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Med. Sci. Forum, 2021, CAHD 2020

The 1st International e-Conference on Antioxidants in Health and Disease

Online | 1–15 December 2020

Volume Editor:
Mihalis I. Panagiotidis, The Cyprus Institute of Neurology & Genetics (CING), Cyprus

Number of Papers: 25
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Cover Story (view full-size image): This volume presents a collection of contributions that were made to the virtual event: “The 1st International E-Conference on Antioxidants in Health and Disease (CAHD)”. CAHD brought [...] Read more.
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Research

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185 KiB  
Abstract
Evaluation of the Phenolic Profile, Mineral, and Fatty Acid Content and Antioxidant Activity of Black Cumin before and after an In Vitro Simulated Gastrointestinal Digestion
by Johura Ansary
Med. Sci. Forum 2021, 2(1), 9; https://doi.org/10.3390/CAHD2020-08638 - 1 Dec 2020
Viewed by 849
Abstract
Black cumin (N. sativa; Ranunculaceae family) is well known for its numerous beneficial biological effects, while its seed extracts exhibit anti-diabetic, anti-cancer, immunomodulatory, anti-microbial, anti-inflammatory, anti-hypertensive and antioxidant activities [1,2]. This study aimed to evaluate the phenolic and mineral contents, chemical [...] Read more.
Black cumin (N. sativa; Ranunculaceae family) is well known for its numerous beneficial biological effects, while its seed extracts exhibit anti-diabetic, anti-cancer, immunomodulatory, anti-microbial, anti-inflammatory, anti-hypertensive and antioxidant activities [1,2]. This study aimed to evaluate the phenolic and mineral contents, chemical composition and antioxidant activity of methanolic extracts from dried black cumin seed powder. In addition, we evaluated, after gastrointestinal digestion, the effect of phenolic components and their antioxidant activity by utilising an in vitro gastrointestinal digestion process. Black cumin showed high amounts of total phenolic and flavonoid contents (such as dihydroxybenzoic acid and ferulic acid) via HPLC analysis. Six mineral elements (Ca, Cu, Fe, K, Se and Zn) were determined by using coupled plasma mass spectrometry. Twenty-five (25) fatty acids (13 saturated, 7 unsaturated and 5 unsaturated omega fatty acids) were identified by gas chromatography, with linoleic acid being the most abundant. In addition, black cumin methanolic extract presented higher antioxidant capacity measured by DPPH, FRAP and TEAC. Finally, dried black cumin powder was evaluated after gastrointestinal digestion, with results indicating that phenolics, flavonoids and antioxidant capacity increased in the gastric fraction (1.81-, 1.03- and 2.1-fold, respectively) compared with the undigested methanolic extract. Moreover, a higher amount of total phenolic and flavonoid content, as well as a higher total antioxidant capacity were found present in the gastric and elimination fraction than in the bio-accessible fraction (that represent the colon availability). Our results demonstrate a significant reduction in the quantity of phenolic (68%) and flavonoid (95.53%) compounds after gastrointestinal digestion in the bio-accessible fraction, together with a decrease in total antioxidant activity, suggesting that phenolic compounds are responsible for the observed antioxidant activity. Full article
161 KiB  
Abstract
Cruciferous Vegetables-Based Isothiocyanate Compounds as Novel Epigenetic Modulators in Human Malignant Melanoma
by Mihalis I. Panayiotidis
Med. Sci. Forum 2021, 2(1), 10; https://doi.org/10.3390/CAHD2020-08892 - 3 Dec 2020
Viewed by 770
Abstract
Among the various types of dietary agents, isothiocyanates (ITCs) have raised scientific interest with their unique properties against disease development, including modulation of the epigenetic machinery. In the context of malignant melanoma (MM), our research efforts have aimed to understand how ITCs induce [...] Read more.
Among the various types of dietary agents, isothiocyanates (ITCs) have raised scientific interest with their unique properties against disease development, including modulation of the epigenetic machinery. In the context of malignant melanoma (MM), our research efforts have aimed to understand how ITCs induce cell death by interacting with the epigenetic machinery and thus leading to inhibition of tumor growth. For this purpose, we have utilized an in vitro model of human MM consisting of normal keratinocytes, primary and metastatic melanoma as well as non-melanoma epidermoid carcinoma cell lines. In this model, specific ITCs (e.g., sulforaphane (SFN), iberin (IBN) and allyl isothiocyanate (AITC)) were examined for their ability to influence histone acetylation and methylation marks, as a potential epigenetic therapeutic strategy against MM. Overall, we report that all ITCs inhibited MM cell proliferation and influenced acetylation and methylation status of specific lysine residues on H3 and H4 by modulating the expression of various histone acetyl transferases (HATs), histone deacetylases (HDACs) and histone methyl transferases (HMTs) in MM cells. Our data highlight novel insights on SFN, IBN and AITC interaction with components of the histone regulatory machinery, to exert their anticancer action in MM. Full article
148 KiB  
Abstract
Modelling of Protein–Polyphenol Interactions
by Stephen R. Euston
Med. Sci. Forum 2021, 2(1), 22; https://doi.org/10.3390/CAHD2020-08893 - 3 Dec 2020
Viewed by 700
Abstract
The interaction between proteins and polyphenols is known to modify both the bioavailability and bioactivity of dietary polyphenols. Understanding these interactions can facilitate the design of delivery systems for polyphenols in the digestive tract. Molecular modelling of protein–polyphenol and protein–ligand interactions in general [...] Read more.
The interaction between proteins and polyphenols is known to modify both the bioavailability and bioactivity of dietary polyphenols. Understanding these interactions can facilitate the design of delivery systems for polyphenols in the digestive tract. Molecular modelling of protein–polyphenol and protein–ligand interactions in general has long been used as a way to identify small molecule binding sites on proteins. However, these are often used without a careful consideration of the assumptions used and limitations of these methods, and how this affects the accuracy of the predictions. In this paper, two common methods for predicting binding site location and binding energy, molecular dynamics simulation and molecular docking, will be discussed. The simplifications and assumptions implicit in these approaches, as well as ways to improve their predictions will be covered. Full article
175 KiB  
Abstract
Pre-Clinical Investigations of Verbena officinalis L. Tisane Effects against Induced Stress in Wistar Male Rats
by Sonia M. Oliveira, Elsa Dias and Maria de Lourdes Pereira
Med. Sci. Forum 2021, 2(1), 24; https://doi.org/10.3390/CAHD2020-08864 - 30 Nov 2020
Viewed by 879
Abstract
Verbena officinalis or vervain is globally used as an herbal medicine and dietary supplement for anti-depressive and anti-convulsive purposes, as well as to treat inflammatory disorders, skin burns, abrasions and gastric problems. In our exploratory research, we investigated the biochemical, antioxidant and histopathological [...] Read more.
