Chemical Constituents and Antioxidant Potential of Red Guava (Psidium cattleianum Sabine) from Southern Brazil in Different Edible Ripening Stages.

Ripening and growing location are important factors that can impact fruit quality characteristics. In this study, the influence of these factors on physicochemical characteristics, carbohydrates, aliphatic organic acids, phenolic compounds, and antioxidant capacity of red guava (Psidium cattleianum Sabine) was evaluated. Fruit ripening increased fructose and glucose (up to 22.83 and 16.42 g 100 g- 1 dry matter (DM), respectively), and decreased citric acid, the major organic acid (up to 135.35 mg g- 1 DM). Ripening and growing location also influenced the concentration of phenolic compounds and antioxidant capacity of red guava, in which a dependency between both factors was observed in most cases. Apigenin, galangin, isoquercitrin, among other phenolic compounds were quantified for the first time in red guava, in which isoquercitrin was the major (up to 13409.81 mg kg- 1 DM). The antioxidant potential of red guava was also confirmed by ferric reducing antioxidant power assay (up to 82.63 µmol Fe+ 2 g- 1 DM), Folin-Ciocalteu reducing capacity assay (up to 17.79 mg gallic acid equivalent g- 1 DM), and DPPH free radical scavenging assay (up to 25.36 mg ascorbic acid equivalent g- 1 DM). These results especially demonstrated the bioactive potential of red guava and provided knowledge regarding the influence of ripening and growing location on chemical and bioactive components encouraging its industrial exploitation.

Free amino acid profile in Mimosa scabrella honeydew honey from Brazil and chemometric analysis for geographical discrimination.

In this study, twenty free amino acids (FAA) were investigated in samples of bracatinga (Mimosa scabrella) honeydew honey (BHH) from Santa Catarina (n = 15) and Paraná (n = 13) states (Brazil), followed by chemometric analysis for geographic discrimination. The FAA determination was performed by gas chromatography-mass spectrometry (GC-MS) after using a commercial EZ:faast™ kits for GC. Eight FAA were determined, being proline, asparagine, aspartic and glutamic acids found in all BHH, with significant differences (p < 0.05). In addition, with the exception of proline, the others FAA (asparagine, aspartic and glutamic) normally showed higher concentrations in samples from Santa Catarina state, being that in these samples it was also observed higher FAA sums (963.41 to 2034.73 mg kg-1) when compared to samples from Paraná state. The variability in the results did not show a clear profile of similarity when the heatmap and hierarchical grouping were correlated with the geographic origin and the concentration of eight determined FAA. However, principal component analysis (PCA) demonstrated that serine, asparagine, glutamic acid, and tryptophan were responsible for the geographic discrimination among samples from Santa Catarina and Paraná states, since they were the dominant variables (r > 0.72) in the PCA. Therefore, these results could be useful for the characterization and authentication of BHH based on their FAA composition and geographic origin.

Kinetic Profile of Urine Metabolites after Acute Intake of a Phenolic Compounds-Rich Juice of Juçara (Euterpe edulis Mart.) and Antioxidant Capacity in Serum and Erythrocytes: A Human Study.

The juçara palm tree produces a small spherical and black-purple fruit similar to açaí. It is rich in phenolic compounds, especially anthocyanins. A clinical trial evaluated the absorption and excretion of the main bioactive compounds in urine and the antioxidant capacity in serum and erythrocytes of 10 healthy subjects after juçara juice intake. Blood samples were collected before (0.0 h) and 0.5 h, 1 h, 2 h, and 4 h after a single dose (400 mL) of juçara juice, while urine was collected at baseline and 0-3 and 3-6 h after juice intake. Seven phenolic acids and conjugated phenolic acids were identified in urine deriving from the degradation of anthocyanins: protocatechuic acid, vanillic acid, vanillic acid glucuronide, hippuric acid, hydroxybenzoic acid, hydroxyphenylacetic acid, and ferulic acid derivative. In addition, kaempferol glucuronide was also found in urine as a metabolite of the parent compound in juçara juice. Juçara juice caused a decrease in the total oxidant status of serum after 0.5 h in comparison to baseline values (p < 0.05) and increased the phenolic acid metabolites excretion. This study shows the relationship between the production of metabolites of juçara juice and the total antioxidant status in human serum, indicating evidence of its antioxidant capacity.

