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1.
Crit Rev Food Sci Nutr ; : 1-20, 2024 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-38881345

RESUMO

Inflammation is a complex process that usually refers to the general response of the body to the harmful stimuli of various pathogens, tissue damage, or exogenous pollutants. Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor that regulates cellular defense against oxidative damage and toxicity by expressing genes related to oxidative stress response and drug detoxification. In addition to its antioxidant properties, Nrf2 is involved in many other important physiological processes, including inflammation and metabolism. Nrf2 can bind the promoters of antioxidant genes and upregulates their expressions, which alleviate oxidation-induced inflammation. Nrf2 has been shown to upregulate heme oxygenase-1 expression, which promotes NF-κB activation and is closely related with inflammation. Nrf2, as a key factor in antioxidant response, is closely related to the expressions of pro-inflammatory factors, NF-κB pathway and cell metabolism. Bioactive peptides come from a wide range of sources and have many biological functions. Increasing evidence indicates that bioactive peptides have potential anti-inflammatory activities. This article summarized the sources, absorption and utilization of bioactive peptides and their role in alleviating inflammation via Nrf2 pathway. Bioactive peptides can also regulate gut microbiota and alter metabolites, which regulates the Nrf2 pathway through novel pathway and supplement the anti-inflammatory mechanisms of bioactive peptides. This review provides a reference for further study on the anti-inflammatory effect of bioactive peptides and the development and utilization of functional foods.

2.
Artigo em Inglês | MEDLINE | ID: mdl-39073850

RESUMO

A Gram-stain-positive, rod-shaped, non-spore-forming and non-motile bacterium, designated strain WY-16T. Growth was observed at 20-42 °C (optimum, 30 °C), pH 6-9 (optimum, pH 7) and salinity of 0-3 % (w/v; optimum, 1 %). Phylogenetic analysis based on genome sequences indicated that WY-16T was affiliated to the family Microbacteriaceae and most closely related to Salinibacterium xinjiangense and Salinibacterium amurskyense. The average nucleotide identity values between strain WY-16T and S. xinjiangense and S. amurskyense were 74.7 and 72.5 %, respectively. The digital DNA-DNA hybridization values between strain WY-16T and S. xinjiangense and S. amurskyense were 19.6 and 18.6 %, respectively. The predominant fatty acids were anteiso-C15 : 0, iso-C16 : 0 and iso-C16 : 0 10-methyl. The major menaquinones were MK-12, MK-13, MK-14 and MK-15. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, one unidentified glycolipid and one unidentified phospholipid. The cell-wall peptidoglycan contained 2,4-diaminobutyric acid as the diamino acid and ribose, rhamnose, glucose and galactose were the major cell-wall sugars. Based on phenotypic, genotypic and phylogenetic evidence, strain WY-16T represents a novel species in the genus Salinibacterium, for which the name Salinibacterium soli sp. nov. is proposed. The type strain is WY-16T (=GDMCC 1.4011T=JCM 36421T).


Assuntos
Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano , Ácidos Graxos , Lagos , Hibridização de Ácido Nucleico , Fosfolipídeos , Filogenia , RNA Ribossômico 16S , Análise de Sequência de DNA , Microbiologia do Solo , Vitamina K 2 , Ácidos Graxos/química , Ácidos Graxos/análise , RNA Ribossômico 16S/genética , Vitamina K 2/análogos & derivados , Vitamina K 2/análise , DNA Bacteriano/genética , Fosfolipídeos/química , Fosfolipídeos/análise , Lagos/microbiologia , Peptidoglicano , China
3.
Environ Sci Technol ; 2024 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-39096310

