Star1 gene mutation reveals the essentiality of 11-ketotestosterone and glucocorticoids for male fertility in Nile Tilapia (Oreochromis niloticus).

Steroidogenic acute regulatory protein (Star) plays an essential role in the biosynthesis of corticosteroids and sex steroids by mediating the transport of cholesterol from the outer to the inner membrane of mitochondria. Two duplicated Star genes, namely star1 and star2, have been identified in non-mammalian vertebrates. To investigate the roles of star genes in fish steriodogenesis, we generated two mutation lines of star1-/- and star1-/-/star2-/- in Nile tilapia (Oreochromis niloticus). Previous studies revealed that deficiency of star2 gene caused delayed spermatogenesis, sperm apoptosis and sterility in male tilapia. Our present data revealed that mutation of star genes impaired male fertility. Disordered seminiferous lobules and spermatic duct obstruction were found in the testis of both types of mutants. Moreover, significant decline in semen volume, sperm abnormality and impaired fertility were also detected in star1-/- and star1-/-/star2-/- males. In star1-/- male fish, lipid accumulation, up-regulation of steroidogenic enzymes, and significant decline of androgens were found. Additionally, hyperplasic interrenal cells, elevated steroidogenic gene expression level and decline of serum glucocorticoids were detected in star1 mutants. Intriguingly, either 11-KT or cortisol supplementation successfully rescued the impaired fertility of the star1-/- mutants. Taken together, these results further indicate that Star1 might play critical roles in the production of both 11-KT and glucocorticoids, which are indispensable for the maintenance of male fertility in fish.

Hypogonadotropic hypogonadism in male tilapia lacking a functional rln3b gene.

Relaxin 3 is a neuropeptide that plays a crucial role in reproductive functions of mammals. Previous studies have confirmed that rln3a plays an important role in the male reproduction of tilapia. To further understand the significance of its paralogous gene rln3b in male fertility, we generated a homozygous mutant line of rln3b in Nile tilapia. Our findings indicated that rln3b mutation delayed spermatogenesis and led to abnormal testes structure. Knocking out rln3b gene resulted in a decrease in sperm count, sperm motility and male fish fertility. TUNEL detection revealed a small amount of apoptosis in the testes of rln3b-/- male fish at 390 days after hatching (dah). RT-qPCR analysis demonstrated that mutation of rln3b gene caused a significant downregulation of steroid synthesis-related genes such as cyp17a1, cyp11b2, germ cell marker gene, Vasa, and gonadal somatic cell marker genes of amh and amhr2. Furthermore, we found a significant down-regulation of hypothalamic-pituitary-gonadal (HPG) axis-related genes, while a significantly up-regulation of the dopamine synthetase gene in the rln3b-/- male fish. Taken together, our data strongly suggested that Rln3b played a crucial role in the fertility of XY tilapia by regulating HPG axis genes.

Genome-wide analysis of two different regions of brain reveals the molecular changes of fertility related genes in rln3a-/- mutants in male Nile tilapia (Oreochromis niloticus).

