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1.
J Antibiot (Tokyo) ; 77(7): 471-474, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38664572

RESUMO

Benastatin K (1), a new chlorinated benastatin derivative, was isolated from the culture broth of the actinomycete Streptomyces sp. HGTA384. The structure of 1 was determined on the basis of spectroscopic analysis, including 1D and 2D NMR, as well as HRESI-MS, UV and IR, and comparison with data reported in the literature. Compound 1 and benastatins A and B exhibited inhibitory activity against Micrococcus luteus (MIC 7.8, 31.3, and 3.9 µM, respectively), and IgE-mediated ß-hexosaminidase release in RBL-2H3 cells with IC50 values of 42, 79, and 19 µM, respectively.


Assuntos
Antibacterianos , Espectroscopia de Ressonância Magnética , Testes de Sensibilidade Microbiana , Micrococcus luteus , Streptomyces , Streptomyces/química , Streptomyces/metabolismo , Micrococcus luteus/efeitos dos fármacos , Antibacterianos/farmacologia , Antibacterianos/isolamento & purificação , Antibacterianos/química , Animais , Ratos , beta-N-Acetil-Hexosaminidases/antagonistas & inibidores , beta-N-Acetil-Hexosaminidases/metabolismo , Concentração Inibidora 50 , Linhagem Celular Tumoral , Estrutura Molecular
2.
Braz J Microbiol ; 55(2): 1139-1150, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38378880

RESUMO

In recent years, some microorganisms have shown resistance to conventional treatments. Considering this increase in resistant pathogens, treatment alternatives are needed to promote greater treatment efficiency. In this sense, antimicrobial photodynamic therapy (aPDT) has been an alternative treatment. This technique uses a photosensitizer that is activated by light with a specific wavelength producing reactive species, leading to the death of pathogenic microorganisms. In this study, bacteriochlorophyll derivatives such as bacteriochlorin metoxi (Bchl-M) and bacteriochlorin trizma (Bchl-T) obtained from purple bacterium (Rhodopseudomonas faecalis), were evaluated as photosensitizers in the aPDT. Photodynamic inactivation (PDI) of the microorganisms Staphylococcus aureus, Micrococcus luteus, Candida albicans and Pseudomonas aeruginosa was investigated with both bacteriochlorins (Bchl-M and Bchl-T) at different concentrations (1, 15 and 30 µM for S. aureus; 1, 15, 30, 45, 60 and 75 µM for M. luteus; 30, 60, 90, 105, 120 and 150 µM for C. albicans; and 200 µM for P. aeruginosa) and different doses of light (20 and 30 J/cm2 for S. aureus and M. luteus; 30 and 45 J/cm2 for C. albicans; and 45 J/cm2 for P. aeruginosa) to inactivate them. Both photosensitizers showed good activation against S. aureus and for M. luteus, we observed the inactivation of these microorganisms at approximately 3 log, showing to be a good photosensitizers for these microorganisms.


Assuntos
Candida albicans , Luz , Fotoquimioterapia , Fármacos Fotossensibilizantes , Pseudomonas aeruginosa , Staphylococcus aureus , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/química , Candida albicans/efeitos dos fármacos , Candida albicans/efeitos da radiação , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos da radiação , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/efeitos da radiação , Fotoquimioterapia/métodos , Porfirinas/farmacologia , Porfirinas/química , Viabilidade Microbiana/efeitos dos fármacos , Viabilidade Microbiana/efeitos da radiação , Micrococcus luteus/efeitos dos fármacos , Micrococcus luteus/efeitos da radiação , Bactérias/efeitos dos fármacos , Bactérias/efeitos da radiação
3.
Arch Microbiol ; 206(1): 26, 2023 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-38108901

RESUMO

Due to their non-toxic and non-carcinogenic nature, biopigments have a phenomenal benefit over synthetic pigments, making them a desirable source for human utilization and a potential alternative to traditional synthetic pigments that are hazardous to the environment and public health. Endosymbiotic interactions between mangrove plants and bacteria could provide an alternate source for the synthesis of unique compounds with potent biomedical applications. Pigmented endophytic bacteria were screened from the explants of Avicennia marina, a mangrove plant, and identified as Micrococcus luteus by molecular characterization. The intracellular pigment was successfully extracted using the sonication-assisted solvent extraction method, and screening factors impacting the pigmentation bioprocess were determined using a one-factor-at-a-time approach. The endophyte produced yellow pigment in the liquid medium, with the maximum growth and pigment production recorded in nutrient broth at 37 â„ƒ and pH 7 after 96 h of incubation, while the maximum accumulation of pigment was observed in the media supplemented with glucose and tryptone as carbon and nitrogen sources, respectively. The extracted crude pigment was further characterized by ultraviolet, followed by Fourier transform infrared spectroscopy and gas chromatography-mass spectrometry. The obtained crude pigment has been evaluated for its antioxidant and anticancer activity by various assays, such as DPPH radical scavenging activity, FRAP assay, superoxide anion and nitric oxide radical scavenging, metal chelating activity, phosphomolybdenum assay, and MTT assay, respectively, at varying concentrations. The results of our study revealed that the yellow pigment produced by the endophyte showed significant dose-dependent antioxidant and anticancer activity.


