Publications by authors named "Mario Estevez"

109 Publications

Melatonin Modulates the Antioxidant Defenses and the Expression of Proinflammatory Mediators in Pancreatic Stellate Cells Subjected to Hypoxia.

Antioxidants (Basel) 2021 Apr 8;10(4). Epub 2021 Apr 8.

Institute of Molecular Pathology Biomarkers, University of Extremadura, 10003 Caceres, Spain.

Pancreatic stellate cells (PSC) play a major role in the formation of fibrotic tissue in pancreatic tumors. On its side, melatonin is a putative therapeutic agent for pancreatic cancer and inflammation. In this work, the actions of melatonin on PSC subjected to hypoxia were evaluated. Reactive oxygen species (ROS) generation reduced (GSH) and oxidized (GSSG) levels of glutathione, and protein and lipid oxidation were analyzed. The phosphorylation of nuclear factor erythroid 2-related factor (Nrf2), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB), and the regulatory protein nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor-alpha (IκBα) was studied. The expression of Nrf2-regulated antioxidant enzymes, superoxide dismutase (SOD) enzymes, cyclooxygenase 2 (COX-2), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were also studied. Total antioxidant capacity (TAC) was assayed. Finally, cell viability was studied. Under hypoxia and in the presence of melatonin generation of ROS was observed. No increases in the oxidation of proteins or lipids were detected. The phosphorylation of Nrf2 and the expression of the antioxidant enzymes catalytic subunit of glutamate-cysteine ligase, catalase, NAD(P)H-quinone oxidoreductase 1, heme oxygenase-1, SOD1, and of SOD2 were augmented. The TAC was increased. Protein kinase C was involved in the effects of melatonin. Melatonin decreased the GSH/GSSG ratio at the highest concentration tested. Cell viability dropped in the presence of melatonin. Finally, melatonin diminished the phosphorylation of NF-kB and the expression of COX-2, IL-6, and TNF-α. Our results indicate that melatonin, at pharmacological concentrations, modulates the red-ox state, viability, and the expression of proinflammatory mediators in PSC subjected to hypoxia.
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http://dx.doi.org/10.3390/antiox10040577DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8070371PMC
April 2021

Allysine and α-Aminoadipic Acid as Markers of the Glyco-Oxidative Damage to Human Serum Albumin under Pathological Glucose Concentrations.

Antioxidants (Basel) 2021 Mar 17;10(3). Epub 2021 Mar 17.

Meat and Meat Products Research Institute (IPROCAR), Food Technology, University of Extremadura, 10003 Cáceres, Spain.

Understanding the molecular basis of the disease is of the utmost scientific interest as it contributes to the development of targeted strategies of prevention, diagnosis, and therapy. Protein carbonylation is a typical feature of glyco-oxidative stress and takes place in health disorders such as diabetes. Allysine as well as its oxidation product, the α-amino adipic acid (α-AA) have been found to be markers of diabetes risk whereas little is known about the chemistry involved in its formation under hyperglycemic conditions. To provide insight into this issue, human serum albumin was incubated in the presence of FeCl3 (25 μM) and increasing glucose concentrations for 32 h at 37 °C. These concentrations were selected to simulate (i) physiological fasting plasma concentration (4 mM), (ii) pathological pre-diabetes fasting plasma concentration (8 mM), and pathological diabetes fasting plasma concentration (12 mM) of glucose. While both allysine and α-AA were found to increase with increasing glucose concentrations, the carboxylic acid was only detected at pathological glucose concentrations and appeared to be a more reliable indicator of glyco-oxidative stress. The underlying chemical mechanisms of lysine glycation as well as of the depletion of tryptophan and formation of fluorescent and colored advanced glycation products are discussed.
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http://dx.doi.org/10.3390/antiox10030474DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8002732PMC
March 2021

Cross-cultural emotional response to food stimuli: Influence of consumption context.

Food Res Int 2021 04 6;142:110194. Epub 2021 Feb 6.

IPROCAR Research Institute, TECAL Research Group, University of Extremadura, 10003 Cáceres, Spain. Electronic address:

Emotional responses elicited by certain types of food can be influenced by past experiences, frequency of consumption, culture, and other personal preferences. The present research aimed i) to investigate the impact of culture (Brazilian and Spaniard) on consumers' emotional responses and acceptability of different food stimuli, and ii) to explore the influence of evoked contexts. Brazilian (n = 437) and Spanish (n = 397) participants were exposed to three visual food stimuli (image of chocolate, potato chips, and yogurt) in an online survey and reported their emotional responses. Sociodemographic data, liking, and frequency of consumption were also collected. The evoked context in our study were designed and proposed, for each product and culture, based on four dimensions (consumption time, location, social setting, and hungry state). The evoked emotional lexicon was different for each food stimulus and was clearly influenced by the cultural factor. However, there are more similarities between cultures when evaluating the same product category. The evoked contexts were appropriated and influenced the citing frequency of some emotion terms, including positive ones. The most cited emotion terms tended to positively impact product liking ratings, acting as drivers of liking. Consumption level was positively related to liking regardless of cultural interactions for both chocolate and potato chips stimuli. In conclusion, the cultural background demonstrated to be an important impact factor to be considered for understanding the effects of product, consumption occasions, and degree of liking, on emotional responses to foods. These findings offer new possibilities to be explored in marketing messages for interventions or stimuli that guide food choices.
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http://dx.doi.org/10.1016/j.foodres.2021.110194DOI Listing
April 2021

Molecular mechanisms of the disturbance caused by malondialdehyde on probiotic Lactobacillus reuteri PL503.

Microb Biotechnol 2020 Dec 23. Epub 2020 Dec 23.

IPROCAR Research Institute, Food Technology, University of Extremadura, Cáceres, 10003, Spain.

