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1.
Front Bioeng Biotechnol ; 12: 1373130, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38572363

RESUMEN

Ureteral stenting is a common clinical procedure for the treatment of upper urinary tract disorders, including conditions such as urinary tract infections, tumors, stones, and inflammation. Maintaining normal renal function by preventing and treating ureteral obstruction is the primary goal of this procedure. However, the use of ureteral stents is associated with adverse effects, including surface crusting, bacterial adhesion, and lower urinary tract symptoms (LUTS) after implantation. Recognizing the need to reduce the complications associated with permanent ureteral stent placement, there is a growing interest among both physicians and patients in the use of biodegradable ureteral stents (BUS). The evolution of stent materials and the exploration of different stent coatings have given these devices different roles tailored to different clinical needs, including anticolithic, antibacterial, antitumor, antinociceptive, and others. This review examines recent advances in BUS within the last 5 years, providing an in-depth analysis of their characteristics and performance. In addition, we present prospective insights into the future applications of BUS in clinical settings.

2.
Asian J Androl ; 25(3): 366-374, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-35915542

RESUMEN

Studies have investigated the effects of androgen deprivation therapy (ADT) use on the incidence and clinical outcomes of coronavirus disease 2019 (COVID-19); however, the results have been inconsistent. We searched the PubMed, Medline, Cochrane, Scopus, and Web of Science databases from inception to March 2022; 13 studies covering 84 003 prostate cancer (PCa) patients with or without ADT met the eligibility criteria and were included in the meta-analysis. We calculated the pooled risk ratios (RRs) with 95% confidence intervals (CIs) to explore the association between ADT use and the infection risk of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and severity of COVID-19. After synthesizing the evidence, the pooled RR in the SARS-CoV-2 positive group was equal to 1.17, and the SARS-CoV-2 positive risk in PCa patients using ADT was not significantly different from that in those not using ADT (P = 0.544). Moreover, no significant results concerning the beneficial effect of ADT on the rate of intensive care unit admission (RR = 1.04, P = 0.872) or death risk (RR = 1.23, P = 0.53) were found. However, PCa patients with a history of ADT use had a markedly higher COVID-19 hospitalization rate (RR = 1.31, P = 0.015) than those with no history of ADT use. These findings indicate that ADT use by PCa patients is associated with a high risk of hospitalization during infection with SARS-CoV-2. A large number of high quality studies are needed to confirm these results.


Asunto(s)
COVID-19 , Neoplasias de la Próstata , Masculino , Humanos , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/inducido químicamente , Antagonistas de Andrógenos/efectos adversos , Andrógenos/uso terapéutico , SARS-CoV-2
3.
Mol Oncol ; 16(20): 3666-3688, 2022 10.
Artículo en Inglés | MEDLINE | ID: mdl-36052737

RESUMEN

The genetic features of primary lymphoepithelioma-like carcinoma (LELC) of the upper urinary tract have not been systematically explored. In this study, tumor mutation profiling was performed using whole-genome sequencing in two patients with LELC of the renal pelvis. Novel candidate variants relevant to known disease genes were selected using rare-variant burden analysis. Subsequently, a population-based study was performed using the Surveillance, Epidemiology, and End Results (SEER), PubMed, MEDLINE, Embase, and Scopus databases to explore clinical features and prognostic risk factors. Immunohistochemical analysis revealed seven positive cytokeratin-associated markers in tumor cells and five positive lymphocyte-associated markers in and around the tumor area. Sub-sequently, we identified KDM6A as the susceptibility gene and LEPR as the driver gene by Sanger sequencing in case 2 of LELC of the renal pelvis. Three mutation sites of the existing targeted drugs were screened: CA9, a therapeutic target for zonisamide; ARVCF, a therapeutic target for bupropion; and PLOD3, a therapeutic target for vitamin C. In a population-based study, patients with primary LELC of the upper urinary tract had clinical outcomes similar to those of patients with primary upper urinary tract urothelial carcinoma (UUT-UC) before and after propensity score matching at 1 : 5. Focal subtype was an independent prognostic factor for the overall survival of patients with LELC of the upper urinary tract. The carcinogenesis of primary LELC may be due to different genetic variations, including single-nucleotide variants, insertion and deletions, structural variations, and repeat regions, which may provide the basis for clinical diagnosis and treatment. The prognosis of LELC in the upper urinary tract is similar to that of UUT-UC. We suggest that the focal subtype can serve as a prognostic factor for LELC of the upper urinary tract; however, further studies are required to confirm this.


