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In order to obtain endophytic selenium-rich yeast from Lycium barbarum with strong ability producing alcohol and aroma, and then promote the selenium-rich fermentation products and improve the flavor, this study took the endophytic yeast of L. barbarum stored in the laboratory as experimental materials, and the yeast with strong selenium-rich ability was obtained through the initial screening of selenium-resistant method and red selenium methods. Atmospheric room temperature plasma (ARTP) technology was used to obtain mutant strains and through TTC medium, Duchenne tubule fermentation, alcohol production, ester and acid production, tolerance and other experiments to screen out the target strains of aroma enhancement and high alcohol production. The content of selenium in yeast cell and fermentation liquid was determined by hydride generation atomic fluorescence spectrometry. The results showed that M1 Saccharomyces cerevisiae and M16 Clavispora lusitaniae were obtained by primary selenium screening, the mutagenic strain M1-5 of S. cerevisiae with higher alcohol production capacity and the mutagenic strain M16-28 of C. lusitaniae with higher aroma production capacity were obtained through ARTP mutagenesis. The alcohol production capacity of M1-5 increased by 38.07% compared with M1, and the ester production capacity of M16-28 increased by 107.62% compared with M16, and the genetic stability was good. The selenium contents in the intracellular and fermentation broth of the M1-5 strains were 204.8 and 1158 μg/L, respectively, and in M16-28 were 666.45μg/L and 1830 μg/L, respectively. Therefore, the selenium enrichment capacity of M16-28 was higher than that of M1-5, and the selenium content of fermentation liquid was extremely significantly higher than that of yeast cells (P

Abstract Background The macrolide antibiotic avermectin, a natural product derived from Streptomyces avermitilis, finds extensive applications in agriculture, animal husbandry and medicine. The mtrA (sav_5063) gene functions as a transcriptional regulator belonging to the OmpR family. As a pleiotropic regulator, mtrA not only influences the growth, development, and morphological differentiation of strains but also modulates genes associated with primary metabolism. However, the regulatory role of MtrA in avermectin biosynthesis remains to be elucidated. Results In this study, we demonstrated that MtrA, a novel OmpR-family transcriptional regulator in S. avermitilis, exerts global regulator effects by negatively regulating avermectin biosynthesis and cell growth while positively controlling morphological differentiation. The deletion of the mtrA gene resulted in an increase in avermectin production, accompanied by a reduction in biomass and a delay in the formation of aerial hyphae and spores. The Electrophoretic Mobility Shift Assay (EMSA) revealed that MtrA exhibited binding affinity towards the upstream region of aveR, the intergenic region between aveA1 and aveA2 genes, as well as the upstream region of aveBVIII in vitro. These findings suggest that MtrA exerts a negative regulatory effect on avermectin biosynthesis by modulating the expression of avermectin biosynthesis cluster genes. Transcriptome sequencing and fluorescence quantitative PCR analysis showed that mtrA deletion increased the transcript levels of the cluster genes aveR, aveA1, aveA2, aveC, aveE, aveA4 and orf-1, which explains the observed increase in avermectin production in the knockout strain. Furthermore, our findings demonstrate that MtrA positively regulates the cell division and differentiation genes bldM and ssgC, while exerting a negative regulatory effect on bldD, thereby modulating the primary metabolic processes associated with cell division, differentiation and growth in S. avermitilis, consequently impacting avermectin biosynthesis. Conclusions In this study, we investigated the negative regulatory effect of the global regulator MtrA on avermectin biosynthesis and its effects on morphological differentiation and cell growth, and elucidated its transcriptional regulatory mechanism. Our findings indicate that MtrA plays crucial roles not only in the biosynthesis of avermectin but also in coordinating intricate physiological processes in S. avermitilis. These findings provide insights into the synthesis of avermectin and shed light on the primary and secondary metabolism of S. avermitilis mediated by OmpR-family regulators.