This study aimed to provide insight into the molecular and genetic mechanisms implicated in the responses of Lactobacillus reuteri against the oxidative stress induced by malondialdehyde (MDA) by analysing protein oxidation and assessing the uspA and the dhaT genes. Four experimental groups were evaluated depending on the concentration of MDA added in Man, Rogosa and Sharpe (MRS) broth: Control (L. reuteri), 5 µM (L. reuteri + 5 µM MDA), 25 µM (L. reuteri + 25 µM MDA) and 100 µM (L. reuteri + 100 µM MDA). Three replicates were incubated at 37 °C for 24 h in microaerophilic conditions and sampled at 12, 16, 20 and 24 h. The upregulation of the uspA gene by L. reuteri indicates the recognition of MDA as a potential DNA-damaging agent. The dhaT gene, encoding a NADH-dependent-oxidoreductase, was also upregulated at the highest MDA concentrations. This gene was proposed to play a role in the antioxidant response of L. reuteri. The incubation of L. reuteri with MDA increased the production of ROS and caused thiol depletion and protein carbonylation. L. reuteri is proposed to detoxify pro-oxidative species while the underlying mechanism requires further elucidation.
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http://dx.doi.org/10.1111/1751-7915.13723DOI Listing
December 2020

Impact of 'free-from' and 'healthy choice' labeled versions of chocolate and coffee on temporal profile (multiple-intake TDS) and liking.

Food Res Int 2020 11 19;137:109342. Epub 2020 May 19.

IPROCAR Research Institute, TECAL Research Group, University of Extremadura, 10003 Cáceres, Spain.

Product reformulation for obtaining "free-from" or "healthy choice" versions may confer significant changes in sensory characteristics which could not meet consumer expectations in terms of sensory perception. Therefore, this paper aims to evaluate the influence of formulations of different product categories on their dynamic sensory profile and consumer acceptability. A sensory panel of 23 semi-trained assessors evaluated two product categories, chocolate and milk coffee, using Temporal Dominance of Sensations (TDS) over three consecutive intakes/sips (multiple-intake TDS) on two replications. TDS allowed us to describe the dynamic profile of classic products over the three intakes and to identify differences in the sensory temporal profile compared to their 'healthier choice' reformulated versions. For the chocolate category, increasing the cocoa and the reduction of lactose content mainly influenced the dominance of sweetness, dairy, cocoa, crunchy and bitterness attributes. No sugar addition and absence of caffeine (decaffeinated version) in ready to drink milk coffee beverage modified temporal perception of sweetness, bitterness, coffee flavor and smoothness texture. Higher overall liking averages were associated with the dominant attributes of chocolate with a higher level of cacao and milk coffee decaffeinated. The obtained results of the present work suggest that the multiple-intake TDS technique can be applied for the evaluation of different food categories, helping to develop healthier products and predicting the liking.
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http://dx.doi.org/10.1016/j.foodres.2020.109342DOI Listing
November 2020

Occurrence of wooden breast and white striping in Brazilian slaughtering plants and use of near-infrared spectroscopy and multivariate analysis to identify affected chicken breasts.

J Food Sci 2020 Oct 29;85(10):3102-3112. Epub 2020 Sep 29.

Department of Food Engineering, University of Campinas, Campinas, Sau Paulo, Brazil.

White Striping (WS) and Wooden Breast (WB) are emerging poultry myopathies that occur worldwide, affecting the quality of meat. The aim of this study was to evaluate the occurrence of N, WS, WB, and WS/WB (myopathies combined) in chicken breast from Brazilian commercial plant, comparing (1) inspection based on visual aspect and palpation of Pectoralis major muscle, and (2) identification of these myopathies by near-infrared Spectroscopy (NIRS). Chickens slaughtered at Brazilian commercial plant at four age ranges (4 to 5, 6 to 7, 8 to 9, and 65 weeks) were inspected. Spectral information was acquired using a portable NIR spectrometer, and classification models were performed using and Successive Projection Algorithm-Linear Discriminant Analysis (SPA-LDA) and Soft Independent Modeling of Class Analogy (SIMCA) to distinguish normal and affected muscles. Results showed that occurrence of myopathies was aggravated by age of slaughter, as chicken slaughtered at 4 to 5 and 65 weeks exhibited 13.6 and 95% of myopathies, respectively. Birds slaughtered at 65 weeks showed no occurrence of WB, isolated or combined with WS. It was not possible to differentiate the WB and WS/WB classes; therefore, those samples were grouped (WB+WS/WB). SPA-LDA model showed greater accuracy (92 to 93%) in identifying Normal (N), WS, and WB+WS/WB groups, compared to SIMCA (89 to 91%). It can be concluded that the level of occurrence of myopathies in meat is directly related to the age of slaughter. This study demonstrated that NIRS combined with SPA-LDA model could be used as a tool to detect myopathies in chicken breast. This technique has potential for application in industrial processing lines as an alternative to the traditional methods of identification. PRACTICAL APPLICATION: This study shows that NIRS combined with chemometric techniques can be used to identify chicken breast myopathies in a wide range of ages at slaughter. In addition to being able to discriminate chicken muscles into subclasses, namely, Normal, WS, and WB/WB+WS, this technique has potential for application in industrial processing lines as it is a portable and nondestructive method. This procedure is emphasized as an alternative to the conventional method of identification based on palpation and visual assessment of muscle.
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http://dx.doi.org/10.1111/1750-3841.15465DOI Listing
October 2020

Meat lipids, NaCl and carnitine: Do they unveil the conundrum of the association between red and processed meat intake and cardiovascular diseases?_Invited Review.

Meat Sci 2021 Jan 11;171:108278. Epub 2020 Aug 11.