Asunto(s)
Carcinoma de Células Escamosas , Carcinoma de Células Transicionales , Neoplasias de la Vejiga Urinaria , Humanos , Pronóstico , Zonisamida , Bupropión , Pelvis Renal , Histona Demetilasas , Queratinas , Ácido Ascórbico , Nucleótidos
4.
Int J Food Microbiol ; 378: 109805, 2022 Oct 02.
Artículo en Inglés | MEDLINE | ID: mdl-35777301

RESUMEN

Escherichia coli O157 belongs to a diverse serogroup including different H serotypes. E. coli O157: H7 is the most common serotype that can cause acute gastroenteritis, hemorrhagic colitis (HC), and hemolytic-uremic syndrome (HUS) in humans. In recent years, some E. coli O157:non-H7 strains have been reported to cause sporadic cases and outbreaks of diarrheal diseases. However, the phenotypic and genotypic characteristics of E. coli O157:non-H7 are poorly understood. In this study, E. coli O157:non-H7 strains were isolated from retail food sold on markets in 13 cities in China during 2012-2016 and characterized systematically in terms of their H serotypes, virulence genes, antibiotic resistance, and genotypes. Six H serotypes (H26, H42, H11, H38, H4, and H5) were identified, of which, O157:H42 (31.4 %) and O157:H26 (28.6 %) were the most prevalent. Most of the isolates (82.9 %) carried virulence genes. Ten isolates (28.6 %) carried the eae gene and were identified as atypical enteropathogenic E. coli. Multilocus sequence typing showed that the E. coli O157:non-H7 strains demonstrated diverse sequence types with different evolutionary trends than E. coli O157:H7. All the isolates exhibited multidrug resistance. The isolates showed a high prevalence of resistance to AMC, AMP, CTX, CIP, T/S, TE, and FFC. The predominant antibiotic-resistance genes were TEM-1 (40.0 %), CTX-M-55 (34.3 %), aadA (74.3 %), sul2 (62.9 %), floR (91.4 %), tetA (85.7 %), qnrS (37.1 %), oqxA (62.9 %), and oqxB (62.9 %). For the first time, we identified IncI2 plasmid-mediated colistin-resistant strains (six O157:H26 and one O157:H4). These strains co-harbored plasmid-mediated mcr-1 gene, CTX-M-55, OXA-4, PMQR, and other resistant genes, which is of great concern. Colistin and cefotaxime are generally used as the last defense to treat complicated infections. The emergence of virulent and multidrug resistant strains in food poses a serious threat to human health. The strict monitoring and surveillance of multiple-drug resistant strains in food are needed to prevent their dissemination to humans.


Asunto(s)
Escherichia coli Enteropatógena , Infecciones por Escherichia coli , Escherichia coli O157 , Proteínas de Escherichia coli , Colistina/farmacología , Infecciones por Escherichia coli/epidemiología , Proteínas de Escherichia coli/genética , Humanos
5.
Jpn J Clin Oncol ; 52(8): 930-943, 2022 08 05.
Artículo en Inglés | MEDLINE | ID: mdl-35578896