Abstract Background Vitamin B12 (VB12) has a wide range of applications and high economic value. In this study, a new strain with high VB12 production potential, Ensifer adhaerens S305, was identified in sewage. Because E. adhaerens strains have become the main strains for VB12 production via fermentation in recent years, the directional modification of the S305 strain to obtain a strain suitable for the industrial production of VB12 has great potential and commercial value. Results 16S rRNA and genome-wide phylogenetic tree analysis combined with average nucleotide identity (ANI) analysis showed that the high-yielding VB12 strain was a E. adhaerens strain and that its VB12 synthesis pathway genes were highly similar to related genes of strains of this and other species, including E. adhaerens Casida A, Pseudomonas denitrificans SC 510, and E. adhaerens Corn53. High-pressure liquid chromatography (HPLC) results indicated that the VB12 yields of the S305 strain were more than double those of the Casida A strain under different medium components. Multiple genes with significantly upregulated and downregulated transcription were identified by comparing the transcription intensity of different genes through transcriptome sequencing. KEGG enrichment analysis of the porphyrin metabolism pathway identified 9 significantly upregulated and downregulated differentially expressed genes (DEGs) in the VB12 synthesis pathway, including 7 transcriptionally upregulated genes (cobA, cobT, hemA, cobJ, cobN, cobR, and cobP) that were episomally overexpressed in the Casida A strain. The results showed that the VB12 yield of the overexpressed strain was higher than that of the wild-type strain. Notably, the strains overexpressing the cobA and cobT genes exhibited the most significant increases in VB12 yield, i.e., 31.4% and 24.8%, respectively. The VB12 yield of the S305 strain in shake-flask culture was improved from 176.6 ± 8.21 mg/L to 245.6 ± 4.36 mg/L by integrating the cobA and cobT genes into the strain. Conclusion Phylogenetic tree and ANI analysis showed that the Ensifer and Sinorhizobium strains were quite different at the genome level; the overexpression and integrated expression of significantly upregulated genes in the VB12 synthesis pathway could increase the yield of VB12, further improving the VB12 yield of the E. adhaerens S305 strain.

As one of the important lakes in the “One Lake and Two Seas” of the Inner Mongolia Autonomous Region, the monitoring of water quality in Lake Daihai has attracted increasing attention, and the concentration of chlorophyll-a directly affects the water quality, making the monitoring of chlorophyll-a concentration in Lake Daihai particularly crucial. Traditional methods of monitoring chlorophyll-a concentration are not only inefficient but also require significant human and material resources. Remote sensing technology has the advantages of wide coverage and short update cycles. For lakes such as Daihai with a high salinity content, salinity is considered a key factor when inverting the concentration of chlorophyll-a. In this study, machine learning models, including model stacking from ensemble learning, a ridge regression model, and a random forest model, were constructed. After comparing the training accuracy of the three models on Zhuhai-1 satellite data, the random forest model, which had the highest accuracy, was selected as the final training model. By comparing the accuracy changes before and after adding salinity factors to the random forest model, a high-precision model for inverting chlorophyll-a concentration in hypersaline lakes was obtained. The research results show that, without considering the salinity factor, the root mean square error (RMSE) of the model was 0.056, and the coefficient of determination (R2) was 0.64, indicating moderate model performance. After adding the salinity factor, the model accuracy significantly improved: the RMSE decreased to 0.047, and the R2 increased to 0.92. This study provides a solid basis for the application of remote sensing technology in hypersaline aquatic environments, confirming the importance of considering salinity when estimating chlorophyll-a concentration in hypersaline waters. This research helps us gain a deeper understanding of the water quality and ecosystem evolution in Daihai Lake.

Stevioside is a secondary metabolite of diterpenoid glycoside production in plants. It has been used as a natural sweetener in various foods because of its high sweetness and low-calorie content. In this study, we constructed a Saccharomyces cerevisiae strain for the complete synthesis of stevioside using a metabolic engineering strategy. Firstly, the synthesis pathway of steviol was modularly constructed in S. cerevisiae BY4742, and the precursor pathway was strengthened. The yield of steviol was used as an indicator to investigate the expression effect of different sources of diterpene synthases under different combinations, and the strains with further improved steviol yield were screened. Secondly, glycosyltransferases were heterologously expressed in this strain to produce stevioside, the sequence of glycosyltransferase expression was optimized, and the uridine diphosphate-glucose (UDP-Glc) supply was enhanced. Finally, the results showed that the strain SST-302III-ST2 produced 164.89 mg/L of stevioside in a shake flask experiment, and the yield of stevioside reached 1104.49 mg/L in an experiment employing a 10 L bioreactor with batch feeding, which was the highest yield reported. We constructed strains with a high production of stevioside, thus laying the foundation for the production of other classes of steviol glycosides and holding good prospects for application and promotion.

In order to improve the accuracy of vehicle trajectories and ensure driving safety, and considering the differences in driver behavior and the impact of these differences on vehicle trajectories, a vehicle trajectory-prediction method (DBC-Informer) based on the categorization of driver behavior is proposed: firstly, the characteristic driver feature data are extracted through data preprocessing; secondly, descriptive statistical data are obtained through the classification of the driver’s behavior into categories; finally, based on the Informer model, a two-layer driver category trajectory-prediction network architecture is established, which inputs the vehicle trajectories of different driving types into independent prediction sub-networks, respectively, to realize the accurate prediction of vehicle trajectories. The experimental results show that the MAE and MSE values of trajectory prediction of the DBC-Informer model in different time domains are much smaller than those of other comparative models, and the improvement of accuracy is more obvious in the long-term domain trajectory-prediction task scenario, and the increase in prediction error of the DBC-Informer model is significantly reduced after the prediction time exceeds 1 s. The on-line behavioral categorization is achieved by comparing different categorization models; it reaches 98% in classification accuracy and, according to the results of ablation experiments, the addition of the driver behavior-classification method to the prediction model improves the accuracy of prediction in longitudinal and lateral motion by 56% and 61%, respectively, which verifies the effectiveness of the driver behavior-classification method. It can be seen that the DBC-Informer model can more accurately portray the effects of different driving behaviors on vehicle trajectories and improve the accuracy of vehicle trajectory prediction, which provides important data support for vehicle warning systems.

