Frontiers in Cellular and Infection Microbiology
·
2025
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IF 4.8
Yizhuo Zhang,Luqin Lv,Su Xu,Yijian Chen
Introduction: Clostridioides difficile (C. difficile), a human pathogen that causes diarrhea and colon lesions, has garnered widespread attention. Rapid and accurate detection of bacterial virulence factors is essential for the diagnosis of C. difficile infection (CDI). To date, numerous laboratory tests have been developed; however, none fully meet the combined requirements of speed, cost-effectiveness, portability, sensitivity, and specificity. Molecular diagnostic technologies based on CRISPR-Cas systems have provided a promising solution to this challenge. Nonetheless, the limited compatibility between pre-amplification and CRISPR cleavage, coupled with the inherent selectivity of CRISPR systems for protospacer adjacent motif (PAM) sequences near the target site, poses additional constraints on the broader adoption of this approach.Methods: Here, we developed PAM-unconventional, One-step LAMP/CRISPR-Cas12b (POLC) detection platforms for the toxin-encoding genes tcdA and tcdB of C. difficile.Results: The POLC platforms operated at 60 °C, enabling result interpretation either through fluorescence intensity measurements or direct visualization under UV light. The limits of detection (LoDs) ranged from 3 to 14 copies/μL using a fluorescence reader and from 6 to 18 copies/μL via direct observation. Compared to qPCR, which typically requires over an hour, the POLC platforms reduced the detection time to approximately 40 minutes. Each reaction cost approximately USD 6.5, offering a substantial cost saving compared to qPCR-based commercial kits (over USD 10 per test). In clinical validation with 55 fecal samples, the tcdA POLC assay achieved 86.4% sensitivity and 84.8% specificity, while the tcdB POLC assay demonstrated 96.6% sensitivity and 100% specificity, using qPCR as the reference standard.Discussion: Our research presents innovative CRISPR-based one-step nucleic acid detection platforms that eliminate canonical PAM sequence requirements. These platforms exhibit high sensitivity and specificity while achieving rapid detection under simple conditions, making them promising candidates for clinical diagnostics and point-of-care testing (POCT).
研究领域:
医学诊断
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分子生物学
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产品关联
DOI
PubMed
高影响因子
MOLECULES
·
2023
·Vol. 28
·期 16
·6152
·
IF 4.6
Lijun Xie,Li Chen,Yongbo Wei,Nannan Chen,Tong Wu,Jingming Zhou,Hong Jiang,Feng Lin
Rakicidin B1 was isolated and purified from the culture broth of a marineStreptomycessp. as a potent anti-cancer agent, and lately the compound and its derivatives have firstly been found to possess anti-Clostridium difficile(CD) activity but with high cytotoxicity. Herein, following our previous discovery on anti-CD activity of Rakicidin B1, structure modification was performed at the OH position of Rakicidin B1 and a new Rakicidin B1-PEG hybrids FIMP2 was facilely designed and synthesized by conjugating the PEG2000 with the scaffolds of Rakicidin B1 via the linkage of carbamate. The cytotoxicity of the FIMP2 was first evaluated against three different cancer cell lines, including HCT-8 cells, PANC-1, and Caco-2, with IC50values at 0.519 μM, 0.815 μM, and 0.586 μM, respectively. Obviously, as compared with a positive control group treated with Rakicidin B1, the IC50value of FIMP2 increased by nearly 91-fold, 50-fold, and 67-fold, suggesting that the PEGylation strategy significantly reduced the cytotoxicity of FIMP2. Thus, this preliminary result may be beneficial to increase its safety index (SI) value due to the decreased cytotoxicity of FIMP2. In addition, this decreased cytotoxicity of FIMP2 was further confirmed based on a zebrafish screening model in vivo. Thereafter, the anti-CD activity of FIMP2 was evaluated in vivo, and its efficacy to treat CDI was found to be better than that of vancomycin. The mortality and recurrence rate of FIMP2 is not as low compared with that of vancomycin; these results demonstrated that compound FIMP2 is a new, promising anti-CD agent with significant efficacy against CD recurrence with low cytotoxicity towards bodies.Keywords:Rakicidin B1;PEGylation strategy;anti-Clostridium difficile;low cytotoxicity
研究领域:
药物化学
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微生物学
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药理学
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DOI
PubMed
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2019
