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Objective: To obtain the optimal enzyme and preparation process for Schisandra chinensis protein peptides and investigate its in vitro antioxidant activity. Methods: Seven proteases were used to hydrolyze Schisandra chinensis protein. Based on the degree of hydrolysis, free radical scavenging activity, polypeptide yield and content of different hydrolysates of Schisandra chinensis protein and the comprehensive evaluation of molecular weight in SDS-PAGE, the optimal protease was screened. The DPPH free radical scavenging rate was used as the index, and the optimal enzymatic hydrolysis process was determined by single factor test combined with response surface analysis. The scavenging ability of O2−·, ·OH, DPPH·, ABTS+·, Fe2+ chelating ability and Fe3+ reducing ability of Schisandrae chinensis protein peptides were analyzed and compared with Schisandra chinensis protein. Results: The optimum enzyme for the preparation of Schisandra chinensis protein peptides was alkaline protease. The optimum enzymatic hydrolysis parameters were as follows: substrate concentration 5%, enzyme-to-substrate ratio 1%, enzymatic hydrolysis time 3 h, enzymatic hydrolysis temperature 55 ℃, pH9.0. Under these conditions, the polypeptide content was 88.61%, the degree of hydrolysis was 24.21%, and the DPPH· scavenging rate was 86.96%. The free radical scavenging ability and reducing ability of Schisandrae chinensis protein peptides were better than those of Schisandrae chinensis protein. Conclusion: This study determined the optimum enzyme and hydrolysis process of Schisandrae chinensis protein peptides, and pointed out that Schisandrae chinensis protein peptides had better antioxidant activity in vitro and could be used as a natural antioxidant.

Red grapes often suffer from postharvest diseases like blue mold and black mold caused by Penicillium expansum and Aspergillus niger. Biological control using beneficial yeasts and bacteria is an effective method to manage these diseases. Rhodotorula sp. and Bacillus sp. are effective microorganisms for the control of postharvest diseases of red grapes. This study combined two yeast strains (Rhodotorula graminis and Rhodotorula babjevae) and two bacterial strains (Bacillus licheniformis and Bacillus velezensis) to investigate their biological control effects on major postharvest diseases of red grapes and explore the underlying physiological mechanisms. Research showed that compound microorganism W3 outperformed the others; it reduced spore germination and germ tube growth of P. expansum and A. niger, while its volatiles further inhibited pathogen growth. Additionally, the treatment enhanced the antioxidant capacity of grapes and increased resistance to pathogens by boosting peroxidase activities, superoxide dismutase, catalase and ascorbate peroxidase, phenylalanine ammonolyase, and polyphenol oxidase. Furthermore, the combined treatment increased the activity and accumulation of antifungal compounds such as total phenols and flavonoids, thereby improving disease resistance and reducing decay. Therefore, composite microorganisms combining various antagonistic strains may offer a viable substitute for tackling postharvest diseases in red grapes.

A wideband wide-scan array based on dual-polarized elements is presented in this paper. The proposed element consists of two flared metallic wings and two shorting posts over a ground plane which is excited by a RF connector vertically. Without complex feeding network, the antenna element features wideband, low-profile, and high power handling capability. Besides, elements do not require electrical connection and can be arranged with gaps between neighbors, allowing for modular construction. The dual-polarized array in a square lattice can achieve wideband wide-scan performance. To verify the design, an $11\times 11$ prototype has been fabricated and measured. The measurements of this prototype are presented and are in good agreement with the simulations. The results reveal that the array element can achieve a relative bandwidth of 100% for VSWR $\le 3$ at scans out to $\theta \,\, = \,\, 45^{\circ }$ and an enhanced gain of 1 dB in lower frequency band. The array can obtain better than 20 dB polarization purity at 4 GHz and scan loss of lower than 3 dB over the band, respectively.

BackgroundIsoniazid-resistant, rifampicin-susceptible tuberculosis (Hr-TB) globally exhibits a high prevalence and serves as a potential precursor to multidrug-resistant tuberculosis (MDR-TB). Recognizing the spatial distribution of Hr-TB and identifying associated factors can provide strategic entry points for interventions aimed at early detection of Hr-TB and prevention of its progression to MDR-TB. This study aims to analyze spatial patterns and identify socioeconomic, demographic, and healthcare factors associated with Hr-TB in Shanghai at the county level.MethodWe conducted a retrospective study utilizing data from TB patients with available Drug Susceptible Test (DST) results in Shanghai from 2010 to 2016. Spatial autocorrelation was explored using Global Moran’s I and Getis-Ord Gi∗ statistics. A Bayesian hierarchical model with spatial effects was developed using the INLA package in R software to identify potential factors associated with Hr-TB at the county level.ResultsA total of 8,865 TB patients with DST were included in this analysis. Among 758 Hr-TB patients, 622 (82.06%) were new cases without any previous treatment history. The drug-resistant rate of Hr-TB among new TB cases in Shanghai stood at 7.20% (622/8014), while for previously treated cases, the rate was 15.98% (136/851). Hotspot areas of Hr-TB were predominantly situated in southwestern Shanghai. Factors positively associated with Hr-TB included the percentage of older adult individuals (RR = 3.93, 95% Crl:1.93–8.03), the percentage of internal migrants (RR = 1.35, 95% Crl:1.15–1.35), and the number of healthcare institutions per 100 population (RR = 1.17, 95% Crl:1.02–1.34).ConclusionWe observed a spatial heterogeneity of Hr-TB in Shanghai, with hotspots in the Songjiang and Minhang districts. Based on the results of the models, the internal migrant population and older adult individuals in Shanghai may be contributing factors to the emergence of areas with high Hr-TB notification rates. Given these insights, we advocate for targeted interventions, especially in identified high-risk hotspots and high-risk areas.