Verbena officinalis or vervain is globally used as an herbal medicine and dietary supplement for anti-depressive and anti-convulsive purposes, as well as to treat inflammatory disorders, skin burns, abrasions and gastric problems. In our exploratory research, we investigated the biochemical, antioxidant and histopathological effects of local V. officinalis infusion in rats previously submitted to chronic physical stress. The animals presented significant alterations in several organs’ ratios; namely the epididymis and brain ratios with p = 0.003 and p = 0.013, respectively. Moreover, tissues such as those of the kidney and liver presented relevant histologic alterations due to experimental conditions. Total protein, creatine kinase (CKI), uric acid (URCA), circulating and hepatic alkaline phosphatase (ALP) and Glutathione S-transferases (GSTs) and glucose levels were statistically different between treated and non-treated animals with p < 0.05. Altogether, biochemical and haematological results indicated significant impacts in antioxidant, lipidic and protein metabolism. Therefore, physical stress and vervain infusion have significant in vivo effects. Chronic stress effects were not counteracted by vervain consumption (e.g., p = 0.5 for hepatic and renal superoxide dismutase (SOD) levels between the different groups). A correlation between histology and the active components in an herbal extract would enable a better evaluation of herbal medicines. Accordingly, further studies of vervain extracts effects are in progress. Full article

Other

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246 KiB  
Proceeding Paper
Comparative Evaluation of the Total Antioxidant Capacities of Plant Polyphenols in Different Natural Sources
by María del Carmen Villegas-Aguilar, María de la Luz Cádiz-Gurrea, David Arráez-Román and Antonio Segura-Carretero
Med. Sci. Forum 2021, 2(1), 1; https://doi.org/10.3390/CAHD2020-08614 - 30 Nov 2020
Viewed by 1236
Abstract
Phenolic compounds during recent decades have been the object of study by the scientific community due to their high and diverse bioactive potential, with their antioxidant capacity being one of their most studied properties since, due to their structure, they have a high [...] Read more.
Phenolic compounds during recent decades have been the object of study by the scientific community due to their high and diverse bioactive potential, with their antioxidant capacity being one of their most studied properties since, due to their structure, they have a high potential to act as effective compounds against oxidative stress. Due to their great structural diversity, these compounds have been classified into at least 10 different classes based on their basic chemical structures. The aim of this work was to carry out the characterization of the main phenolic content and the determination of the antioxidant capacity of four plant matrices. Firstly, Lippia citriodora, Hibiscus sabdariffa, Olea europaea, and Silybum marianum were chosen as natural sources due to their high content of phenolic compounds. After that, a qualitative characterization of the phenolic profiles of the selected plant extracts was performed using high-performance liquid chromatography coupled with mass spectrometry. Finally, the determination of the total content of polyphenols was carried out using the Folin–Ciocalteu method, and the antioxidant capacity using electron transfer methods (FRAP and TEAC) and hydrogen donation (ORAC). After the chromatographic analysis of the phenolic profile of each matrix, it was observed that there are different major phenolic compound families for each plant matrix. Regarding the antioxidant capacity, L. citriodora showed better results for the FRAP assay, while for the TEAC and ORAC assays the highest values were for S. marianum. Full article
809 KiB  
Proceeding Paper
The Nutraceutical Properties of “Pizza Marinara TSG” a Traditional Food Rich in Bioaccessible Antioxidants
by Irene Dini, Luana Izzo, Giulia Graziani and Alberto Ritieni
Med. Sci. Forum 2021, 2(1), 2; https://doi.org/10.3390/CAHD2020-08563 - 30 Nov 2020
Viewed by 1093
Abstract
Italian gastronomy experiences have ever-enhancing fame around the world. This is due to the linkage between taste and salubriousness commonly related to Mediterranean foods. The market proposes many types of pizza to suit all palates. In this work, the antioxidant potential of the [...] Read more.
Italian gastronomy experiences have ever-enhancing fame around the world. This is due to the linkage between taste and salubriousness commonly related to Mediterranean foods. The market proposes many types of pizza to suit all palates. In this work, the antioxidant potential of the “pizza marinara” included in the register of traditional specialties guaranteed (TSG) was determined. An ABTS method evaluated the antioxidant activity of the homogenized pizza. In vitro digestion models estimated the intestinal and gastric bioaccessibility of the main antioxidant compounds (lycopene and phenolics). To our knowledge, this is the first study to provide the content, antioxidant potential, and bioaccessibility of the antioxidants (polyphenols and lycopene) contained in the traditional pizza “marinara TSG”. Our results showed that the “pizza marinara TSG” had a polyphenol concentration, lycopene level, antioxidant activity, and bioaccessibility of phenolic compounds and lycopene which were better than similar pizzas. They confirmed the nutritional importance of traditional preparations and established the functional potential of “pizza marinara TSG” as a food rich in bio-accessible antioxidants. Full article
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207 KiB  
Proceeding Paper
Influence of Organic and Conventional Agricultural Practices on Chemical Profile, In Vitro Antioxidant and Anti-Obesity Properties of Zingiber officinale Roscoe
by Monica Rosa Loizzo, Patrizia Formoso, Mariarosaria Leporini, Vincenzo Sicari, Tiziana Falco and Rosa Tundis
Med. Sci. Forum 2021, 2(1), 3; https://doi.org/10.3390/CAHD2020-08559 - 30 Nov 2020
Cited by 1 | Viewed by 806
Abstract
Zingiber officinale Roscoe (Zingiberaceae), commonly known as ginger. [...] Full article
2218 KiB  
Proceeding Paper
Membrane Proteins of Keratinocyte Protection by Cannabidiol Applied before and after UVB Irradiation
by Sinemyiz Atalay, Agnieszka Gęgotek and Elżbieta Skrzydlewska
Med. Sci. Forum 2021, 2(1), 4; https://doi.org/10.3390/CAHD2020-08603 - 30 Nov 2020
Cited by 1 | Viewed by 1037
Abstract
The continuous increase in daily exposure to ultraviolet radiation, which influences the redox state of skin cells, may contribute to the damage to the structure and function of cellular macromolecules, which favors the search for protective compounds. One promising compound is cannabidiol (CBD), [...] Read more.
