Strigo-D2-a bio-sensor for monitoring spatio-temporal strigolactone signaling patterns in intact plants.

Strigolactones (SLs) are a class of plant hormones that mediate biotic interactions and modulate developmental programs in response to endogenous and exogenous stimuli. However, a comprehensive view on the spatio-temporal pattern of SL signaling has not been established, and tools for a systematic in planta analysis do not exist. Here, we present Strigo-D2, a genetically encoded ratiometric SL signaling sensor that enables the examination of SL signaling distribution at cellular resolution and is capable of rapid response to altered SL levels in intact Arabidopsis (Arabidopsis thaliana) plants. By monitoring the abundance of a truncated and fluorescently labeled SUPPRESSOR OF MAX2 1-LIKE 6 (SMXL6) protein, a proteolytic target of the SL signaling machinery, we show that all cell types investigated have the capacity to respond to changes in SL levels but with very different dynamics. In particular, SL signaling is pronounced in vascular cells but low in guard cells and the meristematic region of the root. We also show that other hormones leave Strigo-D2 activity unchanged, indicating that initial SL signaling steps work in isolation from other hormonal signaling pathways. The specificity and spatio-temporal resolution of Strigo-D2 underline the value of the sensor for monitoring SL signaling in a broad range of biological contexts with highly instructive analytical depth.

Drought stress modulates cuticular wax composition of the grape berry.

Drought events are a major challenge for many horticultural crops, including grapes, which are often cultivated in dry and warm climates. It is not understood how the cuticle contributes to the grape berry response to water deficit (WD); furthermore, the cuticular waxes and the related biosynthetic pathways are poorly characterized in this fruit. In this study, we identified candidate wax-related genes from the grapevine genome by phylogenetic and transcriptomic analyses. Developmental and stress response expression patterns of these candidates were characterized across pre-existing RNA sequencing data sets and confirmed a high responsiveness of the pathway to environmental stresses. We then characterized the developmental and WD-induced changes in berry cuticular wax composition, and quantified differences in berry transpiration. Cuticular aliphatic wax content was modulated during development and an increase was observed under WD, with wax esters being strongly up-regulated. These compositional changes were related to up-regulated candidate genes of the aliphatic wax biosynthetic pathway, including CER10, CER2, CER3, CER1, CER4, and WSD1. The effect of WD on berry transpiration was not significant. This study indicates that changes in cuticular wax amount and composition are part of the metabolic response of the grape berry to WD, but these changes do not reduce berry transpiration.

Iron Supply Affects Anthocyanin Content and Related Gene Expression in Berries of Vitis vinifera cv. Cabernet Sauvignon.

Anthocyanins are important compounds for red grape and red wine quality, and can be influenced by supply of nutrients such as nitrogen, phosphorus, potassium, zinc, and iron. The present work aims to gain a better understanding of the effect of iron supply on anthocyanins concentration in grape berries. To this end, own-rooted four-year-old Cabernet Sauvignon grapevines (Vitis vinifera) were fertigated every three days with 0, 23, 46, 92, and 184 µM iron (Fe) from ferric ethylenediamine di (o-hydroxyphenylacetic) acid (Fe-EDDHA) in a complete nutrient solution. Fe deficiency or excess generally led to higher concentrations of titratable acidity and skin/berry ratio, and to lower reducing sugar content, sugar/acid ratio, pH, berry weight, and concentration of anthocyanins. Most of the individual anthocyanins detected in this study, except cyanidin-3-O-glucoside, delphinidin-3-O-glucoside, and cyanidin-3-O-(6-O-coumaryl)-glucoside, in moderate Fe treatment (46 µM) grapes were significantly higher than those of other treatments. Genes encoding chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), leucoanthocyanidin dioxygenase (LDOX), and anthocyanin O-methyltransferase (AOMT) exhibited higher transcript levels in berries from plants cultivated with 46 µM Fe compared to the ones cultivated with other Fe concentrations. We suggest that grape sugar content, anthocyanins content, and transcriptions of genes involved in anthocyanin biosynthesis were correlated with Fe supply concentrations.

Nucleocytoplasmic shuttling of valosin-containing protein (VCP/p97) regulated by its N domain and C-terminal region.

