′-hydroxylase (F3′H), designated as RcMF3′H1, was cloned for the first time using the Korean black raspberry transcriptome library. The deduced amino acid sequence of RcMF3′H1 contained the proline-rich “hinge” region, P450 consensus heme-binding domain, and F3′H-specific motifs. Phylogenetic analysis revealed that RcMF3′H1 was clustered into the same subgroup as other plant F3′Hs. In addition, expression analysis by quantitative real-time PCR revealed the involvement of RcMF3′H1 in methyl jasmonate-mediated anthocyanin biosynthesis. Furthermore, the ability of the RcMF3′H1 gene to complement the Arabidopsis transparent testa 7-1 mutant suggested that RcMF3′H1 encodes the functional F3′H enzyme involved in anthocyanin biosynthesis. Taken together, the cloning and molecular characterization of RcMF3′H1 will facilitate a better insight into the anthocyanin biosynthesis pathway in R. coreanus Miquel

Influence of ripening stages on phytochemical composition and bioavailability of ginseng berry (Panax ginseng C.A. Meyer)

The presence of large amounts of bioactive compounds such as saponins and flavonoids in ginseng (Panax ginseng) berry suggests its potential as a functional resource for the food and medical industries, despite the fact that been considered a useless by-products of P. ginseng. In this study, we examined the variations in the antioxidant and anti-melanogenic potential of ginseng berry during the ripening process. We found that fully ripe berry extracts (Go-S3) contained the highest level of antioxidant and anti-melanogenic activities. Phytochemical screening suggested that alterations in polyphenol contents correlated with the variation in bioactive principles of ginseng berry during the ripening process. Furthermore, results obtained by quantitative real-time PCR, western blot, tyrosinase inhibition assay and molecular docking analysis suggested that Go-S3 probably inhibits tyrosinase activity by interacting with copper-coordinating histidines and second shell residues of tyrosinase, resulting in the reduction of melanin production in α-MSH-stimulated B16F10 cells. Taken together, these finding suggest the potential of ginseng berry as a resource for functional applications in the cosmetic industries and demonstrate that fruit ripening stages have profound effects on the pharmaceutical value of ginseng berry

Identification and expression profiling of flax (Linum usitatissimum L.) polyamine oxidase genes in response to stimuli

Polyamine oxidases (PAOs) are known to be involved in either the terminal catabolism or the back conversion of polyamines, which affect a range of physiological processes, including growth, development, and stress responses. In this study, based on genome-wide analysis, we identified five putative PAO genes (LuPAO1 to LuPAO5) in flax (Linum usitatissimum L.) that contain the amino-oxidase domain and FAD-binding-domain. The expression analysis using quantitative real-time PCR revealed spatial variations in the expression of LuPAOs in different organs. In addition, the expression level of LuPAOs in the flax cell suspension culture was increased by treatment with methyl- jasmonate (MeJA) or pectin, but not with salicylic acid or chitosan. This indicates that LuPAOs might be involved in the MeJA-mediated biological activities. Taken together, our genome-wide analysis of PAO genes and expression profiling of these genes provide the first step toward the functional dissection of LuPAOs

Second Primary Glioblastoma Multiforme Following Autologous Hematopoietic Stem Cell Transplantation in a Patient with Acute Myelogenous Leukemia

Glioblastoma multiforme (GM) is one of the most aggressive primary brain tumors, and has a poor prognosis despite intensive treatment. GM is also the most malignant astrocytoma, with histopathological features that include cellular polymorphism, rapid mitotic activity, microvascular proliferation, and necrosis. The causes of GM remain obscure, but several reports have shown associations between GM and genetic alterations and radiation exposure. Furthermore, high-dose chemotherapy/radiotherapy with autologous stem cell transplantation is increasingly being used to treat patients with leukemia, and patients who undergo stem cell transplantation have a higher risk of solid tumor cancer development later in life. Based on these associations, we discuss GM development in a patient who underwent chemoradiotherapy conditioning prior to stem cell transplantation

Serial Measurement of WT1 Expression and Decrement Ratio Until Hematopoietic Cell Transplantation as a Marker of Residual Disease in Patients with Cytogenetically Normal Acute Myelogenous Leukemia

