Reductase Gene Expression Research Articles

Thermal preconditioning in a reef-building coral alleviates oxidative damage through a BI-1-mediated antioxidant response

Global coral reef decline is largely driven by the breakdown of the coral-algal symbiosis during temperature stress. Corals can acclimatize to higher temperatures, but the cellular processes underlying this ability are poorly understood. We show that preconditioning-based improvements in thermal tolerance in Pocillopora acuta are accompanied by increases in host glutathione reductase (GR) activity and gene expression, which prevents DNA damage. A strong correlation between GR and BI-1 expressions in heat-stressed preconditioned corals and the presence of an antioxidant response element (ARE) in the GR promoter suggest BI-1 could regulate GR expression through Nrf2/ARE pathway. To fortify this link, we developed and GFP-validated an siRNA-mediated gene knockdown protocol and targeted the coral BI-1 gene. BI-1 knock-down specifically decreased GR expression and activity and increased oxidative DNA damage in heat-stressed preconditioned corals, showing that a BI-1-mediated, enhanced antioxidant response during acute heat stress is a key mechanism that prevents oxidative DNA damage after preconditioning.

Limited Zn supply affects nutrient distribution, carbon metabolism and causes nitro-oxidative stress in sensitive Brassica napus

Despite being a worldwide problem, responses of crops to zinc (Zn) deficiency is largely unknown. This study examines the effects of limited Zn supply in Brassica napus . The decreased Zn content of the nutrient solution caused reduced tissue Zn concentrations and reduced K, Ca, Mg levels were detected in the leaves. For Ca, Zn, Fe, Mo inhomogeneous distribution was induced by Zn limitation. Suboptimal Zn supply decreased the shoot and root biomass and altered the root structure. The levels of sugars and sugar phosphates (e.g. glucose, glucose-6-phosphate etc.) were decreased suggesting disturbance in carbon metabolism. Limited Zn availability induced organ-dependent superoxide anion and hydrogen peroxide production and triggered altered NADPH oxidase, superoxide dismutase activities, ascorbate peroxidase abundance and ascorbate and glutathione contents indicating oxidative stress. Moreover, nitric oxide, peroxynitrite and S-nitrosoglutathione (GSNO) levels and GSNO reductase gene expression, protein level and activity were modified by Zn limitation. As a consequence, protein tyrosine nitration and protein carbonylation were intensified in B. napus grown with suboptimal Zn supply. Collectively, these results provide the first evidence for Zn deficiency-induced imbalances in nutrient status, sugar contents, reactive oxygen and nitrogen species metabolisms and for the secondary nitro-oxidative stress in sensitive Brassica napus . • Brassica napus is sensitive to Zn limitation. • Zn deficiency is accompanied by disturbed nutrient homeostasis in Brassica leaves. • Zn limitation disturbs carbon metabolism indicated by reduced sugar contents. • Zn deficiency induces oxidative stress due to disturbed antioxidant defence. • Zn limitation disturbs reactive nitrogen species levels leading to nitro-oxidative stress.

Caffeic acid regulated respiration, ethylene and reactive oxygen species metabolism to suppress senescence of Malus domestica

The current study assessed the variations of reactive oxygen species (ROS), respiration and ethylene metabolism in ‘Qiujin’ apples after caffeic acid dipping. Results demonstrated that caffeic acid remarkably inhibited respiratory rate, ethylene release rate and hydrogen peroxide accumulation, but increased reduced glutathione and ascorbic acid contents in apples. Caffeic acid also improved catalase, peroxidase, superoxide dismutase, nicotinamide adenine dinucleotide phosphate oxidase, dehydroascorbate reductase, monodehydroascorbate reductase, glutathione reductase and ascorbate peroxidase activities and gene expressions in apples. In addition, caffeic acid up-regulated MdG6PDH6 and MdAOX expressions, down-regulated MdACS1, MdACO2, MdPK, MdHK3, MdPFK, MdSDH and MdCCO expressions in the fruit. Taken together, caffeic acid could regulate the key enzymes activities and gene expressions in respiration, ethylene and ROS metabolism to suppress senescence of apples after harvest.

