Using urinary metabolomics to identify metabolic pathways linked to cardiac structural alterations in young adults: The African-PREDICT study.

BACKGROUND AND AIMS: Risk factor exposure from young ages was shown to contribute to cardiovascular events - cardiac hypertrophy, which may be accompanied by an altered metabolism. To determine how early metabolic alterations associate with myocardial structural changes, we profiled urinary metabolites in young adults with cardiovascular disease (CVD) risk factor(s) and a control group without CVD risk factors. METHODS AND RESULTS: We included healthy adults (N = 1202), aged 20-30 years, stratified based on risk factors, i.e., obesity, physical inactivity, elevated blood pressure (BP), hyperglycemia, dyslipidemia, low socio-economic status, smoking and excessive alcohol use - forming the CVD risk group (N = 1036) and the control group (N = 166). Relative wall thickness (RWT) and left ventricular mass index (LVMi) were measured using echocardiography. Targeted metabolomics data were obtained using a liquid chromatography-tandem mass spectrometry method. Clinic systolic BP, 24 h BP and RWT were higher in the CVD risk group compared to the control group (all P ≤ 0.031). Exclusively in the CVD risk group, RWT associated with creatine and dodecanoylcarnitine; while LVMi associated with glycine, serine, glutamine, threonine, alanine, citrulline, creatine, proline, pyroglutamic acid and glutamic acid (all P ≤ 0.040). Exclusively in the control group, LVMi associated with propionylcarnitine and butyrylcarnitine (all P ≤ 0.009). CONCLUSION: In young adults without CVD, but with CVD risk factors, LVMi and RWT associated with metabolites linked energy metabolism (shifting from solely fatty acid oxidation to glycolysis, with impaired creatine kinase activity) and oxidative stress. Our findings support early onset metabolic changes accompanying cardiac structural alterations due to lifestyle and behavioural risk factors.

Markers of arterial stiffness and urinary metabolomics in young adults with early cardiovascular risk: the African-PREDICT study.

INTRODUCTION: Increased exposure to risk factors in the young and healthy contributes to arterial changes, which may be accompanied by an altered metabolism. OBJECTIVES: To increase our understanding of early metabolic alterations and how they associate with markers of arterial stiffness, we profiled urinary metabolites in young adults with cardiovascular disease (CVD) risk factor(s) and in a control group without CVD risk factors. METHODS: We included healthy black and white women and men (N = 1202), aged 20-30 years with a detailed CVD risk factor profile, reflecting obesity, physical inactivity, smoking, excessive alcohol intake, masked hypertension, hyperglycemia, dyslipidemia and low socio-economic status, forming the CVD risk group (N = 1036) and the control group (N = 166). Markers of arterial stiffness, central systolic blood pressure (BP) and pulse wave velocity were measured. A targeted metabolomics approach was followed by measuring amino acids and acylcarnitines using a liquid chromatography-tandem mass spectrometry method. RESULTS: In the CVD risk group, central systolic BP (adjusted for age, sex, ethnicity) was negatively associated with histidine, arginine, asparagine, serine, glutamine, dimethylglycine, threonine, GABA, proline, methionine, pyroglutamic acid, aspartic acid, glutamic acid, branched chain amino acids (BCAAs) and butyrylcarnitine (all P ≤ 0.048). In the same group, pulse wave velocity (adjusted for age, sex, ethnicity, mean arterial pressure) was negatively associated with histidine, lysine, threonine, 2-aminoadipic acid, BCAAs and aromatic amino acids (AAAs) (all P ≤ 0.044). In the control group, central systolic BP was negatively associated with pyroglutamic acid, glutamic acid and dodecanoylcarnitine (all P ≤ 0.033). CONCLUSION: In a group with increased CVD risk, markers of arterial stiffness were negatively associated with metabolites related to AAA and BCAA as well as energy metabolism and oxidative stress. Our findings may suggest that metabolic adaptations may be at play in response to increased CVD risk to maintain cardiovascular integrity.

White wine phenolics: current methods of analysis.