RESUMO

Polyhalogenated dibenzo-p-dioxins/dibenzofurans (PXDD/Fs) are commonly released into the environment as byproducts of combustion processes, accompanied by flue gases. Chlorinated (Cl) and brominated (Br) precursors play crucial roles in forming PXDD/Fs. However, the specific contributions of Cl-precursors and Br-precursors to PXDD/Fs formation have not been fully elucidated. Herein, we demonstrate that the formation of Br-precursors can increase the fraction of polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs) congeners substituted at specific positions, such as 1,2,3,4,6,7,8-HpCDD, OCDD, 2,3,4,7,8-PeCDF, and 2,3,4,6,7,8-HxCDF. This is attributed to the electrophilic chlorination reaction of the Br-precursors, which includes the Br-to-Cl transformation pathway, following the principle of regioselectivity. The observed formation of polybrominated/chlorinated dibenzo-p-dioxins/benzofurans (PBCDD/Fs) from 1,2-dibromobenzene (1,2-DiBBz) as a Br precursor provides direct evidence supporting the proposed Br-to-Cl transformation. Quantum chemical calculations are employed to discuss the principle of regioselectivity in the Br-to-Cl transformation, clarifying the priority of the position for electrophilic chlorination. Additionally, the concentration of PCDD/Fs formed from 1,2-DiBBz is 1.6 µg/kg, comparable to that of polybrominated dibenzo-p-dioxins/dibenzofurans (PBDD/Fs) (2.4 µg/kg), highlighting the potential of brominated organic pollutants as precursors for PCDD/Fs formation. This study provides three potential pathways for PCDD/Fs formation from Br-precursors, establishing a theoretical foundation for elucidating the formation mechanism of PXDD/Fs in the coexistence of Cl and Br.

4.
Clin Invest Med ; 47(3): 7-17, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39325577

RESUMO

PURPOSE: This study aimed to establish a CT imaging grading system and explore its value in evaluating upper urinary tract calculi associated with kidney infections. METHODS: CT images of 126 patients with kidney infections caused by upper urinary tract calculi were retrospectively analyzed. The CT grading system was developed based on CT images. CT images were classified into 4 grades. General information, symptoms, and clinical findings of patients in different CT grades were analyzed. With the occurrence of systemic inflammatory response syndrome (SIRS) as the endpoint, univariate and multivariate analysis was conducted to analyze the risk factors of SIRS. RESULTS: Patients with fever or diabetes had higher CT grades, and the following examination data revealed significant differences across the various CT grades (P < 0.05): the white blood cell count, urine leucocytes count, CT1, CT2, maximum body temperature, duration of disease, the proportion of blood neutrophils, the size of stones, and levels of the C-reactive protein and procalcitonin. Only CT grading was statistically significant after multivariate analysis. According to the values of the partial regression coefficient (B), the higher the CT grade, the greater the risk of SIRS. The risk of SIRS was 4.472 times higher with each increment of the CT grade. CONCLUSIONS: The CT grade is directly associated with clinical symptoms and the risk of SIRS.


Assuntos
Tomografia Computadorizada por Raios X , Infecções Urinárias , Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Adulto , Infecções Urinárias/diagnóstico por imagem , Idoso , Estudos Retrospectivos , Síndrome de Resposta Inflamatória Sistêmica/diagnóstico por imagem , Cálculos Urinários/diagnóstico por imagem , Cálculos Urinários/complicações
5.
BMC Nurs ; 23(1): 256, 2024 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-38649865

RESUMO

BACKGROUND: Abusive supervision by the nurse manager significantly influences nurses' withholding voice about patient safety. The role of impression management motivation and speak up-related climate is crucial in understanding their connection. This study aimed to explore the relationship between abusive supervision, impression management motivation, speak up-related climate, and withholding voice about patient safety. METHODS: This cross-sectional study employed a convenience sampling method to recruit 419 clinical nurses from Taizhou Hospital, Zhejiang Province, China, between 1 November 2022 and 31 January 2023. The study adhered to the STROBE checklist. Abusive supervision and impression management motivation were assessed using the Chinese versions of the Abusive Supervision Scale and the Impression Management Motivation Scale, respectively. Withholding voice about patient safety and speak up-related climate were identified using the Chinese version of the Speaking Up about Patient Safety Questionnaire. RESULTS: Nurse leaders' abusive supervision (ß=0.40, p<0.01) and nurses' impression management motivation (ß=0.10, p<0.01) significantly and positively influenced nurses' withholding voice about patient safety. We introduced impression management motivation as a mediating variable, and the effect of abusive supervision on nurses' withholding voice decreased (ß from 0.40 to 0.38, p< 0.01). Nurses' speak up-related climate played a moderating role between abusive supervision and impression management motivation (ß= 0.24, p<0.05). CONCLUSIONS: Abusive supervision by nursing leaders can result in nurses withholding voice about patient safety out of self-protective impression management motives. This phenomenon inhibits nurses' subjective initiative and undermines their proactive involvement in improving patient safety, and hinders the cultivation of a culture encouraging full participation in patient safety, which should warrant significant attention.

