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1.
Sci Rep ; 11(1): 14936, 2021 07 22.
Artigo em Inglês | MEDLINE | ID: mdl-34294789

RESUMO

Methods to spatially induce apoptosis are useful for cancer therapy. To control the induction of apoptosis, methods using light, such as photochemical internalization (PCI), have been developed. We hypothesized that photoinduced delivery of microRNAs (miRNAs) that regulate apoptosis could spatially induce apoptosis. In this study, we identified pre-miR-664a as a novel apoptosis-inducing miRNA via mitochondrial apoptotic pathway. Further, we demonstrated the utility of photoinduced cytosolic dispersion of RNA (PCDR), which is an intracellular RNA delivery method based on PCI. Indeed, apoptosis is spatially regulated by pre-miR-664a and PCDR. In addition, we found that apoptosis induced by pre-miR-664a delivered by PCDR was more rapid than that by lipofection. These results suggest that pre-miR-664a is a nucleic acid drug candidate for cancer therapy and PCDR and pre-miR-664a-based strategies have potential therapeutic uses for diseases affecting various cell types.


Assuntos
MicroRNAs/farmacologia , Neoplasias/genética , Fármacos Fotossensibilizantes/química , Compostos de Quinolínio/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Citosol/química , Células HeLa , Humanos , MicroRNAs/química , Neoplasias/tratamento farmacológico , Ribonucleoproteína Nuclear Pequena U1/química , Ribonucleoproteína Nuclear Pequena U1/genética , Transfecção
2.
Trials ; 22(1): 479, 2021 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-34294123

RESUMO

BACKGROUND: Tonsillectomy is one of the most frequently performed surgeries in children and young adults worldwide. For decades, tonsillectomy was the surgical treatment of choice for recurrent acute tonsillitis. Tonsillotomy was used in some countries as an alternative to tonsillectomy only for the treatment of obstructive sleep apnea in young children. In recent years, an increase of tonsillotomy also to treat recurrent acute tonsillitis can be observed. Therefore, the German Institute for Quality and Efficiency in Health Care (IQWiG) was commissioned by the Federal Joint Committee (G-BA) to investigate whether tonsillotomy offers advantages compared to tonsillectomy. The meta-analysis of the IQWiG including studies until 2016 revealed that the long-term benefits and harms of tonsillotomy compared to tonsillectomy are unclear. Consequently, the G-BA performed a European call for a clinical trial. A consortium of the German Professional Association of ENT-surgeons (BVHNO), the German Society of Oto-Rhino-Laryngology, Head and Neck Surgery (DGHNO-KHC), and the Jena University Hospital were finally selected to perform the TOTO study. METHODS: TOTO is a multicenter, 1:1 two-arm, randomized non-blinded non-inferiority trial. Four hundred fifty-four patients ≥ 3 years of age will be randomly allocated to undergo either tonsillotomy or tonsillectomy as surgical treatment of recurrent acute tonsillitis. All participants will be followed up for a total of 24 months. The primary outcome is the number of sore throat days experienced over the 24-month follow-up. DISCUSSION: TOTO is designed to evaluate the effectiveness and efficiency of tonsillectomy versus tonsillectomy for the management of patients with recurrent acute tonsillitis. Tonsil disease and surgery have a major impact on preschool and school children as well as on economically active young adults, with individual and societal costs through loss of school visits, earnings, and productivity. If tonsillotomy is at least as effective as tonsillectomy but with reduced morbidity, this would reduce costs to the healthcare system and society. TRIAL REGISTRATION: German Clinical Trials Register DRKS00020823 . Registered on 04 September 2020.


Assuntos
Faringite , Apneia Obstrutiva do Sono , Tonsilectomia , Tonsilite , Criança , Pré-Escolar , Humanos , Metanálise como Assunto , Estudos Multicêntricos como Assunto , Compostos de Quinolínio , Ensaios Clínicos Controlados Aleatórios como Assunto , Tiazóis , Tonsilectomia/efeitos adversos , Tonsilite/diagnóstico , Tonsilite/cirurgia , Adulto Jovem
3.
Molecules ; 26(12)2021 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-34205435

RESUMO

The oxazole yellow dye, YOYO-1 (a symmetric homodimer), is a commonly used molecule for staining DNA. We applied the brightness analysis to study the intercalation of YOYO-1 into the DNA. We distinguished two binding modes of the dye to dsDNA: mono-intercalation and bis-intercalation. Bis-intercalation consists of two consecutive mono-intercalation steps, characterised by two distinct equilibrium constants (with the average number of base pair per binding site equals 3.5): K1=3.36±0.43×107M-1 and K2=1.90±0.61×105M-1, respectively. Mono-intercalation dominates at high concentrations of YOYO-1. Bis-intercalation occurs at low concentrations.