·
IF 3.579
Nan Zhang,Tisong Liang,Quan Jin,Cai Shen,Yifan Zhang,Pu Jing
Antibiotic-associated diarrhea (AAD) is a common adverse effect of antibiotic treatment. The study was to evaluate effects of Chinese yam ( Dioscorea opposita Thunb.) on AAD, and changes of intestinal microbiota and short-chain fatty acids (SCFA). AAD model was established using Balb/c mice by gavage with ampicillin for 5 days, followed by a 10-day administration of low, medium, and high dosage of Chinese yam, containing allantoin (4.35 mg/g) and polysaccharides (85.51 mg/g). The results showed that Chinese yam accelerated the recovery from acute diarrhea, reverse AAD-induced body weight loss and cecal enlargement. The high-dosage Chinese yam increased probiotic Bifidobacteria and Lactobacilli by 47% and 21% and decreased potential pathogen Enterococcus and Clostridium perfringens by 8% and 27% on day 15 ( P < .05). Bacterial community analysis revealed that Chinese yam contributed to repair the ampicillin-induced intestinal microbiota disorder, enrich the abundance of Bacteroides spp. and Clostridium spp. Additionally, Chinese yam supplementation increased the production of SCFA.
研究领域:
药理学
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微生物学
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营养学
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DOI
PubMed
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
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2011
·Vol. 165
·期 2
·666-675
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IF 1.879
Chen Sheng,Liu Zhiguo,Chen Jian,Wu Jing
Escherichia coli is one of the most commonly used host strains for recombinant protein production. More and more research works on the production of recombinant protein indicate that extracellular production throughout a culture medium is more convenient and attractive compared to intracellular production. In present work, inducing temperature and isopropyl β- d -1-thiogalactopyranoside (IPTG) concentration were investigated to decrease the formation of inclusion body and increase the amount of soluble recombinant cutinase initially. Enzyme activity in the culture medium reached to 118.9 U/ml at 64 h of culture, and no inclusion body was detected in cytoplasm under the inducement condition of 0.2 mM IPTG and 30°C. In addition, it was found that a large amount of cutinase had been accumulated in periplasm since 16-h cultivation under the same inducement condition. Therefore, glycine and surfactant sodium taurodeoxycholate (TDOC) were further used to promote the leakage of recombinant cutinase from periplasm. Supplied with 100 mM glycine and 1 mM TDOC, the amount of cutinase in periplasm decreased remarkably, and the activity in the culture medium reached to 146.2 and 149.2 U/ml after 54 h of culturing, respectively.
研究领域:
生物工程
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分子生物学
✓ 已验证
DOI
高影响因子
Gut Microbes
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2025
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IF 11
Yan Liu,Zhangtie Wang,Yansong Zhang,Tian Zhao,Qingjun Zhang,Weisu Huang,Baiyi Lu
Dietary cholesterol is often found in a high-fat diet (HFD) and excessive intake is harmful to cognitive function. The gut microbiome constitutes an environmental factor influenced by diet, which regulates cognitive function via the gut-brain axis. The present study explored the role of dietary cholesterol in HFD-induced cognitive impairment and the participation of the gut microbiota and metabolites. Here, we found that dietary cholesterol promoted cognitive impairment in HFD-fed mice, which was associated with an increase in gut microbiota containing 7α-dehydroxylase, including Lachnospiraceae bacterium, Dorea sp. Clostridium sp. and elevated levels of deoxycholic acid (DCA) in the hippocampus. Upon dietary cholesterol intake, the activity of gut microbiota in mice to produce DCA is increased. Fecal microbiota transplantation confirmed that the cognitive impairment-promoting process was driven by gut microbiota. Reducing circulating bile acid levels with cholestyramine improved cognitive decline in mice, whereas hippocampal administration of DCA worsened cognitive function. Pharmacological inhibition of hippocampal apical sodium bile acid transporter reduces neuronal DCA accumulation and improves neuronal apoptosis as well as cognitive impairments in mice. Overall, this study revealed that dietary cholesterol promotes HFD-induced cognitive impairment by inducing the production of DCA through gut microbiota metabolism.