White wine phenolic analyses are less common in the literature than analyses of red wine phenolics. Analytical techniques for white wine phenolic analyses using spectrophotometric, chromatographic, spectroscopic, and electrochemical methods are reported. The interest of research in this area combined with the advances in technology aimed at the winemaking industry are promoting the establishment of novel approaches for identifying, quantifying, and classifying phenolic compounds in white wine. This review article provides an overview of the current research into white wine phenolics through a critical discussion of the analytical methods employed. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Urinary metabolomics profiling by cardiovascular risk factors in young adults: the African Prospective study on Early Detection and Identification of Cardiovascular disease and Hypertension study.

AIM: Risk factors contributes to a dysregulated metabolism and may ultimately increase the predisposition for cardiovascular disease (CVD) development. To increase our understanding of mechanistic pathways associated with CVD risk, we profiled the urinary metabolome according to individual and clusters of CVD risk factors in comparison with a control group without any risk factors. METHODS AND RESULTS: Healthy black and white women and men ( N  = 1202), aged 20-30 years with a detailed CVD risk factor profile were included. CVD risk groups: obese, physical inactive, smoking, excessive alcohol intake, masked hypertensive, hyperglycaemic, dyslipidemic and low socioeconomic status. CVD risk clusters were based on the presence of 1, 2 and 3 or more risk factors. Liquid chromatography-tandem mass spectrometry was used to obtain urinary metabolomics data (amino acids and acylcarnities). Compared with the control group, higher levels of metabolites associated with aromatic and branched chain amino acid metabolism including phenylalanine, tyrosine and leucine/isoleucine were found in the obese, masked hypertensive, hyperglycaemic, low socioeconomic groups (all q  ≤ 0.032) and 3+ CVD risk cluster (all P  ≤ 0.034). Metabolites associated with the y-glutamyl cycle including glycine, histidine, serine, glutamine, methionine, cystine and pyroglutamic acid were found in the hyperglycaemic, low socioeconomic groups (all q  ≤ 0.050), 2 and 3+ CVD risk clusters (all P  ≤ 0.041). Metabolites associated with energetics including acetylcarnitine (lower levels), hexanoylcarnitine and decanoylcarnitine were found in the low socioeconomic group, 1 and 3+ CVD risk clusters ( q / P  ≤ 0.050) ( ß -oxidation). In addition to the above-mentioned amino acids, alanine and threonine were found in the hyperglycaemic, low socioeconomic groups, 2 and 3+ CVD risk clusters (all q / P  ≤ 0.047) (glycolysis). Creatine in the obese, hyperglycaemic groups (all q  ≤ 0.049) and 3+ CVD risk cluster (all P  ≤ 0.041) (creatine pathway). CONCLUSION: Exposure to CVD risk factors is associated with a dysregulated metabolism in the above-mentioned pathways that may precede the development of CVD.

The effect of enzyme treatment on polyphenol and cell wall polysaccharide extraction from the grape berry and subsequent sensory attributes in Cabernet Sauvignon wines.

Pectolytic enzyme maceration is common for producing red wines, but the effects on bitterness and astringency are not well understood. Glycan microarrays assessed polysaccharide diversity and with polyphenol analysis was correlated with sensory data on descriptors of astringency and their perceived levels in enzyme-crafted Cabernet Sauvignon wines. Enzyme use is shown to have no effect on bitterness, but enzyme-macerated wines are more astringent. The data suggests that pectolytic enzymes are much more pronounced in their effect on the cell wall matrix than the ripeness of the berries at harvest and subsequent sensory perception. Enzyme-macerated red wines showed higher levels of polyphenol which were more polymerized and galloylated. The polyphenol-rich wines were described as hard, chalky, grippy, grainy and dry. The non-enzyme wines had elevated levels of arabinogalactan protein and pectin epitopes (notably biomarker mAbs JIM8 and JIM13) with the wines being characterized as soft, fine and velvety.

Relationship between anthocyanins, proanthocyanidins, and cell wall polysaccharides in grapes and red wines. A current state-of-art review.