AbstractUsing real-time quantitative PCR, we monitored Wilms tumor gene 1 (WT1) expression from diagnosis to hematopoietic stem cell transplantation (HSCT) in adult patients with cytogenetically normal acute myelogenous leukemia (CN-AML) and FLT3-ITD and NPM1 mutations. The values at diagnosis were evaluated in 104 patients. Data collected after induction chemotherapy were available for all patients, but only 68 patients were treated with HSCT. Significant WT1 expression cut-offs were determined by receiver operation characteristic curve analysis, and rates of overall survival (OS) and disease-free survival (DFS) were estimated. WT1 decrement ratios (DR) at postinduction chemotherapy and at pre- and post-HSCT compared with the diagnostic level were calculated. Higher WT1 expression at diagnosis, postinduction chemotherapy, and pre-HSCT showed inferior OS (P = .015, <.001, and .002) and DFS (P = .006, <.001, and .003). The cut-offs were determined at the median for diagnostic WT1 expression and at the 25% level from the top for other time points excluding post-HSCT. The WT1 DR ≥ 1-log after induction chemotherapy showed superior OS and DFS (P = .009 and .002) and WT1 DR ≥ 1-log preceding HSCT also showed superior OS and DFS (P = .009 and .003). Results of WT1 DR were consistently applicable in each subgroup with higher (≥1.0) and lower (<1.0) WT1 expression at diagnosis and also in NPM1-wild-type/FLT3-ITD–negative CN-AML. The WT1 DR therefore predicted survival outcomes after HSCT more accurately than did the diagnostic WT1 expression. WT1 expression may serve as a reliable marker for residual disease and WT1 DR as a prognostic indicator, particularly in NPM1-wild-type/FLT3-ITD–negative CN-AML. These measures may be applied throughout the course of treatment and even after HSCT

Transtorming Growth Factor β1 Induces Epithelial-to-Mesenchymal Transition of A549 Cells

Idiopathic pulmonary fibrosis (IPF) comprises an aggregate of mesenchymal cells. However, the cellular origin of these mesenchymal phenotypes remains unclear. Transforming growth factor β1 (TGF-β1) has been known as the main cytokine involved in the pathogenesis of IPF. We examined whether the potent fibrogenic cytokine TGF-β1 could induce the epithelial-to-mesenchymal transition (EMT) in the human alveolar epithelial cell line, A549, and determined whether snail expression is associated with the phenotypic changes observed in the A549 cells. EMT was investigated with cells morphology changes under phase-contrast microscopy, western blotting, and indirect immunofluorescence stains. E-cadherin and transcription factor, snail, were also evaluated by measuring mRNA levels using reverse transcriptase-polymerase chain rection (RT-PCR) analysis. The data showed that TGF-β1 induced A549 cells with epithelial cell characteristics to undergo EMT in a concentration-dependent manner. Following TGF-β1 treatment, A549 cells induced EMT characterized by cells morphological changes, loss of epithelial markers E-caherin and cytokeratin, increased stress fiber reorganization by F-actin, and cytokeratin replacement by vimentin. Although IL-1β failed to induce A549 cells to undergo EMT, the combination of TGF-β1 and IL-1β showed synergy effects in cells morphology changes and the expression of mesenchymal markers. The snail expression study using RT-PCR analysis provided that loss of E-cadherin expression was associated with snail expression. Stimulation of A54 cells with TGF-β1 plus IL-1β revealed a higher level of snail expression. Our data showed that EMT of A549 cells might be closely associated with snail expression

<i>De novo</i> transcriptomic analysis to reveal functional genes involved in triterpenoid saponin biosynthesis in <i>Oplopanax elatus</i> NAKAI

Oplopanax elatus is a valuable medicinal plant in the family Araliaceae that recommends itself as source of herbal preparations. Although, as in ginseng, triterpenoid saponins make up the major bioactive component of O. elatus, nothing is known about the genes that are involved in the biosynthesis of these complex compounds, as reflected also by a lack of genomic information in public databases. Using Illumina paired-end sequencing technology, we have therefore generated a transcriptome library of O. elatus from a pooled RNA sample from different organs. 208,959 unigenes were assembled from approximately 77 million high-quality reads, and 110,202 unigenes (52.7% of the unigenes) were annotated. In addition, 47,273 cDNA-derived SSRs in 38,446 unigenes were identified as potential molecular markers. Furthermore, 122 unigenes encoding 47 putative enzymes related to the biosynthesis of the backbone of triterpenoid saponins were identified by analyzing our library. The organ-specific expression of selected genes suggests that the leaves of O. elatus are the main site of triterpenoid saponin biosynthesis. The transcriptome data reported here provides valuable and comprehensive information for further research into the metabolic pathways of O. elatus as well as into genetic variation in the Oplopanax genus.   The online version of this article (doi: 10.5073/JABFQ.2017.090.004) contains supplementary files