RESUMEN

BACKGROUND: Primary malignant melanoma of the ureter is extremely rare. Genetic variants to the increased risk of developing the disease have not yet been investigated. METHODS: Tumour mutation profiling for primary malignant melanoma of the ureter was performed by whole-exome sequencing. Immunohistochemistry was performed to verify histopathological features and the variants of predisposing genes and driver mutation genes. Furthermore, we conducted a literature review and Surveillance, Epidemiology and End Result-based study by searching public databases. RESULTS: We identified 38 somatic single nucleotide variants and 9 somatic insertions and deletions (INDELs) in tumour specimens. After filtering with the Cancer Gene Census database, seven predisposing genes and two driver mutation genes were identified. Moreover, the immunohistochemical profile showed that tumour cells were positive for Melan-A, melanoma gp100 human melanoma black 45 (HMB45), S100 beta and P53. The expression levels of two driver mutation genes (phosphatase and tensin homolog (PTEN) and desmoyokin (AHNAK) and five predisposing genes (AT-rich interaction domain 1B (ARID1B), catalase, eukaryotic translation initiation factor 4 gamma 3 (EIF4G3), ANK3 and collagen type I) were significantly downregulated in tumour tissues compared to paracancerous tissues. In the literature review and Surveillance, Epidemiology and End Results-based study, patients with primary malignant melanoma of the urinary tract had worse clinical outcomes than patients with primary urothelial carcinoma after 1:2 propensity score matching (P = 0.010). Additionally, Cox multivariate analysis for patients with primary malignant melanoma of the urinary tract indicated that distant metastasis (hazard ratio = 1.185; P = 0.044) was an independent predictor for overall survival, and tumour focality (hazard ratio = 0.602; P = 0.017) and non-surgery (hazard ratio = 0.434; P = 0.003) were independent factors for tumour progression. CONCLUSIONS: Our study is the first to provide evidence that the distinct phenotypes of primary malignant melanoma of the ureter may be due to different genetic variations. The prognosis of primary malignant melanoma of the urinary tract was poorer than that of primary urothelial carcinoma of the urinary tract.


Asunto(s)
Carcinoma de Células Transicionales , Melanoma , Proteínas de la Membrana , Proteínas de Neoplasias , Fosfohidrolasa PTEN , Neoplasias de la Vejiga Urinaria , Carcinoma de Células Transicionales/genética , Carcinoma de Células Transicionales/patología , Genómica , Humanos , Melanoma/genética , Melanoma/patología , Proteínas de la Membrana/genética , Mutación , Proteínas de Neoplasias/genética , Fosfohidrolasa PTEN/genética , Uréter/cirugía , Neoplasias de la Vejiga Urinaria/genética , Neoplasias de la Vejiga Urinaria/patología
6.
Front Oncol ; 11: 731318, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34660295

RESUMEN

BACKGROUND: Upper urinary tract urothelial carcinoma (UUT-UC) is a rare and severe urinary malignancy. Several studies have explored the relationship between preoperative urine cytology and intravesical recurrence (IVR) in patients with UUT-UC. However, the results of these studies are controversial or even contradictory, and investigations with UUT-UC patients in northeast China are rare. METHODS: We first estimated the prognostic significance of preoperative urine cytology in the outcomes of intravesical recurrence in 231 UUT-UC patients (training cohort = 142, validation cohort = 89) after radical nephroureterectomy (RNU) by the nomogram model. Subsequently, we quantitatively combined our results with the published data after searching several databases to assess whether preoperative positive urine cytology was associated with poor intravesical recurrence-free survival and a high risk of tumor malignant biological behavior. RESULTS: Firstly, the multicenter retrospective cohort study demonstrated that preoperative positive urine cytology correlated with poor intravesical recurrence-free survival and can serve as significant independent predictors of IVR by Kaplan-Meier curves and Cox regression analysis. The construction of the nomogram demonstrated that predictive efficacy and accuracy were significantly improved when preoperative urine cytology was combined. Meanwhile, meta-analysis showed that preoperative positive urine cytology was associated with a 49% increased risk of IVR. In the subgroup analysis by region, study type, and sample size, the pooled hazard ratios (HRs) were statistically significant for the Japan subgroup (HR 1.32), China subgroup (HR 1.88), cohort study subgroup (HR 1.45), and the single-arm study subgroup (HR 1.63). CONCLUSIONS: Preoperative urine cytology was validated as a potential predictor of intravesical recurrence in patients with UUT-UC after RNU, although these results need to be generalized with caution. Large, prospective trials are required to further confirm its significance in prognosis and tumor malignant biological behavior.

