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1.
Chemistry ; 30(31): e202400962, 2024 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-38538537

RESUMO

Cost-effective and facile synthetic routes to organic ligands, along with porous materials that exhibit exceptional gas-storage properties, promise significant industrial applications. Here, a two-step synthesis of novel organophosphorus ligands without metal catalysts is reported. These ligands serve as versatile linkers for the construction of metal-organic frameworks (MOFs) incorporating various metal ions, including zinc and copper. One of the zinc-based MOFs demonstrates remarkable gas-storage properties, with a hydrogen (H2) capacity exceeding 2.5 wt% at 77 K and 100 kPa as well as a carbon dioxide (CO2) capacity exceeding 20 wt% at 298 K and 100 kPa. Furthermore, this zinc-based MOF can be synthesized through a solvothermal process on the gram scale that yields high-quality single crystals.

2.
Inorg Chem ; 63(5): 2454-2459, 2024 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-38276883

RESUMO

Single crystals of alkoxy-functionalized hydroxamate/zinc metal-organic frameworks (MOFs) were obtained by fixating the hydroxamate moiety via intramolecular hydrogen bonding. The resulting MOF structures depend on the steric demand of the alkoxy groups, whereby the incorporation of bulky isopropyl groups affords porous hydroxamate/zinc MOFs. The topological structures of the isopropyl-substituted MOFs could be controlled by adding acid.

3.
Chemistry ; 29(25): e202203442, 2023 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-36807627

RESUMO

We have developed highly porous Cu-based metal-organic frameworks (MOFs) using carbazole-type linkers. The novel topological structure of these MOFs was revealed by single-crystal X-ray diffraction analysis. Molecular adsorption/desorption experiments indicated that these MOFs are flexible and change their structure upon adsorption/desorption of organic solvents and gas molecules. These MOFs exhibit unprecedented properties that allow controlling their flexibility by adding a functional group onto the central benzene ring of the organic ligand. For example, the introduction of electron-donating substituents increases the robustness of the resulting MOFs. These MOFs also exhibit flexibility-dependent differences in gas-adsorption and -separation performance. Thus, this study represents the first example of controlling the flexibility of MOFs with the same topological structure via the substituent effect of functional groups introduced into the organic ligand.

4.
Chemistry ; 29(64): e202303417, 2023 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-37916711

RESUMO

Invited for the cover of this issue is the group of Koh Sugamata, Mao Minoura, and co-workers at Rikkyo University. The image depicts triptycene-based metal-organic frameworks with honeycomb structures that collect carbon dioxide and hydrogen, in an analogy to bees collecting honey in their honeycombs. Read the full text of the article at 10.1002/chem.202302080.

5.
Chemistry ; 29(64): e202302080, 2023 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-37589440

RESUMO

A series of metal-organic frameworks (MOFs) based on zinc ions and two triptycene ligands of different size have been synthesized under solvothermal conditions. Structural analyses revealed that they are isostructural 3D-network MOFs. The high porosity and thermal stability of these MOFs can be attributed to the highly rigid triptycene-based ligands. Their BET specific surface areas depend on the size of the triptycene ligands. In contrast to these surface-area data, the H2 and CO2 adsorption of these MOFs is larger for MOFs with small pores. Consequently, we introduced functional groups to the bridge-head position of the triptycene ligands and investigated their effect on the gas-sorption properties. The results unveiled the role of the functional groups in the specific CO2 binding via an induced interaction between adsorbates and the functional groups. Excellent H2 and CO2 properties in these MOFs were achieved in the absence of open metal sites.

