Hydrogen-rich Water Research Articles

The Anti-inflammatory Effects of Hydrogen-Rich Water Acts in a Volume-Dependent Manner on Rats’ Lungs Exposed to Cigarette Smoke

<em>Cigarette smoke can activate various ROS-sensitive signaling pathways in the lungs, triggering airway inflammation and leading to mucus hypersecretion. Meanwhile, hydrogen has shown anti-inflammatory activity in various injury models. This study aims to determine the anti-inflammatory effect of hydrogen-rich water on lungs exposed to cigarette smoke in rats. Male wistar rats were divided into four groups randomly (n=5), namely CI group (given aquades + exposed to free air), CII group (given aquades + exposed to cigarette smoke), HI group (given HRW 5 mL once a day + exposed to cigarette smoke), and HII group (given HRW 5 mL twice a day + exposed to cigarette smoke). HRW/aquades was administered orally 30 minutes before cigarette smoke exposure. Exposure to cigarette smoke lasts about 15-25 minutes with 5 cigarettes/day for 28 days. Subsequently, we examine the levels of lung NF-kB p65 using the ELISA method and perform pulmonary histopathological examination. The results showed that the administration of HRW reduced the levels of NF-κB p65 induced by cigarette smoke exposure, which is significant in the HII group (p<0.01). From the histopathological examination, administration of HRW significantly reduced the degree of lung inflammation caused by cigarette smoke exposure in the HI and HII group (p<0.05).</em>

The Effects of 12-Week Hydrogen-Rich Water Intake on Body Composition, Short-Chain Fatty Acids Turnover, and Brain Metabolism in Overweight Adults: A Randomized Controlled Trial

The aim of this randomized controlled trial was to analyze the effects of medium-term supplementation with hydrogen-rich water on brain metabolism, appetite-regulating hormones, body composition, and safety biomarkers in overweight adults. Twenty (n = 20, 10 females) apparently healthy adults with a body mass index >24.9 kg/m2 were assigned to receive 0.5 L per day of hydrogen-rich water (7.5 mg of hydrogen) or hydrogen-free water (tap water) for 12 weeks. Two-way analysis of variance with repeated measures revealed a significant difference between the two interventions in several body composition indices (P ≤ 0.05), with hydrogen-rich water superior to placebo to reduce waist circumference and mid-upper arm circumference by 1.31 cm (95% confidence interval, from –0.23 to 2.85) and 0.65 cm (95% confidence interval, from –0.10 to 1.40), respectively. Hydrogen-rich water outcompeted placebo to raise serum ghrelin levels, as the mean difference from the placebo group was 17.28 pmol/L (95% confidence interval, from 1.81 to 32.75) (P = 0.02). A non-significant strong positive trend (P = 0.10) was reported toward hydrogen-rich water being superior to placebo in augmenting total serum short-chain fatty acid levels, with a mean difference from the control group of 195.6 μmol/L (95% confidence interval, from –64.55 to 275.85). The mean fecal calprotectin levels were significantly reduced after hydrogen-rich water intervention for 19.7 μg/mg (95% confidence interval, from 0.31 to 39.09) (P = 0.03). Our findings advance hydrogen-rich water as a promising metabolic intervention in overweight adults, but further validation via multicentric longitudinal randomized controlled trials in metabolic and nutritional disorders is required.

P32-005-23 Effect of Ginseng Combined With Hydrogen-Rich Water Supplement on Blood Test of Dross-Country Skiers

P32-005-23 Effect of Ginseng Combined With Hydrogen-Rich Water Supplement on Blood Test of Dross-Country Skiers

Does hydrogen-rich water mitigate MP toxicity in rainbow trout (Oncorhyncus mykiss)? Monitoring with hematology, DNA damage, andapoptosis via ROS/GSH/MDA pathway