Assuntos
Benzoxazóis/química , DNA/química , Substâncias Intercalantes/química , Quinolinas/química , Compostos de Quinolínio/química , Dimerização , Corantes Fluorescentes/química
4.
Molecules ; 26(4)2021 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-33672444

RESUMO

Regioselective synthesis of novel 2H,3H-[1,4]thiazino[2,3,4-ij]quinolin-4-ium derivatives has been developed by annulation reactions of 8-quinolinesulfenyl halides with vinyl chalcogenides (vinyl ethers, divinyl sulfide, divinyl selenide and phenyl vinyl sulfide) and tetravinyl silane. The novel reagent 8-quinolinesulfenyl bromide was used in the annulation reactions. The influence of the substrate structure and the nature of heteroatoms on the direction of the reactions and on product yields has been studied. The opposite regiochemistry was observed in the reactions with vinyl chalcogenides and tetravinyl silane. The obtained condensed heterocycles are novel water-soluble functionalized compounds with promising biological activity.


Assuntos
Compostos de Quinolínio/síntese química , Estrutura Molecular , Compostos de Quinolínio/química , Solubilidade , Estereoisomerismo , Água/química
5.
Opt Express ; 29(5): 7616-7629, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33726259

RESUMO

Anomalous diffusion dynamics in confined nanoenvironments govern the macroscale properties and interactions of many biophysical and material systems. Currently, it is difficult to quantitatively link the nanoscale structure of porous media to anomalous diffusion within them. Fluorescence correlation spectroscopy super-resolution optical fluctuation imaging (fcsSOFI) has been shown to extract nanoscale structure and Brownian diffusion dynamics within gels, liquid crystals, and polymers, but has limitations which hinder its wider application to more diverse, biophysically-relevant datasets. Here, we parallelize the least-squares curve fitting step on a GPU improving computation times by up to a factor of 40, implement anomalous diffusion and two-component Brownian diffusion models, and make fcsSOFI more accessible by packaging it in a user-friendly GUI. We apply fcsSOFI to simulations of the protein fibrinogen diffusing in polyacrylamide of varying matrix densities and super-resolve locations where slower, anomalous diffusion occurs within smaller, confined pores. The improvements to fcsSOFI in speed, scope, and usability will allow for the wider adoption of super-resolution correlation analysis to diverse research topics.


Assuntos
Fibrinogênio/análise , Microscopia de Fluorescência/métodos , Biologia Computacional , Difusão , Corantes Fluorescentes/metabolismo , Compostos de Quinolínio/metabolismo , Análise Espaço-Temporal
6.
J Pharm Sci ; 110(5): 2241-2249, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33549701

RESUMO

The objective of the present work is to prepare and evaluate ionically complexed Quinapyramine sulphate (QS) loaded lipid nanoparticles and its scale up using geometric similarity principle. Docusate sodium (DS), at a molar ratio of 1:2 of QS to DS, was used to prepare hydrophobic Quinapyramine sulphate-Docusate sodium (QS-DS) ionic complex. Based on the difference in total solubility parameter and polarity of QS-DS complex and different lipids, precirol was selected as a lipid for the preparation of lipidic nanoparticles. The particle size, zeta potential, and % entrapment efficiency (%EE) of QS-DS ionic complex loaded solid lipid nanoparticles (QS-DS-SLN) was found to be 250.10 ± 26.04 nm, -27.41 ± 4.18 mV and 81.26 ± 4.67% respectively. FTIR studies confirmed the formation of QS-DS ionic complex. DSC and XRD studies revealed the amorphous nature of QS in QS-DS-SLN. The spherical shape of nanoparticles was confirmed by scanning electron microscopy. QS-DS-SLN showed sustained release of QS for up to 60 h. No significant difference was observed in particle size, zeta potential, and % entrapment efficiency of pilot-scale batch prepared by using rotational speed of 700 rpm. In conclusion, ionic complexation approach can be used to increase % EE of charged drugs into lipid nanoparticles.