Relaxin3 (rln3) has been associated with various emotional and cognitive processes, including stress, anxiety, learning, memory, motivational behavior, and circadian rhythm. Notably, previous report revealed that Rln3a played an indispensable role in testicular development and male fertility in Nile tilapia (Oreochromis niloticus). However, the underlying molecular mechanisms remain largely unknown. We found that Rln3a is expressed exclusively in the diencephalon* (Di*) of the brain. Deficiency of Rln3a resulted in a significant increase in serum dopamine level and an upregulation of gene expression of gnrh1 and kisspeptin2. To further elucidate the role of Rln3a in fish fertility, we collected two different regions of Di* and hypothalamus (Hyp) tissues for subsequent RNA-seq analysis of both wild-type (rln3a+/+) and rln3a-/- male tilapia. Upon the transcriptomic data, 1136 and 755 differentially expressed genes (DEGs) were identified in the Di* and Hyp tissues, respectively. In Di*, the up-regulated genes were enriched in circadian rhythm, chemical carcinogenesis, while the down-regulated genes were enriched in type II diabetes mellitus, dopaminergic synapse, and other pathways. In Hyp, the up-regulated genes were enriched in circadian rhythm, pyrimidine metabolism, while the down-regulated genes were enriched in type I diabetes mellitus, autoimmune thyroid disease, and other pathways. Subsequently, the results of both qRT-PCR and FISH assays highlighted a pronounced up-regulation of core circadian rhythm genes, cry1b and per3, whereas genes such as clocka, clockb, and arntl exhibited down-regulation. Furthermore, the genes associated with dopamine biosynthesis were significantly increased in the Hyp. In summary, the mutation of rln3a in male tilapia resulted in notable changes in circadian rhythm and disease-linked signaling pathways in the Di* and Hyp. These changes might account for the fertility defects observed in rln3a-/- male mutants in tilapia.

Exploring the Blood Glucose-Lowering Potential of the Umami Peptides LADW and EEAEGT Derived from Tuna Skeletal Myosin: Perspectives from α-Glucosidase Inhibition and Starch Interaction.

This study aimed to explore the potential of umami peptides for lowering blood glucose. Molecular docking results showed that the peptides LADW and EEAEGT bound to the active amino acid residues of α-glucosidase via hydrogen bonds and Van der Waals forces, a finding supported by an independent gradient model (IGM). Molecular dynamics (MD) simulations demonstrated that the peptides LADW and EEAEGT can decelerate the outward expansion of α-glucosidase and reduce amino acid fluctuations at the active site. In vitro findings indicated that the peptides LADW and EEAEGT showed potent inhibitory activity against α-glucosidase, with IC50 values of 4.40 ± 0.04 and 6.46 ± 0.22 mM, respectively. Furthermore, MD simulation and morphological observation results also revealed that LADW and EEAEGT alter starch structure and form weak interactions with starch through intermolecular hydrogen bonding, leading to the inhibition of starch hydrolysis. Peptides inhibit the ability of starch to produce reducing sugars after simulated gastrointestinal digestion, providing additional evidence of the inhibition of starch hydrolysis by the added peptides. Taken together, these findings suggest that consuming the umami peptides LADW and EEAEGT may alleviate postprandial blood glucose elevations via inhibiting α-glucosidase and starch hydrolysis.

Comparison of an Ultrasound-Assisted Aqueous Two-Phase System Extraction of Anthocyanins from Pomegranate Pomaces by Utilizing the Artificial Neural Network-Genetic Algorithm and Response Surface Methodology Models.

As a by-product of pomegranate processing, the recycling and reuse of pomegranate pomaces (PPs) were crucial to environmentally sustainable development. Ultrasound-assisted aqueous two-phase extraction (UA-ATPE) was applied to extract the anthocyanins (ACNs) from PPs in this study, and the central composite design response surface methodology (CCD-RSM) and artificial neural network-genetic algorithm (ANN-GA) models were utilized to optimize the extraction parameters and achieve the best yield. The results indicated that the ANN-GA model built for the ACN yield had a greater degree of fit and accuracy than the RSM model. The ideal model process parameters were optimized to have a liquid-solid ratio of 49.0 mL/g, an ethanol concentration of 28 g/100 g, an ultrasonic time of 27 min, and an ultrasonic power of 330 W, with a maximum value of 86.98% for the anticipated ACN yield. The experimental maximum value was 87.82%, which was within the 95% confidence interval. A total of six ACNs from PPs were identified by utilizing UHPLC-ESI-HRMS/MS, with the maximum content of cyanidin-3-O-glucoside being 57.01 ± 1.36 mg/g DW. Therefore, this study has positive significance for exploring the potential value of more by-products and obtaining good ecological and economic benefits in the future.

