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1.
Front Vet Sci ; 11: 1387853, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38835895

RESUMO

Locoweed is a poisonous plant widely present in grasslands around the world. Swainsonine (SW), an indole alkaloid that, is the main toxic component of the locoweed. To understand the mechanism of SW-induced toxicity and to delineate the metabolic profile of locoweed poisoning we performed the LC-MS/MS untargeted metabolomic study to analyze metabolites in SW-treated renal tubular epithelial cells (0.8 mg/mL, 12 h) and in order to identify the SW-induced metabolomic changes. The analysis identified 2,563 metabolites in positive ion mode and 1,990 metabolites in negative ion mode. Our results showed that the metabolites were mainly benzenoids, lipids and lipid-like molecules, nucleosides, nucleotides, and analogs, organic acids, and derivatives. The differential metabolites were primarily enriched in pathways involving bile secretion, primary bile acid biosynthesis, riboflavin metabolism, ferroptosis, drug metabolism-cytochrome P450, and primidine metabolism. We have screened out substances such as swainsonine, 3alpha,7alpha-Dihydroxy-5beta-cholestanate, 2-Hydroxyiminostilbene, and glycochenodeoxycholate, which may have the potential to serve as biomarkers for swainsonine poisoning. This study provides insights into the types of metabolomic alteration in renal tubular epithelial cells induced by swainsonine.

2.
Ecotoxicol Environ Saf ; 279: 116515, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38810283

RESUMO

Pyrrolizidine alkaloids (PAs) are a group of naturally occurring alkaloids widely present in plants. PAs are highly hepatotoxic and have been documented to cause many incidents of human and animal poisoning. Retrorsine (RTS) is a pyrrolizidine alkaloid (PA) derived from the Compositae Senecio, which has been shown to cause hepatotoxicity. Human liver poisoning occurs through the consumption of RTS-contaminated food, and animals can also be poisoned by ingesting RTS-containing toxic plants. The mechanism of RTS-induced liver toxicity is not fully understood. In this study, we demonstrated that RTS-induced oxidative stress plays a pivotal role in RTS-induced liver toxicity involving apoptosis and autophagy. The results showed that RTS treatment in the cultured Primary rat hepatocytes caused cytotoxicity and release of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in a time- and dose-dependent manner. Our study showed that treatment of RTS induced ROS and MDA (malondialdehyde, a lipid peroxidation marker) in the hepatocytes, and reduced antioxidant capacity (GSH content, SOD activity), suggesting RTS treatment caused oxidative stress response in the hepatocytes. Furthermore, we found that RTS induced apoptosis and autophagy in the hepatocytes, and RTS-induced apoptosis and autophagy could be alleviated by ROS scavenger N-acetylcysteine (NAC) and the MAPK pathway inhibitors suggesting ROS/MAPK signaling pathway plays a role in RTS induced apoptosis and autophagy. Collectively, this study reveals the regulatory mechanism of oxidative stress in RTS-induced apoptosis and autophagy in the hepatocytes, providing important insights of molecular mechanisms of hepatotoxicity induced by RTS and related pyrrolizidine alkaloids in liver. This mechanism provides a basis for the prevention and treatment of PA poisoning in humans and animals.


Assuntos
Apoptose , Autofagia , Hepatócitos , Estresse Oxidativo , Alcaloides de Pirrolizidina , Animais , Estresse Oxidativo/efeitos dos fármacos , Hepatócitos/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Alcaloides de Pirrolizidina/toxicidade , Ratos , Masculino , Espécies Reativas de Oxigênio/metabolismo , Ratos Sprague-Dawley , Células Cultivadas , Aspartato Aminotransferases , Alanina Transaminase
3.
Chem Biol Interact ; 394: 110990, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38579922

