Feasibility and Preliminary Efficacy of American Elderberry Juice for Improving Cognition and Inflammation in Patients with Mild Cognitive Impairment.

Despite data showing that nutritional interventions high in antioxidant/anti-inflammatory properties (anthocyanin-rich foods, such as blueberries/elderberries) may decrease risk of memory loss and cognitive decline, evidence for such effects in mild cognitive impairment (MCI) is limited. This study examined preliminary effects of American elderberry (Sambucus nigra subsp. canadensis) juice on cognition and inflammatory markers in patients with MCI. In a randomized, double-blind, placebo-controlled trial, patients with MCI (n = 24, Mage = 76.33 ± 6.95) received American elderberry (n = 11) or placebo (n = 13) juice (5 mL orally 3 times a day) for 6 months. At baseline, 3 months, and 6 months, patients completed tasks measuring global cognition, verbal memory, language, visuospatial cognitive flexibility/problem solving, and memory. A subsample (n = 12, 7 elderberry/5 placebo) provided blood samples to measure serum inflammatory markers. Multilevel models examined effects of the condition (elderberry/placebo), time (baseline/3 months/6 months), and condition by time interactions on cognition/inflammation outcomes. Attrition rates for elderberry (18%) and placebo (15%) conditions were fairly low. The dosage compliance (elderberry-97%; placebo-97%) and completion of cognitive (elderberry-88%; placebo-87%) and blood-based (elderberry-100%; placebo-100%) assessments was high. Elderberry (not placebo) trended (p = 0.09) towards faster visuospatial problem solving performance from baseline to 6 months. For the elderberry condition, there were significant or significantly trending decreases over time across several markers of low-grade peripheral inflammation, including vasorin, prenylcysteine oxidase 1, and complement Factor D. Only one inflammatory marker showed an increase over time (alpha-2-macroglobin). In contrast, for the placebo, several inflammatory marker levels increased across time (L-lactate dehydrogenase B chain, complement Factor D), with one showing deceased levels over time (L-lactate dehydrogenase A chain). Daily elderberry juice consumption in patients with MCI is feasible and well tolerated and may provide some benefit to visuospatial cognitive flexibility. Preliminary findings suggest elderberry juice may reduce low-grade inflammation compared to a placebo-control. These promising findings support the need for larger, more definitive prospective studies with longer follow-ups to better understand mechanisms of action and the clinical utility of elderberries for potentially mitigating cognitive decline.

Germplasm of Ozark chinquapin (Castanea ozarkensis Ashe) can be cryopreserved by dormant winter buds.

Ozark chinquapin (Castanea ozarkensis Ashe) is a forest tree, endemic to the Ozark Mountain region in Eastern United States. Its nutritious nuts were consumed by Native Americans, European settlers, livestock, and wild animals and its wood was an important rot-resistant construction material. Once a significant tree in regional forest communities, the species was nearly eradicated by a chestnut blight caused by Cryphonectria parasitca (Murill) Barr fungus. Some individuals have survived as sprouts from adventitious root buds, but they rarely reach reproductive maturity. While some in situ restoration efforts are underway, the development of a viable ex situ germplasm preservation method is critical to the conservation of this important food-bearing species. Our experiment aimed to develop a cryopreservation method for C. ozarkensis dormant winter buds subjected to eight experimental treatments before desiccation, slow cooling, and storage in liquid nitrogen vapor. The highest post cryogenic viability was 91.2 % for dormant buds pretreated with 0.3 M sucrose for 16 h followed by 0.75 M sucrose for 3 h; this treatment is suggested for cryopreservation of dormant winter buds of Ozark chinquapin germplasm.

Temperate silvopastures provide greater ecosystem services than conventional pasture systems.

