Evaluation of the intracellular lipid-lowering effect of polyphenols extract from highland barley in HepG2 cells

Active ingredients from highland barley have received considerable attention as natural products for developing treatments and dietary supplements against obesity.In practical application,the research of food combinations is more significant than a specific food component.This study investigated the lipid-lowering effect of highland barley polyphenols via lipase assay in vitro and HepG2 cells induced by oleic acid(OA).Five indexes,triglyceride(TG),total cholesterol(T-CHO),low density lipoprotein-cholesterol(LDL-C),aspartate aminotransferase(AST),and alanine aminotransferase(ALT),were used to evaluate the lipidlowering effect of highland barley extract.We also preliminary studied the lipid-lowering mechanism by Realtime fluorescent quantitative polymerase chain reaction(q PCR).The results indicated that highland barley extract contains many components with lipid-lowering effects,such as hyperoside and scoparone.In vitro,the lipase assay showed an 18.4%lipase inhibition rate when the additive contents of highland barley extract were 100μg/m L.The intracellular lipid-lowering effect of highland barley extract was examined using 0.25 mmol/L OA-induced HepG2 cells.The results showed that intracellular TG,LDL-C,and T-CHO content decreased by 34.4%,51.2%,and 18.4%,respectively.ALT and AST decreased by 51.6%and 20.7%compared with the untreated hyperlipidemic HepG2 cells.q PCR results showed that highland barley polyphenols could up-regulation the expression of lipid metabolism-related genes such as PPARγand Fabp4.

Antibacterial mechanism of kojic acid and tea polyphenols against Escherichia coli O157:H7 through transcriptomic analysis

Escherichia coli O157:H7 is one of the major foodborne pathogenic bacterial that cause infectious diseases in humans.The previous found that a combination of kojic acid and tea polyphenols exhibited better activity against E.coli O157:H7 than using either alone.This study aimed to explore responses underlying the antibacterial mechanisms of kojic acid and tea polyphenols from the gene level.The functional enrichment analysis by comparing kojic acid and tea polyphenols individually or synergistically against E.coli O157:H7 found that acid resistance systems in kojic acid were activated,and the cell membrane and genomic DNA were destructed in the cells,resulting in“oxygen starvation”.The oxidative stress response triggered by tea polyphenols inhibited both sulfur uptake and the synthesis of ATP,which affected the bacteria's life metabolic process.Interestingly,we found that kojic acid combined with tea polyphenols hindered the uptake of iron that played an essential role in the synthesis of DNA,respiration,tricarboxylic acid cycle.The results suggested that the iron uptake pathways may represent a novel approach for kojic acid and tea polyphenols synergistically against E.coli O157:H7 and provided a theoretical basis for bacterial pathogen control in the food industry.

Utilization of pH-driven methods to fortify nanoemulsions with multiple polyphenols

Simple but effective methods are required to incorporate multiple bioactive polyphenols into delivery systems to increase their dispersibility,stability and bioavailability.We developed and tested three p Hdriven protocols for creating nanoemulsions loaded with multiple lipophilic polyphenols.These protocols differed in how the different polyphenols were incorporated into the nanoemulsions.The impact of these three methods on the formation,properties,and gastrointestinal fate of nanoemulsions loaded with curcumin,resveratrol,and quercetin was investigated.The three methods produced nanoemulsions with similar initial particle properties:droplet diameters(0.15,0.16,and 0.15μm)and zeta-potentials(–59,–58,and–58 m V),respectively.However,the average encapsulation efficiencies(82%,88%,and 61%),gastrointestinal stabilities(83%,97%,and 29%)and bioaccessibilities(77%,90%,and 73%)for curcumin,resveratrol,and quercetin were somewhat different.In particular,more quercetin degradation occurred using the approach that held it under alkaline conditions for extended periods.In general,the p H-driven method provides researchers with a versatile approach of incorporating multiple polyphenols with different characteristics into functional food and beverages using a simple and inexpensive method.