RESUMO

Swainsonine (SW) is the main toxic component of locoweed. Previous studies have shown that kidney damage is an early pathologic change in locoweed poisoning in animals. Trehalose induces autophagy and alleviates lysosomal damage, while its protective effect and mechanism against the toxic injury induced by SW is not clear. Based on the published literature, we hypothesize that transcription factor EB(TFEB) -regulated is targeted by SW and activating TFEB by trehalose would reverse the toxic effects. In this study, we investigate the mechanism of protective effects of trehalose using renal tubular epithelial cells. The results showed that SW induced an increase in the expression level of microtubule-associated protein light chain 3-II and p62 proteins and a decrease in the expression level of ATPase H+ transporting V1 Subunit A, Cathepsin B, Cathepsin D, lysosome-associated membrane protein 2 and TFEB proteins in renal tubular epithelial cells in a time and dose-dependent manner suggesting TFEB-regulated lysosomal pathway is adversely affected by SW. Conversely, treatment with trehalose, a known activator of TFEB promote TFEB nuclear translocation suggesting that TFEB plays an important role in protection against SW toxicity. We demonstrated in lysosome staining that SW reduced the number of lysosomes and increased the luminal pH, while trehalose could counteract these SW-induced effects. In summary, our results demonstrated for the first time that trehalose could alleviate the autophagy degradation disorder and lysosomal damage induced by SW. Our results provide an interesting method for reversion of SW-induced toxicity in farm animals and furthermore, activation of TFEB by trehalose suggesting novel mechanism of treating lysosomal storage diseases.


Assuntos
Autofagia , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos , Células Epiteliais , Túbulos Renais , Lisossomos , Swainsonina , Trealose , Animais , Autofagia/efeitos dos fármacos , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Linhagem Celular , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Túbulos Renais/efeitos dos fármacos , Túbulos Renais/patologia , Túbulos Renais/metabolismo , Túbulos Renais/citologia , Lisossomos/metabolismo , Lisossomos/efeitos dos fármacos , Swainsonina/toxicidade , Trealose/farmacologia
4.
Appl Microbiol Biotechnol ; 108(1): 314, 2024 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-38683435

RESUMO

The importance of dietary fiber (DF) in animal diets is increasing with the advancement of nutritional research. DF is fermented by gut microbiota to produce metabolites, which are important in improving intestinal health. This review is a systematic review of DF in pig nutrition using in vitro and in vivo models. The fermentation characteristics of DF and the metabolic mechanisms of its metabolites were summarized in an in vitro model, and it was pointed out that SCFAs and gases are the important metabolites connecting DF, gut microbiota, and intestinal health, and they play a key role in intestinal health. At the same time, some information about host-microbe interactions could have been improved through traditional animal in vivo models, and the most direct feedback on nutrients was generated, confirming the beneficial effects of DF on sow reproductive performance, piglet intestinal health, and growing pork quality. Finally, the advantages and disadvantages of different fermentation models were compared. In future studies, it is necessary to flexibly combine in vivo and in vitro fermentation models to profoundly investigate the mechanism of DF on the organism in order to promote the development of precision nutrition tools and to provide a scientific basis for the in-depth and rational utilization of DF in animal husbandry. KEY POINTS: • The fermentation characteristics of dietary fiber in vitro models were reviewed. • Metabolic pathways of metabolites and their roles in the intestine were reviewed. • The role of dietary fiber in pigs at different stages was reviewed.


Assuntos
Ração Animal , Fibras na Dieta , Fermentação , Microbioma Gastrointestinal , Animais , Fibras na Dieta/metabolismo , Suínos , Microbioma Gastrointestinal/fisiologia , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Ácidos Graxos Voláteis/metabolismo
5.
Med Phys ; 2024 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-38306473

RESUMO

BACKGROUND: Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) plays a crucial role in the diagnosis and measurement of hepatocellular carcinoma (HCC). The multi-modality information contained in the multi-phase images of DCE-MRI is important for improving segmentation. However, this remains a challenging task due to the heterogeneity of HCC, which may cause one HCC lesion to have varied imaging appearance in each phase of DCE-MRI. In particular, some phases exhibit inconsistent sizes and boundaries will result in a lack of correlation between modalities, and it may pose inaccurate segmentation results. PURPOSE: We aim to design a multi-modality segmentation model that can learn meaningful inter-phase correlation for achieving HCC segmentation. METHODS: In this study, we propose a two-stage progressive attention segmentation framework (TPA) for HCC based on the transformer and the decision-making process of radiologists. Specifically, the first stage aims to fuse features from multi-phase images to identify HCC and provide localization region. In the second stage, a multi-modality attention transformer module (MAT) is designed to focus on the features that can represent the actual size. RESULTS: We conduct training, validation, and test in a single-center dataset (386 cases), followed by external test on a batch of multi-center datasets (83 cases). Furthermore, we analyze a subgroup of data with weak inter-phase correlation in the test set. The proposed model achieves Dice coefficient of 0.822 and 0.772 in the internal and external test sets, respectively, and 0.829, 0.791 in the subgroup. The experimental results demonstrate that our model outperforms state-of-the-art models, particularly within subgroup. CONCLUSIONS: The proposed TPA provides best segmentation results, and utilizing clinical prior knowledge for network design is practical and feasible.

