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1.
Rev Soc Bras Med Trop ; 52: e20190237, 2019 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-31508785

RESUMO

INTRODUCTION: The increased use of colistin against infections caused by Acinetobacter baumannii and Pseudomonas aeruginosa has resulted in colistin resistance. The purpose of this study was to detect plasmid-mediated mcr-1 gene in colistin-resistant A. baumannii and P. aeruginosa isolates. METHODS: A total of 146 clinical isolates of A. baumannii (n = 62) and P. aeruginosa (n = 84) were collected from the four largest tertiary care hospitals in Peshawar, Pakistan. All bacterial isolates were phenotypically screened for multidrug resistance using the Kirby-Baur disc diffusion method. The minimum inhibitory concentration (MIC) of colistin in all isolates was phenotypically performed using dilution methods. mcr-1 gene was detected through polymerase chain reaction and the nucleotide sequence of amplicon was determined using Sanger sequencing. RESULTS: Approximately 96.7% A. baumannii and 83.3% P. aeruginosa isolates were resistant to multiple antibiotics. Colistin resistance was found in 9.6% (6/62) of A. baumannii and 11.9% (10/84) of P. aeruginosa isolates. Among 16 colistin resistant isolates, the mcr-1 gene was detected in one A. baumannii (1.61% of total isolates; 16.6% of colistin resistant isolates) and one P. aeruginosa strain (1.19% of total isolates; 10% of colistin resistant isolates). Nucleotide BLAST showed 98-99% sequence similarity to sequences of the mcr-1 gene in GenBank. CONCLUSIONS: Our study reports, for the first time, the emergence of plasmid-mediated mcr-1-encoded colistin resistance in multidrug resistant strains of A. baumannii and P. aeruginosa. Further large scales studies are recommended to investigate the prevalence of this mode of resistance in these highly pathogenic bacteria.


Assuntos
Infecções por Acinetobacter/microbiologia , Acinetobacter baumannii/genética , Proteínas de Bactérias/genética , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/genética , Acinetobacter baumannii/efeitos dos fármacos , Farmacorresistência Bacteriana , Humanos , Testes de Sensibilidade Microbiana , Paquistão , Plasmídeos/genética , Pseudomonas aeruginosa/efeitos dos fármacos
2.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(7): 589-594, 2019 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-31537242

RESUMO

Objective To construct and identify Bifidobacterium bifidum-vectored outer membrane protein F-I[rBb(pGEX-OprF-I)] vaccine of Pseudomonas aeruginosa and observe its protection against Pseudomonas aeruginosa infection in mice. Methods OprF and OprI genes were amplified by PCR, then the OprF-I fusion gene obtained by gene SOEing was digested and ligated into the vector pGEX-1λT to construct the recombinant plasmid pGEX-OprF-I. The plasmid was transformed into Bifidobacterium bifidum (Bb) by electroporation, and the rBb(pGEX-OprF-I) vaccine was constructed and identified by double enzyme digestion and PCR. Expression products of the vaccine induced by IPTG were analyzed and identified by SDS-PAGE and Western blot analysis. Twenty-one BALB/c mice were randomly divided into rBb(pGEX-OprF-I) vaccine group, Bb-pGEX-1λT empty vector group and Bb control group. The 5×108 CFUs vaccine was intragastrically administered for 3 consecutive days per week for 3 weeks. All mice were challenged intranasally with 5×107 CFUs of PA01 strain at the 4th week after the first immunization. At the 2nd week after the challenge, all mice were sacrificed to count the lung bacteria loads. IgG levels in sera from the mice before immunization, 4th week after the first immunization and 2nd week after the challenge were detected by routine ELISA. Results A total of 1289 bp OprF-I fusion gene was amplified by PCR. Double enzyme digestion and PCR identification confirmed that the gene was ligated into pGEX-1λT and transformed into Bb, and the rBb(pGEX-OprF-I) vaccine was successfully constructed. SDS-PAGE showed that the fusion protein with a relative molecular mass (Mr) of about 68 000 could be expressed by IPTG-induced vaccine. Western blot analysis indicated that the protein could be specifically recognized by the sera of Pseudomonas aeruginosa-infected mice. The number of bacteria colonies in the lung of the mice immunized with rBb(pGEX-OprF-I) vaccine was significantly lower than that of the control group. The IgG levels in the sera of the immunized mice increased successively at 4th week after the first immunization and 2nd week after the challenge, and higher than that in the other control groups at the same time point. Conclusion The rBb(pGEX-OprF-I) vaccine has been successfully constructed, and it may take a certain protective effect on the mice against Pseudomonas aeruginosa infection.