Valosin-containing protein (VCP or p97), a member of the AAA family (ATPases associated with diverse cellular activities), plays a key role in many important cellular activities. A genetic deficiency of VCP can cause inclusion body myopathy associated with Paget's disease of bone and frontotemporal dementia (IBMPFD). Previous studies showed that the VCP N domain is essential for the regulation of nuclear entry of VCP. Here we report that IBMPFD mutations, which are mainly located in the N domain, suppress the nuclear entry of VCP. Moreover, the peptide sequence G780AGPSQ in the C-terminal region regulates the retention of VCP in the nucleus. A mutant lacking this sequence can increase the nuclear distribution of IBMPFD VCP, suggesting that this sequence is a potential molecular target for correcting the deficient nucleocytoplasmic shuttling of IBMPFD VCP proteins.

Valosin-containing protein (VCP/p97) is capable of unfolding polyubiquitinated proteins through its ATPase domains.

Valosin-containing protein (VCP or p97) is required for the proteasomal degradation of polyubiquitinated proteins. However, the molecular mechanism for VCP to process the polyubiquitinated proteins remains unclear. Here, we show that VCP can unfold polyubiquitinated proteins. It preferably unfolds the pentaubiquitin-over monoubiquin-conjugated dihydrofolate reductase (Ub5-DHFR or Ub-DHFR) in a dose dependent manner. In addition, the unfolding activity of VCP does not depend on its ATPase activity, on the contrary, ATP and its non-hydrolysable analogs suppress the unfolding of Ub5-DHFR. The structural and functional analysis showed that either D1 or D2 domain of VCP is sufficient to carry out this unfolding activity. The structure of the substrates also affects its unfolding by VCP. VCP is unable to unfold Ub5-DHFR in a tight structure when it binds with methotrexate, a folate analog with high affinity to DHFR. Thus, these results support that VCP is capable of unfolding polyubiquitinated proteins and suggest that VCP may facilitate the proteasomal degradation of polyubiquitinated proteins through its unfolding activity.

Tissue-Specific Expression Analysis of Anthocyanin Biosynthetic Genes in White- and Red-Fleshed Grape Cultivars.

Yan73, a teinturier (dyer) grape variety in China, is one of the few Vitis vinifera cultivars with red-coloured berry flesh. To examine the tissue-specific expression of genes associated with berry colour in Yan73, we analysed the differential accumulation of anthocyanins in the skin and flesh tissues of two red-skinned grape varieties with either red (Yan73) or white flesh (Muscat Hamburg) based on HPLC-MS analysis, as well as the differential expression of 18 anthocyanin biosynthesis genes in both varieties by quantitative RT-PCR. The results revealed that the transcripts of GST, OMT, AM3, CHS3, UFGT, MYBA1, F3'5'H, F3H1 and LDOX were barely detectable in the white flesh of Muscat Hamburg. In particular, GST, OMT, AM3, CHS3 and F3H1 showed approximately 50-fold downregulation in the white flesh of Muscat Hamburg compared to the red flesh of Yan73. A correlation analysis between the accumulation of different types of anthocyanins and gene expression indicated that the cumulative expression of GST, F3'5'H, LDOX and MYBA1 was more closely associated with the acylated anthocyanins and the 3'5'-OH anthocyanins, while OMT and AM3 were more closely associated with the total anthocyanins and methoxylated anthocyanins. Therefore, the transcripts of OMT, AM3, GST, F3'5'H, LDOX and MYBA1 explained most of the variation in the amount and composition of anthocyanins in skin and flesh of Yan73. The data suggest that the specific localization of anthocyanins in the flesh tissue of Yan73 is most likely due to the tissue-specific expression of OMT, AM3, GST, F3'5'H, LDOX and MYBA1 in the flesh.

Effect of Three Training Systems on Grapes in a Wet Region of China: Yield, Incidence of Disease and Anthocyanin Compositions of Vitis vinifera cv. Cabernet Sauvignon.

