This study investigates the curative response of ethanol extract of Cassia sieberiana leaves (EECSL) on testosterone-induced benign prostatic hyperplasia (BPH). Thirty rats (120-160 g) were grouped into six of five rats each. Group 1 was the normal control; groups 2-6 were induced with testosterone (5 mg/kg body weight) for twenty-one days following standard procedures. Group 2 serves as BPH-control, groups 3 received finasteride while 4-6 received orally 100, 300, and 500 mg/kg of EECSL respectively for 14 days. Biochemical and BPH indices were analyzed. Oral administration of EECSL showed a decrease in prostate weight, Prostate specific antigen (PSA), testosterone and dihydrotestosterone (DHT), triacylglycerol (TAG), total cholesterol (TC), low density lipoprotein (LDL), malondialdehyde (MDA), aspartate aminotransferase (AST) alanine aminotransferase (ALT) whereas an elevation in High density lipoprotein (HDL), GSH and likewise antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) activities were recorded. The histology photomicrograph showed that EECSL restored lesions of testosterone induction in the treated groups. Our study reveals that EECSL exerts potent on BPH, hypolipidaemic, and antioxidant potentials and justifies the claims by the traditional doctors in BPH management.

Metabolic syndrome (MS) is a major global public health concern with its prevalence rising steadily. Plants contain variety of bioactive compounds (phytochemicals) of therapeutic benefits which have proven effective in management and treatment of several human and animal diseases. This study is therefore aimed at evaluating the effect of diet formulations of Gongronema latifolium leaf (GLL), Cnidoscolus aconitifolius leaf (CAL) and Moringa oleifera leaf (MOL) on some biochemical parameters of experimentally-induced MS in male albino rats. Adult Wistar male rats (forty-eight) weighing between 180-210 g were randomly grouped into eight groups of six rats each. Group 1 received normal diet. MS was induced in experimental rats (Groups 2 – 8) with high fat high carbohydrate (HFHC) diet for eight weeks, after which group 2 was fed normal rat diet (untreated), while groups 3 to 8 was treated with diets formulated with GLL, CAL, MOL for another eight weeks. The dose of the plants used for feed formulation was 10%, which is 100 g of plant in 1000 g of the total formulated diet for each treatment. Antioxidant status, liver marker enzymes, serum lipid profile and obesity indices were evaluated using standard methods. Superoxide dismutase and catalase activities significantly (p < 0.05) increased in the treatment groups. Treatment with the herbs showed mild Kupffer cell activation reversing periportal hepatitis induced after the consumption of HFHC diet. Significant (p < 0.05) decrease in total cholesterol, triacylglycerols and body weight gain of the treated groups were observed, while high density lipoprotein significantly (p < 0.05) increased compared to the untreated group. Results from the study indicate that GLL, CAL, and MOL have therapeutic potentials that could be useful in the management of metabolic syndrome components.

Oxidative stress poses a risk to human health and has been linked to the pathogenicity of a myriad of diseases. This study investigated the total phenolics content (TPC) and total flavonoids content (TFC) of Desmodium ramosissimum methanol extract and its solvent fractions (n-hexane, ethyl acetate, n-butanol, and aqueous) using Folin-Ciocalteu and aluminum chloride assays respectively. Furthermore, the extract and solvent fractions were appraised for their in vitro antioxidant capacity using relevant assays: total antioxidant capacity (TAC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and ferric reducing antioxidant power (FRAP) methods all at varying concentrations of 25, 50, 100, 200, 250 and 300 µg/ml. Results revealed that ethyl acetate and n-butanol fractions possess higher levels of TPC and TFC when compared to other solvent fractions and extract in a concentration-dependent manner. The ethyl acetate fraction had the highest TPC (532.36 mgGAE/g of plant extract), TFC (2843.33 mgQE/g of plant extract) and exhibited the highest ferric reducing potential (56.70 mgGAE/g of plant extract) at 300 µg/ml. Also, at 300 µg/ml, the TAC (77.33 mgAAE/g of plant extract) 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. The study also demonstrates that D. ramosissimum may be considered a rich source of natural antioxidants justifying its pharmacological use in traditional medicine for ameliorating diseases.

Oxidative stress has been implicated in many diseases and aging. Electrolysis of water produces electrolyzed-reduced water (ERW) rich in hydrogen molecules and hydrogen atoms (active hydrogen) near the cathode, both of which have been shown to contribute to reduced oxidative stress and improve antioxidant potential by scavenging reactive oxygen species (ROS). We investigated the effects of drinking ERW on biomarkers of oxidative stress and health-related indices in healthy adults at the workplace.