Both heavy metals and antibiotics exert selection pressure on bacterial resistance, and as they are commonly co-contaminated in the environment, they may play a larger role in bacterial resistance. This study examined how breeding cycles affect antibiotic resistance genes (ARGs) in chicken manure and the surrounding topsoils at 20, 50, 100, 200, and 300 m from twelve typical laying hen farms in the Ningxia Hui Autonomous Region of northwest China. Six antibiotics, seven heavy metals, ten mobile genetic elements (MGEs), and microbial community affected the ARGs profile in chicken dung and soil samples. Tetracycline antibiotic residues were prevalent in chicken manure, as were relatively high content of aureomycin during each culture period. Zinc (Zn) content was highest among the seven heavy metals in chicken feces. Chicken dung also enriched aminoglycosides, MLSB, and tetracycline ARGs, notably during brooding and high production. The farm had a minimal influence on antibiotics in the surrounding soil, but its effect on ARGs and MGEs closer to the farm (50 m) was stronger, and several ARGs and MGEs increased with distance. Manure microbial composition differed dramatically throughout breeding cycles and sampling distances. ARGs were more strongly related with antibiotics and heavy metals in manure than soil, whereas MGEs were the reverse. Antibiotics, heavy metals, MGEs, and bacteria in manure accounted 12.28%, 22.25%, 0.74%, and 0.19% of ARGs composition variance, respectively, according to RDA and VPA. Bacteria (2.89%) and MGEs (2.82%) only affected soil ARGs composition. These findings showed that heavy metals and antibiotics are the main factors affecting faecal ARGs and bacteria and MGEs soil ARGs. This paper includes antibiotic resistance data for large-scale laying hen husbandry in northwest China and a theoretical framework for decreasing antibiotic resistance.

Cinnamomum burmannii is a cinnamomum plant rich in natural D-borneol. Natural D-borneol is a bicycle monoterpenoid compound widely used in the food, pharmaceutical, and cosmetic industries. Therefore, analyzing the biosynthesis mechanism of natural D-borneol in C. burmannii at the molecular level is helpful for directional breeding in the future and further development and utilization of C. burmannii and its related gene resources. In our study, 76 genes related to terpene metabolism were analyzed through third-generation sequencing and second-generation sequencing. Of these genes, 57 were associated with the synthesis of the terpenoid skeleton, and 19 belonged to terpenoid synthase, including four monoterpenoid synthases, seven sesquiterpenoid synthases, and eight diterpenoid synthases. Two genes in diterpenoid synthase were differentially expressed in high D-borneol and low D-borneol plants. It was speculated that these two genes might be related to D-borneol synthesis. How these two genes participate in the synthesis of D-borneol needs further study.

Abstract It is challenging to match the mutual interactions between implant and host because the biomaterials usually cannot actively adjust their performance to the changing microenvironment. Surface potential is one of the critical factors affecting the bioactivity of biomaterials, but it is difficult to be directly controlled in vivo. Magnetic stimulation has attracted much attention due to its deep penetrability, good reliability, and convenient operability. Here, titanium dioxide (TiO2) nanotubes and Terfenol‐D/P(VDF‐TrFE) composite film are prepared by anodic oxidation and solution casting methods on opposite sides of a titanium sheet, respectively. Terfenol‐D magnetostrictive microparticles deform under a magnetic field, generating surface potential on the P(VDF‐TrFE) piezoelectric matrix through magneto‐electric coupling. Correspondingly, equal opposite charges are induced on the surface of TiO2 nanotubes. Stem cells cultured on TiO2 nanotubes show that cell adhesion, proliferation, and differentiation abilities can be regulated by magnetic strength, which correlates with the absorption of charged proteins. Therefore, a cascade coupling of magnetic, mechanical, electric, biochemical, and cellular effects is established. This work demonstrates the feasibility of regulating the bioactivity of biomaterials in vivo through a magnetic field.

