Health effects of fruit juices and beverages with varying degrees of processing

The degree of processing is rarely considered an independent factor in the health effects of fruit juices and beverages(FJBs)consumption.In fact,the consumption of ultra-processed foods has been shown to pose health risks.In this study,we first integrated 4 systems used to classify the degree of food processing and then classified FJBs into three major categories,low(minimal),moderate and high.Second,we compared the differences in attitudes towards FJBs in dietary guidelines.Third,we integrated the results of existing epidemiological surveys,randomized controlled trials,and animal experiments to explore the health risks associated with consuming FJBs.Deepening the processing of FJBs has been found to lead to an increased risk of diseases.Dietary pattern,nutrients,addition agents and consumer preferences may be influential factors.Finally,we investigated whether there were any changes in the health benefits of 100%fruit juices produced by different processing methods.In conclusion,minimally/moderately processed 100%fruit juices provide more health benefits than highly processed fruit beverages.The results support the need to consider the extent of FJBs processing in future studies to adjust official nutritional recommendations for beverage consumption.

Efficient large fragment deletion in plants:double pairs of sgRNAs are better than dual sgRNAs

Dear Editor,Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)is a powerful and versatile gene editing system that has been extensively utilized in various animals and plants,which holds enormous potential and value for scientific research and breeding.However,single-targeted CRISPR can only induce a few base deletions,insertions,or substitution.Ideally,thesemutations result in premature termination of the protein encoded by the target gene,leading to a loss of function[1].

Development ofκ-carrageenan hydrogels with mechanically stronger structures via a solvent-replacement method

Strongκ-carrageenan(KC)hydrogels were fabricated via solvent replacement with sorbitol,and the effects of KC mass fraction and solvent replacement on the structural characteristics encapsulation capability of the hydrogels were evaluated.Microstructural observation showed that the 3D network structures of hydrogels exhibited a complete and continuous skeleton.FTIR spectra of KC hydrogels revealed the formation of intermolecular hydrogen bonds after sorbitol replacement.The stability against heating and freeze-thawing of hydrogels was enhanced due to the addition of sorbitol and the rise in KC mass fraction.The hydrogel with 1.5 wt%KC after sorbitol replacement presented the best stability.Frequency sweep tests suggested that storage modulus of the samples were influenced by sorbitol replacement and KC concentration.Swelling tests revealed that the hydrogels after replacement with a higher KC content(1.25,1.50 wt%)presented higher swelling capacity,and they were more stable in alkaline and acidic solutions.When epigallocatechin gallate(EGCG)was incorporated within the hydrogels,the hydrogels after sorbitol replacement offered higher protection capability.The information obtained in this study indicated that sorbitol replacement strengthened KC hydrogels,and they could act more appropriately as accountable carriers for bioactives.

Research progress of protein haze in white wines

Protein haze was one of the main causes of the instability of white wines. Proteins that caused haze or precipitation in white wines mainly came from grape fruits, and their compositions and contents were affected by many factors such as fruit diseases, harvesting methods and water stress. Unstable wine proteins were usually pathogenesis-related(PR)proteins of grapes, mainly chitinases and thaumatin-like proteins(TLPs), which had lower isoelectric point(pI)and smaller molecular weight, and were highly resistant to the low pH values of wines and the protease hydrolysis during fermentation. At present, the technology of protein stabilization and clarification in white wines mainly included bentonite fining, heat treatment, enzymatic hydrolysis, polysaccharide treatment and ultrafiltration methods. Among them, the most commonly used method was bentonite treatment. In this paper, the research progresses of the origin, mechanism and influencing factors of the unstable proteins in white wines were summarized, and the applications, advantages and disadvantages of various clarification techniques were also concluded, in order to provide some support for the theoretical and technological research of the protein stability in white wines in the future.

