Impact of the Post-Harvest Period on the Chemical and Sensorial Properties of planifolia and pompona Vanillas.

Vanilla production in Guadeloupe is expanding. The main species grown is Vanilla planifolia, but other species such as Vanilla pompona are also present and required by industries. To upgrade the value of vanilla production on this Caribbean Island, this study was performed to evaluate the aromatic specifies of these vanilla species according to the length of the post-harvest period (2 months and 9 months). For this purpose, Vanilla planifolia and Vanilla pompona were compared through scald and scarification transformation processes, as well as two different refining times (T1 and T2). For chemical characterization, 0.1 g of vanilla bean seeds was used for SMPE/GC-MS measurements, while 0.05 g of vanilla samples was subjected to infusion in milk (0.15%) for sensory evaluation. The latter involved generation of terms of aroma through olfaction and gustation sessions. The chemical results showed a significant difference between the two species, where vanillin was mostly present in Vanilla planifolia, unlike Vanilla pompona, where it was mainly rich in 4-methoxybenzyl alcohol. Interestingly, the second refining time was characterized by the appearance of two major components, 1,3-octadien and acetic acid. For sensory analysis, all the vanillas exhibited a high diversity of aromas including "sweet", "gourmand", "spicy" flavors and so on. The application of factorial correspondence analysis (FAC) as well as the agglomerative hierarchical clustering (AHC) showed differences between the vanilla samples according to both the species and refining time. The combination of these analyses makes it possible to establish a chemical and organoleptic profile of vanillas. Varietal and processing factors both have a major impact on the aroma profile of vanillas.

Large-Scale Micropropagation of Vanilla (Vanilla planifolia Jacks.) in a Temporary Immersion Bioreactor (TIB).

The cultivation of vanilla (Vanilla planifolia) is of economic interest because vanillin is extracted from the fruits of this species. Vanillin is a natural flavoring highly valued in the food market. However, there is a short supply of propagules available for establishing commercial plantations and good-quality plants with phytosanitary certification. Plant tissue culture represents a viable option to supply large amounts of healthy plants to vanilla producers. In addition, the use of temporary immersion systems will allow commercial scale-up and the establishment of biofactories dedicated to in vitro vanilla propagation. This chapter describes a large-scale micropropagation protocol for vanilla using temporary immersion bioreactors (TIB).

Vanilla odor promotes oral feeding in premature infants-A randomized controlled trial.

PROPOSE: Introducing early oral feeding in premature infants is important because it supports intestinal maturation and helps prevent infections. In addition, early oral feeding is likely to contribute to improved neurocognitive outcomes in preterm infants. Several holistic therapeutic strategies have been developed to improve feeding skills, food tolerance, and the ability to drink independently, including practices such as early breastfeeding, oral stimulation, and subsequent olfactory stimulation. Based on several studies using olfactory stimulation with food odors (vanilla, breast milk) to promote oral feeding in preterm infants this study was conducted to test the following hypothesis: Does olfactory stimulation with vanilla or milk odor (breast milk or formula) lead to a reduction in the time required for nasogastric tube weaning in premature infants older than 26 + 6 weeks of gestational age? In addition, does it influence secondary outcomes such as length of hospital stay, weight development, and attainment of greater amounts of independently consumed food? METHODS: Premature with complete or partial feeding by gastric tube and without ventilation were included. For this study, 207 infants over 26 + 6 gestational weeks were randomized into three different study groups. Before each feeding, an olfactory presentation was made with milk odor, a vanilla Sniffin' Stick, or a control stick. In the final analysis, 165 infants were included (87 males, 78 females). At the time of randomization, infants were on average 12 ± 9.5 days old. RESULTS: While the influence of vanilla and milk odor did not provide a significant difference from the control for the primary outcome, a secondary analysis showed a significant group difference in the cumulative amount of independently drunk food consumed in the first ten days was the highest amount in the vanilla group. This time period was chosen due to the high dropout rate after the first ten days. In addition, there was a promising significance for earlier hospital discharge for prematurely below 32 weeks of gestation receiving vanilla odor stimulation in comparison to milk odor stimulation. CONCLUSION: Although the primary outcome of this study (gastric tube removal) did not provide significant results, a significant benefit of vanilla olfactory stimulation for preterm infants was demonstrated in subgroup analysis above milk odor stimulation. Younger preterm infants seem to benefit from the stimulation.