Assuntos
Antioxidantes , Avicennia , Humanos , Micrococcus luteus , Nutrientes , Carbono , Endófitos
4.
Exp Mol Med ; 55(1): 196-204, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36639716

RESUMO

Bacterial extracellular vesicles (EVs) have been shown to regulate various pulmonary diseases, but their functions in asthma remain uncertain. To demonstrate the clinical significance of Micrococcus luteus-derived EVs (MlEVs) in asthma, we enrolled 45 asthmatic patients (20 patients with neutrophilic asthma [NA], 25 patients with eosinophilic asthma [EA]) and 40 healthy controls (HCs). When the prevalence of IgG1 and IgG4 specific to MlEVs was evaluated in serum by ELISA, lower levels of MlEV-specific IgG4 (but not IgG1) were noted in asthmatic patients than in HCs. Among asthmatic patients, significantly lower levels of MIEV-specific IgG4 were noted in patients with NA than in those with EA. Moreover, there was a positive correlation between serum MlEV-specific IgG4 levels and FEV1 (%) values. In asthmatic C57BL/6 mice, MlEVs significantly attenuated neutrophilic airway inflammation by reducing the production of IL-1ß and IL-17 in bronchoalveolar lavage fluid as well as the number of group 3 innate lymphoid cells (ILC3s) in lung tissues. To clarify the functional mechanism of MlEVs in NA, the effect of MlEVs on airway epithelial cells (AECs) and immune cells was investigated ex vivo. According to microarray analysis, MlEVs upregulated hsa-miR-4517 expression in AECs. Moreover, this miRNA could suppress IL-1ß production by monocytes, resulting in the inhibition of ILC3 activation and neutrophil recruitment. These findings suggest that MlEVs could be a novel therapeutic agent for managing unresolved NA by regulating miRNA expression in AECs.


Assuntos
Asma , Vesículas Extracelulares , MicroRNAs , Camundongos , Animais , MicroRNAs/metabolismo , Micrococcus luteus/genética , Micrococcus luteus/metabolismo , Imunidade Inata , Camundongos Endogâmicos C57BL , Linfócitos/metabolismo , Líquido da Lavagem Broncoalveolar , Células Epiteliais/metabolismo , Vesículas Extracelulares/metabolismo , Modelos Animais de Doenças
5.
Sci Rep ; 12(1): 18856, 2022 11 07.
Artigo em Inglês | MEDLINE | ID: mdl-36344587

RESUMO

This study investigated the mechanism of membrane damage by protocatechualdehyde (PCA) against Micrococcus luteus and assessed effects of PCA on the sensory and physicochemical properties of pork. The mechanism of PCA inhibition on M. luteus was studied by determining the minimum inhibitory concentration (MIC) based on membrane potential, intracellular ATP concentration, intracellular pH, confocal laser scanning microscopy (CLSM), and field emission gun scanning electron microscopy (FEG-SEM). The results showed that the MIC of PCA against M. luteus was 1.25 mg/mL. Hyperpolarization of the bacterial cell membrane, a decrease in the intracellular ATP concentration, and intracellular pH indicated that PCA damaged the cell membrane of M. luteus. FEG-SEM observation revealed that PCA could cause surface collapse, cell membrane rupture, and content outflow of M. luteus. Additionally, PCA was found to inhibit increases in the total number of colonies, the thiobarbituric acid reactive substances (TBARS) value growth rate, and moisture mobility in raw pork. Additionally, it improved the color and texture of raw pork, all of which effectively prolonged its shelf life. This study will encourage the application of PCA as a natural antibacterial agent in the food industry.


Assuntos
Carne de Porco , Carne Vermelha , Animais , Suínos , Micrococcus luteus , Trifosfato de Adenosina/farmacologia
6.
J Appl Microbiol ; 133(5): 3139-3149, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-35996816

RESUMO

AIM: To reveal the antibacterial mechanism of protocatechuic acid (PCA) against Micrococcus luteus. METHODS AND RESULTS: M. luteus was exposed to PCA, and the antibacterial mechanism was revealed by measuring membrane potential, intracellular ATP and pH levels and transcriptome analysis. PCA induced the membrane potential depolarization of M. luteus, significantly decreased the intracellular ATP and pH levels of M. luteus and disrupted the integrity of the M. luteus cell membrane. Transcriptome analysis showed that PCA induced 782 differentially expressed genes (DEGs) of M. luteus. GO enrichment analysis revealed that the majority of DEGs are involved in pathways of metabolic process, cellular process, biological regulation and transport activity. In addition, PCA inhibited the growth of M. luteus in skimmed milk and extended the shelf life of skimmed milk. CONCLUSION: PCA had good bactericidal activity against M. luteus through the mechanism of cell membrane disruption and metabolic process disorder. SIGNIFICANCE AND IMPACT OF THE STUDY: PCA inhibits the growth of M. luteus in skimmed milk, suggesting that PCA is promising to be used as a novel preservative in food storage.