From Photocatalysis to Photo-Electrocatalysis: An Innovative Water Remediation System for Sustainable Fish Farming

In this study, the effects of photo-electrocatalysis (PEC) were evaluated as an innovative application of conventional photocatalysis (PC) to remediate water in a recirculating system for rainbow trout (Oncorhynchus mykiss) culture, in relation to fish welfare and health, with a multidisciplinary approach. Three tanks were employed, equipped with conventional biological filters as a control system, and three tanks equipped with the PEC purification system. The concentrations of ammonia, nitrite and nitrate ions in water were monitored, and the fish’s oxidative damage and stress response were evaluated in parallel. The water of the PEC-treated experimental group showed lower ammonia (TAN) and nitrite concentrations and higher nitrate concentration, possibly deriving from TAN oxidation through PEC, also leading to gaseous N2. Histological analysis did not reveal any pathological alteration in the gills and liver of both groups. The superoxide dismutase (sod1), glutathione reductase (GR), glutathione peroxidase (GPx1), and Tumor necrosis factor (TNFα) gene expressions were significantly higher in the control group than in the PEC-treated group, while the Heat shock protein 70 (Hsp70) expression did not show any difference in the two groups. These results indicate that the use of PEC filters has a positive effect on water quality, compared to the use of conventional biological filters, inducing a high level of welfare in O. mykiss.

Essential Roles of Ribonucleotide Reductases under DNA Damage and Replication Stresses in Cryptococcus neoformans.

ABSTRACTA balance in the deoxyribonucleotide (dNTPs) intracellular concentration is critical for the DNA replication and repair processes. In the model yeast Saccharomyces cerevisiae, the Mec1-Rad53-Dun1 kinase cascade mainly regulates the ribonucleotide reductase (RNR) gene expression during DNA replication and DNA damage stress. However, the RNR regulatory mechanisms in basidiomycete fungi during DNA replication and damage stress remain elusive. Here, we observed that in C. neoformans, RNR1 (large RNR subunit) and RNR21 (one small RNR subunit) were required for cell viability, but not RNR22 (another small RNR subunit). RNR22 overexpression compensated for the lethality of RNR21 suppression. In contrast to the regulatory mechanisms of RNRs in S. cerevisiae, Rad53 and Chk1 kinases cooperatively or divergently controlled RNR1 and RNR21 expression under DNA damage and DNA replication stress. In particular, this study revealed that Chk1 mainly regulated RNR1 expression during DNA replication stress, whereas Rad53, rather than Chk1, played a significant role in controlling the expression of RNR21 during DNA damage stress. Furthermore, the expression of RNR22, not but RNR1 and RNR21, was suppressed by the Ssn6-Tup1 complex during DNA replication stress. Notably, we observed that RNR1 expression was mainly regulated by Mbs1, whereas RNR21 expression was cooperatively controlled by Mbs1 and Bdr1 as downstream factors of Rad53 and Chk1 during DNA replication and damage stress. Collectively, the regulation of RNRs in C. neoformans has both evolutionarily conserved and divergent features in DNA replication and DNA damage stress, compared with other yeasts.IMPORTANCE Upon DNA replication or damage stresses, it is critical to provide proper levels of deoxynucleotide triphosphates (dNTPs) and activate DNA repair machinery. Ribonucleotide reductases (RNRs), which are composed of large and small subunits, are required for synthesizing dNTP. An imbalance in the intracellular concentration of dNTPs caused by the perturbation of RNR results in a reduction in DNA repair fidelity. Despite the importance of their roles, functions and regulations of RNR have not been elucidated in the basidiomycete fungi. In this study, we found that the roles of RNR1, RNR21, and RNR22 genes encoding RNR subunits in the viability of C. neoformans. Furthermore, their expression levels are divergently regulated by the Rad53-Chk1 pathway and the Ssn6-Tup1 complex in response to DNA replication and damage stresses. Therefore, this study provides insight into the regulatory mechanisms of RNR genes to DNA replication and damage stresses in basidiomycete fungi.

Cornelian cherry (Cornus mas L.) fruit extract improves growth performance, disease resistance, and serum immune-and antioxidant-related gene expression of common carp (Cyprinus carpio)