Although the number of studies documenting the presence of Microplastics (MP) in fish is increasing, research studies focused on its detoxification are very limited. In this study, rainbow trout (Oncorhyncus mykiss) were randomly divided into two groups after being fed with MPs (15% polypropylene [PP] +15% polyethylene [PE]) for 2 months. MP excretion without any application (PP+PE) in group I fish, and excretion of MPs with hydrogen-rich water (HRW) application (PP+PE+HRW) in group 2 were investigated under semi-static conditions for 21 days. This effectwas also compared by using positive and negative control groups (Control[no treatment, free PP, PE or/and HRW] and only HRW group). In this direction, the following were determined: PP+PE chronic toxicity in aquatic organisms, the toxicity mechanism and the effect of HRW as a possible treatment method in blood tissue; with hematological indices ([RBC count[RBC], leukocyte count [WBC], hemoglobin value [Hb], hematocrit ratio[Hct], platelet count [PLT], hemoglobin count per erythrocyte [MCHC], mean hemoglobin amount per erythrocyte [MCH] and mean erythrocyte volume [MCV]) in other tissues (liver, gill and brain tissue) oxidative stress response (catalase [CAT]), glutathione peroxidase (GPx), glutathionereductase (GR), superoxide dismutase (SOD), glutathione (GSH), reactiveoxygen products (ROS), malondialdehyde ([MDA] levels), DNA damage(8-OHdG: 8-Hydroxy-2-Deoxyguanosine), and the apoptosis (caspase 3)levels were investigated. In addition, acetylcholinesterase enzyme (AChE)activity, which is important in neurotoxicity pathways in the brain, was determined. The presence of plastics (PP/PE) in target tissues (muscle, liver, gill and gastrointestinal tract) was also obtained.The results showed that PP+PE caused toxicity in all three tissues. MPs showed an inhibiting effect on antioxidant enzyme activities and an inductive effect on MDA, ROS, 8-OHdG, and caspase 3 levels. HRW showed a mitigating effect on MP-mediated toxicity in O. mykiss brain, blood, gill, and liver by controlling the ROS/ GSH/MDA pathway. HRW can be suggested as a cost-effective and eco-friendly curative for the protection of fish from the oxidative damages produced by the ingestion of microplastics.

Effect and mechanism of hydrogen-rich bath on mice with imiquimod-induced psoriasis.

The purpose of this study was to investigate whether hydrogen-rich bath has therapeutic effect on psoriasis and its molecular mechanism. Mice with imiquimod-induced psoriasis were established and divided into groups. The mice were respectively treated with hydrogen-rich water bath and distilled water bath. The changes of skin lesions and PSI scores of mice were compared after their treatments. HE staining was used to observe the pathological feature. The changes of inflammatory indexes and immune factors were analysed by ELISA and immunohistochemical staining. Malondialdehyde (MDA) content was measured by the thiobarbituric assay (TBA) method. By naked eye, the severity of skin lesions in hydrogen-rich water bath group was lower than that in distilled water bath group, and the psoriasis severity index (PSI) was lower (p < 0.01). The results of HE staining showed that the mice with distilled water bath had more abnormal keratosis, thickening of the spinous layer and prolongation of the dermal process, and more Munro abscess than the mice with hydrogen-rich water bath. During the course of disease, the overall levels and peaks of IL-17, IL-23, TNF-α, CD3+ and MDA in mice with hydrogen-rich bath were lower than those in mice with distilled water bath (p < 0.05). In the skin, the mice treated with the hydrogen-rich water bath also had lower peak of proliferating cell nuclear antigen (PCNA) levels. It is concluded that hydrogen-rich water bath can inhibit psoriasis inflammation and oxidative stress, relieve psoriasis skin lesions and accelerate the end of abnormal skin proliferation state, which shows a therapeutic and improving effect on psoriasis.

Hydrogen-rich water treatment targets RT1-Db1 and RT1-Bb to alleviate premature ovarian failure in rats.