Abstract Semen Ziziphi Spinosae protein (SZSP) is a new plant protein resource with good food functional properties and health care function. However, the biological activity of SZSP has not been further studied, which greatly limits the development and utilization of SZSP in the food industry. The aim of this study was to investigate the protective effect of SZSP on immunosuppressed mice and its inhibitory effect on immune-stimulated RAW264.7 cells. The results demonstrated that SZSP remarkably improved the immunomodulatory secretion in serum (interleukin-2, tumor necrosis factor-α [TNF-α], interferon-γ, immunoglobulin-A, immunoglobulin-G, immunoglobulin-M) and primary macrophages (nitric oxide, interleukin-1β, TNF-α) and promoted the NK-cell killing activity of primary splenocytes in CTX-induced immunosuppression mice. Immunohistochemical analysis results indicated that the secretion of CD4 + and CD8 + in the spleen and thymus can be regulated by SZSP, leading to inhibition of the damage induced by cyclophosphamide in mice. Meanwhile, in order to clarify the immunomodulatory mechanism of SZSP, we showed that SZSP significantly inhibited the secretion of NO, interleukin-6, and TNF-α and reduced the phosphorylation expression of p-ERK, p-JNK, and p-IκBα in lipopolysaccharide-stimulated RAW264.7 cells. Therefore, the immunomodulatory effect of SZSP may be related to the activation of MAPKs and NF-κB signaling pathways. Based on the above studies, the preliminary purification of SZSP was continued, and S1F2G1 with immunomodulatory activity was obtained. Taken together, SZSP has an immunoregulatory effect in vivo and in vitro and may be a favorable candidate of functional food raw material for regulating immune responses.

Postharvest diseases, such as black spots caused by Alternaria alternata, have caused huge economic losses to the tomato industry and seriously restricted its development. In recent years, biological control has become a new method to control postharvest diseases of fruits and vegetables. Our research group screened W. anomalus, a yeast demonstrating a promising control effect on a postharvest black spot disease of tomatoes, and explored its physiological mechanism of prevention and control. Therefore, this study investigated the prevention and control effect of metabolites of W. anomalus on tomato black spot disease and the inhibition effect of main components on A. alternata. A GC-MS analysis found that isoamyl acetate was the main component of W. anomalus that played an inhibitory role. The results showed that isoamyl acetate could inhibit the growth of A. alternata and had a certain control effect on postharvest black spots in tomatoes. Our findings suggest that isoamyl acetate could be a promising alternative to fungicides for controlling postharvest black spots in tomatoes.

BackgroundsThe diagnostic delay of tuberculosis (TB) contributes to further transmission and impedes the implementation of the End TB Strategy. Therefore, we aimed to describe the characteristics of patient delay, health system delay, and total delay among TB patients in Shanghai, identify areas at high risk for delay, and explore the potential factors of long delay at individual and spatial levels.MethodThe study included TB patients among migrants and residents in Shanghai between January 2010 and December 2018. Patient and health system delays exceeding 14 days and total delays exceeding 28 days were defined as long delays. Time trends of long delays were evaluated by Joinpoint regression. Multivariable logistic regression analysis was employed to analyze influencing factors of long delays. Spatial analysis of delays was conducted using ArcGIS, and the hierarchical Bayesian spatial model was utilized to explore associated spatial factors.ResultsOverall, 61,050 TB patients were notified during the study period. Median patient, health system, and total delays were 12 days (IQR: 3–26), 9 days (IQR: 4–18), and 27 days (IQR: 15–43), respectively. Migrants, females, older adults, symptomatic visits to TB-designated facilities, and pathogen-positive were associated with longer patient delays, while pathogen-negative, active case findings and symptomatic visits to non-TB-designated facilities were associated with long health system delays (LHD). Spatial analysis revealed Chongming Island was a hotspot for patient delay, while western areas of Shanghai, with a high proportion of internal migrants and industrial parks, were at high risk for LHD. The application of rapid molecular diagnostic methods was associated with reduced health system delays.ConclusionDespite a relatively shorter diagnostic delay of TB than in the other regions in China, there was vital social-demographic and spatial heterogeneity in the occurrence of long delays in Shanghai. While the active case finding and rapid molecular diagnosis reduced the delay, novel targeted interventions are still required to address the challenges of TB diagnosis among both migrants and residents in this urban setting.

Objective: To optimize the enzymatic hydrolysis process of Pueraria protein and study its antioxidant properties. Methods: DPPH radical scavenging ability and DH were used as the index to screen the best hydrolytic protease. Base on the single factor experiments, the Box-Behnken response surface design was used to optimize the enzymatic hydrolysis process of Pueraria protein, and the antioxidant properties of the optimum hydrolysate in vitro was analyzed. Results: The optimal hydrolysis conditions for alkaline protease were as follows: Temperature 55 ℃, pH9 and enzyme substrate ratio 2%. Under these conditions, the IC50 of DPPH·, ABTS+·, and ·OH radicals were 0.15, 0.38, 1.41 mg/mL and a reduction capacity of 0.553. Conclusion: Under these conditions, Pueraria protein hydrolysate had remarkable antioxidant properties.

The ability to learn continuously is crucial for a robot to achieve a high level of intelligence and autonomy. In this paper, we consider continual reinforcement learning (RL) for quadruped robots, which includes the ability to continuously learn sub-sequential tasks (plasticity) and maintain performance on previous tasks (stability). The policy obtained by the proposed method enables robots to learn multiple tasks sequentially, while overcoming both catastrophic forgetting and loss of plasticity. At the same time, it achieves the above goals with as little modification to the original RL learning process as possible. The proposed method uses the Piggyback algorithm to select protected parameters for each task, and reinitializes the unused parameters to increase plasticity. Meanwhile, we encourage the policy network exploring by encouraging the entropy of the soft network of the policy network. Our experiments show that traditional continual learning algorithms cannot perform well on robot locomotion problems, and our algorithm is more stable and less disruptive to the RL training progress. Several robot locomotion experiments validate the effectiveness of our method.

