AIM: To evaluate the effect of oral Escherichia coli (E. coli) Nissle application on the outcome of intestinal-borne dermatoses.METHODS: In a randomized, controlled, non-blinded prospective clinical trial 82 patients ...AIM: To evaluate the effect of oral Escherichia coli (E. coli) Nissle application on the outcome of intestinal-borne dermatoses.METHODS: In a randomized, controlled, non-blinded prospective clinical trial 82 patients with intestinal-borne facial dermatoses characterized by an erythematous papular-pustular rash were screened. At the initiation visit 37 patients entered the experimental arm and 20 patients constituted the control arm. All 57 patients were treated with a vegetarian diet and conventional topical therapy of the dermatoses with ointments containing tetracycline, steroids and retinoids. In the experimental arm patients received a one month therapy with oral E. coli Nissle at a maintenance dose of 2 capsules daily. The experimental group was compared to a non-treatment group only receiving the diet and topical therapy. The primary outcome parameter was improvement of the dermatoses, secondary parameters included life quality and adverse events. In addition the immunological reaction profile (IgA, interleucin-8 and interferon-α) was determined. Furthermore the changes of stool consistency and the microbiota composition over the time of intervention were recorded.RESULTS: Eighty-nine percent of the patients with acne, papular-pustular rosacea and seborrhoic dermatitis responded to E. coli Nissle therapy with significant amelioration or complete recovery in contrast to 56% in the control arm (P < 0.01). Accordingly, in the E. coli Nissle treated patients life quality improved significantly (P < 0.01), and adverse events were not recorded. The clinical improvement was associated with a significant increase of IgA levels to normal values in serum as well as suppression of the proinflammatory cytokine IL-8 (P < 0.01 for both parameters). In the E. coli Nissle treated group a shift towards a protective microbiota with predominance of bifidobacteria and lactobacteria (> 10<sup>7</sup> CFU/g stool) was observed in 79% and 63% of the patients, respectively (P < 0.01), compared to no change in the control group without E. coli Nissle. Moreover, the detection rate of a pathogenic flora dropped from 73% to 14 % of the patients in the experimental arm (P < 0.01) with no significant change in the control arm (accounting 80% before and 70% after the observation period, P > 0.05). Accordingly, stool consistency, color and smell normalized in the E. coli Nissle treated patients.CONCLUSION: E. coli Nissle protects the mucus barrier by overgrowth of a favorable gut microbiota with less immunoreactive potential which finally leads to clinical improvement of intestinal borne dermatoses.展开更多
AIM： To evaluate bacteriocinogeny in short-term highdose indomethacin administration with or without probiotic Escherichia coli Nissle 1917 （EcN） in experimental pigs.METHODS： Twenty-four pigs entered the study： ...AIM： To evaluate bacteriocinogeny in short-term highdose indomethacin administration with or without probiotic Escherichia coli Nissle 1917 （EcN） in experimental pigs.METHODS： Twenty-four pigs entered the study： Group A （controls）, Group B （probiotics alone）, Group C （indomethacin alone） and Group D （probiotics and indomethacin）. EcN （3.5 × 10^10 bacteria/d for 14 d） and/or indomethacin （15 mg/kg per day for 10 d） were administrated orally. Anal smears before and smears from the small and large intestine were taken from all animals. Bacteriocin production was determined with 6 different indicator strains; all strains were polymerase chain reaction tested for the presence of 29 individual bacteriocinencoding determinants. RESULTS： The general microbiota profile was rather uniform in all animals but there was a broad diversity in coliform bacteria （parallel genotypes A, B1, B2 and D found）. In total, 637 bacterial strains were tested, mostly Escherichia coli （E. coli. There was a higher incidence of non-E：, coli strains among samples taken from the jejunum and ileum compared to that of the colon and rectum indicating predominance of E. coil strains in the large intestine. Bacteriocinogeny was found in 24/77 （31%） before and in 155/560 （28%） isolated bacteria at the end of the study. Altogether, 13 individual bacteriocin types （out of 29 tested） were identified among investigated strains. Incidence of four E. coli genotypes was equally distributed in all groups of E. coil strains, with majority of genotype A （ranging from 81% to 88%）. The following types of bacteriocins were most commonly revealed： colicins Ia/Ib （44%）, microcin V （18%）, colicin E1 （16%） and microcin H47 （6%）. There was a difference in bacteriocinogeny between control group A （52/149, 35%） and groups with treatment at the end of the study： B： 31/122 （25%, P = 0.120）; C： 43/155 （28%, P = 0.222）; D： 29/134 （22%, P = 0.020）. There was a significantly lower prevalence of colicin Ib, microcins H47 and V （probiotics group, P 