The continuous increase in daily exposure to ultraviolet radiation, which influences the redox state of skin cells, may contribute to the damage to the structure and function of cellular macromolecules, which favors the search for protective compounds. One promising compound is cannabidiol (CBD), a phytocannabinoid which has antioxidant and anti-inflammatory properties. Therefore, the aim of this study was to compare the effect of CBD applied after (treatment) as well as before and after (pre-treatment + treatment) keratinocyte irradiation with UVB on the proteomic profile of membrane proteins. The obtained data show that both UVB radiation and CBD treatment significantly modified the proteomic profile of keratinocyte membranes. UVB was shown to dramatically increase the expression of proteins involved in the regulation of cell translation and proliferation (S3a/L13A/L7a ribosomal proteins), calcium ion homeostasis and inflammatory response (S100/S100-A6 proteins) and cellular redox state (peroxireoxin-1). The long action of CBD (pre-treatment + treatment) was more effective in preventing changes caused by UVB, compared to the action of CBD used only after UVB irradiation. The strong activity of CBD applied before and after UVB irradiation suggests that this phytocannabinoid is effective in protecting skin cells against UVB-induced changes, in the keratinocyte proteome. Full article
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480 KiB  
Proceeding Paper
Bioaccessibility of Phenolic Compounds from Cocoa Shell Subjected to In Vitro Digestion and Its Antioxidant Activity in Intestinal and Hepatic Cells
by Silvia Cañas, Miguel Rebollo-Hernanz, Yolanda Aguilera, Vanesa Benítez, Cheyenne Braojos, Silvia M. Arribas and María A. Martín-Cabrejas
Med. Sci. Forum 2021, 2(1), 5; https://doi.org/10.3390/CAHD2020-08612 - 30 Nov 2020
Cited by 2 | Viewed by 1406
Abstract
The cocoa shell is a by-product generated by the cocoa processing industry that could be used as a nutraceutical owing to the significant amounts of bioactive compounds it contains. This work aimed to study the bioaccessibility of phenolic compounds present in the flour [...] Read more.
The cocoa shell is a by-product generated by the cocoa processing industry that could be used as a nutraceutical owing to the significant amounts of bioactive compounds it contains. This work aimed to study the bioaccessibility of phenolic compounds present in the flour (CSF) and an aqueous extract (CSE) from cocoa shells through an in vitro simulated digestion and to assess their antioxidant capacity in vitro by using intestinal and hepatic cell culture models (IEC-6 and HepG2 cells). The bioaccessibility of phenolic compounds was determined using a simulated in vitro digestion model (INFOGEST). Total phenolic compounds (TPC) and antioxidant activity were measured using in vitro techniques. Reactive oxygen species (ROS) were evaluated in IEC-6 and HepG2 cells after t-BOOH stimulation. TPC present in CSE were more bioaccessible than phenolic compounds present in CSF. During digestion, the bioaccessibility of phenolic compounds from CSF fluctuated in the gastric (2.8 mg/g), intestinal (7.6 mg/g), and colonic (5.7 mg/g) phases. Similarly, for the phenolics of CSE, the bioaccessibility increased from 50.6 mg/g in the gastric phase to 53.4 mg/g in the intestinal phase and decreased in the colonic phase to 37.2 mg/g. The in vitro antioxidant capacity followed a similar behavior, increasing throughout the digestion in CSF (8.8- to 10.6-fold) and CSE (6.0- to 7.4-fold). Digested CSF and CSE were not cytotoxic for IEC-6 and HepG2 cells and protected their viability under oxidative stress conditions (93–100%). t-BOOH-induced ROS were prevented by CSF (72–88%) and CSE (81–94%) bioaccessible fractions in both intestinal and hepatic cells. In conclusion, cocoa shells are a source of potentially bioavailable antioxidant phenolic compounds that may protect cells from oxidative stress. Full article
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861 KiB  
Proceeding Paper
Physical-Chemical Properties of the Phenolic Compounds of Humulus lupulus and Aromatic Plant Terpenes: Potential for Use in a Cosmetic Formulation
by Briolanja dos Santos, Olívia R. Pereira, Luís Pedro and Maria João Sousa
Med. Sci. Forum 2021, 2(1), 6; https://doi.org/10.3390/CAHD2020-08558 - 30 Nov 2020
Viewed by 1170
Abstract
Hop (Humulus lupulus L.) are known worldwide as an essential flavor in the beer in-dustry. Their compounds have shown health benefits in terms of phytochemical, pharmacologi-cal and biological profiles, due to their antimicrobial, antioxidant, anti-inflammatory and anti-cancer activities. This study intends to [...] Read more.
Hop (Humulus lupulus L.) are known worldwide as an essential flavor in the beer in-dustry. Their compounds have shown health benefits in terms of phytochemical, pharmacologi-cal and biological profiles, due to their antimicrobial, antioxidant, anti-inflammatory and anti-cancer activities. This study intends to develop a gel formulation incorporating hydroalcoholic extracts of different varieties of Hop such as Cascade, Polaris and spontaneous, from the cones and the vegetative plant parts, in different percentages. The essential oil of Thymus zygis was used as a natural preservative and the analysis of their composition was performed by GC and GC-MS. Additionally, the evaluation of the physical–chemical parameters, stability and capacity to inhibit microbial growth was performed. The pH measurement results of the formulations tend to be adjusted to the natural skin pH. The relative density obtained values of 1 and 0.857. It was found that there were no changes in the phase separation in the centrifugation, vibration, stability and accelerated test. However, changes were observed in the UV-Visible spectra, in texture, consistency and viscosity, and in the color test. In the light test there was phase separa-tion of the samples, which may be related to the manufacturing technique of the formulations. However, anti-aging formulations with phenols from Hop could be developed in order to ob-tain formulations with relevant properties for consumers and the cosmetic industry. Full article
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197 KiB  
Proceeding Paper
Protein-Protein Interaction between Glyoxalase II and Specific Redox Dependent Proteins through S-Glutathionylation Modification
by Laura Cianfruglia, Cristina Minnelli, Andrea Scirè and Tatiana Armeni
Med. Sci. Forum 2021, 2(1), 7; https://doi.org/10.3390/CAHD2020-08615 - 30 Nov 2020
Viewed by 712
Abstract
S-glutathionylation, the reversible formation of a mixed disulfide bridge between a specific cysteine and a glutathione molecule, can occur spontaneously or be catalyzed by enzymes. Glyoxalase II (GloII), using its natural substrate SLG can form specific protein-SSG mixed disulfide leading enzymatic regulation of [...] Read more.
