Metabolic characterization of green pods from Vanilla planifolia accessions grown in La Réunion

Large phenotypic variation has been observed between the cultivated vanillas since a single genetic source of Vanilla planifolia was spread to the Indian Ocean and the Indonesia in the 19th century. In order to differentiate the cultivated vanilla plants, genetic studies have been conducted in the past on the plants grown in various regions such as the French island, La Réunion. However, the genetic difference was not big enough to differentiate diverse accessions of V. planifolia. In this study, metabolomics, in which genetic variation could be amplified, was employed to delve into the variation between the cultivated vanilla plants. To obtain a broad view of the metabolome, nuclear magnetic resonance (NMR) spectroscopy was applied to the analysis of V. planifolia green pods. Principal component analysis (PCA) and partial least square-discriminant analysis (PLS-DA) of the data showed that the accessions could be differentiated according to their glucovanillin and glucosides A and B contents. Furthermore, a correlation between the glucovanillin content and the pod length, number of flower and growth capacity of the accessions has been observed from the multivariate data analysis.     

An exogenous marker: a novel approach for the characterization of oxidative stress

Oxidative stress (OS) and its consequences which promote alterations in biomolecules, to tissue damage and to the development of pathological conditions, continue to attract many investigators. The identification of reliable biomarker is essential for the characterization of OS and possibly for early discovery of OS-associated diseases. The aim of the present study was to offer a new concept in the development of novel probes for OS, based on the design, synthesis, and utilization of exogenous markers, as alternative to the search for endogenous markers. This article describes: (a) the synthesis of such a marker, linoleoyl tyrosine 2-deoxyguanosyl ester (LTG), constructed from three endogenous subunits: linoleic acid, tyrosine, and 2'-deoxyguanosine, representing the three major groups from which the body is composed, unsaturated fatty acids (USFA), proteins, and DNA, respectively, all bound covalently and (b) the development of analytical tools (LC/MS/MS) to enable the identification of the different LTG oxidized products formed under OS by exposure of LTG to different reactive oxygen species (ROS) such as, copper ions and hypochlorous acid.     

Transgenic sequences are frequently lost in Phytophthora parasitica transformants without reversion of the transgene-induced silenced state

Little data exist on the mechanism and stability of transformation in Phytophthora parasitica, a major oomycete parasite of plants. Here, we studied the stability of drug-resistant protoplast transformants by analyzing single-zoospore derivatives. We show that the transgenic sequences are not stably integrated into the chromosomes, resulting in the loss of drug resistance in single-zoospore derivatives. However, in strains where the P. parasitica gene encoding the CBEL elicitor was silenced by transformation with sense or antisense constructs, silencing is not reversed when the transgenic sequences are lost. This suggests that instability of P. parasitica transformants is not an obstacle for loss-of-function studies in this organism.     

Genetic variation and possible origins of weedy rice found in California

Control of weeds in cultivated crops is a pivotal component in successful crop production allowing higher yield and higher quality. In rice‐growing regions worldwide, weedy rice (Oryza sativa f. spontanea Rosh.) is a weed related to cultivated rice which infests rice fields. With populations across the globe evolving a suite of phenotypic traits characteristic of weeds and of cultivated rice, varying hypotheses exist on the origin of weedy rice. Here, we investigated the genetic diversity and possible origin of weedy rice in California using 98 simple sequence repeat (SSR) markers and an Rc gene‐specific marker. By employing phylogenetic clustering analysis, we show that four to five genetically distinct biotypes of weedy rice exist in California. Analysis of population structure and genetic distance among individuals reveals diverse evolutionary origins of California weedy rice biotypes, with ancestry derived from indica, aus, and japonica cultivated rice as well as possible contributions from weedy rice from the southern United States and wild rice. Because this diverse parentage primarily consists of weedy, wild, and cultivated rice not found in California, most existing weedy rice biotypes likely originated outside California.     