The profiles of free, esterified and insoluble-bound phenolics in peach juice after high pressure homogenization and evaluation of their antioxidant capacities, cytoprotective effect, and inhibitory effects on α-glucosidase and dipeptidyl peptidase-â £.

The phenolic profiles, antioxidant capacities, cytoprotective effect, and α-glucosidase and DPP-IV inhibitory capacity of free (FP), esterified (EP) and insoluble-bound (IBP) phenolic fractions in 'Lijiang snow' peach juice after high pressure homogenization (HPH) were investigated, and the molecular docking was used to explore the enzyme inhibition mechanism. HPH increased total phenolic and total flavonoid contents in three fractions without changing compositions. The IC50 of radicals scavenged by three fractions were all reduced by HPH. The best inhibition on intracellular ROS production were found for phenolic fractions after HPH at 300 MPa, with ROS levels ranged within 95.26-119.16 %. HPH at 300 MPa reduced the apoptosis rates of FP and EP by 16.52 % and 9.33 %, respectively. All phenolic fractions showed effective inhibition on α-glucosidase and DPP-IV by formation of hydrogen bonding and van der Waals forces. This study explored the feasibility of HPH to enhance the phenolics and bioactivity of peach juice.

Effects of juicing methods on the bioactive compounds and flavor quality of 'Black-seed' pomegranate from three producing areas.

BACKGROUND: Color, nutrients and flavor are the key characteristics of pomegranate juice, but they are susceptible to processing methods and raw materials. In this study, the effects of aril juicing and whole fruit juicing methods on the composition of 'Black-seed' pomegranate juice from three producing areas were studied, including physicochemical parameters, color attributes, organic acids, sugars, phenolic compounds, and volatile compounds. RESULTS: The whole fruit juicing method resulted in higher juice yields of pomegranate fruit with 69.01-72.59%, hue angle values were 5.95-6.45°, and the juice remained red. The highest level of citric acid (21.21 g L-1 ), total acids (24.78 g L-1 ), and total anthocyanin content (435.59 mg L-1 ) were found in whole fruit juice, and seven tannins were detected. The most abundant volatile compounds were (Z)-3-hexen-1-ol and 1-hexanol in all juice samples, with alcohol content increased and aldehydes content decreased by whole fruit juicing. Principal component analysis revealed that the 24 indexes (variable important in projection >1) clearly distinguished juice samples obtained by two juicing methods, with ellagic acid hexoside, (E)-2-heptenal, (+)-catechin, and octanoic acid having the best discriminatory potential. CONCLUSION: Overall, the effects of juicing method on 'Black-seed' pomegranate juice were greater than those of raw-material-producing areas. These results confirmed the potential for using the whole 'Black-seed' pomegranate for processing, and also provided a theoretical basis for the healthy product development and utilization of dark-color pomegranate varieties. © 2023 Society of Chemical Industry.

Endophytic Beauveria bassiana promotes plant biomass growth and suppresses pathogen damage by directional recruitment.

Introduction: Entomopathogenic fungi (EPF) can colonize and establish symbiotic relationships with plants as endophytes. Recently, EPF have been reported to suppress plant pathogens and induce plant resistance to diseases. However, the potential mechanisms via which EPF as endophytes control major plant diseases in situ remain largely unknown. Methods: Pot and field experiments were conducted to investigate the mechanisms via which an EPF, Beauveria bassiana, colonizes tomato, under Botrytis cinerea infection stress. B. bassiana blastospores were inoculated into tomato plants by root irrigation. Tomato resistance to tomato gray mold caused by B. cinerea was evaluated by artificial inoculation, and B. bassiana colonization in plants and rhizosphere soil under B. cinerea infection stress was evaluated by colony counting and quantitative PCR. Furthermore, the expression levels of three disease resistance-related genes (OXO, CHI, and atpA) in tomato leaves were determined to explore the effect of B. bassiana colonization on plant disease resistance performance in pot experiments. Results: B. bassiana colonization could improve resistance of tomato plants to gray mold caused by B. cinerea. The incidence rate, lesion diameter, and disease index of gray mold decreased in both the pot and field experiments following B. bassiana colonization. B. bassiana was more likely to accumulate in the pathogen infected leaves, while decreasing in the rhizosphere soil, and induced the expression of plant resistance genes, which were up-regulated in leaves. Discussion: The results indicated that plants could "recruit" B. bassiana from rhizosphere soil to diseased plants as directional effects, which then enhanced plant growth and resistance against pathogens, consequently inhibiting pathogen infection and multiplication in plants. Our findings provide novel insights that enhance our understanding of the roles of EPF during pathogen challenge.