6.
Abdom Radiol (NY) ; 49(2): 471-483, 2024 02.
Artigo em Inglês | MEDLINE | ID: mdl-38200213

RESUMO

PURPOSE: The ideal contrast agent for imaging patients with hepatocellular carcinoma (HCC) following locoregional therapies (LRT) remains uncertain. We conducted a meta-analysis to assess the diagnostic performance of magnetic resonance imaging with extracellular contrast agent (ECA-MRI) and hepatobiliary agent (EOB-MRI) in detecting residual or recurrence HCC following LRT. METHODS: Original studies comparing the diagnostic performance of ECA-MRI and EOB-MRI were systematically identified through comprehensive searches in PubMed, EMBASE, Cochrane Library and Web of Science databases. The pooled sensitivity and specificity of ECA-MRI and EOB-MRI were calculated using a bivariate-random-effects model. Subgroup-analyses were conducted to compare the diagnostic performance of ECA-MRI and EOB-MRI according to different variables. Meta-regression analysis was employed to explore potential sources of study heterogeneity. RESULTS: A total of 15 eligible studies encompassing 803 patients and 1018 lesions were included. Comparative analysis revealed no significant difference between ECA-MRI and EOB-MRI in the overall pooled sensitivity (87% vs. 79%) and specificity (92% vs. 96%) for the detection of residual or recurrent HCC after LRT (P = 0.41), with comparable areas under the HSROC of 0.95 and 0.92. Subgroup analyses indicated no significant diagnostic performance differences between ECA-MRI and EOB-MRI according to study design, type of LRT, most common etiology of liver disease, baseline lesion size, time of post-treated examination and MRI field strength (All P > 0.05). CONCLUSION: ECA-MRI exhibited overall comparable diagnostic performance to EOB-MRI in assessing residual or recurrent HCC after LRT.


Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Carcinoma Hepatocelular/diagnóstico por imagem , Carcinoma Hepatocelular/terapia , Meios de Contraste , Neoplasias Hepáticas/diagnóstico por imagem , Neoplasias Hepáticas/terapia , Gadolínio DTPA , Imageamento por Ressonância Magnética/métodos , Sensibilidade e Especificidade , Estudos Retrospectivos
7.
Hortic Res ; 11(1): uhad249, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38288255

RESUMO

Vacuole largely dictates the fruit taste and flavor, as most of the sugars and organic acids are stored in the vacuoles of the fruit. However, difficulties associated with vacuole separation severely hinder identification and characterization of vacuolar proteins in fruit species. In this study, we established an effective approach for separating vacuoles and successfully purified vacuolar protein from six types of citrus fruit with varying patterns of sugar and organic acid contents. By using label-free LC-MS/MS proteomic analysis, 1443 core proteins were found to be associated with the essential functions of vacuole in citrus fruit. Correlation analysis of metabolite concentration with proteomic data revealed a transporter system for the accumulation of organic acid and soluble sugars in citrus. Furthermore, we characterized the physiological roles of selected key tonoplast transporters, ABCG15, Dict2.1, TMT2, and STP7 in the accumulation of citric acid and sugars. These findings provide a novel perspective and practical solution for investigating the transporters underlying the formation of citrus taste and flavor.