〈 0.001）, colicin E1 and microcin H47 （indomethacin group, P 〈 0.001） and microcins H47 and V （probiotics and indomethacin group, P = 0.025） compared to controis. Escherichia fergusonil （E. fergusoniO was identi-fled in 6 animals （6/11 isolates from the rectum）. One strain was non-colicinogenic, while all other strains of E. fergusonii solely produced colicin El. All animals started and remained methanogenic despite the fact that EcN is a substantial hydrogen producer. There was an increase in breath methane （after the treatment） in 5/6 pigs from the indomethacin group （C）. CONCLUSION： EcN did not exert long-term liveabilib/ in the porcine intestine, All experimental pigs remained methanogenic, Indomethacin and EcN administered together might produce the worst impact on bacteriocinogeny.展开更多
Escherichia coli is the most well-studied model prokaryote and has become an indispensable host for the biotech-nological production of proteins and biochemicals.In particular,the probiotic status of one E.coli strain...Escherichia coli is the most well-studied model prokaryote and has become an indispensable host for the biotech-nological production of proteins and biochemicals.In particular,the probiotic status of one E.coli strain,E.coli Nissle 1917(EcN)has helped it become a new favorite amongst synthetic biologists.To broaden its potential applications,here we assemble a comparative study on the genomes,transcriptomes,and metabolic properties of E.coli strains EcN,BL21(DE3),and MG1655.Comparative genomics data suggests that EcN possesses 1404 unique CDSs.In particular,EcN has additional iron transport systems which endow EcN with a higher tolerance to iron scarcity when compared to two other E.coli strains.EcN transcriptome data demonstrates that E.coli strains EcN,BL21(DE3),and MG1655 all have comparable activities of the central metabolic pathway,however only EcN inherits the arginine deiminase pathway.Additionally,we found that EcN displayed a lower expres-sion of ribosomal proteins compared to BL21(DE3)and MG1655.This comparative study on E.coli strains EcN,BL21(DE3),and MG1655 aims to provide a reference for further engineering EcN as a biotechnological tool.展开更多
质粒消除旨在获得无质粒菌株,是对益生菌进行遗传改造所需的一项重要技术。本试验建立在质粒不相容性的基础上,引入自杀性载体pRE112作为辅助工具,使用分子生物学方法构建重组自杀质粒pMUT1-pRE112和pMUT2-pRE112,将其分别导入携带2个...质粒消除旨在获得无质粒菌株,是对益生菌进行遗传改造所需的一项重要技术。本试验建立在质粒不相容性的基础上,引入自杀性载体pRE112作为辅助工具,使用分子生物学方法构建重组自杀质粒pMUT1-pRE112和pMUT2-pRE112,将其分别导入携带2个大隐秘质粒pMUT1和pMUT2的益生菌大肠杆菌Nissle 1917(Escherichia coli Nissle 1917,EcN),利用重组自杀质粒去除EcN内原有隐秘质粒并在含有10%蔗糖的LB培养基上实施自杀质粒自身消除。结果试验成功获得EcN无质粒克隆菌株(Escherichia coli Nissle 1917cured of its two cryptic plasmids pMUT1and pMUT2,EcNc),为优质益生菌EcN更好的潜在应用研究奠定基础。展开更多
Colitis is a common disease of the colon that is very difficult to treat.Probiotic bacteria could be an effective treatment.The probiotic Escherichia coli Nissle 1917(EcN)was engineered to synthesize the ketone body(R...Colitis is a common disease of the colon that is very difficult to treat.Probiotic bacteria could be an effective treatment.The probiotic Escherichia coli Nissle 1917(EcN)was engineered to synthesize the ketone body(R)-3-hydroxybutyrate(3HB)for sustainable production in the gut lumen of mice suffering from colitis.Components of heterologous 3HB synthesis routes were constructed,expressed,optimized,and inserted into the EcN genome,combined with deletions in competitive branch pathways.The genome-engineered EcN produced the highest 3HB level of 0.6 g/L under microaerobic conditions.The live therapeutic was found to colonize the mouse gastrointestinal tract over 14 days,elevating gut 3HB and short-chain-length fatty acid(SCFA)levels 8.7-and 3.1-fold compared to those of wild-type EcN,respectively.The sustainable presence of 3HB in mouse guts promoted the growth of probiotic bacteria,especially Akkermansia spp.,to over 31%from the initial 2%of all the microbiome.As a result,the engineered EcN termed EcNL4 ameliorated colitis induced via dextran sulfate sodium(DSS)in mice.Compared to wild-type EcN or oral administration of 3HB,oral EcNL4 uptake demonstrated better effects on mouse weights,colon lengths,occult blood levels,gut tissue myeloperoxidase activity and proinflammatory cytokine concentrations.Thus,a promising live bacterium was developed to improve colonic microenvironments and further treat colitis.This proof-of-concept design can be employed to treat other diseases of the colon.展开更多