Identification of a ß-Carboline Alkaloid from Chemoselectively Derived Vanilla Bean Extract and Its Prevention of Lipid Droplet Accumulation in Human Hepatocytes (HepG2).

Targeting bioactive compounds to prevent lipid droplet accumulation in the liver, we explored an antioxidative extract from vanilla bean (Vainilla planifolia) after chemo-selective derivatization through heating and acid modification. The chemical analysis of vanilla bean extract through chemoselective derivatization resulted in the identification of sixteen compounds (34-50) using LC-MS/MS analysis. A ß-carboline alkaloid with a piperidine C-ring and a vanillin moiety at C-1 (34) was identified by molecular networking and diagnostic fragmentation filtering approaches. ß-carboline alkaloid 34 exhibited significant inhibitory activity of lipid droplet accumulation (LDAI) in oleic acid-loaded hepatocellular carcinoma HepG2 cells. The LDAI activity was associated with both activation of lipolysis and suppression of lipogenesis in the cells. The study indicates that crude plant extracts, following chemoselective derivatization, may contain bioactive compounds that could be beneficial in preventing hepatosteatosis and could serve as a source of lead compounds for drug development. This approach may be useful to investigate other mixtures of natural products and food resources.

A High-Resolution Network with Strip Attention for Retinal Vessel Segmentation.

Accurate segmentation of retinal vessels is an essential prerequisite for the subsequent analysis of fundus images. Recently, a number of methods based on deep learning have been proposed and shown to demonstrate promising segmentation performance, especially U-Net and its variants. However, tiny vessels and low-contrast vessels are hard to detect due to the issues of a loss of spatial details caused by consecutive down-sample operations and inadequate fusion of multi-level features caused by vanilla skip connections. To address these issues and enhance the segmentation precision of retinal vessels, we propose a novel high-resolution network with strip attention. Instead of the U-Net-shaped architecture, the proposed network follows an HRNet-shaped architecture as the basic network, learning high-resolution representations throughout the training process. In addition, a strip attention module including a horizontal attention mechanism and a vertical attention mechanism is designed to obtain long-range dependencies in the horizontal and vertical directions by calculating the similarity between each pixel and all pixels in the same row and the same column, respectively. For effective multi-layer feature fusion, we incorporate the strip attention module into the basic network to dynamically guide adjacent hierarchical features. Experimental results on the DRIVE and STARE datasets show that the proposed method can extract more tiny vessels and low-contrast vessels compared with existing mainstream methods, achieving accuracies of 96.16% and 97.08% and sensitivities of 82.68% and 89.36%, respectively. The proposed method has the potential to aid in the analysis of fundus images.

Enzymatic Activity and Its Relationships with the Total Phenolic Content and Color Change in the High Hydrostatic Pressure-Assisted Curing of Vanilla Bean (Vanilla planifolia).

Diverse enzymatic reactions taking place after the killing of green vanilla beans are involved in the flavor and color development of the cured beans. The effects of high hydrostatic pressure (HHP) at 50-400 MPa/5 min and blanching as vanilla killing methods were evaluated on the total phenolic content (TPC), polyphenoloxidase (PPO), and peroxidase (POD) activity and the color change at different curing cycles of sweating-drying (C0-C20) of vanilla beans. The rate constants describing the above parameters during the curing cycles were also obtained. The TPC increased from C1 to C6 compared with the untreated green beans after which it started to decrease. The 400 MPa samples showed the highest rate of phenolic increase. Immediately after the killing (C0), the highest increase in PPO activity was observed at 50 MPa (46%), whereas for POD it was at 400 MPa (25%). Both enzymes showed the maximum activity at C1, after which the activity started to decrease. As expected, the L* color parameter decreased during the entire curing for all treatments. An inverse relationship between the rate of TPC decrease and enzymatic activity loss was found, but the relationship with L* was unclear. HHP appears to be an alternative vanilla killing method; nevertheless, more studies are needed to establish its clear advantages over blanching.