研究领域:
神经科学
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微生物学
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营养学
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代谢组学
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药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Biomass Conversion and Biorefinery
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2020
·Vol. 13
·期 2
·647-660
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IF 2.602
Nalawade Ketaki,Saikia Paharika,Behera Shuvashish,Konde Kakasaheb,Patil Sanjay
The bioprospecting of sugarcane bagasse (SCB) through alkali, acid, and hydrodynamic cavitation pretreatment methods and their combinations were evaluated based on bagasse composition, enzymatic hydrolysis, and lactic acid productivity using Bacillus coagulans NCIM 5648. From 100.0 g of SCB, L-lactic acid production of 26.16 g, 8.78 g, 14.15 g, 14.33 g, and 24.61 g in alkali, acid, sequential acid-alkali, sequential alkali-acid, and cavitation with alkali pretreatment was obtained, respectively. Considering the holistic approach from SCB to L-lactic acid, alkali pretreatment is found to be the best method with L-lactic acid titer of 68.7 g/L, the productivity of 2.86 g/L/h, and yield of 0.92 g/g which has resulted in 82.5% higher product yield from SCB as compared to alkali-acid pretreatment. Cavitation in presence of alkali evolved as the next better route with L-lactic acid titer of 62.5 g/L, the productivity of 2.60 g/L/h, and yield of 0.92 g/g. Though the highest glucose release of 89.3 g/L was achieved during enzymatic hydrolysis with sequential alkali-acid-pretreated SCB that resulted in the highest L-lactic acid titer of 71.8 g/L, the productivity of 2.99 g/L/h and fermentation yield of 0.90 g/g.
研究领域:
生物技术
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可再生能源
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发酵工程
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DOI
高影响因子
Food Bioscience
·
2025
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IF 5.9
Liping Dong,Yonggan Sun,Shanshan Zhang,Tianxiang Ouyang,Xinyu Bian,Jiaqi Jiang,Wenting Su,Shaoping Nie
Revealing strain-level diversity in the gut microbiome is critical to understanding its impact on health. Genomic and metabolomic advances enable strain-level dissection of the gut microbiome. Faecalibacterium prausnitzii , a prevalent commensal bacterium in the human gut, is a promising candidate for next-generation probiotics. However, the study on genomic and metabolomic diversity of F. prausnitzii strains remains insufficient. In this study, 12 F. prausnitzii strains were isolated and whole genome sequencing was conducted. Results showed that these strains were classified into three phylogenetic clades (clade B, C, and D). Additionally, strain FP3003 displayed typical characteristics, marked by significant acetate consumption and butyrate production. Further integration of non-targeted metabolomics analysis and an in vitro intestinal epithelial cell model confirmed the established intestinal protective effects of F. prausnitzii and butyrate. Moreover, we identified four novel beneficial F. prausnitzii -derived metabolites, including indolelactic acid (ILA), hydroxyphenyllactic acid, hydroxyisocaproic acid, and butoxyacetic acid, among which ILA upregulated Occludin and Ecadherin expression in vitro . In summary, our results systematically elucidated the strain-level insights into the metabolic diversity and functional heterogeneity of F. prausnitzii . This study expanded the current knowledge on F. prausnitzii and established a key research paradigm for investigating the functional potential of next-generation probiotics.