7.
Front Oncol ; 11: 633462, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34350107

RESUMEN

Aspirin, widely used to prevent cardiovascular disease, had been linked to the incidence of bladder cancer (BCa). Existing studies focusing on Chinese populations are relatively rare, especially for Northeast China. Meanwhile, relevant studies on the effects of aspirin on the occurrence or prognosis of BCa are inconsistent or even controversial. First, in the case control study, logistic regression analysis was used to investigate the association between aspirin intake and risk of BCa including 1121 patients with BCa and the 2242 controls. Subsequently, Kaplan-Meier curve and Cox regression analyses were applied to explore the association between aspirin intake and clinicopathological factors which may predict overall survival (OS) and recurrence-free survival (RFS) of BCa patients. Finally, we quantificationally combined the results with those from the published literature evaluating aspirin intake and its effects on the occurrence, outcome of surgery and prognosis of BCa by meta-analysis up to May 1, 2021.Our case-control study demonstrated that the regular use of aspirin was not associated with a reduced incidence of BCa (P=0.175). Stratified analyses of sex showed that aspirin intake did not lead to a lower risk of BCa in female patients (P=0.063). However, the male population who regularly took aspirin had a lower incidence of BCa (OR=0.748, 95% CI= 0.584-0.958, P=0.021). Subgroup analyses stratified by smoking found a significant reduction in the risk of BCa in current smokers with aspirin intake (OR=0.522, 95% CI=0.342-0.797, P=0.002). In terms of prognosis of BCa, patients with a history of aspirin intake did not had a markedly longer OS or RFS than those with no history of aspirin intake by Kaplan-Meier curves. Stratified analysis by sex showed no correlation between aspirin intake and the recurrence or survival of BCa for either male or female patients. However, in people younger than 68, aspirin intake seemed to have prolonged effects for overall survival (HR=3.876; 95% CI=1.326-11.325, P=0.019). Then, we performed a meta-analysis and the combined results from 19 articles and our study involving more than 39524 BCa cases indicated that aspirin intake was not associated with the occurrence of BCa (P=0.671). Subgroup analysis by whether regular use of aspirin, by the mean duration of use of aspirin, by sex, by smoking exposure, by research region and by study type also supported the above results. In terms of the impact of aspirin intake on the prognosis of patients with BCa, 11 articles and our study involving 8825 BCa cases were eligible. The combined results showed that patients with aspirin intake did not have significantly influence on survival, recurrence, progression and metastasis than those without aspirin intake. On the whole, both our retrospective study and literature meta-analysis suggested a lack of a strong relevant association between the use of aspirin and the incidence or prognosis of BCa. Thus, additional long-term follow-up prospective research is warranted to clarify the association of aspirin with BCa incidence and prognosis.