Grapevine training systems determine the suitability for grape varieties in a specific growing region. We evaluated the influence of three training systems, Single Guyot (SG), Spur-pruned Vertical Shoot-Positioned (VSP), and Four-Arm Kniffin (4AK), on the performance of grapes and vines of Vitis vinifera L. cv. Cabernet Sauvignon in the 2012 and 2013 growing seasons in a wet region of central China. 4AK was the most productive system in comparison to SG and VSP. SG and VSP had lower disease infections of leaves and berries, especially in the mid- and final stage of berry ripening. Three training systems had no impact on berry maturity. PLS-DA (Partial Least Squares-Discriminant) analysis showed that the relatively dry vintage could well discriminate three training systems, but the wet vintage was not. A wet vintage of 2013 had more accumulation of 3'5'-substituted and acylated anthocyanins, including malvidin-3-O-(6-O-acetyl)-glucoside, malvidin-3-O-glucoside, and petunidin-3-O-(cis-6-O-coumaryl)-glucoside, etc. With regard to the effect of training systems, 4AK grapes had the lowest concentrations of total anthocyanins and individual anthocyanins, SG and VSP differed according to the different vintages, and showed highest concentration of total individual anthocyanins in 2012 and 2013, respectively. Generally, VSP benefited the most, contributing to significantly highest levels of total individual anthocyanins, and major anthocyanin, including malvidin-3-O-glucoside and malvidin-3-O-(6-O-acetyl)-glucoside, and the grapes obtained from VSP presented significantly highest proportion of 3'5'-substituted anthocyanins. With regard to the ratios of 3'5'/3'-substituted, methoxylated/non-methoxylated and acylated/non-acylated anthocyanins, the significantly higher levels were also shown in VSP system. In summary, VSP was the best training system for Cabernet Sauvignon to accumulate relatively stable individual anthocyanins in this wet region of China and potentially in other rainy regions.

Promoting effect of foliage sprayed zinc sulfate on accumulation of sugar and phenolics in berries of Vitis vinifera cv. Merlot growing on zinc deficient soil.

The effect of foliage sprayed zinc sulfate on berry development of Vitis vinifera cv. Merlot growing on arid zone Zn-deficient soils was investigated over two consecutive seasons, 2013 and 2014. Initial zinc concentration in soil and vines, photosynthesis at three berry developmental stages, berry weight, content of total soluble solids, titratable acidity, phenolics and expression of phenolics biosynthetic pathway genes throughout the stages were measured. Foliage sprayed zinc sulfate showed promoting effects on photosynthesis and berry development of vines and the promotion mainly occurred from veraison to maturation. Zn treatments enhanced the accumulation of total soluble solids, total phenols, flavonoids, flavanols, tannins and anthocyanins in berry skin, decreasing the concentration of titratable acidity. Furthermore, foliage sprayed zinc sulfate could significantly influence the expression of phenolics biosynthetic pathway genes throughout berry development, and the results of expression analysis supported the promotion of Zn treatments on phenolics accumulation. This research is the first comprehensive and detailed study about the effect of foliage sprayed Zn fertilizer on grape berry development, phenolics accumulation and gene expression in berry skin, providing a basis for improving the quality of grape and wine in Zn-deficient areas.

The protective role of antifreeze protein 3 on the structure and function of mature mouse oocytes in vitrification.

Several studies have reported the oocyte damage in mice during vitrification; however, little has been known about the protective role that antifreeze protein 3 (Afp3) plays on their cellular structure and function during vitrification. In order to observe the extracellular cryo-protective role of Afp3, four groups were divided randomly. The observations were made for changes in cytoskeleton, expression of the related genes before and after vitrification, and also for changes in the in vitro developmental potential of oocytes. The outcomes were as follows: (i) microtubules, actin filaments and chromosomal integrity were more intact in the vitrification group supplemented with additional Afp3 compared to the vitrification group. In the fresh control group and the group with additional cryoprotectant containing ethylene glycol (EG), dimethyl sulfoxide (Me2SO) and sucrose, the organelles were more intact than the other two vitrification groups. (ii) Real-time PCR analysis revealed that the relative quantification of mitotic arrest deficient 2 (Mad2) and centromere protein E (Cenp-e) were significantly higher in the vitrification group with additional Afp3, the fresh control group and the one group with additional cryoprotectant, in comparison to the vitrification group. On the contrary, the expression of cold inducible RNA-binding protein (Cirbp) and kinesin-5 motor protein (Eg5) were up-regulated in the vitrification group compared to the remaining groups. (iii) The fertilization rate and the recovery rate in the fresh control group and the group with additional cryoprotectant were higher than the other two vitrification groups; furthermore, the recovery rate and the fertilization rate in the vitrification group with Afp3 were higher than the vitrification group. However, the blastocyst formation rate in all the four groups showed no statistical significance. In conclusion, Afp3 plays a positive role in the structure and function of mice oocytes in vitrification.

Hemorphin-Based Analgesia: A Mechanism of Cupping Technique?