Assuntos
Proteínas de Bactérias/imunologia , Bifidobacterium bifidum , Lipoproteínas/imunologia , Infecções por Pseudomonas/prevenção & controle , Vacinas contra Pseudomonas/imunologia , Animais , Camundongos , Camundongos Endogâmicos BALB C , Pseudomonas aeruginosa , Distribuição Aleatória , Proteínas Recombinantes de Fusão/imunologia
3.
Cochrane Database Syst Rev ; 9: CD002009, 2019 09 04.
Artigo em Inglês | MEDLINE | ID: mdl-31483853

RESUMO

BACKGROUND: People with cystic fibrosis, who are chronically colonised with the organism Pseudomonas aeruginosa, often require multiple courses of intravenous aminoglycoside antibiotics for the management of pulmonary exacerbations. The properties of aminoglycosides suggest that they could be given in higher doses less often. This is an update of a previously published review. OBJECTIVES: To assess the effectiveness and safety of once-daily versus multiple-daily dosing of intravenous aminoglycoside antibiotics for the management of pulmonary exacerbations in cystic fibrosis. SEARCH METHODS: We searched the Cystic Fibrosis Specialist Register held at the Cochrane Cystic Fibrosis and Genetic Disorders Group's editorial base, comprising references identified from comprehensive electronic database searches, handsearching relevant journals and handsearching abstract books of conference proceedings.Date of the most recent search: 31 January 2019.We also searched online trial registries. Date of latest search: 25 February 2019. SELECTION CRITERIA: All randomised controlled trials, whether published or unpublished, in which once-daily dosing of aminoglycosides has been compared with multiple-daily dosing in terms of efficacy or toxicity or both, in people with cystic fibrosis. DATA COLLECTION AND ANALYSIS: The two authors independently selected the studies to be included in the review and assessed the risk of bias of each study; authors also assessed the quality of the evidence using the GRADE criteria. Data were independently extracted by each author. Authors of the included studies were contacted for further information. As yet unpublished data were obtained for one of the included studies. MAIN RESULTS: We identified 15 studies for possible inclusion in the review. Five studies reporting results from a total of 354 participants (aged 5 to 50 years) were included in this review. All studies compared once-daily dosing with thrice-daily dosing. One cross-over trial had 26 participants who received the first-arm treatment but only 15 received the second arm. One study had a low risk of bias for all criteria assessed; the remaining included studies had a high risk of bias from blinding, but for other criteria were judged to have either an unclear or a low risk of bias.There was little or no difference between treatment groups in: forced expiratory volume in one second, mean difference (MD) 0.33 (95% confidence interval (CI) -2.81 to 3.48, moderate-quality evidence); forced vital capacity, MD 0.29 (95% CI -6.58 to 7.16, low-quality evidence); % weight for height, MD -0.82 (95% CI -3.77 to 2.13, low-quality evidence); body mass index, MD 0.00 (95% CI -0.42 to 0.42, low-quality evidence); or in the incidence of ototoxicity, relative risk 0.56 (95% CI 0.04 to 7.96, moderate-quality evidence). Once-daily treatment in children probably improved the percentage change in creatinine, MD -8.20 (95% CI -15.32 to -1.08, moderate-quality evidence), but showed no difference in adults, MD 3.25 (95% CI -1.82 to 8.33, moderate-quality evidence). The included trials did not report antibiotic resistance patterns or quality of life. AUTHORS' CONCLUSIONS: Once- and three-times daily aminoglycoside antibiotics appear to be equally effective in the treatment of pulmonary exacerbations of cystic fibrosis. There is evidence of less nephrotoxicity in children.