S-glutathionylation, the reversible formation of a mixed disulfide bridge between a specific cysteine and a glutathione molecule, can occur spontaneously or be catalyzed by enzymes. Glyoxalase II (GloII), using its natural substrate SLG can form specific protein-SSG mixed disulfide leading enzymatic regulation of S-glutathionylation. Full article
1466 KiB  
Proceeding Paper
Evaluating the Capacity of Several Antioxidants to Attenuate the Renal Toxicity Induced by Methotrexate Therapy
by Miruna S. Stan and Anca Dinischiotu
Med. Sci. Forum 2021, 2(1), 8; https://doi.org/10.3390/CAHD2020-08597 - 30 Nov 2020
Viewed by 1019
Abstract
Methotrexate (MTX) chemotherapy is often limited by its severe side effects, which include nephrotoxicity. In the continuous search for efficient antioxidants that could ameliorate this toxic condition of MTX, our study aimed to evaluate the efficiency of N-acetyl cysteine (NAC), Trolox methyl ether [...] Read more.
Methotrexate (MTX) chemotherapy is often limited by its severe side effects, which include nephrotoxicity. In the continuous search for efficient antioxidants that could ameliorate this toxic condition of MTX, our study aimed to evaluate the efficiency of N-acetyl cysteine (NAC), Trolox methyl ether (Trolox-Me), and curcumin as potent antioxidants using an in vitro model of MTX-induced toxicity. Human embryonic kidney (HEK293) cells were pre-exposed to different antioxidants for 2 h prior to MTX (5 μM and 50 μM). After 24 h of exposure to MTX, the cell viability and morphology were assessed, activities of antioxidant enzymes and levels of lipid peroxidation were measured by spectrophotometrically methods, and protein expression was determined by Western blotting. Exposure to MTX at concentrations between 1 and 100 μM for 24 h decreased cell viability in a dose-dependent manner and was correlated with the increase in p53 protein expression. All three antioxidants tested have proved that they can inhibit the apoptosis induced by MTX, as revealed by the expression of heat shock proteins (Hsp27, Hsp60, Hsp70 and Hsp90). Pre-treatment of cells with 50 μM of Trolox-Me succeeded to significantly decrease the MTX-induced cell death. The reduction in the activities of glutathione reductase and glutathione S-transferase after MTX incubation was correlated with a low level of GSH, and was attenuated by the pre-incubation with Trolox-Me or curcumin; these antioxidants being able to maintain enough GSH for the reactions of conjugation with MTX metabolites in order to decrease its toxicity. In conclusion, the pre-treatment with curcumin, Trolox-Me or NAC proved extremely effective at blocking MTX toxicity at the concentration investigated in vitro on kidney cells. The results of our study encourage further clinical assessments in order to use these antioxidants in dietary prevention of renal side effects of MTX. Full article
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946 KiB  
Proceeding Paper
Addition of Orange By-Products (Dry Peel) in Orange Jam: Evaluation of Physicochemical Characteristics, Bioactive Compounds and Antioxidant Activity
by Vincenzo Sicari, Monica Rosa Loizzo, Rosa Romeo, Mariarosaria Leporini, Rosa Tundis and Marco Poiana
Med. Sci. Forum 2021, 2(1), 11; https://doi.org/10.3390/CAHD2020-08561 - 30 Nov 2020
Cited by 1 | Viewed by 1451
Abstract
The aim of the present study was to assess the effect of treatment and storage period on the nutritional quality of enriched orange jam during storage for 90 days at 15 and 30 °C. The jam was produced by traditional technique using traditional [...] Read more.
The aim of the present study was to assess the effect of treatment and storage period on the nutritional quality of enriched orange jam during storage for 90 days at 15 and 30 °C. The jam was produced by traditional technique using traditional pectin (TP) and with addition of orange peel (OP). All samples were evaluated for pH, total soluble solids, acidity, total phenolic content, total carotenoid content and antioxidant activity (DPPH and ABTS assay). The mean values at T0 of total soluble solids, titratable acidity and pH of TP jam samples were 63.03 °Brix, 0.60% and 3.51, respectively. While the values for OP jam samples were 65.11 °Brix, 0.61% and 3.68, respectively. The polyphenols ranged from 83.44 to 126.22 mg/100 g for control samples and from 125.12 to 171.02 mg/100 g for enriched jam. As regards temperature storage, the enriched jam stored at 15 °C maintained better analytical parameters than the samples stored at 30 °C. The results obtained from the experimentation show that use of orange peels in orange jam preserved the functional characteristics during storage. Full article
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652 KiB  
Proceeding Paper
Effects of Some New Antioxidants on Apoptosis and ROS Production in AFB1 Treated Chickens
by Consiglia Longobardi, Emanuela Andretta, Vincenzo Romano, Chiara Lauritano, Giuseppina Avantaggiato, Achille Schiavone, Watanya Jarriyawattanachaikul, Salvatore Florio, Roberto Ciarcia and Sara Damiano
Med. Sci. Forum 2021, 2(1), 12; https://doi.org/10.3390/CAHD2020-08640 - 30 Nov 2020
Cited by 4 | Viewed by 1087
Abstract
Aflatoxin B1 (AFB1), the mainly Aspergillus fungi derived mycotoxin, is well known for its carcinogenic effects on liver, and frequently occurs in food supplies, leading to fatal consequences in both farm animals and humans. Poultry, one of the most important segments of agro-industry, [...] Read more.