Protective effects of oleic acid and polyphenols in extra virgin olive oil on cardiovascular diseases

The Mediterranean diet has long been recognized as one of the most effective ways to prevent and improve cardiovascular disease.Extra virgin olive oil(EVOO)is the typical sources of fat in the Mediterranean diet which have been shown to have noteworthy nutritional value and positive impact on human health.It is worth noting that EVOO owes its superior nutritional value to its bioactive composition.The main component of EVOO is monounsaturated fatty acids(MUFAs)in the form of oleic acid.Oleic acid accounts for up to 70%-80%of EVOO.Secondly,EVOO contains approximately more than 30 phenolic compounds,of which HT is essential for the protection against cardiovascular diseases.In this review,we focused on the potential mechanisms of oleic acid and polyphenols combat cardiovascular diseases risk in terms of oxidative stress,inflammation,blood pressure,endothelial function and cholesterol.This review might provide a reference for the studies on cardiovascular protective effects of EVOO.

The modulatory effect of oolong tea polyphenols on intestinal flora and hypothalamus gene expression in a circadian rhythm disturbance mouse model

The interaction between host circadian rhythm and gut microbes through the gut-brain axis provides new clues for tea polyphenols to improve host health.Our present research showed that oolong tea polyphenols(OTP)improved the structural disorder of the intestinal flora caused by continuous darkness,thereby modulating the production of metabolites related to pyruvate metabolism,glycolysis/gluconeogenesis,and tryptophan metabolism to alleviate the steady-state imbalance.After fecal microbiota transplantation from the OTP group,the single-cell transcriptomic analysis revealed that OTP significantly increased the number of hypothalamus cell clusters,up-regulated the number of astrocytes and fibroblasts,and enhanced the expression of circadian rhythm genes Cry2,Per3,Bhlhe41,Nr1d1,Nr1d2,Dbp and Rorb in hypothalamic cells.Our results confirmed that OTP can actively improve the intestinal environmental state as well as internal/peripheral circadian rhythm disorders and cognitive impairment,with potential prebiotic functional characteristics to notably contribute to host health.

Sugarcane leaves-derived polyphenols alleviate metabolic syndrome and modulate gut microbiota of ob/ob mice

Sugarcane leaves-derived polyphenols(SLP)have been demonstrated to have diverse health-promoting benefits,but the mechanism of action has not been fully elucidated.This study aimed to investigate the anti-metabolic disease effects of SLP and the underlying mechanisms in mice.In the current study,we prepared the SLP mainly consisting of three flavonoid glycosides,three phenol derivatives,and two lignans including one new compound,and further demonstrated that SLP reduced body weight gain and fat accumulation,improved glucose and lipid metabolism disorders,ameliorated hepatic steatosis,and regulated short-chain fatty acids(SCFAs)production and secondary bile acids metabolism in ob/ob mice.Notably,SLP largely altered the gut microbiota composition,especially enriching the commensal bacteria Akkermansia muciniphila and Bacteroides acidifaciens.Oral gavage with the above two strains ameliorated metabolic syndrome(MetS),regulated secondary bile acid metabolism,and increased the production of SCFAs in high-fat diet(HFD)-induced obese mice.These results demonstrated that SLP could be used as a prebiotic to attenuate MetS via regulating gut microbiota composition and further activating the secondary bile acids-mediated gut-adipose axis.

Emerging and multifaceted potential contributions of polyphenols in the management of type 2 diabetes mellitus

Type 2 diabetes mellitus(T2DM)is recognized as a serious public health concern with a considerable impact on human life,long-term health expenditures,and substantial health losses.In this context,the use of dietary polyphenols to prevent and manage T2DM is widely documented.These dietary compounds exert their beneficial effects through several actions,including the protection of pancreatic islet β-cell,the antioxidant capacities of these molecules,their effects on insulin secretion and actions,the regulation of intestinal microbiota,and their contribution to ameliorate diabetic complications,particularly those of vascular origin.In the present review,we intend to highlight these multifaceted actions and the molecular mechanisms by which these plant-derived secondary metabolites exert their beneficial effects on type 2 diabetes patients.