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产品关联
DOI
高影响因子
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
·
2024
·Vol. 91
·期 1
·
IF 3.9
Ji Zeng,Hao Wang,Yuxi Xu,Jianying Han,Yannan Li,Shu'an Wen,Changbu Wu,Dani Li,Zheng Liu,Xiaokang Zhang,Guo-Bao Tian,Min Dong
Clostridioides difficile is an obligate anaerobic, Gram-positive bacterium that produces toxins. Despite technological progress, conducting gene expression analysis of C. difficile under different conditions continues to be labor-intensive. Therefore, there is a demand for simplified tools to investigate the transcriptional and translational regulation of C. difficile. The cell-free gene expression (CFE) system has demonstrated utility in various applications, including prototyping, protein production, and in vitro screening. In this study, we developed a C. difficile CFE system capable of in vitro transcription and translation (TX-TL) in the presence of oxygen. Through optimization of cell extract preparation and reaction systems, we increased the protein yield significantly. Furthermore, our observations indicated that this system exhibited higher protein yield using linear DNA templates than circular plasmids for in vitro expression. The prototyping capability of the C. difficile CFE system was assessed using a series of synthetic Clostridium promoters, demonstrating a good correlation between in vivo and in vitro expression. Additionally, we tested the expression of tcdB and tcdR from clinically relevant C. difficile strains using the CFE system, confirming higher toxin expression of the hypervirulent strain R20291. We believe that the CFE system can not only serve as a platform for in vitro protein synthesis and genetic part prototyping but also has the potential to be a simplified model for studying metabolic regulations in Clostridioides difficile.
研究领域:
微生物学
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分子生物学
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生物技术
✓ 已验证
DOI
高影响因子
Aquaculture Reports
·
2023
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IF 3.7
Yi Du,Long Cheng,Jianhua Zhao,Clement R. de Cruz,Hong Xu,Liansheng Wang,Qiyou Xu
This study investigated the effects of Clostridium butyricum (CB) and sodium butyrate (SB) on the growth performance, intestinal immune, and gut microbiota of mirror carp ( Cyprinus carpio ) fed with a high level of soybean meal. Three hundred sixty healthy carp fingerlings (∼3.74 g) were divided into eight dietary treatments with four replicates and ten fish per replicate. Which include basal diet (Control group, C0); C0 supplemented with three levels of CB, 3 × 10 7 cfu/kg (C1), 3 × 10 8 cfu/kg (C2), 3 × 10 9 cfu/kg (C3); three levels of SB, 500 mg/kg (S1), 1000 mg/kg (S2), 2 000 mg/kg (S3); and 3 × 10 8 cfu/kg CB+ 1 000 mg/kg SB (CS), respectively. In 8-week trial, the survival rate of all groups was 100%. Compared with C0, the Weight Gain, Feed Conversion Ratio, Protein Efficiency Ratio were not significantly different in all groups ( P > 0.05). Crude protein and ether extract contents were increased significantly ( P < 0.05) when supplemented with different levels of CB and SB, and ash content increased significantly in the CB groups ( P < 0.05). Serum globulin increased and glucose decreased significantly ( P < 0.05) in the CB and SB groups. The intestinal T-SOD and glutathione activity in the foregut, midgut, and hindgut were increased ( P < 0.05), and villus height of foregut and midgut were higher in the CB and SB ( P < 0.05). Protease activities in the foregut and midgut were significantly increased ( P < 0.05) in the C2, C3, S2, and S3. The lipase activity of the C2 and C3 in foregut, midgut, and hindgut; S2, S3, and CS in the foregut; S3 in midgut were significantly increased ( P < 0.05). The gut microbiota was modulated at the phylum and genera level by CB and SB administration. In summary, our results indicated that CB and SB could promote the whole body CP and EE content, intestine immunity, digestive enzyme activity, and intestinal morphology, and modulated gut flora in carp.