This study evaluated the use of cornelian cherry (Cornus mas L.) fruit extract on the growth performance, immune responses, and disease resistance in common carp (Cyprinus carpio). A total of 192 fish (initial body weight of 18.34 ± 1.8 g) were randomly distributed in 12 tanks with 16 fish per tank (4 treatments with 3 replications) and fed with diets containing cornelian cherry extract (CCE11CCE (cornelian cherry extract)) at the rate of 0, 0.25, 0.5 and 1%. The final weight of common carp fed CCE at 0.5% was significantly higher than fish fed 0 and 0.25% doses (p < 0.05). In addition, weight gain and specific growth rate were increased, and feed conversion ratio decreased in fish-fed diets containing 0.5 and 1% CCE compared to other groups. The challenge test with A. hydrophila revealed that common carp-fed CCE had a lower mortality rate than the control. The lysozyme activity was markedly higher in common carp fed 0.5% of cornelian cherry extract than in control. The total immunoglobulin and total protein showed higher values in fish fed 0.5 and 1% than the control. The catalase gene showed higher mRNA levels in fish fed 0.5 and 1% than the control. Glutathione reductase gene expression was markedly upregulated in common carp fed 1% of CCE than the control and 0.25% groups. The glutathione S-transferase gene showed higher mRNA levels in fish fed 0.5 and 1% than the control and 0.25% groups. The superoxide dismutase gene expression was markedly higher in common carp fed 0.5% of CCE than in control. The CC-chemokine and interleukin-10 showed higher mRNA levels in fish fed 0.5% than the remaining groups. Hypoxia-inducible factor 1-alpha gene expression was markedly upregulated in common carp fed 1% of CCE than the control and 0.25% groups. The heat shock 70 kDa protein and interleukin-1 beta genes showed higher mRNA levels in fish fed 0.25, 0.5, and 1% than the control. In conclusion, the supplementation of CCE at 0.5–1% is recommended to enhance the growth performance and relieve the impacts of A. hydrophila infection in common carp.

Effect ofmelatonin onthe gastric antioxidant defence inexperimental burn trauma.

Severe burn trauma triggers oxidative gastric mucosal injury. The purpose ofthis study was toinvestigate the antioxidant defence mechanisms and protective effect ofmelatonin inthe gastric mucosa after burn injury. Inorder toinvestigate the mechanisms involved inthe gastric antioxidant defence inarat burn model, quantitative real-time PCR and immunohistochemistry techniques were applied. Ananalysis ofglutathione peroxidase4 (GPx4), glutathione reductase (GR), and catalase (Cat) gene expression was performed along with the evaluation ofthe gastric Cu/Zn superoxide dismutase (Cu/Zn SOD) activity. Melatonin was applied immediately and 12h after 30% oftotal body surface area burns. The burn injury significantly increased the Gpx4 mRNA (P<0.0001) and Gsr mRNA (P<0.0001) expression. Italso had aslight positive effect onthe Cat mRNA expression and Cu/Zn SOD activity. Melatonin, inturn, markedly augmented the burn-induced Cu/Zn SOD (P<0.0001) activity, reversed the Gpx4 mRNA (P<0.0001) and Gsr mRNA (P<0.0001) expression, and inhibited the Cat mRNA level. Inconclusion, the present study suggests that aburn injury adaptively increases the Cu/Zn SOD activity and enhances the Gpx4 and Gsr gene expression inthe gastric mucosa. Melatonin effectively modulates the expression ofthe cellular antioxidant enzymes, and improves the antioxidant defence byaugmenting the Cu/Zn SOD activity.

Nitric oxide (NO) involved in Cd tolerance in NHX1 transgenic duckweed during Cd stress

ABSTRACT Anthropogenic activities cause heavy metal pollution, such as cadmium (Cd). Na+/H+ antiporter (NHX1) transgenic duckweed showed Cd tolerance in our previous study, and the signal mechanism needs to be explored. As an important signal molecule, nitric oxide (NO) is involved in a number of functions under abiotic stress response. This study analyzed the levels of endogenous NO in wild-type (WT) duckweed and NHX1 duckweed under Cd treatment. The results showed that after 24 h Cd treatment, the endogenous NO level of WT duckweed decreased, which was significantly lower than that in NHX1 duckweed. Studies have proved that NHX1 influences pH. The level of NO in this study has been investigated at different pH. The NO level was the highest in the duckweed cultured with pH 5.3. Nitrate reductase gene expression was down-regulated and NO synthesis was decreased under Cd stress in WT duckweed. This study showed that NO level has been modified in NHX1 duckweed, which could be influcened by pH.