A novel magneto-mediated electrochemical biosensor integrated DNAzyme motor and hollow nanobox-like Pt@Ni-Co electrocatalyst as dual signal amplifiers for vanilla detection.

Herein, a novel magneto-mediated electrochemical aptasensor using the signal amplification technologies of DNAzyme motor and electrocatalyst for vanilla (VAN) detection was fabricated. The D/B duplex, formed by the DNAzyme motor that was each silenced by a blocker, and hairpin DNA1 (H1) containing adenosine ribonucleotide (rA) site were tethered on the sites of the gold nanoparticles@hollow porphyrinic-Metal-organic framework/polyethyleneimine-reduced graphene oxide (AuHPCN-222/PEI-rGO)-modified gold electrode (AuE). Then, after homogeneous and specific recognition in the presence of the VAN, trigger DNA was released and enriched by magnetic separation technique and introduced to the sensing platform to activate the DNAzyme motor, which efficiently improved target recognition capability and avoided the obstacle of multiple DNA strands tangling. More interestingly, the activated DNAzyme motor could repeatedly bind to and cleave H1 in the presence of Mg2+, leading to the exposure of a plethora of capture probes. The thionine (Thi) functionalized hairpin DNA2 (H2)-Pt@Ni-Co as signal probes could hybridize with capture probes. Additionally, the Pt@Ni-Co electrocatalysts presented catalytic activity towards Thi to obtain stronger electrochemical signals. VAN with concentrations ranging from 1 × 10-6 to 10 µM was determined and a detection limit was down to 0.15 pM. The designed electrochemical sensor was highly selective with specificity, stability, reproducibility, and reliable capability for monitoring the VAN in real samples.

Non-species-specific pollen transfer and double-reward production in euglossine-pollinated Vanilla.

Commonly attributed to orchids, the pollen movement in Vanilla has been associated with food deception and specific plant-pollinator relationships. This study investigated the role of flower rewards and pollinator specificity in the pollen transfer of a widely distributed member to the euglossinophilous Vanilla clade, V. pompona Schiede using data collected from Brazilian populations. These included investigations on morphology, light microscopy and histochemistry, and analysis of flowers scent using GC-MS. The pollinators and the mechanisms of pollination were recorded through focal observations. The yellow flowers of V. pompona are fragrant and offer nectar as reward. The major volatile compound of the V. pompona scent, carvone oxide, shows convergent evolution in Eulaema-pollinated Angiosperms. The pollination system of V. pompona is not species-specific, but its flowers are strongly adapted to pollination by large Eulaema males. Pollination mechanism is based in a combination of perfume collection and nectar seeking. The dogma of a species-specific pollination system based on food deception in Vanilla has been broken with the increase in studies on this Pantropical orchid genus. Here, least three bee species and dual reward-offering are involved in pollen transfer in V. pompona. Visitation frequency of bees collecting perfumes, used in courtship by male euglossines, is higher than in searching for food, as short-lived young euglossine males seem to be more interested in sex than food. A pollination system based on offering both nectar and perfumes as resources is described for the first time in orchids.

Recovery of flavor compounds from vanilla bagasse by hydrolysis and their identification through UPLC-MSE.

Vanilla is a globally treasured commodity, and the consequences of its unstable value affect social, environmental, economic, and academic ambits. The extensive range of aroma molecules found in cured vanilla beans is crucial to the complexity of this natural condiment and knowledge about their recovery is of the essence. Many strategies aim on reproducing the chemical intricacies of vanilla flavor, such as biotransformation and de novo biosynthesis. Few studies, however, aim at the exhaustion of the cured pods, of which the bagasse, after the traditional ethanolic extraction, might still bear a highly valued flavor composition. An untargeted liquid chromatography coupled with mass spectrometry (LC-MSE) approach was applied to elucidate if sequential alkaline-acidic hydrolysis was effective in extracting flavor related molecules and chemical classes from the hydro-ethanolic fraction. Important vanilla flavor related compounds present in the hydro-ethanolic fraction were further extracted from the residue through alkaline hydrolysis, such as vanillin, vanillic acid, 3-methoxybenzaldehyde, 4-vinylphenol, heptanoic acid, and protocatechuic acid. Acid hydrolysis was effective on further extracting features from classes such as phenols, prenol lipids, and organooxygen compounds, though representative molecules remain unknown. Finally, sequential alkaline-acidic hydrolysis rendered natural vanilla's ethanolic extraction residues as an interesting supplier of its own products, which could be used as a food additive, and many other applications.