6.
J Appl Microbiol ; 132(4): 2957-2967, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-34894031

RESUMO

AIMS: To examine the antifungal action of photocombination treatment with ferulic acid (FA) and ultraviolet-A (UV-A) light (wavelength, 365 nm) by investigating associated changes in cellular functions of Saccharomyces cerevisiae. METHODS AND RESULTS: When pre-incubation of yeast cells with FA was extended from 0.5 to 10 min, its photofungicidal activity increased. Flow cytometry analysis of stained live and dead cells revealed that 10-min UV-A exposure combined with FA (1 mg ml-1 ) induced a ~99.9% decrease in cell viability although maintaining cell membrane integrity when compared with pre-exposure samples. When morphological and biochemical analysis were performed, treated cells exhibited an intact cell surface and oxidative DNA damage similar to control cells. Photocombination treatment induced cellular proteins oxidation, as shown by 2.3-fold increasing in immunostaining levels of ~49-kDa carbonylated proteins compared with pre-irradiation samples. Pyruvate kinase 1 (PK1) was identified by proteomics analysis as a candidate protein whose levels was affected by photocombination treatment. Moreover, intracellular ATP levels decreased following FA treatment both in darkness and with UV-A irradiation, thus suggesting a possible FA-induced delay in cell growth. CONCLUSIONS: FA functions within the cytoplasmic membrane; addition of UV-A exposure induces increased oxidative modifications of cytosolic proteins such as PK1, which functions in ATP generation, without causing detectable genotoxicity, thus triggering inactivation of yeast cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Microbial contamination is a serious problem that diminishes the quality of fruits and vegetables. Combining light exposure with food-grade phenolic acids such as FA is a promising disinfection technology for applications in agriculture and food processing. However, the mode of photofungicidal action of FA with UV-A light remains unclear. This study is the first to elucidate the mechanism using S. cerevisiae. Moreover, proteomics analyses identified a specific cytosolic protein, PK1, which is oxidatively modified by photocombination treatment.


Assuntos
Antifúngicos , Saccharomyces cerevisiae , Ácidos Cumáricos/metabolismo , Ácidos Cumáricos/farmacologia , Saccharomyces cerevisiae/metabolismo , Raios Ultravioleta
7.
Analyst ; 142(3): 472-477, 2017 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-28091627

RESUMO

An immunoassay, which is an indispensable analytical method both in biological research and in medical fields was successfully integrated into a "single-step" by developing a microdevice composed of a graphene oxide (GO)-containing hydrogel and a poly (dimethylsiloxane) (PDMS) microchannel array with a polyethylene glycol (PEG) coating containing a fluorescently-labelled antibody. Here we used 2-hydroxyethylmethacrylate (HEMA) as a monomer that is easily, and homogeneously, mixed with GO to synthesize the hydrogel. The fluorescence quenching and size separation functions were then optimized by controlling the ratios of HEMA and GO. Free fluorescently-labelled antibody was successfully separated from the immunoreaction mixture by the hydrogel network structure, and the fluorescence was subsequently quenched by GO. In comparison to the previously reported immunoassay system using GO, the present system achieved a very high fluorescence resonance energy transfer (FRET) efficiency (∼90%), due to the use of direct adsorption of the fluorescently-labelled antibody to the GO surface; in contrast, the former reported method relied on indirect adsorption of the fluorescently-labelled antibody via immunocomplex formation at the GO surface. Finally, the single-step immunoassay microdevice was made by combining the developed hydrogel and the PDMS microchannel with a coating containing the fluorescently-labelled antibody, and successfully applied for the single-step analysis of IgM levels in diluted human serum by simple introduction of the sample via capillary action.


Assuntos
Anticorpos/química , Grafite , Hidrogéis , Imunoensaio , Fluorescência , Transferência Ressonante de Energia de Fluorescência , Humanos , Imunoglobulina M/sangue , Óxidos , Polietilenoglicóis
8.
Analyst ; 141(11): 3389-94, 2016 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-27127806

RESUMO

A single-step, easy-to-use, and fast capillary-type immunoassay device composed of a polyethylene glycol (PEG) coating containing two kinds of antibody-reagents, including an antibody-graphene oxide conjugate and fluorescently labelled antibody, was developed in this study. The working principle involved the spontaneous dissolution of the PEG coating, diffusion of reagents, and subsequent immunoreaction, triggered by the capillary action-mediated introduction of a sample solution. In a sample solution containing the target antigen, two types of antibody reagents form a sandwich-type antigen-antibody complex and fluorescence quenching takes place via fluorescence resonance energy transfer between the labelled fluorescent molecules and graphene oxide. Antigen concentration can be measured based on the decrease in fluorescence intensity. An antigen concentration-dependent response was obtained for the model target protein sample (human IgG, 0.2-10 µg mL(-1)). The present method can shorten the reaction time to within 1 min (approximately 40 s), while conventional methods using the same reagents require reaction times of approximately 20 min because of the large reaction scale. The proposed method is one of the fastest immunoassays ever reported. Finally, the present device was used to measure human IgG in diluted serum samples to demonstrate that this method can be used for fast medical diagnosis.


