Impairment of ovarian follicular development caused by titanium dioxide nanoparticles exposure involved in the TGF-ß/BMP/Smad pathway.

Titanium dioxide nanoparticles (TiO2 NPs) have been shown to induce reproductive system damages in animals. To better underline how TiO2 NPs act in reproductive system, female mice were exposed to 2.5, 5, or 10 mg/kg TiO2 NPs by gavage administration for 60 days, the ovary injuries, follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels as well as ovarian follicular development-related molecule expression were investigated. The results showed that TiO2 NPs exposure resulted in reduction of ovary weight and inhibition of ovarian follicular development. Furthermore, the suppression of follicular development was demonstrated to be closely related to higher FSH and LH levels, and higher expression of activin, follistatin, BMP2, BMP4, TGF-ß1, Smad2, Smad3, and Smad4 as well as decreased inhibin-α expression in mouse ovary in a dose-dependent manner. It implies that the impairment of ovarian follicular development caused by TiO2 NPs exposure may be mediated by TGF-ß signal pathway.

An F-box protein attenuates fungal xylanase-triggered immunity by destabilizing LRR-RLP NbEIX2 in a SOBIR1-dependent manner.

Receptor-like proteins (RLPs) lacking the cytoplasmic kinase domain play crucial roles in plant growth, development and immunity. However, what remains largely elusive is whether RLP protein levels are fine-tuned by E3 ubiquitin ligases, which are employed by receptor-like kinases for signaling attenuation. Nicotiana benthamiana NbEIX2 is a leucine-rich repeat RLP (LRR-RLP) that mediates fungal xylanase-triggered immunity. Here we show that NbEIX2 associates with an F-box protein NbPFB1, which promotes NbEIX2 degradation likely by forming an SCF E3 ubiquitin ligase complex, and negatively regulates NbEIX2-mediated immune responses. NbEIX2 undergoes ubiquitination and proteasomal degradation in planta. Interestingly, NbEIX2 without its cytoplasmic tail is still associated with and destabilized by NbPFB1. In addition, NbPFB1 also associates with and destabilizes NbSOBIR1, a co-receptor of LRR-RLPs, and fails to promote NbEIX2 degradation in the sobir1 mutant. Our findings reveal a distinct model of NbEIX2 degradation, in which an F-box protein destabilizes NbEIX2 indirectly in a SOBIR1-dependent manner.

Lensless light intensity model for quasi-spherical cell size measurement.

Quasi-spherical cell size measurement plays an important role in medical test. Traditional methods such as a microscope and a flow cytometer are either it depends on professionals and cannot be automated, or it is expensive and bulky, which are not suitable for point-of-care test. Lab-on-a-chip technology using the lensless imaging system gives a good solution for obtaining the quasi-spherical cell size. The diffraction effects and the low resolution are the two main problems faced by the lensless imaging system. In this paper, a lensless light intensity model for the quasi-spherical cell size measurement is given. First, the diffraction characteristics of a quasi-spherical cell edge are given. Then, a diffraction model at an arc edge is constructed based on the Fresnel diffraction at a straight edge. Using the diffraction model at an arc edge, we explained the mechanism of the formation of the quasi-spherical cell diffraction fringes. Finally, the light intensity of the first bright ring of the quasi-spherical cell diffraction pattern is used to achieve quasi-spherical cell size measurement. The required equipment and the measurement methods are extremely simple, very suitable for point-of-care test. The experimental results show that the proposed model can realize the statistical measurement of the quasi-spherical cells and the classification of the quasi-spherical cells with a difference of 1 [Formula: see text].

Extraction and activity of chemical constituents from Houttuynia cordata Thunb by ultrasonic method.