Changes in Polyphenols and Antioxidant Activities of Yingshan Yunwu Tea during Digestion in Vitro

[Objectives]To explore the change rule of polyphenol content and antioxidant activity of coarse old leaves of Yingshan Yunwu Tea in the process of human digestion.[Methods]The coarse and old leaves of Yunwu tea in Yingshan,Huanggang,Hubei Province were selected as the research object,and their digestion in vitro was simulated.The total polyphenol content was determined by Folin-phenol reagent colorimetric method,and the DPPH radical scavenging activity and total antioxidant activity were determined.[Results]After simulated gastrointestinal digestion in vitro,the polyphenol content and antioxidant activity of coarse old leaf tea soup showed a downward trend.After gastrointestinal digestion,the polyphenol content in tea infusion separately decreased by 31.8%and 8.5%;the scavenging rate of DPPH free radical was 97%before digestion,decreased to 92%after gastric digestion and 65%after intestinal digestion,which decreased by 5%and 27%,respectively;after gastrointestinal digestion,the total antioxidant capacity of tea soup decreased by 4.7%and 3.1%,respectively.[Conclusions]This study provided a reference for the development and application of coarse old leaves of Yingshan Yunwu tea,and provided a reference for the nutritional value evaluation and comprehensive utilization of coarse old leaves,so as to make the best use of coarse tea leaves and reduce the waste of resources.

Exploring the mechanism of hypolipidemic effect of polyphenols in wolfberry based on network pharmacology and molecular docking

To explore the mechanism of hypolipidemic action of wolfberry polyphenols by using network pharmacology and molecular docking.The active ingredients and targets of wolfberry were searched by TCMSP,and the Cytoscape 3.9.0 software was used to construct a“wolfberry component-target”network.The Gene Cards database was used to screen the hyperlipidemic targets and intersect them with the active targets of wolfberry to construct the PPI network using the STRING platform.The gene ontology(GO)function and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis of the core targets were carried out on the Metascape platform,and molecular docking of the active ingredients to the core targets was performed using AutoDockTools software.A total of 33 active ingredients and 173 potential targets of wolfberry were screened,including 99 targets related to hyperlipidemia.The results of the analysis of 99 intersecting targets with the components of wolfberry identified the core active ingredients as quercetin,glycitein and atropine.The binding of the major components of wolfberry,including the polyphenolic compounds quercetin and glycitein,as well as atropine to the key targets AKT1,IL6 and TNF may be important mechanisms for the hypolipidemic therapeutic effect.GO functional enrichment analysis involves biological processes,cellular components,and molecular functions.The KEGG pathway enrichment analysis mainly involves the AGE-RAGE signaling pathway,fluid shear stress,and TNF signaling pathway.Molecular docking validated the good binding activity of the targets to the active ingredients.The binding of atropine and the polyphenolic compounds quercetin and glycitein to the key targets AKT1,IL6 and TNF may be an important mechanism for the hypolipidemic therapeutic effect of wolfberry.

Lonicera caerulea polyphenols inhibit fat absorption by regulating Nrf2-ARE pathway mediated epithelial barrier dysfunction and special microbiota

Scope:High-fat diet(HFD)induces imbalance in the small intestine environment,where fat digestion and absorption mainly take place.This study aimed to elucidate the mechanisms by which Lonicera caerulea polyphenols(LCP)might inhibit fat absorption,from the perspective of small intestine microbiota and epithelial barrier integrity.Methods and results:Male Sprague-Dawley rats were given HFD with or without co-administration of LCP for 8 weeks.The results showed that LCP supplementation significantly decreased the levels of serum triglycerides(TG),total cholesterol(TC),and low-density lipoprotein cholesterol(LDL-C),and increased the contents of fecal sterols,in HFD rats.LCP also inhibited the dysfunction of the small intestine epithelial barrier,via alleviating the oxidative stress activated by Nrf2-ARE pathway,and by modulating the expressions of pro-inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),cyclooxygenase-2(COX-2),nuclear factor kappa-B p65(NF-κB p65)and inducible nitric oxide synthase(iNOS)in the small intestine.Additionally,LCP administration restored the balance in small intestine microbiota and increased the abundance of the specific bacteria,such as Lactobacillus,involved in fat absorption.Conclusion:Our results demonstrated that LCP may be beneficial to inhibit fat absorption.The mechanism seems to be associated with the protection of the epithelial barrier integrity and the modulation of specific bacteria in the small intestine.