6.
Minim Invasive Ther Allied Technol ; 33(1): 29-34, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37971312

RESUMO

OBJECTIVE: To describe our technique of transvesical laparoscopic simple prostatectomy (LSP) plus complete urethral reconstruction(CUR). MATERIAL AND METHODS: From May 2019 to May 2021, 28 BPH patients with prostate volumes > 80 ml and the requirement to preserve the ejaculatory function (EF) received LSP plus CUR. Baseline demographics, pathology data, perioperative and postoperative complications, and functional outcomes were assessed. Data were analyzed with the Wilcoxon test. RESULTS: The median prostate volume was 106 ml. All patients successfully underwent LSP with no intraoperative complications or conversions to open surgery. The median operative time was 146 min. A total of five Clavien-Dindo Grade1-2 postoperative complications were noted, including infection, prolonged urine leakage and cardiac arrhythmia. No patient reported postoperative urgent or stress urinary incontinence. Functional outcomes at one-year follow-up demonstrated significant improvement from baseline with median IPSS and Qmax (p both < 0.001). Compared with baseline, no significant difference was observed in IIEF and MSHQ-EjD-SF at 6 and 12 months postoperatively. CONCLUSIONS: Our data support transperitoneal-transvesical LSP plus CUR as a safe and effective surgical technique for treating BPH with large prostate adenoma, regardless of the volume of the median lobe, especially for patients requiring to preserve antegrade ejaculation.


Assuntos
Laparoscopia , Hiperplasia Prostática , Masculino , Humanos , Hiperplasia Prostática/cirurgia , Resultado do Tratamento , Prostatectomia/métodos , Complicações Pós-Operatórias/epidemiologia
7.
Minim Invasive Ther Allied Technol ; 33(1): 51-57, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38147882

RESUMO

OBJECTIVES: To compare the effect and safety of retroperitoneal laparoscopic pyelolithotomy (RLP) and percutaneous nephrolithotomy (PCNL) for large pelvis calculi with chronic kidney disease (CKD). MATERIAL AND METHODS: Between June 2017 and July 2021, 62 patients with CKD and large renal pelvis calculi (>4 cm2) were treated with RLP. Another 62 patients receiving PCNL served as controls. The perioperative parameters were compared. All patients were followed up for at least 6 months with the stone-free rate and the recovery of renal function evaluated. RESULTS: Significantly longer operation time (101.47 ± 9.25 vs 62.55 ± 7.54 min), less drop in hemoglobin level (0.90 ± 0.38 vs 2.13 ± 0.80 g/dl), staged operations (0% vs 12.9%), postoperative fever (3.23% vs 16.13%) and delayed bowel movement (3.23% vs 14.52), and shorter hospitalization time (3.90 ± 1.66 vs 4.72 ± 1.80 days) were observed in the RLP group (p < 0.05). The stone-free rates were 100% in the RLP group and 88.7% in the PCNL group at the 3-months follow-up (p < 0.05). The serum creatinine level was significantly lower in the RLP group at 24 h (2.81 ± 1.18 vs 3.00 ± 1.15 mg/dl) and 1 week (2.08 ± 1.13 vs 2.34 ± 1.01 mg/dl) postoperatively (p < 0.05). CONCLUSIONS: Although associated with a longer operation time, RLP is a safer and more efficient surgical option for CKD patients with large pelvic stones than PCNL.


Assuntos
Cálculos Renais , Laparoscopia , Nefrolitotomia Percutânea , Nefrostomia Percutânea , Insuficiência Renal Crônica , Humanos , Laparoscopia/efeitos adversos , Nefrostomia Percutânea/efeitos adversos , Resultado do Tratamento , Cálculos Renais/cirurgia , Insuficiência Renal Crônica/etiologia , Insuficiência Renal Crônica/cirurgia , Estudos Retrospectivos
8.
Angew Chem Int Ed Engl ; : e202412735, 2024 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-39205491