Assuntos
Perfilação da Expressão Gênica , Micrococcus luteus , Antibacterianos/farmacologia , Trifosfato de Adenosina , Micrococcus
7.
Insect Biochem Mol Biol ; 147: 103775, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35504546

RESUMO

Melanization is a key immune response mediated by serine protease (SP) cascade in insects. Multiple SP pathways exist in different species and it is unclear how conserved these cascades are. The cotton bollworm Helicoverpa armigera is a major worldwide agricultural pest. We reported a conserved melanization pathway in this species, which consists of SP41, cSP1, and cSP6. In this study, we attempted to identify an insect pathogen that elicits the cascade and test whether or not there are other SP cascades in H. armigera. After Micrococcus luteus, Enterobacter cloacae, Beauveria bassiana, or Helicoverpa armigera nucleopolyhedrovirus were injected into larvae, pathogen-induced hemolymph samples were collected for in vitro biochemical assays, which failed to detect proSP41 or procSP1 activation. In contrast, we found that procSP4, a protein proposed to participate in H. armigera melanization, was activated in M. luteus infected hemolymph. We further revealed that cSP8 was a prophenoloxidase (PPO) activating protease downstream of cSP4, and cSP4 was activated by cSP10. The pathway of cSP10-cSP4-cSP8 activated PPO in vitro. Efficiently cleaved procSPH11 and procSPH50 by cSP8 substantially enhanced phenoloxidase activity, suggesting they work together as a cofactor for cSP8 mediated PPO activation. Hemolymph from larvae challenged with M. luteus or its peptidoglycan effectively activated procSP10. Collectively, these results revealed a new PPO activation cascade specifically triggered by the bacterium. In addition, we found that the PPO activation cascades in H. armigera and Manduca sexta are conserved.


Assuntos
Micrococcus luteus , Mariposas , Animais , Catecol Oxidase/metabolismo , Precursores Enzimáticos/metabolismo , Hemolinfa/metabolismo , Proteínas de Insetos/metabolismo , Larva/metabolismo , Micrococcus luteus/metabolismo , Mariposas/metabolismo , Serina Endopeptidases , Serina Proteases/metabolismo
8.
Sci Rep ; 12(1): 2638, 2022 02 16.
Artigo em Inglês | MEDLINE | ID: mdl-35173259

RESUMO

The capture and safe storage of radioactive iodine (129I or 131I) are of a compelling significance in the generation of nuclear energy and waste storage. Because of their physiochemical properties, Porous Organic Polymers (POPs) are considered to be one of the most sought classes of materials for iodine capture and storage. Herein, we report on the preparation and characterization of two triazine-based, nitrogen-rich, porous organic polymers, NRPOP-1 (SABET = 519 m2 g-1) and NRPOP-2 (SABET = 456 m2 g-1), by reacting 1,3,5-triazine-2,4,6-triamine or 1,4-bis-(2,4-diamino-1,3,5-triazine)-benzene with thieno[2,3-b]thiophene-2,5-dicarboxaldehyde, respectively, and their use in the capture of volatile iodine. NRPOP-1 and NRPOP-2 showed a high adsorption capacity of iodine vapor with an uptake of up to 317 wt % at 80 °C and 1 bar and adequate recyclability. The NRPOPs were also capable of removing up to 87% of iodine from 300 mg L-1 iodine-cyclohexane solution. Furthermore, the iodine-loaded polymers, I2@NRPOP-1 and I2@NRPOP-2, displayed good antibacterial activity against Micrococcus luteus (ML), Escherichia coli (EC), and Pseudomonas aeruginosa (PSA). The synergic functionality of these novel polymers makes them promising materials to the environment and public health.


Assuntos
Antibacterianos , Armazenamento de Medicamentos/métodos , Radioisótopos do Iodo , Compostos Orgânicos , Polímeros , Porosidade , Triazinas , Adsorção , Farmacorresistência Bacteriana , Escherichia coli/efeitos dos fármacos , Micrococcus luteus/efeitos dos fármacos , Nitrogênio , Compostos Orgânicos/farmacologia , Polímeros/farmacologia , Triazinas/farmacologia , Volatilização
9.
Sci Rep ; 11(1): 21047, 2021 10 26.
Artigo em Inglês | MEDLINE | ID: mdl-34702916

RESUMO

In recent years' synthesis of metal nanoparticle using plants has been extensively studied and recognized as a non-toxic and efficient method applicable in biomedical field. The aim of this study is to investigate the role of different parts of medical plant Carduus crispus on synthesizing silver nanoparticles and characterize the produced nanoparticle. Our study showed that silver nanoparticles (AgNP) synthesized via whole plant extract exhibited a blue shift in absorption spectra with increased optical density, which correlates to a high yield and small size. Also, the results of zeta potential, X-ray diffraction, photon cross-correlation spectroscopy analysis showed the surface charge of - 54.29 ± 4.96 mV (AgNP-S), - 42.64 ± 3.762 mV (AgNP-F), - 46.02 ± 4.17 mV (AgNP-W), the crystallite size of 36 nm (AgNP-S), 13 nm (AgNP-F), 14 nm (AgNP-W) with face-centered cubic structure and average grain sizes of 145.1 nm, 22.5 nm and 99.6 nm. Another important characteristic, such as elemental composition and constituent capping agent has been determined by energy-dispersive X-ray spectroscopy and Fourier transform infrared. The silver nanoparticles were composed of ~ 80% Ag, ~ 15% K, and ~ 7.5% Ca (or ~ 2.8% P) elements. Moreover, the results of the FTIR measurement suggested that the distinct functional groups present in both AgNP-S and AgNP-F were found in AgNP-W. The atomic force microscopy analysis revealed that AgNP-S, AgNP-F and AgNP-W had sizes of 131 nm, 33 nm and 70 nm respectively. In addition, the biosynthesized silver nanoparticles were evaluated for their cytotoxicity and antibacterial activity. At 17 µg/ml concentration, AgNP-S, AgNP-F and AgNP-W showed very low toxicity on HepG2 cell line but also high antibacterial activity. The silver nanoparticles showed antibacterial activity on both gram-negative bacterium Escherichia coli (5.5 ± 0.2 mm to 6.5 ± 0.3 mm) and gram-positive bacterium Micrococcus luteus (7 ± 0.4 mm to 7.7 ± 0.5 mm). Our study is meaningful as a first observation indicating the possibility of using special plant organs to control the characteristics of nanoparticles.