Background and Objective: Cupping is a time-honoured traditional healing modality for pain management and remains favoured by professionals and lay people across several cultures today. However, the analgesic mechanism of cupping is still poorly understood. In addition, clinical guidelines for standardized applications of cupping are currently lacking. The awareness of cupping marks has provoked curiosity about the connection between skin color changes and their benefit for local pain relief. Computer simulation is a promising approach for numerical modeling the cupping-evoked erythrocyte emigration. Quantitative proteomic profiling of cupping-induced blister fluid exhibited a significant decrease in the abundance of haemoglobin ß subunit. This finding provides a critical clue to paint a novel picture of the mechanism behind cupping. The hemorphins are a set of non-classical opioid peptides derived from the proteolysis of haemoglobin ß subunit. In the present study, a probable mechanism of hemorphin-based cupping analgesia is proposed. The hemorphin could also act as a potential biomarker for objective and timely quantitative clinical assessment of cupping in the management of pain conditions. A seminal theory may open a new avenue for future translational research on promoting the efficacy and safety of cupping analgesia. Conclusion: The local analgesic effect of cupping is probable in the context of haemoglobin degradation that bestows the appearance of hemorphins along with engaging opioid receptor signalling. Exploring the potential novel mechanism of cupping analgesia facilitates seeking non-pharmacologic pain interventions.

GreenGate 2.0: Backwards compatible addons for assembly of complex transcriptional units and their stacking with GreenGate.

Molecular cloning is a crucial technique in genetic engineering that enables the precise design of synthetic transcriptional units (TUs) and the manipulation of genomes. GreenGate and several other modular molecular cloning systems were developed about ten years ago and are widely used in plant research. All these systems define grammars for assembling transcriptional units from building blocks, cloned as Level 0 modules flanked by four-base pair overhangs and recognition sites for a particular Type IIs endonuclease. Modules are efficiently assembled into Level 1 TUs in a hierarchical assembly process, and Level 2 multigene constructs are assembled by stacking Level 1 TUs. GreenGate is highly popular but has three main limitations. First, using ad-hoc overhangs added by PCR and classical restriction/ligation prevents the efficient use of a one-pot, one-step reaction to generate entry clones and domesticate internal sites; second, a Level 1 TU is assembled from a maximum of six modules, which may be limiting for applications such as multiplex genome editing; third, the generation of Level 2 assemblies is sequential and inefficient. GreenGate 2.0 (GG2.0) expands GreenGate features. It introduces additional overhangs, allowing for the combination of up to 12 Level 0 modules in a Level 1 TU. It includes a Universal Entry Generator plasmid (pUEG) to streamline the generation of Level 0 modules. GG2.0 introduces GreenBraid, a convenient method for stacking transcriptional units iteratively for multigene assemblies. Importantly, GG2.0 is backwards compatible with most existing GreenGate modules. Additionally, GG2.0 includes Level 0 modules for multiplex expression of guide RNAs for CRISPR/Cas9 genome editing and pre-assembled Level 1 vectors for dexamethasone-inducible gene expression and ubiquitous expression of plasma membrane and nuclear fluorescent markers. GG2.0 streamlines and increases the versatility of assembling complex transcriptional units and their combination.

Day Temperature Has a Stronger Effect Than Night Temperature on Anthocyanin and Flavonol Accumulation in 'Merlot' (Vitis vinifera L.) Grapes During Ripening.

Flavonoids impart color and mouthfeel to grapes and wine and are very sensitive to environmental conditions. Growth chamber experiments were performed to investigate the effect of temperature regimes and the differences between day/night temperatures on anthocyanins and flavonols in Merlot grapes. Among the regimes tested, the ones with diurnal 20°C determined the highest levels of anthocyanins and flavonols. Higher diurnal temperatures decreased those levels but increased the proportion of methoxylated and acylated species. When regimes with the same day temperature but different night temperatures were compared, differences between day/night temperatures did not affect anthocyanins, unless a difference of 25°C between day and night temperatures was imposed. When regimes with the same night temperature but different day temperatures were compared, the regime with higher day temperature had a lower anthocyanin level. No relationships were observed between the effects of temperature regimes on anthocyanin level and the expression of key anthocyanin genes. However, the effects on anthocyanin acylation level were consistent with the effects on the acyltransferase expression, and the effects on flavonol level were consistent with the effects on the expression of key flavonol genes. This study indicates that, in Merlot grapes, anthocyanins and flavonols are mostly sensitive to day temperatures.

Dynamic changes in anthocyanin biosynthesis regulation of Cabernet Sauvignon (Vitis vinifera L.) grown during the rainy season under rain-shelter cultivation.