Premature ovarian failure (POF) is defined as the cessation of ovarian function before the age of 40 years, imposing a significant health burden on patients. However, effective etiological therapy for POF is scarce. Thus, we aimed to explore the protective role and targets of hydrogen-rich water (HRW) in POF. Based on cyclophosphamide (CTX)-induced POF rat models, the protective role of HRW treatment was mainly determined through serum 17-β-estradiol (E2), follicle-stimulating hormone (FSH), anti-mullerian hormone (AMH) levels, ovarian histomorphological analysis, and TUNEL assay. Tandem mass tag (TMT)-based quantitative proteomic analysis was then conducted on ovarian tissues, and the targets of HRW in POF were identified integrating differential expression analysis, functional enrichment analysis, and interaction analysis. In HRW treatment of POF rats, the serum AMH and E2 levels significantly increased, and FSH level significantly reduced, indicating the protective role of HRW. After TMT quantitative proteomic analysis, a total of 16 candidate differentially expressed proteins (DEPs) were identified after the cross analysis of DEPs from POF vs. control and POF+HRW vs. POF groups, which were found to be significantly enriched in 296 GO terms and 36 KEGG pathways. The crucial targets, RT1-Db1 and RT1-Bb, were finally identified based on both protein-protein interaction network and GeneMANIA network. The HRW treatment could significantly alleviate the ovarian injury of POF rats; RT1-Db1 and RT1-Bb are identified as two crucial targets of HRW treatment in POF rats.

Hydrogen-Rich Water Ameliorates Metabolic Disorder via Modifying Gut Microbiota in Impaired Fasting Glucose Patients: A Randomized Controlled Study.

Molecular hydrogen (H2) exhibits antioxidant, anti-inflammatory and anti-apoptotic effects, and has shown benefits in glucose and lipid metabolism in certain animal metabolic disorder models. However, the potential benefits of H2 treatment in individuals with impaired fasting glucose (IFG) has seldom been studied. This randomized controlled study (RCT) aims to investigate the effects of hydrogen-rich water (HRW) on IFG subjects and explore the underlying mechanism involved. Seventy-three patients with IFG were enrolled in a randomized, double-blind, placebo-controlled clinical study. These patients were assigned to receive either 1000 mL per day of HRW or placebo pure water (no H2 infusion) for a duration of eight weeks. Metabolic parameters and fecal gut microbiota were assessed at baseline (week 0) and at week 8. A combined analysis of metabolomics and intestinal microbiota was conducted to investigate the correlation between the effect of H2 on the metabolisms and the diversity of intestinal flora in the IGF patients. Both pure water and HRW demonstrated a significant reduction in fasting blood glucose in IFG patients, with a significant difference between pure water and HRW after eight weeks. Among IFG patients with abnormal pre-experimental fatty liver, 62.5% (10/16) in the HRW group and 31.6% (6/19) in the pure water group achieved remission. Furthermore, 16S RNA analysis revealed HRW-modified gut microbiota dysbiosis in the fecal samples of IGF patients. Through Pearson correlation analysis, the differential gut microbiota obtained by 16S analysis was found to be highly correlated with nine metabolites. H2 slightly improved metabolic abnormalities and gut microbiota dysbiosis, providing a novel target and theoretical basis for the prevention and treatment of blood glucose regulation in patients with IFG.

Hydrogen-rich water reduces cell damage by reducing excessive autophagy in mouse neuronal cells after oxygen glucose deprivation/reoxygenation