Assuntos
Ácido Dioctil Sulfossuccínico , Nanopartículas , Portadores de Fármacos , Lipídeos , Tamanho da Partícula , Compostos de Quinolínio , Sulfatos
7.
J Enzyme Inhib Med Chem ; 36(1): 437-449, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33467931

RESUMO

The pyridinium-2-carbaldoximes with quinolinium carboxamide moiety were designed and synthesised as cholinesterase reactivators. The prepared compounds showed intermediate-to-high inhibition of both cholinesterases when compared to standard oximes. Their reactivation ability was evaluated in vitro on human recombinant acetylcholinesterase (hrAChE) and human recombinant butyrylcholinesterase (hrBChE) inhibited by nerve agent surrogates (NIMP, NEMP, and NEDPA) or paraoxon. In the reactivation screening, one compound was able to reactivate hrAChE inhibited by all used organophosphates and two novel compounds were able to reactivate NIMP/NEMP-hrBChE. The reactivation kinetics revealed compound 11 that proved to be excellent reactivator of paraoxon-hrAChE better to obidoxime and showed increased reactivation of NIMP/NEMP-hrBChE, although worse to obidoxime. The molecular interactions of studied reactivators were further identified by in silico calculations. Molecular modelling results revealed the importance of creation of the pre-reactivation complex that could lead to better reactivation of both cholinesterases together with reducing particular interactions for lower intrinsic inhibition by the oxime.


Assuntos
Inibidores da Colinesterase/farmacologia , Compostos de Piridínio/farmacologia , Compostos de Quinolínio/farmacologia , Acetilcolinesterase/metabolismo , Butirilcolinesterase/metabolismo , Inibidores da Colinesterase/síntese química , Inibidores da Colinesterase/química , Relação Dose-Resposta a Droga , Humanos , Simulação de Acoplamento Molecular , Estrutura Molecular , Compostos de Piridínio/síntese química , Compostos de Piridínio/química , Compostos de Quinolínio/síntese química , Compostos de Quinolínio/química , Proteínas Recombinantes/metabolismo , Relação Estrutura-Atividade
8.
J Ethnopharmacol ; 270: 113848, 2021 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-33485977

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Cinobufacini is extracted from the skins and parotid venom glands of the toad for treating symptoms like swelling and pain in ancient times. Nowadays, cinobifucini injection has also achieved satisfactory therapeutic effects on hepatocellular carcinoma (HCC) in China. AIM OF THE STUDY: Our previous work found that bufothionine, an alkaloid abundant in cinobufacini injection, induced mitochondria-mediated apoptosis. In this work, the underlying effects of bufothionine on autophagy in HCC and its possible dependent pathway were investigated. METHODS: CCK-8 and Hoechst staining assays were performed to verify effects of drugs on proliferation and apoptosis of SMMC7721 cell. H22-tumor-bearing mice model was established by inoculating ascites fluid. HE staining was used to observe pathological changes in liver and tumor tissues. ELISA and Western blot experiments were conducted to investigate IL-6/JAK2/STAT3 signaling pathway. The effects of drugs on expressions of autophagic relative proteins were investigated by Western blot in vitro and in vivo. RESULTS: In vitro, CCK-8 and Hoechst staining assays showed that bufothionine inhibited SMMC7721 cell proliferation and promoted apoptosis at 100 µM. In vivo, bufothionine relieved symptoms of H22-tumor-bearing mice and exerted anti-inflammation activity. ELISA and Western blot demonstrated that bufothionine significantly reduced serum IL-6 concentration, suppressed p-Stat3tyr705, p-Stat3ser727 and Jak2 expressions in tumor tissues and upregulated Atg5, Atg7 and LC3Ⅱ expressions in SMMC7721 cell and H22 tumor. CONCLUSION: This is the first report showing that bufothionine might induce autophagy in HCC by inhibiting JAK2/STAT3 pathway, presenting a possible anti-cancer mechanism of bufothionine in cinobufacini injection.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Autofagia/efeitos dos fármacos , Bufanolídeos/farmacologia , Alcaloides Indólicos/farmacologia , Neoplasias Hepáticas Experimentais/tratamento farmacológico , Neoplasias/patologia , Compostos de Quinolínio/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Antineoplásicos Fitogênicos/uso terapêutico , Apoptose/efeitos dos fármacos , Bufanolídeos/química , Bufanolídeos/uso terapêutico , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Humanos , Alcaloides Indólicos/uso terapêutico , Interleucina-6/antagonistas & inibidores , Janus Quinase 2/antagonistas & inibidores , Janus Quinase 2/metabolismo , Neoplasias Hepáticas Experimentais/metabolismo , Neoplasias Hepáticas Experimentais/patologia , Masculino , Camundongos , Neoplasias/metabolismo , Compostos de Quinolínio/uso terapêutico , Fator de Transcrição STAT3/antagonistas & inibidores , Fator de Transcrição STAT3/metabolismo
9.
J Phys Chem B ; 125(2): 518-527, 2021 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-33426891