RESUMO

The rapid transport kinetics of divalent magnesium ions are crucial for achieving distinguished performance in aqueous magnesium-ion battery-based energy storage capacitors. However, the strong electrostatic interaction between Mg2+ with double charges and the host material significantly restricts Mg2+ diffusivity. In this study, a new composite material, EDA-Mn2O3, with double-energy storage mechanisms comprising an organic phase (ethylenediamine, EDA) and an inorganic phase (manganese sesquioxide) was successfully synthesized via an organic-inorganic coupling strategy. Inorganic-phase Mn2O3 serves as a scaffold structure, enabling the stable and reversible intercalation/deintercalation of magnesium ions. The organic phase EDA adsorbed onto the surface of Mn2O3 as an elastic matrix, works synergistically with Mn2O3, and utilizes bidentate chelating ligands to capture Mg2+. The robust coordination effect of terminal biprotonic amine in EDA enhances the structural diversity and specific capacity characteristics of the composite material, as further corroborated by density functional theory (DFT) calculations, ex-situ XRD, XPS, and Raman spectroscopy. As expected, an aqueous magnesium ion capacitor with EDA-Mn2O3 serving as the cathode can reach 110.17 Wh/kg. This study aimed to explore the practical application value of organic‒inorganic composite electrodes with double-energy storage mechanisms.

9.
Crit Rev Food Sci Nutr ; : 1-22, 2023 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-37300491

RESUMO

Aging-related cognitive impairment, mainly Alzheimer's disease (AD), has been widely studied. However, effective prevention and treatment methods are still lacking. In recent years, researchers have observed beneficial effects of plant-based supplements, such as flavonoids, on cognitive protection. This provides a new clue for the prevention of cognitive dysfunction. Studies have shown that dietary flavonoids have neuroprotective effects, but the mechanism is not clear. In this review, we systematically reviewed the research progress on the effects of dietary flavonoids on gut microbes and their metabolites, and concluded that flavonoids could improve cognitive function through the gut-brain axis. Flavonoids can be absorbed through the intestine, cross the blood-brain barrier, and enter the brain tissue. Flavonoids can inhibit the expression and secretion of inflammatory factors in brain tissue, reduce the damage caused by oxidative stress, clear neural damage proteins and inhibit neuronal apoptosis, thereby ameliorating age-related cognitive disorders. Future work will continue to explore the gut-brain axis and target genes regulated by flavonoids. In addition, clinical research and its mechanisms need to be further explored to provide solutions or advise for patients with cognitive impairment.

10.
J Med Internet Res ; 24(8): e38082, 2022 08 09.
Artigo em Inglês | MEDLINE | ID: mdl-35943767

RESUMO

BACKGROUND: Heart failure (HF) is a common disease and a major public health problem. HF mortality prediction is critical for developing individualized prevention and treatment plans. However, due to their lack of interpretability, most HF mortality prediction models have not yet reached clinical practice. OBJECTIVE: We aimed to develop an interpretable model to predict the mortality risk for patients with HF in intensive care units (ICUs) and used the SHapley Additive exPlanation (SHAP) method to explain the extreme gradient boosting (XGBoost) model and explore prognostic factors for HF. METHODS: In this retrospective cohort study, we achieved model development and performance comparison on the eICU Collaborative Research Database (eICU-CRD). We extracted data during the first 24 hours of each ICU admission, and the data set was randomly divided, with 70% used for model training and 30% used for model validation. The prediction performance of the XGBoost model was compared with three other machine learning models by the area under the curve. We used the SHAP method to explain the XGBoost model. RESULTS: A total of 2798 eligible patients with HF were included in the final cohort for this study. The observed in-hospital mortality of patients with HF was 9.97%. Comparatively, the XGBoost model had the highest predictive performance among four models with an area under the curve (AUC) of 0.824 (95% CI 0.7766-0.8708), whereas support vector machine had the poorest generalization ability (AUC=0.701, 95% CI 0.6433-0.7582). The decision curve showed that the net benefit of the XGBoost model surpassed those of other machine learning models at 10%~28% threshold probabilities. The SHAP method reveals the top 20 predictors of HF according to the importance ranking, and the average of the blood urea nitrogen was recognized as the most important predictor variable. CONCLUSIONS: The interpretable predictive model helps physicians more accurately predict the mortality risk in ICU patients with HF, and therefore, provides better treatment plans and optimal resource allocation for their patients. In addition, the interpretable framework can increase the transparency of the model and facilitate understanding the reliability of the predictive model for the physicians.