To investigate whether hydrogen-rich water exerts a protective effect against cellular injury by affecting the level of autophagy after oxygen glucose deprivation/reoxygenation (OGD/R) in a mouse hippocampal neuronal cell line (HT22 cells). HT22 cells in logarithmic growth phase were cultured in vitro. Cell viability was detected by cell counting kit-8 (CCK-8) assay to find the optimal concentration of Na2S2O4. HT22 cells were divided into control group (NC group), OGD/R group (sugar-free medium+10 mmol/L Na2S2O4 treated for 90 minutes and then changed to normal medium for 4 hours) and hydrogen-rich water treatment group (HW group, sugar-free medium+10 mmol/L Na2S2O4 treated for 90 minutes and then changed to medium containing hydrogen-rich water for 4 hours). The morphology of HT22 cells was observed by inverted microscopy; cell activity was detected by CCK-8 method; cell ultrastructure was observed by transmission electron microscopy; the expression of microtubule-associated protein 1 light chain 3 (LC3) and Beclin-1 was detected by immunofluorescence; the protein expression of LC3II/I and Beclin-1, markers of cellular autophagy, was detected by Western blotting. Inverted microscopy showed that compared with the NC group, the OGD/R group had poor cell status, swollen cytosol, visible cell lysis fragments and significantly lower cell activity [(49.1±2.7)% vs. (100.0±9.7)%, P < 0.01]; compared with the OGD/R group, the HW group had improved cell status and remarkably higher cell activity [(63.3±1.8)% vs. (49.1±2.7)%, P < 0.01]. Transmission electron microscopy showed that the neuronal nuclear membrane of cells in the OGD/R group was lysed and a higher number of autophagic lysosomes were visible compared with the NC group; compared with the OGD/R group, the neuronal damage of cells in the HW group was reduced and the number of autophagic lysosomes was notably decreased. The results of immunofluorescence assay showed that the expressions of LC3 and Beclin-1 were outstandingly enhanced in the OGD/R group compared with the NC group, and the expressions of LC3 and Beclin-1 were markedly weakened in the HW group compared with the OGD/R group. Western blotting assay showed that the expressions were prominently higher in both LC3II/I and Beclin-1 in the OGD/R group compared with the NC group (LC3II/I: 1.44±0.05 vs. 0.37±0.03, Beclin-1/β-actin: 1.00±0.02 vs. 0.64±0.01, both P < 0.01); compared with the OGD/R group, the protein expression of both LC3II/I and Beclin-1 in the HW group cells were notably lower (LC3II/I: 0.54±0.02 vs. 1.44±0.05, Beclin-1/β-actin: 0.83±0.07 vs. 1.00±0.02, both P < 0.01). Hydrogen-rich water has a significant protective effect on OGD/R-causing HT22 cell injury, and the mechanism may be related to the inhibition of autophagy.

Carbon dots-based reinforced hydrogen-rich water nanocomposite coating for storage quality of fresh-cut pear

Designing environmentally friendly edible nanocomposite films with antioxidant functions is of great interest in the field of fresh-cut fruits. Herein, Pleurotus eryngii carbon quantum dots (PER-CDs) were prepared from Pleurotus eryngii by-products, successfully characterized and added to chitosan (CS)/hydrogen-rich water solution to develop CDs/CS nanocomposite films (CS/PER-CDs), which was successfully applied to the preservation of fresh-cut pears. Thereinto, the fresh-cut pears treated with CS/2%PER-CDs nanocomposite films had minimum weight loss of 3.38% and the best retention of moisture, color, soluble solids and titratable acids after 5 days of storage. Importantly, the fresh-cut pears in the CS/2%PER-CDs treatment group had the lowest relative conductivity (37.53%) and MDA content of 1.94 μmol/kg, which could indicate that the CS/2%PER-CDs nanocomposite films maintained good cell membrane integrity of fresh-cut pears. In addition, the CS/2%PER-CDs treatment group effectively reduced the maximum respiration intensity (51.67 mgCO2/Kg·h) and ethylene production rate (0.75 μg/kg·h) of fresh-cut pear. Therefore, PER-CDs can be added into chitosan-based coating film as a functional component to make nanocomposite films and applied in fresh-cut fruits preservation effectively.

Hydrogen therapy as a potential therapeutic intervention in heart disease: from the past evidence to future application.