Cladosporium cladosporioides is an emerging pathogenic fungus that causes Cladosporium rot in postharvest table grapes (Vitis vinifera). However, studies investigating the infection process of C. cladosporioides are lacking. Therefore, this study aimed to elucidate the infection process by investigating the influence of microclimatic parameters (temperature, wetness and fungal age) in C. cladosporioides pathogenesis, activities of grape defense-related enzymes and grape quality during the infection. C. cladosporioides effectively infects grapes by developing distinct colonies on the artificial wounds and berry surfaces, completing its life cycle within 48 h. The C. cladosporioides disease incidence optimally occurred at 20 °C and 25 °C. Wetness played an influential role in the infectivity of C. cladosporioides and 7-day-old C. cladosporioides resulted in the most serious disease incidence of table grapes. As a result of infection, the quality of grapes was affected, including berry weight, pH, titratable acidity (TA), total soluble solids (TSS), and ascorbic acid level. This infection also induced defense-related enzymes, including polyphenoloxidase (PPO), peroxidase (POD), phenylalanine ammonialyase (PAL), and catalase (CAT), at certain times. The findings of this study demonstrated that Cladosporium rot development depended on the microclimatic parameters of grapes, significantly affected the grape quality and induced grape's defense-related enzymes.

Abstract Background Atopic dermatitis (AD) is a chronic inflammatory skin disease accompanied with itchy and scaly rash. Compound traditional Chinese medicine dermatitis ointment (CTCMDO) consists of a mixture of extracts from five plants, which had been used in AD treatment due to good anti-inflammatory and anti-allergic effects. Materials and methods In this study, high-performance liquid chromatography (HPLC) and liquid chromatography/mass spectrometer (LC/MS) were performed to analyze the active ingredients of CTCMDO in detail and to establish its HPLC fingerprint. Furthermore, the anti-inflammatory and antipruritic activities of CTCMDO were studied in the treatment of DNCB-induced AD in mice. Results A total of 44 compounds including phenylpropionic acid compounds, alkaloid compounds, curcumin compounds and lignans were identified via combined HPLC and LC/MS. A fingerprint with 17 common peaks was established. In AD-like mice, DNCB-induced scratching behavior had been suppressed in the treatment of CTCMDO in a dose-dependent manner. Furthermore, the detailed experimental results indicated that the AD can be effectively improved via inhibiting the production of Th1/2 cytokines in serum, reversing the upregulation of substance P levels of itch-related genes in the skin, and suppressing the phosphorylation of JNK, ERK, and p38 in the skin. Conclusion This work indicated that CTCMDO can significantly improve AD via attenuating the pathological alterations of Th1/2 cytokines and itch-related mediators, as well as inhibiting the phosphorylation of mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB).

Mechanical stress induced by compression during preharvest and postharvest handling of fresh fruit is a major factor influencing the quality of fresh fruit. The degree of compression primarily governs the extent of quality deterioration. However, research on the damage mechanisms of mechanical compression in table grapes remains scarce. This investigation evaluated the impact of varying compression levels (0%, 20%, 40%, 60%, and 80%) on the postharvest quality attributes of table grapes. Changes in postharvest physical properties (overall appearance and color), structural properties (firmness, springiness, cohesiveness, and chewiness), physiological qualities (total soluble solids, titratable acidity, ascorbic acid, malonaldehyde content, and relative electrical conductivity), and cell microstructure of the berries was determined at 4 °C during 15 d of storage. Mechanical compression contributed to the deterioration of the quality of table grapes during storage, resulting in increased weight loss, decay rate, malonaldehyde content, and relative electrical conductivity; and decreased total soluble solids, titratable acidity, and ascorbic acid content. Furthermore, as compared to the control group, mechanical compression resulted in substantial yellowing and diminished textural qualities of grapes. In particular, compression treatment caused significant deformation of grape cell microstructure. In conclusion, mechanical compression stress significantly affects the physical and physiological properties of postharvest table grapes, as well as the internal cellular organization. As compression levels increase, the quality of table grapes progressively deteriorates, leading to a substantial reduction in storage life and commercial value. This study offers essential information for devising damage prevention strategies in preharvest and postharvest handling of table grapes.

Longitudinal-torsional composite ultrasonic vibration has been widely used in grinding. This paper aims to solve the problem that the resonance frequency deviates greatly from the theoretical design frequency and the vibration mode is poor when the horn is matched with a larger tool head. This paper presents how the longitudinal-torsional composite ultrasonic conical transition horn was designed and optimized by the transfer matrix theory and finite element simulation. For this purpose, the spiral groove parameters were optimized and selected by finite element simulation. Then, the modal analysis and transient dynamic analysis of the horn with grinding wheel were carried out to verify the correctness of the theoretical calculation. The impedance analysis and amplitude test of the horn with grinding wheel were carried out. The test results were in very good agreement with the theoretical and simulation results. Finally, the grinding experiment was carried out. The surface roughness of the workpiece in longitudinal-torsional ultrasonic vibration grinding was obviously reduced compared to that of ordinary grinding. All these obtained results demonstrate that the designed longitudinal-torsional composite ultrasonic horn has very good operational performance for practical applications.

The genus Acorus, a perennial monocotyledonous-class herb and part of the Acoraceae family, is widely distributed in the temperate and subtropical zones of the Northern and Southern Hemispheres. Acorus is rich in biological activities and can be used to treat various diseases of the nervous system, cardiovascular system, and digestive system, including Alzheimer’s disease, depression, epilepsy, hyperlipidemia, and indigestion. Recently, it has been widely used to improve eutrophic water and control heavy-metal-polluted water. Thus far, only three species of Acorus have been reported in terms of chemical components and pharmacological activities. Previously published reviews have not further distinguished or comprehensively expounded the chemical components and pharmacological activities of Acorus plants. By carrying out a literature search, we collected documents closely related to Acorus published from 1956 to 2022. We then performed a comprehensive and systematic review of the genus Acorus from different perspectives, including botanical aspects, ethnic applications, phytochemistry aspects, and pharmacological aspects. Our aim was to provide a basis for further research and the development of new concepts.