Metabolic Pathways Involved in the Drought Stress Response of Nitraria tangutorum as Revealed by Transcriptome Analysis

Drought resistance in plants is controlled by multiple genes. To identify the genes that mediate drought stress responses and to assess the associated metabolic pathways in the desert shrub Nitraria tangutorum, we conducted a transcriptome analysis of plants under control (maximum field capacity) and drought (20% of the maximum field capacity) conditions. We analyzed differentially expressed genes (DEGs) of N. tangutorum and their enrichment in the KEGG metabolic pathways database, and explored the molecular biological mechanisms underlying the answer to its drought tolerance. Between the control and drought groups, 119 classified metabolic pathways annotated 3047 DEGs in the KEGG database. For drought tolerance, nitrate reductase (NR) gene expression was downregulated, indicating that NR activity was decreased to improve drought tolerance. In ammonium assimilation, drought stress inhibited glutamine formation. Protochlorophyllide reductase (1.3.1.33) expression was upregulated to promote chlorophyll a synthesis, whereas divinyl reductase (1.3.1.75) expression was downregulated to inhibit chlorophyll-ester a synthesis. The expression of the chlorophyll synthase (2.5.1.62) gene was downregulated, which affected the synthesis of chlorophyll a and b. Overall, drought stress appeared to improve the ability to convert chlorophyll b into chlorophyll a. Our data serve as a theoretical foundation for further elucidating the growth regulatory mechanism of desert xerophytes, thereby facilitating the development and cultivation of new, drought-resistant genotypes for the purpose of improving desert ecosystems.

Compost and microbial biostimulant applications improve plant growth and soil biological fertility of a grass-based phytostabilization system

In this work, a grass-based phytoremediation system integrated with an organic amendment and biostimulants was evaluated for remediating contaminated sites. Plant growth and biological fertility were monitored to assess the efficacy of a vegetative cap used as a safety measure to reduce sanitary and environmental risks of industrially contaminated soils and soil-washing sludges. Both matrices were potentially contaminated with Pb and Zn with an ecological risk index from low to moderate. According to potentially toxic elements (PTEs) bioaccessibility tests, the exposure to the released fine particulate matter may cause serious risks to human beings, in particular to children. The grass mixture was well adapted to both the substrates and a low PTEs mobility was detected, thus, reducing the leaching risk to ground water sources. Compost addition augmented significantly nitrogenase reductase (nifH) and ammonia monooxygenase (amoA) gene expression abundance in both substrates. Furthermore, a positive interaction between compost fertilization and a Trichoderma-based biostimulant inoculation was recorded in sludges resulting in a significant stimulation of nitrogen-fixing and ammonia-oxidizing bacteria. The application of compost and biostimulant increased soil fertility and plant growth. Furthermore, there was a slight reduction in PTE bioaccessibility, thus, improving the efficiency of the phytostabilization, limiting the resuspension and dispersion of the health-risk soil particulate.

Enhanced resistance in ‘shatang’ mandarin fruit against Penicillium italicum caused by 2-methoxy-1,4-naphthoquinone

Blue mold caused by Penicillium italicum leads to considerable postharvest losses in citrus fruit. The 2-methoxy-1,4-naphthoquinone (MNQ), a natural compound from Chinese medicinal plant, had strong in vitro antifungal activity. Therefore, we investigated the in vivo effect of MNQ treatment against blue mold on ‘shatang’ mandarin fruit caused by P. italicum and the underlying mechanisms. Our results found that MNQ could suppress disease development, indicated by less severe incidence and symptoms of fruit disease. MNQ treatment maintained redox status via enhancing antioxidant enzyme activities to decrease hydrogen peroxide (H2O2) content and up-regulating sulfoxide reductase, glutaredoxin and thioredoxin gene expression to keep protein repair ability. Meanwhile, MNQ alleviated membrane damage and protein carbonylation caused by oxidative stress. Additionally, MNQ increased the activities of defense-related enzymes (phenylalanine ammonia lyase, β-1,3-glucanase and chitinase) and accumulation of lignin, phenolic compounds and flavonoids. Moreover, we found that MNQ treatment could up-regulate abscisic acid (ABA) synthetic gene expression resulting in higher ABA content. In summary, MNQ was effective as a potential agent controlling the postharvest diseases.

In Vitro and In Vivo Regulation of SRD5A mRNA Expression of Supercritical Carbon Dioxide Extract from Asparagus racemosus Willd. Root as Anti-Sebum and Pore-Minimizing Active Ingredients.