Genome-wide identification of the glutamate receptor-like gene family in Vanilla planifolia and their response to Fusarium oxysporum infection.

Glutamate receptor-like genes (GLRs) are essential for plant growth and development and for coping with environmental (biological and non-biological) stresses. In this study, 13 GLR members were identified in the Vanilla planifolia genome and attributed to two subgroups (Clade I and Clade III) based on their physical relationships. Cis-acting element analysis and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotations indicated the GLR gene regulation's complexity and their functional diversity. Expression analysis revealed a relatively higher and more general expression pattern of Clade III members compared to the Clade I subgroup in tissues. Most GLRs showed significant differences in expression during Fusarium oxysporum infection. This suggested that GLRs play a critical role in the response of V. planifolia to pathogenic infection. These results provide helpful information for further functional research and crop improvement of VpGLRs.

Novel green production of natural-like vanilla extract from curcuminoids.

The demand for natural vanilla extract, and vanillin in particular, by far exceeds the current production, as both the cultivation of vanilla beans and the extraction of vanillin are laborious. For this purpose, most vanillin used today is produced synthetically, contrary to the general trend toward bio-based products. The present study deals with the synthesis of nature-based vanillin, starting with the more accessible rhizomes of the plant Curcuma longa. Besides vanillin, vanillic acid and p-hydroxybenzaldehyde are synthesized that way, which are also found in the natural vanilla bean. The extraction of the curcuminoids and, finally, their conversion to the flavors are performed using visible light and food-grade chemicals only. A binary mixture of ethanol and triacetin, as well as a surfactant-free microemulsion consisting of water, ethanol, and triacetin, are investigated in this context. The results exceed the literature values for Soxhlet extraction of vanilla beans by a factor > 7.

First Vanilla planifolia High-Density Genetic Linkage Map Provides Quantitative Trait Loci for Resistance to Fusarium oxysporum.

Fusarium oxysporum f. sp. radicis-vanillae (Forv), the causal agent of root and stem rot disease, is the main pathogen affecting vanilla production. Sources of resistance have been reported in Vanilla planifolia G. Jackson ex Andrews, the main cultivated vanilla species. In this study, we developed the first high-density genetic map in this species with 1,804 genotyping-by-sequencing (GBS)-generated single nucleotide polymorphism (SNP) markers using 125 selfed progenies of the CR0040 traditional vanilla cultivar. Sixteen linkage groups (LG) were successfully constructed, with a mean of 113 SNPs and an average length of 207 cM per LG. The map had a high density with an average of 5.45 SNP every 10 cM and an average distance of 1.85 cM between adjacent markers. The first three LG were aligned against the first assembled chromosome of CR0040, and the other 13 LG were correctly associated with the other 13 assembled chromosomes. The population was challenged with the highly pathogenic Forv strain Fo072 using the root-dip inoculation method. Five traits were mapped, and 20 QTLs were associated with resistance to Fo072. Among the genes retrieved in the CR0040 physical regions associated with QTLs, genes potentially involved in biotic resistance mechanisms, coding for kinases, E3 ubiquitin ligases, pentatricopeptide repeat-containing proteins, and one leucine-rich repeat receptor underlying the qFo72_08.1 QTL have been highlighted. This study should provide useful resources for marker-assisted selection in V. planifolia.

Quantitative characterizations of mOR-EG activated by vanilla odorants using advanced statistical physics modeling.

An advanced monolayer adsorption model of an ideal gas was successfully employed to investigate the adsorption of vanillin, vanillin methyl ether, vanillin ethyl ether, and vanillin acetate odorants on mouse eugenol olfactory receptor mOR-EG. In order to understand the adsorption process putatively introduced in olfactory perception, model parameters were analyzed. Hence, fitting results showed that the studied vanilla odorants were linked in mOR-EG binding pockets with a non-parallel orientation, and their adsorption was a multi-molecular process (n > 1). The adsorption energy values that ranged from 14.021 to 19.193 kJ/mol suggested that the four vanilla odorants were physisorbed on mOR-EG (ΔEa < 40 kJ/mol) and the adsorption mechanism may be considered as an exothermic mechanism (ΔEa > 0). The estimated parameters may also be utilized for the quantitative characterization of the interactions of the studied odorants with mOR-EG to determine the corresponding olfactory bands ranging from 8 to 24.5 kJ/mol.