Polyphenols and flavonoids are phytochemicals that have essential roles in human nutrition. In this regard, the contents of polyphenols and flavonoids in Houttuynia cordata Thunb and their antioxidant activities were evaluated in the current study. Two Houttuynia cordata materials with the same chromosome number and chemical type were used to comprehensively assess the contents of total phenols and flavonoids in different parts of H. cordata. These chemical components were extracted by the ultrasonic method. The results showed that the total phenols and antioxidant capacity of different parts of H. cordata were significantly different. The content of polyphenols in roots and stems was low, the antioxidant capacity was weak, the total phenols in flowers and leaves were high, and the antioxidant capacity was strong. Therefore, different parts of H. cordata had different pharmacological and food effects. The whole herb can be used as Chinese herbal medicine, and its young leaves and roots can be used as vegetables. Flavonoids are the main phenolic components, and total phenols are the main components of antioxidant activity. It can explain a very significant positive correlation between total phenols and flavonoids. Therefore, in the further breeding work of H. cordata, the procedure can be simplified by determining one of the above indexes to predict the varieties with high total phenolic and antioxidant activity.

The fecal microbiota of patients with pancreatic ductal adenocarcinoma and autoimmune pancreatitis characterized by metagenomic sequencing.

BACKGROUND: The fecal microbiota in pancreatic ductal adenocarcinoma (PDAC) and in autoimmune pancreatitis (AIP) patients remains largely unknown. We aimed to characterize the fecal microbiota in patients with PDAC and AIP, and explore the possibility of fecal microbial biomarkers for distinguishing PDAC and AIP. METHODS: 32 patients with PDAC, 32 patients with AIP and 32 age- and sex-matched healthy controls (HC) were recruited and the fecal microbiotas were analyzed through high-throughput metagenomic sequencing. Alterations of fecal short-chain fatty acids were measured using gas chromatographic method. RESULTS: Principal coordinate analysis (PCoA) revealed that microbial compositions differed significantly between PDAC and HC samples; whereas, AIP and HC individuals tended to cluster together. Significant reduction of phylum Firmicutes (especially butyrate-producing bacteria, including Eubacterium rectale, Faecalibacterium prausnitzii and Roseburia intestinalis) and significant increase of phylum Proteobacteria (especially Gammaproteobacteria) were observed only among PDAC samples. At species level, when compared with HC samples, we revealed 24 and 12 differently enriched bacteria in PDAC and AIP, respectively. Functional analysis showed a depletion of short-chain fatty acids synthesis associated KO modules (e.g. Wood-Ljungdahl pathway) and an increase of KO modules associated with bacterial virulence (e.g. type II general secretion pathway). Consistent with the downregulation of butyrate-producing bacteria, gas chromatographic analysis showed fecal butyrate content was significantly decreased in PDAC group. Eubacterium rectale, Eubacterium ventrisum and Odoribacter splanchnicus were among the most important biomarkers in distinguishing PDAC from HC and from AIP individuals. Receiver Operating Characteristic analysis showed areas under the curve of 90.74% (95% confidence interval [CI] 86.47-100%), 88.89% (95% CI 73.49-100%), and 76.54% (95% CI 52.5-100%) for PDAC/HC, PDAC/AIP and AIP/HC, respectively. CONCLUSIONS: In conclusion, alterations in fecal microbiota and butyrate of patients with PDAC suggest an underlying role of gut microbiota for the pathogenesis of PDAC. Fecal microbial and butyrate as potential biomarkers may facilitate to distinguish patients with PDAC from patients with AIP and HCs which worth further validation.

Fecal Fusobacterium nucleatum harbored virulence gene fadA are associated with ulcerative colitis and clinical outcomes.