Assuntos
Antibacterianos , Carduus/química , Escherichia coli/crescimento & desenvolvimento , Nanopartículas Metálicas/química , Micrococcus luteus/crescimento & desenvolvimento , Prata/química , Antibacterianos/síntese química , Antibacterianos/química , Antibacterianos/farmacologia , Células Hep G2 , Humanos
10.
Pol J Microbiol ; 70(3): 321-326, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34584526

RESUMO

Few pieces of research have focused on Micrococcus luteus bloodstream infection (BSI) because of its low incidence; hence data is needed to illustrate this uncommon infection. This study aimed to explore the clinical characteristics of patients with M. luteus BSI. From January 2010 to December 2019, inpatients that met the criteria for M. luteus BSI were included in this study. Data was collected by reviewing electronic records. Ninety-seven patients were enrolled in this study. Sixty-three percent of the patients have a higher neutrophil percentage (NEUT%). The average blood C-reactive protein (CRP) concentration was 5.5 ± 6.4 mg/dl. 48.5% of the patients had malignancy, and 40.2% underwent invasive surgeries. Linezolid was found to have the largest average diameter of the inhibition zone (36 mm), while erythromycin was found to have the smallest average zone diameter (15 mm). However, some M. luteus strains had a potentially broad antimicrobial resistance spectrum. Cephalosporins (59.2%) and quinolones (21.4%) were the most commonly used antibiotics for empirical therapies. In conclusion, M. luteus BSI mainly happens in immunocompromised patients or those with former invasive surgeries or indwelling catheters. M. luteus strains are less responsive to erythromycin. Cephalosporins and quinolones are effective empirical antibiotics for M. luteus BSI; however, vancomycin and teicoplanin should be considered for potentially broadly drug-resistant M. luteus strains.


Assuntos
Infecções por Bactérias Gram-Positivas/patologia , Micrococcus luteus/fisiologia , Sepse/patologia , China , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Sepse/microbiologia , Centros de Atenção Terciária/estatística & dados numéricos
11.
Fish Shellfish Immunol ; 114: 161-170, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33957267

RESUMO

The family of fibrinogen-related proteins (FREPs) is a group of proteins with fibrinogen-like (FBG) domains, which play important roles as pattern recognition receptors (PRRs) in the innate immune responses. In the present study, a fibrinogen-like protein was identified from the oyster Crassostrea gigas (defined as CgFREP1). The open reading frame of CgFREP1 was of 966 bp that encoded a predicted polypeptide of 321 amino acids comprising a signal peptide and a fibrinogen-like domain. The mRNA expression of CgFREP1 was detected in all the examined tissues. The recombinant CgFREP1 (rCgFREP1) displayed binding activities to lipopolysaccharide (LPS), mannose (MAN), as well as Gram-positive bacteria (Micrococcus luteus and Staphylococcus aureus) and Gram-negative bacteria (Vibrio splendidus and Escherichia coli). The rCgFREP1 displayed the agglutinating activity towards M. luteus, V. splendidus and E. coli in the presence of Ca2+. rCgFREP1 was able to enhance the phagocytic activity of haemocytes towards V. splendidus, and exhibited binding activity to the CUB domain of CgMASPL-1. These results suggest that CgFREP1 not only serves as a PRR to recognize and agglutinate different bacteria but also mediates the haemocytes phagocytosis towards V. splendidus.


Assuntos
Crassostrea/microbiologia , Hemócitos/fisiologia , Fagocitose/fisiologia , Proteínas/metabolismo , Vibrio/fisiologia , Animais , Crassostrea/imunologia , Crassostrea/metabolismo , Interações Hospedeiro-Patógeno , Micrococcus luteus/fisiologia , Proteínas/imunologia , Staphylococcus aureus/fisiologia
12.
Dev Comp Immunol ; 122: 104083, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-33930456