8.
Int J Food Microbiol ; 355: 109349, 2021 Oct 02.
Artículo en Inglés | MEDLINE | ID: mdl-34371389

RESUMEN

Antibiotic-resistant bacteria in food pose an important threat to public health. Multidrug-resistant strains in ready-to-eat (RTE) foods can be transferred to humans through diet, which increases their health risk. This study systematically investigated antibiotic resistance and antibiotic resistance genes in E. coli isolated from retail RTE foods and characterized plasmid-mediated colistin-resistant E. coli strains. A total of 1118 RTE food samples were collected from markets in 39 cities in China, and 126 E. coli strains, >95% of which were multidrug-resistant, were isolated. The isolates showed a high prevalence of resistance to tetracycline (95.24%), ampicillin (82.54%), trimethoprim-sulfamethoxazole (77.78%), nalidixic acid (74.60%), cephalothin (72.22%), chloramphenicol (66.67%), and streptomycin (53.97%). Twenty-two extended-spectrum ß-lactamase (ESBL)-producing E. coli and four colistin-resistant E. coli were identified. The resistance genes TEM, CTX-M, tetA, sul2, strA/strB, aadA, and qnrS were the most frequently detected. CTX-M-55 and CTX-M-14 were the predominant CTX-M types. All the four colistin-resistant E. coli isolates were positive for mcr-1. The mcr-1 gene can be transferred to E. coli C600 through conjugation and transformation. Whole-genome sequencing revealed that the mcr-1 genes were found in IncX4 and IncHI2 plasmids. To the best of our knowledge, this is the first report of IncHI2/IncX4 plasmid-bearing mcr-1-positive E. coli strains in RTE foods sold in markets, and the first report of the isolation of the international epidemic E. coli clone ST101 and mcr-1-carrying ESBL-producing E. coli from RTE foods. These results provide valuable information for assessing antibiotic-resistant E. coli infections and controlling antibiotic-resistant E. coli.


Asunto(s)
Farmacorresistencia Bacteriana Múltiple , Infecciones por Escherichia coli , Proteínas de Escherichia coli , Escherichia coli , Comida Rápida , Microbiología de Alimentos , Antibacterianos/farmacología , Farmacorresistencia Bacteriana Múltiple/genética , Escherichia coli/genética , Escherichia coli/aislamiento & purificación , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/microbiología , Proteínas de Escherichia coli/genética , Comida Rápida/microbiología , Humanos , Pruebas de Sensibilidad Microbiana , Plásmidos/genética , Prevalencia , beta-Lactamasas/genética
9.
J Agric Food Chem ; 69(28): 7938-7947, 2021 Jul 21.
Artículo en Inglés | MEDLINE | ID: mdl-34237214

RESUMEN

A novel cell-free biosynthesis system based on a mixture of chassis cell extracts and purified Spy-cyclized enzymes (CFBS-mixture) was developed. As a demonstration, the CFBS-mixture was applied to chlorogenic acid (CGA) biosynthesis. The mix-and-match and Plackett-Burman experiments demonstrated that Lonicera japonica hydroxycinnamate-CoA quinate transferase and p-hydroxyphenylacetate 3-hydroxylase were the key enzymes for the production of CGA. After optimization of the concentrations of the biosynthetic enzymes in the CFBS-mixture reaction using the Plackett-Burman experimental design and the path of the steepest ascent, 711.26 ± 15.63 mg/L CGA was produced after 16 h, which is 71.1-fold the yield obtained using the conventional crude extract-based CFBS and 9.1-fold the reported yield obtained using the living cells. Based on the CFBS-mixture results, the production of CGA was further enhanced in engineered Escherichia coli. The CFBS-mixture strategy is highly effective and will be useful for high-level CFBS of natural products.


Asunto(s)
Ácido Clorogénico , Lonicera , Extractos Celulares , Ácido Quínico
10.
Virus Res ; 293: 198274, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33359502

RESUMEN

Phage therapy has been revitalized since antibiotic resistance in bacteria is increasing. Compared with antibiotics, phages can target specific bacteria precisely, which requires more understanding of phage-host interactions by investigating different phages. Escherichia coli is a common pathogen with very high diversity. Based on the O antigens, E. coli can be classified into at least 183 serotypes and existing phages are far from being able to lyse all E. coli. Therefore, a novel phage specific to E. coli, named DY1, was identified and characterized to enhance our understanding of the phages of E. coli and expand the phage library. Phage DY1 belongs to the family Autographiviridae which is derived from Podoviridae. The genome of DY1 was determined to be 39,817 bp and comprises 54 putative open reading frames. Comparative genome and phylogenetic analysis demonstrated that DY1 was highly similar to phages belonging to the genus Kayfunavirus; however, the highest average nucleotide identity (ANI) values of DY1 with known phages was 0.82 suggesting that DY1 was a novel phage. Through stability tests, DY1 was very stable at temperatures ranging from 20 to 50 °C and pH levels from 5 to 11. Taken together, we report that phage DY1 is a novel Kayfunavirus phage with the potential for phage therapy.