Liquidambar formosana is a multipurpose tree species native to China. There has been increasing interest in L. formosana due to its leaves being rich in shikimic acid, which plays a key role in the synthesis of the antiviral drug oseltamivir phosphate. Here, shikimic acid content (SAC) and other breeding traits, including tree height (HT), diameter at breast height (DBH), height to crown base (HCB), individual tree volume (VOL), leaf color (LC) and stem straightness degree (SSD), for 387 families of 19 provenances were evaluated in a provenance–family trial of L. formosana to estimate genetic parameters and reveal geographical variation patterns and, ultimately, screen out superior provenances and families. Differences among provenances and families were significant for all tested traits, indicating a high potential for selective breeding. Broad-sense heritabilities of provenance (hp2 = 0.19–0.57) and family (hf2 = 0.16–0.31) were moderate for most traits. Moderate to strong genetic correlations were found among HT, DBH, VOL, HCB and LC (rA = 0.339–0.982), while adverse correlations (rA = −0.494 to −0.816) were observed between SAC and growth traits. All target traits, excluding SSD, exhibited clinal variation in response to latitudinal gradients, and a clustering heatmap divided the 19 provenances into three groups. For single-trait selection, SAC persistently had the highest genetic gains (85.14%–163.57%). A weighted index based on breeding values was used to concomitantly improve SAC, HT and DBH. At a selection rate of 25%, the genetic gains at the provenance and family levels for SAC were 36.42% and 73.52%, and those for core growth traits ranged from −2.29% to 3.49% and 4.05% to 4.47%, respectively. As far as we know, this is the first study in L. formosana to explore the inheritance of SAC and its correlations with other traditional breeding traits. The genetic parameter estimations contribute to a better understanding of the genetic basis of SAC, and the superior provenances and families obtained lay a material foundation for the development of new varieties rich in shikimic acid, thereby promoting the in-depth exploitation and utilization of germplasm resources of L. formosana.

Glucose is an important carbon source for microbial growth, and its content in infertile soils is essential for the growth of bacteria. Since the mechanism of oligotrophic bacterium adaptation in barren soils is unclear, this research employed RNA-seq technology to examine the impact of glucose concentration on the oligotrophic bacterium B. subtilis BS-G1 in soil affected by desertification. A global transcriptome analysis (RNA-Seq) revealed that the significantly differentially expressed genes (DEGs) histidine metabolism, glutamate synthesis, the HIF-1 signaling pathway, sporulation, and the TCA cycle pathway of B. subtilis BS-G1 were significantly enriched with a 0.015 g/L glucose concentration (L group), compared to a 10 g/L glucose concentration (H group). The DEGs amino acid system, two-component system, metal ion transport, and nitrogen metabolism system of B. subtilis BS-G1 were significantly enriched in the 5 g/L glucose concentration (M group), compared with the H group. In addition, the present study identified the regulation pattern and key genes under a low-glucose environment (7 mRNAs and 16 sRNAs). This study primarily investigates the variances in the regulatory pathways of the oligotrophic B. subtilis BS-G1, which holds substantial importance in comprehending the mechanism underlying the limited sugar tolerance of oligotrophic bacteria.

Streptomyces fradiae sf106 is a type of actinomycete that can produce abundant secondary metabolites, making it a natural cell factory for drug synthesis. In order to comprehensively understand the genomic profile of Streptomyces fradiae sf106 and its potential for producing secondary metabolites, a combination of several methods was used to perform whole-genome sequencing of sf106. The results showed that sf106 is most closely related to Streptomyces xinghaiensis S187; the average nucleotide identity and average amino acid identity of sf106 and S187 were more than 96%. The genome size of sf106 is approximately 7300 kb, the GC content is greater than 72%, and more than 6700 coding sequences (CDS) were identified. Analysis of mobile genetic elements revealed the presence of a large number of horizontally transferred genes in Streptomyces fradiae sf106, which contribute to microbial diversity. Through antiSMASH prediction, 22 secondary metabolite gene clusters were obtained, which had great potential to generate polyketide metabolites. By examining the data, it was found that the genes contained in cluster 9 were similar to those involved in tylosin synthesis. Non-targeted metabolome sequencing revealed that a total of 1855 identifiable metabolites were produced in the fermentation broth, and the majority of metabolites showed highly significant differences in mean relative abundance between the groups. The identified metabolites were compared against the KEGG compound database to obtain metabolite classifications, mainly including Biological Roles, Phytochemical Compounds, Lipids, and Pesticides. One-way ANOVA indicated that the relative concentration of tylosin differed significantly across all the growth periods, except for the late-logarithmic and stabilization stages. This study provides important basic information on the secondary metabolite research of sf106, which will help us to understand and apply Streptomyces fradiae sf106 more comprehensively.