Zebrafish gonad mutant models reveal neuroendocrine mechanisms of brain sexual dimorphism and male mating behaviors of different brain regions.

BACKGROUND: Sexually dimorphic mating behaviors differ between sexes and involve gonadal hormones and possibly sexually dimorphic gene expression in the brain. However, the associations among the brain, gonad, and sexual behavior in teleosts are still unclear. Here, we utilized germ cells-free tdrd12 knockout (KO) zebrafish, and steroid synthesis enzyme cyp17a1-deficient zebrafish to investigate the differences and interplays in the brain-gonad-behavior axis, and the molecular control of brain dimorphism and male mating behaviors. METHODS: Tdrd12+/-; cyp17a1+/- double heterozygous parents were crossed to obtain tdrd12-/-; cyp17a1+/+ (tdrd12 KO), tdrd12+/+; cyp17a1-/- (cyp17a1 KO), and tdrd12-/-; cyp17a1-/- (double KO) homozygous progenies. Comparative analysis of mating behaviors were evaluated using Viewpoint zebrafish tracking software and sexual traits were thoroughly characterized based on anatomical and histological experiments in these KOs and wild types. The steroid hormone levels (testosterone, 11-ketotestosterone and 17ß-estradiol) in the brains, gonads, and serum were measured using ELISA kits. To achieve a higher resolution view of the differences in region-specific expression patterns of the brain, the brains of these KOs, and control male and female fish were dissected into three regions: the forebrain, midbrain, and hindbrain for transcriptomic analysis. RESULTS: Qualitative analysis of mating behaviors demonstrated that tdrd12-/- fish behaved in the same manner as wild-type males to trigger oviposition behavior, while cyp17a1-/- and double knockout (KO) fish did not exhibit these behaviors. Based on the observation of sex characteristics, mating behaviors and hormone levels in these mutants, we found that the maintenance of secondary sex characteristics and male mating behavior did not depend on the presence of germ cells; rather, they depended mainly on the 11-ketotestosterone and testosterone levels secreted into the brain-gonad regulatory axis. RNA-seq analysis of different brain regions revealed that the brain transcript profile of tdrd12-/- fish was similar to that of wild-type males, especially in the forebrain and midbrain. However, the brain transcript profiles of cyp17a1-/- and double KO fish were distinct from those of wild-type males and were partially biased towards the expression pattern of the female brain. Our results revealed important candidate genes and signaling pathways, such as synaptic signaling/neurotransmission, MAPK signaling, and steroid hormone pathways, that shape brain dimorphism and modulate male mating behavior in zebrafish. CONCLUSIONS: Our results provide comprehensive analyses and new insights regarding the endogenous interactions in the brain-gonad-behavior axis. Moreover, this study revealed the crucial candidate genes and neural signaling pathways of different brain regions that are involved in modulating brain dimorphism and male mating behavior in zebrafish, which would significantly light up the understanding the neuroendocrine and molecular mechanisms modulating brain dimorphism and male mating behavior in zebrafish and other teleost fish.

Integration of Untargeted Metabolomics and Object-Oriented Data-Processing Protocols to Characterize Acerola Powder Composition as Functional Food Ingredient.