β-1,3-glucanase plays an important role in the biodegradation, reconstruction, and development of β-1,3-glucan. An endo-β-1,3-glucanase which was encoded by PeBgl1 was expressed, purified and characterized from Penicillium expansum for the first time. The PeBgl1 gene was amplified and transformed into the competent cells of E. coli Rosetta strain with the help of the pET-30a cloning vector. The recombinant protein PeBgl1 was expressed successfully at the induction conditions of 0.8 mmol/L IPTG at 16 °C for 16 h and then was purified by nickel ion affinity chromatography. The optimum reaction temperature of PeBgl1 was 55 °C and it had maximal activity at pH 6.0 according to the enzymatic analysis. Na2HPO4-NaH2PO4 buffer (pH 6.0) and NaCl have inhibitory and enhancing effects on the enzyme activities, respectively. SDS, TritonX-100 and some metal ions (Mg2+, Ca2+, Ba2+, Cu2+, and Zn2+) have an inhibitory effect on the enzyme activity. The results showed that PeBgl1 protein has good enzyme activity at 50–60 °C and at pH 5.0–9.0, and it is not a metal dependent enzyme, which makes it robust for storage and transportation, ultimately holding great promise in green biotechnology and biorefining.

Ziziphus jujuba var. spinosa (Bunge) Hu ex H.F.Chow [Rhamnaceae; Ziziphi Spinosae Semen (ZSS)] has attracted extensive attention as the first choice of traditional Chinese medicine in the treatment of insomnia. However, recent studies on the sleep-improving mechanism of ZSS have mainly focused on the role of single components. Thus, to further reveal the potential mechanism of ZSS, an assessment of its multiple constituents is necessary. In this study, ZSS extract (ZSSE) was obtained from ZSS via detailed modern extraction, separation, and purification technologies. The chemical constituents of ZSSE were analyzed by high-performance liquid chromatography–mass spectrometry (HPLC–MS). For in vivo experiments, a rat model of insomnia induced by p-chlorophenylalanine (PCPA) was established to investigate the potential effect and corresponding mechanism of ZSSE on improving sleep. Hematoxylin–eosin staining (HE) results revealed that the drug group showed prominent advantages over the model group in improving sleep. Moreover, the brain levels of γ-aminobutyric acid (GABA), glutamic acid (Glu), 5-hydroxytryptamine (5-HT), and dopamine (DA) were monitored via enzyme-linked immunosorbent assay (ELISA) to further study the sleep-improving mechanism of ZSSE. We found that sleep was effectively improved via upregulation of GABA and 5-HT and downregulation of Glu and DA. In addition, molecular mechanisms of ZSSE in improving sleep were studied by immunohistochemical analysis. The results showed that sleep was improved by regulating the expression levels of GABA receptor subunit alpha-1 (GABAARα1) and GABA acid receptor subunit gamma-2 (GABAARγ2) receptors in the hypothalamus and hippocampus tissue sections. Therefore, this work not only identified the active ingredients of ZSSE but also revealed the potential pharmacological mechanism of ZSSE for improving sleep, which may greatly stimulate the prospective development and application of ZSSE.

A bulb tubular pump unit is a horizontal tubular pump unit composed of a water pump and bulb with an electric motor installed. Electric motors, transmission equipment, and bearings are usually placed in the bulb. The bulb is located in the flow channel and has a relatively narrow space. Therefore, the shape of the bulb has a significant influence on the flow pattern and pump efficiency in the flow channel. In this study, the CFX 19.2 software was employed to optimize the bulb hydraulically according to its geometry and parameters. The research results indicate that the flow pattern at the tail of the elliptical bulb was better, the hydraulic loss at the bulb section was small, and the device efficiency was higher than that at the tail, which was round. The streamlined support had small flow resistance, minimal hydraulic loss, and a high pump unit head and efficiency. Nine schemes were selected, and the geometrical characteristics and parameters of the bulb were determined as follows: the shape of the tail of the bulb was oval, the bulb ratio was 0.96, and the shape of the support parts was streamlined. The results hold important reference significance to improve efficiency and broaden the operating conditions of bulb tubular pump devices.

Studies on the molecular mechanism of antagonistic yeasts to control apple postharvest diseases are not comprehensive enough. Our preliminary investigations screened the biocontrol effect of Hannaella sinensis, an antagonistic yeast, and discovered its control efficacy on apple blue mold decay. However, the molecular mechanism of H. sinensis-induced resistance in apple has not been studied. In this study, proteins from apple treated with H. sinensis and sterile saline were analyzed using TMT proteomics technology. It was found that H. sinensis treatment induced the expressions of apple resistance-related proteins. Among the proteins in H. sinensis-induced apple, proteins related to plant defense mechanisms, such as reactive oxygen species scavenging, improvement of plant resistance and synthesis of resistant substances, improvement of plant disease resistance, the degradation of the pathogen cell wall, cell signaling, antibacterial activity, transport of defense-related substances, and protein processing, were differentially regulated. The results of this study revealed the underlying molecular mechanisms of H. sinensis-induced apple resistance at the protein level; the results also provided a theoretical basis for the commercial application of H. sinensis.

Biological control utilizing antagonistic yeasts is an effective method for controlling postharvest diseases. Long non-coding RNAs (lncRNAs) have been found to be involved in a variety of plant growth and development processes, including those associated with plant disease resistance. In the present study, the yeast antagonist, Wickerhamomyces anomalus, was found to strongly inhibit postharvest blue mold (Penicillium expansum) and gray mold (Botrytis cinerea) decay of kiwifruit. Additionally, lncRNA high-throughput sequencing and bioinformatic analysis was used to identify lncRNAs in W. anomalus-treated wounds in kiwifruit and predict their function based on putative target genes. Our results indicate that lncRNAs may be involved in increasing ethylene (ET), jasmonic acid (JA), abscisic acid (ABA), and auxin (IAA) levels, as well as activating signal transduction pathways that regulate the expression of several transcription factors (WRKY72, WRKY53, JUB1AP2). These transcription factors (TFs) then mediate the expression of downstream, defense-related genes (ZAR1, PAD4, CCR4, NPR4) and the synthesis of secondary metabolites, thus, potentially enhancing disease resistance. Notably, by stimulating the accumulation of antifungal compounds, such as phenols and lignin, disease resistance in kiwifruit was enhanced. Our study provides new information on the mechanism underlying the induction of disease resistance in kiwifruit by W. anomalus, as well as a new disease resistance strategy that can be used to enhance the defense response of fruit to pathogenic fungi.