ABSTRACT Phytochemicals are promising antibacterials for the development of novel antibiofilm drugs, but their antibiofilm activity in physiologically relevant model systems is poorly characterized. As the host microenvironment can interfere with the activity of the phytochemicals, mimicking the complex environment found in biofilm associated infections is essential to predict the clinical potential of novel phytochemical-based antimicrobials. In the present study, we examined the antibiofilm activity of borneol, citral, and combinations of both as well as their Pickering emulsions against Staphylococcus aureus and Pseudomonas aeruginosa in an in vivo-like synthetic cystic fibrosis medium (SCFM2) model, an in vitro wound model (consisting of an artificial dermis and blood components at physiological levels), and an in vivo Galleria mellonella model. The Pickering emulsions demonstrated an enhanced biofilm inhibitory activity compared to both citral and the borneol/citral combination, reducing the minimum biofilm inhibitory concentration (MBIC) values up to 2 to 4 times against P. aeruginosa PAO1 and 2 to 8 times against S. aureus P8-AE1 in SCMF2. In addition, citral, the combination borneol/citral, and their Pickering emulsions can completely eliminate the established biofilm of S. aureus P8-AE1. The effectiveness of Pickering emulsions was also demonstrated in the wound model with a reduction of up to 4.8 log units in biofilm formation by S. aureus Mu50. Furthermore, citral and Pickering emulsions exhibited a significant degree of protection against S. aureus infection in the G. mellonella model. The present findings reveal the potential of citral- or borneol/citral-based Pickering emulsions as a type of alternative antibiofilm candidate to control pathogenicity in chronic infection. IMPORTANCE There is clearly an urgent need for novel formulations with antimicrobial and antibiofilm activity, but while there are plenty of studies investigating them using simple in vitro systems, there is a lack of studies in which (combinations of) phytochemicals are evaluated in relevant models that closely resemble the in vivo situation. Here, we examined the antibiofilm activity of borneol, citral, and their combination as well as Pickering emulsions (stabilized by solid particles) of these compounds. Activity was tested against Staphylococcus aureus and Pseudomonas aeruginosa in in vitro models mimicking cystic fibrosis sputum and wounds as well as in an in vivo Galleria mellonella model. The Pickering emulsions showed drastically increased antibiofilm activity compared to that of the compounds as such in both in vitro models and protected G. mellonella larvae from S. aureus-induced killing. Our data show that Pickering emulsions from phytochemicals are potentially useful for treating specific biofilm-related chronic infections.

For a long time, the development of the Lycium barbarum industry has been seriously restricted by root rot disease. In general, the occurrence of plant root rot is considered to be closely related to the composition and diversity of the soil microbial community. It is critical to understand the relationship between the occurrence of root rot in L. barbarum and the soil microbial composition. In this study, samples of the rhizosphere, rhizoplane, and root zone were collected from diseased and healthy plants. The V3–V4 region of bacterial 16S rDNA and the fungal ITS1 fragment of the collected samples were sequenced using Illumina MiSeq high-throughput sequencing technology. The sequencing results were first quality controlled and then aligned with the relevant databases for annotation and analysis. The richness of fungal communities in the rhizoplane and root zone of the healthy plants was significantly higher than that of the diseased plants (p < 0.05), and the community evenness and diversity of all the rhizoplane samples were significantly different from those of the rhizosphere and root zone. The richness of the bacterial communities in the rhizosphere and root zone of healthy plants was significantly greater than those of diseased plants (p < 0.05). The community composition of the rhizoplane was quite different from the other parts. The abundance of Fusarium in the rhizoplane and rhizosphere soil of diseased plants was higher than that in the corresponding parts of healthy plants. The abundances of Mortierella and Ilyonectria in the three parts of the healthy plants were correspondingly higher than those in the three parts of the diseased plants, and Plectosphaerella was the most abundant in the rhizoplane of diseased plants. There was little difference in the composition of the dominant bacteria at the phylum and genus levels between healthy plants and diseased plants, but the abundances of these dominant bacteria were different between healthy and diseased plants. Functional prediction showed that the bacterial community had the largest proportion of functional abundance belonging to metabolism. The functional abundances of the diseased plants, such as metabolism and genetic information processing, were lower than those of the healthy plants. The fungal community function prediction showed that the Animal Pathogen-Endophyte-Lichen Parasite-Plant Pathogen-Soil Saprotroph-Wood Saprotroph group had the largest functional abundance, and the corresponding fungi were Fusarium. In this study, we mainly discussed the differences in the soil microbial communities and their functions between the healthy and diseased L. barbarum cv. Ningqi-5, and predicted the functional composition of the microbial community, which is of great significance to understanding the root rot of L. barbarum.

Due to the complexity and similarity of plant leaves, it is very important to study an effective leaf-feature extraction method to improve the recognition rate of plant leaves. We study five multi-scale triangle representations: the triangle unsigned area representation (TUA), the triangle vertex angle representation (TVA) and three new representations, which we define as the gray average (TGA), the gray standard deviation (TGSD) and the side length integral (TSLI) on the triangle. In this method the curvature features of the contour, the texture features and the shape area feature are extracted to provide a multiscale leaf-feature description, and a new adaptive KNN for optimization method is proposed to improve the retrieval rate of leaf datasets. Experiments show that compared with the state-of-the-art methods, our method has higher accuracy on the Swedish and Flavia plant leaf datasets, which are respectively 99.35% and 99.43% with 84.76% Mean Average Precision (MAP) value and has comparable results on MPEG-7, kimia99 and kimia216 datasets. When our method is combined with KNN for optimization, the retrieval rate of the above datasets has been significantly improved, especially MAP on the Flavia dataset increases to 94.48%.