Oily skin from overactive sebaceous glands affects self-confidence and personality. There is report of an association between steroid 5-alpha reductase gene (SRD5A) expression and facial sebum production. There is no study of the effect of Asparagus racemosus Willd. root extract on the regulation of SRD5A mRNA expression and anti-sebum efficacy. This study extracted A. racemosus using the supercritical carbon dioxide fluid technique with ethanol and investigated its biological compounds and activities. The A. racemosus root extract had a high content of polyphenolic compounds, including quercetin, naringenin, and p-coumaric acid, and DPPH scavenging activity comparable to that of the standard L-ascorbic acid. A. racemosus root extract showed not only a significant reduction in SRD5A1 and SRD5A2 mRNA expression by about 45.45% and 90.86%, respectively, but also a reduction in the in vivo anti-sebum efficacy in male volunteers, with significantly superior percentage changes in facial sebum production and a reduction in the percentages of pore area after 15 and 30 days of treatment. It can be concluded that A. racemosus root extract with a high content of polyphenol compounds, great antioxidant effects, promising downregulation of SRD5A1 and SRD5A2, and predominant facial sebum reduction and pore-minimizing efficacy could be a candidate for an anti-sebum and pore-minimizing active ingredient to serve in functional cosmetic applications.

Yellow Yeast Rice Prepared Using Aspergillus terreus DSMK01 Lowers Cholesterol Levels by Stimulating Bile Salt Export Pump in Subjects with Mild-to-Moderate Hypercholesterolemia: A Randomized Controlled Trial.

Aspergillus terreus is an industrial microorganism used in the brewing and sauce industries. It produces monacolin K, a natural statin. The study conducted an 8-week randomized controlled trial with hypercholesterolemic subjects to examine the hypocholesterolemic effects and mechanisms of supplementation with yellow yeast rice (YYR) prepared by growing Aspergillus fungi on steamed rice. YYR supplementation markedly reduced total cholesterol, LDL, and apolipoprotein B100 levels in plasma compared with the placebo. In addition, YYR induced a significantly increased ATP binding cassette subfamily B member 11 (ABCB11) gene expression compared with the placebo, indicating the role of YYR in lowering intrahepatic cholesterol availability by stimulating the bile salt export pump. Upregulation of LDL receptor (LDLR) and 3-methylglutaryl-CoA reductase (HMGCR) gene expressions provided additional evidence to support the role of YYR in reducing hepatic cholesterol availability. Plasma metabolomic profiling revealed the possibility of diminishing bile acid absorption. Finally, Spearman rank analysis showed correlations of plasma cholesterol profiles with HMGCR and LDLR gene expressions (negative) and plasma bile acids (positive). Plasma bile acids also correlated with ABCB11 (negative) and LDLR (positive) gene expressions. These findings suggest that daily YYR supplementation exerted hypocholesterolemic effects in mild-to-moderate hypercholesterolemic subjects by reducing intrahepatic cholesterol availability through stimulating bile salt export pumps and inhibiting cholesterol biosynthesis.

Novel vitamin D3-hydroxyderivatives as candidates for the therapy against skin-aging and photo-aging: bioinformatical analysis

Vitamin D3 acts through its most active form, calcitriol, 1α,25-dihydroxyvitamin D3 [1,25(OH)2D3] as agonist of one of the receptors involved in this ligand action, vitamin D receptor (VDR), which is also a transcription factor. Numerous modifications of calcitriol at its side-chain, C-ring, A-ring, triene system, alone or in combination, as well as nonsteroidal mimics provided new VDR agonists and some antagonists with biological activity and possible therapeutical potential. Some of the D3 metabolites, including 20,23(OH)2D3 and 20(OH)D3 are able to inhibit RORE-mediated transactivation, as well as the interaction between the RORα/γ ligand-binding domain (LBD) with an LXXLL coactivator peptide. Our analysis of recently reported microarray data on vitamin D3 (D3) induced changes in cultured human keratinocytes indicated that D3 hydroxyderivatives stimulate the expression of genes involved in anti-aging activities. Furthermore, we noted upregulation of the kallikrein gene family by 1,25(OH)2D3 after 24-hour treatment, including stimulation of KLK6, KLK13, KLK3, KLK9, KLK5, KLK7, and KLK10. Also, after 6-hour incubation with 1,25(OH)2D3, the upregulation of KLK6, KLK13, and KLK3 was seen. Interestingly, ACEIs administered to hypertensive rats doubled the lifespan of these animals. In humans, ACEIs prevent hallmarks of aging, such as organ fibrosis and cardiac hypertrophy. We noted also that vitamin D3-hydroxyderivatives act against oxidative stress through upregulation of thioredoxin reductase (TXNRD1) and heme reductase-1 (HMOX-1) gene expression in keratinocytes treated for 24h. Another mechanism of anti-aging properties of inverse agonist RORa/γ is the resolution of inflammation caused by T helper (Th17) lymphocytes and switching the immune response into T regulatory (Treg) lymphocytes activation, with silencing of the inflammation state and reducing the inflammation process. The gene connected with inflammatory response, AKR1C3 (which encodes prostaglandin F synthase) is also strongly downregulated by 20,23(OH)2D3 in keratinocytes after incubation for 24 h. We suggest that vitamin D3 analogs, such as 20,23(OH)2D3, 1,25(OH)2D3, and 20(OH)D3 may have anti-aging properties through action on different pathways connected with DNA repair.