Assuntos
Aminoglicosídeos/uso terapêutico , Antibacterianos/uso terapêutico , Fibrose Cística/tratamento farmacológico , Pneumopatias , Aminoglicosídeos/administração & dosagem , Antibacterianos/administração & dosagem , Fibrose Cística/complicações , Esquema de Medicação , Quimioterapia Combinada/métodos , Volume Expiratório Forçado/efeitos dos fármacos , Humanos , Injeções Intravenosas , Pneumopatias/tratamento farmacológico , Pneumopatias/microbiologia , Infecções por Pseudomonas/tratamento farmacológico , Pseudomonas aeruginosa , Ensaios Clínicos Controlados Aleatórios como Assunto , Capacidade Vital/efeitos dos fármacos
4.
Klin Lab Diagn ; 64(8): 497-502, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31479607

RESUMO

The growing prevalence of metallo-ß-lactamase (MBL)-producing Pseudomonas aeruginosa in nosocomial pathogen populations has been attributed to their clonal spread, and/or horizontal transfer of MBL determinants in mobile genetic elements, including integrons. To characterize the genetic background of the beta-lactamase VIM-2 encoding gene in the population of carbapenemresistant (Carba-R) P. aeruginosa clinical isolates.The detection of class 1 integrons was performed by PCR. Typing of the class 1 integrons containing the blaVIM gene cassette was performed by the PCR-restriction fragment length polymorphism (RFLP) approach followed by sequencing of variable regions of class 1 integrons. Five types of the blaVIM-2-carrying integrons were identified: ST654-isolates accounting for more than 50% of the Carba-R population harbored In56; ST235-isolates contained In559 (26% Carba-R isolates); ST111-isolates (19% Carba-R isolates) were characterized by carrying In59-like integron; two ST235-isolates harbored In59 and In249 each. Except In56, carrying the only blaVIM-2-gene cassette, all other identified integron types harbored the genes of resistance to trimethoprim and/or aminoglycosides. No new types of integrons were identified in the P. aeruginosa clinical isolates. The observed correlation of the integron type with specific STs indicates a clonal dissemination of significant resistance determinant producers - ST111, ST654 and ST235 epidemic lines. The features of the integron variable regions can be used for the epidemiological characterization of clinical P. aeruginosa isolates.


Assuntos
Carbapenêmicos/farmacologia , Farmacorresistência Bacteriana , Integrons , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/genética , beta-Lactamases/genética , Antibacterianos/farmacologia , Humanos , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/efeitos dos fármacos
5.
Klin Lab Diagn ; 64(8): 503-506, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31479608

RESUMO

The results of the bacteriological catheter investigation on the analyzer with the technology of laser light scattering and using D. Maki culture technique coincided in 91.8% of cases. Catheter-associated infections are proven in 8 cases. The following obtained from blood and intravascular catheter in 5 patients: Staphylococcus epidermidis (n=1), Staphylococcus haemolyticus (n=1), Escherichia coli (n=1), Staphylococcus aureus (n=2); Staphylococcus epidermidis (n=1) was detected in 1 child from ventricular shunt and cerebrospinal fluid and Escherichia coli lactose-negative + Staphylococcus haemolyticus were detected in 1 child from ventricular shunt, whily only Escherichia coli lactose-negative was detected from cerebrospinal fluid; Pseudomonas aeruginosa (n=1) was foud out in 1 patient from the urinary catheter and urine. Clinical significance of the isolated microorganisms from the catheter must be assessed in each particular case taking into account its quantity and type of the isolated microorganism.