Assuntos
Anticorpos Imobilizados/química , Complexo Antígeno-Anticorpo/química , Grafite , Imunoensaio , Imunoconjugados/química , Transferência Ressonante de Energia de Fluorescência , Corantes Fluorescentes , Vidro , Humanos , Imunoglobulina G/sangue , Óxidos , Polietilenoglicóis
9.
Fungal Biol ; 128(2): 1684-1690, 2024 04.
Artigo em Inglês | MEDLINE | ID: mdl-38575241

RESUMO

This study aimed to investigate the effects of ferulic acid (FA), a natural phenolic phytochemical, in combination with light irradiation at three wavelengths (365, 385 and 405 nm) on the concentration and toxicity of deoxynivalenol (DON), a mycotoxin produced by Fusarium graminearum. Moreover, this study examined the influence of the combination treatment on DON production in the cultured fungus. FA activated by light at a peak wavelength of 365 nm exhibited the most effective decrease in DON concentration of the tested wavelengths; a residual DON ratio of 0.23 at 24 h exposure was observed, compared with the initial concentration. The reduction in DON using 365-nm light was dependent on the concentration of FA, with a good correlation (r2 = 0.979) between the rate constants of DON decrease and FA concentration, which was confirmed by a pseudo-first-order kinetics analysis of the photoreaction with different FA concentrations (50-400 mg/L) for 3 h. The viability of HepG2 cells increased by 56.7% following in vitro treatment with a mixture containing the photoproducts obtained after treatment with 20 mg/L DON and 200 mg/L FA under 365-nm irradiation for 6 h. These results suggested that the photoreaction of FA under 365-nm irradiation induces the detoxification of DON through degradation or modification of DON. The antifungal effects of the combination (FA and 365-nm light) on F. graminearum were investigated. Conidia treated with the combination did not show additive or synergistic inhibition of fungal biomass and DON production in 7-day cultivated fungal samples compared with samples after single treatment. However, successive treatment, composed of 90 min irradiation at 365 nm and then treatment with 200 mg/L FA for 90 min in the dark, suppressed fungal growth and DON yield to 70% and 25% of the untreated sample level, respectively. This photo-technology involving the two treatment methods of 365-nm irradiation and FA addition as a food-grade phenolic acid in combination or successively, can aid in developing alternative approaches to eliminate fungal contaminants in the fields of environmental water and agriculture. However, further research is required to explore the underlying mechanisms of DON decontamination and its biosynthesis in F. graminearum.


Assuntos
Ácidos Cumáricos , Fusarium , Micotoxinas , Tricotecenos , Tricotecenos/metabolismo , Micotoxinas/metabolismo , Doenças das Plantas/microbiologia
10.
Microbiol Immunol ; 57(4): 263-72, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23586630

RESUMO

The NfxC-type mutant of Pseudomonas aeruginosa produces the MexEF-OprN efflux pump and down-regulates expression of the quorum-sensing-dependent efflux pump MexAB-OprM and production of virulence factors in the presence of an active transcriptional regulator, MexT. Consequently, these cells are resistant to chloramphenicol and hypersusceptible to ß-lactam antibiotics. An upper negative regulator, MexS, has been assumed to inactivate MexT in wild-type strains, hence shutting down production of the MexEF-OprN pump. This observation was, however, reported in only one clinical strain and not confirmed in well-characterized laboratory strains. Moreover, it is not known whether MexS is involved in the quorum-sensing-dependent regulation of virulence factor production. To assess these issues, a plasmid carrying wild-type mexS was introduced into three NfxC-type mutants from laboratory strains, which carry an impaired mexS and unimpaired mexT. Unexpectedly, all the transformants produced an increased amount of MexEF-OprN proteins. Three clinical NfxC strains were similarly transformed and although MexEF-OprN was undetectable in two of these strains, one produced an increased amount of these proteins, similar to the laboratory strains. These results were interpreted to mean that P. aeruginosa takes two separate routes in MexT-mediated regulation of mexEF-oprN expression: the MexS-bypassed pathway and MexS-mediated pathway. On the other hand, the transformants of both the laboratory and clinically derived NfxC-type cells produced increased amounts of MexAB-OprM and virulence factors, suggesting that production of these proteins occurs via the MexS-mediated pathway.