RESUMO

Hexokinase (HK) is generally recognized as a crucial enzyme participating in glycolysis. In the present study, a HK (named CgHK) was identified as a potential pattern recognition receptor (PRR) from the Pacific oyster Crassostrea gigas. The open reading frame (ORF) of CgHK was of 1395 bp, encoding a peptide of 464 amino acids with one Hexokinase_1 domain and one Hexokinase_2 domain. The predicted amino acid sequence of CgHK shared 17%-29% similarities with that of other known HKs. The mRNA transcripts of CgHK were constitutively detected in all the examined tissues, with relative high expression level in labial palp and haemocytes. CgHK protein was mainly observed in the cytoplasm of oyster haemocytes. The mRNA expression level of CgHK in haemocytes was significantly up-regulated and peaked at 3 h after Vibrio splendidus (7.64-fold, p < 0.001) and lipopolysaccharide (LPS) (11.86-fold, p < 0.001) stimulations. The recombinant CgHK protein (rCgHK) exhibited Mg2+-dependent adenosine triphosphate (ATP) binding activity in vitro and activity to bind D-(+)-glucose (GLU) and various pathogen-associated molecular pattern (PAMPs) such as LPS and peptidoglycan (PGN) in the absence of Mg2+. It also displayed higher binding activity towards V. splendidus and relatively lower binding activity towards Staphylococcus aureus, Escherichia coli, and Micrococcus luteus. After the mRNA expression of CgHK in haemocytes was knocked down by dsRNA interference, the expression of CgIL17-5 mRNA in haemocytes was considerably down-regulated at 3 h after the stimulation with V. splendidus (0.33-fold, p < 0.001). These results collectively indicated that CgHK was able to recognize various PAMPs and pathogenic bacteria as a PRR apart from being the enzyme to exert ATP binding activity in glycolysis, and activate the anti-bacterial immune response by promoting the expression of pro-inflammatory cytokines CgIL17-5 in oyster haemocytes.


Assuntos
Crassostrea/imunologia , Hexoquinase/metabolismo , Imunidade Inata/imunologia , Receptores de Reconhecimento de Padrão/imunologia , Vibrio/imunologia , Trifosfato de Adenosina/metabolismo , Sequência de Aminoácidos/genética , Animais , Sítios de Ligação/fisiologia , Citocinas/imunologia , Escherichia coli/imunologia , Glucose/metabolismo , Hemócitos/metabolismo , Hexoquinase/genética , Inflamação/imunologia , Lipopolissacarídeos/imunologia , Micrococcus luteus/imunologia , Peptidoglicano/metabolismo , Domínios Proteicos/genética , Interferência de RNA , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Staphylococcus aureus/imunologia
13.
Microb Cell Fact ; 20(1): 30, 2021 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-33531005

RESUMO

BACKGROUND: Protein aggregation is a biological event observed in expression systems in which the recombinant protein is produced under stressful conditions surpassing the homeostasis of the protein quality control system. In addition, protein aggregation is also related to conformational diseases in animals as transmissible prion diseases or non-transmissible neurodegenerative diseases including Alzheimer, Parkinson's disease, amyloidosis and multiple system atrophy among others. At the molecular level, the presence of aggregation-prone domains in protein molecules act as seeding igniters to induce the accumulation of protein molecules in protease-resistant clusters by intermolecular interactions. RESULTS: In this work we have studied the aggregating-prone performance of a small peptide (L6K2) with additional antimicrobial activity and we have elucidated the relevance of the accompanying scaffold protein to enhance the aggregating profile of the fusion protein. Furthermore, we demonstrated that the fusion of L6K2 to highly soluble recombinant proteins directs the protein to inclusion bodies (IBs) in E. coli through stereospecific interactions in the presence of an insoluble protein displaying the same aggregating-prone peptide (APP). CONCLUSIONS: These data suggest that the molecular bases of protein aggregation are related to the net balance of protein aggregation potential and not only to the presence of APPs. This is then presented as a generic platform to generate hybrid protein aggregates in microbial cell factories for biopharmaceutical and biotechnological applications.


Assuntos
Corpos de Inclusão/metabolismo , Peptídeos/metabolismo , Agregados Proteicos , Anti-Infecciosos/farmacologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/metabolismo , Fluorescência , Proteínas de Fluorescência Verde/metabolismo , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Micrococcus luteus/efeitos dos fármacos , Proteínas Recombinantes/metabolismo , Solubilidade , Staphylococcus aureus/efeitos dos fármacos
14.
Vet Res ; 52(1): 32, 2021 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-33632337

RESUMO

Tissue factor pathway inhibitors (TFPI), including TFPI-1 and TFPI-2, are Kunitz-type serine protease inhibitors that mainly inhibit the blood coagulation induced by tissue factors. Previous reports on teleost proved TFPI play important roles in innate immunity. In this study, two TFPI (PoTFPI-1 and PoTFPI-2) molecules from Japanese flounder (Paralichthys olivaceus) were analyzed and characterized for their expression patterns, antibacterial and anticancer activities of the C-terminal derived peptides. Quantitative real time RT-PCR analysis shows that constitutive PoTFPI-1 expression occurred, in increasing order, in the brain, muscle, spleen, gills, head kidney, blood, intestine, heart, and liver; PoTFPI-2 was expressed, in increasing order, in the brain, gills, head kidney, muscle, intestine, spleen, liver, heart, and blood. Under the stimulation of fish pathogens, both PoTFPI-1 and PoTFPI-2 expressions increased significantly in a manner that depended on the pathogens, tissue type, and infection stage. Furthermore, C-terminal peptides TP25 and TP26, derived from PoTFPI-1 and PoTFPI-2, respectively, were synthesized and proved to be active against Micrococcus luteus (for TP25 and TP26) and Staphylococcus aureus (for TP25) via retardation effects on bacterial nucleic acids. In addition, TP25 and TP26 also displayed significant inhibitory effects on human colon cancer cell line HT-29. These results reveal that both PoTFPI-1 and PoTFPI-2 play important roles in host innate immunity. The antibacterial activity and anticancer cells function of TP25 and TP26 will add new insights into the roles of teleost TFPI.