Assuntos
Infecções Bacterianas/diagnóstico , Infecções Relacionadas a Cateter/diagnóstico , Lasers , Criança , Escherichia coli , Humanos , Pseudomonas aeruginosa , Staphylococcus aureus , Staphylococcus epidermidis , Staphylococcus haemolyticus
6.
J Enzyme Inhib Med Chem ; 34(1): 1414-1425, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31401901

RESUMO

The emergence of drug-resistant pathogenic bacteria is occurring due to the global overuse and misuse of ß-lactam antibiotics. Infections caused by some bacteria which secrete metallo-ß-lactamases (enzymes that inactivate ß-lactam antibiotics) are increasingly prevalent and have become a major worldwide threat to human health. These bacteria are resistant to ß-lactam antibiotics and MBL-inhibitor/ß-lactam antibiotic combination therapy can be a strategy to overcome this problem. So far, no clinically available inhibitors of metallo-ß-lactamases (MBLs) have been reported. In this study, L-benzyl tyrosine thiol carboxylic acid analogues (2a-2k) were synthesized after the study of computational simulation by adding of methyl, chloro, bromo and nitro groups to the benzyl ring for investigation of SAR analysis. Although the synthesized molecules 2a-k shows the potent inhibitory effects against metallo-ß-lactamase (IMP-1) with the range of Kic values of 1.04-4.77 µM, they are not as potent as the candidate inhibitor.


Assuntos
Ácidos Carboxílicos/síntese química , Ácidos Carboxílicos/farmacologia , Compostos de Sulfidrila/química , Tirosina/química , Inibidores de beta-Lactamases/síntese química , Inibidores de beta-Lactamases/farmacologia , beta-Lactamases/efeitos dos fármacos , Antibacterianos/síntese química , Antibacterianos/química , Antibacterianos/farmacologia , Espectroscopia de Ressonância Magnética Nuclear de Carbono-13 , Ácidos Carboxílicos/química , Espectrometria de Massas , Testes de Sensibilidade Microbiana , Espectroscopia de Prótons por Ressonância Magnética , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/enzimologia , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/enzimologia , Relação Estrutura-Atividade , Inibidores de beta-Lactamases/química
7.
World J Microbiol Biotechnol ; 35(9): 137, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31432268

RESUMO

The presence of very high concentrations of organic pollutants, phenols, tannins and heavy metals mainly chromium in wastewater discharged from leather industries, tags it as one of the most polluting industries. The phenolic syntans discharged from tanning units have an adverse effect on living organisms and cause serious environmental pollution, thereby making it very imperative to remove it. Among various treatment methods available for removal of phenols, biodegradation is environment friendly. The present study aims at the remediation of phenolic syntan used in the leather industry employing individual as well as co-culture of Bacillus cereus and Pseudomonas aeruginosa at varying syntan concentration in the medium. Parameters such as chemical oxygen demand (COD), total organic carbon (TOC), total phenol content (TPC) and Fourier Transform Infrared Spectroscopy (FTIR) indicating biodegradation were analyzed. Promising results were observed with P. aeruginosa, which exhibited a reduction in TPC by 62-72% in all the concentrations of syntan tested just within 12 h of inoculation, whereas about 67 and 83% reduction in COD and TOC respectively was observed for 2000 ppm concentration at the end of 5 days. B. cereus also demonstrated very good reduction in the above parameters however; percentage was less as compared to P. aeruginosa. In the case of co-culture, the TPC reduction was higher than B. cereus but lesser than P. aeruginosa. The percentage reduction in TOC and COD was highest for 500 ppm which eventually decreased for subsequent concentrations.


Assuntos
Bacillus cereus/metabolismo , Resíduos Industriais , Fenóis/metabolismo , Pseudomonas aeruginosa/metabolismo , Poluentes Químicos da Água/metabolismo , Bacillus cereus/crescimento & desenvolvimento , Análise da Demanda Biológica de Oxigênio , Biotransformação , Carbono/análise , Técnicas de Cocultura , Fenóis/análise , Pseudomonas aeruginosa/crescimento & desenvolvimento , Espectroscopia de Infravermelho com Transformada de Fourier , Água/química
9.
Mem Inst Oswaldo Cruz ; 114: e190105, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31389522