Patulin (PAT), mainly produced by Penicillium expansum, is a potential threat to health. In recent years, PAT removal using antagonistic yeasts has become a hot research topic. Meyerozyma guilliermondii, isolated by our group, produced antagonistic effects against the postharvest diseases of pears and could degrade PAT in vivo or in vitro. However, the molecular responses of M. guilliermondii over PAT exposure and its detoxification enzymes are not apparent. In this study, transcriptomics is used to unveil the molecular responses of M. guilliermondii on PAT exposure and the enzymes involved in PAT degradation. The functional enrichment of differentially expressed genes indicated that the molecular response mainly includes the up-regulated expression of genes related to resistance and drug-resistance, intracellular transport, growth and reproduction, transcription, DNA damage repair, antioxidant stress to avoid cell damage, and PAT detoxification genes such as short-chain dehydrogenase/reductases. This study elucidates the possible molecular responses and PAT detoxification mechanism of M. guilliermondii, which could be helpful to further accelerate the commercial application of antagonistic yeast toward mycotoxin decontamination.

Growing evidence supports that air pollution exposure has become a risk factor of type II diabetes mellitus through the induction of insulin resistance (IR), but the presented findings did not provide a consistent relationship between air pollution exposure and IR in the temporal scale and the magnitude. Reported associated with IR and air pollution exposure, branched-chain amino acids (BCAAs) in blood might modify the association between air pollution exposure and IR. We took advantage of an existing panel study on elderly people who were healthy or with pre-diabetes. Amino acids were analyzed from the serum samples using a UPLC-QQQ-MS, and the homeostasis model assessment of insulin resistance (HOMA-IR) values were calculated to represent the levels of IR in each visit. Exposures to PM2.5, NO2, SO2, CO, O3, and black carbon (BC) were estimated using data from a monitoring station. Linear mixed-effects models were applied to estimate the associations between the air pollution and HOMA-IR, as well as the modifying effects of BCAAs. We found significantly higher concentrations of BCAAs in the pre-diabetic subjects than healthy ones. The concentrations of BCAAs were all significantly associated with HOMA-IR. For subjects with high-level BCAAs, HOMA-IR was positively associated with an IQR increase in PM2.5, NO2, BC, and CO at lag day 2 and in PM2.5, SO2, NO2, BC, and CO at lag day 7. While for subjects with low-level BCAAs, there was no significant association observed at any lag day except for CO at lag day 5. The study provided evidence that circulating BCAAs may modify the relationship between air pollution exposure and the level of insulin resistance in humans.

Blue mold, caused by Penicillium expansum, is the most destructive fungal disease of apples and causes great losses during the post-harvest storage of the fruit. Although some apple cultivars are resistant to P. expansum, there has been little information on the molecular mechanism of resistance. In this study, differential proteomic analysis was performed on apple samples infected and uninfected with P. expansum. Parallel reaction monitoring (PRM) technology was used to target and verify the expression of candidate proteins. The label-free technique identified 343 differentially expressed proteins, which were mainly associated with defense responses, metal ion binding, stress responses, and oxidative phosphorylation. The differential expression of enzymes related to reactive oxygen species (ROS) synthesis and scavenging, the activation of defense-related metabolic pathways, and the further production of pathogenesis-related proteins (PR proteins) during P. expansum infection in apples, and direct resistance to pathogen invasion were determined. This study reveals the mechanisms of apple response at the proteomic level with 9 h of P. expansum infection.

The composition of microbial communities can directly affect fruit quality, health status, and storability. The present study characterized the epiphytes and endophytes of “Hongyang” and “Cuiyu” kiwifruit at harvest under grown under open-field (OF) and rain-shelter (RS) cultivation systems. Disease incidence in kiwifruit was significantly lower (p < 0.05) under the RS system than it was under the OF system. High-throughput sequencing [16S V3-V4 ribosomal region and the fungal internal transcribed spacer (ITS2)] was conducted to compare the composition of the epiphytic and endophytic microbial community of kiwifruit under the two cultivation systems. Results indicated that the abundance of Actinobacteria, Bacteroidetes, Enterobacteriales, Acetobacterales, Sphingomonas, Pseudomonas, and Sphingobacterium was higher under the RS system, relative to the OF system, while the abundance of Capnodiales, Hypocreales, Vishniacozyma, and Plectosphaerella was also higher under the RS system. Some of these bacterial and fungal taxa have been reported to as act as biocontrol agents and reduce disease incidence. Notably, the α-diversity of the epiphytic bacterial and fungal communities on kiwifruit was higher under RS cultivation. In summary, RS cultivation reduced natural disease incidence in kiwifruit, which may be partially attributed to differences in the structure and composition of the microbial community present in and on kiwifruit.

Transcription factors play a key role in Penicillium expansum infection process. Although the crucial characteristics of some transcription factors of pathogenic fungi have been found, many transcription factors involved in P. expansum infections have not been explored and studied. This study aimed to screen the transcription factors of P. expansum involved in postharvest pear infections by ATAC-seq analysis and to analyze the differentially expressed peak-related genes by GO enrichment and KEGG pathway analysis. Our results found the up-regulation of differentially expressed peak-related genes involved in the MAPK signaling pathway, pentose phosphate pathway, starch and sucrose metabolism, and pentose and glucuronate interconversions. Our study especially confirmed the differential regulation of transcription factors MCM1, Ste12 and gene WSC in the MAPK signaling pathway and PG1, RPE1 in the pentose and glucuronate interconversions pathway. These transcription factors and related genes might play an essential role in pear fruit infection by P. expansum. RT-qPCR validation of twelve expressed peak-related genes in P. expansum showed that the expression levels of these twelve genes were compatible with the ATAC-Seq. Our findings might shed some light on the regulatory molecular networks consisting of transcription factors that engaged in P. expansum invasion and infection of pear fruits.