Cardiovascular disease is the leading cause of mortality worldwide. Excessive oxidative stress and inflammation play an important role in the development and progression of cardiovascular disease. Molecular hydrogen, a small colorless and odorless molecule, is considered harmless in daily life when its concentration is below 4% at room temperature. Owing to the small size of the hydrogen molecule, it can easily penetrate the cell membrane and can be metabolized without residue. Molecular hydrogen can be administered through inhalation, the drinking of hydrogen-rich water, injection with hydrogen-rich-saline, and bathing of an organ in a preservative solution. The utilization of molecular hydrogen has shown many benefits and can be effective for a wide range of purposes, from prevention to the treatment of diseases. It has been demonstrated that molecular hydrogen exerts antioxidant, anti-inflammatory, and antiapoptotic effects, leading to cardioprotective benefits. Nevertheless, the exact intracellular mechanisms of its action are still unclear. In this review, evidence of the potential benefits of hydrogen molecules obtained from in vitro, in vivo, and clinical investigations are comprehensively summarized and discussed with a focus on the cardiovascular aspects. The potential mechanisms involved in the protective effects of molecular hydrogen are also presented. These findings suggest that molecular hydrogen could be used as a novel treatment in various cardiovascular pathologies, including ischemic-reperfusion injury, cardiac injury from radiation, atherosclerosis, chemotherapy-induced cardiotoxicity, and cardiac hypertrophy.

Highly Active Hydrogen-rich Photothermal Reverse Water Gas Shift Reaction on Ni/LaInO3 Perovskite Catalysts with Near-unity Selectivity.

Photo-assisted reverse water gas shift (RWGS) reaction is regarded green and promising in controlling the reaction gas ratio in Fischer Tropsch synthesis. But it is inclined to produce more byproducts in high H2 concentration condition. Herein, LaInO3 loaded with Ni-nanoparticles (Ni NPs) was designed to obtain an efficient photothermal RWGS reaction rate, where LaInO3 was enriched with oxygen vacancies to roundly adsorbing CO2 and the strong interaction with Ni NPs endowed the catalysts with powerful H2 activity. The optimized catalyst performed a large CO yield rate (1314 mmol gNi -1 h-1 ) and ≈100 % selectivity. In situ characterizations demonstrated a COOH* pathway of the reaction and photoinduced charge transfer process for reducing the RWGS reaction active energy. Our work provides valuable insights on the construction of catalysts concerning products selectivity and photoelectronic activating mechanism on CO2 hydrogenation.

The Effects of Medium-Term Intake of Hydrogen-Rich Water on Sperm Quality Biomarkers in Normospermic and Oligospermic Men: A Randomized Controlled Pilot Trial

The main aim of this randomized controlled pilot trial was to evaluate the effects of hydrogen-rich water on spermiogram parameters in normospermic and oligospermic men. As many as 12 healthy young men (age 29.1 ± 5.9 years; n = 6 normospermic; n = 6 oligospermic) volunteered. Participants were allocated in a double-blind manner to receive 1 L of hydrogen-rich water per day or 1 L of tap water fortified with magnesium for 8 weeks. Following hydrogen-rich water supplementation, sperm concentration and morphology tended to increase by 12.4 million per milliliter (95% CI; –31.8 to 56.6), and live cells increased by 3.8% (95% CI; –12.5 to 20.1), respectively (P ≤ 0.30). A significant difference between the two interventions was found in sperm vitality (P = 0.03), with hydrogen-rich water being superior to the placebo in terms of an increase in the number of live sperm cells. Subgroup analyses revealed no significant differences between interventions in sperm viability outcomes in oligospermic men (P &gt; 0.05). However, sperm count increased by over 20 million in two of the three oligospermic men (66.7%) receiving hydrogen-rich water, while no oligospermic men in the placebo group increased sperm concentration to euspermic levels (P &lt; 0.01). These preliminary findings suggest hydrogen-rich water is an effective intervention for improving some aspects of male subfertility; further large-sample trials are warranted to corroborate our results.

Effect of Hydrogen-Enriched Solvents on the Extraction of Phytochemicals in Propolis.