Brazilian stingless bee honey: Physicochemical properties and aliphatic organic acids content.

This study aimed to investigate for the first time fourteen aliphatic organic acids (AOA) in honeys produced by different species of Brazilian stingless bees (Melipona bicolor, Scaptotrigona bipunctata, Melipona quadrifasciata, and Melipona marginata) and characterize them regarding their physicochemical properties. Thirteen AOAwere quantified in the samples, in which five of them (malonic, fumaric, glycolic, glutaric, and propionic acids) were identified for the first time instingless bee honey (SBH). Acetic, gluconic, and lactic acids were predominant in all the samples analyzed varying from 0.0067 ± 0.0001 to 1.5993 ± 0.0003 g 100 g-1, 0.0808 ± 0.0007 to 1.3460 ± 0.0006 g 100 g-1, and 0.0370 ± 0.000 to 0.5760 ± 0.0006 g 100 g-1, respectively. Most physicochemical properties showed significant differences (p < 0.05) among the samples. However, it was observed that the water activity (Aw) did not differ significantly between honey samples produced by the same species. Moreover, it is important to highlight the high moisture content, Aw, and free acidity that were found in the range of 29.6 to 40.1 g 100 g-1, 0.75 to 0.84, and 37.8 to 123 mEq kg-1, respectively. This information reinforces such peculiar characteristics of SBH and a need to deeply investigate the physical and chemical characteristics of honey from different species of stingless bees. In conclusion, it was observed that the honey samples of the different stingless bee species presented a great variation regarding their AOA content, highlighting acetic, gluconic, and lactic acids as the major AOA in all the samples. However, since this was an exploratory study, it was not possible to find any correlation between honey produced by the same species.

Grumixama (Eugenia brasiliensis Lamarck) functional phytochemicals: Effect of environmental conditions and ripening process.

Grumixama (Eugenia brasiliensis Lamarck) is a native fruit of the Atlantic rain forest that belongs to Myrtaceae family. It presents economic potential due to the attractive sensory attributes and bioactive compounds. This study determined physicochemical characteristics, minerals (K, Na, Ca, Mg, Mn), sugars (fructose, glucose, sucrose), total content of phenolics, proanthocyanidins, and anthocyanins, individual phenolic compounds, and antioxidant capacity at three ripening stages of grumixama from two growing locations (Florianópolis and São Ludgero, Santa Catarina state, Brazil). Of the 23 phenolic compounds quantified, gallic acid, ellagic acid, quercetin, and isoquercetrin were the major (with values up to 69.08 mg/100 g fresh weight). Fructose (up to 5.28 g/100 g fresh weight) was the main sugar, and K (up to 589.30 mg/100 g fresh weight) was the major mineral, followed by Ca, Na, and Mg (up to 149.02, 117.52, and 80.18 mg/100 g fresh weight, respectively). More mature fruits had the highest levels of total phenolics (up to 469.18 mg gallic acid equivalent/100 g fresh weight), anthocyanins (up to 518.64 mg cyanidin 3-glucoside equivalent/100 g fresh weight), and proanthocyanidins (up to 420.70 mg catechin equivalent/100 g fresh weight). The influence of growing location and ripening on nutritional and physicochemical characteristics was confirmed. There was a statistically significant interaction (p < 0.05) between both factors for almost all parameters evaluated. The results showed that grumixama fruits could contribute to the daily intake of nutritional and bioactive compounds for consumers' health.

Phenolic profile and in vitro anti-inflammatory activity of Mimosa scabrella Bentham honeydew honey in RAW 264.7 murine macrophages.