RESUMO

BACKGROUND: Healthcare-associated infections caused by bacteria such as Pseudomonas aeruginosa are a major public health problem worldwide. Gene regulatory networks (GRN) computationally represent interactions among regulatory genes and their targets. They are an important approach to help understand bacterial behaviour and to provide novel ways of overcoming scientific challenges, including the identification of potential therapeutic targets and the development of new drugs. OBJECTIVES: The goal of this study was to reconstruct the multidrug-resistant (MDR) P. aeruginosa GRN and to analyse its topological properties. METHODS: The methodology used in this study was based on gene orthology inference using the reciprocal best hit method. We used the genome of P. aeruginosa CCBH4851 as the basis of the reconstruction process. This MDR strain is representative of the sequence type 277, which was involved in an endemic outbreak in Brazil. FINDINGS: We obtained a network with a larger number of regulatory genes, target genes and interactions as compared to the previously reported network. Topological analysis results are in accordance with the complex network representation of biological processes. MAIN CONCLUSIONS: The properties of the network were consistent with the biological features of P. aeruginosa. To the best of our knowledge, the P. aeruginosa GRN presented here is the most complete version available to date.


Assuntos
Farmacorresistência Bacteriana Múltipla/genética , Redes Reguladoras de Genes , Pseudomonas aeruginosa/genética , Genoma Bacteriano , Família Multigênica , Infecções por Pseudomonas/genética , Valores de Referência
11.
Adv Exp Med Biol ; 1145: 1-8, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31364067

RESUMO

Antibiotic resistance has become the most significant threat to human health across the globe. Polymyxins are often used as the only available therapeutic option against Gram-negative 'superbugs', namely Acinetobacter baumannii, Pseudomonas aeruginosa and Klebsiella pneumoniae. The limited pharmacological and clinical knowledge on the polymyxins in the old literature substantially limited optimizing their clinical use. The current chapter provides a general introduction to this first-ever polymyxin book which comprehensively reviews the significant progress over the last two decades in the chemistry, microbiology, pharmacology, clinical use and drug discovery of polymyxins. In particular, recent pharmacological results have led to the first scientifically-based dosing recommendations and facilitated the discovery of new-generation polymyxins. Future challenges in polymyxin research are highlighted, aiming at improving the clinical utility of this last-line defence.


Assuntos
Antibacterianos/farmacologia , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Polimixinas/farmacologia , Acinetobacter baumannii/efeitos dos fármacos , Humanos , Pseudomonas aeruginosa/efeitos dos fármacos
12.
Adv Exp Med Biol ; 1145: 9-13, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31364068

RESUMO

Antibiotic resistance has presented a major health challenge in the world and many isolates of Enterobacteriaceae, Acinetobacter baumannii and Pseudomonas aeruginosa become resistant to almost all current antibiotics. This chapter provides an overview on the mechanisms of antibiotic resistance in these Gram-negative pathogens and outlines the formidable problem of the genetics of bacterial resistance. Prevalent multidrug-resistance in Gram-negative bacteria underscores the need for optimizing the clinical use of the last-line polymyxins.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Polimixinas/farmacologia , Acinetobacter baumannii/efeitos dos fármacos , Bactérias Gram-Negativas/efeitos dos fármacos , Humanos , Pseudomonas aeruginosa/efeitos dos fármacos
13.
Adv Exp Med Biol ; 1145: 105-116, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31364074

RESUMO

In the last decade, considerable advancements have been made to identify the pharmacokinetic/pharmacodynamic (PK/PD) index that defines the antimicrobial activity of polymyxins. Dose-fractionation studies performed in hollow-fiber models found that altering the dosing schedule had little impact on the killing or suppression of resistance emergence, alluding to AUC/MIC as the pharmacodynamic index that best describes polymyxin's activity. For in vivo efficacy, the PK/PD index that was the most predictive of the antibacterial effect of colistin against P. aeruginosa and A. baumannii was ƒAUC/MIC.


Assuntos
Antibacterianos/farmacologia , Antibacterianos/farmacocinética , Polimixinas/farmacologia , Polimixinas/farmacocinética , Acinetobacter baumannii/efeitos dos fármacos , Animais , Colistina/farmacocinética , Colistina/farmacologia , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/efeitos dos fármacos
14.
Ying Yong Sheng Tai Xue Bao ; 30(8): 2767-2774, 2019 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-31418202