To study the mechanism by which Pichia anomala induced with chitosan (1% w/v) controls blue mold disease in table grapes caused by Penicillium expansum, this study evaluated alterations in three yeast enzymatic activities. The changes in the five primary disease defense-related enzymes and two non-enzyme activities of table grapes were assayed. The results of the study showed that chitosan (1% w/v) significantly increased the yeast β-1,3-glucanase, catalase (CAT), and malondialdehyde (MDA) activities. Furthermore, P. anomala alone or induced with chitosan (1% w/v) significantly increased the table grapes enzymatic activities of Polyphenol oxidase (PPO), phenylalanine (PAL), peroxidase (POD), and catalase (CAT) compared to the control. The RT-qPCR results also confirmed that the genes of these major disease defense enzymes were up-regulated when the table grapes were treated with P. anomala. The highest results were recorded when the fruit was treated by yeast induced with chitosan (1% w/v). The phenolic compounds, in addition to their nutritional value, can also increase the antimicrobial properties of table grapes. The current experiment determined that the total phenol and flavonoid contents of table grapes showed the highest results for fruits treated by P. anomala induced with chitosan compared with the control. Generally, the increment of these fruit enzymatic and non-enzymatic activities shows improved table grape defense against the pathogenic fungus. The induction of the yeast with chitosan also increases its bio-control efficacy against the pathogen. This study will enable future detailed investigation in the yeast pathogen control mechanisms and the use of yeasts as bio-pesticides.

Zearalenone (ZEN) is produced by Fusarium spp. and is widely found in moldy wheat, corn, and other grains. ZEN has a strong toxicity and causes reproductive and immune disorders and estrogenic syndrome in animals and humans. Biodegradation has been demonstrated as an efficient way to control the hazardous effect of ZEN. A promising way to apply biodegradation in feed is to introduce anaerobic ZEN-degrading microorganisms, which can function during the digestion process in animal intestines. The aim of this study was to isolate anaerobic ZEN-degrading bacteria from anaerobic environments. A strain named F39 was isolated from animal intestinal contents and had a ZEN-degradation rate of 87.35% in 48 h to form trace amount of α- and β-zearalenol. Based on the morphological and physiological properties and phylogenetic analysis of 16S rRNA and rpoB gene sequences, F39 was identified as Clostridium sporogenes. The optimum temperature for the growth of F39 was 37 °C, the optimum pH was 7.0, and the most suitable carbon source was beef extract, while the optimal conditions for the degradation of ZEN were as follows: 35 °C, pH 7.0, and GAM medium. ZEN was degraded by F39 with a high efficiency in the concentration range of 1–15 mg/L. The bioactive factors responsible for ZEN degradation were mainly distributed intracellularly. F39 can degrade most of the ZEN present, but a small amount is broken down into two secondary metabolites, α- and β-zearalenol, and the toxicity of the degradation products is reduced. With an efficiency of 49%, F39 can more effectively degrade ZEN in wheat-based feedstuffs than in other feedstuff, and the degradation efficiency was pH related. To the best of our knowledge, this is the first report of Clostridium sporogenes F39’s ability to maintain the biodegradation potentials.

Gray mold decay is a widespread postharvest disease in tomato that results from infection by the pathogen Botrytis cinerea, leading to huge economic losses. The objective of this study was to select the most effective antagonistic yeast to control tomato gray mold from six potential biocontrol agents and to investigate the possible control mechanism. The results showed that the yeast Wickerhamomyces anomalus was the most effective in inhibiting B. cinerea among the six strains both in vivo and in vitro on tomato, with a colony diameter of 11 mm, a decay diameter of 20 mm, and the lowest decay incidence (53%)—values significantly smaller and lower than the values recorded for the control group and the other yeasts. The efficacy of the control depended on the increase in yeast concentration, and the decay incidence and lesion diameter were reduced to 31%, 28% and 7 mm, 6 mm, respectively, when treated with 1 × 108 and 1 × 109 cells/mL W. anomalus. In addition, W. anomalus was able to rapidly colonize and stably multiply in tomato, occupying the space to control pathogen infection. W. anomalus was also able to motivate the defense mechanism of tomato with stimulation of defense-related enzymes PPO, POD, APX, and SOD and promotion of the content of total phenols and flavonoid compounds. All these results suggest that W. anomalus exhibited exceptional ability to control gray mold in tomato.

A better understanding of the mode of action of postharvest biocontrol agents on fruit surfaces is critical for the advancement of successful implementation of postharvest biocontrol products. This is due to the increasing importance of biological control of postharvest diseases over chemical and other control methods. However, most of the mechanisms involved in biological control remain unknown and need to be explored. Yarrowia lipolytica significantly inhibited blue mold decay of apples caused by Penicillium expansum. The findings also demonstrated that Y. lipolytica stimulated the activities of polyphenoloxidase, peroxidase, chitinase, l-phenylalanine ammonia lyase involved in enhancing defense responses in apple fruit tissue. Proteomic and transcriptomic analysis revealed a total of 35 proteins identified as up- and down-regulated in response to the Y. lipolytica inducement. These proteins were related to defense, biotic stimulus, and stress responses, such as pathogenesis-related proteins and dehydrin. The analysis of the transcriptome results proved that the induced resistance was mediated by a crosstalk between salicylic acid (SA) and ethylene/jasmonate (ET/JA) pathways. Y. lipolytica treatment activated the expression of isochorismate synthase gene in the SA pathway, which up-regulates the expression of PR4 in apple. The expression of 1-aminocyclopropane-1-carboxylate oxidase gene and ET-responsive transcription factors 2 and 4, which are involved in the ET pathway, were also activated. In addition, cytochrome oxidase I, which plays an important role in JA signaling for resistance acquisition, was also activated. However, not all of the genes had a positive effect on the SA and ET/JA signal pathways. As transcriptional repressors in JA signaling, TIFY3B and TIFY11B were triggered by the yeast, but the gene expression levels were relatively low. Taken together, Y. lipolytica induced the SA and ET/JA signal mediating the defense pathways by stimulating defense response genes, such as peroxidase, thaumatin-like protein, and chitinase 4-like, which are involved in defense response in apple.[Graphic: see text] Copyright © 2017 The Author(s) This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Antagonistic yeasts can inhibit fungal growth. In our previous research, Meyerozyma guilliermondii, one of the antagonistic yeasts, exhibited antagonistic activity against Penicillium expansum. However, the mechanisms, especially the molecular mechanisms of inhibiting activity of M. guilliermondii, are not clear. In this study, the protein expression profile and transcriptome characterization of P. expansum induced by M. guilliermondii were investigated. In P. expansum induced by M. guilliermondii, 66 proteins were identified as differentially expressed, among them six proteins were upregulated and 60 proteins were downregulated, which were associated with oxidative phosphorylation, ATP synthesis, basal metabolism, and response regulation. Simultaneously, a transcriptomic approach based on RNA-Seq was applied to annotate the genome of P. expansum and then studied the changes of gene expression in P. expansum treated with M. guilliermondii. The results showed that differentially expressed genes such as HEAT, Phosphoesterase, Polyketide synthase, ATPase, and Ras-association were significantly downregulated, in contrast to Cytochromes P450, Phosphatidate cytidylyltransferase, and Glutathione S-transferase, which were significantly upregulated. Interestingly, the downregulated differentially expressed proteins and genes have a corresponding relationship; these results revealed that these proteins and genes were important in the growth of P. expansum treated with M. guilliermondii.