8.
Am J Cardiol ; 211: 209-218, 2024 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-37984642

RESUMO

To investigate the long-term effects of 2 commonly used low-osmolar contrast media, iohexol and iopromide, on renal function and survival in patients who underwent coronary angiography. A total of 14,141 cardiology patients from 2006 to 2013 were recruited, of whom 1,793 patients (679 patients on iohexol and 1,114 on iopromide) were evaluated for long-term renal impairment and 5,410 patients (1,679 patients on iohexol and 3,731 on iopromide) were admitted for survival analyses spanning as long as 15 years. Univariate and multivariate logistic regression were used to explore the risk factors for long-term renal impairment. Cox proportional hazard regression was used to investigate the risk factors affecting survival. Propensity score matching and inverse probability of treatment weighting were applied to balance the baseline clinical characteristics. Patients receiving iohexol demonstrated a greater occurrence of renal impairment compared with those who received iopromide. Such difference remained consistent both before and after propensity score matching or inverse probability of treatment weighting, with a statistical significance of p <0.05. Among clinical variables, receiving contrast-enhanced contrast tomography/magnetic resonance imaging during follow-up, antihypertensive medication usage, presence of proteinuria, and anemia were identified as risk factors for long-term renal impairment (p = 0.041, 0.049, 0.006, and 0.029, respectively). During survival analyses, the difference was insignificant after propensity score matching and inverse probability of treatment weighting. In conclusion, administration of iohexol was more likely to induce long-term renal impairment than iopromide, particularly among patients diagnosed with anemia and proteinuria and those taking antihypertensive medication and with additional contrast exposure. The all-cause mortality, however, showed no significant difference between iohexol and iopromide administration.


Assuntos
Anemia , Insuficiência Renal , Humanos , Iohexol/efeitos adversos , Angiografia Coronária/efeitos adversos , Angiografia Coronária/métodos , Meios de Contraste/efeitos adversos , Anti-Hipertensivos , Insuficiência Renal/induzido quimicamente , Insuficiência Renal/epidemiologia , Proteinúria/induzido quimicamente , Ácidos Tri-Iodobenzoicos/efeitos adversos
9.
Open Life Sci ; 18(1): 20220635, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37426617

RESUMO

Previous studies suggest that the senescence and death of the replaceable bud of the Chinese chestnut cultivar (cv.) "Tima Zhenzhu" involves programmed cell death (PCD). However, the molecular network regulating replaceable bud PCD is poorly characterized. Here, we performed transcriptomic profiling on the chestnut cv. "Tima Zhenzhu" replaceable bud before (S20), during (S25), and after (S30) PCD to unravel the molecular mechanism underlying the PCD process. A total of 5,779, 9,867, and 2,674 differentially expressed genes (DEGs) were discovered upon comparison of S20 vs S25, S20 vs S30, and S25 vs S30, respectively. Approximately 6,137 DEGs common to at least two comparisons were selected for gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses to interrogate the main corresponding biological functions and pathways. GO analysis showed that these common DEGs could be divided into three functional categories, including 15 cellular components, 14 molecular functions, and 19 biological processes. KEGG analysis found that "plant hormone signal transduction" included 93 DEGs. Overall, 441 DEGs were identified as related to the process of PCD. Most of these were found to be genes associated with ethylene signaling, as well as the initiation and execution of various PCD processes.

10.
Chin J Acad Radiol ; 6(1): 47-56, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36741827

RESUMO

Background: Acute respiratory distress syndrome (ARDS) is a critical disease in the intensive care unit (ICU) with high morbidity and mortality. The accuracy for predicting ARDS patients' outcome with mechanical ventilation is limited, and most based on clinical information. Methods: The patients diagnosed with ARDS between January 2014 and June 2019 were retrospectively recruited. Radiomics features were extracted from the upper, middle, and lower levels of the lung, and were further analyzed with the primary outcome (28-day mortality after ARDS onset). The univariate and multivariate logistic regression analyses were applied to figure out risk factors. Various predictive models were constructed and compared. Results: Of 366 ARDS patients recruited in this study, 276 (median age, 64 years [interquartile range, 54-75 years]; 208 male) survive on the Day 28. Among all factors, the APACHE II Score (OR 2.607, 95% CI 1.896-3.584, P < 0.001), the Radiomics_Score of the middle lung (OR 2.230, 95% CI 1.387-3.583, P = 0.01), the Radiomics_Score of the lower lung (OR 1.633, 95% CI 1.143-2.333, P = 0.01) were associated with the 28-day mortality. The clinical_radiomics predictive model (AUC 0.813, 95% CI 0.767-0.850) show the best performance compared with the clinical model (AUC 0.758, 95% CI 0.710-0.802), the radiomics model (AUC 0.692, 95% CI 0.641-0.739) and the various ventilator parameter-based models (highest AUC 0.773, 95% CI 0.726-0.815). Conclusions: The radiomics features of chest CT images have incremental values in predicting the 28-day mortality in ARDS patients with mechanical ventilation. Supplementary Information: The online version contains supplementary material available at 10.1007/s42058-023-00116-x.