Assuntos
Insuficiência Cardíaca , Aprendizado de Máquina , Estudos de Coortes , Insuficiência Cardíaca/terapia , Humanos , Unidades de Terapia Intensiva , Reprodutibilidade dos Testes , Estudos Retrospectivos
11.
Appl Microbiol Biotechnol ; 104(12): 5505-5517, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32300856

RESUMO

ClpX and ClpP are involved in many important functions, including stress responses and energy metabolism, in microorganisms. However, the ClpX and ClpP of microbes used in industrial scale have rarely been studied. Industrial bacterial fermentation experiences a variety of stresses, and energy metabolism is extremely important for industrial bacteria. Thus, the role played by the ClpX and ClpP of industrial bacteria in fermentation should be investigated. Most microorganisms have a single clpP gene, while Corynebacterium crenatum AS 1.542 possesses two clpPs. Herein, the clpX, clpP1, and clpP2 of C. crenatum were cloned, and its fusion protein was expressed and characterized. We also constructed clpX deletion mutant and complementation strain. Results indicate that ClpX serves an important function in thermal, pH, and ethanol stresses. It is also involved in NADPH synthesis and glucose consumption during fermentation.


Assuntos
Corynebacterium/enzimologia , Endopeptidase Clp/metabolismo , Metabolismo Energético , Fermentação , Estresse Fisiológico , Clonagem Molecular , Corynebacterium/genética , Endopeptidase Clp/genética , Regulação Bacteriana da Expressão Gênica , Glucose/metabolismo , Microbiologia Industrial , Deleção de Sequência
12.
Appl Microbiol Biotechnol ; 104(12): 5303-5313, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32333052

RESUMO

Propionic acid (PA) is widely used in the food, agricultural, and pharmaceutical industries. Since the petrochemical PA is unsustainable, biological production of PA from renewable substrates is gaining attention. In this study, we engineered the strain Pseudomonas putida KT2440 to transform L-threonine to PA with only CO2 released as by-product. The cell factory was created by chromosomal incorporation of heterologous L-threonine deaminase, permease, and acyl-CoA thioesterase, deletion of branch pathways as well as overproduction of the endogenous branched-chain alpha-keto acid dehydrogenase complex. The final engineered strain could produce 399 mM PA from 400 mM L-threonine in a batch biotransformation process, with a molar yield of 99.8% under the optimized conditions in 48 h. The PA titer further reached to 50.3 g/L (679 mM) with a productivity of 0.6 g/L/h in a fed-batch conversion process. No obvious by-products, such as acetate and succinate, were detected in the broth, which would significantly facilitate downstream purification steps. Thus, this study offers an alternative route for biological production of PA.


Assuntos
Engenharia Metabólica/métodos , Propionatos/metabolismo , Pseudomonas putida/genética , Pseudomonas putida/metabolismo , Treonina/metabolismo , Biotransformação , Deleção de Genes , Microbiologia Industrial
13.
Appl Microbiol Biotechnol ; 103(21-22): 8839-8851, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31642949

RESUMO

D-p-hydroxyphenylglycine (D-HPG) functions as an intermediate and has important value in antibiotic industries. The high pollution and costs from chemical processes make biotechnological route for D-HPG highly desirable. Here, a whole-cell transformation process by D-hydantoinase(Hase) and D-carbamoylase(Case) was developed to produce D-HPG from DL-hydroxyphenylhydantoin(DL-HPH) in Escherichia coli. The artificially designed ribosome binding site with strong intensity significantly facilitated the protein expression of limiting step enzyme Case. Next, the cell wall permeability was improved by disturbing the peptidoglycan structure by overproduction of D,D-carboxypeptidases without obviously affecting cell growth, to increase the bioavailability of low soluble hydantoin substrate. By fine-tuning regulation of expression level of D,D-carboxypeptidase DacB, the final production yield of D-HPG increased to 100% with 140 mM DL-HPH substrate under the optimized transformation conditions. This is the first example to enhance bio-productivity of chemicals by cell wall engineering and creates a new vision on biotransformation of sparingly soluble substrates. Additionally, the newly demonstrated 'hydroxyl occupancy' phenomenon when Case reacts with hydroxyl substrates provides a referential information for the enzyme engineering in future.