IPROCAR Research Institute, Food Technology, University of Extremadura, 10003 Cáceres, Spain. Electronic address:

The assessment of the actual contribution of red or processed meat to increasing the risk of suffering cardiovascular diseases (CVD) requires identification of specific harmful components and their underlying pathological mechanisms. In regards to CVD, meat lipids and their oxidation products have been recurrently studied due to their implications on lipid metabolism, hypercholesterolemia, obesity, and risk of suffering vascular events such as stroke. The impact of excess NaCl intake on increasing blood pressure is well-established and processed meat products have been recognized as a major contributor to dietary sodium in developed countries. Recent evidence has also suggested carnitine from red meat, as a precursor for trimethylamine-N-oxide, which has been shown to cause atherosclerosis, may increase the risk of suffering CVD in experimental animals. The present review aims to provide an updated overview, including evidence, controversies and unresolved questions on both the epidemiology and mechanisms relating red and processed meat consumption to CVD.
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http://dx.doi.org/10.1016/j.meatsci.2020.108278DOI Listing
January 2021

Pinpointing oxidative stress behind the white striping myopathy: depletion of antioxidant defenses, accretion of oxidized proteins and impaired proteostasis.

J Sci Food Agric 2021 Mar 12;101(4):1364-1371. Epub 2020 Sep 12.

Institute of Meat and Meat Products (IPROCAR), TECAL Research Group, University of Extremadura, Cáceres, Spain.

Background: This study aimed to investigate the molecular mechanisms involved in the onset of the white striping (WS) myopathy with particular attention to the role of oxidative stress and protein oxidation in the loss of meat quality.

Results: It was found that WS-M (moderate degree; white stripes <1 mm thickness) and WS-S (severe degree; white stripes >1 mm thickness) breast presented higher pH, hardness, redness, lipid, and collagen content, and lower lightness than normal breast. Compared with the latter, WS-S had a more severe loss of protein thiols (70.7% less thiols than in N), reduced activity of antioxidant enzymes such as catalase (23 versus 40 U g ), glutathione peroxidase (0.21 versus 0.54 U g ), and superoxide dismutase (56 versus 73 U g ), and consequently, had greater accretion of thiobarbituric acid reactive substances (0.64 versus 0.22 mg MDAkg muscle), allysine (3.1 versus 1.9 nmol mg protein) and Schiff base structures (645 versus 258 fluorescent units). The analysis of sarcoplasmic proteins revealed that muscles severely affected by the myopathy suffered a chronic impairment of physiological (upregulation of sarcoplasmic reticulum Ca2+ ATPase, sarcalumenin and calsequestrin-2) and metabolic processes (downregulation of pyruvate kinase, creatine kinase, and l-lactate dehydrogenase).

Conclusion: The overexpression of ribonuclease / angiogenin inhibitor 1 and Kelch-like proteins in WS chicken breasts indicates altered protein turnover plausibly mediated by oxidative stress and accumulation of oxidized proteins. © 2020 Society of Chemical Industry.
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http://dx.doi.org/10.1002/jsfa.10747DOI Listing
March 2021

Noxious effects of selected food-occurring oxidized amino acids on differentiated CACO-2 intestinal human cells.

Food Chem Toxicol 2020 Oct 1;144:111650. Epub 2020 Aug 1.

IPROCAR Research Institute, TECAL Research Group, University of Extremadura, 10003, Cáceres, Spain. Electronic address:

The harmful effects of food-occurring oxidized amino acids, namely, aminoadipic acid (AAA), dityrosine (DTYR), L-kynurenine (KN), kynurenic acid (KA) and 3-nitrotyrosine (3NT), were studied on differentiated CACO-2 cells by flow cytometry and quantification of glutathione (GSH), and allysine. Cells were exposed to food-relevant doses (200 μM) of each compound for 4 or 72h and compared to a control (no stimulated cells). All oxidized amino acids induced apoptosis and results indicated that underlying mechanisms depended on the chemical nature of the species. AAA, KN and KA caused ROS generation and severe oxidative stress in 96%, 98% and 89% of exposed cells (77% in control cells), leading to significant GSH depletion and allysine accretion (1.5, 1.5 and 1.6 nmol allysine/mg protein, respectively at 4h; control: 0.22 nmol/mg protein; p < 0.05). DTYR and 3NT induced significant apoptosis to 29% and 25% of cells (control: 16%; p < 0.05) and necrosis to 28% and 26% of cells (control: 23%) at 72h by ROS-independent mechanisms. KN and KA were found to induce a cycle arrest effect on CACO-2 cells. These findings emphasize the potential harmful effects of the intake of oxidized proteins and amino acids and urge the necessity of carrying out further molecular studies.
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http://dx.doi.org/10.1016/j.fct.2020.111650DOI Listing
October 2020

The lysine derivative aminoadipic acid, a biomarker of protein oxidation and diabetes-risk, induces production of reactive oxygen species and impairs trypsin secretion in mouse pancreatic acinar cells.

Food Chem Toxicol 2020 Nov 30;145:111594. Epub 2020 Jul 30.

Institute of Molecular Pathology Biomarkers, University of Extremadura, Cáceres, Spain. Electronic address:

We have examined the effects of α-aminoadipic acid, an oxidized derivative from the amino acid lysine, on the physiology of mouse pancreatic acinar cells. Changes in intracellular free-Ca concentration, the generation of reactive oxygen species, the levels of carbonyls and thiobarbituric-reactive substances, cellular metabolic activity and trypsin secretion were studied. Stimulation of mouse pancreatic cells with cholecystokinin (1 nM) evoked a transient increase in [Ca]. In the presence of α-amoniadipic acid increases in [Ca] were observed. In the presence of the compound, cholecystokinin induced a Ca response that was smaller compared with that observed when cholecystokinin was applied alone. Stimulation of cells with cholecystokinin in the absence of Ca in the extracellular medium abolished further mobilization of Ca by α-aminoadipic acid. In addition, potential pro-oxidant conditions, reflected as increases in ROS generation, oxidation of proteins and lipids, were noted in the presence of α-aminoadipic acid. Finally, the compound impaired trypsin secretion induced by the secretagogue cholecystokinin. We conclude that the oxidized derivative from the amino acid lysine induces pro-oxidative conditions and the impairment of enzyme secretion in pancreatic acinar cells. α-aminoadipic acid thus creates a situation that could potentially lead to disorders in the physiology of the pancreas.
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http://dx.doi.org/10.1016/j.fct.2020.111594DOI Listing
November 2020

Pancreatic stellate cells exhibit adaptation to oxidative stress evoked by hypoxia.