The grapevine (Vitis vinifera L.) berry coloring mechanism in response to seasonal rain during grape ripening remains poorly understood. Therefore, anthocyanin biosynthesis regulation, dynamic changes in anthocyanin accumulation, biosynthetic enzyme activities, and related gene expression patterns were investigated in Cabernet Sauvignon grown under rain-shelter cultivation and open-field cultivation. Results showed that anthocyanin biosynthesis was strongly repressed during the rainy season. Environmental fluctuation from seasonal rain provoked metabolic responses in grapes, and there was a significantly greater accumulation of most of the anthocyanins, mainly the compositions of non-acylated and non-methylated, under rain-shelter cultivation; these findings indicate that rain-shelter cultivation may help improve tolerance to seasonal rain-induced stresses. Obvious resilience was observed in anthocyanins of open-field-cultivated grapes at harvest. Hierarchical cluster analysis indicated strong correlations between anthocyanin contents, CHI and DFR activities, and VvMYB5b transcriptional level. These findings provide novel insight into the crucial factors that directly modulate anthocyanin biosynthesis and consequently control grape coloration.

ABA Alleviates Uptake and Accumulation of Zinc in Grapevine (Vitis vinifera L.) by Inducing Expression of ZIP and Detoxification-Related Genes.

Abscisic acid (ABA) is a plant hormone that can mitigate heavy metal toxicity. Exogenous ABA and ABA mimic 1 (AM1) were applied to study the influence on Zn uptake and accumulation in Vitis vinifera L. cv. Merlot seedlings exposed to excess Zn. The seedlings were treated with either normal or excess levels of Zn in combination with applications of ABA and AM1. Excess Zn exposure resulted in decreased lateral root length, decreased photosynthesis, elevated uptake, and accumulation of Zn in roots, trunks, and stems, decreased jasmonic acid content in roots and leaves, and induced the expression of Zn transportation- and detoxification-related genes. Remarkably, in the presence of toxic amounts of Zn, the exogenous application of ABA, but not of AM1, reduced the uptake and accumulation of Zn in roots and induced higher expression of both ZIP genes and detoxification-related genes in root and leaf. These results indicate that exogenous ABA enhances the tolerance of grape seedlings to excess Zn and that AM1 is not a suitable ABA mimic compound for Zn stress alleviation in grapes.

Foliar applications of iron promote flavonoids accumulation in grape berry of Vitis vinifera cv. Merlot grown in the iron deficiency soil.

Flavonoids are important compounds for grape and wine quality. Foliar fertilization with iron compounds has been reported to have a substantial impact on grape composition in the grapevines growing in calcareous soil. However, much less is known about its real impact on flavonoid composition. In the present study, Ferric ethylenediamine di (O-hydroxyphenylacetic) acid (Fe-EDDHA) was foliar applied to Merlot (Vitis vinifera L.) grapevines growing in calcareous soil over two consecutive vintages in order to study its effect on grape flavonoid composition. Fe-EDDHA foliar supply tended to increase grape sugar, anthocyanin and flavonol content, decrease acid content and enhance the juice pH when compared to the control. Principal component analysis showed that the vintage also had influence on grape quality. The results suggested that Fe-EDDHA foliar application had an enhancement effect on grape secondary metabolism, and the effect increased the nutritional value of the consequent grapes and wines.

Methoxypyrazines biosynthesis and metabolism in grape: A review.

This review summarizes research on the discovery, biosynthesis, accumulation, transport, and metabolism of 3-alkyl-2-methoxypyrazines (MPs) in grape. The MPs are a family of potent volatile compounds distributed throughout biological kingdoms. These compounds impart herbaceous/green/vegetal sensory attributes to certain varieties of wine. Generally, high levels of MPs in wine are derived mainly from the corresponding grapes. Although two pathways for MPs biosynthesis have been proposed, only the final step and the enzymes that catalyze it has been confirmed in grape, and the metabolic intermediates and key enzymes involved in other steps are still unknown. The limited understanding of MPs metabolism has restricted research on these compounds, and some empirical results cannot be explained by the current knowledge of MPs metabolism. This review provides insights into research on MPs biosynthesis and metabolism, and proposes directions for further research on this important class of flavour/odour compounds.

Influence of natural variation in berry size on the volatile profiles of Vitis vinifera L. cv. Merlot and Cabernet Gernischt grapes.