Asunto(s)
Bacteriófagos , Escherichia coli , Escherichia coli/genética , Genómica , Filogenia
11.
J Agric Food Chem ; 68(42): 11765-11773, 2020 Oct 21.
Artículo en Inglés | MEDLINE | ID: mdl-33030899

RESUMEN

Here, we first developed a combined strain improvement strategy of biosensor-guided atmospheric and room-temperature plasma mutagenesis and genome shuffling. Application of this strategy resulted in a 2.7-fold increase in the production of shikimic acid (SA) and a 2.0-fold increase in growth relative to those of the starting strain. Whole-cell resequencing of the shuffled strain and confirmation using CRISPRa/CRISPRi revealed that some membrane protein-related mutant genes are identified as being closely related to the higher SA titer. The engineered shuffling strain produced 18.58 ± 0.56 g/L SA from glucose with a yield of 68% (mol/mol) by fed-batch whole-cell biocatalysis, achieving 79% of the theoretical maximum. Sucrose-utilizing Escherichia coli was engineered for SA production by introducing Mannheimia succiniciproducens ß-fructofuranosidase gene. The resulting sucrose-utilizing E. coli strain produced 24.64 ± 0.32 g/L SA from sucrose with a yield of 1.42 mol/mol by fed-batch whole-cell biocatalysis, achieving 83% of the theoretical maximum.


Asunto(s)
Escherichia coli/genética , Escherichia coli/metabolismo , Ácido Shikímico/metabolismo , Sacarosa/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Barajamiento de ADN , Ingeniería Metabólica , Mutagénesis , Pasteurellaceae/enzimología , Pasteurellaceae/genética , beta-Fructofuranosidasa/genética , beta-Fructofuranosidasa/metabolismo
12.
Artículo en Inglés | MEDLINE | ID: mdl-32432104

RESUMEN

Aromatic compounds derived from aromatic amino acids are an important class of diverse chemicals with a wide range of industrial and commercial applications. They are currently produced via petrochemical processes, which are not sustainable and eco-friendly. In the past decades, significant progress has been made in the construction of microbial cell factories capable of effectively converting renewable carbon sources into value-added aromatics. Here, we systematically and comprehensively review the recent advancements in metabolic engineering and synthetic biology in the microbial production of aromatic amino acid derivatives, stilbenes, and benzylisoquinoline alkaloids. The future outlook concerning the engineering of microbial cell factories for the production of aromatic compounds is also discussed.

13.
J Food Prot ; 83(3): 460-466, 2020 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-32065650

RESUMEN

ABSTRACT: Non-O157 Shiga toxin-producing Escherichia coli (STEC) strains are significant foodborne pathogens that can cause acute diarrhea in humans. This study was conducted to investigate the contamination by non-O157 STEC in different types of food sold at retail markets in the People's Republic of China and to characterize non-O157 STEC strains. From May 2012 to April 2014, 1,200 retail food samples were collected from markets in 24 cities in China. Forty-four non-O157 isolates were recovered from 43 STEC-positive samples. Of the isolates, 22 and 19 carried the stx1 and stx2 genes, respectively, and 3 harbored both stx1 and stx2. stx1a and stx2a were the most prevalent stx subtypes. Other virulence genes, ent, hlyA, astA, eae, espB, iha, subAB, and tia, were commonly detected. Diverse O serogroups were identified among these isolates. Multilocus sequence typing indicated the high genetic diversity of the isolates. Thirty-two sequence types (STs) were identified among the 44 isolates, with ST383 (9.09%), ST134 (6.82%), and ST91 (6.82%) the most prevalent. Nine new STs were found. The isolates had a high prevalence of resistance to cephalothin, ampicillin, tetracycline, trimethoprim-sulfamethoxazole, nalidixic acid, streptomycin, and chloramphenicol. Twenty isolates (45.45%) were resistant to at least three antibiotics. This study provides updated surveillance data for non-O157 STEC isolates from foods sold at retail markets. Virulent and multidrug-resistant non-O57 STEC strains were isolated from all types of food. Our findings highlight the need for increased monitoring of non-O157 STEC in retail foods.