Biol Cell 2020 Oct 12;112(10):280-299. Epub 2020 Jul 12.

Institute of Molecular Pathology Biomarkers, University of Extremadura, Caceres, Spain.

Background Information: Pancreatic stellate cells play a key role in the fibrosis that develops in diseases such as pancreatic cancer. In the growing tumour, a hypoxia condition develops under which cancer cells are able to proliferate. The growth of fibrotic tissue contributes to hypoxia. In this study, the effect of hypoxia (1% O ) on pancreatic stellate cells physiology was investigated. Changes in intracellular free-Ca concentration, mitochondrial free-Ca concentration and mitochondrial membrane potential were studied by fluorescence techniques. The status of enzymes responsible for the cellular oxidative state was analyzed by quantitative reverse transcription-polymerase chain reaction, high-performance liquid chromatography, spectrophotometric and fluorimetric methods and by Western blotting analysis. Cell viability and proliferation were studied by crystal violet test, 5-bromo-2-deoxyuridine cell proliferation test and Western blotting analysis. Finally, cell migration was studied employing the wound healing assay.

Results: Hypoxia induced an increase in intracellular and mitochondrial free-Ca concentration, whereas mitochondrial membrane potential was decreased. An increase in mitochondrial reactive oxygen species production was observed. Additionally, an increase in the oxidation of proteins and lipids was detected. Moreover, cellular total antioxidant capacity was decreased. Increases in the expression of superoxide dismutase 1 and 2 were observed and superoxide dismutase activity was augmented. Hypoxia evoked a decrease in the oxidized/reduced glutathione ratio. An increase in the phosphorylation of nuclear factor erythroid 2-related factor and in expression of the antioxidant enzymes catalytic subunit of glutamate-cysteine ligase, catalase, NAD(P)H-quinone oxidoreductase 1 and heme oxygenase-1 were detected. The expression of cyclin A was decreased, whereas expression of cyclin D and the content of 5-bromo-2-deoxyuridine were increased. This was accompanied by an increase in cell viability. The phosphorylation state of c-Jun NH -terminal kinase was increased, whereas that of p44/42 and p38 was decreased. Finally, cells subjected to hypoxia maintained migration ability.

Conclusions And Significance: Hypoxia creates pro-oxidant conditions in pancreatic stellate cells to which cells adapt and leads to increased viability and proliferation.
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http://dx.doi.org/10.1111/boc.202000020DOI Listing
October 2020

A Chemometric Approach to Establish Underlying Connections between Lipid and Protein Oxidation and Instrumental Color and Texture Characteristics in Brazilian Dry-cured Loin.

Foods 2020 Apr 24;9(4). Epub 2020 Apr 24.

Center for Food Analysis (NAL), Technological Development Support Laboratory (LADETEC), Avenida Horácio Macedo, 1281, Polo de Química, bloco C, Ilha do Fundão, Cidade Universitária, Rio de Janeiro, RJ 21941-598, Brazil.

This study aimed to use chemometrics to evaluate the influence of lipid and protein oxidation on the color and texture characteristics of Brazilian dry-cured loin (Socol, BDL). Upon exploration using hierarchical cluster analysis (HCA), two clusters were formed, indicating that higher water activity (a) was associated with higher lipid and protein oxidation. However, this fact was associated with softening and low color quality (a*, chroma, and cured color). In a more in-depth exploration, using principal component analysis (PCA) for each cluster separately, connections between protein and lipid oxidation were found in high a, as demonstrated by their statistical association. In the same way, relationships between high hardness and carbonyl contents were obtained only in high a. In addition, an overall relationship ( < 0.05) between nondestructive measurements, such as hardness, and destructive methods (malonaldehyde and carbonyl contents) demonstrate that nondestructive techniques can be promising for further studies in the method replacement field. In this study, reasonable explanations of the connections between oxidative damage and quality traits in Socol are provided.
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http://dx.doi.org/10.3390/foods9040536DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7231182PMC
April 2020

Melatonin modulates red-ox state and decreases viability of rat pancreatic stellate cells.

Sci Rep 2020 04 14;10(1):6352. Epub 2020 Apr 14.

Institute of Molecular Pathology Biomarkers, University of Extremadura, Caceres, Spain.

In this work we have studied the effects of pharmacological concentrations of melatonin (1 µM-1 mM) on pancreatic stellate cells (PSC). Cell viability was analyzed by AlamarBlue test. Production of reactive oxygen species (ROS) was monitored following CM-HDCFDA and MitoSOX Red-derived fluorescence. Total protein carbonyls and lipid peroxidation were analyzed by HPLC and spectrophotometric methods respectively. Mitochondrial membrane potential (ψ) was monitored by TMRM-derived fluorescence. Reduced (GSH) and oxidized (GSSG) levels of glutathione were determined by fluorescence techniques. Quantitative reverse transcription-polymerase chain reaction was employed to detect the expression of Nrf2-regulated antioxidant enzymes. Determination of SOD activity and total antioxidant capacity (TAC) were carried out by colorimetric methods, whereas expression of SOD was analyzed by Western blotting and RT-qPCR. The results show that melatonin decreased PSC viability in a concentration-dependent manner. Melatonin evoked a concentration-dependent increase in ROS production in the mitochondria and in the cytosol. Oxidation of proteins was detected in the presence of melatonin, whereas lipids oxidation was not observed. Depolarization of ψ was noted with 1 mM melatonin. A decrease in the GSH/GSSG ratio was observed, that depended on the concentration of melatonin used. A concentration-dependent increase in the expression of the antioxidant enzymes catalytic subunit of glutamate-cysteine ligase, catalase, NAD(P)H-quinone oxidoreductase 1 and heme oxygenase-1 was detected in cells incubated with melatonin. Finally, decreases in the expression and in the activity of superoxide dismutase were observed. We conclude that pharmacological concentrations melatonin modify the redox state of PSC, which might decrease cellular viability.
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http://dx.doi.org/10.1038/s41598-020-63433-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7156707PMC
April 2020

Chicken Combs and Wattles as Sources of Bioactive Peptides: Optimization of Hydrolysis, Identification by LC-ESI-MS and Bioactivity Assessment.