The anti-inflammatory activity is mainly attributed to the phenolic compounds. Once the geographical location affects the phenolic content of honeys, a relationship between the collection spot and the anti-inflammatory effect of bracatinga (Mimosa scabrella Bentham) honeydew honeys was hypothesized. The inhibitory effect of 14 honey samples on NOx, TNF-α, IL-6, IL-12p70, MCP-1, INF-γ, and IL-10 in RAW 264.7 macrophages inflamed by LPS was evaluated. Fourteen phenolic compounds were identified, mainly syringic acid and rutin. Ten honeys inhibited nitrite production; at least six downregulated TNF-α, IL-12p70, MCP-1, and IFN-γ; only four honey samples inhibited IL-6; and one honey sample inhibited IL-10 levels, showing their variable effects on the inflammatory markers. Principal component analysis grouped samples according to the phenolic content and downregulation of specific inflammatory markers. The bracatinga honeydew honey effectiveness was associated with geographical location, as samples from areas with higher density and diversity of plants had a more significant anti-inflammatory effect. PRACTICAL APPLICATIONS: The present research study investigated the anti-inflammatory potential of bracatinga honeydew honey samples collected from regions with different vegetation coverages. Honey samples collected from locations presenting greater forest diversity and density inhibited inflammatory markers more efficiently. This study reinforces the role of the bracatinga honeydew honey in preventing inflammatory processes and the importance of preserving forests so that products with a greater diversity of compounds and consequently more active can be obtained.

Assessment of Sorbate and Benzoate Content in Mustard, Ketchup and Tomato Sauce by Sub-Minute Capillary Electrophoresis.

RESEARCH BACKGROUND: Sorbate and benzoate are important preservatives in food products, but these compounds can also have genotoxic effects, causing health risks to consumers. In this regard, this study aims to determine the mass fractions of sorbate and benzoate in Brazilian samples of mustard, ketchup and tomato sauce using an adequately validated sub-minute capillary electrophoresis method. EXPERIMENTAL APPROACH: In this study, sorbate and benzoate were evaluated in sauce samples by capillary electrophoresis using a simple sample preparation procedure. Previously, the method was validated according to Eurachem guidelines, and its greenness was assessed by Eco-Scale. RESULTS AND CONCLUSIONS: The fitness for purpose of the method, as well as its suitability for the analysis of the studied matrices and its agreement with the principles of green chemistry were checked and confirmed. Also, according to our findings, among the 30 commercial samples assessed, six of them presented some mislabeling or non-compliance with European or Brazilian legislation, reinforcing the constant need for quality assessment and surveillance of food products. NOVELTY AND SCIENTIFIC CONTRIBUTION: So far, there have been few studies related to investigating the preservatives such as sorbate and benzoate in mustard, ketchup and tomato sauce, highlighting the significance and contribution of the obtained results to the knowledge in the field.

Effect of long-term and heating storage on honey visible spectrum: an alternative parameter for quality monitoring of bracatinga honeydew honey.

Bracatinga (Mimosa scabrella Bentham) honeydew honey (BHH) is a peculiar Brazilian honey. It is produced only every 2 years, which raises concerns about its quality since it can be submitted to different storage conditions until a new harvest is carried out. Therefore, this study investigated the changes in the visible spectrophotometric profile (VSP) of BHH during its storage at room temperature over 24 months and 40 °C for 4 months. Our findings indicated a similar VSP between the BHH samples, but that varied according to the storage condition. These changes were associated with the formation of brown compounds, such as 5-hydroxymethylfurfural, which has a maximum limit established for honeys. Thereby, absorbance above 0.500 absorption units between 380 and 410 nm was proposed as indicative of BHH exposure to prolonged heating with significant loss of its quality. Still, a regression model for absorbance at 380 nm was proposed aiming to predict the BHH storage time at room temperature, since storage time longer than 20 months at average temperatures of 23.0 ± 2.3 °C do not seem to be suitable for BHH. Thus, the VSP showed potential for monitoring BHH quality.

Phenolic composition and biological activities of stingless bee honey: An overview based on its aglycone and glycoside compounds.

Stingless bees are native to tropical and subtropical countries, such as Brazil. The wide variety of species, the sources of food collection (nectar and pollen), and the climate conditions strongly affect the chemical composition of the honey, making this a unique product with peculiar characteristics. Stingless bee honey presents higher water content, higher acidity, and a lower sugar concentration when compared to Apis mellifera honey. Moreover, there is a wide variety of microorganisms in stingless bees' environment, which leads their honey to go through a natural fermentative process during its production in the hive. Besides, fermentation and hydrolysis are effective ways to convert glycosides into aglycones, thus increasing the bioavailability of compounds. In this sense, stingless bee honey may possess a greater concentration of phenolic compounds aglycones than glycosides, which would increase its potential benefits. Therefore, this review aims to compile the most recent studies of stingless bee honey phenolic profile and its biological potential (antioxidant, antimicrobial, and anti-inflammatory activities) and a possible connection to its natural fermentation process.