Asunto(s)
Proteínas de Escherichia coli , Contaminación de Alimentos/análisis , Escherichia coli Shiga-Toxigénica , China , Proteínas de Escherichia coli/genética , Microbiología de Alimentos , Humanos , Serogrupo , Escherichia coli Shiga-Toxigénica/clasificación , Escherichia coli Shiga-Toxigénica/aislamiento & purificación
14.
J Agric Food Chem ; 68(7): 2139-2145, 2020 Feb 19.
Artículo en Inglés | MEDLINE | ID: mdl-31973519

RESUMEN

α-Pinene is an important monoterpene that is widely used as a pharmaceutical product, biofuel, and so forth. We first established a cell-free system with modular cocatalysis for the production of pinene from glucose. After optimization of the compositions of the cell-free reaction mixture using the Plackett-Burman experimental design and the path of steepest ascent, the production of pinene increased by 57%. It was found that ammonium acetate, NAD+, and NADPH are the three most important parameters for the production of pinene. Mix-and-match experiments showed that the simultaneous addition of the lysate of Escherichia coli overexpressing native 4-hydroxy-3-methylbut-2-enyl diphosphate reductase, SufBCD Fe-S cluster assembly protein, isopentenyl-diphosphate isomerase, and Pinus taeda pinene synthase improved the production of pinene. Increasing the enzyme concentration of the extract further enhanced the production of pinene to 1256.31 ± 46.12 mg/L with a productivity of 104.7 mg/L h, almost 1.2-fold faster than any system reported thus far. This study demonstrates that a cell-free system is a powerful and robust platform for biomanufacture.


Asunto(s)
Monoterpenos Bicíclicos/química , Escherichia coli/química , Monoterpenos Bicíclicos/metabolismo , Catálisis , Escherichia coli/genética , Escherichia coli/metabolismo , Cinética , NAD/química , NAD/metabolismo , NADP/química , NADP/metabolismo
15.
Synth Syst Biotechnol ; 4(3): 113-119, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31198860

RESUMEN

α-Pinene is an important monoterpene, which is widely used as a flavoring agent and in fragrances, pharmaceuticals and biofuels. Although an evolved strain Escherichia coli YZFP, which had higher tolerance to pinene and titer, has been successfully used to produce high levels of pinene, the pinene titer is much lower than that of hemiterpene (isoprene) and sesquiterpenes (farnesene) to date. Moreover, the overall cellular physiological and metabolic changes caused by higher tolerance to pinene and overproduction of pinene remains unclear. To reveal the mechanism of Escherichia coli YZFP with the higher tolerance to pinene and titer, a comparative genomics and transcriptional level analyses combining with CRISPR activation (CRISPRa) and interference (CRISPRi) were carried out. The results show that the tolerance to pinene and the overproduction of pinene in E. coli may be associated with: 1) the mutations of the DXP pathway genes, the rpoA and some membrane protein genes, and their upregulations of transcription levels; and 2) the mutations of some genes and their downregulation of transcriptional levels. These comparative omics analyses provided some genetic modification strategies to further improve pinene production. Overexpression of the mutated cbpA, tabA, pitA, rpoA, sufBCDS, mutS, ispH, oppF, dusB, dnaK, dxs, dxr and flgFGH genes further improved pinene production. This study also demonstrated that combining comparative omics analysis with CRISPRa and CRISPRi is an efficient technology to quickly find a new metabolic engineering strategy.