RESUMO

To reveal the physiological effects of rice alleviated by cadmium-tolerant Pseudomonas aeruginosa under cadmium stress condition, the influences of bacterial strian on the root vigor and leaf physiological characteristics were analyzed under a set of hydroponic experiments involving adding bacteria suspension, empty carrier, microbial inoculum with 20 µmol·L-1 Cd. Cadmium-free treatment as control. The results showed that the root vigor was significantly inhibited, leaf photosynthetic rate decreased, and the contents of soluble protein, flavonoid and total phenols in rice leaves were reduced, while the contents of malondialdehyde (MDA) and superoxide anion(O2-·) increased significantly under cadmium stress condition. Compared with cadmium treatment, root vigors of rice were increased by 36.1%-42.5% and 49.4%-53.0% respectively in bacteria suspension and microbial inoculum additions, net photosynthetic rates in leaves were increased by 118.5%-147.1% and 137.6%-156.9%, and the contents of soluble protein were increased by 37.0%-49.3% and 37.7%-72.6%, respectively. For the bacteria suspension treatment, the activities of SOD, POD and CAT in leaves were increased by 25.8%-36.6%, 40.9%-55.9%, 24.0%-29.2%, and the activities of SOD, POD and CAT in leaves under microbial inoculum treatment were increased by 36.9%-42.6%, 82.7%-92.6% and 43.3%-52.2%, respectively, with the stimulative effects on antioxidation enzymes in rice leaves being higher than those of bacteria suspension. Compared with cadmium treatment, the contents of MDA and O2-· in rice leaves were reduced by 44.8%-54.7%, 29.4%-41.9% and 9.9%-10.2%, 3.0%-7.1% in microbial inoculum and bacteria suspension treatments, respectively. In contrast, the contents of flavonoids and total phenols were increased by 125.4%-135.7%, 100.8%-119.4% and 139.4%-146.7%, 115.0%-134.7%, respectively. In summary, P. aeruginosa and the microbial inoculum could promote rice seedling growth by improving root vigor and photosynthetic rate, as well as the contents of flavonoids and total phenols, which led to the fact that P. aeruginosa could significantly alleviate the stress of cadmium on rice.


Assuntos
Cádmio/toxicidade , Oryza/fisiologia , Folhas de Planta/fisiologia , Raízes de Plantas/fisiologia , Pseudomonas aeruginosa/fisiologia , Poluentes do Solo/toxicidade , Oryza/microbiologia , Folhas de Planta/microbiologia , Raízes de Plantas/microbiologia , Plântula , Estresse Fisiológico
15.
J Agric Food Chem ; 67(35): 9749-9756, 2019 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-31415718

RESUMO

Bovine lactoferrin N-lobe plays an important key in the nonimmunological defense system. In this work, the most suitable promoter Pveg was selected and the fragment coding bovine lactoferrin N-lobe was optimized according to codon bias of Bacillus. The recombinant plasmid pMA0911-Pveg-mBLF-N was introduced into Baicillus subtilis 168 to create B. subtilis/pMA0911-Pveg-mBLF-N. The bovine lactoferrin N-lobe was highly expressed at 28 °C for 15 h. Its purified protein was obtained with 16.5 mg/L and a purity of 93.6% using ammonium sulfate precipitation, Ni-NTA, and molecular exclusion. About 200 ng/mL purified bovine lactoferrin N-lobe completely inhibited cell-growth of Escherichia coli JM109 (DE3), 70.3% of Pseudomonas aeruginosa CGMCC 1.6740, and 41.5% of Staphylococcus aureus CGMCC 1.282. To our knowledge, this is the first report about active expression, purification, and characterization of bovine lactoferrin N-lobe in safe bacterium B. subtilis, which opens an available application way in the biomedical and food industries.