Physicochemical properties and biological activities of bracatinga honeydew honey from different geographical locations.

Bracatinga (Mimosa scabrella Bentham) honeydew honey is a Brazilian dark honey in increasing international appreciation. In this sense, the knowledge of its composition and potential biological properties becomes indispensable. In the present study, the physicochemical characteristics, including mineral and phenolic composition, and the scavenging, reducing, and antimicrobial proprieties of bracatinga honeydew honey (bhh) from five different geographical locations, were investigated. Bhh proved to be a potential functional food due to its high content of minerals (up to 6395 mg kg-1) and phenolic compounds (up to 2393 µg 100 g-1) and high scavenging and reducing activities. High antimicrobial activity against four bacterial strains, with minimum inhibitory concentration values ranging from 10 to 60%, were also found. Additionally, through principal component analysis, partial discrimination of bhh was observed according to the geographical location, which favored the separation of samples from Lages, and mainly due to the presence of nectar in this honey, which was proposed for the samples from Bom Retiro. SUPPLEMENTARY INFORMATION: The online version of this article (10.1007/s13197-020-04937-x) contains supplementary material, which is available to authorized users.

Quantification of pyrrolizidine alkaloids in Senecio brasiliensis, beehive pollen, and honey by LC-MS/MS.

This article presents the determination of eight pyrrolizidine alkaloids (PAs) by LC-MS/MS in honeys, pollen, and Senecio brasiliensis (Asteraceae) samples, all from Santa Catarina state, Brazil. In addition, the Box-Behnken design was used to perform an optimized sample preparation on pollens and S. brasiliensis parts. Senecionine and its N-oxide, besides retrorsine N-oxide, were determined in six of the seven honeys samples. Pollen from species of the Asteraceae, Fabaceae, and Boraginaceae families were found with greater predominance in three of the seven honeys samples. In these three honeys samples were also found the highest PAs levels. In beehive pollen, flower, and leaf of S. brasiliensis, the total levels of PAs and their N-oxides reached 221, 14.1 × 104, and 14.8 × 104 mg kg-1, respectively. In honeys, these compounds are chemical contaminants and therefore undesirable when the sum exceeds 71 µg kg-1, according to EFSA. On the other hand, although PAs are naturally present in plant and pollen of some species (Senecio, Crotalaria, Bacharis, Ecchium, Mimosa scabrella, Vernonia), it is important to monitor their levels in plants but also in honeys, and other beehive products since these compounds are transferred to the final product.

Quality, composition and health-protective properties of citrus honey: A review.

Citrus honey is one of the most important monofloral honeys produced and consumed worldwide. This honey has pleasant sensorial characteristics, which include light color and typical aroma and flavor. Besides that, several constituents such as minerals, phenolic and volatile compounds, amino acids, sugars, enzymes, vitamins, methylglyoxal and organic acids are found in citrus honey. Moreover, potential biological properties have been associated with citrus honey. All these factors make it highly desired by consumers, increasing its market value, which can stimulates the practice of fraud. Also, citrus honey is susceptible to contamination and to inadequate processing. All these factors can compromise the quality, safety and authenticity of citrus honey. In this sense, this review aims to update and to discuss, for the first time, the data available in the literature about the physicochemical and the sensorial characteristics, composition, health properties, contamination, authenticity and adulteration of citrus honey. With this background, we aim to provide data that can guide future researches related to this honey.

Current status of the gastrointestinal digestion effects on honey: A comprehensive review.

In the past five years, more than 8000 scientific reports have been published on honey composition and its potential bioactivity as a source of pro-health components. However, the potential effectiveness of nutrients and other compounds in the human body is greatly influenced by the individual digestion conditions. Consequently, changes in the structure of honey components and their interactions with other constituents are expected and they may affect the bioaccessibility, the bioavailability, and further physiological functions of honey nutrients and bioactives. In this context, in addition to present key physiological characteristics for each step of the human digestion and their simulation aspects, this review also summarizes and discusses available data regarding the effect of the digestion (in vitro and in vivo) on honey compounds. Additionally, we consider the influence of the digestion on biological activities described for the compounds in the honey.

Comparative quantification and differentiation of bracatinga (Mimosa scabrella Bentham) honeydew honey proteins using targeted peptide markers identified by high-resolution mass spectrometry.