Acerola powder has been experiencing a surge in demand as a functional food ingredient, particularly due to its usage in vitamin C supplements. However, limited research has been conducted on its other bioactive compounds. In this study, we employed metabolomics and object-oriented data-processing protocols to comprehensively characterize acerola powder. To ensure maximum coverage of metabolomics, we selected a 50% methanol aqueous solution as the extraction solvent and utilized the HSS T3 column for chromatography analysis. Through this approach, we successfully identified a total of 175 compounds in acerola powder, encompassing amino acids and peptides, polyphenols, organic acids, and various other compounds. Additionally, we measured the total phenolic content (TPC) and assessed the antioxidant activity of acerola powder. Furthermore, we analyzed the differential composition of acerola fruit and juice powder, identifying polyphenols and lipids as primary markers in fruit powder, while peptides emerged as key markers in juice powder. Notably, two specific peptides, Thr-Trp and Val-Tyr, were identified as antioxidant peptides. Overall, our study provides novel composition data for acerola powder, shedding light on its potential as a functional food ingredient. These findings contribute to the development and utilization of acerola powder in the formulation of functional food products.

Comparing the Effect of HPP on the Structure and Stability of Soluble and Membrane-Bound Polyphenol Oxidase from 'Lijiang Snow' Peach: Multispectroscopic and Molecular Dynamics Simulation.

Polyphenol oxidase (PPO) easily causes fruits and vegetables to lose their color and nutritional value. As a non-thermal process, high-pressure processing (HPP) showed different inactivation effects on endogenous enzymes. In this work, soluble PPO (sPPO) and membrane-bound PPO (mPPO) from 'Lijiang snow' peaches were purified, and then the effect of high pressure on the conformation of sPPO and mPPO was investigated and compared at the molecular level. The maximum activation of sPPO and mPPO by 11.2% and 4.8% was observed after HPP at 200 MPa, while their activities both gradually decreased at 400 MPa and 600 MPa; in particular, the residual activities of sPPO and mPPO at 600 MPa for 50 min were 41.42% and 72.95%, respectively. The spectroscopic results indicated that the secondary structure of PPOs was little affected by HPP, but HPP led to obvious changes in their tertiary structure. The simulations showed that the decreasing distance between the copper ion and His residue in the copper-binding region of two PPOs at 200 MPa was favorable to catalytic activity, while the increasing distance between copper ions and His residues and the disordered movement of the loop region above 400 MPa were unfavorable. In addition, the structure of sPPO was relatively looser than that of mPPO, and high pressure showed a more significant effect on the conformation of sPPO than that of mPPO. This study clarified the effect of HPP on PPO's structure and the relationship between its structure and activity and provided a basis for the prevention of enzymatic browning.

Fabrication of anthocyanin-rich W1/O/W2 emulsion gels based on pectin-GDL complexes: 3D printing performance.

The stability of anthocyanin-rich W1/O/W2 double emulsions prepared with Nicandra physalodes (Linn.) Gaertn. Seeds pectin was investigated, including droplet sizes, ζ-potential, viscosity, color, microstructures and encapsulation efficiency. Furthermore, the gelation behavior, rheological behavior, texture behavior and three-dimensional (3D) printing effects of the W1/O/W2 emulsion gels induced with Glucono-delta-lactone (GDL) were studied. The L*, b*, ΔE, droplet sizes and ζ-potential of the emulsions were gradually increased, while other indicators were gradually decreased during 28 days of storage under 4 ℃. The storage stability of sample under storage at 4 ℃ was higher than 25 ℃. The G' of W1/O/W2 emulsion gels gradually boosted with increased GDL addition, and reached the highest after the addition of 1.6 % GDL. In creep-recovery sweep, the minimum strain of 1.68 % and the highest recovery rate of 86 % were also found for the emulsion gels with 1.6 % GDL. Accordingly, the models "KUST", hearts, flowers printed by emulsion gels after 60 min addition of 1.6 % GDL had the best printing effects. The W1/O/W2 emulsion gels based on pectin-GDL complexes exhibited good performance in protecting anthocyanins and suggested as a potential ink for food 3D printing.