OBJECT: Fusobacterium nucleatum (F.nucleatum), a gram-negative, obligately anaerobe of oral commensal，has been regarded as culprit of periodontal diseases previously and is being unveiled as possible pathogen of gastrointestinal disorders. The key virulence factor of F.nucleatum is FadA adhesin for binding and invading of the host's epithelial cells. Here, we detected fecal F.nucleatum and virulence gene fadA in patients with ulcerative colitis(UC) and evaluated the clinical relevance with UC. METHODS AND SUBJECTS: A total of 310 subjects were enrolled including 100 patients with UC, 70 healthy controls (HC), 70 patients with irritable bowel syndrome subtype diarrhea(IBS-D), and 70 colorectal cancer patients(CRC). Stool samples of UC patients compared with healthy controls as well as IBS-D and CRC patients were collected for Polymerase Chain Reaction(PCR) detection of F.nucleatum (based on 16s rRNA) and virulence gene fadA. RESULTS: The detection rate of 16s rRNA based PCR for F.nucleatum of UC patients(39/100, 39.00%) and CRC(26/70, 37.14%) patients are significantly higher than HC (12/70, 17.14%, P < 0.01) and IBS-D patients (14/70, 20.00%, P < 0.01). Moreover, 19 samples were detected fadA positive from 39 F.nucleatum positive samples of UC patients (19/39, 48.72%), which is significantly higher than HC(2/12, 16.66%, P < 0.05). There were 3 samples detected fadA positive from 14 F.nucleatum positive samples of IBS-D patients(3/14, 21.43%) and 13 out of 26(50.00%) of CRC patients, which were both no significant differences compared with UC patients(21.4% vs 48.72%, P > 0.05; 50.00% vs 48.72%, P > 0.05). For both F.nucleatum and fadA gene positive patients, there were no statistical significances between erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), white blood cells(WBC), and hemoglobin compared with negative patients(defined by either F.nucleatum or fadA negative, or both negative). However, it is worth noting that detection rate of F.nucleatum with virulence gene fadA in patients of severe ulcerative colitis was significantly higher than patients with mild and moderate colitis(28.89% vs 10.91%, P < 0.05). In addition, the fecal F.nucleatum and fadA gene positive patients were more likely to have pancolitis other than left-sided colitis(pancolitis/left-sided colitis: 26.92% vs 10.42%, P < 0.05). CONCLUSIONS: The presence of F.nucleatum and fadA gene increased in UC patients, especially in patients with severe colitis and pancolitis. Strains of F.nucleatum harbored virulence gene fadA are suggested to play a role in the pathogenesis of UC.

Measurement of red blood cell size based on a lensless imaging system.

The variability in the size of red blood cells (RBCs) is an important additional diagnostic parameter for diseases, which has been established as a part of the complete blood count (CBC). The CBC can be performed using an automated flow cytometer, but it is too bulky and expensive for point-of-care testing. A miniaturized lensless imaging system is a competitive modality for a CBC, that is small and inexpensive. There are two challenges in developing a lensless imaging system for taking the CBC, which make the measurement of the RBC size very difficult: the diffraction effect and the low resolution. In this paper, the RBC radius measurement is replaced with a diffraction ring radius measurement. The diffraction ring radius is much larger than the RBC radius. This feature can improve the imaging resolution. Based on Fresnel diffraction, the relationship between the radius of RBCs and the diffraction fringes is analyzed. Finally, a complete measurement algorithm for determining the RBC size based on the lensless imaging system is given, which can be used to measure the variability in the size of RBCs. In our experiment, the maximum error is less than 6.74%.

A small cysteine-rich protein from two kingdoms of microbes is recognized as a novel pathogen-associated molecular pattern.

Pathogen-associated molecular patterns (PAMPs) are conserved molecules that are crucial for normal life cycle of microorganisms. However, the diversity of microbial PAMPs is little known. During screening of cell-death-inducing factors from the necrotrophic fungus Valsa mali, we identified a novel PAMP VmE02 that is widely spread in oomycetes and fungi. Agrobacterium tumefaciens-mediated transient expression or infiltration of recombinant protein produced by Escherichia coli was performed to assay elicitor activity of the proteins tested. Virus-induced gene silencing in Nicotiana benthamiana was used to determine the components involved in VmE02-triggered cell death. The role of VmE02 in virulence and conidiation of V. mali were characterized by gene deletion and complementation. We found that VmE02, together with some of its homologues from both oomycete and fungal species, exhibited cell-death-inducing activity in N. benthamiana. VmE02-triggered cell death was shown to be dependent on BRI1-ASSOCIATED KINASE-1, SUPPRESSOR OF BIR1-1, HSP90 and SGT1 in N. benthamiana. Deletion of VmE02 in V. mali greatly attenuated pathogen conidiation but not virulence, and treatment of N. benthamiana with VmE02 enhances plant resistance to Sclerotinia sclerotiorum and Phytophthora capsici. We conclude that VmE02 is a novel cross-kingdom PAMP produced by several fungi and oomycetes.