Molecules 2020 Apr 7;25(7). Epub 2020 Apr 7.

Department of Food Engineering, Technology Centre, Federal University of Paraiba, Joao Pessoa 58051-900, Brazil.

The production of bioactive peptides from organic by-waste materials is in line with current trends devoted to guaranteeing environmental protection and a circular economy. The objectives of this study were i) to optimize the conditions for obtaining bioactive hydrolysates from chicken combs and wattles using Alcalase, ii) to identify the resulting peptides using LC-ESI-MS and iii) to evaluate their chelating and antioxidant activities. The hydrolysate obtained using a ratio of enzyme to substrate of 5% (/) and 240 min of hydrolysis showed excellent Fe chelating and antioxidant capacities, reducing Fe and inhibiting 2, 2'-Azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals. The mapping of ion distribution showed that a high degree of hydrolysis led to the production of peptides with / ≤ 400, suggesting low mass peptides or peptides with multiple charge precursor ions. The peptides derived from the proteins of cartilage like Collagen alpha-2(I), Collagen alpha-1(I), Collagen alpha-1(III) and elastin contributed to generation of bioactive compounds. Hydrolysates from chicken waste materials could be regarded as candidates to be used as ingredients to design processed foods with functional properties.
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http://dx.doi.org/10.3390/molecules25071698DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7181024PMC
April 2020

Sulphur amino acids, muscle redox status and meat quality: More than building blocks - Invited review.

Meat Sci 2020 May 12;163:108087. Epub 2020 Feb 12.

College of Food Science and Technology, Nanjing Agricultural University, 210095, PR China.

Advances in food science ineludibly require a profound study of food chemistry fundamentals. Consequently, we need to explore the environment where life and all underlying biological processes occur: the molecular level. The progress made in the last few years in relation to meat protein biochemistry enables revisiting the role that sulphur amino acids (SAA) play as redox-active muscle components. Beyond their participation as building blocks in the de novo synthesis of muscle proteins, protein thiols and other sulphur-containing species are active in controlling metabolic pathways, enzymatic processes, and redox reactions. Supplementation of SAA over nutritional requirements has been found to protect muscle lipids and proteins against oxidation, with subsequent benefits in terms of water-holding capacity, colour stability and sensory properties. As strong nucleophilic compounds, protein thiols establish assorted molecular interactions with other meat and non-meat electrophilic species (i.e. phytochemicals). Those interactions have been found to display remarkable effects on the redox properties, functionality and digestibility of meat proteins. The comprehension of these chemistry fundamentals enables a rationale manipulation of SAA via animal nutrition and/or food formulation to guarantee meat quality and safety and furthermore, develop novel, safer and more nutritious muscle foods, with increased oxidative stability.
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http://dx.doi.org/10.1016/j.meatsci.2020.108087DOI Listing
May 2020

Impact of chicken wooden breast on quality and oxidative stability of raw and cooked sausages subjected to frozen storage.

J Sci Food Agric 2020 Apr 11;100(6):2630-2637. Epub 2020 Feb 11.

Post-Graduate Program in Food Science and Technology, Department of Food Engineering, Technology Center, Federal University of Paraiba, Joao Pessoa, Brazil.

Background: Considering (i) the increased incidence of wooden breast (WB) myopathy in broilers, (ii) the poor meat quality in such breasts and (iii) the lack of studies on the industrial use of WBs; the objective of this study was to investigate the quality of chicken sausages made with WB under frozen conditions. Three formulations of chicken sausage [100% normal (N) breast, 100% WB, and 50% N breast + 50% WB] were considered and raw and cooked (70 °C/10 min) sausages were frozen for 180 days and evaluated for their quality and oxidative stability.

Results: WB formulations exhibited higher pH, moisture (%), and shear force (SF), associated with lower water holding capacity (WHC) and fat (%). During the freezing period, there was a decrease in WHC and redness (a*) and an increase in cooking loss (CL) and SF for all formulations. Additionally, during freezing, thiobarbituric acid-reactive substances values were higher in raw sausages made with N breast than in sausage made with WB probably due to the higher fat content in the former. Results showed that the products did not reach noticeable rancid levels (<2 mg malondialdehyde/kg) and that WB sausage presented great resistance to oxidation, which suggests that WB sausage can be kept frozen at -20 °C for up to 198 days.

Conclusion: Using WBs for the manufacture of chicken sausages subjected to freezing is a viable option given their overall quality and resistance to oxidation. © 2020 Society of Chemical Industry.
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http://dx.doi.org/10.1002/jsfa.10292DOI Listing
April 2020

Benefits of Magnesium Supplementation to Broiler Subjected to Dietary and Heat Stress: Improved Redox Status, Breast Quality and Decreased Myopathy Incidence.

Antioxidants (Basel) 2019 Oct 7;8(10). Epub 2019 Oct 7.

Department of Agricultural and Food Sciences, Alma Mater Studiorum-University of Bologna, 47521 Cesena, Italy.