Management and design affect systems' ability to deliver ecosystem services and meet sustainable intensification needs for a growing population. Soil-plant-animal health evaluations at the systems level for conventional and silvopastoral environments are lacking and challenge adoption across temperate regions. Impacts of silvopasture on soil quality, microclimate, cattle heat stress, forage quality and yield, and cattle weight gain were compared to a conventional pasture in the mid-southern US. Here, we illustrate silvopastures have greater soil organic carbon, water content, and overall quality, with lower temperatures (soil and cattle) than conventional pastures. Forage production and cattle weight gains were similar across systems; yet, conventional pasture systems would need approximately four times more land area to yield equivalent net productivity (tree, nuts, forage, and animal weight) of one ha of silvopasture. Temperate silvopastures enhanced delivery of ecosystem services by improving soil quality and promoting animal welfare without productivity losses, thus allowing sustainable production under a changing climate.

A comparative study of edible coatings and freshness paper on the quality of fresh North American pawpaw (Asimina triloba) fruits using TOPSIS-Shannon entropy analyses.

The North American pawpaw (Asimina triloba) is a tropical fruit that is known to be the largest edible fruit native to the United States. The fruit has remained uncommercialized because of the rapid changes in quality that occur after the fruit is harvested. However, only a few studies have evaluated the quality of the fruit during postharvest storage. This study aimed to assess the effect of different concentrations of chitosan and sodium alginate coatings, and freshness paper treatments on the quality characteristics of pawpaw fruits during storage and use TOPSIS-Shannon entropy analyses to determine which treatment best maintains the quality of the fruits from three cultivars. The results show that the chitosan coatings were more effective in slowing moisture loss in Sunflower fruits than in Susquehanna and 10-35 fruits over time. Similarly, the freshness paper treatment controlled moisture loss more effectively than sodium alginate coatings. The 10-35 fruits with 1% chitosan coating had very little change in skin color and physical appearance compared to all the other treatments. The TOPSIS-Shannon entropy analyses showed that the 10-35 fruits with 1% chitosan had the most stable quality over time, followed by the Susquehanna and Sunflower fruits with 2% chitosan coatings. The experimental data from different cultivars, treatments, and storage conditions, proved the shelf-life of pawpaw fruit could be extended from 5 days to 15-20 days depending on the cultivar. These findings will enable the creation of markets for pawpaw fruits and allow countries that grow them to generate revenue from this underutilized specialty crop.

Nuts and berries from agroforestry systems in temperate regions can form the foundation for a healthier human diet and improved outcomes from diet-related diseases.

Agroforestry is a specific type of agroecosystem that includes trees and shrubs with the potential to yield nutrient-rich products that contribute to human health. This paper reviews the literature on the human health benefits of tree nut and berry species commonly associated with agroforestry systems of the United States, considering their potential for preventing certain diet-related diseases. Emphasis is placed on those diseases that are most closely associated with poor outcomes from COVID-19, as they are indicators of confounding health prognoses. Results indicate that tree nuts reduce the risk of coronary heart disease, and walnuts (Juglans species) are particularly effective because of their unique fatty acid profile. Berries that are grown on shrubs have the potential to contribute to mitigation of hypertension, prevention of Type II diabetes, and reduced risk of cardiovascular disease. To optimize human health benefits, plant breeding programs can focus on the traits that enhance the naturally-occurring phytochemicals, through biofortification. Value-added processing techniques should be selected and employed to preserve the phytonutrients, so they are maintained through the point of consumption. Agroforestry systems can offer valuable human health outcomes for common diet-related diseases, in addition to providing many environmental benefits, particularly if they are purposefully designed with that goal in mind. The food system policies in the U.S. might be reoriented to prioritize these food production systems based on the health benefits.

Physical characterization of frozen fruits from eight cultivars of the North American pawpaw (Asimina triloba).