A Systems Vaccinology Approach Reveals Temporal Transcriptomic Changes of Immune Responses to the Yellow Fever 17D Vaccine.

In this study, we used a systems vaccinology approach to identify temporal changes in immune response signatures to the yellow fever (YF)-17D vaccine, with the aim of comprehensively characterizing immune responses associated with protective immunity. We conducted a cohort study in which 21 healthy subjects in China were administered one dose of the YF-17D vaccine; PBMCs were collected at 0 h and then at 4 h and days 1, 2, 3, 5, 7, 14, 28, 84, and 168 postvaccination, and analyzed by transcriptional profiling and immunological assays. At 4 h postvaccination, genes associated with innate cell differentiation and cytokine pathways were dramatically downregulated, whereas receptor genes were upregulated, compared with their baseline levels at 0 h. Immune response pathways were primarily upregulated on days 5 and 7, accompanied by the upregulation of the transcriptional factors JUP, STAT1, and EIF2AK2. We also observed robust activation of innate immunity within 2 d postvaccination and a durable adaptive response, as assessed by transcriptional profiling. Coexpression network analysis indicated that lysosome activity and lymphocyte proliferation were associated with dendritic cell (DC) and CD4+ T cell responses; FGL2, NFAM1, CCR1, and TNFSF13B were involved in these associations. Moreover, individuals who were baseline-seropositive for Abs against another flavivirus exhibited significantly impaired DC, NK cell, and T cell function in response to YF-17D vaccination. Overall, our findings indicate that YF-17D vaccination induces a prompt innate immune response and DC activation, a robust Ag-specific T cell response, and a persistent B cell/memory B cell response.

A Quantized CNN-Based Microfluidic Lensless-Sensing Mobile Blood-Acquisition and Analysis System.

This paper proposes a microfluidic lensless-sensing mobile blood-acquisition and analysis system. For a better tradeoff between accuracy and hardware cost, an integer-only quantization algorithm is proposed. Compared with floating-point inference, the proposed quantization algorithm makes a tradeoff that enables miniaturization while maintaining high accuracy. The quantization algorithm allows the convolutional neural network (CNN) inference to be carried out using integer arithmetic and facilitates hardware implementation with area and power savings. A dual configuration register group structure is also proposed to reduce the interval idle time between every neural network layer in order to improve the CNN processing efficiency. We designed a CNN accelerator architecture for the integer-only quantization algorithm and the dual configuration register group and implemented them in field-programmable gate arrays (FPGA). A microfluidic chip and mobile lensless sensing cell image acquisition device were also developed, then combined with the CNN accelerator to build the mobile lensless microfluidic blood image-acquisition and analysis prototype system. We applied the cell segmentation and cell classification CNN in the system and the classification accuracy reached 98.44%. Compared with the floating-point method, the accuracy dropped by only 0.56%, but the area decreased by 45%. When the system is implemented with the maximum frequency of 100 MHz in the FPGA, a classification speed of 17.9 frames per second (fps) can be obtained. The results show that the quantized CNN microfluidic lensless-sensing blood-acquisition and analysis system fully meets the needs of current portable medical devices, and is conducive to promoting the transformation of artificial intelligence (AI)-based blood cell acquisition and analysis work from large servers to portable cell analysis devices, facilitating rapid early analysis of diseases.

Effects of Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis on Physicochemical Characteristics and Immune Activity In Vitro of Hericium erinaceus Polysaccharides.