Gelation behavior and mechanism of Nicandra physalodes (Linn.) Gaertn. seeds pectin induced by Glucono-delta-lactone.

In this study, the physicochemical properties of pectin from Nicandra physalodes (Linn.) Gaertn. seeds (NPGSP) were analysed firstly, and the rheological behavior, microstructure and gelation mechanism of NPGSP gels induced by Glucono-delta-lactone (GDL) were investigated. The hardness of NPGSP gels was increased from 26.27 g to 226.77 g when increasing GDL concentration from 0 % (pH = 4.0) to 1.35 % (pH = 3.0), and the thermal stability was improved. The peak around 1617 cm-1 was decreased as the adsorption peak of the free carboxyl groups was attenuated with addition of GDL. GDL increased the crystalline degree of NPGSP gels, and its microstructure exhibited more smaller spores. Molecular dynamics was performed on systems of pectin and gluconic acid (GDL hydrolysis product), indicating that inter-molecular hydrogen bonds and van der Waals forces were the main interactions to promote gels formation. Overall, NPGSP has the potential commercial value for developing as a thickener in food processing.

Secrets behind Protein Sequences: Unveiling the Potential Reasons for Varying Allergenicity Caused by Caseins from Cows, Goats, Camels, and Mares Based on Bioinformatics Analyses.

This study systematically investigated the differences in allergenicity of casein in cow milk (CM), goat milk (GM), camel milk (CAM), and mare milk (MM) from protein structures using bioinformatics. Primary structure sequence analysis reveals high sequence similarity between the α-casein of CM and GM, while all allergenic subtypes are likely to have good hydrophilicity and thermal stability. By analyzing linear B-cell epitope, T-cell epitope, and allergenic peptides, the strongest casein allergenicity is observed for CM, followed by GM, and the casein of MM has the weakest allergenicity. Meanwhile, 7, 9, and 16 similar or identical amino acid fragments in linear B-cell epitopes, T-cell epitopes, and allergenic peptides, respectively, were observed in different milks. Among these, the same T-cell epitope FLGAEVQNQ was shared by κ-CN in all four different species' milk. Epitope results may provide targets of allergenic fragments for reducing milk allergenicity through physical or/and chemical methods. This study explained the underlying secrets for the high allergenicity of CM to some extent from the perspective of casein and provided new insights for the dairy industry to reduce milk allergy. Furthermore, it provides a new idea and method for comparing the allergenicity of homologous proteins from different species.

Nile tilapia (Oreochromis niloticus) Nanog co-expression with Pou5f3, transcriptional regulation and biological activity in embyonic development and embryonic cells.

Mammalian Nanog is critical in pluripotency acquisition and maintenance. Nonetheless, a recent report from zebrafish (Danio rerio) suggests that Nanog is not required for embryonic cells which is not like the mammalian homologs, but is necessary for the proper formation of the extra-embryonic yolk syncytial layer (YSL). However, whether its biological function in other fishes is conservative remains to be investigated. Our previous work shows that Nanog from Nile tilapia (Oreochromis niloticus) (termed as Ong thereafter) displays differential spatiotemporal expression patterns from the other teleost fishes including zebrafish. In this study, Ong co-expression with Pou5f3 (another core pluripotent transcription factor), transcriptional regulation and its biological functions during embryonic development and in the survival and proliferation of embryonic cells were investigated. At the blastula stage, both Ong and Pou5f3 were highly expressed in embryonic cells and co-located in the nucleus. After that, the expression of both Ong and Pou5f3 began to decrease at the gastrula stage (24 haf) and then exhibited a differential expression profile at the segmentation stage (28-36 haf). Ong disappeared in embryonic cells and was limited to YSL, whilst Pou5f3 was highly expressed in embryonic cells even some with obvious cytoplasmic distribution. Luciferase assay indicated that Ong was negatively regulated by Pou5f3 and positively regulated by androgen and itself. Ong depletion in fertilized one-cell embryos through CRISPR/Cas9 led to blastula blockage or death, and the survival and proliferation of blastula-derived embryonic cells in vitro failed. Collectively, Ong has similar expression and biological function to Pou5f3 at the blastula stage, which is similar to mammalian homolog but different from zebrafish homolog. These data suggest that the expression patterns and functions of Nanog are not conservative in fishes and vary from species to species. This study enriches our understanding about Nanog and its evolution.