Free comment as a valuable approach to characterize and identify the drivers of liking of high-protein flavored milk drink submitted to ohmic heating.

Flavored milk drink is a popular dairy product traditionally processed by pasteurization, which is a safe and robust process. Still, it can imply a greater energy expenditure and a more significant sensorial alteration. Ohmic heating (OH) has been proposed as an alternative to dairy processing, including flavored milk drink. However, its impact on sensory characteristics needs to be evidenced. This study used Free Comment, an underexplored methodology in sensory studies, to characterize five samples of high-protein vanilla-flavored milk drink: PAST (conventional pasteurization 72 °C/15 s); OH6 (ohmic heating at 5.22 V/cm); OH8 (ohmic heating at 6.96 V/cm); OH10 (ohmic heating at 8.70 V/cm), and OH12 (ohmic heating at 10.43 V/cm). Free Comment raised similar descriptors to those found in studies that used more consolidated descriptive methods. The employed statistical approach allowed observation that pasteurization and OH treatment have different effects on the sensory profile of products, and the electrical field strength of OH also has a significant impact. PAST was slightly to moderately negatively associated with "acid taste," "fresh milk taste," "smoothness," "sweet taste," "vanilla flavor," "vanilla aroma," "viscous," and "white color." On the other hand, OH processing with more intense electric fields (OH10 and OH12) produced flavored milk drinks strongly associated with the "in natura" milk descriptors ("fresh milk aroma" and "fresh milk taste"). Furthermore, the products were characterized by the descriptors "homogeneous," "sweet aroma," "sweet taste," "vanilla aroma," "white color," "vanilla taste," and "smoothness." In parallel, less intense electric fields (OH6 and OH8) produced samples more associated with a bitter taste, viscosity, and lumps presence. Sweet taste and fresh milk taste were the drivers of liking. In conclusion, OH with more intense electric fields (OH10 and OH12) was promising in flavored milk drink processing. Furthermore, the free comment was a valuable approach to characterize and identify the drivers of liking of high-protein flavored milk drink submitted to OH.

Morphological and Physio-Chemical Responses to PEG-Induced Water Stress in Vanilla planifolia and V. pompona Hybrids.

Vanilla planifolia is an orchid of cultural and economic value. However, its cultivation in many tropical countries is threatened by water stress. In contrast, V. pompona is a species that is tolerant of prolonged periods of drought. Due to the need for plants' resistant to water stress, the use of hybrids of these two species is considered. Therefore, the objective of this study was to evaluate the morphological and physio-chemical responses of in vitro vanilla seedlings of the parental genotype V. planifolia, and the hybrids V. planifolia × V. pompona and V. pompona × V. planifolia, which were then exposed over five weeks to polyethylene glycol-induced water stress (-0.49 mPa). Stem and root length, relative growth rate, number of leaves and roots, stomatal conductance, specific leaf area, and leaf water content were determined. Metabolites potentially associated with the response to water stress were identified in leaves, through untargeted and targeted metabolomics. Both hybrids exhibited a smaller decrease in the morphophysiological responses compared to V. planifolia and exhibited an enrichment of metabolites such as carbohydrates, amino acids, purines, phenols, and organic acids. Hybrids of these two species are considered as a potential alternative to the traditional cultivation of vanilla to face drought in a global warming scenario.

Evolutionary Patterns of the Chloroplast Genome in Vanilloid Orchids (Vanilloideae, Orchidaceae).