X-ray pulsar-based navigation (XPNAV) is one of the perfect ways for autonomous deep-space navigation in the future. Due to spacecraft state model errors and strong cosmic background noises, low navigation accuracy is one of the main problems in XPNAV. This paper proposes a robust navigation filtering method to reduce the serious effect of spacecraft state model errors and strong noises on XPNAV. This method uses state model errors and pulsar observation errors to estimate and correct the state model. And then, to predict the spacecraft' state in the next moment with high precision, the gain matrix is adjusted in quasi real-time by using the fading factor to ensure a minimized state estimation error variance in the next moment and an orthogonal residual sequence at different times. Finally, experimental results of multi-group simulations show that the proposed method had significantly improved navigation accuracy. And the accuracy of the proposed method is better than that of H∞ robust filter and STUKF, especially when the state model errors and noise are great. Under the same conditions, compared with the other two methods, the proposed method has the minimum navigation filtering error and the strongest robustness.

Staphylococcus aureus is one of the representative foodborne pathogens which forms biofilm. Antibiotics are widely applied in livestock husbandry to maintain animal health and productivity, thus contribute to the dissemination of antimicrobial resistant livestock and human pathogens, and pose a significant public health threat. Effect of antibiotic pressure on S. aureus biofilm formation, as well as the mechanism, remains unclear. In this study, the regulatory mechanism of low concentration of ampicillin on S. aureus biofilm formation was elucidated. The viability and biomass of biofilm with and without 1/4 MIC ampicillin treatment for 8 h were determined by XTT and crystal violet straining assays, respectively. Transcriptomics analysis on ampicillin-induced and non-ampicillin-induced biofilms were performed by RNA-sequencing, differentially expressed genes identification and annotation, GO functional and KEGG pathway enrichment. The viability and biomass of ampicillin-induced biofilm showed dramatical increase compared to the non-ampicillin-induced biofilm. A total of 530 differentially expressed genes (DEGs) with 167 and 363 genes showing up- and down-regulation, respectively, were obtained. Upon GO functional enrichment, 183, 252, and 21 specific GO terms in biological process, molecular function and cellular component were identified, respectively. Eight KEGG pathways including “Microbial metabolism in diverse environments”, “S. aureus infection”, and “Monobactam biosynthesis” were significantly enriched. In addition, “beta-lactam resistance” pathway was also highly enriched. In ampicillin-induced biofilm, the significant up-regulation of genes encoding multidrug resistance efflux pump AbcA, penicillin binding proteins PBP1, PBP1a/2, and PBP3, and antimicrobial resistance proteins VraF, VraG, Dlt, and Aur indicated the positive response of S. aureus to ampicillin. The up-regulation of genes encoding surface proteins ClfB, IsdA, and SasG and genes (cap5B and cap5C) which promote the adhesion of S. aureus in ampicillin induced biofilm might explain the enhanced biofilm viability and biomass.

Lactobacillus brevis is a major hop-resistance bacterium which poses significant challenge for the brewing industry, mainly due to the difficulty or incapability in detection by routine culturing methodology and its beer spoilage ability.This study aimed at investigating the VBNC state of a hop-resistance strain, L. brevis BM-LB13908. The culturable, total and viable numbers of L. brevis cells were calculated by MRS agar plate counting, acridine orange direct count (AODC) method and Live/Dead BacLight bacterial viability kit with fluorescence microscope. VBNC formation was induced by 189 ± 5.7 days under low-temperature storage or 27 ± 1.2 subcultures by continuous passage in beer, and VBNC cells induced by both strategies were recovered by adding catalase. In addition, insignificant difference in beer-spoilage ability was found in 3 states of L. brevis, including logarithmic growing, VBNC and recovered cells. This is the first study on the formation of VBNC state for L. brevis and beer-spoilage ability of both VBNC and recovered cells, which indicate L. brevis strain could cause beer spoilage without being detected by routine methodologies. The results derived from this study may support further study on L. brevis and other hop-resistance bacteria, and guidance on beer spoilage prevention and control, such as improvement for brewers on the microbiological quality control by using the improved culture method with catalase supplementation.

This study was to investigate whether natural borneol (NB) could enhance the anti-cancer effect of demethoxycurcumin (DCur) on HepG2 cell line by MTT assay, flow cytometry, and western blotting assay. Our results demonstrated that NB/DCur resulted in a significant decrease in cell viability due to pretreatment of NB enhancing the cellular uptake of DCur. Flow cytometric assay showed that NB/DCur-induced HepG2 cells growth inhibition was mainly caused by induction of G2/M arrest, as evidenced by accumulation of the G2/M cell population. Immunoblotting assay demonstrated that NB/DCur down-regulated expression levels of cdc2 and cyclin B1, which contributed to G2/M arrest. Moreover, NB/DCur elevated the level of intracellular reactive oxygen species (ROS), indicating that NB/DCur-induced G2/M arrest was achieved by triggered ROS-mediated DNA damage involving MAPK and Akt signaling pathways. Taken together, our results suggested that the combination of NB and DCur induced G2/M phase arrest in HepG2 through ROS overproduction. This study demonstrated that NB had the potential to be further developed into a chemosensitizer of DCur in the treatment of human cancers.