RESUMO

Fluorescent DNA-binding dyes are extensively employed as probe and biosensing in biological detection and imaging. Experiments and theoretical calculations of thiazole orange homodimeric (TOTO) dye binding to a single-strand DNA (ssDNA), poly(dG)n (n = 2, 4, 6, 8), reveal that the n = 6 complex shows about 300-fold stronger fluorescence than n = 2, 4 and a slightly stronger one than n = 8 complexes, which is benefited from the length match between TOTO and poly(dG)6. The machine learning, based on molecular dynamics trajectories, indicates that TOTO is featured by the dihedral angle along its backbone and its end-to-end distance, in which the latter one defines the stretch and hairpin structures of TOTO, respectively. The time-dependent density functional theory calculations on the low-lying excited states show that the stretched TOTO with π-π end-stacking binding mode can bring about strong fluorescence with localized π-π* transitions. For the n = 2, 4, and 8 complexes, the linear scaling quantum mechanics calculations indicate that the dominant hairpin TOTO with intercalative binding modes have relatively larger binding energies, leading to fluorescence quenching by intramolecular charge transfer. Our results may provide an insight for modulating the DNA-dye binding modes to tune the degree of charge transfer and designing fluorescent probes for the recognition of specific DNA sequences.


Assuntos
DNA , Substâncias Intercalantes , Fluorescência , Corantes Fluorescentes , Compostos de Quinolínio , Tiazóis
10.
Nucleic Acids Res ; 49(2): e8, 2021 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-33231685

RESUMO

Whole-genome mapping technologies have been developed as a complementary tool to provide scaffolds for genome assembly and structural variation analysis (1,2). We recently introduced a novel DNA labeling strategy based on a CRISPR-Cas9 genome editing system, which can target any 20bp sequences. The labeling strategy is specifically useful in targeting repetitive sequences, and sequences not accessible to other labeling methods. In this report, we present customized mapping strategies that extend the applications of CRISPR-Cas9 DNA labeling. We first design a CRISPR-Cas9 labeling strategy to interrogate and differentiate the single allele differences in NGG protospacer adjacent motifs (PAM sequence). Combined with sequence motif labeling, we can pinpoint the single-base differences in highly conserved sequences. In the second strategy, we design mapping patterns across a genome by selecting sets of specific single-guide RNAs (sgRNAs) for labeling multiple loci of a genomic region or a whole genome. By developing and optimizing a single tube synthesis of multiple sgRNAs, we demonstrate the utility of CRISPR-Cas9 mapping with 162 sgRNAs targeting the 2Mb Haemophilus influenzae chromosome. These CRISPR-Cas9 mapping approaches could be particularly useful for applications in defining long-distance haplotypes and pinpointing the breakpoints in large structural variants in complex genomes and microbial mixtures.


Assuntos
Sistemas CRISPR-Cas , Mapeamento Cromossômico/métodos , Cromossomos Bacterianos/genética , Haemophilus influenzae/genética , RNA Guia/genética , Alelos , Sequência de Bases , Benzoxazóis/análise , Simulação por Computador , Sequência Conservada/genética , RNA Polimerases Dirigidas por DNA , Farmacorresistência Bacteriana/genética , Corantes Fluorescentes/análise , Edição de Genes/métodos , Genoma Bacteriano , Genoma Humano , Haemophilus influenzae/efeitos dos fármacos , Haplótipos/genética , Humanos , Dispositivos Lab-On-A-Chip , Ácido Nalidíxico/farmacologia , Novobiocina/farmacologia , Motivos de Nucleotídeos/genética , Polimorfismo de Nucleotídeo Único , Compostos de Quinolínio/análise , RNA Guia/síntese química , Sequências Repetitivas de Ácido Nucleico/genética , Alinhamento de Sequência , Coloração e Rotulagem/métodos , Proteínas Virais
11.
Chem Commun (Camb) ; 57(4): 480-483, 2021 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-33326520