Antioxidant Activities and Characterization of Polyphenols from Selected Northern Thai Rice Husks: Relation with Seed Attributes

Rice production generates a significant amount of agricultural waste. This study aimed to give results related to the existence of antioxidant phenols in agricultural waste of selected Northern Thai rice varieties. The antioxidant activities, contents of total flavonoids and phenolic compounds in the ethanolic rice husk extract were evaluated. The highest antioxidant activities were found in the variety PES1CMU, with 2,2’-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid and 2,2-diphenyl-1-picrylhydrazyl as 679.66 and 4.16 mmol/(L·g) trolox equivalent, respectively, ferric reducing antioxidant power as 0.87 mmol/(L·g) Fe2+, total phenolic content as 29.90 mmol/(L·g) gallic acid and total flavonoid content as 12.16 mg/g catechin equivalent. Polyphenol compounds were identified mainly by standard polyphenols using the liquid chromatography mass spectrometry, with the highest contents of phytic acid, o-coumaric acid, naringin and kaempferol. The non-glutenous and wetland ecotypes of rice husk samples were the richest in antioxidant activities and polyphenol contents characterized by using principal component analysis. The glutenous rice husk contained higher antioxidant activities than the rest. Interestingly, quercetin is a significant phenolic compound that positively correlated with the overall antioxidant activities of rice husk. This finding will be relevant for future application of rice husk antioxidant components in the production of functional ingredients as well as for the food and pharmaceutical industries.

Tea Polyphenols Reduce Inflammation of Orbital Fibroblasts in Graves’Ophthalmopathy via the NF-κB/NLRP3 Pathway

Objective:This study aimed to explore the effects of tea polyphenols(TP)on inflammation of orbital fibroblasts in Graves’ophthalmopathy(GO)and to provide new ideas for GO treatment.Methods:Primary orbital fibroblasts were extracted from orbital adipose/connective tissues of patients with and without GO.Real-time quantitative PCR(RT-qPCR)was used to detect the expression of interleukin(IL)-6,IL-1β,and monocyte chemotactic protein(MCP)-1 in non-GO and GO orbital fibroblasts.The CCK-8 assay was used to determine the appropriate concentration of TP for subsequent experiments.RT-qPCR and enzyme-linked immunosorbent assay(ELISA)were performed to investigate the effects of TP on lipopolysaccharide(LPS)-induced production of inflammatory cytokines.Nuclear factor-κB(NF-κB)expression was measured using Western blotting analysis.NOD-like receptor 3(NLRP3)expression was detected using both Western blotting analysis and immunofluorescence staining.Results:The mRNA levels of IL-6,IL-1β,and MCP-1 in GO orbital fibroblasts were significantly higher than those in non-GO cells.TP treatment significantly inhibited LPS-induced production of inflammatory factors,including IL-6,IL-1β,and MCP-1.TP also inhibited the expression levels of NF-κB and NLRP3.Inflammation in the GO orbital fibroblasts was higher than that in non-GO cells.TP inhibited the production of inflammatory cytokines in GO orbital fibroblasts in vitro through the NF-κB/NLRP3 pathway.Conclusion:These findings suggest that TP may have a potential role in GO treatment.

Berry综合征患儿的围手术期护理

总结1例Berry综合征患儿的围手术期护理体会。术前针对患儿年龄小、病情危重,采取积极营养支持以尽早达到手术所需体重,维持循环稳定以确保动脉导管持续开放,严格容量管理以降低肺动脉高压等护理措施;术后针对患儿手术复杂、病情危重,采取延迟关胸护理、呼吸功能支持、并发症管理、出院健康指导等护理措施。经过精心护理后,患儿顺利出院。出院后随访1个月余,患儿心功能恢复良好。

Variation of Polyphenols, Anthocyanins and Antioxidant Power in the Strawberry Grape (<i>Vitis labrusca</i>) after Simulated Gastro-Intestinal Transit and Evaluation of <i>in Vitro</i>Antimicrobial Activity

The influence of a simulated digestive process on some biochemical and biological aspects of strawberry grape (Vitis labrusca) was investigated. The amount of total polyphenols and anthocyanins as well as the antioxidant power were evaluated. Results evidenced that the simulated gastrointestinal transit caused a decrease of the polyphenols content and total anthocyanins;these last, however, were more resistant than polyphenols, decreasing only of 50% respect to the initial value (31.50 μg/ml of extract). The extract exhibited an excellent antioxidant power (EC50 3.8 mg/ml), which decreased of about four times after the simulated gastrointestinal transit. The antimicrobial activity of the extract, evaluated against three Escherichia coli, Pseudomonas aeruginosa and Bacillus cereus pathogen strains was enhanced by the simulated digestion, with an increase of the inhibition halo.