11.
ACS Infect Dis ; 9(3): 497-506, 2023 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-36787534

RESUMO

With the continuous emergence and spread of drug-resistant and multi-drug-resistant Staphylococcus aureus, traditional antibiotic treatment has gradually lost its effect. There is an urgent need to develop and study new and effective bio-green inhibitors to control S. aureus. In this study, the S. aureus phage DZ25 was isolated from milk and the lysin LysDZ25 with excellent tolerance to serum and NaCl solution was identified. Subsequently, to improve the lytic activity and thermal stability of LysDZ25, RoseTTAFold was used to construct three-dimensional (3D) structures, molecular dynamics (MD) simulation was used for conformational acquisition, and the MDL strategy previously developed in our lab was used to rationally design variants. After two rounds of rational design, the optimal variant with improved thermal stability, S333V/N245R/D299L, was obtained, and its half-life time was 4.0-fold that of wild-type LysDZ25. At 37, 40, 45, and 50 °C, the lytic activity of the optimal triple-point variant S333V/N245R/D299L was increased by 17.3-, 26.7-, 20.2-, and 50.1-fold compared with that of the wild-type LysDZ25, respectively. Finally, cell count was used to evaluate the lytic activity, and the results showed that the optimal variant S333V/N245R/D299L could drop about 3.5 log 10 values compared with the control and about 2.6 log 10 values compared with the wild-type LysDZ25.


Assuntos
Staphylococcus aureus Resistente à Meticilina , Infecções Estafilocócicas , Humanos , Staphylococcus aureus , Fagos de Staphylococcus/genética , Antibacterianos/farmacologia
12.
Res Microbiol ; 174(3): 104011, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36455782

RESUMO

Agrobacterium fabrum is a phytopathogen that causes the crown gall disease. Some plant-derived molecules, e.g. phenols, directly affect A. fabrum-plant interactions. Here, we characterize a phenolic catabolism-related gene, atu1420, that affects the pathogenicity of A. fabrum. Atu1420 is predicted to be an O-demethylase with high structural homology to Sphingomonas paucimobilis LigM. The HPLC-UV analysis showed that atu1420 affected the degradation of acetosyringone (AS). The deletion of atu1420 gene significantly enhanced the AS-induced virulence (vir) gene expression. atu1420 was shown to relieve the inhibitory effect of vanillic acid on the AS-induced vir gene expression and the growth of A. fabrum. The expression of atu1420 and the degradation of AS in A. fabrum C58 was up-regulated by the addition of indole acetic acid (IAA). The inhibitory effect of IAA on the AS-induced vir gene expression was partially relieved by the deletion of atu1420 gene, indicating that accelerating the degradation of AS is one of the ways that IAA inhibits vir genes induction. Furthermore, atu1420 mutant produced more pronounced tumors on kalanchoe leaves than the wild-type strain. These findings reveal the role of atu1420 in A. fabrum-host interactions and will broaden our understanding of the regulatory network of the interactions.


Assuntos
Agrobacterium , Fenóis , Virulência/genética , Fenóis/farmacologia , Fenóis/metabolismo , Agrobacterium/genética , Agrobacterium/metabolismo , Fatores de Virulência/genética , Fatores de Virulência/metabolismo , Agrobacterium tumefaciens/genética , Regulação Bacteriana da Expressão Gênica , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo
13.
Toxicol Lett ; 373: 41-52, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36402259