Assuntos
Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Linguados/genética , Linguados/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Sequência de Aminoácidos , Animais , Antibacterianos/farmacologia , Antineoplásicos/farmacologia , Perfilação da Expressão Gênica/veterinária , Glicoproteínas/química , Glicoproteínas/genética , Glicoproteínas/imunologia , Lipoproteínas/química , Lipoproteínas/genética , Lipoproteínas/imunologia , Micrococcus luteus/efeitos dos fármacos , Filogenia , Staphylococcus aureus/efeitos dos fármacos
15.
J Hosp Infect ; 110: 97-102, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33516797

RESUMO

BACKGROUND: Surgical site infections are a global patient safety concern. Due to lack of evidence on contamination, pre-set surgical goods are sometimes disposed of or re-sterilized, thus increasing costs, resource use, and environmental effects. AIM: To investigate time-dependent bacterial air contamination of covered and uncovered sterile goods in the operating room. METHODS: Blood agar plates (N = 1584) were used to detect bacterial air contamination of sterile fields on 48 occasions. Each time, three aerobe and three anaerobe plates were used as baseline to model the preparation time, and 60 (30 aerobe, 30 anaerobe) were used to model the time pending before operation; half of these were covered with sterile drapes and half remained uncovered. Plates were collected after 4, 8, 12, 16, and 24 h. FINDINGS: Mean time before contamination was 2.8 h (95% confidence interval: 2.1-3.4) in the uncovered group and 3.8 h (3.2-4.4) in the covered group (P = 0.005). The uncovered group had 98 colony-forming units (cfu) versus 20 in the covered group (P = 0.0001). Sixteen different micro-organisms were isolated, the most common being Cutibacterium acnes followed by Micrococcus luteus. Of 32 Staphylococcus cfu, 14 were antibiotic resistant, including one multidrug-resistant Staphylococcus epidermidis. CONCLUSION: Protecting sterile fields from bacterial air contamination with sterile covers enhances the durability of sterile goods up to 24 h. Prolonged durability of sterile goods might benefit patient safety, since surgical sterile material could be prepared in advance for acute surgery, thereby enhancing quality of care and reducing both climate impact and costs.


Assuntos
Microbiologia do Ar , Contaminação de Equipamentos , Salas Cirúrgicas , Humanos , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Micrococcus luteus/isolamento & purificação , Propionibacteriaceae/isolamento & purificação , Staphylococcus epidermidis , Infecção da Ferida Cirúrgica/prevenção & controle , Fatores de Tempo
16.
Biochem J ; 478(1): 63-78, 2021 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-33313751

RESUMO

Multidrug resistant (MDR) bacteria have adapted to most clinical antibiotics and are a growing threat to human health. One promising type of candidates for the everlasting demand of new antibiotic compounds constitute antimicrobial peptides (AMPs). These peptides act against different types of microbes by permeabilizing pathogen cell membranes, whereas being harmless to mammalian cells. Contrarily, another class of membrane-active peptides, namely cell-penetrating peptides (CPPs), is known to translocate in eukaryotic cells without substantially affecting the cell membrane. Since CPPs and AMPs share several physicochemical characteristics, we hypothesized if we can rationally direct the activity of a CPP towards antimicrobial activity. Herein, we describe the screening of a synthetic library, based on the CPP sC18, including structure-based design to identify the active residues within a CPP sequence and to discover novel AMPs with high activity. Peptides with increased hydrophobicity were tested against various bacterial strains, and hits were further optimized leading to four generations of peptides, with the last also comprising fluorinated amino acid building blocks. Interestingly, beside strong antibacterial activities, we also detected activity in cancer cells, while non-cancerous cells remained unharmed. The results highlight our new candidates, particularly those from generation 4, as a valuable and promising source for the development of future therapeutics with antibacterial activity and beyond.