研究领域:
水产养殖
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营养学
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微生物学
·
渔业科学
✓ 已验证
产品关联
DOI
高影响因子
Frontiers in Veterinary Science
·
2022
·
IF 3.471
Lu Yixing,Yang Liuye,Zhang Wanying,Li Jie,Peng Xianfeng,Qin Zonghua,Zeng Zhenling,Zeng Dongping
This study aimed to evaluate the antibacterial activity of isopropoxy benzene guanidine (IBG) against C. perfringens based on pharmacokinetics/pharmacodynamics (PK/PD) modeling in broilers. The PK parameters of IBG in the plasma and ileal content of C. perfringens-infected broilers following oral administration at 2, 30, and 60 mg/kg body weight were investigated. in vivo PD studies were conducted over oral administration ranging from 2 to 60 mg/kg and repeated every 12 h for 3 days. The inhibitory Imax model was used for PK/PD modeling. Results showed that the MIC of IBG against C. perfringens was 0.5–32 mg/L. After oral administration of IBG, the peak concentration (Cmax), maximum concentration time (Tmax), and area under the concentration-time curve (AUC) in ileal content of broilers were 10.97–1,036.64 mg/L, 2.39–4.27 h, and 38.31–4,266.77 mg·h/L, respectively. After integrating the PK and PD data, the AUC0 − 24h/MIC ratios needed for the bacteriostasis, bactericidal activity, and bacterial eradication were 4.00, 240.74, and 476.98 h, respectively. For dosage calculation, a dosage regimen of 12.98 mg/kg repeated every 12 h for 3 days was be therapeutically effective in broilers against C. perfringens with MIC ≤ 2 mg/L. In addition, IBG showed potent activity against C. perfringens, which may be responsible for cell membrane destruction. These results can facilitate the evaluation of the use of IBG in the treatment of intestinal diseases in broilers caused by C. perfringens.
研究领域:
兽医学
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微生物学
·
药理学
✓ 已验证
DOI
PubMed
高影响因子
Food Bioscience
·
2021
·
IF 4.24
Yujin Kim,Sejeong Kim,Heeyoung Lee,Nam-Su Oh,Min Suk Rhee,Yohan Yoon
Clostridium perfringens is an important bacterium among commensal bacteria in the gastrointestinal tract , which is known to possess potential pathogenicity during its inhabitation. In concordance with the concerns raised by its pathogenicity, this study aimed to elucidate if FMRP (fermented Maillard reaction products), the novel milk-derived material, can reduce the effect caused by the overabundance of C. perfringens in the gut. As a result, FMRP significantly reduced mRNA expression levels in the inflammatory cytokines (interleukin-10, interleukin-11, and interferon-γ) (p<0.05), and tumorigenic potential by activating the cell proliferation-suppressing factors (β-catenin, adenomatous polyposis coli , and E-cadherin) (p<0.05) and deactivation of the cell proliferation-inducing factor (cyclooxygenase-2) (p<0.05) in the colon. Moreover, the tight junction proteins (zonula occludens-1 and claudin-3) were recovered (p<0.05), and the antioxidant enzymes (catalase, superoxide dismutase, and glutathione peroxidase) were decreased by FMRP (p<0.05). However, biochemical analyses showed no significant difference. These results indicate that C. perfringens might cause inflammation, oxidative stress , disruption of tight junctions and abnormal cell proliferation to the colon, but FMRP may reduce inflammation and generation of reactive oxygen, and recover tight junction in the colon.
研究领域:
微生物学
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消化系统疾病
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营养学
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产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF HYDROGEN ENERGY
·
2024
·
IF 8.1
Guoqing Liao,Chao Wang,Ao Xia,Jingmiao Zhang,Yun Huang,Xianqing Zhu,Xun Zhu,Qiang Liao
Dark fermentation is an effective means to convert organic waste to hydrogen for bioenergy utilization but is limited by non-hydrogen-producing pathways. Emerging electro-fermentation technology promises to improve it by modulating dominant metabolic pathways. This study investigated the impact of electro-fermentation performance with voltages and substrate concentrations. The results revealed that the specific hydrogen yields at 0.3–0.6 V voltage were increased by 31.5%–91.3% over the control, and substrate degradation efficiency were also improved by 13.3%–20.7%. When the voltage was applied, the abundance of the hydrogen-producing bacteria Clostridium species increased 2.6–8.1 times more than the conventional. Meanwhile, an increase of 74.6%–311.5% in electron transfer system activity was quantified with nearly doubling the thickness of the biofilm. High soluble metabolite acetate/butyrate expression provided evidence that electro-fermentation promotes both favorable hydrogen production pathways. This study provides new insights into the application of electro-fermentation as a strategy for improving sustainable hydrogen production.
研究领域:
环境工程
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生物技术
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微生物学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）