Effects of heat treatment on the structure, digestive property, and absorptivity of holoferritin

Ferritin,as an iron storage protein,has been considered to be a well-utilized iron supplement.However,the typical thermal processing of food rich in ferritin may affect the structure and function of ferritin as an iron supplement.Here,a plant ferritin(soybean seed ferritin,SSF)and an animal ferritin(donkey spleen ferritin,DSF)were used to analyze the changes in fundamental structure and iron content after thermal treatments(68°C for 30 min,100°C for 10 min).Then,SSF and DSF after thermal treatment were administered intragastrically to mice to further evaluate its digestive stability and absorptivity after thermal processing.Results showed the secondary structure,oligomeric states,iron content,and digestive stability of DSF were maintained better than that of SSF after thermal treatments,indicating that DSF has a higher thermostability than SSF.Both SSF and DSF after thermal treatment exhibited higher absorptivity than untreated ferritins.SSF showed higher absorptivity than DSF whether heated or not.

The synergistic effect of dietary cholesterol with fruit tannins in forming kidney stones

Prevalence of kidney stones has increased continously over several decades worldwide,the major causes of which are largely unknown.To explore the dietary causes of kidney stones,and reveal mechanisms underlying dietary risk factors inducing kidney stones,animal experiments using mice as the disease model were performed.Eight-week old male CD-1 mice were treated by ethylene glycol,cholesterol or/and apple tannins for 3 d,respectively.In the present study,the crystalline analysis in urine and kidney tissues,HE staining kidney sections as well as observation of micro-stones,tannins and cholesterol deposition in kidneys of mice in different groups were conducted.We found that gavage with ethylene glycol,cholesterol and tannins resulted in mice urine solute supersaturation in renal tubules and forming kidney stones.Significant cholesterol and tannin deposits in mouse kidney were observed by laser confocal microscopy and crystals were shown either adhered with or codeposited with cholesterol and tannin deposits.The primary crystals were found in renal cortex,medullar,especially papilla in the kidney sections under polarized microscope.These findings demonstrate that interaction of cholesterol and tannins in kidney plays a critical role in the formation of kidney stones.

Purification and Characterization of Three Alkaline Endopolygalacturonases from a Newly Isolated Bacillus gibsonii

A newly isolated Bacillus gibsonii,designated as S-2(CGMCC1215),was cultivated for production of alkaline pectinases utilizing sugar beet pulp as growth substrate.Purification of three alkaline endopolygalacturonases(endoPGs)from the crude pectinases extract was carried out by ultra-filtration,ammonium sulphate fractionation and ion-exchange chromatography,and their enzyme activities characterized.The three purified alkaline endoPGs,designated as S-I,S-II,and S-III,had a molecular weight about38kDa as determined by SDS-PAGE.The Km value and optimal temperature for optimal enzyme activities of S-I,S-II and S-III were1.2mg/mL and60℃,0.9mg/mL and55℃,1.1mg/mL and60℃,respectively.Their best performances were given at an optimal pH10.5,and sodium polygalacturonate was found to be the best substrate.The isoelectric points of S-I,S-II and S-III were5.4,7.4,and8.2,respectively.Surfactants of Tween-80and Tween-20and metal ions such as Mg2+and Ca2+stimulated the activity of S-I,S-II and S-III,whereas S-III was inhibited by Ca2+,and Mn2+and Zn2+ions inhibited the activity of the three enzymes.

A NAC transcription factor, NOR-like1, is a new positive regulator of tomato fruit ripening

Ripening of the model fruit tomato(Solanum lycopersicum)is controlled by a transcription factor network including NAC(NAM,ATAF1/2,and CUC2)domain proteins such as No-ripening(NOR),SlNAC1,and SlNAC4,but very little is known about the NAC targets or how they regulate ripening.Here,we conducted a systematic search of fruit-expressed NAC genes and showed that silencing NOR-like1(Solyc07g063420)using virus-induced gene silencing(VIGS)inhibited specific aspects of ripening.Ripening initiation was delayed by 14 days when NOR-like1 function was inactivated by CRISPR/Cas9 and fruits showed obviously reduced ethylene production,retarded softening and chlorophyll loss,and reduced lycopene accumulation.RNA-sequencing profiling and gene promoter analysis suggested that genes involved in ethylene biosynthesis(SlACS2,SlACS4),color formation(SlGgpps2,SlSGR1),and cell wall metabolism(SlPG2a,SlPL,SlCEL2,and SlEXP1)are direct targets of NOR-like1.Electrophoretic mobility shift assays(EMSA),chromatin immunoprecipitation-quantitative PCR(ChIP-qPCR),and dual-luciferase reporter assay(DLR)confirmed that NOR-like1 bound to the promoters of these genes both in vitro and in vivo,and activated their expression.Our findings demonstrate that NOR-like1 is a new positive regulator of tomato fruit ripening,with an important role in the transcriptional regulatory network.