Aflatoxin B1 (AFB1), the mainly Aspergillus fungi derived mycotoxin, is well known for its carcinogenic effects on liver, and frequently occurs in food supplies, leading to fatal consequences in both farm animals and humans. Poultry, one of the most important segments of agro-industry, has been demonstrated to be extremely sensitive to AFB1 intake, which results in chickens’ low performance, decreased quality of both eggs and meat and a negative economic feedback. Oxidative stress caused by AFB1 plays a crucial role in chickens’ kidney damage by generating lipid peroxidation accompanied by a concomitant increase in the antioxidant enzymes involved in ROS metabolism (NADPH oxidase isoform 4 (NOX4) and its regulatory subunit p47-phox). The aim of the present work was to investigate the benefits of dietary supplementation, in chickens affected by AFB1 mycotoxicosis, using a new Feed additive (FA) containing a mixture of a tri-octahedral Na-smectite with a ligno-cellulose-based material an antioxidant adjuvant. Exposure of AFB1-treated chickens to the feed additive induced a significant down-regulation of both NOX4 and p47-phox genes expression levels. This trend was confirmed by their protein expression, demonstrating the great potential of the FA to counteract oxidative stress. To conclude, these results could open new perspectives in the methods of feeding chickens, using eco-friendly dietary supplements able to reduce AFB1-induced mycotoxicosis and to ameliorate poultry performances. Full article
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1170 KiB  
Proceeding Paper
Chromosome Instability in Asthma an Antioxidant Treatment as a Possible Pharmacological Strategy?
by Cláudia Oliveira, Maria Nogueira, Telma Barbosa, Carmen Cardoso, João Neves, Solange Costa and Beatriz Porto
Med. Sci. Forum 2021, 2(1), 13; https://doi.org/10.3390/CAHD2020-08577 - 30 Nov 2020
Viewed by 1039
Abstract
Asthma is a disease with multiple phenotypes and different degrees of severity. Severe inflammation appears when oxidative stress (OS) overwhelms the antioxidant defense. Knowing the role of OS in the increase of chromosome instability (CI) and, consequently, in cancer predisposition, it is important [...] Read more.
Asthma is a disease with multiple phenotypes and different degrees of severity. Severe inflammation appears when oxidative stress (OS) overwhelms the antioxidant defense. Knowing the role of OS in the increase of chromosome instability (CI) and, consequently, in cancer predisposition, it is important to carry out studies to establish limits for OS harmful effects. The aim of this study was to evaluate OS-related CI in asthma patients and the possible protective effect of antioxidants. For that purpose, spontaneous and OS-induced lymphocyte cultures from patients with mild and severe asthma, and from normal controls, were performed. Antioxidant-enriched cultures from severe patients were posteriorly performed. A hundred metaphases per culture were scored in order to estimate the frequency of CI. Our results showed that lymphocytes from severe patients had increased CI, both in spontaneous and OS-induced cultures. However, in lymphocytes from mild patients there were no differences compared to controls. This suggests that in patients subjected to extreme OS conditions, a genotoxic effect may occur at the cellular level. When lymphocytes from these patients were treated with antioxidants, a decrease in CI was observed. Understanding how CI correlates to asthma patients’ clinical situation may be pivotal to the design of future preventive measures. Full article
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927 KiB  
Proceeding Paper
Cytotoxic and Anti-Proliferative Effects of Fucosterol, Alone and in Combination with Doxorubicin, in 2D and 3D Cultures of Triple-Negative Breast Cancer Cells
by Fernanda Malhão, Alice A. Ramos and Eduardo Rocha
Med. Sci. Forum 2021, 2(1), 14; https://doi.org/10.3390/CAHD2020-08600 - 30 Nov 2020
Cited by 1 | Viewed by 1112
Abstract
Triple-negative breast cancer (TNBC) has the poorest BC prognosis, chemotherapy being the mainstream treatment. Prior studies revealed potential synergistic effects of natural compounds with drugs, such as doxorubicin (Dox), frequently used for TNBC. Fucosterol (Fct), a phytosterol of brown seaweeds, because of its [...] Read more.
Triple-negative breast cancer (TNBC) has the poorest BC prognosis, chemotherapy being the mainstream treatment. Prior studies revealed potential synergistic effects of natural compounds with drugs, such as doxorubicin (Dox), frequently used for TNBC. Fucosterol (Fct), a phytosterol of brown seaweeds, because of its antioxidant and antitumor effects, is one of these promising compounds. Using a TNBC cell line (MDA-MB-231), we aimed to test the effects of Fct alone and in combination with Dox on cell viability and proliferation, in monolayer and three-dimensional (3D) cultures. At this stage of the research, data demonstrated that Fct (5 µM) alone did not affect cell viability and proliferation. In monolayer, Dox (≥1 µM) decreased cell viability and proliferation, while in 3D only cell viability was affected at Dox 5 µM. The combination of Fct/Dox (5/0.1 µM), in monolayer, significantly decreased cell viability and proliferation, differing from the control and both compounds alone; hence, this suggests that Fct enhanced the Dox effect. These promising Fct effects in monolayer were not observed in 3D. We suggest that Fct may increase Dox effects, under certain conditions. Our results corroborate other studies reporting more treatment resistance of cells in 3D culture, reinforcing the need to use more complex models for more realistic drug screening. Full article
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255 KiB  
Proceeding Paper
Total Phenolic and Flavonoid Content and In Vitro Antioxidant Activity of Methanol Extract and Solvent Fractions of Desmodium ramosissimum G. Don
by Uchechukwu S. Ezealigo, Parker E. Joshua, Chidinma P. Ononiwu, Matthias O. Agbo, Rita O. Asomadu and Victor N. Ogugua
Med. Sci. Forum 2021, 2(1), 15; https://doi.org/10.3390/CAHD2020-08594 - 30 Nov 2020
Cited by 4 | Viewed by 1701
Abstract
Oxidative stress has been linked to the pathogenicity of many diseases. This study investigated the total phenolic content (TPC) and total flavonoid content (TFC) of the methanolic extract and solvent fractions (n-hexane, ethyl acetate, n-butanol, and aqueous) of Desmodium ramosissimum [...] Read more.