RESUMO

A novel water-soluble near-infrared fluorescent probe named QX-P with simple synthesis is developed. QX-P has high sensitivity and selectivity to ALP. Moreover, the probe can not only visualize ALP activity in four cell lines, but also real-time image ALP activity during the diagnosis and treatment of diabetes in mice.


Assuntos
Fosfatase Alcalina/metabolismo , Diabetes Mellitus Experimental/enzimologia , Corantes Fluorescentes/química , Animais , Linhagem Celular Tumoral , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/toxicidade , Fluorometria , Humanos , Limite de Detecção , Camundongos , Compostos de Quinolínio/síntese química , Compostos de Quinolínio/química , Compostos de Quinolínio/toxicidade , Espectrometria de Fluorescência , Xantenos/síntese química , Xantenos/química , Xantenos/toxicidade
12.
PLoS One ; 15(12): e0243361, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33275626

RESUMO

We have developed a novel methodology for the delivery of cell-impermeable molecules, based on electrical short-circuiting via a water droplet in dielectric oil. When a cell suspension droplet is placed between a pair of electrodes with an intense DC electric field, droplet bouncing and droplet deformation, which results in an instantaneous short-circuit, can be induced, depending on the electric field strength. We have demonstrated successful transfection of various mammalian cells using the short-circuiting; however, the molecular mechanism remains to be elucidated. In this study, flow cytometric assays were performed with Jurkat cells. An aqueous droplet containing Jurkat cells and plasmids carrying fluorescent proteins was treated with droplet bouncing or short-circuiting. The short-circuiting resulted in sufficient cell viability and fluorescent protein expression after 24 hours' incubation. In contrast, droplet bouncing did not result in successful gene transfection. Transient membrane pore formation was investigated by uptake of a cell-impermeable fluorescence dye YO-PRO-1 and the influx of calcium ions. As a result, short-circuiting increased YO-PRO-1 fluorescence intensity and intracellular calcium ion concentration, but droplet bouncing did not. We also investigated the contribution of endocytosis to the transfection. The pre-treatment of cells with endocytosis inhibitors decreased the efficiency of gene transfection in a concentration-dependent manner. Besides, the use of pH-sensitive dye conjugates indicated the formation of an acidic environment in the endosomes after the short-circuiting. Endocytosis is a possible mechanism for the intracellular delivery of exogenous DNA.


Assuntos
Endocitose , Técnicas de Transferência de Genes , Terapia Genética , Benzoxazóis/química , Benzoxazóis/farmacologia , Cálcio/química , Cálcio/metabolismo , Eletricidade , Humanos , Células Jurkat , Compostos de Quinolínio/química , Compostos de Quinolínio/farmacologia , Água/química , Água/metabolismo
13.
Molecules ; 25(23)2020 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-33276551

RESUMO

Experimental and theoretical studies on thermodynamic properties of quinolinium-based ionic liquids (ILs) based on bis(trifluoromethylsulfonyl)imide anion (namely N-butyl-quinoloinium bis(trifluoromethylsulfonyl)imide, [BQuin][NTf2], N-hexylquinoloinium bis(trifluoromethyl-sulfonyl)imide, [HQuin][NTf2], and N-octylquinoloinium bis(trifluoromethyl-sulfonyl)imide, [OQuin][NTf2]) with aromatic sulfur compounds and heptane, as a model compound of fuel were examined in order to assess the applicability of the studied ionic liquids for desulfurization of fuels. With this aim, the temperature-composition phase diagrams of 13 binary mixtures composed of organic sulfur compounds (thiophene, benzothiophene, or 2-methylthiophene) or heptane and ionic liquid (IL) were investigated at ambient pressure. A dynamic method was used to determine the (solid-liquid) equilibrium phase diagrams in binary systems over a wide composition range and temperature range from T = 255.15 to 365.15 K up to the fusion temperature of ILs. The immiscibility gap with an upper critical solution temperature (UCST) was observed for each binary system under study. The influence of the alkane chain length of the substituent on the IL cation and of the sulfur compounds (the aromaticity of the solvent) was described. The experimental (solid + liquid) phase equilibrium dataset were successfully correlated using the well-known NRTL equation.