Characterization of the flavor, sensory quality and in vitro bioaccessibility in cloudy pomegranate juice treated by high pressure and thermal processing.

BACKGROUND: Recently, cloudy pomegranate juice (PJ) has become popular due to its rich phenolic and health-promoting effects. The aim of the present work was to evaluate the application of high hydrostatic pressure processing (HPP), pasteurization (PT) and high-temperature short-time sterilization (HTST) on physicochemical properties (color, flow behavior, turbidity, sugars, organic acids, aroma and sensory evaluation) and in vitro bioaccessibility of total phenolics content (TPC), total flavonoids content (TFC) and phenolics of cloudy PJ. RESULTS: Compared to HPP, thermal sterilization significantly increased the brightness (L*), redness (a*), total color difference (ΔE) and turbidity, and decreased the TPC and TFC. HPP maintained the volatile profile of cloudy PJ better, while thermal sterilization significantly changed the profile by decreasing alcohols 23.8-32.7% and increasing acids by 33.6%-182.8%. The bioaccessibility of flavonoids, phenolic acids and tannins in the control cloudy PJ after in vitro oral-gastric-intestinal digestion were 1.5%, 4.9%, and 9.0%, respectively, which were not significantly changed by different treatments. CONCLUSION: These results contributed to promoting the color quality and health benefits of cloudy PJ rich in phenolics by optimizing the processing conditions in the food industry. © 2022 Society of Chemical Industry.

Comparison on inhibitory effect and mechanism of inhibitors on sPPO and mPPO purified from 'Lijiang snow' peach by combining multispectroscopic analysis, molecular docking and molecular dynamics simulation.

The inhibition mechanisms of soluble PPO (sPPO) by l-cysteine, reduced glutathione and thiourea, and membrane-bound (mPPO) by l-cysteine, reduced glutathione, thiourea, anisaldehyde and cinnamaldehyde were investigated by combining multispectroscopic analysis and computational simulations. Reduced glutathione showed the strongest inhibitory effect, with IC50 of 0.46 and 0.94 mM, respectively. The multispectral results showed that all inhibitors inhibited activity by destroying the secondary and tertiary structure, and the structure of sPPO were more easily affected. Docking showed that hydrogen bond and metal contact were the main driving force for inhibitors binding to sPPO and mPPO, respectively. Simulation showed that sPPO-inhibitor system had more fluctuation than mPPO-inhibitor system, indicating easier inhibition of sPPO activity. This work revealed that the structural differences between sPPO and mPPO led to different inhibition mechanisms of PPOs by inhibitors at the molecular level, which could provide the guidance for the selection of inhibitors in fruit and vegetable processing.

Cyp17a2 is involved in testicular development and fertility in male Nile tilapia, Oreochromis niloticus.