RESUMO

The indolizidine alkaloid, swainsonine (SW), is the main toxic component of locoweed, which can cause locoism in animals with characteristic neurological dysfunction. Pathological manifestations at cellular level include extensive vacuolar degeneration. Studies have shown that SW can induces autophagy, but the role and mechanism of autophagy in SW-induced vacuolar degeneration is unclear. In this study, we analyzed the role of autophagy in SW-induced cell injury in mouse hippocampal neurons cell line (HT22) using western blotting, qRT-PCR, transmission electron microscopy and immunofluorescence microscopy. The results showed that the expressions of LC3-II, ATG5, Beclin1 and p62 proteins and their mRNAs in HT22 cells were induced by SW treatment. The SW treatment increased the number of autophagosomes with enhanced fluorescence intensity of monodansylcadaverine (MDC) and LC3-II in a time-dose dependent manner. The results of lysosome staining showed that SW could increase the number of lysosomes, increase the intraluminal pH. Transmission electron microscopy results indicate that SW induced autophagosomes, and Baf A1 could effectively alleviate SW-induced vacuolar degeneration. At the molecular level, SW treatment inhibited the expression of p-PI3K, p-AKT, p-ERK, p-AMPK, p-mTOR, p-p70S6K and p-4EBP1 and promoted the expression of p53. Our results collectively suggest, PI3K/AKT/mTOR, ERK/mTOR and p53/mTOR signaling pathways are involved in the regulation of SW-induced autophagy in HT22 cells, while the AMPK/mTOR signaling pathway is not involved in this regulation. Inhibition of autophagic degradation can effectively alleviate SW-induced vacuolar degeneration.


Assuntos
Autofagia , Fosfatidilinositol 3-Quinases , Swainsonina , Animais , Camundongos , Proteínas Quinases Ativadas por AMP/metabolismo , Autofagia/efeitos dos fármacos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Swainsonina/toxicidade , Serina-Treonina Quinases TOR/metabolismo , Proteína Supressora de Tumor p53/metabolismo
14.
Microb Cell Fact ; 21(1): 171, 2022 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-35999567

RESUMO

BACKGROUND: Salmonella is a common foodborne pathogen, which can cause intestinal diseases. In the last decades, the overuse of antibiotics has led to a pandemic of drug-resistant bacterial infections. To tackle the burden of antimicrobial resistant pathogens, it is necessary to develop new antimicrobial drugs with novel modes of action. However, the research and development of antibiotics has encountered bottlenecks, scientific hurdles in the development process, as well as safety and cost challenges. Phages and phage endolysins are promising antibacterial agents that can be used as an alternative to antibiotics. In this context, the expression of endolysin derived from different phages through microbial cells as a chassis seems to be an attractive strategy. RESULTS: In this study, a new endolysin from the Salmonella phage XFII-1, named XFII, was screened and obtained. The endolysin yield exceeded 100 mg/mL by heterologous expression from E. coli BL21 and short induction. The endolysin XFII exhibited high bactericidal activity at a concentration of 0.5 µg/mL and reduced the OD600 nm of EDTA-pretreated E. coli JM109 from 0.8 to 0.2 within 5 min. XFII exhibited good thermo-resistance, as it was very stable at different temperatures from 20 to 80℃. Its bactericidal activity could keep constant at 4 °C for 175 days. In addition, the endolysin was able to exert lytic activity in eutrophic conditions, including LB medium and rabbit serum, and the lytic activity was even increased by 13.8% in 10% serum matrices. XFII also showed bactericidal activity against many Gram-negative bacteria, including Salmonella, E. coli, Acinetobacter baumannii, and Klebsiella pneumoniae. Surprisingly, the combination of endolysin XFII and chitosan showed a strong synergy in lysing E. coli and Salmonella without EDTA-pretreatment, and the OD600 nm of E. coli decreased from 0.88 to 0.58 within 10 min. CONCLUSIONS: The novel globular endolysin XFII was screened and successfully expressed in E. coli BL21. Endolysin XFII exhibits a broad lysis spectrum, a rapid and strong bactericidal activity, good stability at high temperatures and under eutrophic conditions. Combined with chitosan, XFII could spontaneously lyse Gram-negative bacteria without pretreatment. This work presented the first characterization of combining endolysin and chitosan in spontaneously lysing Gram-negative bacteria in vitro.