Assuntos
Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/metabolismo , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana , Plasmídeos/genética , Pseudomonas aeruginosa/efeitos dos fármacos , Percepção de Quorum/genética , Análise de Sequência de DNA , Transformação Bacteriana , Fatores de Virulência/genética , Fatores de Virulência/metabolismo
11.
Theriogenology ; 197: 252-258, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36525864

RESUMO

It is important to prevent contamination inside the incubator as a method of preventing microbial infections during the embryo culture. In the present study, we examined the effects of ultraviolet-C (UV-C) irradiation, used for microorganism inactivation, on embryo development and the growth of bacteria, including Escherichia coli and Staphylococcus aureus, and the fungus Cladosporium cladosporioides. In the embryo irradiation experiment, we examined the effects of the plastic lid of the culture dish, irradiation distances (10, 20, and 25 cm), and different irradiation wavelengths (228 and 260 nm) during embryo culture for 7 days on the development and quality of porcine in vitro-fertilized embryos. None of the embryos cultured in dishes without plastic lids developed into blastocysts after irradiation with 228 nm UV-C. When porcine embryos were cultured in a culture dish with lids, the 228 nm UV-C irradiation decreased blastocyst formation rates of the embryos but not their quality, irrespective of the UV-C irradiation distance. Moreover, irradiation with 260 nm UV-C, even with plastic lids, had more detrimental effects on embryo development than irradiation with 228 nm UV-C. Investigation of the inactivating effects of UV-C irradiation at 228 nm and 260 nm on the growth of the bacteria and fungus showed that 260 nm UV-C reduced the viability to a greater extent than 228 nm UV-C. Moreover, the disinfection efficacy for the bacteria increased when the irradiation duration increased and the distance decreased. In conclusion, porcine embryos can develop into blastocysts without loss of quality even after continuous long-duration irradiation (7 days) with 228 nm UV-C, which can inactivate the growth of bacteria and the tested fungus; however, the development rate of the embryo is reduced.


Assuntos
Blastocisto , Desenvolvimento Embrionário , Animais , Suínos , Desenvolvimento Embrionário/fisiologia , Blastocisto/fisiologia , Embrião de Mamíferos , Fertilização in vitro/veterinária , Escherichia coli , Bactérias , Raios Ultravioleta
12.
Biocontrol Sci ; 17(2): 77-82, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22790843

RESUMO

We previously found that the gemini quaternary salt (gemini-QUAT) containing two pyridinium residues per molecule, 3,3'- (2,7-dioxaoctane) bis (1-decylpyridinium bromide) (3DOBP-4,10) , exerted fungicidal activity against Saccharomyces cerevisiae and caused respiration inhibition and the cytoplasmic leakage of ATP, magnesium, and potassium ions. Here, we investigated how the gemini-QUAT, 3DOBP-4,10, exerts more powerful antimicrobial activity than the mono-QUAT N-cetylpyridinium chloride (CPC) and examined the association between reactive oxygen species (ROS) and the antimicrobial mechanism. Antifungal assays showed that the activity of 3DOBP-4,10 against two yeasts, S. cerevisiae and Candida albicans, was significantly elevated under aerobic conditions, and largely reduced under anaerobic conditions (nitrogen atmosphere) . Adding radical scavengers such as superoxide dismutase, catalase and potassium iodide (KI) also decreased the fungicidal activity of 3DOBP-4,10 but negligibly affected that of CPC. We measured survival under static conditions and found that the rapid fungicidal profile of 3DOBP-4,10 was lost, whereas that of CPC was slightly affected in the presence of KI. Our results suggest that 3DOBP-4,10 exerts powerful antimicrobial activity by penetrating the cell wall and membrane, which then allows oxygen to enter the cells, where it participates in the generation of intracellular ROS. The activity could thus be attributable to a synergic antimicrobial combination of the disruption of organelle membranes by the QUAT and oxidative stress imposed by ROS.


Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Compostos de Piridínio/farmacologia , Compostos de Amônio Quaternário/farmacologia , Espécies Reativas de Oxigênio/farmacologia , Saccharomyces cerevisiae/efeitos dos fármacos , Aerobiose , Anaerobiose , Candida albicans/metabolismo , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Respiração Celular/efeitos dos fármacos , Parede Celular/efeitos dos fármacos , Parede Celular/metabolismo , Sinergismo Farmacológico , Sequestradores de Radicais Livres/farmacologia , Testes de Sensibilidade Microbiana , Estresse Oxidativo/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , Fatores de Tempo
13.
J Photochem Photobiol B ; 229: 112411, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35219030

RESUMO

The aim of this study is to investigate the fungicidal spectrum of six phenolic-cinnamic and -benzoic acid derivatives using four fungi, Aspergillus niger, Cladosporium cladosporioides, Trichophyton mentagrophytes and Candida albicans, in a photocombination system with violet 405-nm light. This is the first study to examine the fungicidal mechanism involving oxidative damage using the conidium of A. niger, as well as an assessment of cellular function and chemical characteristics. The results of the screening assay indicated that ferulic acid (FA) and vanillic acid (VA), which possess 4-hydroxyl and 3-methoxy groups in their phenolic acid structures, produced synergistic activity with 405-nm light irradiation. FA and VA (5.0 mM) significantly decreased the viability of A. niger by 2.4 to 2.6-logs under 90-min irradiation. The synergistic effects were attenuated by the addition of the radical scavenger dimethyl sulfoxide. Generation of reactive oxygen species (ROS), such as hydrogen peroxide and hydroxyl radicals, were confirmed in the phenolic acid solutions tested after irradiation with colorimetric and electron spin resonance analyses. Adsorption of FA and VA to conidia was greater than other tested phenolic acids, and produced 1.55- and 1.85-fold elevation of intracellular ROS levels, as determined using an oxidant-sensitive probe with flow cytometry analysis. However, cell wall or membrane damage was not the main mechanism by which the combination-induced fungal death was mediated. Intracellular ATP was drastically diminished (5% of control levels) following combined treatment with FA and light exposure, even under a condition that produced negligible decreases in viability, thereby resulting in pronounced growth delay. These results suggest that the first stage in the photofungicidal mechanism is oxidative damage to mitochondria or the cellular catabolism system associated with ATP synthesis, which is a result of the photoreaction of phenolic acids adsorbed and internalized by conidia. This photo-technology in combination with food-grade phenolic acids can aid in developing alternative approaches for disinfection of pathogenic and spoilage fungi in the fields of agriculture, food processing and medical care.


Assuntos
Anti-Infecciosos , Hidroxibenzoatos , Anti-Infecciosos/farmacologia , Candida albicans , Hidroxibenzoatos/farmacologia , Espécies Reativas de Oxigênio/metabolismo
14.
Biocontrol Sci ; 27(1): 1-7, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35314556

RESUMO

A kind of citrus fruit with special flavor, Citrus sudachi harvested in Japan, are exported to various countries. However, the Citrus sudachi needs to be sterilized using aqueous solution of sodium hypochlorite because there is a possibility of the adhesion of citrus bacterial canker (CBC) which is not found in Europe. Due to the sterilization with time-consuming work, a more effective decontamination technique is required. A decontamination method using ultraviolet (UV) light irradiation is thus anticipated. Especially, the use of light emitting diodes (LEDs) wi UV light has many advantages in terms of energy consumption, lifetime, and compactness; although an appropriate method is yet to be established. In this study, we evaluate the fundamental effectiveness of UV-LED decontamination on the basis of the bactericidal ability on CBC in petri dishes, using six kinds of UV-LEDs (265, 280, 285, 300, 310, and 365 nm) . For each irradiation, the resultant bactericidal abilities (BAs) were evaluated precisely taking into account the differences in their optical absorptions. In addition, BAs per unit photon number were also estimated, as a fundamental wavelength-dependence of BA. As a result, the effectiveness of UV-LED irradiation with relatively short wavelengths was demonstrated clearly.