The polysaccharide is the main active substance contained in Hericium erinaceus and is commonly used in the treatment of neurasthenia, tumors, and digestive diseases. Six intracellular polysaccharide components were obtained from H. erinaceus fruiting bodies cultivated by ARTP (atmospheric and room temperature plasma) mutagenic strain (321) and the original strain (0605), respectively. This study was designed to investigate the physicochemical characteristics of these polysaccharide components and their potential immunomodulatory activities on RAW264.7 macrophages. The results showed that the yield of fruiting body cultivated by mutated strain increased by 22% and the polysaccharide content improved by 16% compared with the original one owing to ARTP mutagenesis. The molecular weight distribution and the monosaccharide compositions of polysaccharide components from H. erinaceus induced by ARTP mutagenesis were significantly different from that of the original one. The NO, IL-6, IL-10, IL-1ß, and TNF-α production activities of macrophages were enhanced by stimulation of 20% ethanol precipitated polysaccharides from H. erinaceus induced by ARTP mutagenesis. These results indicated that ARTP is an efficient and practical method for high polysaccharide content breeding of the H. erinaceus strain and this provided a reference for obtaining high quality resources and healthy product development from H. erinaceus.

Improved fruiting of the straw mushroom (Volvariella volvacea) on cotton waste supplemented with sodium acetate.

Compared with controls, treatment of the cultivation substrate for Volvariella volvacea with 0.02% NaAc at the "spraying water" stage increased the number of fruiting body primordia by 280, the mushroom yield by 16.25%, the number of fruiting bodies by 35.57%, and the biological efficiency by 16.28%. The average single mushroom weight increased by 19.33%, but there was no significant difference between treatments and controls. A correlation analysis revealed a significant (p < 0.05) positive correlation between the total yield and the number of fruiting bodies. Comparisons of the cost and profit values for the sodium acetate-treated and untreated groups revealed that the former generated a higher income for the grower. Our data indicate that sodium acetate treatment is a promising new method for increasing V. volvacea yields. A possible mechanism whereby sodium acetate increased the mushroom yield is discussed.

Saccharothrix yanglingensis Strain Hhs.015 Is a Promising Biocontrol Agent on Apple Valsa Canker.

Apple Valsa canker (AVC), caused by Valsa mali, results in a serious and persistent disease problem for apple production in China and is difficult to control by chemical and agricultural measures. In this study, we determined the inhibitory effects of an endophytic actinomycete Saccharothrix yanglingensis strain Hhs.015 on V. mali under laboratory and the field conditions. Fermentation broth (FB) of Hhs.015 significantly inhibited conidial germination and mycelial growth, causing malformed and dysfunctional fungal structures. Detached apple (Malus domestica 'Fuji') twigs smeared with FB before V. mali inoculation significantly reduced lesion development, especially sporulation. In the orchard trials conducted from 2010 to 2013, trunks and branches smeared with FB three times significantly reduced the number of new lesions. In addition, wounds after lesion surgery coated with FB also enhanced callus formation and significantly improved the cure rate. The results showed that S. yanglingensis Hhs.015 is effective against AVC in orchards. Thus, S. yanglingensis Hhs.015 can be further developed as an effective biocontrol agent for more sustainable management of AVC.

Within-season yield prediction with different nitrogen inputs under rain-fed condition using CERES-Wheat model in the northwest of China.

BACKGROUND: Yield prediction within season is of great use to improve agricultural risk management and decision making. The objectives of this study were to access the yield forecast performance with increasing nitrogen inputs and to determine when the acceptable predicted yield can be achieved using the CERES-Wheat model. RESULTS: the calibrated model simulated wheat yield very well under various water and nitrogen conditions. Long-term simulation demonstrated that nitrogen input enlarged the annual variability of wheat yield generally. Within-season yield prediction showed that, regardless of nitrogen inputs, yield forecasts in the later growing season improved the accuracy and reduced the uncertainty of yield prediction. In a low-yielding year (2011-2012) and a high-yielding year (1991-1992), the date of acceptable predicted yield was achieved 62 and 65 days prior to wheat maturity, respectively. In a normal-yielding year (1983-1984), inadequate precipitation after the jointing stage in most historical years led to the underestimation of wheat yield and the date of accurate yield prediction was delayed to 235-250 days after simulation (7-22 days prior to maturity) for different N inputs. CONCLUSION: Yield prediction was highly influenced by the distribution of meteorological elements during the growing season and may show great improvement if future weather can be reliably forecast early. © 2015 Society of Chemical Industry.