Evaluation of growth characteristics of Aspergillus parasiticus inoculated in different culture media by shortwave infrared(SWIR) hyperspectral imaging

The growth characteristics of Aspergillus parasitic us incubated on two culture media were ex-amined using shortwave infrared(SWIR,1000-2500 nm)hyperspectral imaging(HSI)in this work.HSI images of the A.parasiticus colonies growing on rose bengal medium(RBM)and maize agar medium(MAM)were recorded daily for 6 days.The growth phases of A.parasiticus were indicated through the pixel number and average spectra of colonies.On score plot of the first principal component(PC1)and PC2,four growth zones with varying mycelium densities were identified.Eight characteristic wavelengths(1095,1145,1195,1279,1442,1655,1834 and 1929 nm)were selected from PC1 loading,average spectra of each colony as well as each growth zone.F urthermore,support vector machine(S VM)classifier based on the eight wavelengths was built,and the classification accuracies for the four zones(from outer to inner zones)on the colonies on RBM were 99.77%,9935%,99.75%and 99.60%and 99.77%,9939%,99.31%and 98.22%for colonies on MAM.In addition,a new score plot of PC2 and PC3 was used to differ-entiate the colonies incubated on RBM and MAM for 6 days.Then characteristic wavelengths of 1067,1195,1279,1369,1459,1694,1834 and 1929 nm were selected from the loading of PC2 and PCg.Based on them,a new SVM model was developed to diferentiate colonies on RBM and MAM with accuracy of 100.00%and 9999%,respectively.In conclusion,SWIR hyperspectral image is a powerful tool for evaluation of growth characteristics of A.parasiticus incubated in diferent culture media.

Mutagenesis of SlNAC4 by CRISPR/Cas9 alters gene expression and softening of ripening tomato fruit

Softening is one of the key fruit quality traits,which results from the selective expression of cell wall metabolism genes during ripening.The identification of transcription factors(TFs)that regulate fruit softening is an important field in order to understand and control fruit softening.In tomato,NAC(NAM,ATAF,and CUC)TFs members have been demonstrated to be involved in fruit ripening regulation,including NAC-NOR(nonripening),NOR-like1,SlNAC4,SlNAC1.Here,we generated slnac4 mutant knockout(CR-SlNAC4)tomato plant by a clustered regularly interspaced short palindromic repeats genomic targeting system(CRISPR/Cas9)and SlNAC4 overexpressing(OE-SlNAC4)plant.In addition to confirming the previously reported results that SlNAC4 positively regulates fruit ripening,we found that SlNAC4 has a strong effect on tomato fruit softening.Compared with the control fruit,fruit softening was inhibited in slnac4 fruit and conversely was accelerated in OE-SlNAC4 tomato fruit.Through RNA-sequencing(RNA-seq)analysis,we found that expression levels of SlEXP1(expansin)and SlCEL2(endo-β-1,4 glucanase)genes involved in cell wall metabolism were significantly different in WT(wild type)/slnac4 and WT/OE-SlNAC4 fruit.Further study showed that these genes contained a NAC TF binding domain in their promoter regions.In vitro electrophoretic mobility shift assays(EMSA)and dual-luciferase reporter assays(DLR)demonstrated that these two genes were the direct targets of SlNAC4 binding and transactivation.The results enriched the function of SlNAC4 and provided a new dimension in understanding the regulation of tomato fruit softening.