Assuntos
Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/química , Antineoplásicos/farmacologia , Bactérias/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Peptídeos Penetradores de Células/química , Peptídeos Catiônicos Antimicrobianos/síntese química , Peptídeos Catiônicos Antimicrobianos/farmacologia , Bacillus subtilis/efeitos dos fármacos , Bacillus subtilis/ultraestrutura , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Peptídeos Penetradores de Células/síntese química , Peptídeos Penetradores de Células/farmacologia , Dicroísmo Circular , Corynebacterium glutamicum/efeitos dos fármacos , Corynebacterium glutamicum/ultraestrutura , Halogenação , Hemólise/efeitos dos fármacos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Concentração Inibidora 50 , Testes de Sensibilidade Microbiana , Micrococcus luteus/efeitos dos fármacos , Microscopia Eletrônica de Varredura , Pseudomonas fluorescens/efeitos dos fármacos , Pseudomonas fluorescens/ultraestrutura
17.
BMC Microbiol ; 20(1): 301, 2020 10 07.
Artigo em Inglês | MEDLINE | ID: mdl-33028252

RESUMO

BACKGROUND: The host-microbial commensalism can shape the innate immune responses in respiratory mucosa and nasal microbiome also modulates front-line immune mechanism in the nasal mucosa. Inhaled allergens encounter the host immune system first in the nasal mucosa, and microbial characteristics of nasal mucus directly impact the mechanisms of initial allergic responses in nasal epithelium. However, the roles of the nasal microbiome in allergic nasal mucosa remain uncertain. We sought to determine the distribution of nasal microbiomes in allergic nasal mucosa and elucidate the interplay between nasal microbiome Staphylococcus species and Th2 cytokines in allergic rhinitis (AR) models. RESULTS: Staphylococcus aureus (AR-SA) and S. epidermidis (AR-SE) were isolated from the nasal mucosa of patients with AR. The influence of nasal microbiome Staphylococcus species on allergic nasal mucosa was also tested with in vitro and in vivo AR models. Pyrosequencing data showed that colonization by S. epidermidis and S. aureus was more dominant in nasal mucus of AR subjects. The mRNA and protein levels of IL-33 and TSLP were significantly higher in AR nasal epithelial (ARNE) cells which were cultured from nasal mucosa of AR subjects, and exposure of ARNE cells to AR-SA reduced IL-33 mRNA and secreted protein levels. Particularly, ovalbumin-driven AR mice inoculated with AR-SA by intranasal delivery exhibited significantly reduced IL-33 in their nasal mucosa. In the context of these results, allergic symptoms and Th2 cytokine levels were significantly downregulated after intranasal inoculation of AR-SA in vivo AR mice. CONCLUSION: Colonization by Staphylococcus species was more dominant in allergic nasal mucosa, and nasal commensal S. aureus from subjects with AR mediates anti-allergic effects by modulating IL-33-dependent Th2 inflammation. The results demonstrate the role of host-bacterial commensalism in shaping human allergic inflammation.


Assuntos
Imunidade Inata , Mucosa Nasal/imunologia , Rinite Alérgica/imunologia , Staphylococcus aureus/imunologia , Staphylococcus epidermidis/imunologia , Simbiose/imunologia , Animais , Corynebacterium/crescimento & desenvolvimento , Citocinas/genética , Citocinas/imunologia , Modelos Animais de Doenças , Enterobacter aerogenes/crescimento & desenvolvimento , Células Epiteliais/imunologia , Células Epiteliais/microbiologia , Feminino , Expressão Gênica , Humanos , Interleucina-33/genética , Interleucina-33/imunologia , Camundongos Endogâmicos BALB C , Micrococcus luteus/crescimento & desenvolvimento , Muco/imunologia , Muco/microbiologia , Mucosa Nasal/microbiologia , Ovalbumina/administração & dosagem , Cultura Primária de Células , RNA Mensageiro/genética , RNA Mensageiro/imunologia , Rinite Alérgica/induzido quimicamente , Rinite Alérgica/microbiologia , Rinite Alérgica/patologia , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus epidermidis/crescimento & desenvolvimento
18.
Biosci Biotechnol Biochem ; 84(8): 1570-1575, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32338185

RESUMO

Chemical screening of culture medium from the soil fungus Stachybotrys sp. resulted in the isolation of the three new phenylspirodrimanes MBJ-0030 (1), MBJ-0031 (2) and MBJ-0032 (3). Their structures were determined by detailed analysis of spectroscopic data. The absolute configurations of 1-3 were determined by modified Mosher's and Marfey's methods. In addition, cytotoxic and antimicrobial evaluations of the compounds were conducted.


Assuntos
Sesquiterpenos Policíclicos/química , Compostos de Espiro/química , Stachybotrys/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Micrococcus luteus/efeitos dos fármacos , Micrococcus luteus/crescimento & desenvolvimento , Sesquiterpenos Policíclicos/isolamento & purificação , Microbiologia do Solo , Compostos de Espiro/isolamento & purificação , Stachybotrys/isolamento & purificação
19.
Biosci. j. (Online) ; 36(2): 606-618, 01-03-2020. tab, ilus
Artigo em Inglês | LILACS | ID: biblio-1146428

RESUMO

The radish (Raphanus sativus L.) is a vegetable of the Brassicaceae family cultivated worldwide and has several medicinal properties. Its biological activities are related to various secondary metabolites present in the species, especially phenolics. Thus, the objectives of this study were the chemical analysis and evaluation of the antioxidant and antimicrobial activities of the dry extract and fractions of the fodder turnip leaves (R. sativus var. oleiferus Metzg.). Samples were analyzed by mass spectrometry and the antioxidant activity was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical method and the reducing power method. Antimicrobial activity was determined by the agar diffusion and microdilution methods. The total phenols were concentrated in the butanol fraction (121.27 mg GAE/g) and the flavonoids were concentrated in the ethyl acetate fraction (98.02 mg EQ/g). The ethyl acetate fraction showed the best antioxidants results, with 83.45% of free radical scavenging and 11.34% of ferric ions reduction. The analysis of antimicrobial activity showed that the dry extract had the highest average zone of inhibition against Bacillus subtilis (18.67 mm). Smaller values of the minimum inhibitory concentration for Micrococcus luteus were, and the ethyl acetate fraction showed a lower minimum inhibitory concentration (0.1 mg/ml) for that microorganism. There was a strong correlation between the antioxidant activity and the content of phenols and flavonoids. The results showed the potential antioxidant and antimicrobial activities of this extract with the ethyl acetate fraction being most promising for further studies.