文摘AIM: To evaluate the effect of oral Escherichia coli (E. coli) Nissle application on the outcome of intestinal-borne dermatoses.METHODS: In a randomized, controlled, non-blinded prospective clinical trial 82 patients with intestinal-borne facial dermatoses characterized by an erythematous papular-pustular rash were screened. At the initiation visit 37 patients entered the experimental arm and 20 patients constituted the control arm. All 57 patients were treated with a vegetarian diet and conventional topical therapy of the dermatoses with ointments containing tetracycline, steroids and retinoids. In the experimental arm patients received a one month therapy with oral E. coli Nissle at a maintenance dose of 2 capsules daily. The experimental group was compared to a non-treatment group only receiving the diet and topical therapy. The primary outcome parameter was improvement of the dermatoses, secondary parameters included life quality and adverse events. In addition the immunological reaction profile (IgA, interleucin-8 and interferon-α) was determined. Furthermore the changes of stool consistency and the microbiota composition over the time of intervention were recorded.RESULTS: Eighty-nine percent of the patients with acne, papular-pustular rosacea and seborrhoic dermatitis responded to E. coli Nissle therapy with significant amelioration or complete recovery in contrast to 56% in the control arm (P < 0.01). Accordingly, in the E. coli Nissle treated patients life quality improved significantly (P < 0.01), and adverse events were not recorded. The clinical improvement was associated with a significant increase of IgA levels to normal values in serum as well as suppression of the proinflammatory cytokine IL-8 (P < 0.01 for both parameters). In the E. coli Nissle treated group a shift towards a protective microbiota with predominance of bifidobacteria and lactobacteria (> 10<sup>7</sup> CFU/g stool) was observed in 79% and 63% of the patients, respectively (P < 0.01), compared to no change in the control group without E. coli Nissle. Moreover, the detection rate of a pathogenic flora dropped from 73% to 14 % of the patients in the experimental arm (P < 0.01) with no significant change in the control arm (accounting 80% before and 70% after the observation period, P > 0.05). Accordingly, stool consistency, color and smell normalized in the E. coli Nissle treated patients.CONCLUSION: E. coli Nissle protects the mucus barrier by overgrowth of a favorable gut microbiota with less immunoreactive potential which finally leads to clinical improvement of intestinal borne dermatoses.
基金Supported by Research project MZO 00179906 from the Ministry of Health of the Czech RepublicSupported by institutional support from the Czech Republic(MSM0021622415)Supported by research grants GAR 305/080535 and NS9665-4/2008(Ministry of Health of the Czech Republic)
文摘AIM： To evaluate bacteriocinogeny in short-term highdose indomethacin administration with or without probiotic Escherichia coli Nissle 1917 （EcN） in experimental pigs.METHODS： Twenty-four pigs entered the study： Group A （controls）, Group B （probiotics alone）, Group C （indomethacin alone） and Group D （probiotics and indomethacin）. EcN （3.5 × 10^10 bacteria/d for 14 d） and/or indomethacin （15 mg/kg per day for 10 d） were administrated orally. Anal smears before and smears from the small and large intestine were taken from all animals. Bacteriocin production was determined with 6 different indicator strains; all strains were polymerase chain reaction tested for the presence of 29 individual bacteriocinencoding determinants. RESULTS： The general microbiota profile was rather uniform in all animals but there was a broad diversity in coliform bacteria （parallel genotypes A, B1, B2 and D found）. In total, 637 bacterial strains were tested, mostly Escherichia coli （E. coli. There was a higher incidence of non-E：, coli strains among samples taken from the jejunum and ileum compared to that of the colon and rectum indicating predominance of E. coil strains in the large intestine. Bacteriocinogeny was found in 24/77 （31%） before and in 155/560 （28%） isolated bacteria at the end of the study. Altogether, 13 individual bacteriocin types （out of 29 tested） were identified among investigated strains. Incidence of four E. coli genotypes was equally distributed in all groups of E. coil strains, with majority of genotype A （ranging from 81% to 88%）. The following types of bacteriocins were most commonly revealed： colicins Ia/Ib （44%）, microcin V （18%）, colicin E1 （16%） and microcin H47 （6%）. There was a difference in bacteriocinogeny between control group A （52/149, 35%） and groups with treatment at the end of the study： B： 31/122 （25%, P = 0.120）; C： 43/155 （28%, P = 0.222）; D： 29/134 （22%, P = 0.020）. There was a significantly lower prevalence of colicin Ib, microcins H47 and V （probiotics group, P 〈 0.001）, colicin E1 and microcin H47 （indomethacin group, P 〈 0.001） and microcins H47 and V （probiotics and indomethacin group, P = 0.025） compared to controis. Escherichia fergusonil （E. fergusoniO was identi-fled in 6 animals （6/11 isolates from the rectum）. One strain was non-colicinogenic, while all other strains of E. fergusonii solely produced colicin El. All animals started and remained methanogenic despite the fact that EcN is a substantial hydrogen producer. There was an increase in breath methane （after the treatment） in 5/6 pigs from the indomethacin group （C）. CONCLUSION： EcN did not exert long-term liveabilib/ in the porcine intestine, All experimental pigs remained methanogenic, Indomethacin and EcN administered together might produce the worst impact on bacteriocinogeny.