Numerous research studies have evaluated factors influencing the nature and levels of phenolics and polysaccharides in food matrices. However, in grape and wines most of these works have approach these classes of compounds individually. In recent years, the number of publications interconnecting classes have increased dramatically. The present review relates the last decade's findings on the relationship between phenolics and polysaccharides from grapes, throughout the entire winemaking process up to evaluating the impact of their relationship on the red wine sensory perception. The combination and interconnection of the most recent research studies, from single interactions in model wines to the investigation of the formation of complex macromolecules, brings the perfect story line to relate the relationship between phenolics and polysaccharides from the vineyard to the glass. Grape pectin is highly reactive toward grape and grape derived phenolics. Differences between grape cultivars or changes during grape ripeness will affect the extractability of these compounds into the wines. Therefore, the nature of the grape components will be crucial to understand the subsequent reactions occurring between phenolics and polysaccharide of the corresponding wines. It has been demonstrated that they can form very complex macromolecules which affect wine color, stability and sensory properties.

Direct quantification of red wine phenolics using fluorescence spectroscopy with chemometrics.

Phenolic compounds are secondary metabolites known to play crucial roles in important chemical reactions impacting the mouthfeel, colour and ageing potential of red wine. Their complexity has resulted in a number of advanced analytical methods, which often prevent routine phenolic analysis in winemaking. Fluorescence spectroscopy could be an alternative to current spectrophotometric techniques and its combination with chemometrics was investigated for its suitability in directly quantifying phenolic content of unaltered red wine and fermenting samples. Front-face fluorescence was optimised and used to build predictive models for total phenols, total condensed tannins, total anthocyanins, colour density and polymeric pigments. Machine learning algorithms were used for model development. The most successful models were built for total phenols, total condensed tannins and total anthocyanins with coefficient of determination (R2cal) and RMSECV of 0.81, 0.89, 0.80 and 5.71, 104.03 mg/L, 60.67 mg/L, respectively. The validation results showed R2val values of 0.77, 0.8 and 0.77, and RMSEP values of 7.6, 172.37 mg/L and 76.57 mg/L, respectively. A novel approach for the classification of South African red wine cultivars based on unique fluorescent fingerprints was also successful with an overall cross validation score of 0.8. The best classification ability (validation score = 0.93) was shown for the data set containing only fermenting wines for the most widely represented cultivars (>20 samples). This approach may provide a useful tool for authentication and quality control by regulatory bodies.

Untangling the impact of red wine maceration times on wine ageing. A multidisciplinary approach focusing on extended maceration in Shiraz wines.

Phenolic composition of young red wines has been shown to play an important role in their ageing potential. Therefore, the modulation of phenolic extraction during maceration may influence the subsequent phenolic evolution of these wines. The present work aimed to evaluate the impact of three different maceration times on the phenolic levels and evolution observed over time, using spectrophotometric and chromatography methods, and the effect on the aroma, taste, and mouthfeel sensory properties using Projective Mapping. Additionally, grape cell wall deconstruction was monitored during the extended maceration phase by GC-MS and Comprehensive Comprehensive Microarray Polymer Profiling (CoMPP). Our findings demonstrated that longer maceration times did not always correspond to an increase in wine phenolic concentration, although the level of complexity of these molecules seemed to be higher. Additionally, continuous depectination and possible solubilisation of the pectin is observed during the extended maceration which may be influencing the sensory perception of these wines. Maceration time was also shown to influence the evolution of the polymeric fraction and sensory perception of the wines.

The renin-angiotensin-system and left ventricular mass in young adults: the African-PREDICT study.

PURPOSE: Raised blood pressure, with the renin-angiotensin system (RAS) as a central regulatory component, is one of the most important contributors to early development of left ventricular hypertrophy. Factors such as increased age, sex, black ethnicity and a low socio-economic status also contribute to left ventricular remodelling. To better understand early contributors to left ventricular mass, we investigated the relationship between left ventricular mass index (LVMi) and the components of the RAS in young healthy adults while considering ethnicity, sex and socio-economic status. MATERIALS AND METHODS: Black and white women and men (N = 1186) between the ages of 20-30 years were included. By using standard echocardiography, we determined LVMi. Ultra-pressure-liquid chromatography tandem-mass spectrometry (LC-MS/MS) was used to measure the RAS-fingerprint®. RESULTS: Components of the RAS such as plasma renin activity (PRA-S), angiotensin I (Ang I), angiotensin II (Ang II) and aldosterone were suppressed in the black compared to the white group (all p < 0.001). No associations between LVMi and the RAS were evident in the total, black or white groups. With additional grouping according to sex and socio-economic status, inverse associations between LVMi and PRA-S (ß= -0.168;  p = 0.017), Ang I (ß= -0.155; p = 0.028) and Ang II (ß= -0.172; p = 0.015) were found only in low socio-economic black women. CONCLUSION: Despite a suppressed RAS in the black compared to the white group, components of the RAS were not associated with LVMi in this young cohort. The low socio-economic black women of this study population may be vulnerable to future RAS-related increases in left ventricular mass.