Poultry is highly sensitive to oxidative reactions. Oxidative reactions have attracted considerable attention from animal and food scientists because of the adverse effects of these reactions on animal welfare, performance and food quality. Despite its implication in multiple biological functions magnesium (Mg) supplementation is typically overlooked in broiler diets. The objective of this study was to evaluate the effect of Mg supplementation (0.3%) using a commercial product (Optibreast) on production parameters, the redox status and meat quality in broilers challenged with dietary (oxidized oil) and heat stress. The incidence of myopathies, namely, wooden breast (WB) and white striping (WS) was also assessed. Mg supplementation had a clear interaction with the absorption/accumulation of Ca in blood and breast muscle but this effect had no negative influence on any of the production parameters under study. Mg supplementation had positive effects on particular meat quality traits such as water holding capacity (WHC) and color. WHC may have other positive effects in turn on relevant sensory traits such as juiciness. Mg supplementation protected against protein oxidation in liver and plasma of broilers. This effect may be related to the increased activity of catalase in such tissues. Mg supplementation reduced the incidence of WS and WB myopathies to almost half the occurrence of such defects in animals fed a control diet. Further studies with a larger number of animals and the application of advanced proteomic/metabolomic tools are required to (1) corroborate the positive influence of Mg on myopathy incidence and (2) identify the underlying molecular basis of the proposed mechanisms.
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http://dx.doi.org/10.3390/antiox8100456DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6826508PMC
October 2019

Increased markers of cardiac vagal activity in leucine-rich repeat kinase 2-associated Parkinson's disease.

Clin Auton Res 2019 12 23;29(6):603-614. Epub 2019 Aug 23.

Facultad de Biología, Universidad de La Habana, Calle 25 No. 455, Vedado, Plaza de la Revolución, 10400, La Habana, Cuba.

Purpose: Cardiac autonomic dysfunction manifests as reduced heart rate variability (HRV) in idiopathic Parkinson's disease (PD), but no significant reduction has been found in PD patients who carry the LRRK2 mutation. Novel HRV features have not been investigated in these individuals. We aimed to assess cardiac autonomic modulation through standard and novel approaches to HRV analysis in individuals who carry the LRRK2 G2019S mutation.

Methods: Short-term electrocardiograms were recorded in 14 LRRK2-associated PD patients, 25 LRRK2-non-manifesting carriers, 32 related non-carriers, 20 idiopathic PD patients, and 27 healthy controls. HRV measures were compared using regression modeling, controlling for age, sex, mean heart rate, and disease duration. Discriminant analysis highlighted the feature combination that best distinguished LRRK2-associated PD from controls.

Results: Beat-to-beat and global HRV measures were significantly increased in LRRK2-associated PD patients compared with controls (e.g., deceleration capacity of heart rate: p = 0.006) and idiopathic PD patients (e.g., 8th standardized moment of the interbeat interval distribution: p = 0.0003), respectively. LRRK2-associated PD patients also showed significantly increased irregularity of heart rate dynamics, as quantified by Rényi entropy, when compared with controls (p = 0.002) and idiopathic PD patients (p = 0.0004). Ordinal pattern statistics permitted the identification of LRRK2-associated PD individuals with 93% sensitivity and 93% specificity. Consistent results were found in a subgroup of LRRK2-non-manifesting carriers when compared with controls.

Conclusions: Increased beat-to-beat HRV in LRRK2 G2019S mutation carriers compared with controls and idiopathic PD patients may indicate augmented cardiac autonomic cholinergic activity, suggesting early impairment of central vagal feedback loops in LRRK2-associated PD.
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http://dx.doi.org/10.1007/s10286-019-00632-wDOI Listing
December 2019

Malondialdehyde interferes with the formation and detection of primary carbonyls in oxidized proteins.

Redox Biol 2019 09 20;26:101277. Epub 2019 Jul 20.

IPROCAR Research Institute, Faculty of Veterinary, University of Extremadura, 10003, Cáceres, Spain; Heart Clinical Unit, Virgen de la Victoria University Clinic Hospital. Institute of Biomedical Research in Malaga. IBIMA. CIBERCV. University of Málaga, Málaga, Spain.

Carbonylation is one of the most remarkable expressions of the oxidative damage to proteins and the DNPH method the most common procedure to assess protein oxidation in biological samples. The present study was elicited by two hypotheses: i) is malondialdehyde, as a reactive dicarbonyl, able to induce the formation of allysine through a Maillard-type reaction? and ii) to which extent does the attachment of MDA to proteins interfere in the assessment of protein carbonyls using the DNPH method? Human serum albumin (HSA), human hemoglobin (HEM) and β-lactoglobulin (LAC) (5 mg/mL) were incubated with MDA (0.25 mM) for 24 h at 37 °C (HSA and HEM) or 80 °C (LAC). Results showed that MDA was unable to induce oxidative deamination of lysine residues and instead, formed stable and fluorescent adducts with proteins. Such adducts were tagged by the DNPH method, accounting for most of the protein hydrazones quantified. This interfering effect was observed in a wide range of MDA concentrations (0.05-1 mM). Being aware of its limitations, protein scientists should accurately interpret results from the DNPH method, and apply, when required, other methodologies such as chromatographic methods to detect specific primary oxidation products such as allysine.
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http://dx.doi.org/10.1016/j.redox.2019.101277DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6669345PMC
September 2019

The impaired quality of chicken affected by the wooden breast myopathy is counteracted in emulsion-type sausages.

J Food Sci Technol 2019 Mar 13;56(3):1380-1388. Epub 2019 Feb 13.

IPROCAR Research Institute, Tecal Research Group, University of Exremadura, 10003 Caceres, Spain.

The aim of this study was to evaluate the suitability of using chicken meat affected by wooden breast (WB) myopathy in the production of chicken sausages. Compare the technological and sensory properties of such sausages were compared with those produced from normal (N) breast meat. Three types of chicken sausages were elaborated: 100% containing N chicken meat, 100% of WB chicken meat and 50% N/50% of WB meat. The WB chicken meat presented higher values for pH, L*, moisture, cooking loss, shear force, hardness, chewiness, adhesiveness and gumminess; while WHC and protein content were higher for N chicken meat. N and WB chicken sausages presented similar values of WHC, a*, b* color values, protein content and TBARS. QDA indicated no sensory differences between the three sausage formulations, so did the acceptability and purchase intention. Therefore, WB chicken meat may be used to produce chicken sausages combined or not with N chicken meat. Further studies, however, may be required to investigate the nutritional value and digestibility of WB meat and derived products.
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http://dx.doi.org/10.1007/s13197-019-03612-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6423235PMC
March 2019

Collapse of the endogenous antioxidant enzymes in - broiler thigh muscles triggers oxidative stress and impairs water-holding capacity.