O rabanete(Raphanus sativus L.) é um vegetal da família Brassicaceae cultivado em todo o mundo e possui diversas propriedades medicinais. Suas atividades biológicas estão relacionadas aos vários metabólitos secundários presentes na espécie, especialmente os compostos fenólicos. Desta forma, os objetivos deste estudo foram realizar análises químicas e avaliar as atividades antioxidante e antimicrobiana do extrato seco e das frações das folhas de R. sativus var. oleiferus Metzg. As amostras foram analisadas em espectrômetro de massas e o potencial antioxidante foi avaliado pelos métodos do radical DPPH (2,2-difenil-1-picrilhidrazila) e do poder redutor. A atividade antimicrobiana foi determinada pelos métodos de difusão em ágar e da microdiluição. Observou-se que os fenóis totais se concentraram na fração butanólica (121,27 mg EAG/g), enquanto que e os teores de flavonoides concentraram-se na fração acetato de etila (98,02 mg EQ/g). A fração acetato de etila apresentou os melhores resultados antioxidantes, com porcentagem de sequestro dos radicais DPPH de 83,45% e com porcentagem de redução dos íons férrico de 11,34%. A análise da atividade antimicrobiana revelou que o extrato seco teve maior média de halos de inibição frente ao Bacillus subtilis(18,67 mm). Os menores valores da concentração inibitória mínima foram para Micrococcus luteus, sendo que a fração acetato de etila demonstrou menor concentração inibitória mínima (0,1 mg/mL) para esse micro-organismo. Houve uma forte correlação entre a atividade antioxidante e o teor de fenóis e de flavonoides. Os resultados demonstraram potenciais ações antioxidante e antimicrobiana do extrato e das frações avaliados, sendo a fração acetato de etila promissora para estudos posteriores.


Assuntos
Raphanus , Anti-Infecciosos , Antioxidantes , Plantas Medicinais , Bacillus subtilis , Micrococcus luteus , Brassicaceae , Compostos Fenólicos , Fenômenos Químicos
20.
Artigo em Inglês | MEDLINE | ID: mdl-31983278

RESUMO

The octahedral Ru(II) complexes containing the 2(2,6-dimethoxypyridine-3-yl)-1H-imidazo(4,5-f)[1, 10]phenanthroline ligand of type [Ru(N-N)2(L)]2+, where N-N = phen (1,10-phenanthroline) (1), bpy (2,2'-bipyridine) (2), and dmb (4,4'-dimethyl-2,2'-bipyridine) (3); L(dmpip) = (2(2,6-dimethoxypyridine-3-yl)1Himidazo(4,5-f)[1, 10]phenanthroline), have been synthesized and characterized by UV-visible absorption, molar conductivity, elemental analysis, mass, IR, and NMR spectroscopic techniques. The physicochemical properties of the Ru(II) complexes were determined by UV-Vis absorption spectroscopy. The DNA binding studies have been explored by UV-visible absorption, fluorescence titrations, and viscosity measurements. The supercoiled pBR322 DNA cleavage efficiency of Ru(II) complexes 1-3 was investigated. The antimicrobial activity of Ru(II) complexes was done against Gram-positive and Gram-negative microorganisms. The in vitro anticancer activities of all the complexes were investigated by cell viability assay, apoptosis, cellular uptake, mitochondrial membrane potential detection, and semi-quantitative PCR on HeLa cells. The result indicates that the synthesized Ru(II) complexes probably interact with DNA through an intercalation mode of binding with complex 1 having slightly stronger DNA binding affinity and anticancer activity than 2 and 3.


Assuntos
Antibacterianos/farmacologia , Antineoplásicos/farmacologia , Complexos de Coordenação/farmacologia , DNA/efeitos dos fármacos , Rutênio/farmacologia , Animais , Antibacterianos/síntese química , Antibacterianos/química , Antineoplásicos/síntese química , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Bacillus megaterium/efeitos dos fármacos , Bacillus megaterium/crescimento & desenvolvimento , Bacillus subtilis/efeitos dos fármacos , Bacillus subtilis/crescimento & desenvolvimento , Sítios de Ligação/efeitos dos fármacos , Bovinos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Complexos de Coordenação/síntese química , Complexos de Coordenação/química , DNA/química , Dano ao DNA , Relação Dose-Resposta a Droga , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Células HeLa , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Micrococcus luteus/efeitos dos fármacos , Micrococcus luteus/crescimento & desenvolvimento , Estrutura Molecular , Plasmídeos/efeitos dos fármacos , Rutênio/química , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Relação Estrutura-Atividade
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