Propolis, one of the most important bee products, cannot be used in its raw form. The efficiency of the bioactive components of propolis increases with the extraction process. The choice of solvent to be used in the extraction of propolis is effective in determining the properties of the extract. Ethanol is the most widely used solvent, which significantly increases the efficiency of its bioactive components in the extraction of propolis. Effective nonalcohol-based extraction techniques have become important since alcohol-based extracts cause some discomfort and cannot be used in people with alcohol intolerance. The use of water in propolis extraction is less preferred than ethanol because it does not thoroughly dissolve the bioactive components. In this study, the effect of incorporating hydrogen into solvents (water, ethanol, and methanol) on the extraction of total phenolic content, total flavonoid content, antioxidant activities, and phenolic compound profile of the propolis sample was evaluated. Incorporation of H2 into water, ethanol, and methanol led to an increase in total phenolic content by 19.08, 5.43, and 12.71% and in the total flavonoid content by 28.97, 17.13, and 2.06%, respectively. Besides, the highest increases in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) scavenging activities were observed in hydrogen-rich water (4.4%) and hydrogen-rich ethanol (32.4%) compared to their counterparts, respectively. On the other hand, incorporation of H2 into different solvents led to significant increases in different phenolics, and it was observed that the level of change was dependent on the type of the phenolic compound and the solvent used. This study is important in terms of using hydrogen-enriched solvents to extract phenolics from propolis for the first time. Using hydrogen-rich solvents, specifically hydrogen-rich water, was observed to be an effective method for the improvement of phytochemical extraction efficiency in propolis.

The Role and Mechanism of Hydrogen-Rich Water in the Cucumis sativus Response to Chilling Stress

Cucumber is a warm climate vegetable that is sensitive to chilling reactions. Chilling can occur at any period of cucumber growth and development and seriously affects the yield and quality of cucumber. Hydrogen (H2) is a type of antioxidant that plays a critical role in plant development and the response to stress. Hydrogen-rich water (HRW) is the main way to use exogenous hydrogen. This study explored the role and mechanism of HRW in the cucumber defense response to chilling stress. The research results showed that applying 50% saturated HRW to the roots of cucumber seedlings relieved the damage caused by chilling stress. The growth and development indicators, such as plant height, stem diameter, leaf area, dry weight, fresh weight, and root length, increased under the HRW treatment. Photosynthetic efficiency, chlorophyll content, and Fv/Fm also improved and reduced energy dissipation. In addition, after HRW treatment, the REC and MDA content were decreased, and membrane lipid damage was reduced. NBT and DAB staining results showed that the color was lighter, and the area was smaller under HRW treatment. Additionally, the contents of O2- and H2O2 also decreased. Under chilling stress, the application of HRW increased the activity of the antioxidases SOD, CAT, POD, GR, and APX and improved the expression of the SOD, CAT, POD, GR, and APX antioxidase genes. The GSSG content was reduced, and the GSH content was increased. In addition, the ASA content also increased. Therefore, exogenous HRW is an effective measure for cucumber to respond to chilling stress.

Short-term intake of hydrogen-rich water positively affects neuropsychological performance in young adults

Short-term intake of hydrogen-rich water positively affects neuropsychological performance in young adults

Hydrogen-rich water as a green solvent for the extraction of phytochemicals from agri-food wastes