J Food Sci Technol 2019 Mar 13;56(3):1371-1379. Epub 2019 Feb 13.

4TECAL Research Group, IPROCAR Research Institute, University of Extremadura, Cáceres, Spain.

This study was conducted to investigate the effect of the collapse of the endogenous antioxidant enzymes, namely, catalase (CAT), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) in - (PM) chicken thigh muscles on the extent of lipid and protein oxidation and the functionality of the muscle in terms of water-holding. To fulfil this objective, the samples were divided into two treatments: one group of muscles (n = 8) was subjected to delay cooling (DC) (at ~ 37 °C for 200 min PM) and then stored at 4 °C for 24 h. The second group (n = 8) was subjected to a normal cooling (NC): samples were immediately chilled at 4 °C for 24 h. DC samples presented a decrease in 16% of CAT, 25% GSH-Px and 20% SOD activity in relation to NC. Consistently, an increase of 36% of total carbonyl, 15% of Schiff bases and 27% of TBA-RS and 14% of tryptophan depletion was observed in DC samples, as compared to NC. The results suggested that DC challenged muscles to struggle against oxidative reactions, consuming endogenous antioxidant defenses and causing protein and lipid oxidation which in turn affect the quality and safety of chicken meat. These results emphasize the role of PM oxidative stress on chicken quality and safety. Antioxidant strategies like fast cooling may be combined with others (dietary antioxidants) to preserve chicken quality against oxidative stress.
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http://dx.doi.org/10.1007/s13197-019-03611-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6423192PMC
March 2019

Resveratrol protects Lactobacillus reuteri against HO- induced oxidative stress and stimulates antioxidant defenses through upregulation of the dhaT gene.

Free Radic Biol Med 2019 05 23;135:38-45. Epub 2019 Feb 23.

Meat and Meat Products Research Institute, Food Technology, University of Extremadura, 10003, Cáceres, Spain. Electronic address:

Understanding of the mechanisms implicated in the protective role of probiotic bacteria is of the utmost scientific interest. This study provides original insight into the genetic and molecular basis of the responses of Lactobacillus reuteri PL503 against hydrogen peroxide (HO)-induced oxidative stress. Six experimental groups were considered depending on the addition and concentration of HO and resveratrol: 1. CONTROL (L. reuteri in MRS broth); 2. HO (L. reuteri in MRS broth + 0.5 mM HO); 3. LRES (L. reuteri in MRS broth + 20 μM resveratrol); 4. HRES (L. reuteri in MRS broth + 100 μM resveratrol); 5. HO-LRES (L. reuteri in MRS broth + 0.5 mM HO + 20 μM resveratrol); 6. HO-HRES (L. reuteri in MRS broth + 0.5 mM HO + 100 μM resveratrol). Three replicates were incubated at 37 °C for 24 h in microaerophilic conditions sampled at 12, 16, 20 and 24 h. The NADH-dependent-oxidoreductase encoded by the dhaT gene is a plausible candidate to be strongly implicated in the antioxidant response of L. reuteri. Resveratrol (100 μM) is found to protect L. reuteri against protein carbonylation plausibly through various mechanisms including direct scavenging of reactive oxygen species (ROS), upregulation of the dhaT gene and promoting the synthesis of sulfur containing compounds. The hypothesis formulated on the ability of L. reuteri to detoxify HO and its underlying mechanism needs to be clarified. Furthermore, the consequences of protein carbonylation as a reflection of oxidative damage to bacteria and its role in the responses of bacteria to oxidative stress need to be further investigated.
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http://dx.doi.org/10.1016/j.freeradbiomed.2019.02.023DOI Listing
May 2019

Intake of Oxidized Proteins and Amino Acids and Causative Oxidative Stress and Disease: Recent Scientific Evidences and Hypotheses.

J Food Sci 2019 Mar 4;84(3):387-396. Epub 2019 Feb 4.

Depart. of Animal and Food Sciences, Univ. of Kentucky, Lexington, KY 40546-0215, U.S.A.

The exposure to reactive oxygen species (ROS) is an inevitable consequence of living in an aerobic world. The species contribute to the occurrence of oxidative stress in humans in which an uncontrolled production of ROS exceeds the endogenous antioxidant defences leading to the oxidative damage to essential cellular components, such as lipids, proteins, and DNA. The influence of diet on the modulation of the systemic redox status is recognized and, while some dietary components are found to be protective (that is, fruits and vegetables), others are recognized as pro-oxidants (that is, processed meat and other animal-source protein foods). Oxidized proteins and amino acids are potential promoters of luminal and postprandial oxidative stress; preliminary studies have actually reported noxious effects of these species in cultured cells and in experimental animals. However, the underlying pathological mechanisms remain poorly understood. The application of advanced methodological approaches based on mass spectrometric technologies and OMICS disciplines has enabled the elucidation of the molecular basis of the pathological effects of dietary oxidized proteins and amino acids. The present review collects the most recent evidences of the health risks of dietary protein oxidation and proposes reasonable hypotheses and future perspectives on the field.
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http://dx.doi.org/10.1111/1750-3841.14460DOI Listing
March 2019

Emotional responses to the consumption of dry-cured hams by Spanish consumers: A temporal approach.

Meat Sci 2019 Mar 22;149:126-133. Epub 2018 Nov 22.

Institute of Meat and Meat Products (IPROCAR), University of Extremadura, Avd. Universidad s.n., Cáceres, Spain.