Assuntos
Antibacterianos/farmacologia , Bacillus subtilis/genética , Códon/genética , Lactoferrina/genética , Regiões Promotoras Genéticas , Animais , Antibacterianos/metabolismo , Bacillus subtilis/metabolismo , Clonagem Molecular , Códon/metabolismo , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Expressão Gênica , Engenharia Genética , Lactoferrina/metabolismo , Lactoferrina/farmacologia , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/crescimento & desenvolvimento , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento
16.
J Agric Food Chem ; 67(31): 8581-8589, 2019 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-31321975

RESUMO

Intermediates in aromatic amino acid biosynthesis can serve as substrates for the synthesis of bioactive compounds. In this study we used two intermediates in the shikimate pathway of Escherichia coli, chorismate and anthranilate, to synthesize three bioactive compounds: 4-hydroxycoumarin (4-HC), 2,4-dihydroxyquinoline (DHQ), and 4-hydroxy-1-methyl-2(1H)-quinolone (NMQ). We introduced genes for the synthesis of salicylic acid from chorismate to supply the substrate for 4-HC and the gene encoding N-methyltransferase for the synthesis of N-methylanthranilate from anthranilate. Polyketide synthases and coenzyme (Co)A ligases were tested to determine the optimal combination of genes for the synthesis of each compound. We also tested several constructs and identified the best one for increasing levels of endogenous substrates for chorismate, anthranilate, and malonyl-CoA. With the use of these strategies, 255.4 mg/L 4-HC, 753.7 mg/L DHQ, and 17.5 mg/L NMQ were synthesized. This work provides a basis for the synthesis of diverse coumarin and quinoline derivatives with potential medical applications.


Assuntos
4-Hidroxicumarinas/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Engenharia Metabólica , Policetídeo Sintases/genética , Quinolinas/metabolismo , 4-Hidroxicumarinas/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Ácido Corísmico/metabolismo , Coenzima A Ligases/genética , Coenzima A Ligases/metabolismo , Photorhabdus/enzimologia , Photorhabdus/genética , Policetídeo Sintases/metabolismo , Pseudomonas aeruginosa/enzimologia , Pseudomonas aeruginosa/genética , Quinolinas/química , ortoaminobenzoatos/metabolismo
17.
Microbiol Res ; 226: 19-26, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31284940

RESUMO

Pseudomonas aeruginosa is one of the most common pathogens associated with nosocomial infections and a great concern to immunocompromised individuals especially in the cases of cystic fibrosis, AIDS and burn wounds. The pathogenicity of P. aeruginosa is largely directed by the quorum sensing (QS) system. Hence, QS may be considered an important therapeutic target to combat P. aeruginosa infections. The anti-quorum sensing and anti-biofilm efficacy of aromatic aldehyde, 5-hydroxymethylfurfural (5-HMF) against P. aeruginosa PAO1 were assessed. At the sub-inhibitory concentration, 5-HMF suppressed the production of QS-controlled virulence phenotypes and biofilm formation in P. aeruginosa. It was also able to significantly enhance the survival rate of C. elegans infected with P. aeruginosa. The in silico studies revealed that 5-HMF could serve as a competitive inhibitor for the auto-inducer molecules as it exhibited a strong affinity for the regulatory proteins of the QS-circuits i.e. LasR and RhlR. In addition, a significant down-regulation in the expression of QS-related genes was observed suggesting the ability of 5-HMF in mitigating the pathogenicity of P. aeruginosa.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Furaldeído/análogos & derivados , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/patogenicidade , Percepção de Quorum/efeitos dos fármacos , Animais , Proteínas de Bactérias , Caenorhabditis elegans , Simulação por Computador , Modelos Animais de Doenças , Furaldeído/farmacologia , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Simulação de Acoplamento Molecular , Percepção de Quorum/genética , Taxa de Sobrevida , Transativadores , Virulência/efeitos dos fármacos , Fatores de Virulência
18.
J Nanobiotechnology ; 17(1): 81, 2019 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-31286976