Assuntos
Bacteriófagos , Quitosana , Animais , Antibacterianos/farmacologia , Bacteriófagos/genética , Quitosana/farmacologia , Ácido Edético , Endopeptidases/genética , Endopeptidases/metabolismo , Endopeptidases/farmacologia , Escherichia coli/genética , Escherichia coli/metabolismo , Bactérias Gram-Negativas/metabolismo , Coelhos , Salmonella/genética , Salmonella/metabolismo
15.
Biotechnol Bioeng ; 119(10): 2731-2742, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35859248

RESUMO

With the increasingly serious drug resistance of Acinetobacter baumannii, there is an increasingly urgent need for new antibacterial drugs. Phage lysin PlyAB1 has a bactericidal effect on drug-resistant A. baumannii, which has the potential to replace antibiotics to fight infection caused by A. baumannii. However, its application is limited by its thermal stability and lytic activity. To solve these problems, molecular dynamics (MD) simulations combined with Hotspot wizard 3.0 were used to identify key residue sites affecting thermal stability, and evolutionary analysis combined with multiple sequence alignment was used to identify key residue sites affecting lytic activity. Four single-point variants with significantly increased thermal stability and four single-point variants with significantly lytic activity were obtained, respectively. Furthermore, by superimposing mutations, we obtained three double-point variants, G100Q/K69R, G100R/K69R, and G100K/K69R, with significantly improved thermal stability and improved lytic activity. At 45°C, the lytic activity and half-life of the optimal variant G100Q/K69R were 1.51- and 24-fold higher than those of the wild PlyAB1, respectively. These results deepen our understanding of the structure and function of phage lysin and contribute to the application of phage lysin in antibiotic substitution.


Assuntos
Acinetobacter baumannii , Bacteriófagos , Antibacterianos/farmacologia , Bacteriófagos/genética , Mucoproteínas/farmacologia
16.
Comput Intell Neurosci ; 2022: 3968607, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35694604

RESUMO

This paper aims to explore the seismic mechanical properties of newly developed fabricated aerated lightweight concrete (ALC) wall panels to clarify the interaction mechanism between wall panels and structures. It first introduces the lightweight deep learning object detection algorithm and constructs a network model with faster operation speed based on the convolutional neural network. Secondly, combined with the deep learning object detection algorithm, the quasi-static loading system is adopted to conduct the repeated loading test on two fabricated ALC wall panels. Finally, the hysteresis load-displacement curve of each test is recorded. The experimental results show that the proposed deep learning algorithm greatly improves the operation speed and compresses the model size without reducing the accuracy. The lightweight deep learning algorithm is applied to the study of the slip performance of the wall plate. The pretightening force of the connecting screw characterizes the slip performance between the wall plate and the structural beam, thereby affecting the deformation response of the wall plate when the interstory displacement increases. The hysteresis curve of the ALC wall panel has obvious squeezing effect, indicating that the slip of the connector can unload part of the external load and delay the damage of the wall panel. The skeleton curve suggests that the fabricated windowless ALC wall panel has higher positive and negative initial stiffness and bearing capacity than the fabricated windowed wall panel. However, the degradation analysis of the stiffness curve reveals that the lateral stiffness deviation of the fabricated windowless ALC wall panel is more obvious. It confirms that the proposed connection method based on the lightweight deep learning model can improve the seismic performance of ALC wall panels and provide reference for the structural analysis of embedding fabricated ALC wall panels. This work shows the important practical value for exploring the application effect of embedded ALC wall panels.

17.
BMC Plant Biol ; 22(1): 118, 2022 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-35291945

RESUMO

BACKGROUND: The Artemisia species are widely distributed around the world, and have found important usage in traditional medicinal practice. This study was designed to investigate the metabolites of Tibetan Artemisia species and understand the metabolic pathways. METHODS: The metabolites from three Artemisia species in Tibet, were analyzed using LC-MS/MS. The differential metabolites were classified and analyzed by principal component analysis (PCA), partial least squares analysis and hierarchical clustering. KEGG Pathway enrichment analysis was used to identify the key metabolic pathways involved in the differential metabolites of three Artemisia species. RESULT: The metabolites of three Artemisia species were analyzed. Under the positive ion mode in LC-MS/MS, 262 distinct metabolites were differentially detected from Artemisia sieversiana and Artemisia annua, 312 differential metabolites were detected from Artemisia wellbyi and Artemisia sieversiana, 306 differential metabolites were screened from Artemisia wellbyi and Artemisia annua. With the negative ion mode, 106 differential metabolites were identified from Artemisia sieversiana and Artemisia annua, 131 differential metabolites were identified from Artemisia wellbyi and Artemisia sieversiana,133 differential metabolites were differentially detected from Artemisia wellbyi and Artemisia annua. The selected differential metabolites were mainly organic acids and their derivatives, ketones, phenols, alcohols and coumarins. Among these natural compounds, artemisinin, has the highest relative content in Artemisia annua. CONCLUSIONS: This is the first reported attempt to comparatively determine the types of the metabolites of the three widely distributed Artemisia species in Tibet. The information should help medicinal research and facilitate comprehensive development and utilization of Artemisia species in Tibet.