The objective of this study was to describe for the first time the emotional responses during the consumption of three types of Spanish dry-cured hams, namely, 'Iberian', 'Serrano' and 'Curado'. Moreover, dynamic sensory techniques such as Temporal Dominance of Emotions and Temporal Dominance of Sensations were applied to identify the dominant emotions and sensory attributes perceived during ham consumption. >50 emotional terms were generated during sample consumption including positive, negative and neutral terms. Noticeable differences were detected among the three types of dry-cured ham for their emotional profiles. The most dominant emotions evoked by Iberian dry-cured hams were "intense", "authentic" and "pleasant" and could be associated with the domination of cured flavour and juiciness, with both attributes being closely related to their high intramuscular fat content. However, in Serrano and Curado dry-cured hams, the domination of saltiness attribute seemed to contribute to evoking negative emotions as "ordinary", "indifferent" and "dissatisfied". Finally, high duration of the domination of positive emotions as "desirable", "intense", "pleasant" and "authentic" contributed to high overall liking scores.
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http://dx.doi.org/10.1016/j.meatsci.2018.11.015DOI Listing
March 2019

Formation of allysine in β-lactoglobulin and myofibrillar proteins by glyoxal and methylglyoxal: Impact on water-holding capacity and in vitro digestibility.

Food Chem 2019 Jan 25;271:87-93. Epub 2018 Jul 25.

IPROCAR Research Institute, TECAL Research Group, University of Extremadura, 10003 Cáceres, Spain. Electronic address:

The ability of α-dicarbonyls, glyoxal (GO) and methyl-glyoxal (MGO) (2 M), to induce the formation of allysine in β-lactoglubulin (LAC), and myofibrillar proteins (MP) (2 mg/mL) during incubation at 80 °C for 48 h, was studied. Both GO and MGO induced the formation of allysine in all tested proteins with GO being more reactive (23.8 and 8.6 nmoles/mg protein in LAC and MP respectively after 6 h) than MGO (2.6 and 3.1 nmoles/mg protein at the same sampling point). LAC seemed to be more susceptible to the glycation reactions than MP. The concentration of allysine decreased at 24 h along with a concomitant increase of advanced-glycation end-products suggesting that allysine may be involved in the formation of fluorescent adducts. The water-holding capacity and trypsin-chymotrypsin digestibility of the proteins decreased during the incubation assay. The mechanisms by which α-dicarbonyls-mediated carbonylation likely influenced the impairment of such protein properties are thoroughly discussed.
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http://dx.doi.org/10.1016/j.foodchem.2018.07.167DOI Listing
January 2019

Response to Lewis A: Reconciling the Case of Jahi Mcmath.

Neurocrit Care 2018 12;29(3):521-522

The National Institute for Brain and Rehabilitation Sciences, Nazareth, Israel.

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http://dx.doi.org/10.1007/s12028-018-0602-0DOI Listing
December 2018

Further insight into the role of Ca in broiler pale, soft and exudative-like (PSE) meat through the analysis of moisture by TGA and strong cation elements by ICP-OES.

J Food Sci Technol 2018 Aug 19;55(8):3181-3187. Epub 2018 Jun 19.

1Graduate Program in Animal Science, Department of Animal Science, Agricultural Sciences Center, Londrina State University, Londrina, Paraná Brazil.

This study evaluated moisture content of broiler breast PSE meat by thermal gravimetric analysis and strong cation elements by inductively coupled plasma optical emission spectrometry. The weight changes were monitored within the temperature interval 9-750 °C and the most severe changes were observed at 9-160 °C (1A). This remarkable transition was attributed to water loss of about 64.74% in normal samples while PSE samples lost about 61.16%. There was, therefore, approximately 4.0% lower moisture in PSE meat samples in relation to normal meat. The analysis of cation elements showed significant differences (< 0.05), with normal samples having 13.0% higher ∑[Ca] + [Mg] + [Na]+[K] than PSE meat samples while [Ca] in PSE meat was 10.81% higher than in the normal counterparts. We provided further evidence of the role of Ca excess within the muscle sarcomere during the onset of PSE meat.
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http://dx.doi.org/10.1007/s13197-018-3246-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6046006PMC
August 2018

Benefits of wine-based marination of strip steaks prior to roasting: inhibition of protein oxidation and impact on sensory properties.

J Sci Food Agric 2019 Feb 27;99(3):1108-1116. Epub 2018 Sep 27.

Department of Animal Production and Food Science, IPROCAR Research Institute, TECAL Research Group, University of Extremadura, Cáceres, Spain.

Background: The objective of this study was to evaluate the impact of red wine-based marination on the oxidative stability and overall quality of roasted beef strip steaks. Four treatments were considered, according to the type of wine (300 mL dealcoholized wine/kilogram meat): 'Cabernet Sauvignon', 'Tempranillo', 'Isabel' (ISA), and a control. The formation of potentially harmful protein oxidation products during roasting, including protein carbonyls and dityrosines, was inhibited by bioactive components of the wine.

Results: ISA marinades were particularly resistant to protein oxidation, which could be due the particular composition of this wine in phenolic compounds. Wine-based marination was also effective in controlling the formation of lipid-derived volatile compounds, such as hexanal, octane-2,5-dione, and heptan-2-one, which led to a reduced perception of rancidity by panelists. Additionally, wines contributed to spicing roasted beef with wine-derived flavors from esters, alcohols, and lactones.

Conclusions: Hence, marination may be a feasible means to alleviate the potential negative effects that oxidative reactions cause to meat proteins, improve beef quality, and diversify beef cuts into a variety of safer and more flavored meat products. Among wines, ISA appeared to be most promising in terms of antioxidant protection; however, the limited consumer acceptance of steaks treated with this wine may be regarded as a drawback to be sorted out in future studies. © 2018 Society of Chemical Industry.
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http://dx.doi.org/10.1002/jsfa.9278DOI Listing
February 2019