Assuntos
Citrus , Raios Ultravioleta , Descontaminação , Frutas , Esterilização
15.
Biocontrol Sci ; 27(1): 9-19, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35314562

RESUMO

The aim of this study was to evaluate bacterial decontamination of boiled young sardine by treatment with violet-blue light followed by cooling storage of the irradiated boiled sardine. Viable cell count in the samples was evaluated after irradiation with four types of violet-blue light-emitting diodes (LEDs; peak wavelength at 405, 412, 421 or 455 nm) and subsequent cooling storage for two days. LED (405 nm) exhibited bactericidal and growth suppression effects. The irradiation gave a 47% bactericidal rate in comparison with no irradiation samples (control) and the two-day storage suppressed the increase in cell counts to 24%, while the rate of increase was 545% for the control. Integrated viability (IV) based on growth delay analysis was estimated after irradiation of four isolates from boiled sardine with 405 nm light. The irradiation caused growth delay against all isolates, resulting in smaller IV values for three isolates compared to those viabilities estimated from colony forming units. Exposure (405 nm) at 432 J/cm2 fluence resulted in a decrease in water content, resulting in an increase in salinity of the samples. This study demonstrated the advantages of light emitting a narrow violet region as a non-thermal disinfection technology in the processing and storage of boiled sardines.


Assuntos
Desinfecção , Contaminação de Medicamentos , Antibacterianos
16.
Biocontrol Sci ; 27(4): 217-222, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36567118

RESUMO

The remote bactericidal effect of TiO2 photocatalyst, i.e., the bactericidal effect away from the photocatalyst, was successfully achieved using a humidified airflow. The TiO2 photocatalyst used was anatase-type TiO2 nanoparticles (NPs) annealed with a low-temperature O2 plasma. For comparison, anatase-type TiO2 NPs annealed in the air were used. The bacteria, Bacillus subtilis, were placed away from the TiO2 NPs. The plasma-assisted-annealed TiO2 NPs significantly inactivated 99% of the bacterial cells in 5 h, whereas the pristine and air-annealed TiO2 NPs inactivated 88-90% of the bacterial cells. The remote bactericidal effect of plasmaassisted-annealed TiO2 NPs would be attributed to a larger amount of H2O2 molecules traveled by the airflow from the TiO2 NPs. The molecules were generated by chemically reacting more photoexcited carriers on the TiO2 surface with H2O and O2 in the airflow. These photoexcited carriers originated from more oxygen-based species adsorbed and more oxygen vacancies introduced on the TiO2 surface by the plasma-assisted-annealing.


Assuntos
Peróxido de Hidrogênio , Nanopartículas , Peróxido de Hidrogênio/farmacologia , Temperatura , Nanopartículas/química , Oxigênio , Antibacterianos/farmacologia
17.
Antimicrob Agents Chemother ; 55(6): 2922-6, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21422216

RESUMO

Serratia marcescens gained resistance to both biocides and antibiotics on expressing the SdeAB efflux pump, following exposure to increasingly higher concentrations of a biocide (H. Maseda et al., Antimicrob. Agents Chemother. 53:5230-5235, 2009). To reveal the regulatory mechanism of sdeAB expression, wild-type cells were subjected to transposon-mediated random mutagenesis, and a mutant with antibiotic resistance, which mimicked the phenotype of the previous biocide-resistant cells, was obtained. The transposon element was found in the chromosomal DNA downstream of the sdeAB operon. Sequencing revealed the presence of an open reading frame (ORF) encoding a protein with 159 amino acid residues that is highly similar to the BadM-type transcriptional repressor, designated sdeS. The level of sdeB::xylE reporter gene expression, undetectable in the wild-type cells, appeared to be fully comparable to that in the biocide-resistant cells. Nucleotide sequencing of the mutant revealed sdeS to have a single G-to-A base substitution at position 269 that converted Trp90 to a stop codon. Introduction of a plasmid-borne intact sdeS into the mutant cells and the biocide-resistant cells resulted in a reduction in sdeB::xylE reporter activity to an undetectable level. These results suggested that SdeS functions as a repressor of the sdeAB operon. It was concluded that the original biocide-resistant cells had an impaired sdeS and, therefore, a derepressed level of the SdeAB efflux pump.