The Effect of Different Oak Products Used during Fermentation and Ageing on the Sensory Properties of a White Wine over Time.

The sensorial evaluation of Chenin blanc wine produced with different types of oak wood treatments over time has not been investigated before. The main aim of this study was to assess the sensory profile, using a trained panel, of a South African Chenin blanc wine fermented and matured in old barrels, three types of new oak barrels, two types of oak staves as well as a stainless steel tank over time, which has not been done before. Results indicate mainly separation between the stainless-steel tank/old barrel treatments from the stave and new barrel treatments, with separation between the latter two treatments also being found. More fruity descriptors were used in the stainless-steel tank and old barrel treatments, with oak-related descriptors being used in the stave and new barrel treatments. Separation between among the new barrel and stave treatments were also noted, which was still reflected during bottle maturation. These results indicate that winemakers need to take cognizance of the sensorial differences induced by barrel and stave maturation in white wines and how these evolve over time.

The relationship of blood pressure with uric acid and bilirubin in young lean and overweight/obese men and women: the African-PREDICT study.

Mounting evidence supports the central role of oxidative stress and inflammation in obesity and the development of hypertension. However, most studies focusing on the non-enzymatic antioxidants, such as uric acid and bilirubin, and their relationship with obesity and hypertension were done in older populations with overt cardiovascular disease. The aim of this study was therefore to compare measures of cardiovascular function (blood pressure and arterial stiffness) and non-enzymatic antioxidants (uric acid and bilirubin) between young healthy lean and overweight/obese men and women and to investigate the link between these variables. We grouped 967 men and women (aged 20-30 years) according to body mass index (BMI) categories (lean BMI < 25 kg/m2; overweight/obese BMI ≥ 25 kg/m2). Cardiovascular measurements included 24 h blood pressure and carotid-femoral pulse wave velocity. Serum samples were used to analyse uric acid and bilirubin. Women and men with a BMI ≥ 25 kg/m2 displayed higher 24 h blood pressure (P < 0.001) and uric acid (P ≤ 0.014) than their lean counterparts; lean women showed higher bilirubin (P < 0.001). In multi-variable adjusted regression analyses we found that 24 h systolic blood pressure was independently associated with uric acid (R2 = 0.10; ß = 0.19; P = 0.017) only in overweight/obese women. In lean women a negative association of 24 h systolic blood pressure with bilirubin (R2 = 0.03; ß = -0.14; P = 0.018) was found. No associations were found in men. In conclusion, we found adverse associations between blood pressure and uric acid in young healthy women with increased adiposity, but not in lean women or men.

Matrix effects influencing the perception of 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl acetate (3MHA) in different Chenin Blanc wines by Projective Mapping (PM) with Ultra Flash profiling (UFP) intensity ratings.

Interaction studies are used in sensory evaluation to elucidate the complex influences (additive, masking, synergistic) that various compounds have on the perception of wine aroma. To these, the interactions between the matrix and the compounds of interest add another layer of complexity. Unlike previous interaction studies, the current work used a rapid method, Projective Mapping (PM) coupled with intensity of attributes, to evaluate the interaction effects two thiols (3-mercaptohexan-1-ol, 3MH, and 3-mercaptohexyl acetate, 3MHA) have in various matrices. The matrices used were increasingly complex, from model wine to partially dearomatized neutral Chenin Blanc to commercial wines. The results reflected the effect of the thiols interaction with the matrix more than that between compounds. The methodology proposed for the data handling highlighted possible advantages and shortcomings of the Projective Mapping with intensity approach. The choice of base matrix as well as the sensory method are relevant when studying interaction effects, and are dependent on the desired outcome of the experiment.

Towards on-line monitoring of phenolic content in red wine grapes: A feasibility study.