Oxidative stress has been linked to the pathogenicity of many diseases. This study investigated the total phenolic content (TPC) and total flavonoid content (TFC) of the methanolic extract and solvent fractions (n-hexane, ethyl acetate, n-butanol, and aqueous) of Desmodium ramosissimum using Folin–Ciocalteu and aluminum chloride assays, respectively. The extract and solvent fractions were further appraised for their in vitro antioxidant capacity using total antioxidant capacity (TAC), 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, and ferric reducing antioxidant power (FRAP) methods at varying concentrations of 25–300 µg/mL. Results revealed that ethyl acetate and n-butanol fractions possessed elevated levels of TPC and TFC when compared to other solvent fractions and extracts in a concentration-dependent manner. The ethyl acetate fraction had the highest TPC (532.36 mg GAE/g), TFC (2843.33 mg QE/g), and ferric reducing potential (56.70 mg GAE/g) at 300 µg/mL. In addition, at 300 µg/mL, the TAC (77.33 mg AAE/g) of the n-butanol fraction and its DPPH radical scavenging ability (86.04%) were higher. As shown in this study, organic solvents with different chemical natures are capable of extracting chemical constituents with antioxidant components of different polarities, and D. ramosissimum may also be considered a rich source of natural antioxidants, justifying its pharmacological use in traditional medicine. Full article
1082 KiB  
Proceeding Paper
Redox Behavior of Central-Acting Opioid Tramadol and Its Possible Role in Oxidative Stress
by Uriel Abe Contardi, Mateus Morikawa and Douglas Vieira Thomaz
Med. Sci. Forum 2021, 2(1), 16; https://doi.org/10.3390/CAHD2020-08557 - 30 Nov 2020
Cited by 2 | Viewed by 1429
Abstract
Tramadol (TRA) is a central-acting opioid whose biological activities are achieved by interaction with several bodily receptors such as μ-opioid receptors. Considering that central-acting drugs may promote oxidative stress, which could lead to neurodegeneration, this work reported the investigation of the redox behavior [...] Read more.
Tramadol (TRA) is a central-acting opioid whose biological activities are achieved by interaction with several bodily receptors such as μ-opioid receptors. Considering that central-acting drugs may promote oxidative stress, which could lead to neurodegeneration, this work reported the investigation of the redox behavior of TRA by electrochemical and semi-empirical quantum chemistry approaches (i.e., voltammetry and extended Hückel method—EHM) in order to study TRA pro-oxidant features. Electrochemical results showed that TRA exhibited two anodic peaks, namely: 1a at Ep1a ≈ +0.03 V and 2a at Ep2a ≈ +0.8 V; and a cathodic peak at Ep1c ≈ −0.01 V, whereas the quantum chemistry model suggested that the highest occupied molecular orbital n = 0 (HOMO-0) was associated with the tertiary amine in the TRA molecule, while HOMO-1 and the lowest unoccupied molecular orbital n = 0 (LUMO-0) were associated with the aromatic benzene ring. The findings were then used to propose an electrooxidation pathway according to the observations and compared to the literature, which further offered hints about TRA’s pro-oxidant nature. In conclusion, the work reported herein shows that voltammetric and semi-empirical quantum chemistry approaches can be correlated to investigate the redox behavior of CNS-acting compounds. Full article
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Proceeding Paper
Synthesis and Evaluation of New Nitrone-Based Benzoxazinic Antioxidants
by Stefania Marano, Cristina Minnelli, Giovanna Mobbili, Emiliano Laudadio and Pierluigi Stipa
Med. Sci. Forum 2021, 2(1), 17; https://doi.org/10.3390/CAHD2020-08610 - 30 Nov 2020
Cited by 1 | Viewed by 1074
Abstract
Oxidative stress is often the cause of a wide range of human chronic pathologies including, but not limited to, stroke and cardiovascular and neurodegenerative diseases. In this setting, increasing efforts are currently being devoted to the design and synthesis of new derivatives with [...] Read more.
Oxidative stress is often the cause of a wide range of human chronic pathologies including, but not limited to, stroke and cardiovascular and neurodegenerative diseases. In this setting, increasing efforts are currently being devoted to the design and synthesis of new derivatives with enhanced antioxidant efficacy. Among all the potential molecules of interest, synthetic nitrone spin-traps have attracted a great deal of research attention, particularly due to their dual function as effective inhibitors of oxidative stress and damage and as analytical tools for the detection and characterization of free radicals by means of the electron paramagnetic resonance (EPR) spectroscopy spin trapping technique. In this study, two derivatives of benzoxazinic nitrones (3-aryl-2H-benzo[1,4]oxazin-N-oxides) bearing an electron-withdrawing methyl-acetate group on the benzo moiety (in para and meta positions with respect to the nitronyl function) were synthesized. Their in vitro antioxidant activity was evaluated by means of the α,α-diphenyl-β-picrylhydrazyl radical (DPPH) scavenging assay, and their inhibitory effects on the erythrocyte hemolysis induced by the water-soluble free radical initiator 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH) compared. In addition, EPR was employed to monitor the decay profiles of DPPH to evaluate the kinetic behavior of the different antioxidants tested. Results showed that the presence and the position of the electron-withdrawing methyl-acetate group strongly affects the radical scavenging activity of nitrones. In particular, the newly synthesized para-substituted derivative, when compared to both the meta-substituted isomer and the unsubstituted parent compound, acts as a more effective antioxidant both in cell and cell-free systems. Overall, these results clearly show that the introduction of an electron-withdrawing group on the phenyl moiety significantly increased the antioxidant capacity of benzoxazinic nitrones, thus showing exciting opportunities in the search for new therapeutic agents in the treatment of diseases associated with oxidative stress. Full article
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644 KiB  
Proceeding Paper
Protective Effects of New Antioxidants in OTA-Treated Chicken Kidney
by Emanuela Andretta, Consiglia Longobardi, Martina Laselva, Chiara Lauritano, Giuseppina Avantaggiato, Achille Schiavone, Watanya Jarriyawattanachaikul, Salvatore Florio, Sara Damiano and Roberto Ciarcia
Med. Sci. Forum 2021, 2(1), 18; https://doi.org/10.3390/CAHD2020-08617 - 30 Nov 2020
Cited by 1 | Viewed by 1303
Abstract
Ochratoxin A (OTA) is a mycotoxin which represents an emerging problem for both animal and human health, due to its high presence in feed and foods. Exposure to OTA is associated with oxidative stress-induced nephrotoxicity. Therefore, the identification of new antioxidant or adsorbent [...] Read more.