This study was a randomized, double-blind, placebo-controlled clinical trial. Sixty-five participants were allocated into two groups. Of these, 61 received intervention (32 ERW and 29 MW [mineral water]), and were instructed to drink 1.5 L/day of ERW or MW for eight weeks. Biomarkers of oxidative stress and health-related indices were assessed at baseline, four, and eight weeks.

Fifty-three subjects completed the study. Of the primary outcome variables assessed, a significant interaction between the group and time was shown in the diacron-reactive oxygen metabolites (d-ROMs) and biological antioxidant potential, with d-ROMs levels in the ERW group significantly decreased at eight weeks compared with those in the MW group. Among the secondary outcome variables, total, visceral, and subcutaneous fat mass showed a significant change at different time points, with a significant interaction observed between the group and time.

Drinking ERW daily could be suggested as effective, sustainable, and innovatively simple lifestyle modification in healthy adults to reduce oxidative stress, increase antioxidant potential, and decrease fat mass.

Methotrexate (MTX) chemotherapy is often limited by its severe side effects which include nephrotoxicity. In the continuous search of efficient antioxidants that could ameliorate MTX toxicity, 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 before MTX treatment. After 24 hours of exposure to MTX, cell viability and morphology were assessed. Activities of antioxidant enzymes and levels of lipid peroxidation were measured by spectrophotometric methods, and protein expression was determined by Western blotting.

Exposure to MTX at concentrations between 1 and 100 μM for 6 and 24 h decreased cell viability in a dose-dependent manner and was correlated with the increase in p53 protein expression. All three antioxidants tested were able to inhibit MTX-induced apoptosis, 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 treatment was correlated with a reduced level of GSH, and was attenuated by the pre-incubation with Trolox-Me or curcumin. These antioxidants were 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 was significantly effective in 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.

The development of innovative dip-coating technologies for surface functionalization has been a very active issue over the past decade following the discovery of the adhesion properties of polydopamine, a eumelanin-like material. New opportunities have derived from the discovery that hexamethylenediamine (HMDA) markedly enhances film deposition from a variety of catechol, including the key eumelanin precursor 5,6-dihydroxyindole (DHI). The remarkable antioxidant properties of synthetic eumelanins from the other main melanogenic precursor 5,6-dihydroxyindole-2-carboxylic acid (DHICA) and its methyl ester (MeDHICA) have recently been described. In this work the film forming properties of MeDHICA melanin generated in the presence of HMDA or other diamines/monoamines and the antioxidant activity of the resulting films were investigated. Further to a systematic investigation, the most promising results were obtained running the aerobic oxidative polymerization of MeDHICA in aqueous buffers at pH 9.0 at 1 mM in the presence HMDA at 1:1.5 molar ratio. Under these conditions a dark yellow pigment is formed over 24 h exhibiting good film forming properties on different supports. HPLC analysis of the film solubilized in organic solvents indicated a mixture of oligomers of MeDHICA up to hexamers. Further polymerization of the film was obtained by exposure to ammonia vapors. The films showed high antioxidant activities in the 2,2-diphenyl-1-picrylhydrazyl and Ferric Reducing Antioxidant Power assays. Biocompatibility of MeDHICA/HMDA films was assessed on HaCat cells.

The triple-negative breast cancer (TNBC) has the poorest BC prognosis, as patients cannot benefit from target therapies, being chemotherapy the mainstream treatment. Therefore, the search for new drugs/drug adjuvants to overcome drug resistance or reduce their toxicity is essential. Some studies have revealed a potential synergistic effect of natural compounds (NC) in combinatorial therapy with drugs, such as doxorubicin (Dox), which is frequently used for TNBC. Fucosterol (Fct), a phytosterol from brown seaweeds, seems a promising NC, because of its antioxidant ^[1,2] and antitumor effects ^[3,4]. The mechanisms underlying these effects are still controversial. 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 (using MTT/resazurin and BrdU assays, respectively), in monolayer and three-dimensional (3D) cultures. Additionally, we performed a histomorphological analysis of the 3D cultures. Data demonstrated that Fct (5 µM) alone did not affect cell viability and proliferation. In monolayer, Dox (1 µM) decreased less than 50% cell viability and proliferation, while in 3D this effect was only observed in cell viability using Dox 5 µM. The combination of Fct/Dox (5/0.1 µM) in monolayer, significantly decreased cell viability and proliferation, differing from the control group and from both compounds alone. These enhanching effects of Fct in monolayer were not observed in 3D, even using higher Dox concentrations. Morphometrical analyses revealed differences in 3D areas at Dox 5 µM. We conclude that Fct may increase Dox effects under certain conditions. One possible explanatory mechanism is that Fct may activate reactive oxygen species and trigger apoptosis via mitochondrial pathways ^[5]. We further conclude that the 3D cultures of TNBC were more resistant to treatments, corroborating other studies and reinforcing the use of these more complex models for drug screening.

