Our previous study revealed stem inclusion fermentation reduced anthocyanin, and increased tannin and aroma compounds responsible for green notes. This study further investigated the effect of clone selection and whole bunch fermentation on Pinot noir wine composition, with focus on tannin composition. Three treatments were conducted using two clones (AM10/5 and UCD5) in 2021 and 2022: 100% destemmed (DS), 30% whole bunch (WB30), and 60% whole bunch (WB60). WB60 increased stem and skin derived tannins but reduced seed derived tannin proportion in wines. Clone selection had an impact on tannin composition and an even greater impact on tannin concentration, colour, and aroma compounds. AM10/5 produced wines with higher tannin, polymeric pigments and darker colour. AM10/5 wines also had higher concentration of phenylethyl alcohol, but lower concentrations of 3-isobutyl-2-methoxypyrazine and ethyl esters, indicating more floral but less fruity and green notes.

This study aimed to investigate the impact of leaf removal on concentrations of anthocyanin, tannin, and methoxypyrazines (MPs) in Pinot noir grapes and wines. Leaf removal after 7 days (LR7), 30 days (LR30), and 60 days (LR60) of flowering were compared with no leaf removal control (LRC). Grapes and resultant wines were analysed for tannin and aroma composition using liquid chromatography and two-dimensional gas chromatography-mass spectrometry. All leaf removal treatments increased anthocyanin concentration in grapes and reduced MP levels in grape stems compared to LRC, indicating the effectiveness of both early and late leaf removal. Leaf removal after 7 days and 30 days were more effective in enhancing colour density, polymeric pigments, and tannin concentration in wines. Higher grape skin tannin and anthocyanin concentrations, along with lower seed tannin concentration in berries, correlated with higher tannin concentrations in wines. LR7 showed significantly higher skin-originated tannin proportion than LRC, suggesting a useful tool to manage tannin extraction. Aroma composition of resultant wines was influenced by leaf removal, although these differences were not evident in the sensory evaluation.

Ladybirds (Coccinellidae) use toxic compounds, mostly alkaloids in their haemolymph, for defence against predators and other enemies. The toxicity of ladybirds to predators cannot be directly assessed because predators show avoidance reactions without ingesting the beetles. The alkaloid of ladybird Harmonia axyridis showed wide range toxicity to diverse non-target organisms. Thus, we used a quick, inexpensive and easy-to-perform method using bioassays on water flea Daphnia magna for comparative quantification of the toxicity (LD50) of whole body extracts from several species of ladybirds that differ in their warning colouration. Alien invasive aposematic polymorphic ladybird H. axyridis was more toxic than all the other species examined: aposematic Adalia bipunctata > cryptic Cynegetis impunctata > aposematic Coccinella septempunctata > slightly aposematic Calvia quatuordecimguttata. Three month old adults of H. axyridis were 3.8 times more toxic than two week and one month old adults. The two most common colour morphs (non-melanic novemdecimsignata and melanic spectabilis) did not differ in their toxicity. High toxicity of H. axyridis as compared to all other species examined may contribute to the invasiveness of this species.

Two Pinot noir clones (AM10/5 and UCD5) were analyzed for tannin and methoxypyrazines (MPs) in different grape tissues during berry development using liquid chromatography and two-dimensional gas chromatography-mass spectrometry. On a per berry basis, skin tannins reached the maximum level about 2-3 weeks after véraison, seed tannins at around véraison, and stem tannins 4 weeks before véraison. Clone AM10/5 showed significantly higher levels of seed and stem tannins on a per berry basis at harvest. Tannin concentration and composition varied among the different tissues. On a per berry basis, stem tannin levels were comparable to skin tannins but were 3 to 4 times lower than seed tannins, while stem tannins had an intermediate galloylation (5-7%) between seed tannins (12-18%) and skin tannins (2%) and lower prodelphinidin (4-7%) than skin tannins (31-36%). The mean degree of polymerization of stem tannins was similar to seed tannins but lower than skin tannins. MPs, including 3-isopropyl-2-methoxypyrazine (IPMP), 3-s-isobutyl-2-methoxypyrazine (SBMP), and 3-isobutyl-2-methoxypyrazine (IBMP), showed significantly higher concentrations than their sensory thresholds in grape stems but not in skins. The MPs development in stems showed an increasing trend toward véraison and then a decreasing trend toward harvest. Compared to AM 10/5, UCD5 stems showed a higher level of MPs, especially significantly higher concentrations of IPMP and IBMP at harvest. The extraction of MPs from grape stems could contribute negative green and vegetative characters to Pinot noir wines.

Methoxypyrazines (MPs) are a unique class of volatile compounds containing nitrogenous heterocyclics that impart green bell, vegetal and herbal odors to red grape berries and wines. In this study, the quality and MPs levels of grape berries from six representative red wine grape varieties were determined in the two consecutive years. The results showed that, at maturity, the highest total soluble solid was observed in Petit Verdot grape berries in the two consecutive years. While the anthocyanin content showed the highest in Marselan berries in 2018, in 2019, Petit Verdot berries had the highest anthocyanin content. Moreover, 2-methoxypyrazine (MOMP), 3-methyl-2-methoxypyrazine (MEMP) and 3-ethyl-2-methoxypyrazine (ETMP) levels were relatively lower, with almost no detectable in berries at maturity. The relative higher 3-isobutyl-2-methoxypyrazine (IBMP) content was observed in Cabernet Sauvignon, Marselan, Merlot, and Malbec berries. However, 3-sec-butyl-2-methoxypyrazine (SBMP) and IBMP were only detected in six wines, and their levels were higher than those in the grape berries. Furthermore, correlation analysis showed that there was a statistically positive correlation between the expression levels of VvOMT1 and VvOMT3 and MPs content in grape berries, while the lowest association was found in the VvOMT2. These findings provide a basis for selecting the most suitable grape varieties to improve wine quality.