This study was conducted during the 2014 and 2015 vintages on Vitis vinifera L. cv. Merlot and Cabernet Gernischt to investigate whether natural variation in berry size could affect grape aromatic compounds. Grape berries were separated into three size categories based on their diameter: small, middle and large. The results showed that berry size exerted a significant influence on the volatile profiles of both winegrape varieties. Hierarchical clustering analysis demonstrated that the volatile profiles of middle berries were different from those of large and small berries. Middle berries had the greatest abundance of aroma compounds, followed by small and large berries. Especially, C6/C9 compounds, norisoprenoids, terpenoids showed markedly different concentrations among differently sized Merlot berries and C6/C9 compounds, terpenoids among differently sized Cabernet Gernischt berries. Middle berries of both grape varieties may possess the greatest intensity of fresh-green, fruity and floral aromas due to the high odour activity values (OAVs) of decanal, hexanal, (E)-2-hexenal, (E)-ß-damascenone and ß-ionone in middle sizes of Merlot berries and the high OAVs of (E)-2-hexenal and (E)-ß-damascenone in middle sizes of Cabernet Gernischt berries. This knowledge could be important for winemakers to conduct targeted berry sorting, thereby improving the aromatic quality of grapes.

[Azoospermia factor microdeletions in idiopathic azoospermia and severe oligozoospermia].

OBJECTIVE: To observe the relationship between microdeletions of AZF( azoospermia factor) on Y chromosome in male with idiopathic azoospermia and severe oligozoospermia. METHODS: Only patients with an apparently normal 46,XY karyotype and normal FSH, LH and T were included in this study. Multiplex PCR was used to detect the sequence-tagged sites( STS) as follows :sY84, sY86, sY127, sY134, sY152, sY153, sY254, sY255, and ZFX/Y was used as internal control gene. RESULTS: No microdeletion was detected in the control whereas 8 microdeletion cases existed in 67 idiopathic azoospermia and severe oligozoospermia, including 4 in AZFc, 2 in AZFa + AZFc, 1 in AZFc + AZFb, and 1 in AZFb. The prevalence rate of microdeletion was 11.94%, which was statistically different from the control. CONCLUSION: Microdeletions in the AZF regions on the long arm of the Y-chromosome are associated with idiopathic azoospermic and severely oligozoospermic men. Multiplex PCR was a rapid and reliable method for screening microdeletions of AZF.

Effect of NS398 on metastasis-associated gene expression in a human colon cancer cell line.

AIM: To investigate the effect of NS398 on the metastasis-associated gene expression in LoVo colorectal cancer cells. METHODS: LoVo cells were treated with NS398 at the concentration of 100 micromol/L for 24 and 48 h respectively. Total RNA was extracted with TRIzol reagents and reverse transcribed with Superscript II and hybridized with cDNA microarray (containing oncogenes, tumor suppressor genes, signal transduction molecules, adhesive molecules, growth factors, and ESTs) fabricated in our laboratory. After normalization, the ratio of gene expression of NS398 treated to untreated LoVo cells was either 2-fold up or 0.5-fold down was defined as the differentially expressed genes. Semi-quantitative RT-PCR was used to validate the microarray results. RESULTS: Among the 447 metastasis-associated genes, 9 genes were upregulated and 8 genes were downregulated in LoVo cells treated with NS398 for 24 h compared to untreated cells. While 31 genes were upregulated and 14 genes were downregulated in LoVo cells treated with NS398 for 48 h. IGFBP-5, PAI-2, JUN, REL, BRCA1, and BRCA2 might be the new targets of NS398 in treatment of colorectal cancer. CONCLUSION: NS398 might exert its anti-metastasis effect on colorectal cancer by affecting several metastasis-associated gene expression.

Melatonin treatment of pre-veraison grape berries to increase size and synchronicity of berries and modify wine aroma components.

A comprehensive investigation was carried out to determine the effect of exogenous melatonin treatment of pre-veraison grapes on grape berries and its wines. Two melatonin treatments of pre-veraison grape berries increased the weight of the berries by approximately 6.6%. Meanwhile, this melatonin treatment could be beneficial in the reduction of underripe and overripe fruits and in enhancing the synchronicity of the berries. In addition, there were significant differences in the volatile compound composition between the wine produced from the melatonin-treated berries and the wines made from untreated berries. The wine from melatonin-treated pre-veraison grape berries had stronger fruity, spicy, and sweet sensory properties, compared to the wines made from untreated berries. Prolonging the treatment through repeated applications can enhance these effects and under different seasonal conditions, more pronounced effects on the grape quality and wine properties can be observed.