Chalcones as potent tyrosinase inhibitors: the importance of a 2,4-substituted resorcinol moiety

Compounds, which inhibit tyrosinase, could be effective as depigmenting agents. We have introduced a group of mono-, di-, tri- and tetra-substituted hydroxychalcones as effective tyrosinase inhibitors, showing that the most important factor determining tyrosinase inhibition efficiency is the position of the hydroxyl group(s) rather their number. The aim of the present study was to investigate the contribution of the different functional groups of the tetrahydroxychalcones to their inhibitory potency, with a view to optimizing the design of whitening agents. Four tetrahydroxychalcones were evaluated, the commercially available Butein and other three were synthesized, and their inhibitory effect on tyrosinase was tested. Results showed that a 2,4-substituted resorcinol subunit on ring B contributed the most to inhibitory potency. Changing the resorcinol substitute to position 3,5- or placing it on ring A significantly diminished the inhibitory effect of the compounds. A catechol subunit on ring A acted as a metal chelator (in the presence of copper ions) and as a competitive inhibitor (in the presence of tyrosinase), while a catechol on ring B oxidized to o-quinone (in the presence of both copper ions and tyrosinase). Three of the compounds also demonstrated antioxidant activity, which may contribute to the prevention of pigmentation. An examination of correlations between inhibitory activity and physical properties of the chalcones tested (such as dissociation energy and molecular planarity) showed positive correlation with the moment dipole value in the Y-axis, which may be used as an indicator of the inhibitory potential of new molecules. The present study revealed two very active tyrosinase inhibitors, 2,4,3',4'-hydroxychalcone and 2,4,2',4'-hydroxychalcone (with IC50 of 0.2 and 0.02 microM, respectively). Structure-related activity studies added some understanding of the role and contribution of different functional groups associated with tyrosinase inhibitors.     

Acute Short-Term Mental Stress Does Not Influence Salivary Flow Rate Dynamics

Background: Results of studies that address the influence of stress on salivary flow rate and composition are controversial. The aim of this study was to reveal the influence of stress vulnerability and different phases of stress reactivity on the unstimulated and stimulated salivary flow rate. We examined that acute mental stress does not change the salivary flow rate. In addition, we also examined the salivary cortisol and protein level in relation to acute mental stress stimuli. Methods: Saliva of male subjects was collected for five minutes before, immediately, 10, 30 and 120 min after toothbrushing. Before toothbrushing, the subjects were exposed to acute stress in the form of a 2 min public speech. Salivary flow rate and total protein was measured. The physiological stress marker cortisol was analyzed using enzyme-linked immunosorbent assay. To determine the subjects'' psychological stress reaction, the State-Trait-Anxiety Inventory State questionnaire (STAI) data were obtained. The subjects were divided into stress subgroup (S1) (psychological reactivity), stress subgroup (S2) (psychological and physiological reactivity) and a control group. The area under the curve for salivarycortisol concentration and STAI-State scores were calculated. All data underwent statistical analysis using one-way analysis of variance. Results: Immediately after stress exposure, all participants exhibited a psychological stress reaction. Stress exposure did not change the salivary flow rate. Only 69% of the subjects continued to display a physiological stress reaction 20 minutes after the public talk. There was no significant change in the salivary flow rate during the psychological and the physiological stress reaction phases relative to the baseline. Conclusions: Acute stress has no impact on the salivary flow rate; however, there may be other responses through salivary proteins that are increased with the acute stress stimuli. Future studies are needed to examine specific proteins and their possible roles in acute stress responses.     

Epithelial Mesenchymal Transition and Pancreatic Tumor Initiating CD44+/EpCAM+ Cells Are Inhibited by γ-Secretase Inhibitor IX

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease with a high rate of metastasis. Recent studies have indicated that the Notch signalling pathway is important in PDAC initiation and maintenance, although the specific cell biological roles of the pathway remain to be established. Here we sought to examine this question in established pancreatic cancer cell lines using the γ-secretase inhibitor IX (GSI IX) to inactivate Notch. Based on the known roles of Notch in development and stem cell biology, we focused on effects on epithelial mesenchymal transition (EMT) and on pancreatic tumor initiating CD44+/EpCAM+ cells. We analyzed the effect of the GSI IX on growth and epithelial plasticity of human pancreatic cancer cell lines, and on the tumorigenicity of pancreatic tumor initiating CD44+/EpCAM+ cells. Notably, apoptosis was induced after GSI IX treatment and EMT markers were selectively targeted. Furthermore, under GSI IX treatment, decline in the growth of pancreatic tumor initiating CD44+/EpCAM+ cells was observed in vitro and in a xenograft mouse model. This study demonstrates a central role of Notch signalling pathway in pancreatic cancer pathogenesis and identifies an effective approach to inhibit selectively EMT and suppress tumorigenesis by eliminating pancreatic tumor initiating CD44+/EpCAM+ cells.