Polyphenols of the Hyperhalophyte Halocnemum strobilaceum (Pall.) M. Bieb. and Antioxidant Activity

The use of plant resources in obtaining medicines and locally useful species is an important step in the creation of a sustainable raw material base. The promising use of species of the Chenopodiaceae family, as a medicinal and food raw material, makes it relevant to investigate new promising sources of polyphenols to identify the chemical structure of compounds and their biological activity. Because the extract from Halocnemum strobilaceum has high antiradical activity, it seems important to study the possibility of correction of disturbances in oncological, diabetic and other non-infectious patients and consider them as promising drugs. In the first identified 5 polyphenols from Halocnemum strobilaceum, the obtained analyses show that this desert plant will allow creating in the future of a new range of promising biologically active substances, as well as using effectively the plant cover around the Aral.

Effect of Sodium Carbonate on Extraction by Aqueous Decoction of Total Polyphenols from Crushed and Whole Leaves of Combretum micranthum

Sodium bicarbonate is sometimes used to aid in the extraction of total polyphenols. Its main effect is to increase the pH of the extraction solution. Raising the pH can cause changes in the chemical structure of polyphenols. This can lead to variations in their biological properties, solubility and stability. This work studied the effect of sodium carbonate on the extraction by aqueous decoction of total polyphenols from the leaves of Combretum micranthum. The content of total phenolic compounds in the extracts was estimated by the Folin-Ciocalteu method. The color of the samples was measured using a colorimeter (type: KONICA MINOLTA. Japan) based on the CIELAB color system. The results obtained were subjected to a one-way ANOVA analysis of variance with R software version 3.2.4 Revised (2018) and Minitab-18 software. The results reveal a drop in the concentration of extracted polyphenols proportional to the addition of sodium carbonate, i.e. a drop from 3.30 to 1.04 mg·AG·100 g-1 of extract on whole leaves and 3.921 to 2.551 mg·AG·100 g-1 extract on crushed leaves. On the other hand, the intensity of the coloring of the extracts increases significantly with the addition of sodium carbonate from 0.0 g·L -1 to 0.666 g·L-1.

Polyphenols Extracted from Enteromorpha clathrata Induce Apoptosis in Hepa1-6 Cell by Activating the Mitochondrial Apoptosis Signaling Pathways

Green alga Enteromorpha clathrata(E.clathrata)contains a variety of bioactive compounds,including polysaccharides,polyphenols and fat-soluble pigments etc.,among which polyphenols exhibit a wide range of medicinal properties.E.clathrata polyphenols(ECPs)have shown various biological activities such as antioxidant,anti-inflammatory and antidiabetic effects;however,the potential of ECPs as an anti-cancer reagent remains unclear.The aim of this study was to investigate the anti-tumor activity and underlying mechanisms of ECPs on hepatocellular carcinoma.The cytotoxicity of Hepa1-6 cells was determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)and lactate dehydrogenase(LDH)assay.Flow cytometry and fluorescence microscope analysis of cell apoptosis after annexin V-fluorescein isothiocyanate(FITC)/propidium iodide(PI)staining.2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)assay was used for intracellular reactive oxygen species(ROS)detection.caspase-9 activity was determined using cspase-9 colorimetric assay.Mitochondrial transmembrane potential(Δψm)was measured using JC-1.Western blot and quantitative real-time PCR(qPCR)were used to assess the expressions of the apoptosis regulators Bax,Bcl-2,cytochrome c and caspase-3.It was found that ECPs showed a dose-dependent cytotoxicity against Hepa1-6 cells by inducing apoptosis.The apoptosis in ECPs-treated Hepa1-6 cells was accompanied by the loss of mitochondrial membrane potential,elevated ROS generation,increased release of mitochondrial cytochrome c,and up-regulation of caspase-9 and caspase-3.The expressions of Bax(pro-apoptotic molecule)and Bcl-2(apoptosis suppressor)were up-regulated and down-regulated,respectively,at both mRNA and protein levels.These molecular alterations revealed that ECPs caused apoptosis of cells through the mitochondrial pathway,suggesting that ECPs are potential candidates to be developed for liver cancer treatment.