[Function of nonribosomal peptide synthetase gene VmNRPS12 of Valsa mali].

Objective: Nonribosomal peptide synthetase (NRPS) plays an important role in plant-pathogenic fungi interaction. In this study, we analyzed the role of VmNRPS12 during Valsa mali (V. mali) infection and pathogenecity, which may shed new insights into the pathogenesis of V. mali. Methods: Based on the genome of V. mali, we obtained one NPRS, designated as VmNRPS12. The expression profile of VmNRPS12 was analyzed by qRT-PCR. Through Double-joint PCR and PEG-mediated protoplast transformation, VmNRPS12-knockout mutants were generated. PCR and Southern blot hybridization were applied to verify the positive mutant. Then genetic complementation was performed by transforming mutant protoplast with VmNRPS12 original gene. Finally, vegetative growth, conidiation and pathogenicity were examined. Results: qRT-PCR analyses showed that VmNRPS12 was upregulated during the early infection stages of V. mali, and expressed remarkably at 48 hours post inoculation (138.6-fold). Compared with the wild type 03-8, VmNRPS12-knockout mutant showed no obvious change in vegetative growth and conidiation on PDA medium. Notably, the VmNRPS12-knockout mutant exhibited significantly reduced virulence on apple twigs. Furthermore, complementation of VmNRPS12 restored the pathogenecity of the VmNRPS12-knockout mutant approximately to the wild type 03-8. Conclusion: VmNRPS12 contributes positively to the pathogenicity of V. mali.

Genome sequence of Valsa canker pathogens uncovers a potential adaptation of colonization of woody bark.

Canker caused by ascomycetous Valsa species are among the most destructive diseases of woody plants worldwide. These pathogens are distinct from other pathogens because they only effectively attack tree bark in the field. To unravel the potential adaptation mechanism of bark colonization, we examined the genomes of Valsa mali and Valsa pyri that preferentially infect apple and pear, respectively. We reported the 44.7 and 35.7 Mb genomes of V. mali and V. pyri, respectively. We also identified the potential genomic determinants of wood colonization by comparing them with related cereal pathogens. Both genomes encode a plethora of pathogenicity-related genes involved in plant cell wall degradation and secondary metabolite biosynthesis. In order to adapt to the nutrient limitation and low pH environment in bark, they seem to employ membrane transporters associated with nitrogen uptake and secrete proteases predominantly with acidic pH optima. Remarkably, both Valsa genomes are especially suited for pectin decomposition, but are limited in lignocellulose and cutin degradation. Besides many similarities, the two genomes show distinct variations in many secondary metabolism gene clusters. Our results show a potential adaptation of Valsa canker pathogens to colonize woody bark. Secondary metabolism gene clusters are probably responsible for this host specificity.

Prediction of serine/threonine phosphorylation sites in bacteria proteins.

As a critical post-translational modification, phosphorylation plays important roles in regulating various biological processes, while recent studies suggest that phosphorylation in bacteria is also critical for functional signaling transduction. Since identification of phosphorylation substrates and sites is fundamental for understanding the phosphorylation mediated regulatory mechanism, a number of studies have been contributed to this area. Since experimental identification of phosphorylation sites is time-consuming and labor-intensive, computational predictions attract much attention for its convenience to provide helpful information. However, although there are a large number of computational studies in eukaryotes, predictions in bacteria are still rare. In this study, we present a new predictor of cPhosBac to predict phosphorylation serine/threonine in bacteria proteins. The predictor is developed with CKSAAP algorithm, which was combined with motif length selection to optimize the prediction, which achieves promising performance. The online service of cPhosBac is available at: http://netalign.ustc.edu.cn/cphosbac/ .

Local feature acquisition and global context understanding network for very high-resolution land cover classification.