Assuntos
Reatores Biológicos/microbiologia , Engenharia Celular/métodos , Parede Celular/genética , Escherichia coli/metabolismo , Glicina/análogos & derivados , Biossíntese de Proteínas/genética , Amidoidrolases/genética , Amidoidrolases/metabolismo , Carboxipeptidases/metabolismo , Parede Celular/metabolismo , Escherichia coli/genética , Engenharia Genética , Glicina/biossíntese , Permeabilidade , Biossíntese de Proteínas/fisiologia
14.
J Cardiovasc Pharmacol ; 67(2): 129-35, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26398164

RESUMO

It is well known that inflammation contributes to the development of coronary artery disease (CAD) and depressive symptoms. Previous studies have shown that long-term application of statin reduces the occurrence of depression in patients with CAD. However, the mechanism remains unclear. We hypothesized that inflammation contributes to depression in patients with CAD and statin function as an anti-inflammation therapy for those depressive patients. Patients with confirmed CAD hospitalized in the Department of Cardiology of Tongji Hospital in Shanghai, China, were enrolled. Depression was identified as none (ND), mild (MiD), moderate (MoD), or severe (SD) on the basis of scores of the patient health questionnaire with 9 items. Inflammatory factors in peripheral blood were measured using a chemiluminescence immunoassay and Bio-plex. Luciferase expression level was detected using the Dual-Luciferase Reporter Assay System for IL-1ß or NF-κB expression by transfection in human umbilical vein endothelial cells, and patient serum was added. Data obtained from 217 patients with CAD were analyzed. The IL-1ß level of CAD with SD was 14.70, which was significantly higher than that of CAD with ND 7.52, MiD 7.73, or MoD 8.63. Luciferase reporter gene analysis showed that IL-1ß or NF-κB expression level was upregulated by the serum of CAD and depression patients. After the addition of atorvastatin, IL-1ß or NF-κB luciferase reporter expression level decreased. It suggested that depression in patients with CAD is associated with inflammation. Statin may function as an anti-inflammation therapy for depression in patients with CAD by downregulation of IL-1ß.


Assuntos
Anti-Inflamatórios/uso terapêutico , Doença da Artéria Coronariana/tratamento farmacológico , Depressão/tratamento farmacológico , Regulação para Baixo/efeitos dos fármacos , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Interleucina-1beta/antagonistas & inibidores , Idoso , Idoso de 80 Anos ou mais , Anti-Inflamatórios/farmacologia , Doença da Artéria Coronariana/sangue , Doença da Artéria Coronariana/epidemiologia , Depressão/sangue , Depressão/epidemiologia , Regulação para Baixo/fisiologia , Feminino , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Interleucina-1beta/sangue , Masculino , Pessoa de Meia-Idade
15.
Appl Microbiol Biotechnol ; 100(17): 7471-8, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27020295

RESUMO

3-Phenyllactic acid (PLA) is an antimicrobial compound with broad-spectrum activity against bacteria and fungi that could be widely used in the food industry and livestock feeds. Notably, D-PLA exhibits higher antibacterial activity, which gains more attention than L-PLA. In this report, the D-lactate dehydrogenase DLDH744 from Sporolactobacillus inulinus CASD was engineered to increase the enzymatic activities toward phenylpyruvate by protein structure-guided modeling analysis. The phenylpyruvate molecule was first docked in the active center of DLDH744. The residues that might tightly pack around the benzene ring of phenylpyruvate were all selected for mutation. The single site mutant M307L showed the highest increased activity toward bulkier substrate phenylpyruvate than the wild type. By using the engineered D-lactate dehydrogenase M307L expressed in Escherichia coli strains, without coexpression of the cofactor regeneration system, 21.43 g/L D-PLA was produced from phenylpyruvate with a productivity of 1.58 g/L/h in the fed-batch biotransformation process, which ranked in the list as the highest production titer of D-PLA by D-lactate dehydrogenase. The enantiomeric excess value of produced D-PLA in the broth was higher than 99.7 %. Additionally, the structure-guided design of this enzyme will also provide referential information for further engineering other 2-hydroxyacid dehydrogenases, which are useful for a wide range of fine chemical synthesis.


Assuntos
Antibacterianos/síntese química , Bacillales/enzimologia , Biotransformação , Domínio Catalítico/genética , Lactato Desidrogenases/metabolismo , Lactatos/síntese química , Ácidos Fenilpirúvicos/metabolismo , Bacillales/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Lactato Desidrogenases/genética , Mutação , Estereoisomerismo
16.
Appl Microbiol Biotechnol ; 100(3): 1241-1251, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26454866