Research Progress on the Therapeutic Mechanism of Tea Polyphenols in Neurodegenerative Diseases

In recent years, the incidence of neurodegenerative diseases, mainly Alzheimer’s disease, Parkinson’s disease, vascular dementia, and cerebral ischemia, has been rising gradually, which has a serious impact on the physiological state and quality of life of human beings in old age, and the current clinical drugs are unsatisfactory in terms of therapeutic efficacy and healing, which has made this kind of diseases become a social medical problem. Tea polyphenols are the main functional components of tea and have great potential in neuroprotection. In this paper, we review the research on tea polyphenols in neurodegenerative diseases, with the aim of providing a new entry point for the treatment of neurodegenerative diseases.

Plant-inspired biomimetic hybrid PVDF membrane co-deposited by tea polyphenols and 3-amino-propyl-triethoxysilane for high-efficiency oilin-water emulsion separation

Membrane pollution caused by separating oily wastewater is a big challenge for membrane separation technology.Recently,plant-/mussel-inspired interface chemistry has received more and more attention.Herein,a high antifouling poly(vinylidene fluoride)(PVDF)membrane,coated with tea polyphenols(TP,extracted from green tea)and 3-amino-propyl-triethoxysilane(APTES),was developed to purify oil-inwater emulsions.ATR-FTIR,XPS and SEM were used to demonstrate the evolution of surface biomimetic hybrid coatings.The performances of the developed membranes were investigated by pure water permeability and oil rejection for various surfactant-stabilized oil-in-water emulsions.The experimental results revealed that the membrane deposited with a mass ratio of 0.1/0.2 exhibited ultrahigh pure water permeability(14570 L·m^(-2)·h^(-1)·bar^(-1),1 bar=0.1 MPa)and isooctane-in-water emulsion permeability(5391 L·m^(-2)·h^(-1)·bar^(-1))with high separation efficiency(>98.9%).Even treated in harsh environment(acidic,alkaline and saline)for seven days,the membrane still maintained considerable underwater oleophobic property(148°–153°).The fabricated plant-inspired biomimetic hybrid membranes with excellent performances light a broad application prospect in the field of oily wastewater treatment.

Extraction of Flavonoids and Polyphenols from Coronatinetreated Mulberry Leaves and Evaluation of Their Antibacterial Activity against Pseudomonas syringae

Flavonoids and polyphenols are the main secondary metabelites in mulberry leaves, which have multiple biological functions such as antibacterial activity, free radical scavenging activity and antioxidant activity. In this paper, the parameters for the extraction of flavonoids and palyphenols from eoronatine （ COR）- treated mulberry leaves were optimized, and the components of the extracts were analyzed by high performance liquid chromatography （ HPLC）, and their antibacterial activity against Pseudomonas syringae, which causes mulberry bacterial blight, was evaluated in vitro to explore the mechanism how COR acted as a plant immune elicitor to improve the resistance of mulberry to bacterial blight. The results showed that ethanol concentration of 70%, extraction temperature of 80 ℃, extraction duration of 6 h and solid-liquid ratio of 1 ： 40 were the optimal parameters for the extraction of flavonoids and polyphenols from mulberry leaves. Flavonoids and polyphenols were mostly distributed in ethyl acetate and n-butyl alcohol fractions which showed better antibacterial activity than petroleum ether, dichloromethane and water fractions. Meanwhile, the antibacterial activity of ethyl acetate and n-butyl alcohol fractions of COR-treated mulberry leaves were more evident than that of water-treated mulberry leaves. Scanning electron microscopy showed that flavonoids and pelyphenols inhibited the growth of pathogenic bacteria by changing cell morphology, damaging cell wall structure and unbalancing cell osmotic pressure. HPLC analysis confirmed the contents of morin, quercetin, quereitrin, kaempferol, chlorogenic acid and rutin were increased to a different extent in COR-treated mulberry leaves. So it can be concluded that the increased contents of flavonoids and polyphenols induced by COR may be one of the reasons for the enhanced resistance to mulberry bacterial blight.