Pawpaw (Asimina triloba [L.] Dunal) is an underutilized fruit native to North America. The fruit has a short shelf life, and browns and softens rapidly after harvesting. These characteristics present a challenge to the advancement of pawpaw as an economically viable specialty crop. This study evaluated the physical characteristics of frozen fruits from eight cultivars of the pawpaw fruit to establish the processing potential of pawpaw fruits. The results show that freeze-thaw cycle may have influenced the peel thickness, peel color, and pulp color of the fruits. Fruits of the Susquehanna cultivar had the highest fruit weight and pulp weight, making them potentially the most suitable for pulp processing. The pawpaw fruits had almost neutral pH ranging between 6.07 ± 0.21 and 6.47 ± 0.11, which could contribute to the rapid browning on exposure to air since an acidic pH is important for slowing enzymatic browning. To aid pawpaw juice extraction, enzymatic treatments may be necessary to increase the juice yield from the pulp. Overleese fruits may be the best for pawpaw juice production. These findings can aid in the selection of processing equipment and guide processors in their efforts to utilize pawpaw fruits to avoid postharvest and post-processing losses.

Quantification of Vitamins, Minerals, and Amino Acids in Black Walnut (Juglans nigra).

This paper aims to quantify the micronutrients in black walnut and address its human health benefits. The metabolic profiling of 11 black walnut cultivars was accomplished using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight high-resolution mass spectrometer. Results revealed that the highest concentration of vitamin B9 was present in cultivar "Daniel" (avg. relative signal intensity 229.53 × 104 mAU). "Surprise" and "Daniel" cultivars had the highest amount of vitamin B5. However, vitamin A, D3, E, and K showed no significant difference among the cultivars. The vitamin content levels among the cultivars were compared by applying one way ANOVA method with (P < 0.05) significance level. Mineral analysis for the black walnut kernel, Persian walnut, and black walnut protein powder was done using Inductively Coupled Plasma Optical Emission spectroscopy. The experimental data for black walnut kernel is 0.04 mg/g for Fe and 0.03 mg/g for Zn, and for black walnut, protein powder is 0.07 mg/g for Fe and 0.07 mg/g for Zn. The amino acid analysis and comparison with black walnut kernel show that black walnut flour and protein powder have a higher amount of essential and non-essential amino acids. Therefore, researchers, food process engineers, and food product developers should consider the health benefits of black walnuts and explore the commercial potential of this native agroforestry crop.

Spatial monitoring technologies for coupling the soil plant water animal nexus.

Systems-level studies aimed at determining how soil properties are linked to plant production and ultimately animal response spatially are lacking. This study aims to identify if grazing pressure is linked to soil properties, terrain attributes, and above-ground plant accumulation and nutritive value in a silvopastoral (or integrated tree-livestock) system. Overall, cattle prefer grazing native grasses (2.81 vs. 1.24 h ha-1 AU-1) and udic (dry) landscape positions compared to aquic (wet) areas (2.07 vs. 1.60 h ha-1 AU-1). Greater grazing frequency occurs in udic soils with greater phosphorus and potassium contents and with accumulated forage with less lignin (P ≤ 0.05), which correspond to reduced elevation and greater tree height and diameter (shade) during summer mob grazing. Combining spatial monitoring technologies (both soil and animal) with forage allowance can optimize grazing systems management and sustainability spatially and temporally.

Changes in the Harpagide, Harpagoside, and Verbascoside Content of Field Grown Scrophularia lanceolata and Scrophularia marilandica in Response to Season and Shade.

Scrophularia lanceolata Pursh and Scrophularia marilandica L. are two common species within the Scrophulariaceae family that are endemic to North America. Historically, these species were used by indigenous peoples and colonialists to treat sunburn, sunstroke, frostbite, edema, as well as for blood purification, and in women's health. Several iridoid and phenylethanoid/phenylpropanoid glycosides detected in these species, such as harpagoside and verbascoside, possess anti-inflammatory and anti-nociceptive properties. Due to the presence of anti-inflammatory metabolites and the historical uses of these species, we performed a two-year field study to determine the optimal production of these important compounds. We subjected the plants to shade treatment and analyzed differences in the metabolite composition between the two species and each of their leaves, stems, and roots at various times throughout the growing seasons. We determined that S. lanceolata plants grown in full sun produced 0.63% harpagoside per dried weight in their leaves compared to shade-grown plants (0.43%). Furthermore, S. lanceolata accumulated more harpagoside than S. marilandica (0.24%). We also found that verbascoside accumulated in the leaves of S. lanceolata and S. marilandica as the growing season progressed, while the production of this metabolite remained mostly seasonally unchanged in the roots of both species.