Naringin palmitate is an acylated product of naringin with palmitic acid, which is an effective substance for enhancing naringin lipophilicity. Lysozyme has strong ability to carry drugs and functional compounds in mammals. Herein, the ability of lysozyme to carry naringin palmitate was investigated by fluorescence and UV spectroscopy, and the differences in affinities of naringin and naringin palmitate for lysozyme were compared. Naringin palmitate was found to quench the fluorescence of lysozyme via static quenching. The binding constant and number of binding sites of naringin palmitate–lysozyme were more sensitive to high temperature than naringin–lysozyme. van der Waals forces and hydrogen bond formation played a major role in stabilizing naringin palmitate–lysozyme. No significant change of the structure and microenvironment of lyzozyme were obtained by addition of naringin palmitate. Thus, transport of naringin palmitate by lysozyme was verified. These findings are of importance and are attractive for the practical applications of naringin palmitate.

The aims of this study were to prepare a thermosensitive composite hydrogel (TCH) by mixing 24% (w/v) poloxamer 407 (P407), 16% (w/v) poloxamer 188 (P188) and 0.1% (w/v) carbopol 940 (C940), and to determine the effect of natural borneol/ (2-hydroxypropyl)-β-cyclodextrin (NB/HP-β-CD) inclusion complex on the phase transition temperature, mechanical, rheological properties, and release behaviors of the TCH using the tube inversion method, a texture analyzer, a rheometer, and in vitro release , respectively. The results showed that as the concentration of NB/HP-β-CD increased, the phase transition temperature of the TCH was increased from 37.26 to 38.34 °C and the mechanical properties of the TCH showed that the hardness, cohesiveness, strength, and adhesiveness were increased from 0.025 to 0.064 kg, 0.022 to 0.064 kg, 0.110 to 0.307 kg and 0.036 to 0.105 kg, respectively, but the rheological properties of the TCH showed that G′, G′′ and η were decreased from 7,760 to 157.50 Pa, 1,274 to 36.28 Pa and 1,252 to 25.37 Pas, respectively. The in vitro release showed that an increasing NB/HP-β-CD concentration decreased the release rate of NB from the TCH, but the amount of NB released was more than 96% at 60 min, which showed the TCH had good release behavior.

Thermal behavior of palm stearin (PS) and palm olein (PO) was explored by monitoring peak temperature transitions by differential scanning calorimetry (DSC). The fatty acid composition (FAC), isothermal crystallization kinetics studied by pulsed Nuclear Magnetic Resonance (pNMR) and isothermal microstructure were also compared. The results indicated that the fatty acid composition had an important influence on the crystallization process. PS and PO both exhibited more multiple endotherms than exotherms which showed irregular peak shapes. An increasing in cooling rate, generally, was associated with an increase in peak size. Application of the Avaimi equation to isothermal crystallization of PS and PO revealed different nucleation and growth mechanisms based on the Avrami exponents. PS quickly reached the end of crystallization because of more saturated triacylglycerol (TAG). The Avrami index of PS were the same as PO under the same isothermal condition at lower temperatrue, indicating that the crystallization mechanism of the two samples based on super-cooling state were the same. According to the polarized light microscope (PLM) images, crystal morphology of PS and PO was different. With the temperature increased, the structure of crystal network of both PS and PO gradually loosened.

Nε-(carboxymethyl) lysine (CML) is the most abundant advanced glycation end product (AGE), and frequently selected as an AGEs marker in laboratory studies. In this paper, the formation and inhibition of Nε-(carboxymethyl)lysine in saccharide-lysine model systems during microwave heating have been studied. The microwave heating treatment significantly promoted the formation of CML during Maillard reactions, which was related to the reaction temperature, time and type of saccharide. The order of CML formation for different saccharides was lactose > glucose > sucrose. Then, the inhibition effect on CML by five inhibitors was further examined. According to the results, ascorbic acid and tocopherol did not affect inhibition of CML, in contrast, thiamin, rutin and quercetin inhibited CML formation, and the inhibitory effects were concentration dependent.