Assuntos
Artemisia annua , Espectrometria de Massas em Tandem , Artemisia annua/genética , China , Cromatografia Líquida , Metabolômica , Tibet
18.
Front Plant Sci ; 13: 1106219, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36684762

RESUMO

Sugar transporter proteins (STPs) play vital roles in sugar transport and allocation of carbon sources in plants. However, the evolutionary dynamics of this important gene family and their functions are still largely unknown in citrus, which is the largest fruit crop in the world. In this study, fourteen non-redundant CgSTP family members were identified in pummelo (Citrus grandis). A comprehensive analysis based on the biochemical characteristics, the chromosomal location, the exon-intron structures and the evolutionary relationships demonstrated the conservation and the divergence of CgSTPs. Moreover, CgSTP4, 11, 13, 14 were proofed to be localized in plasma membrane and have glucose transport activity in yeast. The hexose content were significantly increased with the transient overexpression of CgSTP11 and CgSTP14. In addition, antisense repression of CgSTP4 induced the shorter pollen tube length in vitro, implying the potential role of CgSTP4 in pummelo pollen tube growth. Taken together, this work explored a framework for understanding the physiological role of CgSTPs and laid a foundation for future functional studies of these members in citrus species.

19.
Food Chem ; 346: 128860, 2021 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-33385915

RESUMO

Molasses is a major by-product of sugar industry and contains 40-60% (w/w) of sugars. The world's annual yield of molasses reaches 55 million tons. Traditionally, molasses is simply discharged or applied to feed production. Additionally, some low-cost and environmentally friendly bioprocesses have been established for microbial production of value-added bioproducts from molasses. Over the last decade and more, increasing numbers of biofuels, polysaccharides, oligosaccharides, organic acids, and enzymes have been produced from the molasses through microbial conversion that possess an array of important applications in the industries of food, energy, and pharmaceutical. For better application, it is necessary to comprehensively understand the research status of bioconversion of molasses that has not been elaborated in detail so far. In this review, these value-added bioproducts and enzymes obtained through bioconversion of molasses, their potential applications in food and other industries, as well as the future research focus were generalized and discussed.


Assuntos
Enzimas/metabolismo , Indústria Alimentícia , Melaço , Açúcares/metabolismo , Biocombustíveis , Fermentação , Oligossacarídeos/biossíntese
20.
Sensors (Basel) ; 20(22)2020 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-33202959

RESUMO

Image matching forms an essential means of data association for computer vision, photogrammetry and remote sensing. The quality of image matching is heavily dependent on image details and naturalness. However, complex illuminations, denoting extreme and changing illuminations, are inevitable in real scenarios, and seriously deteriorate image matching performance due to their significant influence on the image naturalness and details. In this paper, a spatial-frequency domain associated image-optimization method, comprising two main models, is specially designed for improving image matching with complex illuminations. First, an adaptive luminance equalization is implemented in the spatial domain to reduce radiometric variations, instead of removing all illumination components. Second, a frequency domain analysis-based feature-enhancement model is proposed to enhance image features while preserving image naturalness and restraining over-enhancement. The proposed method associates the advantages of the spatial and frequency domain analyses to complete illumination equalization, feature enhancement and naturalness preservation, and thus acquiring the optimized images that are robust to the complex illuminations. More importantly, our method is generic and can be embedded in most image-matching schemes to improve image matching. The proposed method was evaluated on two different datasets and compared with four other state-of-the-art methods. The experimental results indicate that the proposed method outperforms other methods under complex illuminations, in both matching performances and practical applications such as structure from motion and multi-view stereo.

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