The valorization of agri-food waste forms a pressing need in the present crop unavailability crisis. Phytochemical extraction may provide an attractive solution to this problem. The use of hydrogen-rich water (HRW) as a solvent for extracting different phytochemicals from various wastes was investigated. The total phenolic content (TPC), total flavonoids content (TFC), total anthocyanins content, and antioxidant activity (DPPH, ABTS+) of HRW extracts were higher than those of pure water, and those of ethanol/HRW (50/50) were higher than those of ethanol/water (50/50) for all waste types. The highest levels of all phytochemicals were found in HRW extracts, followed by ethanol/HRW (50/50). The highest phytochemical contents were shown in HRW samples for all plant wastes as follows: TPC (mg GAE/g extract): 9.04 (tomato peels), 9.32 (carrot), 14.76 (apple peels), 23.88 (lemon peels), and 49.15 (cabbage); TFC, (mg QE/g extract): 9.37 (tomato peels), 10.84 (carrot), 8.96 (apple peels), 9.37 (lemon peels), and 13.31 (cabbage); total anthocyanins (mg QE/g extract): 2.88 (tomato peels), 6.14 (carrot), 4.01 (apple peels), 1.63 (lemon peels), and 40.73 (cabbage); DPPH scavenging activity (mg AE/g extract): 8.47 (tomato peels), 11.06 (carrot), 8.40 (apple peels), 9.85 (lemon peels), and 20.91 (cabbage); and ABTS + scavenging activity (mg TE/g extract): 8.61 (tomato peels), 8.36 (carrot), 11.69 (apple peels), 11.87 (lemon peels), and 24.25 (cabbage). The increase rate was 22–50% for TPC, 16-53% for TFC, 80-390% for total anthocyanins, 9–142% for DPPH scavenging activity, and 14–28% for ABTS+ scavenging activity when HRW was used as a solvent instead of pure water. The rate of increase was 1-23% for TPC, 53-45% for TFC, 36-228% for total anthocyanins, 19-171% for DPPH scavenging activity, and 5-30% for ABTS+ scavenging activity when HRW, mixed with equal amounts of ethanol (50/50), was used as a solvent instead of pure water/ethanol (50/50). The quantities of certain flavonoids and non-flavonoids such as epicatechin, rutin, gallic acid, and p-coumaric acid increased at rates reaching several folds when HRW was used as the solvent. The results of extract color were in line with those of chemical ones, where the highest C* values were observed for HRW, followed by ethanol/HRW (50/50). A positive linear correlation (R2 = 0.61–0.94) between several color parameters (H°, C*, a*) and anthocyanin content was observed. The results showed that using HRW alone or mixed with ethanol significantly improves the extraction of different phytochemicals. The brilliant advantage of this method is that it does not need additional equipment, energy, and toxic solvents, allowing it to be a low-cost and environmentally friendly method.

Use of hydrogen-rich water as a green solvent for the extraction of phytochemicals: Case of olive leaves

Olive leaves are a rich source of phytochemicals such as phenolic compounds. ‎Extraction of phytochemicals is the best strategy for valorizing ‎by-products. Any extraction method is chosen based on its economic feasibility, safety, and environmental friendliness. In the present study, the extraction of phenolic compounds (TPC), flavonoids (TFC), and antioxidants (DPPH and ABTS) from the olive leaf was assayed using two types of hydrogen-rich water: hydrogen bubbling (HRW) and magnesium incorporation (Mg water)‎. The highest extraction yield was found for HRW at 34.42% and the lowest one was for pure water at 19.27%. The highest values of TPC, TFC, and antioxidant activity (DPPH and ABTS) were shown for HRW water ‎followed by Mg water/ethanol (50/50) samples, whereas the lowest ones were observed for pure water samples‎. ‎The incorporation of hydrogen into water increased TPC, TFC, DPPH, and ABTS by ‎‎95.63, ‎290.62, ‎‎60.78‎, and ‎118.94%, respectively; whereas the incorporation of Mg increased them by ‎‎7.55, ‎16.49, ‎‎14.23, and ‎8.35%, respectively.‎ Trans-ferulic acid, rosmarinic acid, p-‎coumaric acid, and rutin increased by 1.3, 5.5, 20, and 23 folds when hydrogen was ‎incorporated ‎into water, whereas catechin appeared only ‎in HRW.‎ HRW can be suggested as a green and eco-friendly solvent for improving the extraction of phytochemicals.

Hydrogen-rich water treatment increased several phytohormones and prolonged the shelf life in postharvest okras.