基金supported by the National Key Re-search and Development Program of China(2021YFC2100800)the Jiangsu Province Natural Science Fund for Distinguished Young Schol-ars(BK20200025)+1 种基金a grant from the Key Technologies R&D Program of Jiangsu Province(BE2019630)the National First-class Discipline Program of Light Industry Technology and Engineering(LITE2018-16).
文摘Escherichia coli is the most well-studied model prokaryote and has become an indispensable host for the biotech-nological production of proteins and biochemicals.In particular,the probiotic status of one E.coli strain,E.coli Nissle 1917(EcN)has helped it become a new favorite amongst synthetic biologists.To broaden its potential applications,here we assemble a comparative study on the genomes,transcriptomes,and metabolic properties of E.coli strains EcN,BL21(DE3),and MG1655.Comparative genomics data suggests that EcN possesses 1404 unique CDSs.In particular,EcN has additional iron transport systems which endow EcN with a higher tolerance to iron scarcity when compared to two other E.coli strains.EcN transcriptome data demonstrates that E.coli strains EcN,BL21(DE3),and MG1655 all have comparable activities of the central metabolic pathway,however only EcN inherits the arginine deiminase pathway.Additionally,we found that EcN displayed a lower expres-sion of ribosomal proteins compared to BL21(DE3)and MG1655.This comparative study on E.coli strains EcN,BL21(DE3),and MG1655 aims to provide a reference for further engineering EcN as a biotechnological tool.
文摘质粒消除旨在获得无质粒菌株,是对益生菌进行遗传改造所需的一项重要技术。本试验建立在质粒不相容性的基础上,引入自杀性载体pRE112作为辅助工具,使用分子生物学方法构建重组自杀质粒pMUT1-pRE112和pMUT2-pRE112,将其分别导入携带2个大隐秘质粒pMUT1和pMUT2的益生菌大肠杆菌Nissle 1917(Escherichia coli Nissle 1917,EcN),利用重组自杀质粒去除EcN内原有隐秘质粒并在含有10%蔗糖的LB培养基上实施自杀质粒自身消除。结果试验成功获得EcN无质粒克隆菌株(Escherichia coli Nissle 1917cured of its two cryptic plasmids pMUT1and pMUT2,EcNc),为优质益生菌EcN更好的潜在应用研究奠定基础。
基金This research was financially supported by a grant from the Chunfeng Foundation(2020Z99CFG002)of Tsinghua UniversityOther support includes the National Natural Science Foundation of China(Grant Nos.31870859,21761132013,31771886,and 31971170).
文摘Colitis is a common disease of the colon that is very difficult to treat.Probiotic bacteria could be an effective treatment.The probiotic Escherichia coli Nissle 1917(EcN)was engineered to synthesize the ketone body(R)-3-hydroxybutyrate(3HB)for sustainable production in the gut lumen of mice suffering from colitis.Components of heterologous 3HB synthesis routes were constructed,expressed,optimized,and inserted into the EcN genome,combined with deletions in competitive branch pathways.The genome-engineered EcN produced the highest 3HB level of 0.6 g/L under microaerobic conditions.The live therapeutic was found to colonize the mouse gastrointestinal tract over 14 days,elevating gut 3HB and short-chain-length fatty acid(SCFA)levels 8.7-and 3.1-fold compared to those of wild-type EcN,respectively.The sustainable presence of 3HB in mouse guts promoted the growth of probiotic bacteria,especially Akkermansia spp.,to over 31%from the initial 2%of all the microbiome.As a result,the engineered EcN termed EcNL4 ameliorated colitis induced via dextran sulfate sodium(DSS)in mice.Compared to wild-type EcN or oral administration of 3HB,oral EcNL4 uptake demonstrated better effects on mouse weights,colon lengths,occult blood levels,gut tissue myeloperoxidase activity and proinflammatory cytokine concentrations.Thus,a promising live bacterium was developed to improve colonic microenvironments and further treat colitis.This proof-of-concept design can be employed to treat other diseases of the colon.