A novel lectin from mushroom Phellodon melaleucus displays hemagglutination activity,and antitumor activity in a B16 melanoma mouse model

A novel lectin(termed PML)was purified from fruiting bodies of the edible mushroom Phellodon melaleucus(division Basidiomycota)by ion exchange,hydrophobic interaction,and gel filtration chromatographies,with overall titer recovery~60%and 20-fold purification.PML displayed hemagglutination activity 13319 units/mg toward rabbit erythrocytes.SDS-PAGE and gel filtration analyses revealed that PML is a homodimeric lectin with a molecular weight of 28.8 kDa.PML hemagglutination activity was not inhibited by various simple sugars or their derivatives,but was enhanced by cations Ca^(2+),Mg^(2+),Zn^(2+),and Cu^(2+).The activity was stable in pH range 6–9 and in the temperature range 20–60°C.Circular dichroism(CD)spectroscopic analysis showed that PML was composed primarily ofβ-sheets with lowα-helix content.In a B16 melanoma mouse model,PML treatment significantly inhibited tumor growth,and increased cytokine IL-10 content.Our findings suggest that PML is a potential anticancer therapeutic agent.

Another piece of puzzle for the human microbiome: the gut virome under dietary modulation

The virome is the most abundant and highly variable microbial consortium in the gut.Because of difficulties in isolating and culturing gut viruses and the lack of reference genomes,the virome has remained a rela-tively elusive aspect of the human microbiome.In recent years,studies on the virome have accumulated growing evidence showing that the virome is diet-modulated and widely involved in regulating health.Here,we review the responses of the gut virome to dietary intake and the potential health implications,presenting changes in the gut viral community and preferences of viral members to particular diets.We further discuss how viral-bacterial interactions and phage lifestyle shifts shape the gut microbiota.We also discuss the specific functions conferred by diet on the gut virome and bacterial community in the context of horizontal gene transfer,as well as the import of new viral members along with the diet.Collating these studies will expand our understanding of the dietary regulation of the gut virome and inspire dietary in-terventions and health maintenance strategies targeting the gut microbiota.

Chitin oligosaccharides alleviate atherosclerosis progress in ApoE;mice by regulating lipid metabolism and inhibiting inflammation

Atherosclerosis is driven both by hyperlipidemia and inflammation. Chitin oligosaccharides(NACOS) have shown pharmacological effects on multiple diseases via hypolipidemic and/or anti-inflammatory activities. The present study aims to evaluate whether NACOS treatment can prevent atherosclerosis induced by a highfat-diet(HFD) in Apolipoprotein E-knockout(Apo E;) mice. Results showed that 300 and 900 mg/kg b.w./day NACOS supplementation for 14 weeks significantly decreased atherosclerotic lesions up to 45% and 67% in compared with the HFD(P < 0.05), as measured in the valve area of the aortic root. Further, NACOS supplementation significantly reduced the serum hyperlipidemia and circulating proinflammatory cytokines including interleukin-1β, interleukin-6, monocyte chemoattractant protein-1 and tumor necrosis factor-α. NACOS decreased the hepatic Hmgcr to reduce cholesterol synthesis, activated the genes involved in reverse cholesterol transport to enhance cholesterol efflux and excretion, and reduced the intestinal Npc1L1 to lower cholesterol absorption. Additionally, NACOS enhanced cecum short chain fatty acids production and intestinal integrity. Thus, NACOS supplementation ameliorated atherosclerosis via altering lipid metabolism and reducing inflammation. These findings indicate that NACOS may be a potential functional food material for attenuating atherosclerosis development.

Gut microbiota from end-stage renal disease patients disrupt gut barrier function by excessive production of phenol

End stage renal disease (ESRD), an advanced stage of chronic kidney disease, has received increasing attention as a leading public health problem (Webster et al., 2017). Chronic kidney disease (CKD) and ESRD are invariably accompanied by chronic systemic inflammation which plays central roles in CKD progression (Cachofeiro et al., 2008) and the increased mortality due to cardiovascular disease (Landray et al., 2004).

Assessment and distinction of commercial soy protein isolate product functionalities using viscosity characteristic curves

To simplify the assessment method of soy protein isolate （SPI） functionalities, the viscosity and functionalities of commercial SPI products were studied. Viscosity value （y） increases With increasing concentration （x） and exhibits a highly significant correlation with the exponential equation y = a. ebx. The b values of products are gradually enhanced from dispersion, emulsion and injected to gel type. Products with low b values （〈0.2）, and high dispersivity were dispersion-type. Products having high b values （〉0.4） and gel springiness were gel-type. The other products with centered b value （0.2-0.4）, high solubility and emulsifying capacity were emulsion-type.