Bladder cancer has become the most common malignant urinary carcinoma. Studies have shown that significant antioxidant and bladder cancer-fighting properties of several plant-based diets like Psidium guajava, ginger and amomum, are associated with their high kaempferol content. In this paper, we evaluated the antioxidant and anticancer activities of kaempferol and its mechanism of induction to apoptosis on bladder cancer cells. Our findings demonstrated that kaempferol showed an obvious radical scavenging activity in erythrocytes damaged by oxygen. Kaempferol promoted antioxidant enzymes, inhibited ROS generation and lipid peroxidation and finally prevented the occurrence of hemolysis. Additionally, kaempferol exhibited a strong inhibitory effect on bladder cancer cells and high safety on normal bladder cells. At the molecular level, kaempferol suppressed EJ bladder cancer cell proliferation by inhibiting the function of phosphorylated AKT (p-AKT), CyclinD1, CDK4, Bid, Mcl-1 and Bcl-xL, and promoting p-BRCA1, p-ATM, p53, p21, p38, Bax and Bid expression, and finally triggering apoptosis and S phase arrest. We found that Kaempferol exhibited strong anti-oxidant activity on erythrocyte and inhibitory effects on the growth of cancerous bladder cells through inducing apoptosis and S phase arrest. These findings suggested that kaempferol might be regarded as a bioactive food ingredient to prevent oxidative damage and treat bladder cancer.

This study was to investigate the synergistic effect of NB/Cur on growth and apoptosis in A375 human melanoma cell line by MTT assay, flow cytometry and Western blotting. Our results demonstrated that NB effectively synergized with Cur to enhance its antiproliferative activity on A375 human melanoma cells by induction of apoptosis, as evidenced by an increase in sub-G1 cell population, DNA fragmentation, PARP cleavage and caspase activation. Further mechanistic studies by Western blotting showed that after treatment of the cells with NB/Cur, up-regulation of the expression level of phosphorylated JNK and down-regulation of the expression level of phosphorylated ERK and Akt contributed to A375 cells apoptosis. Moreover, NB also potentiated Cur to trigger intracellular ROS overproduction and the DNA damage with up-regulation of the expression level of phosphorylated ATM, phosphorylated Brca1 and phosphorylated p53. The results indicate the combinational application potential of NB and Cur in treatments of cancers.

BackgroundBovine serum albumin (BSA) contains high affinity binding sites for several endogenous and exogenous compounds and has been used to replace human serum albumin (HSA), as these two compounds share a similar structure. Naringin palmitate is a modified product of naringin that is produced by an acylation reaction with palmitic acid, which is considered to be an effective substance for enhancing naringin lipophilicity. In this study, the interaction of naringin palmitate with BSA was characterised by spectroscopic and molecular docking techniques.Methodology/principal findingsThe goal of this study was to investigate the interactions between naringin palmitate and BSA under physiological conditions, and differences in naringin and naringin palmitate affinities for BSA were further compared and analysed. The formation of naringin palmitate-BSA was revealed by fluorescence quenching, and the Stern-Volmer quenching constant (KSV ) was found to decrease with increasing temperature, suggesting that a static quenching mechanism was involved. The changes in enthalpy (ΔH) and entropy (ΔS) for the interaction were detected at -4.11 ± 0.18 kJ·mol(-1) and -76.59 ± 0.32 J·mol(-1)·K(-1), respectively, which indicated that the naringin palmitate-BSA interaction occurred mainly through van der Waals forces and hydrogen bond formation. The negative free energy change (ΔG) values of naringin palmitate at different temperatures suggested a spontaneous interaction. Circular dichroism studies revealed that the α-helical content of BSA decreased after interacting with naringin palmitate. Displacement studies suggested that naringin palmitate was partially bound to site I (subdomain IIA) of the BSA, which was also substantiated by the molecular docking studies.Conclusions/significanceIn conclusion, naringin palmitate was transported by BSA and was easily removed afterwards. As a consequence, an extension of naringin applications for use in food, cosmetic and medicinal preparations may be clinically and practically significant, especially in the design of new naringin palmitate-inspired drugs.

Selenocystine (SeC) has been identified as a novel compound with broad-spectrum anticancer activities. Natural borneol (NB) is a monoterpenoid compound that has been used as a promoter of drug absorption. In the present study, we demonstrated that NB significantly enhanced the cellular uptake of SeC and potentiated its antiproliferative activity on HepG2 cells by induction of apoptosis. NB effectively synergized with SeC to reduce cancer cell growth through the triggering apoptotic cell death. Further mechanistic studies by Western blotting showed that treatment of the cells with NB and SeC activated the intrinsic apoptotic pathway by regulation of pro-survival and pro-apoptotic Bcl-2 family proteins. Treatment of the cells with NB and SeC induced the activation of p38MAPK and inactivation of Akt and ERK. NB also potentiated SeC to trigger intracellular ROS generation and DNA strand breaks as examined by Comet assay. Moreover, the thiol-reducing antioxidants effectively blocked the occurrence of cell apoptosis, which confirmed the important role of ROS in cell apoptosis. Taken together, these results reveal that NB strongly potentiates SeC-induced apoptosis in cancer cells by enhancement of cellular uptake and activation of ROS-mediated DNA damage. NB could be further developed as a chemosensitizer of SeC in treatment of human cancers.