Assuntos
Genes Bacterianos/fisiologia , Óperon , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/genética , Desinfetantes/farmacologia , Farmacorresistência Bacteriana Múltipla , Regulação Bacteriana da Expressão Gênica , Testes de Sensibilidade Microbiana , Norfloxacino/farmacocinética
18.
Front Chem ; 8: 612132, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33409267

RESUMO

In this study, we report an inkjet printing-based method for the immobilization of different reactive analytical reagents on a single microchannel for a single-step and homogeneous solution-based competitive immunoassay. The immunoassay microdevice is composed of a poly(dimethylsiloxane) microchannel that is patterned using inkjet printing by two types of reactive reagents as dissolvable spots, namely, antibody-immobilized graphene oxide and a fluorescently labeled antigen. Since nanoliter-sized droplets of the reagents could be accurately and position-selectively spotted on the microchannel, different reactive reagents were simultaneously immobilized onto the same microchannel, which was difficult to achieve in previously reported capillary-based single-step bioassay devices. In the present study, the positions of the reagent spots and amount of reagent matrix were investigated to demonstrate the stable and reproducible immobilization and a uniform dissolution. Finally, a preliminary application to a single-step immunoassay of C-reactive protein was demonstrated as a proof of concept.

19.
Anticancer Res ; 40(8): 4701-4706, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32727795

RESUMO

BACKGROUND/AIM: Bovine mastitis is caused by the invasion and propagation of pathogenic microorganisms into the udder and mammary gland tissues of cattle. In this study, the therapeutic effect of a low-molecular-weight whey protein (LMW-WP) on bovine mastitis was evaluated. MATERIALS AND METHODS: LMW-WP was orally, intraperitoneally, and vaginally administered to bovine with mastitis. The number of somatic cells in milk was measured 24 h before the administration of LMW-WP. The effect of LMW-WP on cytokine production was measured with a microarray that evaluates the expression of cytokines. RESULTS: In the group that received 1,000 mg intraperitoneally, the somatic cell count was reduced to less than 400,000 at the shipment standard value in three of the four udders, indicating 75% efficacy. The group that received 1,000 mg by vaginal administration showed 67% efficacy. It was confirmed that LMW-WP increased the production of cytokines such as IL-5, IL-6, IL-9, IL-12, MCP-1, and VEGF in mouse macrophage cells, but it did not show any antibacterial activity. CONCLUSION: LMW-WP may be an effective therapeutic agent for bovine mastitis.


Assuntos
Macrófagos/efeitos dos fármacos , Mastite Bovina/tratamento farmacológico , Proteínas do Soro do Leite/farmacologia , Animais , Antibacterianos/farmacologia , Bovinos , Contagem de Células/métodos , Linhagem Celular , Citocinas/metabolismo , Feminino , Glândulas Mamárias Animais/metabolismo , Mastite Bovina/microbiologia , Camundongos , Leite/metabolismo , Células RAW 264.7
20.
Antimicrob Agents Chemother ; 53(12): 5230-5, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19752278

RESUMO

Serratia marcescens is an important opportunistic pathogen in hospitals, where quaternary ammonium compounds are often used for disinfection. The aim of this study is to elucidate the effect of a biocide on the emergence of biocide- and antibiotic-resistant mutants and to characterize the molecular mechanism of biocide resistance in Serratia marcescens. A quaternary ammonium compound-resistant strain, CRes01, was selected by exposing a wild-type strain of S. marcescens to cetylpyridinium chloride. The CRes01 cells exhibited 2- to 16-fold more resistance than the wild-type cells to biocides and antibiotics, including cetylpyridinium chloride, benzalkonium chloride, chlorhexidine gluconate, fluoroquinolones, tetracycline, and chloramphenicol, and showed increased susceptibilities to beta-lactam antibiotics and N-dodecylpyridinium iodide. Mutant cells accumulated lower levels of norfloxacin than the parent cells in an energized state but not in a de-energized state, suggesting that the strain produced a multidrug efflux pump(s). To verify this assumption, we knocked out a putative efflux pump gene, sdeAB, in CRes01 and found that the knockout restored susceptibility to most quaternary ammonium compounds and antibiotics, to which the CRes01 strain showed resistance. On the basis of these and other results, we concluded that S. marcescens gains resistance to both biocides and antibiotics by expressing the SdeAB efflux pump upon exposure to cetylpyridinium chloride.


Assuntos
Proteínas de Bactérias/fisiologia , Cetilpiridínio/farmacologia , Desinfetantes/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Proteínas de Membrana/fisiologia , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/genética , Antibacterianos/metabolismo , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Membrana/genética , Mutação , Norfloxacino/metabolismo , Norfloxacino/farmacologia , Compostos de Amônio Quaternário/farmacologia
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