Assuntos
Cátions/química , Heptanos/química , Líquidos Iônicos/química , Compostos Orgânicos/química , Transição de Fase , Compostos de Quinolínio/química , Compostos de Enxofre/química , Termodinâmica , Modelos Teóricos , Solubilidade , Solventes/química
14.
Chem Commun (Camb) ; 56(95): 14980-14983, 2020 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-33179655

RESUMO

A dual functional nanoprobe Pd-Q+@PDMS was proposed to simultaneously monitor Cl- and O2, leading to the determination of an average Cl- concentration of 85.7 ± 5.5 mM in lysosomes of HeLa cells. Mimicking ischemic conditions, the cells exhibited a luminescence change corresponding to a decreasing subcellular Cl- concentration.


Assuntos
Cloretos/análise , Complexos de Coordenação/química , Substâncias Luminescentes/química , Lisossomos/química , Nanopartículas/química , Oxigênio/análise , Técnicas Biossensoriais , Cloretos/metabolismo , Glucose/análise , Glucose/metabolismo , Células HeLa , Humanos , Medições Luminescentes , Lisossomos/ultraestrutura , Imagem Óptica , Oxigênio/metabolismo , Paládio/química , Porfirinas/química , Compostos de Quinolínio/química , Dióxido de Silício/química
15.
Sci Rep ; 10(1): 15418, 2020 09 22.
Artigo em Inglês | MEDLINE | ID: mdl-32963303

RESUMO

Streptococcus pneumoniae choline kinase (sChoK) has previously been proposed as a drug target, yet the effectiveness of the first and only known inhibitor of sChoK, HC-3, is in the millimolar range. The aim of this study was thus to further validate sChoK as a potential therapeutic target by discovering more powerful sChoK inhibitors. LDH/PK and colorimetric enzymatic assays revealed two promising sChoK inhibitor leads RSM-932A and MN58b that were discovered with IC50 of 0.5 and 150 µM, respectively, and were shown to be 2-4 magnitudes more potent than the previously discovered inhibitor HC-3. Culture assays showed that the minimum inhibitory concentration (MIC) of RSM-932A and MN58b for S. pneumoniae was 0.4 µM and 10 µM, respectively, and the minimum lethal concentration (MLC) was 1.6 µM and 20 µM, respectively. Western blot monitoring of teichoic acid production revealed differential patterns in response to each inhibitor. In addition, both inhibitors possessed a bacteriostatic mechanism of action, and neither interfered with the autolytic effects of vancomycin. Cells treated with MN58b but not RSM-932A were more sensitive to a phosphate induced autolysis with respect to the untreated cells. SEM studies revealed that MN58b distorted the cell wall, a result consistent with the apparent teichoic acid changes. Two novel and more highly potent putative inhibitors of sChoK, MN58b and RSM-932A, were characterized in this study. However, the effects of sChoK inhibitors can vary at the cellular level. sChoK inhibition is a promising avenue to follow in the development of therapeutics for treatment of S. pneumoniae.


Assuntos
Colina Quinase/metabolismo , Inibidores de Proteínas Quinases/farmacologia , Streptococcus pneumoniae/efeitos dos fármacos , Compostos de Anilina/farmacologia , Autólise/metabolismo , Butanos/farmacologia , Parede Celular/efeitos dos fármacos , Parede Celular/metabolismo , Testes de Sensibilidade Microbiana , Compostos de Piridínio/farmacologia , Compostos de Quinolínio/farmacologia , Streptococcus pneumoniae/metabolismo , Ácidos Teicoicos/metabolismo
16.
Life Sci ; 259: 118180, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32758622