Honey traceability is an important topic, especially for honeydew honeys, due to the increased incidence of adulteration. This study aimed to establish specific markers to quantify proteins in honey. A proteomics strategy to identify marker peptides from bracatinga honeydew honey was therefore developed. The proteomics approach was based on initial untargeted identification of honey proteins and peptides by LC-ESI-Triple-TOF-MS/MS, which identified the major royal jelly proteins (MRJP) presence. Afterwards, the peptides were selected by the in silico digestion. The marker peptides were quantified by the developed targeted LC-QqQ-MS/MS method, which provided good linearity and specificity, besides recoveries between 92 and 100% to quantify peptides from bracatinga honeydew honey. The uniqueness and high response in mass spectrometry were backed by further complementary protein analysis (SDS-PAGE). The selected marker peptides EALPHVPIFDR (MRJP 1), ILGANVK (MRJP 2), TFVTIER (MRJP 3), QNIDVVAR (MRJP 4), FINNDYNFNEVNFR (MRJP 5) and LLQPYPDWSWTK (MRJP 7), quantified by LC-QqQ-MS/MS, highlighted that the content of QNIDVVAR from MRJP 4 could be used to differentiate bracatinga honeydew honey from floral honeys (p < 0.05) as a potential marker for its authentication. Finally, principal components analysis highlighted the QNIDVVAR content as a good descriptor of the analyzed bracatinga honeydew honey samples.

In vitro anti-inflammatory properties of honey flavonoids: A review.

Honey is a natural ready-to-eat product rich in flavonoids, which is known by the wound healing properties due to both antibacterial and antioxidant activity. Flavonoids mitigate inflammatory processes, and thus it could currently support studies of anti-inflammatory potential of honeys. In this review, in vitro anti-inflammatory properties of flavonoids found in honey were prioritized. Mechanistic information of specific isolated flavonoids as modulators of inflammatory processes are summarized aiming to stimulate studies regarding the action of honey in inflammatory events. Lastly, a structure-activity relationship (SAR) of flavonoids was also included. Flavonoids found in honey have demonstrated antioxidant properties and ability to inhibit pro-inflammatory enzymes such as COX, LOX, iNOS, and pro-inflammatory mediators, including nitric oxide, cytokines and chemokines. Transcriptional factors such as NF-κB are also modulated by flavonoids, controlling the expression of several inflammatory mediators. SAR studies demonstrate the effect of flavonoids in the prevention of inflammatory cascades. Despite the promising reports of in vitro anti-inflammatory activity, well-designed clinical trials need yet to be performed to confirm the benefits of honeys from different botanical sources in diseases that include episodes of inflammation.

Aliphatic organic acids as promising authenticity markers of bracatinga honeydew honey.

This study aimed to differentiate bracatinga (Mimosa scabrella Bentham) honeydew honey (BHH) from blossom honeys and BHH intentionally adulterated, all of them from three harvests, associating data of aliphatic organic acids (AOA) determined by capillary electrophoresis and chemometric analyses. The profile and concentration of AOA in pure BHH were similar between harvests, but distinct from blossom honeys. Succinic, glycolic, glutaric, malic, acetic, gluconic, and lactic acids were responsible for the differentiation between these two types of honey since they were the dominant variables (r > 0.80) in the principal component analysis. Based on this, the classification and regression trees method was used to develop a classification model considering these AOA. The proposed method needed only six of these AOA and adequately classified all blossom honeys and almost all pure and adulterated BHH. Therefore, the proposed model proved to be promising and reliable for verifying authenticity and fraud detection in BHH.

Pyrrolizidine alkaloids and beehive products: A review.

Pyrrolizidine alkaloids (PA) are secondary metabolites of plants, which are mostly found in the genus Senecio, Echium, Crotalaria, and Eupatorium. The presence of 1,2-unsaturated PA in foods is a concern to food regulators around the world because these compounds have been associated to acute and chronic toxicity, mainly in the liver. The intake foods with PA/PANO usually occur through accidental ingestion of plants and their derivatives, besides to products of vegetal-animal origin, such as honey. PA/PANO are transferred to honey by their presence in nectar, honeydew, and pollen, which are collected from the flora by bees. In addition to honey, other beekeeping products, such as pollen, royal jelly, propolis, and beeswax, are also vulnerable to PA contamination. In this context, this review provides information about chemical characteristics, regulation, and toxicity, as well as summarizes and critically discusses scientific publications that evaluated PA in honeys, pollens, royal jelly, and propolis.