Ochratoxin A (OTA) is a mycotoxin which represents an emerging problem for both animal and human health, due to its high presence in feed and foods. Exposure to OTA is associated with oxidative stress-induced nephrotoxicity. Therefore, the identification of new antioxidant or adsorbent substances with protective action constitutes one of the main challenges to reduce the negative effects induced by mycotoxins. For this purpose, we investigated the effect of two innovative feed additives, a bio-organoclay (CHS) and a mixture of a tri-octahedral Na-smectite with a ligno-cellulose based material (MIX) alone or in combination with OTA in kidneys of treated chickens. Real-Time PCR analyses for NADPH oxidase 4 (NOX) and p47-phox were performed to evaluate oxidative stress. Our results demonstrated an increase in NOX and p47-phox levels in OTA-treated chickens. Moreover, CHS, more than MIX, was able to reduce OTA-induced toxicity, restoring NOX levels. Taken together, these findings highlight the potential beneficial role of CHS in reverting OTA-induced nephrotoxicity in chickens and could lead to the production of healthier foods with beneficial consequences for human and animal health. Full article
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7 pages, 255 KiB  
Proceeding Paper
Extracts of Different Polarity of Daphne laureola L. as Valuable Source of Antioxidant and Neuroprotective Compounds
by Rosa Tundis, Anna Rita Cappello, Marco Bonesi, Lorenzo Peruzzi and Monica Rosa Loizzo
Med. Sci. Forum 2021, 2(1), 19; https://doi.org/10.3390/CAHD2020-08560 - 30 Nov 2020
Viewed by 1342
Abstract
In this study, Daphne laureola L., a European–Mediterranean species, was investigated for its antioxidant properties using different in vitro bioassays, namely 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), Ferric Reducing Activity Power (FRAP), and β-carotene bleaching assays. The acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activity [...] Read more.
In this study, Daphne laureola L., a European–Mediterranean species, was investigated for its antioxidant properties using different in vitro bioassays, namely 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), Ferric Reducing Activity Power (FRAP), and β-carotene bleaching assays. The acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activity using Ellman’s method was also examined. The aerial parts of D. laureola were subjected to exhaustive and subsequent macerations with solvents at different polarities, such as methanol, dichloromethane, and n-hexane. Dichloromethane extract was the most promising in DPPH and FRAP tests with IC50 values of 32.2 μg/mL and 71.5 μM Fe(II)/g, respectively. Methanol extract showed the most promising inhibition of lipid peroxidation evaluated by β-carotene bleaching test with an IC50 value of 6.6 μg/mL after 30 min of incubation. This extract was the most active against AChE (IC50 value of 56.9 μg/mL). An interesting result was obtained against BChE by the n-hexane extract, with an IC50 value of 49.7 μg/mL. In conclusion, the results suggest that D. laureola may provide a substantial source of phytochemicals, which act as natural antioxidants and acetylcholinesterase inhibitors, and may be beneficial in the treatment of Alzheimer disease. Full article
987 KiB  
Proceeding Paper
High-Fat Diet Promotes a Pro-Inflammatory Environment in Testis and Inhibits Antioxidant Defenses in the Progeny
by Luís Crisóstomo, Romeu A. Videira, Ivana Jarak, Kristina Starčević, Tomislav Mašek, Luís Rato, João F. Raposo, Pedro F. Oliveira and Marco G. Alves
Med. Sci. Forum 2021, 2(1), 20; https://doi.org/10.3390/CAHD2020-08623 - 30 Nov 2020
Viewed by 1075
Abstract
The adoption of high-fat diets (HFDs) is a major contributor to the increasing prevalence of obesity worldwide. Herein, we study the impact of an HFD from early age in testicular physiology and sperm parameters in two generations of mice, with a focus on [...] Read more.
The adoption of high-fat diets (HFDs) is a major contributor to the increasing prevalence of obesity worldwide. Herein, we study the impact of an HFD from early age in testicular physiology and sperm parameters in two generations of mice, with a focus on testicular oxidative status. Mice of the diet-challenged generation (F0; n = 36) were randomly fed after weaning with standard chow (CTRL), a high-fat diet (HFD) for 200 days, or a transient high-fat diet (HFDt) (60 days of HFD + 140 days of standard chow). The offspring generation (F1; n = 36) was obtained by mating normoponderal females with those fed a chow diet 120 days post-weaning. Mice fed an HFD for a lifetime presented impaired insulin tolerance, a trait inherited by their sons. The sons of mice fed HFD inherited decreased catalase activity and displayed lower activities of mitochondrial complexes I and IV. Similar to their progenitors, the sons of HFD mice had a higher prevalence of pinhead and bent neck defects than that of the sons of the CTRL mice. The adoption of HFD impaired testicular antioxidant defenses and mitochondrial function in the progeny, which is detrimental to sperm morphology. Full article
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958 KiB  
Proceeding Paper
Evaluation of Antioxidant Properties of Choloroform Extract of Chasmanthera dependens Roots
by Augustine Madueke, Valentine Nwanelo, Emmanuel Tabansi, Pearl Onoh, Remigus Anichebe, Anayo Okosisi and Assumpta Anosike
Med. Sci. Forum 2021, 2(1), 21; https://doi.org/10.3390/CAHD2020-08606 - 30 Nov 2020
Viewed by 1497
Abstract
Chasmanthera dependens is a medicinal plant with wide application in African traditional medicine for the management of several pathologies. We report the antioxidant properties of the chloroform extract of C. Dependens root (CECDR) as to auspiciously provide scientific information that could explain some [...] Read more.