Octapeptide NOP-1 treatment delays yellowing in broccoli floret during low temperature storage

During postharvest life, floret yellowing confines broccoli's economic and nutritional worth. This study aimed to investigate the mechanisms activated by octapeptide NOP-1 (LKRYKRRL-NH2) treatment at 0, 250, 500, 750, or 1000 μM for retarding floret yellowing in broccoli during storage at 4 °C for 28 d. Our results revealed that the 750 μM NOP-1 treatment retarded chlorophyll degradation by suppressing pheophytinase (PPH) and pheophorbide a oxygenase (PaO) genes expression along with inhibiting Mg2+ dechelatase (MDC), and PPH enzymes activity. Besides, 750 μM NOP-1 treatment prevented jasmonic acid (JA) biosynthesis by suppressing phospholipase A (PLA), 13-lipoxygenase (13-LOX), allene oxide synthase (AOS), allene oxide cyclase (AOC), and OPDA reductase (OPR3) genes expression. Interestingly, 750 μM NOP-1 treatment maintained the membrane integrity in broccoli floret, evaluated by malondialdehyde (MDA) accumulation, which could be ascribed to the suppressing phosphatidylinositol 3-kinase (PI3K), phospholipase D (PLD), and 9-lipoxygenase (9-LOX) genes expression. Our results demonstrated that NOP-1, as an innovative, safe, and eco-friendly antisenescence octapeptide, significantly advances our technological proficiency to delay floret yellowing in broccoli during low temperature storage.

Effects on Steroid 5-Alpha Reductase Gene Expression of Thai Rice Bran Extracts and Molecular Dynamics Study on SRD5A2.

Simple SummaryDihydrotestosterone (DHT), the most potent androgen hormone, is an important aetiologic factor of androgenetic alopecia (AGA), or hair loss. Steroid 5-alpha reductases (SRD5As) increase DHT production in the scalp hair follicles, resulting in hair thinning and hair loss. Even though synthetic SRD5A inhibitors (finasteride and dutasteride) are effective in treating AGA, they cause adverse effects. This has led to an increased interest in alternative treatments from natural sources. The value of Thai rice bran has increased because several of its components may have use in AGA treatment. This study aimed to compare the suppression of the expression of SRD5A genes (type 1–3) exerted by several Thai rice bran extracts and investigate the interactional mechanism of their components towards SRD5A type 2. Tubtim Chumphae rice bran (TRB) had the highest sum of overall bioactive compounds. Among all extracts, the expression of SRD5A genes was suppressed by TRB as well as finasteride. In silico simulation showed that α-tocopherol had the greatest interaction with SRD5A type 2. Our findings identified α-tocopherol as the key bioactive in TRB; it could be developed as an anti-hair loss product.Steroid 5-alpha reductases (SRD5As) are responsible for the conversion of testosterone to dihydrotestosterone, a potent androgen, which is the aetiologic factor of androgenetic alopecia. This study aimed to compare the SRD5A gene expression suppression activity exerted by Thai rice bran extracts and their components and investigate the interactional mechanism between bioactive compounds and SRD5A2 using molecular dynamics (MD) simulation. Bran of Oryza sativa cv. Tubtim Chumphae (TRB), Yamuechaebia Morchor (YRB), Riceberry (RRB), and Malinil Surin (MRB), all rice milling by-products, was solvent-extracted. The ethanolic extract of TRB had the highest sum of overall bioactive compounds (γ-oryzanol; α-, β-, and γ-tocopherol; phenolics; and flavonoids). Among all extracts, TRB greatly downregulated the expression of SRD5A1, SRD5A2, and SRD5A3; there were no significant differences between TRB and finasteride regarding SRD5A suppression. The linear relationship and principal component analysis supported that the α-tocopherol content was correlated with the SRD5A suppression exerted by TRB. Furthermore, MD simulation demonstrated that α-tocopherol had the highest binding affinity towards SRD5A2 by interacting with residues Phe118 and Trp201. Our findings indicate that α-tocopherol effectively downregulates the expression of SRD5A genes and inhibits SRD5A2 activity, actions that are comparable to standard finasteride. TRB, a source of α-tocopherol, could be developed as an anti-hair loss product.