Composition and potential health effects of dark-colored underutilized Brazilian fruits - A review.

Among the rich Brazilian biodiversity, a wide range of native and exotic fruit species are found. Many of these fruits have high nutritional and bioactive value, being important sources of minerals, phenolic compounds, vitamins, dietary fiber, among others. Also, the prevention of several diseases and disorders have being associated to the action of phytochemical compounds present in fruits with antioxidant, anti-inflammatory, and antiproliferative effects, for example. However, despite the potential of these fruits to be explored economically, many of them remain unknown to a large part of the population and food, pharmaceutical, and cosmetical industries, being produced and consumed only locally. Therefore, summarize information about potential Brazilian native fruits species is essential to stimulate their better use and reinforce the local economy promoting the cultivation and exploration of these fruits. In this context, this review summarizes and discusses the physicochemical characteristics, nutrients, phytochemical compounds, and biological properties of seven dark-colored underexploited Brazilian fruits species: Bactris setosa, Eugenia brasiliensis, Eugenia involucrata, Euterpe edulis, Myrcianthes pungens, Myrciaria cauliflora, and Myrciaria jaboticaba.

Use of visible spectrophotometric fingerprint and chemometric approaches for the differentiation of Mimosa scabrella Bentham honeydew honey.

Honeydew honeys, including bracatinga (Mimosa scabrella Bentham) honeydew honey, are highly appreciated due to its nutritional and sensory properties. Therefore, fast and inexpensive methods must be developed to differentiate this distinct class of honey. Although methods using the visible spectrophotometric fingerprint (VSF) have never been used to differentiate bracatinga honeydew honey and blossom honey, this technique appears to be a viable alternative, successfully used in the differentiation and fraud detection of various foods, including honeys from other botanical sources. In this sense, the present study aimed to verify the existence of a specific VSF for bracatinga honeydew honey and blossom honey and differentiate both type of honeys using the VSF associated with chemometric analysis. The VSF of 30 bracatinga honeydew honeys harvested in three different years (2014, 2016, and 2018) and 21 blossom honeys (harvested in 2016) were evaluated. The existence of a different VSF for bracatinga honeydew honeys and blossom honeys allowed the satisfactory differentiation of both types of honeys. Additionally, bracatinga honeydew honey presented a unique VSF, independent of the year of harvest, which can be exploited as a fingerprint of this type of honey, contributing to its authenticity.

Açaí (Euterpe oleracea Mart.) and juçara (Euterpe edulis Mart.) juices improved HDL-c levels and antioxidant defense of healthy adults in a 4-week randomized cross-over study.

OBJECTIVE: To evaluate the effects of moderate-term açaí and juçara juice intake on fasting glucose, lipid profile, and oxidative stress biomarkers in healthy subjects. METHODS: A randomized cross-over study was performed with 30 healthy adults. The subjects were assigned to drink 200 mL/day of açaí or juçara juice for four weeks with a 4-week washout period. Before and after each nutritional intervention, blood samples were obtained to evaluate the outcomes: fasting glucose, total cholesterol, triglycerides, high-density lipoprotein-cholesterol (HDL-c), low-density lipoprotein-cholesterol (LDL-c), small, dense LDL-c (sd-LDL-c), total antioxidant capacity (TAC), total oxidant status (TOS), oxidative stress index (OSI), uric acid, and activity of the enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). RESULTS: After four weeks, açaí and juçara juices increased the concentrations of HDL-c by 7.7% and 11.4%, respectively (P < 0.05). In addition, açaí juice intake promoted significant increases in TAC (66.7%), CAT (275.1%), GPx (15.3%), and a decrease in OSI (55.7%) compared to baseline (P < 0.05 for all). Juçara juice intake significantly increased CAT activity (~15.0%) in relation to baseline. No significant intergroup differences were observed for any outcomes (P > 0.05). CONCLUSION: The results indicated a positive impact of regular consumption of açaí and juçara juices on the HDL-c levels, as well as on the antioxidant enzyme activities, which may contribute to cardiovascular health.