Hydrogen-rich water (HRW) treatment has been reported to delay the softening and senescence of postharvest okras, but its regulatory mechanism remains unclear. In this paper, we investigated the effects of HRW treatment on the metabolism of several phytohormones in postharvest okras, which act as regulatory molecules in fruit ripening and senescence processes. The results showed that HRW treatment delayed okra senescence and maintained fruit quality during storage. The treatment upregulated all of the melatonin biosynthetic genes such as AeTDC, AeSNAT, AeCOMT and AeT5H, contributing to the higher melatonin content in the treated okras. Meanwhile, increased transcripts of anabolic genes but lower expression of catabolic genes involved in indoleacetic acid (IAA) and gibberellin (GA) metabolism were observed in okras when treated with HRW, which was related to the enhanced levels of IAA and GA. However, the treated okras experienced lower abscisic acid (ABA) content as compared to the non-treated fruit due to the down-regulation of its biosynthetic genes and up-regulation of the degradative gene AeCYP707A. Additionally, there was no difference in γ-aminobutyric acid between the non-treated and HRW-treated okras. Collectively, our results indicated that HRW treatment increased levels of melatonin, GA and IAA, but decreased ABA content, which ultimately delayed fruit senescence and prolonged shelf life in postharvest okras.

Molecular Hydrogen Prevents Osteoclast Activation in a Glucocorticoid-Induced Osteoporosis Zebrafish Scale Model.

Antioxidants represent a powerful tool for many human diseases and, in particular, molecular hydrogen has unique characteristics that make it a very promising therapeutic agent against osteoporosis. Zebrafish scales offer an innovative model in which new therapeutic approaches against secondary osteoporosis are tested. Scale bone loss obtained by prednisolone (PN) treatment is characterized by increased osteoclast activity and decreased osteoblast activity highlighted with bone enzymatic assays. We used this read-out system to test the therapeutic effects of hydrogen-rich water (HRW), an innovative antioxidant approach. HRW prevented osteoclast activation and bone loss in PN-treated fish scales, as verified by both biochemical and histochemical tartrate-resistant alkaline phosphatase assays. On the other hand, HRW treatment did not prevent PN-dependent osteoblast suppression, as measured by alkaline phosphatase activity. Moreover, HRW treatment did not facilitate the reparation of resorption lacunae induced in scales by PN. Our study highlighted a specific effect of HRW on adult osteoclast activity but not in osteoblasts, introducing an intriguing new antioxidant preventive approach against osteoporosis.

Comparative study of phytochemical extraction using hydrogen-rich water and supercritical fluid extraction methods

Phytochemicals are widely found in agri-food wastes. The extraction of ‎phytochemicals was evaluated using hydrogen-rich water extraction (HRW-E) and supercritical fluid ‎extraction (SFE) methods from different plant wastes (tomato peel, orange carrot, green ‎apple ‎peel, lemon peel, red cabbage). Total phenolic content (TPC), total ‎flavonoid content (TFC), total anthocyanin, and antioxidant activity (DPPH and ABTS) were ‎highest in HRW-E samples compared to SFE and pure water extraction for all plant wastes. The percent increase (%) ranged between 5.14 and 12.06 and 22.27–49.62 (TPC), 1.32–35.59 and ‎‎16.01-‎‎‎53.03 (TFC), 18.18–53.19 and 80.53–390.1 (anthocyanins), 2.21–22.37 and 9.03-‎‎142.46 ‎‎‎(DPPH), and 1.16–7.49 and 14.47–28.05 (ABTS) for SFE and HRW-E samples, ‎respectively. ‎HRW-E was more potent than SFE for extraction of all phytochemical types from ‎all plant wastes. Non-flavonoids, e.g., phenolic acids (gallic acid, chlorogenic acid, p-‎coumaric acid, trans-ferulic acid‎) and flavonoids (catechin, epicatechin, and rutin)‎, were extracted ‎better by HRW-E than ‎‎SFE for different plant wastes. ‎The HRW-E method is recommended to researchers and processors‎ as an extraction method for phytochemicals due to its simplicity, low cost, not requiring additional equipment or ‎energy source, non-toxic solvent use, green properties of the product and for the environment, as well as higher extraction ‎‎efficiency.‎