RESUMO

The pure stereoisomers of 1,2-propanediol (1,2-PDO) could be used as starting materials to synthesize high value-added specialty chemicals and chiral pharmaceutical products. As the stereoisomers of 1,2-PDO cannot be obtained by traditional chemical synthesis processes, biotechnological processes have gained increasing attention. However, to our knowledge, the production of S-1,2-PDO directly from glucose has not been previously reported. In this study, we demonstrate a novel artificial pathway to convert L-lactic acid to S-1,2-PDO and its integration into the genome of Escherichia coli strain BW25113∆poxB with synchronous deletion of genes responsible for branch metabolic pathways from glucose. L-lactate production was increased by replacing the native D-lactate dehydrogenase with the L-lactate dehydrogenase from Bacillus coagulans. The methylglyoxal bypass pathway was blocked to avoid synthesis of a racemic mixture of D- and L-lactate and prevent the accumulation of methylglyoxal, a toxic intermediate. To further improve the yield of S-1,2-PDO, a novel cofactor regeneration system was introduced by combining pyruvate decarboxylase and acetaldehyde-CoA dehydrogenase II to simultaneously regenerate NADH and the CoA donor of acetyl-CoA for the lactate conversion pathway. Finally, 13.7 mM S-1,2-PDO with >99 % enantiomeric purity was directly produced from glucose by disrupting the major carbon-competing pathways and strengthening the lactate transformation pathway. This study demonstrates the first attempt to synthesize S-1,2-PDO by direct fermentation of glucose.


Assuntos
Escherichia coli/metabolismo , Glucose/metabolismo , Engenharia Metabólica , Propilenoglicol/metabolismo , Escherichia coli/genética , Fermentação , Redes e Vias Metabólicas , Propilenoglicol/química , Estereoisomerismo
17.
Appl Environ Microbiol ; 81(18): 6294-301, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26150461

RESUMO

Hydroxy acid dehydrogenases, including l- and d-lactate dehydrogenases (L-LDH and D-LDH), are responsible for the stereospecific conversion of 2-keto acids to 2-hydroxyacids and extensively used in a wide range of biotechnological applications. A common feature of LDHs is their high specificity for NAD(+) as a cofactor. An LDH that could effectively use NADPH as a coenzyme could be an alternative enzymatic system for regeneration of the oxidized, phosphorylated cofactor. In this study, a d-lactate dehydrogenase from a Sporolactobacillus inulinus strain was found to use both NADH and NADPH with high efficiencies and with a preference for NADPH as its coenzyme, which is different from the coenzyme utilization of all previously reported LDHs. The biochemical properties of the D-LDH enzyme were determined by X-ray crystal structural characterization and in vivo and in vitro enzymatic activity analyses. The residue Asn(174) was demonstrated to be critical for NADPH utilization. Characterization of the biochemical properties of this enzyme will contribute to understanding of the catalytic mechanism and provide referential information for shifting the coenzyme utilization specificity of 2-hydroxyacid dehydrogenases.


Assuntos
Bacillales/enzimologia , Bacillales/metabolismo , Coenzimas/metabolismo , Lactato Desidrogenases/metabolismo , NADP/metabolismo , Sequência de Aminoácidos , Cristalografia por Raios X , Cinética , Lactato Desidrogenases/química , Modelos Moleculares , Dados de Sequência Molecular , NAD/metabolismo , Conformação Proteica , Homologia de Sequência de Aminoácidos
18.
Appl Environ Microbiol ; 80(23): 7134-41, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25217009

RESUMO

Bacillus coagulans 2-6 is an excellent producer of optically pure l-lactic acid. However, little is known about the mechanism of synthesis of the highly optically pure l-lactic acid produced by this strain. Three enzymes responsible for lactic acid production-NAD-dependent l-lactate dehydrogenase (l-nLDH; encoded by ldhL), NAD-dependent d-lactate dehydrogenase (d-nLDH; encoded by ldhD), and glycolate oxidase (GOX)-were systematically investigated in order to study the relationship between these enzymes and the optical purity of lactic acid. Lactobacillus delbrueckii subsp. bulgaricus DSM 20081 (a d-lactic acid producer) and Lactobacillus plantarum subsp. plantarum DSM 20174 (a dl-lactic acid producer) were also examined in this study as comparative strains, in addition to B. coagulans. The specific activities of key enzymes for lactic acid production in the three strains were characterized in vivo and in vitro, and the levels of transcription of the ldhL, ldhD, and GOX genes during fermentation were also analyzed. The catalytic activities of l-nLDH and d-nLDH were different in l-, d-, and dl-lactic acid producers. Only l-nLDH activity was detected in B. coagulans 2-6 under native conditions, and the level of transcription of ldhL in B. coagulans 2-6 was much higher than that of ldhD or the GOX gene at all growth phases. However, for the two Lactobacillus strains used in this study, ldhD transcription levels were higher than those of ldhL. The high catalytic efficiency of l-nLDH toward pyruvate and the high transcription ratios of ldhL to ldhD and ldhL to the GOX gene provide the key explanations for the high optical purity of l-lactic acid produced by B. coagulans 2-6.