Cyanogenic Glycoside Analysis in American Elderberry.

Cyanogenic glycosides (CNGs) are naturally occurring plant molecules (nitrogenous plant secondary metabolites) which consist of an aglycone and a sugar moiety. Hydrogen cyanide (HCN) is released from these compounds following enzymatic hydrolysis causing potential toxicity issues. The presence of CNGs in American elderberry (AE) fruit, Sambucus nigra (subsp. canadensis), is uncertain. A sensitive, reproducible and robust LC-MS/MS method was developed and optimized for accurate identification and quantification of the intact glycoside. A complimentary picrate paper test method was modified to determine the total cyanogenic potential (TCP). TCP analysis was performed using a camera-phone and UV-Vis spectrophotometry. A method validation was conducted and the developed methods were successfully applied to the assessment of TCP and quantification of intact CNGs in different tissues of AE samples. Results showed no quantifiable trace of CNGs in commercial AE juice. Levels of CNGs found in various fruit tissues of AE cultivars studied ranged from between 0.12 and 6.38 µg/g. In pressed juice samples, the concentration range measured was 0.29-2.36 µg/mL and in seeds the levels were 0.12-2.38 µg/g. TCP was highest in the stems and green berries. Concentration levels in all tissues were generally low and at a level that poses no threat to consumers of fresh and processed AE products.

Phenolic Compounds and Metals in Some Edible Annonaceae Fruits.

The concentrations of 3,4-dihydroxybenzoic acid, caffeic acid, catechin, chlorogenic acid, epicatechin, gallic acid, p-coumaric acid, quercetin, rutin, ferulic acid, and the major metals in graviola (Annona muricata), atemoya (A. squamosa x A. cherimola), fruta do conde (A. squamosa), biribá (Rollinia mucosa), and the North American pawpaw (Asimina triloba) were determined by UPLC-ESI (-)-MS/MS. It enabled the identification and quantification of phenolic compounds. Catechin was only found in atemoya, at a concentration of 38.6 µg/g-dw. Only 3,4-dihydroxybenzoic acid was found in the fruit pulps of all five fruits analyzed. Atemoya stands out for not only having catechin but also for having much more epicatechin (239 µg/g-dw) than the other fruits. At the same time, graviola had more p-coumaric acid (62.6 µg/g-dw), and the North American pawpaw had more chlorogenic acid (48.1 µg/g-dw) than the other fruits. Metals were determined by ICP equipped with axially viewed plasma. All five fruit pulps had relatively high levels of potassium, with concentrations ranging from 7640 to 15,000 µg/g-dw, with pawpaw being the lowest and atemoya being the highest. The concentrations of other metals ranged from Ca 547 to 1110, Na 14.3 to 123, P 1210 to 1690, Mg 472 to 980, Mn 1.86 to 5.27, and Zn 5.55 to 7.32 µg/g-dw. All five fruits in the Annonaceae family that were analyzed in this study have several phenolic compounds in them and were good sources of potassium, calcium, phosphorus, and magnesium.

Comparative Proteomic Analysis Unveils Critical Pathways Underlying the Role of Nitrogen Fertilizer Treatment in American Elderberry.