Transcriptome analysis (TA) was conducted to characterize the transcriptome changes in postharvest disease-related genes of table grapes following treatment with Pichia anomala induced with chitosan (1% w/v). In the current study, the difference in the gene expression of table grapes after treatment with P. anomala induced with chitosan and that of a control group was compared 72 h post-inoculation. The study revealed that postharvest treatment of table grapes with P. anomala induced with chitosan could up-regulate genes that have a pivotal role in the fruit’s disease defense. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) results also confirmed that GO terms and the KEGG pathways, which have pivotal roles in plant disease resistance, were significantly enriched. The up-regulated genes of the treatment group have a unique function in the fruit’s disease resistance compared to the control group. Generally, most genes in the plant–pathogen interaction pathway; the plant Mitogen-activated protein kinase (MAPK) signaling pathway; the plant hormone signal transduction pathway; the pathway of glutathione metabolism; the pathway of phenylalanine, tyrosine, and tryptophan biosynthesis; and the pathway of flavonoid biosynthesis were all up-regulated. These up-regulations help the fruit to synthesize disease-resistant substances, regulate the reactive oxygen species (ROS), enhance the fruit cell wall, and enrich hormone signal transduction during the pathogen’s attack. This study is useful to overcome the lags in applying transcriptomics technology in postharvest pathology, and will provide insight towards developing other alternative methods to using bio-pesticides to control postharvest diseases of perishables.

Patulin (PAT) is a mycotoxin produced by some Penicillium, Aspergillus, and Byssochlamys species. Rhodotorula mucilaginosa is able to degrade PAT in vivo as well as in vitro, up till date, the process and molecular mechanism(s) involved patulin degradation still remains unknown. Protein lysine crotonylation (Kcr) plays an important role in regulating chromatin dynamics, gene expression, and metabolic pathways in mammals and eukaryotes. Investigation of the Kcr changes accompanying degradation of patulin in R. mucilaginosa were observed to investigate the mechanisms of patulin inhibition. Tandem mass tag (TMT) labeling and Kcro affinity enrichment, followed by high-resolution LC-MS/MS analysis, were used to perform quantitative lysine crotonylome analysis on R. mucilaginosa. Consequently, 1691 lysine crotonylation sites in 629 protein groups were identified, among which we quantified 1457 sites in 562 proteins. Among the quantified proteins, 79 and 46 crotonylated proteins were up-regulated and down-regulated, respectively. The differentially up expressed modified proteins were mainly involved in tricarboxylic acid cycle and gluconeogenic pathway. The differentially down expressed Kcr proteins were mainly classified to ribosome and carbohydrate transport and metabolism. Bioinformatic analyses were performed to annotate the quantifiable lysine crotonylated targets. Moreover, interaction networks and high confidence domain architectures of crotonylated proteins were investigated with the aid of bioinformatic tools, and these results showed that there was an increase in the number of yeasts with crotonylated proteins. The results also provided information on the various roles of crotonylation, which are involved in PAT degradation.

The efficacy of Sporidiobolus pararoseus Y16 in controlling postharvest blue mold caused by Penicillium expansum on apples and the defense response involved were evaluated. The results suggested that the decay incidence of blue mold of apples treated by S. pararoseus Y16 was significantly reduced compared with the control. In vitro testing indicated that germination of spores and germ tube length of P. expansum were markedly inhibited by S. pararoseus Y16. Meanwhile, polyphenol oxidase (PPO), peroxidase (POD), phenylalanine ammonia lyase (PAL), and catalase (CAT) activities and several pathogenesis-related (PR) gene expression levels (including PR3, PR4, PR5, and PR9) were determined. In apples, the activities of PPO, POD, CAT, and PAL were significantly induced by S. pararoseus Y16 treatment compared with the control fruits. The relative expression levels of PR3 and PR4 were significantly induced at 4 and 6 d, while PR5 was significantly induced at 4 and 6 d and PR9 was significantly induced at 4 d. Therefore, the reduction in apple fruit decay by S. pararoseus Y16 treatment could be related to the increased activities of related enzymes and proteins involved in the defense against pathogens, which suggest that S. pararoseus Y16 is a potential antagonistic yeast.