RESUMO

AIMS: Bufothionine had been used for gastric cancer (GC) treatment, and this study managed to uncover the underlying mechanisms. MATERIALS AND METHODS: Cell proliferation was determined by CCK-8 assay and colony formation assay. Flow cytometry (FCM) and TUNEL assay were used to measure cell apoptosis ratio. Intracellular ROS was measured by DCFH-DA probes. qRT-PCR was used to determine miRNAs levels. Western Blot was performed to probe proteins. Dual-luciferase reporter gene system was employed to validate the binding sites of miR-133a-3p and 3'UTR regions of IGF1R mRNA. Immunohistochemistry (IHC) was used to determine the expressions of Ki-67 in mice tumor tissues. KEY FINDINGS: Bufothionine inhibited cell viability, triggered ER stress and promoted ROS production in GC cells, and both ER stress inhibitor Salburinal (Sal) and ROS scavenger (NAC) abrogated Bufothionine induced GC cell death. Besides, miR-133a-3p was upregulated by Bufothionine, and Bufothionine-induced cell death was enhanced by miR-133a-3p overexpression while alleviated by miR-133a-3p knockdown. Furthermore, miR-133a-3p inactivated PI3K/Akt signal pathway by sponging IGF1R, and Bufothionine inhibited insulin-like growth factor 1 receptor (IGF1R) and inactivated PI3K/Akt cascade by upregulating miR-133a-3p. Notably, the promoting effects of overexpressed miR-133a-3p on Bufothionine-induced GC cell death were abrogated by overexpressing IGF1R, and aggravated by the PI3K/Akt cascade inhibitor (LY294002). SIGNIFICANCE: Bufothionine promoted GC cell death by triggering miR-133a-3p/IGF1R/PI3K/Akt axis mediated ER stress and ROS production.


Assuntos
Estresse do Retículo Endoplasmático/efeitos dos fármacos , Estresse do Retículo Endoplasmático/genética , Alcaloides Indólicos/farmacologia , MicroRNAs/genética , Compostos de Quinolínio/farmacologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Neoplasias Gástricas/patologia , Animais , Morte Celular/efeitos dos fármacos , Morte Celular/genética , Proliferação de Células , Cromonas/farmacologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , MicroRNAs/biossíntese , Morfolinas/farmacologia , Proteína Oncogênica v-akt/efeitos dos fármacos , Fosfatidilinositol 3-Quinases/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Receptor IGF Tipo 1/efeitos dos fármacos , Ensaio Tumoral de Célula-Tronco , Regulação para Cima/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
17.
Bioelectrochemistry ; 136: 107624, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32784104

RESUMO

Pulsed radiofrequency (PRF) treatments for chronic pain consist in the delivery of a train of sinusoidal electric bursts to the targeted nerve. Despite numerous clinical evidence of its efficiency, the mechanism of action of PRF remains unclear. Since most of the reported biological effects of PRF can be initiated by a calcium influx into the neurons, we hypothesized that PRF may induce a mild electroporation effect causing a calcium uptake. To test this hypothesis, HEK-293 cells were exposed to PRF bursts and cytosolic calcium and Yo-Pro-1 uptake were monitored. After a single burst, calcium peaks were observed for electric fields above 480 V/cm while the uptake of Yo-pro-1 was insignificant. After a train of 120 bursts, the electric fields required to induce a calcium and Yo-pro-1 uptake decreased to 330 V/cm and 880 V/cm respectively. Calcium peaks were not detected when cells were treated in calcium free media. The temperature increase during the treatments was lower than 5 °C in all cases. Finally, the cell response for different burst frequencies and extracellular media conductivities correlated with the induced transmembrane voltage calculated with a numerical model. Our results support the hypothesis of an electroporation mediated calcium influx.


Assuntos
Cálcio/metabolismo , Dor Crônica/terapia , Eletroporação/métodos , Tratamento por Radiofrequência Pulsada/métodos , Benzoxazóis/metabolismo , Dor Crônica/metabolismo , Citosol/metabolismo , Células HEK293 , Humanos , Técnicas In Vitro , Potenciais da Membrana , Neurônios/metabolismo , Compostos de Quinolínio/metabolismo , Temperatura
18.
Anal Chim Acta ; 1125: 152-161, 2020 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-32674761

RESUMO

Hypoxia, as a condition in which a region of body has low oxygen tension, is closely related to a variety of pathological conditions, and in many diseases local hypoxia occurs that would further increase the severity of diseases. Hence the extent of hypoxia could reflect the related pathological conditions and diseases, and the detection of hypoxia is of great significance. In hypoxia, the elevated level of nitroreductase (NTR) usually occurs, which could serve as a biomarker for hypoxia and thus the related diseases. Herein, an activatable fluorescent probe TPAQS-NO2 based on aggregation-induced emission (AIE) was designed for hypoxia detection via responding to NTR. The probe consists of an electron acceptor quinolinium and an electron donor triphenylamine group. The activated probe shows a large Stokes shift (186 nm). The probe TPAQS-NO2 was successfully used for detecting the early-stage and the advanced-stage tumors via NTR detection in 4T1 tumor-bearing mouse model. Furthermore, the probe TPAQS-NO2 was applied for detecting NTR in the cerebral ischemia (CIS) mouse model. The probe could offer an effective approach for detecting hypoxia-related pathological conditions.