Applications of virus-induced gene silencing for identification of gene function in fruit

With the development of bioinformatics,it is easy to obtain information and data about thousands of genes,but the determi nation of the functions of these genes depends on methods for rapid and effective functi on al identification.Virus-induced gene sile ncing(VIGS)is a mature method of gene functional identification developed over the last 20 years,which has been widely used in many research fields involving many species.Fruit quality formation is a complex biological process,which is closely related to ripening.Here,we review the progress and contribution of VIGS to our understanding of fruit biology and its advantages and disadvantages in determining gene function.

Efficient production of poly-γ-glutamic acid by Bacillus velezensis via solid-state fermentation and its application

A novel glutamate-dependent poly-γ-glutamic acid(γ-PGA)-producing strain Bacillus velezensis CAU263 isolated from Chinese traditional Douchi was evaluated.An efficient method ofγ-PGA production was performed by this strain via solid-state fermentation(SSF)using guar meal.The maximal yield ofγ-PGA reached 158.5 g/kg dry weight(DW)in 250 mL flasks and 155.1 g/kg DW in shallow tray,respectively.The molecular weight ofγ-PGA was 3.8×106 Da,the ratio of L/D-glutamic acid was 26.1%and 73.9%.Additionally,γ-PGA significantly improved the operating characteristics of wheat dough,the specific volume of bread was increased by 15.8%and the bread hardness was reduced by 44.2%owing to the addition of 0.3 g/kgγ-PGA.Thus,γ-PGA could be produced by Bacillus velezensis CAU263 at high level using guar meal and might have potential application prospect in food industry.

Maternal and infant microbiome:next-generation indicators and targets for intergenerational health and nutrition care

Microbes are commonly sensitive to shifts in the physiological and pathological state of their hosts,including mothers and babies.From this perspective,the microbiome may be a good indicator for diseases during pregnancy and has the potential to be used for perinatal health monitoring.This is embodied in the application of microbiome from multi body sites for auxiliary diagnosis,early prediction,prolonged monitoring,and retrospective diagnosis of pregnancy and infant complications,as well as nutrition management and health products developments of mothers and babies.Here we summarized the progress in these areas and explained that the microbiome of different body sites is sensitive to different diseases and their microbial biomarkers may overlap between each other,thus we need to make a diagnosis prudently for those diseases.Based on the microbiome variances and additional anthropometric and physical data,individualized responses of mothers and neonates to meals and probiotics/prebiotics were predictable,which is of importance for precise nutrition and probiotics/prebiotics managements and developments.Although a great deal of encouraging performance was manifested in previous studies,the efficacy could be further improved by combining multi-aspect data such as multi-omics and time series analysis in the future.This review reconceptualizes maternal and infant health from a microbiome perspective,and the knowledge in it may inspire the development of new options for the prevention and treatment of adverse pregnancy outcomes and bring a leap forward in perinatal health care.

Virus-induced Gene Silencing in Eggplant(Solanum melongena)

Eggplant （Solanum melongena） is an economically important vegetable requiring investigation into its various genomic functions. The current limitation in the investigation of genomic function in eggplant is the lack of effective tools available for conducting functional assays. Virus-induced gene silencing （VIGS） has played a critical role in the functional genetic analyses. In this paper, TRV-mediated VIGS was successfully elicited in eggplant. We first cloned the CDS sequence of PDS （PHYTOENE DESATURASE） in eggplant and then silenced the PDS gene. Photo-bleaching was shown on the newly-developed leaves four weeks after agroinoculation, indicating that VIGS can be used to silence genes in eggplant. To further illustrate the reliability of VIGS in eggplant, we selected Chl H, Su and CLA1 as reporters to elicit VIGS using the high-pressure spray method. Suppression of Chl H and Su led to yellow leaves, while the depletion of CLA1 resulted in albino. In conclusion, four genes, PDS, Chl H, Su （Sulfur）, CLA1, were down-regulated significantly by VIGS, indicating that the VIGS system can be successfully applied in eggplant and is a reliable tool for the study of gene function.