Spectroscopy techniques to efficiently measure phenolic composition in grape berries may be a suitable analytical practice, provided that robust calibrations are established. A contactless FT-NIR instrument was used for on-line spectral data collection from grapes transported on a conveyor belt system. Spectral data was also collected on static samples using the same NIR instrument. Spectral measurements of crushed berries captured from the conveyor belt system and the use of the homogenate extraction protocol as reference method provided the most accurate prediction models. Values obtained for errors in prediction (RMSEP%) and RPD were 12% and 2.37, 12.3% and 3.37, 7.8% and 3.2, 16.7% and 2.84 for tannins (mg/g) and anthocyanins (mg/g) on a fresh weight basis, total phenols and colour density (AU), respectively. The results observed in this study show the ability of NIR spectroscopy to monitor the phenolic composition of grape berries transported on a conveyor belt system online.

Investigating the relationship between cell wall polysaccharide composition and the extractability of grape phenolic compounds into Shiraz wines. Part II: Extractability during fermentation into wines made from grapes of different ripeness levels.

This study evaluated the relationship between cell wall breakdown, from Shiraz grapes harvested at three different ripeness levels and the colour and phenolics extracted during alcoholic fermentation into wines. Phenolic differences between the ripeness treatments were minimal after » of the fermentation was completed. However, colour and phenolic content were significantly higher in finished wines made from 25°Brix grapes compared to those from 21°Brix and 23°Brix. Levels of grape cell wall polysaccharide deconstruction during fermentation was a determining correlative factor in relation to phenolic extractability. In this context, the de-pectination observed during ripening was found to enhance this deconstruction or "opening-up" of the grape pomace during fermentation, thus increasing the differential extraction of specific polyphenols, especially polymeric polyphenols, into the wines. Additionally, the degree of cell wall deconstruction seemed to play a role in the possible retention and extraction of specific grape proanthocyanidins, depending on their nature and polymer length.

Investigating the relationship between grape cell wall polysaccharide composition and the extractability of phenolic compounds into Shiraz wines. Part I: Vintage and ripeness effects.

Phenolic compounds play an important role in colour stability and sensory properties of red wine. This study evaluated berry skin cell wall composition and how this influences grape and wine phenolics at different ripeness levels (21°Brix, 23°Brix, and 25°Brix) over two consecutive vintages. The vintage effect was highly significant, especially in the pectin fraction of the grape cell walls and affected the concentrations of certain phenolics extracted. The climatic variance between the seasons might have influenced the differences observed in the grape cell wall compositions. Firstly, a higher grape and wine phenolic content, especially in polymeric phenols, was found in 2015 wines. Additionally, grape berry cell walls, especially at the earliest stages of ripening, were found to be more intact in 2015 than in 2016. Thus, a possible relationship was found between the degree of berry intactness, especially for pectin-rich components, and the corresponding phenolic extractability during the winemaking.

Basic In-Mouth Attribute Evaluation: A Comparison of Two Panels.

Astringency is often difficult to evaluate accurately in wine because of its complexity. This accuracy can improve through training sessions, but it can be time-consuming and expensive. A way to reduce these costs can be the use of wine experts, who are known to be reliable evaluators. Therefore, the aim of this work was to compare the sensory results and the panel performance obtained using trained panelists versus wine experts (winemakers). Judges evaluated twelve red wines for in-mouth basic perception (sweet, sour, bitter, astringent, and burning sensation) following the same tasting protocol and with the samples being presented in two different tasting modalities. Panels' performance and relationship between the chemical composition and the sensory perception were investigated. Both panels showed similar consistency and repeatability, and they were able to accurately measure the astringency of the wines. However, the significant correlations between sensory scores and chemical composition varied with the panel and the tasting modality. From our results, we could see that winemakers tended to discriminate better between the samples when the differences were very small.

Chemometric compositional analysis of phenolic compounds in fermenting samples and wines using different infrared spectroscopy techniques.