Very high-resolution remote sensing images hold promising applications in ground observation tasks, paving the way for highly competitive solutions using image processing techniques for land cover classification. To address the challenges faced by convolutional neural network (CNNs) in exploring contextual information in remote sensing image land cover classification and the limitations of vision transformer (ViT) series in effectively capturing local details and spatial information, we propose a local feature acquisition and global context understanding network (LFAGCU). Specifically, we design a multidimensional and multichannel convolutional module to construct a local feature extractor aimed at capturing local information and spatial relationships within images. Simultaneously, we introduce a global feature learning module that utilizes multiple sets of multi-head attention mechanisms for modeling global semantic information, abstracting the overall feature representation of remote sensing images. Validation, comparative analyses, and ablation experiments conducted on three different scales of publicly available datasets demonstrate the effectiveness and generalization capability of the LFAGCU method. Results show its effectiveness in locating category attribute information related to remote sensing areas and its exceptional generalization capability. Code is available at https://github.com/lzp-lkd/LFAGCU .

Combined Peptidomics and Metabolomics Analyses to Characterize the Digestion Properties and Activity of Stropharia rugosoannulata Protein-Peptide-Based Materials.

Protein-peptide-based materials typically possess high nutritional value and various physiological regulatory activities. This study evaluated the digestion, metabolism, and activity of Stropharia rugosoannulata protein-peptide-based materials. After the S. rugosoannulata protein-peptide-based materials were digested (simulated) orally, in the stomach, and in the intestines, the proportions of >10,000 Da, 5000~10,000 Da, and <180 Da in the digestion products increased, and the peptide content was maintained at more than 120 mg/g dry weight. The digestion products of eight test groups with different oral-gastrointestinal digestion-level settings all had suitable ACE inhibitory activity (IC50 range 0.004~0.096 mg/mL). The main metabolite groups were lipid-like molecules, fatty acids, carboxylic acids, their derivatives, amino acids, peptides, and analogs. Bile and glycosylated amino acids were the main compounds that caused differences between groups. KEGG pathways enriched in differentially expressed metabolites included eight significantly upregulated pathways, including valine, leucine, and isoleucine biosynthesis, etc., and six significantly downregulated pathways, including the citric acid cycle (tricarboxylic acid cycle), etc. The arginine and proline metabolism pathways and the aminoacyl-tRNA biosynthesis pathways were upregulation and downregulation pathways that enriched multiple differentially expressed metabolites. Twenty-six metabolites, including bile acids, total bile acids, and the essential amino acids L-isoleucine and L-leucine, were differentially expressed metabolite markers of the protein-peptide-based material oral-gastrointestinal digestion products.

Categorization of the effects of E. coli LF82 and mutants lacking the chuT and shuU genes on survival, the transcriptome, and metabolome in germ-free honeybee.

The precise etiology of inflammatory bowel diseases (IBDs) remains elusive. The Escherichia coli strain LF82 (LF82) is known to be associated with IBD, and we hypothesized that this association may be related to the chuT and shuU genes. Here we constructed a germ-free (GF) honeybee model to investigate the effects of LF82 chuT and shuU genes on the honeybee intestine and their mechanisms. The chuT and shuU gene deletion strains LF82∆chuT and LF82∆shuU were generated by CRISPR-Cas9. These strains, together with nonpathogenic E. coli MG1655 (MG1655) and wildtype LF82, were allowed to colonize the guts of GF honeybees to establish single bacterial colonization models. Intestinal permeability was assessed following the administration of a sterile Brilliant Blue (FCF) solution. Comprehensive transcriptomic and metabolomic analyses of intestinal samples indicated that MG1655 had few disadvantageous effects on honeybees. Conversely, colonization with LF82 and its gene-deletion mutants provoked pronounced activation of genes associated with innate immune pathways, stimulated defensive responses, and induced expression of genes associated with inflammation, oxidative stress, and glycosaminoglycan degradation. Crucially, the LF82∆chuT and LF82∆shuU strains perturbed host heme and iron regulation, as well as tryptophan metabolism. These findings suggest that the deletion of chuT and shuU genes in E. coli LF82 may alleviate intestinal inflammation by partially modulating tryptophan catabolism. Our study proposes that targeting iron uptake mechanisms could be a potential strategy to mitigate the virulence of IBD-associated bacteria.