Assuntos
Isquemia Encefálica/diagnóstico por imagem , Corantes Fluorescentes/química , Hipóxia/diagnóstico por imagem , Neoplasias/diagnóstico por imagem , Nitrorredutases/análise , Compostos de Anilina/síntese química , Compostos de Anilina/química , Animais , Isquemia Encefálica/fisiopatologia , Linhagem Celular Tumoral , Corantes Fluorescentes/síntese química , Humanos , Limite de Detecção , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos ICR , Neoplasias/fisiopatologia , Imagem Óptica , Compostos de Quinolínio/síntese química , Compostos de Quinolínio/química
19.
Molecules ; 25(12)2020 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-32630496

RESUMO

Here, we report the syntheses of two pentamethine cyanine dyes containing quinolinium rings and substituted with either hydrogen (3) or bromine (4) at the meso carbon. The electron withdrawing bromine atom stabilizes dye 4 in aqueous buffer, allowing complex formation to occur between the dye and double-helical DNA. UV-visible, CD, and fluorescence spectra recorded at low DNA concentrations suggest that dye 4 initially binds to the DNA as a high-order aggregate. As the ratio of DNA to dye is increased, the aggregate is converted to monomeric and other low-order dye forms that interact with DNA in a non-intercalative fashion. The brominated dye 4 is relatively unreactive in the dark, but, under 707-759 nm illumination, generates hydroxyl radicals that cleave DNA in high yield (pH 7.0, 22 °C). Dye 4 is also taken up by ES2 ovarian carcinoma cells, where it is non-toxic under dark conditions. Upon irradiation of the ES2 cells at 694 nm, the brominated cyanine reduces cell viability from 100 ± 10% to 14 ± 1%. Our results suggest that 2-quinolinium-based carbocyanine dyes equipped with stabilizing electron withdrawing groups may have the potential to serve as sensitizing agents in long-wavelength phototherapeutic applications.


Assuntos
Carbocianinas/química , Clivagem do DNA , DNA de Neoplasias/química , Corantes Fluorescentes/química , Neoplasias Ovarianas/metabolismo , Fotoquímica , Compostos de Quinolínio/química , Apoptose , Proliferação de Células , Feminino , Fluorescência , Humanos , Neoplasias Ovarianas/patologia , Espectroscopia de Luz Próxima ao Infravermelho , Células Tumorais Cultivadas
20.
J Gen Appl Microbiol ; 66(4): 220-227, 2020 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-32507782

RESUMO

The discharge of industrial dyes and their breakdown products are often environmentally harmful. Here, we describe a biodegradation method using Burkholderia multivorans CCA53, which exhibits a capacity to degrade azo dyes, particularly ethyl red. Under the optimized culture conditions, 100 µM ethyl red was degraded more than 99% after incubation for 8 h. Real-time PCR analysis of azoR1 and azoR2, encoding two azoreductases, revealed that transcription level of these genes is enhanced at early phase under the optimized conditions. For a more practical approach, hydrolysates were prepared from eucalyptus or Japanese cedar chips or rice straw, and rice straw hydrolysate was used as the best medium for ethyl red biodegradation. Under those conditions, ethyl red was also degraded with high efficiency (>91%). We have thus constructed a potentially economical method for the biodegradation of ethyl red.


Assuntos
Proteínas de Bactérias/genética , Burkholderia/metabolismo , NADH NADPH Oxirredutases/genética , Compostos de Quinolínio/metabolismo , Biodegradação Ambiental , DNA Bacteriano/genética , Microbiologia Ambiental , Regulação Bacteriana da Expressão Gênica , Nitrorredutases , Filogenia , Reação em Cadeia da Polimerase em Tempo Real
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