Assuntos
Bacillus/enzimologia , Bacillus/metabolismo , Lactato Desidrogenases/metabolismo , Ácido Láctico/metabolismo , Bacillus/genética , Fermentação , Perfilação da Expressão Gênica , Cinética , Lactato Desidrogenases/genética , Lactobacillus delbrueckii/enzimologia , Lactobacillus delbrueckii/genética , Lactobacillus delbrueckii/metabolismo , Lactobacillus plantarum/enzimologia , Lactobacillus plantarum/genética , Lactobacillus plantarum/metabolismo
19.
Behav Sci (Basel) ; 14(4)2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38667088

RESUMO

An organizational ethical climate enhances the degree of collaboration and cohesion among employees and facilitates the development and interests of organizations. Such roles lead to organizational sustainable development and survival. Therefore, the importance of ethical climate in organizations is becoming increasingly apparent. In this background, this study aims to explore whether an organizational ethical climate can improve whistleblowing behavior and the mediating role of organizational identification in promoting whistleblowing behavior. Most previous studies have only focused on the mediating or moderating role of the model. This study expands the research field, adds the dual moderation of person-organization value congruence and leader ethical behavior, and verifies two moderated mediation models. Overall, the purpose of this study is to determine the behavior of employees under the influence of an organizational ethical climate and, on this basis, propose suggestions for strengthening organizational ethical climate, expanding the scope of research on organizational climate and providing a theoretical basis for related research. In order to achieve the research goals, the data were collected from 344 Chinese SMEs for empirical analysis. The results showed that an organizational ethical climate has no direct impact on whistleblowing behavior but could have a positive effect on whistleblowing formation through the mediating variable of organizational identification. In addition, person-organization value congruence and leader ethical behavior significantly moderated the mediating role of organizational identification between organizational ethical climate and whistleblowing behavior. Finally, the directions that can contribute to future research were suggested.

20.
Inquiry ; 61: 469580241249425, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38727154

RESUMO

Quality improvement is an international priority, and quality education and training are important parts of hospital quality management. The aim of this study was to understand the knowledge, attitudes and practices (KAP) and its influencing factors related to quality training in medical staff. A questionnaire survey was conducted by convenience sampling to assess the KAP of quality training in Taizhou Enze Medical Center. Principal component analysis was used to extract factors from the questionnaire. Descriptive statistics (frequency, median, mean), Kendall grade correlation analysis, and Mann-Whitney U tests were used to analyze the data. A total of 205 staff members participated in the questionnaire survey. For the 5 factors of the KAP scale, the highest score was factor F4, recognition and support for quality training (mean = 90.55, median = 100), followed by factor F3, perceived benefits (mean = 84.46, median = 85.65). Relatively lower scores were found for factor F2, quality knowledge learning and mastery (mean = 63.09, median = 63.89), and F5, quality management practices and sharing (mean = 82.07, median = 75.00). There was a correlation between the 5 factors. The scores of F2 (quality knowledge learning and mastery) for staff with senior professional titles were higher than those for staff with intermediate professional titles or below. The score of F3 (perceived benefits of quality training) in medical technicians and nurses was higher than in doctors and administrative personnel. Our findings showed that the respondents' attitude toward quality training was positive, but their knowledge mastery and practice behaviors should be further improved. Occupational category and professional title were the influencing factors of the quality training KAP. Therefore, hospital should conduct quality management training at a wider scope according to the competency requirements of different groups, and further optimize the improvement and innovation system.


Assuntos
Conhecimentos, Atitudes e Prática em Saúde , Hospitais Gerais , Centros de Atenção Terciária , Humanos , Estudos Transversais , Masculino , Feminino , Adulto , Inquéritos e Questionários , Melhoria de Qualidade , Pessoa de Meia-Idade , Atitude do Pessoal de Saúde , Corpo Clínico Hospitalar , China
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