16.
J Food Prot ; 81(11): 1761-1767, 2018 11.
Artículo en Inglés | MEDLINE | ID: mdl-30277802

RESUMEN

Atypical enteropathogenic Escherichia coli (aEPEC) is an emerging pathogen that has been implicated in outbreaks of diarrhea worldwide. The objective of this study was to investigate the occurrence of aEPEC in retail foods at markets in the People's Republic of China and to characterize the isolates for virulence genes, intimin gene ( eae) subtypes, multilocus sequence types (STs), and antimicrobial susceptibility. From May 2014 to April 2015, 1,200 food samples were collected from retail markets in China, and 41 aEPEC isolates were detected in 2.75% (33 of 1,200) of the food samples. The virulence genes tir, katP, etpD, efa/lifA, ent, nleB, and nleE were commonly detected in these isolates. Nine eae subtypes were detected in the isolates, among which θ (23 isolates) and ß1 (6 isolates) were the most prevalent. The 41 isolates were divided into 27 STs by multilocus sequence typing. ST752 and ST10 were the most prevalent. Antibiotic susceptibility testing revealed high resistance among isolates to streptomycin (87.80%), cephalothin (73.16%), ampicillin (51.22%), tetracycline (63.42%), trimethoprim-sulfamethoxazole (43.90%), and kanamycin (43.90%). Thirty isolates (73.17%) were resistant to at least three antibiotics, and 20 (53.66 %) were resistant to five or more antibiotics. Our results suggest that retail foods in markets are important sources of aEPEC. The presence of virulent and multidrug-resistant aEPEC in retail foods poses a potential threat to consumers. Surveillance of aEPEC contamination and prudent use of antibiotics is strongly recommended in China.


Asunto(s)
Escherichia coli Enteropatógena , Comida Rápida/microbiología , Contaminación de Alimentos/análisis , Microbiología de Alimentos , Antibacterianos/farmacología , China , Diarrea/epidemiología , Diarrea/microbiología , Escherichia coli Enteropatógena/efectos de los fármacos , Escherichia coli Enteropatógena/genética , Escherichia coli Enteropatógena/aislamiento & purificación , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/microbiología , Humanos , Prevalencia , Virulencia
17.
Front Microbiol ; 9: 289, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29545778

RESUMEN

Klebsiella pneumoniae is not only a major hospital-acquired pathogen but also an important food-borne pathogen that can cause septicaemia, liver abscesses, and diarrhea in humans. The phenotypic and genotypic characteristics of K. pneumoniae in retail foods have not been thoroughly investigated in China. The objective of this study was to characterize K. pneumoniae isolates through biotyping, serotyping, determination of virulence factors, antibiotic resistance testing, enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR), and (GTG)5-PCR molecular typing. From May 2013 to April 2014, a total of 61 K. pneumoniae isolates were collected from retail foods in China. Using API 20E test strips, five different biotype profiles were identified among these isolates. The majority of isolates belonged to biochemical profile "5215773" (50 isolates, 80.6%). The capsular serotypes of the 61 K. pneumoniae isolates and one reference strain were determined by PCR. Of the seven capsular serotypes tested, four different capsular serotypes were identified. Serotypes K1, K20, K57, and K2 were detected in two, three, two, and one isolates, respectively. Serotypes K3, K5, and K54 were not detected. The presence of 11 virulence genes was assessed by PCR. The most common virulence genes were fimH (85.5%), ureA (79.0%), wabG (77.4%), uge (56.5%), and kfuBC (29.0%). ERIC-PCR and (GTG)5-PCR molecular typing indicated high genetic diversity among K. pneumoniae isolates. We identified 60 different ERIC patterns and 56 distinct (GTG)5 patterns. Genotypic results indicated that isolates carrying similar virulence factors were generally genetically related. Some isolates from the same geographic area have a closer relationship. The isolates showed high levels of resistance to ampicillin (51/62, 82.2%). Resistance to streptomycin (11/62, 17.7%) and piperacillin (10/62, 16.1%) was also common. The presence of virulent and antibiotic-resistant K. pneumoniae in foods poses a potential health hazard for consumers. Our findings highlight the importance of surveillance of K. pneumoniae in foods.

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