Inhibition of the mevalonate pathway improves myocardial fibrosis.

The mevalonate (MVA) pathway serves an important role in ventricular remodeling. Targeting the MVA pathway has protective effects against myocardial fibrosis. The present study aimed to investigate the mechanism behind these effects. Primary cultured cardiac fibroblasts from C57BL/6 mice were treated in vitro in 5 groups: i) negative control; ii) angiotensin II (Ang II) model (1x10-5 mol/l); iii) Ang II + rosuvastatin (ROS); iv) Ang II + alendronate (ALE); and v) Ang II + fasudil (FAS). Collagen and crystal violet staining were used to assess morphological changes in cardiac fibroblasts. Reverse transcription quantitative PCR and western blotting were used to analyze the expression of key signaling molecules involved in the MVA pathway. Collagen staining in the ALE, FAS, and ROS groups was weak compared with the Ang II group, while the rate of cell proliferation in the ROS, ALE, and FAS groups was slower compared with that in the Ang II group. In addition, the expression of key signaling molecules in the MVA pathway, including transforming growth factor-β1 (TGF-β1), heat shock protein 47 (HSP47), collagen type I α1 (COL1A1), vascular endothelial growth factor 2 (VEGF2) and fibroblast growth factor 2 (FGF2), was decreased in the FAS and ROS groups compared with the Ang II model. Compared with the Ang II group, 3-Hydroxy-3-Methylglutaryl-CoA reductase (HMGCR) gene expression was significantly lowered in the drug intervention groups, whereas farnesyl pyrophosphate synthase (FDPS) expression was downregulated in the ALE group, but elevated in the FAS and ROS groups. Compared with that in the Ang II group, ras homolog family member A (RhoA) expression was downregulated in the FAS and ROS groups, whilst mevalonate kinase expression was reduced in the ROS group. Protein expression of TGF-β1, COL1A1 and HSP47 were decreased following intervention with each of the three drugs compared with the Ang II group. Overall, rosuvastatin, aledronate and fasudil decreased the proliferation of myocardial fibroblasts and inhibited collagen synthesis. Rosuvastatin had the strongest protective effects against myocardial fibrosis compared with the other drugs tested, suggesting this to be a potential agent for the clinical treatment of cardiovascular disease.

Exogenous phytosulfokine α (PSKα) application delays senescence and relieves decay in strawberry fruit during cold storage by triggering extracellular ATP signaling and improving ROS scavenging system activity

During postharvest life, strawberry fruit as a perishable commodity exhibiting short shelf life suffers from fungal decay that severely limits its marketability and nutritional value. Herein, we investigated the mechanisms activated by exogenous phytosulfokine α (PSKα) application at 150 nM to delay senescence and relieve decay in strawberry fruit during storage at 4 °C for 18 days. Our results showed higher extracellular adenosine triphosphate (ATP) accumulation arising from lower apyrase 1 (APY1) gene expression in strawberry fruit treated with 150 nM PSKα during storage at 4 °C for 6 days may serve as a signaling molecule to promote NADPH oxidase activity to trigger signaling H2O2 accumulation giving rise to higher SUMO E3 ligase (SIZ1) gene expression during 18 days of storage at 4 °C. Also, higher superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) genes expression and enzymes activity associated with higher methionine sulfoxide reductase (MSR) and peroxiredoxin (Prx) genes expression in strawberry fruit treated with 150 nM PSKα during storage at 4 °C for 18 days may be ascribed to extracellular ATP signaling. As a result, promoting ROS scavenging systems activity by triggering extracellular ATP signaling in strawberry fruit treated with 150 nM PSKα may be responsible for higher maintenance of membrane integrity as shown by lower malondialdehyde (MDA) accumulation during 18 days of storage at 4 °C. Besides, strawberry fruits treated with 150 nM PSKα exhibited lower weight loss, total soluble solids, and chroma value concurrent with higher firmness, titrable acidity, L* value, and hue angle as quality attributes, which indicated delayed fruit senescence. Based on our findings, exogenous PSKα application unveils its potential to be used as a promising signaling biopeptide for senescence delay and decay alleviation in strawberry fruit during its postharvest life.