Chasmanthera dependens is a medicinal plant with wide application in African traditional medicine for the management of several pathologies. We report the antioxidant properties of the chloroform extract of C. Dependens root (CECDR) as to auspiciously provide scientific information that could explain some of the reported roles of the plant in human diseases. In vitro studies assayed for 1,1-diphenyl2-picrylhydrazyl (DPPH) scavenging, hydrogen peroxide scavenging, ferric reducing antioxidant power (FRAP), and total antioxidant capacity (TAC). DPPH radical scavenging activity was concentration dependent with an EC50 of 647.67 μg/mL. CECDR showed a positive hydrogen peroxide scavenging with EC50 of 57.78 ± 2.93 μg/mL relative to ascorbic acid standard (EC50: 90.06 μg/mL). The ferric reducing antioxidant power of CECDR at 15.6 μg/mL was 21.18 ± 0.15 µg gallic acid equivalents (GAE) while that of 1000 μg/mL was 0.03 ± 0.00 µg GAE. CECDR at a concentration 15.6 μg/mL showed a TAC of 15.22 ± 7.81 µg ascorbic acid equivalents (AAE) while that of 1000 μg/mL was 0.03 ± 0.00 µg AAE. In vivo analysis of CCl4-induced Wistar rats showed a significant increase (p < 0.05) in the concentrations of superoxide dismutase, catalase, and glutathione peroxidase in the treated groups relative to the untreated control. In conclusion the observed antioxidant properties of CECDR could be attributed to its rich phytochemical repertoire. Full article
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207 KiB  
Proceeding Paper
Natural Aspirin-Like Compounds from White Willow (Salix alba) Bark Extract Prevent Structural Changes of Human Hemoglobin during In Vitro Non-Enzymatic Glycation and Fructation, Preserving Its Peroxidase and Esterase Activity
by Ionela Cristina Nica, Maria Mernea, Gheorghe Stoian and Anca Dinischiotu
Med. Sci. Forum 2021, 2(1), 23; https://doi.org/10.3390/CAHD2020-08602 - 30 Nov 2020
Viewed by 2263
Abstract
Proteins undergo continuous changes under the action of various intrinsic and extrinsic factors, leading to alteration of several intracellular metabolic pathways and the development of various clinical disorders. Non-enzymatic glycosylation is one of the main factors responsible for the progression of diabetic complications [...] Read more.
Proteins undergo continuous changes under the action of various intrinsic and extrinsic factors, leading to alteration of several intracellular metabolic pathways and the development of various clinical disorders. Non-enzymatic glycosylation is one of the main factors responsible for the progression of diabetic complications and the aging process. Although there are currently many effective therapies in the prevention and treatment of these diseases, in the last decade, there has been an increasing trend of replacing synthetic drugs by natural compounds, in order to reduce the side effects that may occur and the production costs. It is well known that aspirin (acetylsalicylic acid) inhibits the glycation process of serum proteins by acetylating N-terminal amino groups and lysine residues in their structure. Therefore, the main purpose of our research was to analyze the non-enzymatic glycation and fructation process of hemoglobin through spectrometric and electrophoretic techniques, in order to reveal how this process could influence the three-dimensional structure and biological function of the protein, and the effect of some natural aspirin-like compounds on the peroxidase and esterase activity of hemoglobin during fructose and glucose binding. In this way, a preliminary phytochemical characterization of a bark extract of white willow (Salix alba) was performed in order to evaluate the content of total phenolics, flavonoids, and salicylic derivatives, as well as the antioxidant activity. Then, human erythrocytes isolated from whole peripheral blood were incubated with different concentrations of fructose/glucose (10, 50, 100 mM) and S. alba extract for 5, 7, 10 and 14 days. The results obtained from the THz spectra confirmed that fructose was more reactive than glucose, so the glycation process took place more slowly than fructation. Also, the presence of S. alba extract showed an antiglycosylating effect, but not a total inhibition of the glycation process. In addition, enzymatic determinations proved that willow bark extract restored the peroxidase and esterase activities to the control levels. Our data indicated that salicylic compounds can be successfully used as substitutes for aspirin, one of the main synthetic compounds with anti-inflammatory and anti-glycosylating roles. Salicin, salicylic acid and other salicylic compounds possess strong antioxidant properties, which give them the ability to participate in the glycosylation process to block the formation of advanced glycation end products (AGEs). Full article
8 pages, 883 KiB  
Proceeding Paper
Phytochemicals from Cocoa Shell Protect Mitochondrial Function and Alleviate Oxidative Stress in Hepatocytes via Regulation of ERK and PI3K-AKT Pathways
by Miguel Rebollo-Hernanz, Yolanda Aguilera, Maria A. Martin-Cabrejas and Elvira Gonzalez de Mejia
Med. Sci. Forum 2021, 2(1), 25; https://doi.org/10.3390/msf2021002025 - 22 Jul 2021
Viewed by 1885
Abstract
This research aimed to assess the impact of an aqueous extract from the cocoa shell and its major phytochemicals on preventing oxidative stress and mitochondrial dysfunction in hepatocytes using an in vitro model of nonalcoholic fatty liver disease (NAFLD). The phytochemicals from cocoa [...] Read more.
This research aimed to assess the impact of an aqueous extract from the cocoa shell and its major phytochemicals on preventing oxidative stress and mitochondrial dysfunction in hepatocytes using an in vitro model of nonalcoholic fatty liver disease (NAFLD). The phytochemicals from cocoa shell were extracted using water and characterized by UPLC-MS/MS analysis. HepG2 cells were cotreated with either the aqueous extract from cocoa shell (CAE, 20–100 µg mL−1) or 10–50 µmol L−1 of pure theobromine, protocatechuic acid, procyanidin B2, epicatechin, and catechin in the presence or absence of palmitic acid (PA, 500 µmol L−1) to mimic NAFLD conditions in vitro. Biomarkers of mitochondrial function and oxidative stress were evaluated 24 h after the cotreatment in cell supernatants and lysates using chemical, biochemical, and immunochemical techniques. CAE and the phytochemicals therein significantly (p < 0.05) protected mitochondrial content (15–100%) and preserved mitochondrial function, promoting O2 consumption (1.2- to 1.8-fold) and ATP production (1.3- to 2.1-fold). Phytochemicals from cocoa shell significantly (p < 0.05) decreased PA-triggered oxidative stress. The mitochondrial membrane potential was maintained (62–100%), and the production of mitochondrial superoxide (26–100%) and total ROS (17–100%) was abrogated. CAE significantly (p < 0.05) modulated cell signaling pathways associated with ROS production and mitochondrial dysfunction, including an increase in the phosphorylation of ERK1/2 (2.8-fold), protein kinase B (AKT) (2.8-fold), GSK3 (2.3-fold), Raf-1 (1.9-fold), and mTOR (1.7-fold). In conclusion, results suggested that the cocoa shell’s phytochemicals could protect mitochondrial liver function and alleviate oxidative stress by modulating key pathways involved in their regulation. Full article
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