The Vanilloideae (vanilloids) is one of five subfamilies of Orchidaceae and is composed of fourteen genera and approximately 245 species. In this study, the six new chloroplast genomes (plastomes) of vanilloids (two Lecanorchis, two Pogonia, and two Vanilla species) were decoded, and then the evolutionary patterns of plastomes were compared to all available vanilloid plastomes. Pogonia japonica has the longest plastome, with 158,200 bp in genome size. In contrast, Lecanorchis japonica has the shortest plastome with 70,498 bp in genome size. The vanilloid plastomes have regular quadripartite structures, but the small single copy (SSC) region was drastically reduced. Two different tribes of Vanilloideae (Pogonieae and Vanilleae) showed different levels of SSC reductions. In addition, various gene losses were observed among the vanilloid plastomes. The photosynthetic vanilloids (Pogonia and Vanilla) showed signs of stage 1 degradation and had lost most of their ndh genes. The other three species (one Cyrotsia and two Lecanorchis), however, had stage 3 or stage 4 degradation and had lost almost all the genes in their plastomes, except for some housekeeping genes. The Vanilloideae were located between the Apostasioideae and Cypripedioideae in the maximum likelihood tree. A total of ten rearrangements were found among ten Vanilloideae plastomes when compared to the basal Apostasioideae plastomes. The four sub-regions of the single copy (SC) region shifted into an inverted repeat (IR) region, and the other four sub-regions of the IR region shifted into the SC regions. Both the synonymous (dS) and nonsynonymous (dN) substitution rates of IR in-cooperated SC sub-regions were decelerated, while the substitution rates of SC in-cooperated IR sub-regions were accelerated. A total of 20 protein-coding genes remained in mycoheterotrophic vanilloids. Almost all these protein genes show accelerated base substitution rates compared to the photosynthetic vanilloids. Two of the twenty genes in the mycoheterotrophic species faced strong "relaxed selection" pressure (p-value < 0.05).

Sex-Specific Alterations of the Lung Transcriptome at Birth in Mouse Offspring Prenatally Exposed to Vanilla-Flavored E-Cigarette Aerosols and Enhanced Susceptibility to Asthma.

Currently, approximately 8 million adult Americans use electronic cigarettes (e-cigs) daily, including women of childbearing age. It is known that more than 10% of women smoke during their pregnancy, and recent surveys show that rates of maternal vaping are similar to rates of maternal cigarette smoking. However, the effects of inhaling e-cig aerosol on the health of fetuses remain unknown. The objective of the present study was to increase our understanding of the molecular effects caused by in utero exposures to e-cig aerosols on developing mouse lungs and, later in life, on the offspring's susceptibility to developing asthma. METHODS: Pregnant mice were exposed throughout gestation to either filtered air or vanilla-flavored e-cig aerosols containing 18 mg/mL of nicotine. Male and female exposed mouse offspring were sacrificed at birth, and then the lung transcriptome was evaluated. Additionally, once sub-groups of male offspring mice reached 4 weeks of age, they were challenged with house dust mites (HDMs) for 3 weeks to assess asthmatic responses. RESULTS: The lung transcriptomic responses of the mouse offspring at birth showed that in utero vanilla-flavored e-cig aerosol exposure significantly regulated 88 genes in males (62 genes were up-regulated and 26 genes were down-regulated), and 65 genes were significantly regulated in females (17 genes were up-regulated and 48 genes were down-regulated). Gene network analyses revealed that in utero e-cig aerosol exposure affected canonical pathways associated with CD28 signaling in T helper cells, the role of NFAT in the regulation of immune responses, and phospholipase C signaling in males, whereas the dysregulated genes in the female offspring were associated with NRF2-mediated oxidative stress responses. Moreover, we found that in utero exposures to vanilla-flavored e-cig aerosol exacerbated HDM-induced asthma in 7-week-old male mouse offspring compared to respective in utero air + HDM controls. CONCLUSIONS: Overall, these data demonstrate that in utero e-cig aerosol exposure alters the developing mouse lung transcriptome at birth in a sex-specific manner and provide evidence that the inhalation of e-cig aerosols is detrimental to the respiratory health of offspring by increasing the offspring' susceptibility to developing lung diseases later in life.

Plant ecology: Vanilla lures both insects and mammals to disperse its seeds and fruits.

A new study shows that Vanilla species are unique among orchids for having evolved a multimodal mechanism of seed and fruit dispersal. Bees inadvertently transport seeds externally while collecting fragrance or nest material from dehiscent fruits. Rodents and marsupials consume and disperse aromatic fleshy fruits and the crustose seeds within.