RESUMO

BACKGROUND: Magnetic nanoparticles (MNPs) are characterized by unique physicochemical and biological properties that allow their employment as highly biocompatible drug carriers. Gelsolin (GSN) is a multifunctional actin-binding protein involved in cytoskeleton remodeling and free circulating actin sequestering. It was reported that a gelsolin derived phosphoinositide binding domain GSN 160-169, (PBP10 peptide) coupled with rhodamine B, exerts strong bactericidal activity. RESULTS: In this study, we synthesized a new antibacterial and antifungal nanosystem composed of MNPs and a PBP10 peptide attached to the surface. The physicochemical properties of these nanosystems were analyzed by spectroscopy, calorimetry, electron microscopy, and X-ray studies. Using luminescence based techniques and a standard killing assay against representative strains of Gram-positive (Staphylococcus aureus MRSA Xen 30) and Gram-negative (Pseudomonas aeruginosa Xen 5) bacteria and against fungal cells (Candida spp.) we demonstrated that magnetic nanoparticles significantly enhance the effect of PBP10 peptides through a membrane-based mode of action, involving attachment and interaction with cell wall components, disruption of microbial membrane and increased uptake of peptide. Our results also indicate that treatment of both planktonic and biofilm forms of pathogens by PBP10-based nanosystems is more effective than therapy with either of these agents alone. CONCLUSIONS: The results show that magnetic nanoparticles enhance the antimicrobial activity of the phosphoinositide-binding domain of gelsolin, modulate its mode of action and strengthen the idea of its employment for developing the new treatment methods of infections.


Assuntos
Antibacterianos/química , Antifúngicos/química , Gelsolina/química , Nanopartículas de Magnetita/química , Fragmentos de Peptídeos/química , Biofilmes , Candida/efeitos dos fármacos , Membrana Celular/metabolismo , Ouro/química , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Nanoconchas/química , Plâncton , Pseudomonas aeruginosa/efeitos dos fármacos , Rodaminas/química
19.
J Water Health ; 17(4): 532-539, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31313992

RESUMO

Pseudomonas aeruginosa is an environmental bacterium, ubiquitous in aquatic habitats and water distribution systems, including dental unit waterlines (DUWLs). We investigated the prevalence of P. aeruginosa in DUWLs from private dental settings. We also analyzed the relationship between P. aeruginosa contamination and the presence of Legionella spp. and total viable count (TVC) in order to suggest a simple and inexpensive protocol to test the quality of water from DUWLs. We detected and quantified P. aeruginosa both by culture and by a PMA (propidium monoazide)-qPCR method. Overall, we detected P. aeruginosa in 17 samples using the PMA-qPCR and in 11 samples using the culture. All culture-positive samples were positive with the PMA-qPCR too, with an agreement between the two methods of 93% and a Cohen's kappa coefficient of κ = 0.747 (good concordance). Comparing results with results of our previous study, we noted that (a) P. aeruginosa was isolated only from DUWLs with high TVC and (b) five out of six Legionella-positive samples were negative for Pseudomonas spp. Our final suggestion is that the cleanliness of DUWLs should be assessed by TVC because it is a good indicator of the presence of pathogens such as Legionella spp. and P. aeruginosa.


Assuntos
Equipamentos Odontológicos , Pseudomonas aeruginosa/crescimento & desenvolvimento , Microbiologia da Água , Bactérias , Contaminação de Equipamentos
20.
J Agric Food Chem ; 67(29): 8191-8196, 2019 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-31282662

RESUMO

Conversion of free fatty acids into monoacylglycerol gives rise to new structural properties, particularly amphipathic property. Therefore, monoacylglycerols are widely used in pharmaceutical and food industries and are also reported to facilitate better absorption into the human body. A functional fatty acid when transformed into a monoacylglycerol will possibly conserve both the original functionality and amphipathic property. The compound 7,10-dihydroxy-8(E)-octadecenoic acid (DOD) was generated from oleic acid by Pseudomonas aeruginosa PR3 and was known to contain antimicrobial activities against a broad range of food-borne and plant pathogenic bacteria. Here, we attempted to convert DOD into its monoacylglycerol form using lipase for producing an amphipathic antibacterial agent. Consequently, the monoacylglycerol of DOD (DOD-MAG) was successfully produced by coincubating DOD, glycerol, and lipase at 30 °C. The maximum conversion yield reached 70% after 12 h of incubation. Antibacterial activity of DOD-MAG was enhanced by 8 times from the original activity of DOD against food-borne bacteria.


Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Monoglicerídeos/química , Ácidos Oleicos/química , Ácidos Oleicos/farmacologia , Pseudomonas aeruginosa/química , Antibacterianos/metabolismo , Microbiologia de Alimentos , Ácidos Oleicos/metabolismo , Pseudomonas aeruginosa/metabolismo , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento
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