Acknowledgements: To UIDB/04423/2020 and UIDP/04423/2020, provided by FCT and ERDF.

[1] DOI: 10.1007/BF02976680

[2] DOI: 10.1111/jphp.12404

[3] DOI 10.3233/BME-130876

[4] DOI: 10.4103/0973-1296.93327

[5] DOI:10.18632/oncotarget.13723

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 also 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 role. 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).

The continuous increase in daily exposure to ultraviolet radiation, which influence on 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 and healing compounds. One promising compound is the phytocannabinoid, cannabidiol (CBD) found in Cannabis sativa L., which has antioxidant and anti-inflammatory properties. Therefore, the main objective of this study was to compare the effect of CBD introduced into keratinocyte cultures in two ways: before and after UVB irradiation (pretreatment + treatment) and only after UVB irradiation (treatment) on the proteomic profile of the keratinocyte cell membrane proteins, which is the most exposed to environmental factors.

The data obtained from SDS-Page/nanoHPLC/QExactiveOrbiTrap showed that both UVB radiation and CBD treatments significantly induced changes in the proteomic profile of keratinocyte membranes. UVB has been shown to dramatically increase the expression of proteins involved in the regulation of translation/binding of RNA (40S S3a ribosomal protein), calcium ion homeostasis, differentiation, inflammatory response (S100-A6 and S100 proteins) and cellular redox status (Rho GDP 1 dissociation inhibitor). CBD when applied to cells 24h before and after UVB irradiation was found to reduce these UVB-induced changes more significant comparing to the cells treated with CBD only after irradiation. In contrast, treating keratinocytes only with CBD did not induce any significant changes for these proteins.

The results of this study showed that CBD significantly prevents UVB-induced changes in the proteomic profile of cell membranes. Moreover, the strong effect of CBD pretreatment suggests that this phytocannabinoid is meaningfully effective for protection of skin cells against the UVB-induced stress.

SA is supported by the project European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 754432.

Background: Chasmanthera dependens is a medicinal plant with wide application in African traditional medicine for the management of several pathologies. The phytochemistry, antiulcerogenic properties and antimicrobial properties have been reported and attributed to its robust phytochemistry. We hereby report the antioxidant properties of the chloroform extract of C. Dependens root as to provide further scientific information for the explanation of some of the reported properties of the plant in human diseases. Methods: In vitro studies assayed for diphpnylpicryl hydrazyl (DPPH) scavenging, ferric reducing antioxidant power (FRAP), hydrogen peroxide scavenging and total antioxidant capacity (TAC). Invivo studies assayed for superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) on CCl₄-induced wistar rats. Standard methods were followed in all assays. Results: DPPH radical scavenging activity was concentration dependent with an EC₅₀ of 647.67μg/ml. The extract showed a positive hydrogen peroxide scavenging with EC₅₀ of 57.78±2.93μg/ml relative to ascorbic acid standard (EC₅₀: 90.06μg/ml). The ferric reducing antioxidant power (FRAP) at the concentration 15.6μg/ml was 21.18±0.15 gallic acid equivalent while that of 1000μg/ml was 0.03±0.00 gallic acid equivalent for the extract. Total antioxidant capacity (TAC) at a concentration 15.6μg/ml showed a total antioxidant capacity of 15.22±7.81 ascorbic acid equivalent while that of 1000 μg/ml was 0.03±0.00 for the extract. Result of in vivo antioxidant studies showed a significant increase (p < 0.05) in the concentrations of endogenous antioxidant enzymes in the treated groups relative to the untreated control. Conclusion: we therefore conclude that the observed antioxidant properties of CECDR could be attributed to its wealth of phytochemical constituents as reported by previous studies. This study also highlights the possible application of CECDR as part of drug discovery raw material.