American elderberry (Sambucus nigra subsp. canadensis) is a rapidly growing specialty crop in Missouri and eastern North America. Nitrogen (N) is a major nutrient involved in plant growth and development. However, proteome changes for different genotypes of elder in response to varying levels of N-treatment remain undefined. To reveal plant responses to N, comparative proteomic analyses were performed to determine consistent changes in three genotypes of elderberry leaves (Adams II, Bob Gordon and Wyldewood) grown under different N-fertilizer treatments. 165 proteins separated by two dimensional gel electrophoresis showed significant differences in abundance (p < 0.05 and greater than 2-fold). Principal component analysis of the abundance profiles of these proteins revealed Bob Gordon as a distinct genotype. The 165 proteins were identified by mass spectrometry and showed similar functional distributions in these genotypes underlying the N-treatment. Among the proteins identified, 23 are mainly involved in photosynthesis, protein metabolism and redox homeostasis. Their abundance profiles were not altered upon exposure to N or genotype. These results provide novel insights into plant responses to fertilizer treatment at the proteome level and could lead to a better understanding of molecular mechanisms of elderberry growth.

Metabolomics of Two Pecan Varieties Provides Insights into Scab Resistance.

UHPLC-MS-based non-targeted metabolomics was used to investigate the biochemical basis of pecan scab resistance. Two contrasting pecan varieties, Kanza (scab-resistant) and Pawnee (scab-susceptible), were profiled and the metabolomics data analyzed using multivariate statistics. Significant qualitative and quantitative metabolic differences were observed between the two varieties. Both varieties were found to have some unique metabolites. Metabolites that were only present or more abundant in Kanza relative to Pawnee could potentially contribute to the scab resistance in Kanza. Some of these metabolites were putatively identified as quercetin derivatives using tandem mass spectrometry. This suggests that quercetin derivatives could be important to pecan scab resistance.

Discriminant Analyses of the Polyphenol Content of American Elderberry Juice from Multiple Environments Provide Genotype Fingerprint.

The cultivation of American elderberry (Sambucus nigra subsp. canadensis) continues to increase as the use of this botanical has expanded. Elderberries contain a variety of polyphenols, including anthocyanins, which have purported health benefits. However, information is lacking regarding the impact of environmental, management, and genotypic factors on the quantity and type of polyphenols and anthocyanins produced. Quantification of 16 polyphenols including eight anthocyanins present in juice from three genotypes of American elderberry grown at two Missouri sites from 2013 to 2014 was performed. Large variances in anthocyanin and other polyphenol content were observed between the different harvest seasons, locations, and genotypes. Although specific phytochemical trends due to those factors were not apparent, a discriminant analysis was able to correctly identify 45 of 48 juice samples by genotype, based on their polyphenol profiles. This type of characterization could be beneficial in elderberry authentication studies and to help develop and document high-quality dietary supplement products with specific phytochemical contents.

Purification and Characterization of a Black Walnut (Juglans nigra) Allergen, Jug n 4.

Tree nuts as a group cause a significant number of fatal anaphylactic reactions to foods. Walnuts (Juglans spp.) are one of the leading causes of allergic reactions to tree nuts in the U.S. and Japan. The purpose of this study was to purify and characterize potential food allergens from black walnut. Here, we report the isolation of the black walnuts allergen Jug n 4 (an 11S globulin) by ammonium sulfate precipitation, hydrophobic interaction, and size exclusion chromatography. Reducing SDS-PAGE analysis indicated that purified Jug n 4 consists of three major bands. N-Terminal sequencing data of these bands indicated that they were the results of a post-transcriptional protease cleavage of the mature protein at a site that consists of a known conserved protease recognition motif, NGXEET. Western blot experiments revealed that 32% of the sera from 25 patients with double-blind, placebo-controlled clinical walnut allergy contained IgE antibodies that recognized Jug n 4, indicating that it is a walnut allergen. Identifying this and additional allergens may facilitate the understanding of the allergenicity of seed storage proteins in tree nuts and their cross-reactivity.

Impact of Frozen Storage on the Anthocyanin and Polyphenol Contents of American Elderberry Fruit Juice.