Synthetic fungicides are commonly employed for the control of postharvest diseases of fruits. However, due to health concerns about the use of these chemicals, alternative control methods including biocontrol based on antagonistic yeasts are gaining in popularity. In this study, we investigated the effects of two biocontrol yeasts, Rhodotorula mucilaginosa strain 3617 and Rhodotorula kratochvilovae strain LS11, on blue mold and patulin (PAT) contamination caused by Penicillium expansum strains PY and FS7 in artificially inoculated Fuji apples stored at 20°C for 9 days. To correlate the development of the P. expansum strains in yeast-treated and untreated apples with PAT production, we quantified their biomass in the infected fruits using a recently published quantitative real-time polymerase chain reaction method based on specific primers for patF, a gene from P. expansum that is involved in PAT biosynthesis. Both yeasts significantly reduced the disease incidence caused by the two strains of P. expansum up to 5–7 days of incubation, and lowered their biomass and the progression of symptoms up to 9 days. Interestingly, both yeasts strains increased the rate of PAT production (expressed as ng patulin/μg fungal DNA) by the two pathogenic strains. Nevertheless, both biocontrol agents reduced the total PAT contamination, especially in the case of P. expansum strain FS7, the higher PAT producer of the two tested P. expansum strains. Comparing between the yeast strains, R. kratochvilovae LS11 was more effective than R. mucilaginosa 3617 for the control of P. expansum.

Green mold disease, a common citrus post-harvest disease caused by Penicillium digitatum, has an unresolved initial infection mechanism. Understanding the infection mechanism leads to the development of potential controls and preventive measures against the disease. The present study aimed to delineate the infection mechanism by investigating spore germination, changes of organic molecules and enzyme activity, and differential expression of genes in the P. digitatum infection. P. digitatum spore germination was observed by a pathology section scanner and it was found that in vivo germination was 3 h behind the in vitro germination. In addition, cell wall degrading enzymes and soluble sugar and titratable acid content during the infection process measured dynamically. The level of pectinase reached its maximum of 6067 U/g before 48 hpi, while cellulase increased rapidly after 48 hpi. The soluble sugar and organic acid content increased considerably with the progression of the infection. The transcriptomic profile of P. digitatum before and after infection was analyzed by RNA-seq. The genes related to cell wall degrading enzymes were significantly up-regulated and annotated to participate in two major carbon source synthesis pathways. The study delineated the initial infection mechanism of P. digitatum which eventually opened the gate way for the development of new control strategies in the future.

Blue mold, caused by Penicillium expansum, is an important postharvest disease of apple, and can result in significant economic losses. The present study investigated the interaction between P. expansum and wounded apple fruit tissues during the early stages of the infection. Spores of P. expansum became activated one hour post-inoculation (hpi), exhibited swelling at 3 hpi, and the germ tubes were found entering into apple tissues at 6 hpi. RNA-seq was performed on samples of P. expansum and apple fruit tissue collected at 1, 3, and 6 hpi. The main differentially expressed genes (DEGs) that were identified in P. expansum were related to interaction, cell wall degradation enzymes, anti-oxidative stress, pH regulation, and effectors. Apple tissues responded to the presence of P. expansum by activating pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) at 1 hpi, then activated effector-triggered immunity (ETI) at 3 hpi. This research provides new information on the interaction between P. expansum and apple fruit tissue at an early stage of the infection process.

20-Hydroxy-3-oxolupan-28-oic acid (HOA), a lupane-type triterpene, was obtained from the leaves of Mahonia bealei, which is described in the Chinese Pharmacopeia as a remedy for inflammation and related diseases. The anti-inflammatory mechanisms of HOA, however, have not yet been fully elucidated. Therefore, the objective of this study was to characterize the molecular mechanisms of HOA in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. HOA suppressed the release of nitric oxide (NO), pro-inflammatory cytokine tumor necrosis factor α (TNF-α), and interleukin 6 (IL-6) in LPS-stimulated RAW264.7 macrophages without affecting cell viability. Quantitative real-time reverse-transcription polymerase chain reaction (RT-qPCR) analysis indicated that HOA also suppressed the gene expression of inducible NO synthase (iNOS), TNF-α, and IL-6. Further analyses demonstrated that HOA inhibited the phosphorylation of upstream signaling molecules, including p85, PDK1, Akt, IκBα, ERK, and JNK, as well as the nuclear translocation of nuclear factor κB (NF-κB) p65. Interestingly, HOA had no effect on the LPS-induced nuclear translocation of activator protein 1 (AP-1). Taken together, these results suggest that HOA inhibits the production of cytokine by downregulating iNOS, TNF-α, and IL-6 gene expression via the downregulation of phosphatidylinositol 3-kinase (PI3K)/Akt and mitogen-activated protein kinases (MAPKs), and the inhibition of NF-κB activation. Our findings indicate that HOA could potentially be used as an anti-inflammatory agent for medical use.

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Ochratoxin A (OTA) contamination has been established as a world-wide problem. In this study, the strains with the ability of OTA production were screened by analyzing the green fluorescence of the isolates colonies from the grapes in Zhenjiang with 365 nm UV light and confirmed by HPLC with fluorescent detection (HPLC-FLD). The results showed that seven isolates acquired the characteristic of the fluorescence, of which only five showed the ability of OTA production as confirmed by HPLC-FLD analysis. The five OTA-producing strains were identified based on comparative sequence analysis of three conserved genes (ITS, BenA and RPB2) of the strains, and they are Talaromyces rugulosus (O1 and Q3), Penicillium commune (V5-1), Penicillium rubens (MQ-5) and Aspergillus aculeatus (MB1-1). There are two Penicillium species of the five OTA-producing strains and our study is the first to report that P. rubens, T. rugulosus and A. aculeatus can produce OTA. This work would contribute to comprehensively understanding the fungi with an OTA-producing ability in grapes before harvest and then take effective measures to prevent OTA production.

In this work, we examined the mechanisms involved in the degradation of patulin by Pichia caribbica. Our results indicate that cell-free filtrate of P. caribbica reduced patutlin content. The heat-killed cells could not degrade patulin. However, the live cells significantly reduced the concentration of the patulin. In furtherance to this, it was observed that patulin was not detected in the broken yeast cells and cell wall. The addition of cycloheximide to the P. caribbica cells decreased the capacity of degradation of patulin. Proteomics analyses revealed that patulin treatment resulted in an upregulated protein which was involved in metabolism and stress response processes. Our results suggested that the mechanism of degradation of patulin by P. caribbica was not absorption; the presence of patulin can induce P. caribbica to produce associated intracellular and extracellular enzymes, both of which have the ability to degrade patulin. The result provides a new possible method that used the enzymes produced by yeast to detoxify patulin in food and feed.