The wine industry requires reliable methods for the quantification of phenolic compounds during the winemaking process. Infrared spectroscopy appears as a suitable technique for process control and monitoring. The ability of Fourier transform near infrared (FT-NIR), attenuated total reflectance mid infrared (ATR-MIR) and Fourier transform infrared (FT-IR) spectroscopies to predict compositional phenolic levels during red wine fermentation and aging was investigated. Prediction models containing a large number of samples collected over two vintages from several industrial fermenting tanks as well as wine samples covering a varying number of vintages were validated. FT-NIR appeared as the most accurate technique to predict the phenolic content. Although slightly less accurate models were observed, ATR-MIR and FT-IR can also be used for the prediction of the majority of phenolic measurements. Additionally, the slope and intercept test indicated a systematic error for the three spectroscopies which seems to be slightly more pronounced for HPLC generated phenolics data than for the spectrophotometric parameters. However, the results also showed that the predictions made with the three instruments are statistically comparable. The robustness of the prediction models was also investigated and discussed.

Indications of the prominent role of elemental sulfur in the formation of the varietal thiol 3-mercaptohexanol in Sauvignon blanc wine.

Elemental sulfur is a fungicide traditionally used to control Powdery Mildew in the production of grapes. The presence of sulfur residues in grape juice has been associated with increased production of hydrogen sulfide during fermentation, which could take part in the formation of the varietal thiol 3-mercaptohexanol. This work examines whether elemental sulfur additions to Sauvignon blanc juice can increase the levels of sought-after varietal thiols. Initial trials were performed in South Africa and indicated a positive impact of sulfur on the levels of thiols. Further experiments were then carried out with New Zealand Sauvignon blanc and confirmed a positive relationship between elemental sulfur additions and wine varietal thiols. The formation of hydrogen sulfide was observed when the addition of elemental sulfur was made to clarified juice, along with an increase in further reductive sulfur compounds. When the addition of sulfur was made to pressed juice, prior to clarification, the production of reductive sulfur compounds was drastically decreased. Some mechanistic considerations are also presented, involving the reduction of sulfur to hydrogen sulfide prior to fermentation.

Spectrophotometric Analysis of Phenolic Compounds in Grapes and Wines.

Phenolic compounds are of crucial importance for red wine color and mouthfeel attributes. A large number of enzymatic and chemical reactions involving phenolic compounds take place during winemaking and aging. Despite the large number of published analytical methods for phenolic analyses, the values obtained may vary considerably. In addition, the existing scientific knowledge needs to be updated, but also critically evaluated and simplified for newcomers and wine industry partners. The most used and widely cited spectrophotometric methods for grape and wine phenolic analysis were identified through a bibliometric search using the Science Citation Index-Expanded (SCIE) database accessed through the Web of Science (WOS) platform from Thompson Reuters. The selection of spectrophotometry was based on its ease of use as a routine analytical technique. On the basis of the number of citations, as well as the advantages and disadvantages reported, the modified Somers assay appears as a multistep, simple, and robust procedure that provides a good estimation of the state of the anthocyanins equilibria. Precipitation methods for total tannin levels have also been identified as preferred protocols for these types of compounds. Good reported correlations between methods (methylcellulose precipitable vs bovine serum albumin) and between these and perceived red wine astringency, in combination with the adaptation to high-throughput format, make them suitable for routine analysis. The bovine serum albumin tannin assay also allows for the estimation of the anthocyanins content with the measurement of small and large polymeric pigments. Finally, the measurement of wine color using the CIELab space approach is also suggested as the protocol of choice as it provides good insight into the wine's color properties.

Chemical and Sensory Study on the Evolution of Aromatic and Nonaromatic Compounds during the Progressive Oxidative Storage of a Sauvignon blanc Wine.

The effect of repetitive controlled oxidation on the chemical and sensory composition of a fresh and fruity style Sauvignon blanc wine was investigated. Chemical analyses were conducted together with extensive sensory profiling. A decrease in volatile thiols responsible for the fruity nuances and an increase in oxidation-related compounds, such as acetaldehyde, during the course of the oxidation was observed. The wine evolved from a fresh and fruity one to one with slight oxidation and then developed extreme oxidative characteristics. The control samples (no oxygen added) developed a "cooked" character that could indicate the formation of "reductive" compounds in these wines. Conversely, the wines that received a single dose of oxygen did not develop this flavor and were perceived to be fresher and fruitier than the control samples. The color of the wine evolved before the disappearance of the pleasant aroma.