Cardiovascular disease (CVD) remains the major cause of death worldwide, accounting for almost 31% of the global mortality annually. Several preclinical studies have indicated that ginseng and the major bioactive ingredient (ginsenosides) can modulate several CVDs through diverse mechanisms. However, there is paucity in the translation of such experiments into clinical arena for cardiovascular ailments due to lack of conclusive specific pathways through which these activities are initiated and lack of larger, long-term well-structured clinical trials. Therefore, this review elaborates on current pharmacological effects of ginseng and ginsenosides in the cardiovascular system and provides some insights into the safety, toxicity, and synergistic effects in human trials. The review concludes that before ginseng, ginsenosides and their preparations could be utilized in the clinical treatment of CVDs, there should be more preclinical studies in larger animals (like the guinea pig, rabbit, dog, and monkey) to find the specific dosages, address the toxicity, safety and synergistic effects with other conventional drugs. This could lead to the initiation of large-scale, long-term well-structured randomized, and placebo-controlled clinical trials to test whether treatment is effective for a longer period and test the efficacy against other conventional therapies.

The goal of this study is to identify Coffea arabica O-methyltransferase (OMT) genes involved in the biosynthesis of methoxypyrazines. High levels of 2-isopropyl-3-methoxypyrazine (IPMP) and 2-isobutyl-3-methoxypyrazine (IBMP) in coffee beans are associated with the potato taste defect (PTD). Among the 34 putative O-methyltransferase genes identified in the published genome of C. canephora, three genes are highly homologous to known hydroxypyrazine OMT genes. Genes of interest were amplified and sequenced from genomic DNA of single C. arabica beans grown in eight different locations, including regions with endemic PTD. Although C. arabica OMT target sequences were almost identical regardless of source location, individual beans shared numerous polymorphisms in each of the target genes. Two of the predicted C. arabica OMT enzymes were successfully expressed in Escherichia coli and purified, and one enzyme shows slow yet measurable turnover of both 3-isobutyl-2-hydroxypyrazine (IBHP) and 3-isopropyl-2- hydroxypyrazine (IPHP), supporting a possible role of the coffee plant in PTD.

Three 2-methoxy-3-alkylpyrazines (MPs) in Japanese red wine and grape samples were determined by GC-EIMS, using 2-methyl-3-n-propylpyrazine as an internal standard. MPs in the Cabernet Sauvignon red wines were derived not only from the pulp but also from other parts of the grape berries. All of the Cabernet Sauvignon red wines made annually from 1975 to 1994 contained 2-methoxy-3-isobutylpyrazine (isobutylMP), although those made from well-ripened grapes had a low isobutylMP level. It is suggested that the climatic conditions of September might effect the isobutylMP level of Cabernet Sauvignon grapes and red wines. The mean isobutylMP level of Cabernet Sauvignon, Cabernet Franc, and Merlot Japanese commercial red wines was significantly higher than the mean isobutylMP level of Muscat Bailey A and Zweigeltrebe.

Methoxypyrazines are present in the grapes of certain Vitis vinifera varieties including Sauvignon blanc and contribute herbaceous/green aromas to wine. Environmental factors such as light exposure and temperature can influence methoxypyrazine levels, and viticultural interventions such as canopy manipulation have the ability to reduce methoxypyrazine accumulation in grapes. We assessed methoxypyrazine levels and showed that leaf removal significantly reduces accumulation in Sauvignon blanc grapes. The main effect of reducing methoxypyrazines was preveraison, as postveraison treatments had no effect on concentrations at harvest. Methoxypyrazine concentrations in controls peaked preveraison and decreased through harvest. Dilution due to an increase in berry weight was found to be the major driver of decreasing concentrations, as methoxypyrazine levels on a per berry basis were found to increase through development in two of three seasons. In the one year of our study that showed contrasting results, analyses of weather data indicate that warmer than average temperatures appear to be the principal factor affecting the berries\' ability to accumulate and retain methoxypyrazines. We also explored the expression of potential biosynthetic O-methyltransferase genes VvOMT1, VvOMT2, and VvOMT3; no significant differences were observed with respect to effect of leaf removal and light exposure.

MPs (3-alkyl-2-methoxypyrazines) are grape-derived aroma compounds that are associated with detrimental herbaceous flavours in some wines. It is well known that several viticultural and environmental parameters can modulate MP concentrations in grapes, although comprehensive molecular studies have not been conducted in this field. Although the biosynthesis pathway of MPs has not been fully elucidated, four Vitis vinifera O-methyltransferase genes (VvOMT1-4) have been related to be involved in MP biosynthesis. We assessed whether different abiotic stresses induction have an impact on MP levels in grapes and wines from seeded and parthenocarpic fruits. Our results show that the timing of VvOMT3 expression is associated with the period of MPs accumulation in seeded fruits during both abiotic stresses, whereas no association was found in parthenocarpic fruits. These results are discussed in the context of how different viticultural practices can modulate VvOMT gene expression, which has a direct impact on MPs levels in wines.