Background: Steroid hormones play an essential role in many reproductive processes of vertebrates. Previous studies revealed that teleost-specific Cyp17a2 (cytochrome P450 family 17 subfamily a 2) might be required for the production of cortisol in the head-kidney and 17α,20ß-dihydroxy-4-pregnen-3-one (DHP) in ovary during oocyte maturation. However, the role of Cyp17a2 in male reproduction remains to be largely unknown. The aim of this study was to explore the essentiality of cyp17a2 gene in male steroidogenesis, spermatogenesis, and male fertility. Methods: A homozygous mutation line of cyp17a2 gene was constructed in tilapia by CRISPR/Cas9 gene editing technology. The expression level of germ cell and meiosis-related genes and steroidogenic enzymes were detected by qRT-PCR, IHC, and Western blotting. EIA and LC-MS/MS assays were used to measure the steroid production levels. And sperm quality was examined by Sperm Quality Analyzer software. Results: In this study, cyp17a2 gene mutation resulted in the significant decline of serum DHP and cortisol levels. On the contrary, significant increases in intermediate products of cortisol and DHP were found in cyp17a2-/- male fish. The deficiency of cyp17a2 led to the arrest of meiotic initiation in male fish revealing as the reduction of the expression of germ cell-related genes (vasa, piwil, oct4) and meiosis-related genes (spo11 and sycp3) by 90 dah. Afterwards, spermatogenesis was gradually recovered with the development of testis in cyp17a2-/- males, but it showed a lower sperm motility and reduced fertility compared to cyp17a2+/+ XY fish. Deletion of cyp17a2 led to the abnormal upregulation of steroidogenic enzymes for cortisol production in the head-kidney. Moreover, unaltered serum androgens and estrogens, as well as unchanged related steroidogenic enzymes were found in the testis of cyp17a2-/- male fish. Conclusion: This study proved that, for the fist time, Cyp17a2 is indispensable for cortisol and DHP production, and cyp17a2 deficiency associated curtailed meiotic initiation and subfertility suggesting the essentiality of DHP and cortisol in the male fertility of fish.

Chestnut Starch Nanocrystal Combined with Macadamia Protein Isolate to Stabilize Pickering Emulsions with Different Oils.

This study investigated the formation and molecular interaction mechanism of chestnut starch nanocrystal (SNC)/macadamia protein isolate (MPI) complexes and their application in edible oil-in-water Pickering emulsion (PE). SNC/MPI complexes were characterized by scanning electron microscopy and particle size analyzer. The PEs stabilized by SNC/MPI complexes were characterized by confocal laser scanning microscopy and rheological measurement. The results showed that hydrogen bonds between the two particles significantly affected the secondary structure and assembly of SNC/MPI complexes at the oil/water interface. The optimal mass ratio of SNC to MPI in the complexes with the best stability was determined as 20:1. The formation of edible oil-in-water PEs stabilized by SNC/MPI complexes significantly improved the oxidative and storage stability of different edible oils (olive oil, walnut oil, edible tea oil, and macadamia oil). These different edible oil-in-water PEs stabilized by SNC/MPI could be used as effective carriers of quercetin with their loading rates higher than 93%.

Water Caltrop (Trapa quadrispinosa Roxb.) Husk Improves Oxidative Stress and Postprandial Blood Glucose in Diabetes: Phenolic Profiles, Antioxidant Activities and α-Glycosidase Inhibition of Different Fractions with In Vitro and In Silico Analyses.

The aim of this study was to investigate the phenolic profiles, antioxidant activities and α-glycosidase inhibitory activities of three different phenolic fractions from water caltrop (Trapa quadrispinosa Roxb.) husk and to further explore the predominant compounds and their mechanisms on α-glycosidase inhibition by virtual screening and molecular dynamics. A total of 29 substances were identified and quantified in this study. Tannins were the main constituents of water caltrop husk extract. All of the free phenolic (FP), esterified phenolic (EP) and insoluble-bound phenolic (BP) fractions exhibited good antioxidant activities, and the BP had the highest radical scavenging ability with IC50 values of 0.82 ± 0.12 µg/mL (ABTS) and 1.15 ± 0.02 µg/mL (DPPH), respectively (p < 0.05). However, compared with the EP and BP, the FP showed the strongest inhibition towards the α-glycosidase and the IC50 value of FP was 1.43 ± 0.12 µg/mL. The 1,2,6-trigalloylglucose and α-glycosidase complex had better root mean square deviations (RMSD) stability via molecular dynamics simulation study. Results obtained from this study may provide a good potential natural resource for the improvement of oxidative stress injury and blood glucose control in diabetes mellitus, which could expand the use of water caltrop husk and improve its economic value.