Molecular Cloning and Differential Gene Expression Analysis of 1-Deoxy-D-xylulose 5-Phosphate Synthase (DXS) in Andrographis paniculata (Burm. f) Nees.

Andrographis paniculata 1-deoxy-D-xylulose-5-phosphate synthase (ApDXS) gene (GenBank Accession No MG271749.1) was isolated and cloned from leaves for the first time. Expression of ApDXS gene was carried out in Escherichia coli Rosetta cells. Tissue-specific ApDXS gene expression by quantitative RT-PCR (qRT-PCR) revealed maximum fold expression in the leaves followed by stem and roots. Further, the differential gene expression profile of Jasmonic acid (JA)-elicited in vitro adventitious root cultures showed enhanced ApDXS expression compared to untreated control cultures. A. paniculata 3-hydroxy-3-methylglutaryl-coenzyme A reductase (ApHMGR) gene expression was also studied where it was up-regulated by JA elicitation but showed lower expression compared to ApDXS. The highest expression of both genes was found at 25 µm JA elicitation followed by 50 µm. HPLC data indicated that the transcription levels were correlated with increased andrographolide accumulation. The peak level of andrographolide accumulation was recorded at 25 μM JA (9.38-fold) followed by 50 µM JA (7.58-fold) in elicitation treatments. The in silico generated ApDXS 3D model revealed 98% expected amino acid residues in the favored and 2% in the allowed regions of the Ramachandran plot with 92% structural reliability. Further, prediction of conserved domains and essential amino acids [Arg (249, 252, 255), Asn (307) and Ser (247)] involved in ligand/inhibitor binding was carried out by in silico docking studies. Our present findings will generate genomic information and provide a blueprint for future studies of ApDXS and its role in diterpenoid biosynthesis in A. paniculata.Electronic supplementary materialThe online version of this article (10.1007/s12033-020-00287-3) contains supplementary material, which is available to authorized users.

Effects of Drip Irrigation Patterns and Biochar Addition on Soil Mineral Nitrogen and Microbial Regulation of Greenhouse

Drip irrigation and biochar amendment could affect the nitrogen form and transformation. Creating a deep understanding of the interacting effects of drip irrigation patterns and biochar on soil mineral nitrogen, as well as the key functional genes and microbial community involved in nitrogen transformation is helpful for improving facility agricultural management, increasing water and nitrogen use efficiency, and reducing the nitrate accumulation and groundwater pollution caused by nitrogen leaching. Four treatments [surface drip irrigation (D), insert drip irrigation (ID, insert depth 15 cm), surface drip irrigation +10 t·hm-2 of biochar (DB), and insert drip irrigation +10 t·hm-2 of biochar (IDB)] were conducted in a solar greenhouse, and non-rhizospheric and rhizospheric soils of pepper plants were studied. There was no effect of drip irrigation patterns and biochar on ammonium-nitrogen in the non-rhizospheric and rhizospheric soils. Compared with surface drip irrigation, insert drip irrigation decreased the nitrate-nitrogen concentration in the non-rhizosphere soil (P<0.05), but biochar addition weakened the difference. Biochar addition decreased the nitrate-nitrogen concentration in the rhizosphere soil under the same drip irrigation patterns. In the D treatment, biochar significantly decreased the number of copies of AOA, AOB, and nirK genes in the non-rhizospheric soil, and AOA gene copies in the rhizospheric soil (P<0.05); however, there was an increase in the number of copies of AOB and nirK genes in the rhizospheric soil of the D and ID treatments (P<0.05). Based on the structural equation model (SEM), in the non-rhizospheric and rhizospheric soils, pH and electrical conductivity were the environmental factors with the greatest influence on the ammonium-nitrogen and nitrate concentrations, respectively, and the gene copy number of AOB was the biotic factor with the greatest influence on the nitrate-nitrogen concentration. Based on PICRUSt, the γ-Proteobacteria contributed mostly to ammonia monooxygenase gene (K10945) expression, whereas the α-Proteobacteria, especially the rhizobia members, contributed mostly to nitrite reductase gene (K00368) expression. Biochar addition regulated the bacterial community structure that participated in K10945 gene expression in the non-rhizospheric soil and K00368 gene expression in the rhizospheric soil (P<0.05). Overall, biochar addition contributed more to nitrate-nitrogen and microbial mineral nitrogen-transformation processes in the agricultural soil than did the drip irrigation patterns.