The effects of frozen storage on the anthocyanin and polyphenol content of elderberry fruit juice are investigated. Juice from three genotypes of American elderberry (Adams II, Bob Gordon, and Wyldewood) was screened for total phenolic (TP) and total monomeric anthocyanin (TMA) contents with spectrophotometric methods. The individual anthocyanin content (IAC) of the juice was tested by coupling solid phase extraction with ultraperformance liquid chromatography-tandem mass spectrometry. Juice samples were tested initially upon harvest and then again after 3, 6, and 9 months of frozen storage. Juice from the three different genotypes had significantly different TP, TMA, and IAC profiles initially (p < 0.05). The TP, TMA, and IAC contents of the juice from different genotypes were significantly affected (p < 0.05) by the frozen storage time, suggesting that both genotype and length of frozen storage time can affect the anthocyanin content of elderberry fruit juice.

Inhibition of microglial activation by elderberry extracts and its phenolic components.

AIMS: Elderberry (Sambucus spp.) is one of the oldest medicinal plants noted for its cardiovascular, anti-inflammatory, and immune-stimulatory properties. In this study, we investigated the anti-inflammatory and anti-oxidant effects of the American elderberry (Sambucus nigra subsp. canadensis) pomace as well as some of the anthocyanins (cyanidin chloride and cyanidin 3-O-glucoside) and flavonols (quercetin and rutin) in bv-2 mouse microglial cells. MAIN METHODS: The bv-2 cells were pretreated with elderberry pomace (extracted with ethanol or ethyl acetate) or its anthocyanins and flavonols and stimulated by either lipopolysaccharide (LPS) or interferon-γ (IFNγ). Reactive oxygen species (ROS) and nitric oxide (NO) production (indicating oxidative stress and inflammatory response) were measured using the ROS detection reagent DCF-DA and the Griess reaction, respectively. KEY FINDINGS: Analysis of total monomeric anthocyanin (as cyanidin 3-O-glucoside equivalents) indicated five-fold higher amount in the freeze-dried ethanol extract as compared to that of the oven-dried extract; anthocyanin was not detected in the ethyl acetate extracts. Elderberry ethanol extracts (freeze-dried or oven-dried) showed higher anti-oxidant activities and better ability to inhibit LPS or IFNγ-induced NO production as compared with the ethyl acetate extracts. The phenolic compounds strongly inhibited LPS or IFNγ-induced ROS production, but except for quercetin, they were relatively poor in inhibiting NO production. SIGNIFICANCE: These results demonstrated differences in anti-oxidative and anti-inflammatory effects of elderberry extracts depending on solvents used. Results further identified quercetin as the most active component in suppressing oxidative stress and inflammatory responses on microglial cells.

Determination of Neurotoxic Acetogenins in Pawpaw (Asimina triloba) Fruit by LC-HRMS.

The concentrations of the neurotoxins, annonacin and squamocin, were determined in a lyophilized sample of the fruit pulp of the North American pawpaw (Asimina triloba) by LC coupled to high resolution mass spectrometry or LC-HRMS. The sample was extracted using dry methanol at 100 °C and 10 MPa pressure in a sealed container. The extraction of annonacin and squamocin was optimal at 100 °C with 7.72 and 0.162 mg/g, respectively, being found. Also, several isomers of annonacin and squamocin were separated and detected but not quantified.

Peptidomics study of anthocyanin-rich juice of elderberry.

Biologically active peptides play a role in plant signaling and defense. Elderberry juice is known to contain a variety of anthocyanin compounds, a sub-set of polyphenols, which are responsible for the deep purple color of the juice. In this paper, we describe a method utilizing solid phase extraction (SPE) to remove anthocyanins from peptides. Liquid chromatography coupled with tandem mass spectrometry was used to separate and identify the peptides. The results showed that the use of SPE was an effective method to separate peptides from anthocyanins and other background compounds including high polyphenol content in the juice samples. More than 1000 peptides present in elderberry juice were successfully identified.